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Sample records for apical membrane antigen

  1. Identification of a highly antigenic linear B cell epitope within Plasmodium vivax apical membrane antigen 1 (AMA-1.

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    Lilian Lacerda Bueno

    Full Text Available Apical membrane antigen 1 (AMA-1 is considered to be a major candidate antigen for a malaria vaccine. Previous immunoepidemiological studies of naturally acquired immunity to Plasmodium vivax AMA-1 (PvAMA-1 have shown a higher prevalence of specific antibodies to domain II (DII of AMA-1. In the present study, we confirmed that specific antibody responses from naturally infected individuals were highly reactive to both full-length AMA-1 and DII. Also, we demonstrated a strong association between AMA-1 and DII IgG and IgG subclass responses. We analyzed the primary sequence of PvAMA-1 for B cell linear epitopes co-occurring with intrinsically unstructured/disordered regions (IURs. The B cell epitope comprising the amino acid sequence 290-307 of PvAMA-1 (SASDQPTQYEEEMTDYQK, with the highest prediction scores, was identified in domain II and further selected for chemical synthesis and immunological testing. The antigenicity of the synthetic peptide was identified by serological analysis using sera from P. vivax-infected individuals who were knowingly reactive to the PvAMA-1 ectodomain only, domain II only, or reactive to both antigens. Although the synthetic peptide was recognized by all serum samples specific to domain II, serum with reactivity only to the full-length protein presented 58.3% positivity. Moreover, IgG reactivity against PvAMA-1 and domain II after depletion of specific synthetic peptide antibodies was reduced by 18% and 33% (P = 0.0001 for both, respectively. These results suggest that the linear epitope SASDQPTQYEEEMTDYQK is highly antigenic during natural human infections and is an important antigenic region of the domain II of PvAMA-1, suggesting its possible future use in pre-clinical studies.

  2. Identification and characterization of Eimeria tenella apical membrane antigen-1 (AMA1.

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    Lianlian Jiang

    Full Text Available Apical membrane antigen-1 (AMA1 is a micronemal protein of apicomplexan parasites that appears to be essential during the invasion of host cells. In this study, a full-length cDNA of AMA1 was identified from Eimeria tenella (Et using expressed sequence tag and the rapid amplification of cDNA ends technique. EtAMA1 had an open reading frame of 1608 bp encoding a protein of 535 amino acids. Quantitative real-time PCR analysis revealed that EtAMA1 was expressed at higher levels in sporozoites than in the other developmental stages (unsporulated oocysts, sporulated oocysts and second-generation merozoites. The ectodomain sequence was expressed as recombinant EtAMA1 (rEtAMA1 and rabbit polyclonal antibodies raised against the rEtAMA1 recognized a 58-kDa native parasite protein by Western Blotting and had a potent inhibitory effect on parasite invasion, decreasing it by approximately 70%. Immunofluorescence analysis and immunohistochemistry analysis showed EtAMA1 might play an important role in sporozoite invasion and development.

  3. Sequence Analysis of Different Domains of Plasmodium Vivax Apical Membrane Antigen (PvAMA-1 Gene Locus in Iran

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    MR Khoramizade

    2012-02-01

    Full Text Available Background: Plasmodium vivax is responsible for approximately 80 million malaria cases in the world. Apical membrane antigen1 (AMA-1 is a type I integral membrane protein present in all Plasmodium species. AMA-1 interferes in critical steps of invasion of human hepatocytes by sporozoites and red blood cells by merozoites and is one of the most immunodominant antigens for eliciting a protective immune response in human. It is considered as a promising antigen for inclusion in a vaccine against P. vivax. Since more knowledge is needed to lighten the scope of such antigen we compared genetic variation in P. vivax AMA-1from an Iranian isolate with those reported from some of the other malarious countries so far.Methods: P. vivax genomic DNA was extracted from the whole blood of an Iranian patient with patent P. vivax infection. The nucleotide sequence for 446 amino acid (AA residues (42-488 of PvAMA-1 was amplified by PCR and cloned in pUC19 vector for sequencing.Results: Sequence analysis of the antigen showed a high degree of identity (99% with strong homology to the PvAMA-1 gene of P. vivax S3 and SKO814 isolates from India and Korea (Asian isolates respectively, and 96% similarity with P. vivax Sal-1 AMA-1 gene from El Salvador.Conclusions: We cloned and characterized three domains of PvAMA-1 gene from an Iranian patient. Predicted protein sequence of this gene showed some discrepancies in corresponding protein in comparing with similar genes reported from other malarious countries.

  4. Defining the antigenic diversity of Plasmodium falciparum apical membrane antigen 1 and the requirements for a multi-allele vaccine against malaria.

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    Damien R Drew

    Full Text Available Apical Membrane Antigen 1 (AMA1 is a leading malaria vaccine candidate and a target of naturally-acquired human immunity. Plasmodium falciparum AMA1 is polymorphic and in vaccine trials it induces strain-specific protection. This antigenic diversity is a major roadblock to development of AMA1 as a malaria vaccine and understanding how to overcome it is essential. To assess how AMA1 antigenic diversity limits cross-strain growth inhibition, we assembled a panel of 18 different P. falciparum isolates which are broadly representative of global AMA1 sequence diversity. Antibodies raised against four well studied AMA1 alleles (W2Mef, 3D7, HB3 and FVO were tested for growth inhibition of the 18 different P. falciparum isolates in growth inhibition assays (GIA. All antibodies demonstrated substantial cross-inhibitory activity against different isolates and a mixture of the four different AMA1 antibodies inhibited all 18 isolates tested, suggesting significant antigenic overlap between AMA1 alleles and limited antigenic diversity of AMA1. Cross-strain inhibition by antibodies was only moderately and inconsistently correlated with the level of sequence diversity between AMA1 alleles, suggesting that sequence differences are not a strong predictor of antigenic differences or the cross-inhibitory activity of anti-allele antibodies. The importance of the highly polymorphic C1-L region for inhibitory antibodies and potential vaccine escape was assessed by generating novel transgenic P. falciparum lines for testing in GIA. While the polymorphic C1-L epitope was identified as a significant target of some growth-inhibitory antibodies, these antibodies only constituted a minor proportion of the total inhibitory antibody repertoire, suggesting that the antigenic diversity of inhibitory epitopes is limited. Our findings support the concept that a multi-allele AMA1 vaccine would give broad coverage against the diversity of AMA1 alleles and establish new tools to

  5. Sequence diversity and natural selection at domain I of the apical membrane antigen 1 among Indian Plasmodium falciparum populations

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    Kumar Ashwani

    2007-11-01

    Full Text Available Abstract Background The Plasmodium falciparum apical membrane antigen 1 (AMA1 is a leading malaria vaccine candidate antigen. The complete AMA1 protein is comprised of three domains where domain I exhibits high sequence polymorphism and is thus named as the hyper-variable region (HVR. The present study describes the extent of genetic polymorphism and natural selection at domain I of the ama1 gene among Indian P. falciparum isolates. Methods The part of the ama1 gene covering domain I was PCR amplified and sequenced from 157 P. falciparum isolates collected from five different geographical regions of India. Statistical and phylogenetic analyses of the sequences were done using DnaSP ver. 4. 10. 9 and MEGA version 3.0 packages. Results A total of 57 AMA1 haplotypes were observed among 157 isolates sequenced. Forty-six of these 57 haplotypes are being reported here for the first time. The parasites collected from the high malaria transmission areas (Assam, Orissa, and Andaman and Nicobar Islands showed more haplotypes (H and nucleotide diversity π as compared to low malaria transmission areas (Uttar Pradesh and Goa. The comparison of all five Indian P. falciparum subpopulations indicated moderate level of genetic differentiation and limited gene flow (Fixation index ranging from 0.048 to 0.13 between populations. The difference between rates of non-synonymous and synonymous mutations, Tajima's D and McDonald-Kreitman test statistics suggested that the diversity at domain I of the AMA1 antigen is due to positive natural selection. The minimum recombination events were also high indicating the possible role of recombination in generating AMA1 allelic diversity. Conclusion The level of genetic diversity and diversifying selection were higher in Assam, Orissa, and Andaman and Nicobar Islands populations as compared to Uttar Pradesh and Goa. The amounts of gene flow among these populations were moderate. The data reported here will be valuable for the

  6. Genetic polymorphism in domain I of the apical membrane antigen-1 among Plasmodium knowlesi clinical isolates from Peninsular Malaysia.

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    Fong, Mun Yik; Wong, Shen Siang; Silva, Jeremy Ryan De; Lau, Yee Ling

    2015-12-01

    The simian malaria parasite Plasmodium knowlesi is now recognized as a species that can cause human malaria. The first report of large scale human knowlesi malaria was in 2004 in Malaysia Borneo. Since then, hundreds of human knowlesi malaria cases have been reported in Southeast Asia. The present study investigates the genetic polymorphism of P. knowlesi DI domain of the apical membrane antigen-1 (AMA-1), a protein considered as a promising vaccine candidate for malaria. The DI domain of AMA-1 gene of P. knowlesi clinical isolates from Peninsular Malaysia was amplified by PCR, cloned into Escherichia coli, then sequenced and analysed. Ninety-seven DI domain sequences were obtained. Comparison at the nucleotide level against P. knowlesi strain H as reference sequence showed 21 synonymous and 25 nonsynonymous mutations. Nonetheless, nucleotide sequence analysis revealed low genetic diversity of the DI domain, and it was under purifying (negative) selection. At the amino acid level, 26 different haplotypes were identified and 2 were predominant haplotypes (H1, H2) with high frequencies. Phylogenetic analysis revealed that the 26 haplotypes could be clustered into 2 distinct groups (I and II). Members of the groups were basically derived from haplotypes H1 and H2, respectively.

  7. An Extended Surface Loop on Toxoplasma gondii Apical Membrane Antigen 1 (AMA1 Governs Ligand Binding Selectivity.

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    Michelle L Parker

    Full Text Available Apicomplexan parasites are the causative agents of globally prevalent diseases including malaria and toxoplasmosis. These obligate intracellular pathogens have evolved a sophisticated host cell invasion strategy that relies on a parasite-host cell junction anchored by interactions between apical membrane antigens (AMAs on the parasite surface and rhoptry neck 2 (RON2 proteins discharged from the parasite and embedded in the host cell membrane. Key to formation of the AMA1-RON2 complex is displacement of an extended surface loop on AMA1 called the DII loop. While conformational flexibility of the DII loop is required to expose the mature RON2 binding groove, a definitive role of this substructure has not been elucidated. To establish a role of the DII loop in Toxoplasma gondii AMA1, we engineered a form of the protein where the mobile portion of the loop was replaced with a short Gly-Ser linker (TgAMA1ΔDIIloop. Isothermal titration calorimetry measurements with a panel of RON2 peptides revealed an influential role for the DII loop in governing selectivity. Most notably, an Eimeria tenella RON2 (EtRON2 peptide that showed only weak binding to TgAMA1 bound with high affinity to TgAMA1ΔDIIloop. To define the molecular basis for the differential binding, we determined the crystal structure of TgAMA1ΔDIIloop in complex with the EtRON2 peptide. When analyzed in the context of existing AMA1-RON2 structures, spatially distinct anchor points in the AMA1 groove were identified that, when engaged, appear to provide the necessary traction to outcompete the DII loop. Collectively, these data support a model where the AMA1 DII loop serves as a structural gatekeeper to selectively filter out ligands otherwise capable of binding with high affinity in the AMA1 apical groove. These data also highlight the importance of considering the functional implications of the DII loop in the ongoing development of therapeutic intervention strategies targeting the AMA1-RON

  8. High antibody titer against apical membrane antigen-1 is required to protect against malaria in the Aotus model.

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    Sheetij Dutta

    Full Text Available A Plasmodium falciparum 3D7 strain Apical Membrane Antigen-1 (AMA1 vaccine, formulated with AS02(A adjuvant, slowed parasite growth in a recent Phase 1/2a trial, however sterile protection was not observed. We tested this AS02(A, and a Montanide ISA720 (ISA formulation of 3D7 AMA1 in Aotus monkeys. The 3D7 parasite does not invade Aotus erythrocytes, hence two heterologous strains, FCH/4 and FVO, were used for challenge, FCH/4 AMA1 being more homologous to 3D7 than FVO AMA1. Following three vaccinations, the monkeys were challenged with 50,000 FCH/4 or 10,000 FVO parasites. Three of the six animals in the AMA+ISA group were protected against FCH/4 challenge. One monkey did not become parasitemic, another showed only a short period of low level parasitemia that self-cured, and a third animal showed a delay before exhibiting its parasitemic phase. This is the first protection shown in primates with a recombinant P. falciparum AMA1 without formulation in Freund's complete adjuvant. No animals in the AMA+AS02(A group were protected, but this group exhibited a trend towards reduced growth rate. A second group of monkeys vaccinated with AMA+ISA vaccine was not protected against FVO challenge, suggesting strain-specificity of AMA1-based protection. Protection against FCH/4 strain correlated with the quantity of induced antibodies, as the protected animals were the only ones to have in vitro parasite growth inhibitory activity of >70% at 1:10 serum dilution; immuno-fluorescence titers >8,000; ELISA titers against full-length AMA1 >300,000 and ELISA titer against AMA1 domains1+2 >100,000. A negative correlation between log ELISA titer and day 11 cumulative parasitemia (Spearman rank r = -0.780, p value = 0.0001, further confirmed the relationship between antibody titer and protection. High titers of cross-strain inhibitory antibodies against AMA1 are therefore critical to confer solid protection, and the Aotus model can be used to down-select future AMA1

  9. Protein kinase a dependent phosphorylation of apical membrane antigen 1 plays an important role in erythrocyte invasion by the malaria parasite.

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    Kerstin Leykauf

    Full Text Available Apicomplexan parasites are obligate intracellular parasites that infect a variety of hosts, causing significant diseases in livestock and humans. The invasive forms of the parasites invade their host cells by gliding motility, an active process driven by parasite adhesion proteins and molecular motors. A crucial point during host cell invasion is the formation of a ring-shaped area of intimate contact between the parasite and the host known as a tight junction. As the invasive zoite propels itself into the host-cell, the junction moves down the length of the parasite. This process must be tightly regulated and signalling is likely to play a role in this event. One crucial protein for tight-junction formation is the apical membrane antigen 1 (AMA1. Here we have investigated the phosphorylation status of this key player in the invasion process in the human malaria parasite Plasmodium falciparum. We show that the cytoplasmic tail of P. falciparum AMA1 is phosphorylated at serine 610. We provide evidence that the enzyme responsible for serine 610 phosphorylation is the cAMP regulated protein kinase A (PfPKA. Importantly, mutation of AMA1 serine 610 to alanine abrogates phosphorylation of AMA1 in vivo and dramatically impedes invasion. In addition to shedding unexpected new light on AMA1 function, this work represents the first time PKA has been implicated in merozoite invasion.

  10. Multilevel Precision-Based Rational Design of Chemical Inhibitors Targeting the Hydrophobic Cleft of Toxoplasma gondii Apical Membrane Antigen 1 (AMA1).

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    Vetrivel, Umashankar; Muralikumar, Shalini; Mahalakshmi, B; Lily Therese, K; Madhavan, H N; Alameen, Mohamed; Thirumudi, Indhuja

    2016-06-01

    Toxoplasma gondii is an intracellular Apicomplexan parasite and a causative agent of toxoplasmosis in human. It causes encephalitis, uveitis, chorioretinitis, and congenital infection. T. gondii invades the host cell by forming a moving junction (MJ) complex. This complex formation is initiated by intermolecular interactions between the two secretory parasitic proteins-namely, apical membrane antigen 1 (AMA1) and rhoptry neck protein 2 (RON2) and is critically essential for the host invasion process. By this study, we propose two potential leads, NSC95522 and NSC179676 that can efficiently target the AMA1 hydrophobic cleft, which is a hotspot for targeting MJ complex formation. The proposed leads are the result of an exhaustive conformational search-based virtual screen with multilevel precision scoring of the docking affinities. These two compounds surpassed all the precision levels of docking and also the stringent post docking and cumulative molecular dynamics evaluations. Moreover, the backbone flexibility of hotspot residues in the hydrophobic cleft, which has been previously reported to be essential for accommodative binding of RON2 to AMA1, was also highly perturbed by these compounds. Furthermore, binding free energy calculations of these two compounds also revealed a significant affinity to AMA1. Machine learning approaches also predicted these two compounds to possess more relevant activities. Hence, these two leads, NSC95522 and NSC179676, may prove to be potential inhibitors targeting AMA1-RON2 complex formation towards combating toxoplasmosis. PMID:27445648

  11. Multilevel Precision-Based Rational Design of Chemical Inhibitors Targeting the Hydrophobic Cleft of Toxoplasma gondii Apical Membrane Antigen 1 (AMA1).

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    Vetrivel, Umashankar; Muralikumar, Shalini; Mahalakshmi, B; Lily Therese, K; Madhavan, H N; Alameen, Mohamed; Thirumudi, Indhuja

    2016-06-01

    Toxoplasma gondii is an intracellular Apicomplexan parasite and a causative agent of toxoplasmosis in human. It causes encephalitis, uveitis, chorioretinitis, and congenital infection. T. gondii invades the host cell by forming a moving junction (MJ) complex. This complex formation is initiated by intermolecular interactions between the two secretory parasitic proteins-namely, apical membrane antigen 1 (AMA1) and rhoptry neck protein 2 (RON2) and is critically essential for the host invasion process. By this study, we propose two potential leads, NSC95522 and NSC179676 that can efficiently target the AMA1 hydrophobic cleft, which is a hotspot for targeting MJ complex formation. The proposed leads are the result of an exhaustive conformational search-based virtual screen with multilevel precision scoring of the docking affinities. These two compounds surpassed all the precision levels of docking and also the stringent post docking and cumulative molecular dynamics evaluations. Moreover, the backbone flexibility of hotspot residues in the hydrophobic cleft, which has been previously reported to be essential for accommodative binding of RON2 to AMA1, was also highly perturbed by these compounds. Furthermore, binding free energy calculations of these two compounds also revealed a significant affinity to AMA1. Machine learning approaches also predicted these two compounds to possess more relevant activities. Hence, these two leads, NSC95522 and NSC179676, may prove to be potential inhibitors targeting AMA1-RON2 complex formation towards combating toxoplasmosis.

  12. Multilevel Precision-Based Rational Design of Chemical Inhibitors Targeting the Hydrophobic Cleft of Toxoplasma gondii Apical Membrane Antigen 1 (AMA1)

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    Muralikumar, Shalini; Mahalakshmi, B; Lily Therese, K; Madhavan, HN; Alameen, Mohamed; Thirumudi, Indhuja

    2016-01-01

    Toxoplasma gondii is an intracellular Apicomplexan parasite and a causative agent of toxoplasmosis in human. It causes encephalitis, uveitis, chorioretinitis, and congenital infection. T. gondii invades the host cell by forming a moving junction (MJ) complex. This complex formation is initiated by intermolecular interactions between the two secretory parasitic proteins—namely, apical membrane antigen 1 (AMA1) and rhoptry neck protein 2 (RON2) and is critically essential for the host invasion process. By this study, we propose two potential leads, NSC95522 and NSC179676 that can efficiently target the AMA1 hydrophobic cleft, which is a hotspot for targeting MJ complex formation. The proposed leads are the result of an exhaustive conformational search-based virtual screen with multilevel precision scoring of the docking affinities. These two compounds surpassed all the precision levels of docking and also the stringent post docking and cumulative molecular dynamics evaluations. Moreover, the backbone flexibility of hotspot residues in the hydrophobic cleft, which has been previously reported to be essential for accommodative binding of RON2 to AMA1, was also highly perturbed by these compounds. Furthermore, binding free energy calculations of these two compounds also revealed a significant affinity to AMA1. Machine learning approaches also predicted these two compounds to possess more relevant activities. Hence, these two leads, NSC95522 and NSC179676, may prove to be potential inhibitors targeting AMA1-RON2 complex formation towards combating toxoplasmosis. PMID:27445648

  13. Genetic polymorphism and effect of natural selection at domain I of apical membrane antigen-1 (AMA-1) in Plasmodium vivax isolates from Myanmar.

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    Moon, Sung-Ung; Na, Byoung-Kuk; Kang, Jung-Mi; Kim, Jung-Yeon; Cho, Shin-Hyeong; Park, Yun-Kyu; Sohn, Woon-Mok; Lin, Khin; Kim, Tong-Soo

    2010-05-01

    Malaria is endemic or hypoendemic in Myanmar and the country still contributes to the high level of malaria deaths in South-East Asia. Although information on the nature and extent of population diversity within malaria parasites in the country is essential not only for understanding the epidemic situation but also to establish a proper control strategy, very little data is currently available on the extent of genetic polymorphisms of the malaria parasites in Myanmar. In this study, we analyzed the genetic polymorphism and natural selection at domain I of the apical membrane antigen-1 (AMA-1) among Plasmodium vivax Myanmar isolates. A total of 34 distinguishable haplotypes were identified among the 76 isolates sequenced. Comparison with the previously available PvAMA-1 sequences in the GenBank database revealed that 21 of them were new haplotypes that have never been reported till date. The difference between the rate of nonsynonymous (dN) and synonymous (dS) mutations was positive (dN-dS, 0.013+/-0.005), suggesting the domain I is under positive natural selection. The Tajima's D statistics was found to be -0.74652, suggesting that the gene has evolved under population size expansion and/or positive selection. The minimum recombination events were also high, indicating that recombination may occur within the domain I resulting in allelic diversity of PvAMA-1. Our results collectively suggest that PvAMA-1 displays high genetic polymorphism among Myanmar P. vivax isolates with highly diversifying selection at domain I. These results have significant implications in understanding the nature of P. vivax population circulating in Myanmar as well as providing useful information for malaria vaccine development based on this antigen.

  14. Seroreactivity to a Large Panel of Field-Derived Plasmodium falciparum Apical Membrane Antigen 1 and Merozoite Surface Protein 1 Variants Reflects Seasonal and Lifetime Acquired Responses to Malaria

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    Bailey, Jason A.; Pablo, Jozelyn; Niangaly, Amadou; Travassos, Mark A.; Ouattara, Amed; Coulibaly, Drissa; Laurens, Matthew B.; Takala-Harrison, Shannon L.; Lyke, Kirsten E.; Skinner, Jeff; Berry, Andrea A.; Jasinskas, Algis; Nakajima-Sasaki, Rie; Kouriba, Bourema; Thera, Mahamadou A.; Felgner, Philip L.; Doumbo, Ogobara K.; Plowe, Christopher V.

    2015-01-01

    Parasite antigen diversity poses an obstacle to developing an effective malaria vaccine. A protein microarray containing Plasmodium falciparum apical membrane antigen 1 (AMA1, n = 57) and merozoite surface protein 1 19-kD (MSP119, n = 10) variants prevalent at a malaria vaccine testing site in Bandiagara, Mali, was used to assess changes in seroreactivity caused by seasonal and lifetime exposure to malaria. Malian adults had significantly higher magnitude and breadth of seroreactivity to variants of both antigens than did Malian children. Seroreactivity increased over the course of the malaria season in children and adults, but the difference was more dramatic in children. These results help to validate diversity-covering protein microarrays as a promising tool for measuring the breadth of antibody responses to highly variant proteins, and demonstrate the potential of this new tool to help guide the development of malaria vaccines with strain-transcending efficacy. PMID:25294612

  15. The apical plasma membrane of chitin-synthesizing epithelia

    Institute of Scientific and Technical Information of China (English)

    Bernard Moussian

    2013-01-01

    Chitin is the second most abundant polysaccharide on earth.It is produced at the apical side of epidermal,tracheal,fore-,and hindgut epithelial cells in insects as a central component of the protective and supporting extracellular cuticle.Chitin is also an important constituent of the midgut peritrophic matrix that encases the food supporting its digestion and protects the epithelium against invasion by possibly ingested pathogens.The enzyme producing chitin is a glycosyltransferase that resides in the apical plasma membrane forming a pore to extrude the chains of chitin into the extracellular space.The apical plasma membrane is not only a platform for chitin synthases but,probably through its shape and equipment with distinct factors,also plays an important role in orienting and organizing chitin fibers.Here,I review findings on the cellular and molecular constitution of the apical plasma membrane of chitin-producing epithelia mainly focusing on work done in the fruit fly Drosophila melanogaster.

  16. Aquaporin-2: COOH terminus is necessary but not sufficient for routing to the apical membrane.

    NARCIS (Netherlands)

    Deen, P.M.T.; Balkom, B.W.M. van; Savelkoul, P.J.M.; Kamsteeg, E.J.; Raak, M.M.J.P. van; Jennings, M.L.; Muth, T.R.; Rajendran, V.; Caplan, M.J.

    2002-01-01

    Renal regulation of mammalian water homeostasis is mediated by the aquaporin-1 (AQP1) water channel, which is expressed in the apical and basolateral membranes of proximal tubules and descending limbs of Henle, and aquaporin-2 (AQP2), which is redistributed from intracellular vesicles to the apical

  17. Chimaerin suppresses Rac1 activation at the apical membrane to maintain the cyst structure.

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    Shunsuke Yagi

    Full Text Available Epithelial organs are made of a well-polarized monolayer of epithelial cells, and their morphology is maintained strictly for their proper functions. Previously, we showed that Rac1 activation is suppressed at the apical membrane in the mature organoid, and that such spatially biased Rac1 activity is required for the polarity maintenance. Here we identify Chimaerin, a GTPase activating protein for Rac1, as a suppressor of Rac1 activity at the apical membrane. Depletion of Chimaerin causes over-activation of Rac1 at the apical membrane in the presence of hepatocyte growth factor (HGF, followed by luminal cell accumulation. Importantly, Chimaerin depletion did not inhibit extension formation at the basal membrane. These observations suggest that Chimaerin functions as the apical-specific Rac1 GAP to maintain epithelial morphology.

  18. Cholesterol depletion of enterocytes. Effect on the Golgi complex and apical membrane trafficking

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    Hansen, Gert Helge; Niels-Christiansen, L L; Thorsen, Evy;

    2000-01-01

    Intestinal brush border enzymes, including aminopeptidase N and sucrase-isomaltase, are associated with "rafts" (membrane microdomains rich in cholesterol and sphingoglycolipids). To assess the functional role of rafts in the present work, we studied the effect of cholesterol depletion on apical......-associated. Our results implicate the Golgi complex/trans-Golgi network in raft formation and suggest a close relationship between this event and apical membrane trafficking....

  19. Effects of ADH on the apical and basolateral membranes of toad urinary bladder epithelial cells.

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    Donaldson, P J; Leader, J P

    1993-11-01

    Short-circuited urinary bladders from Bufo marinus were supported on their apical surface by an agar mounting method and impaled with microelectrodes via their basolateral membrane. This arrangement provided stable and long-lasting impalements of epithelial cells and yielded reliable membrane potentials and voltage divider ratios (Ra/Rb), where Ra and Rb are apical and basolateral membrane resistances respectively. The membrane potential under short-circuit conditions (Vsc) was -51.4 +/- 2.2 mV (n = 59), while under open-circuit conditions apical membrane potential (Va) and basolateral membrane potential (Vb) were -31.0 +/- 2.4 and 59.5 +/- 2.4 mV, respectively. This yields a "well-shaped" potential profile across the toad urinary bladder, where Va is inversely related to the rate of transport, Isc. Antidiuretic hormone (ADH) produced a hyperpolarisation of Vsc and Vb but had no significant effect on Va. In addition, Ra/Rb was significantly increased by ADH (4.6 +/- 0.5 to 10.2 +/- 3.6). Calculation of individual membrane resistances following the addition of amiloride showed that ADH produced a parallel decrease in Ra and Rb membrane resistance, with the observed increase in Ra/Rb being due to a greater percentage decrease in Rb than in Ra. The ability of ADH to effect parallel changes in apical and basolateral membrane conductance helps to maintain a constant cellular volume despite an increase in transepithelial transport. PMID:8309781

  20. The potassium impermeable apical membrane of insect epithelia: a target for development of safe pesticides

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    William R. Harvey

    1987-01-01

    Full Text Available Columnar cell apical membranes (CCAM in series with goblet cell apical membranes (GCAM form an electroosmotic barrier separating the midgut lumen from epithelial cell cytoplasm. A unique K+ ATPase in GCAM generates three gradients across this barrier. A greater than 180 mV electrical gradient (lumen positive drives amino acid uptake through voltage-dependent K+ symports. A greater than 1000-fold [H+] gradient (lumen alkaline and a greater than 10-fold [K+] gradient (lumen concentrated are adaptations to the high tannin and high K+ content, respectively, in dietary plant material. Agents which act on the apical membrane and disrupt the PD, H+, or K+ gradients are potential insecticides. Insect sensory epithelia and mammalian stria vascularis maintain similar PD and K+ gradients but would not be exposed to ingested anti-apical membrane insecticides. Following the demonstration by Sacchi et al. that Bacillus thuringiensis delta-endotoxin (Bt induces specifically a K+ conductance increase in CCAM vesicles, we find that the K+ channel blocking agent, Ba2+, completely reverses Bt inhibition of the K+-carried short circuit current in the isolated midgut of Manduca sexta. Progress in characterizing the apical membrane includes finding that fluorosulfonylbenzoyladenosine binds specifically to certain GCAM polypeptides and that CCAM vesicles can be mass produced by Ca2+ or Mg2+ precipitation from Manduca sexta midgut.

  1. Isolation of radio-iodinated apical and basal-lateral plasma membranes of toad bladder epithelium.

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    Rodriguez, H J; Edelman, I S

    1979-04-01

    The apical and basal-lateral plasma membranes of toad bladder epithelium were radio-iodinated with the glucose-glucose oxidase-lactoperoxidase system. The covalently bound radio iodine was used as a marker during subcellular fractionation and membrane isolation. Homogenization conditions that ensured rupture of more than 80% of the cells without substantial nuclear damage were defined by Normarski optics. The nuclei were separated by differential centrifugation and the apical and basal-lateral components were resolved by differential and sucrose density gradient centrifugation. The apical components yielded two radioactive bands that were identified as glycocalyx and plasma membrane labeled with 125I. The basal-lateral components yielded a hetero-disperse pattern made up of at least 3 radioactive bands, but the bulk of the activity of ouabain-sensitive ATPase comigrated with only one of these bands. The mitochondia, identified by assays for cytochrome oxidase and NADH cytochrome c reductase activities, were separated from the radio-iodine labeled by centrifugation in sucrose density gradients under isokinetic conditions. The labeled glycocalyx and the slowly migrating components of basal-lateral labeling were separated from the radio-iodinated membranes by centrifugation at 100,000 x g x 1 hr after removal of the mitochrondria by the isokinetic method. The labeled membranes were then subjected to ultracentrifugation in sucrose density gradients under isopycnic conditions; the basal-lateral membranes containing ouabain-sensitive ATP-ase were well resolved from the apical membranes by this method. These results provide a relatively rapid method of attaining partial purification of the apical and basal-lateral plasma membranes of toad bladder epithelium. PMID:222911

  2. Recycling endosomes in apical plasma membrane domain formation and epithelial cell polarity

    NARCIS (Netherlands)

    Golachowska, Magdalena R.; Hoekstra, Dick; van IJzendoorn, Sven C. D.

    2010-01-01

    Recycling endosomes have taken central stage in the intracellular sorting and polarized trafficking of apical and basolateral plasma membrane components. Molecular players in the underlying mechanisms are now emerging, including small GTPases, class V myosins and adaptor proteins. In particular, def

  3. A glycophospholipid membrane anchor acts as an apical targeting signal in polarized epithelial cells

    OpenAIRE

    1989-01-01

    Glycosyl-phosphatidylinositol- (GPI) anchored proteins contain a large extracellular protein domain that is linked to the membrane via a glycosylated form of phosphatidylinositol. We recently reported the polarized apical distribution of all endogenous GPI-anchored proteins in the MDCK cell line (Lisanti, M. P., M. Sargiacomo, L. Graeve, A. R. Saltiel, and E. Rodriguez-Boulan. 1988. Proc. Natl. Acad. Sci. USA. 85:9557-9561). To study the role of this mechanism of membrane anchoring in targeti...

  4. Microbial sphingomyelinase induces RhoA-mediated reorganization of the apical brush border membrane and is protective against invasion.

    Science.gov (United States)

    Saslowsky, David E; Thiagarajah, Jay R; McCormick, Beth A; Lee, Jean C; Lencer, Wayne I

    2016-04-01

    The apical brush border membrane (BBM) of intestinal epithelial cells forms a highly structured and dynamic environmental interface that serves to regulate cellular physiology and block invasion by intestinal microbes and their products. How the BBM dynamically responds to pathogenic and commensal bacterial signals can define intestinal homeostasis and immune function. We previously found that in model intestinal epithelium, the conversion of apical membrane sphingomyelin to ceramide by exogenous bacterial sphingomyelinase (SMase) protected against the endocytosis and toxicity of cholera toxin. Here we elucidate a mechanism of action by showing that SMase induces a dramatic, reversible, RhoA-dependent alteration of the apical cortical F-actin network. Accumulation of apical membrane ceramide is necessary and sufficient to induce the actin phenotype, and this coincides with altered membrane structure and augmented innate immune function as evidenced by resistance to invasion by Salmonella. PMID:26864627

  5. Dependence of intracellular Na+ concentration on apical and basolateral membrane Na+ influx in frog skin

    International Nuclear Information System (INIS)

    An isotopic method was developed to measure the intracellular Na+ content of the transepithelial Na+ transport pool of frog skin. Isolated epithelia (no corium) were labeled with 24Na either asymmetrically, from apical (Aa) or basolateral (Ab) solutions, or symmetrically (Aab). Transport pool Na+ could be identified from the kinetics of washout of 24Na carried out in the presence of 1 mM ouabain, 100 microM amiloride, and 1 mM furosemide that served to trap cold Na+ and 24Na within the transport pool. In control epithelia, Aab averaged 64.1 neq/cm2 (13.9 mM), and maximal inhibition of apical membrane Na+ entry with 100 microM amiloride caused Aab to decrease to 24.3 neq/cm2 (5.3 mM). Ouabain caused Aab to increase markedly to 303 neq/cm2 in 30 min, whereas amiloride inhibition of apical membrane Na+ entry reduced markedly the rate of increase of Aab caused by ouabain. These data, in part, confirmed the existence of an important basolateral membrane permeability to Na+ that was measured in separate studies of the bidirectional 24Na fluxes at the basolateral membranes of the cells. Both sets of data were supportive of the idea that a significant Na+ recycling exists at the basolateral membranes of the cells that contributes to the Na+ load on the pump and Na+ recycling participates in the regulation of the Na+ concentration of the Na+ transport pool of these epithelial cells

  6. Plasma membrane calcium pump (PMCA) isoform 4 is targeted to the apical membrane by the w-splice insert from PMCA2

    OpenAIRE

    Antalffy, Géza; Mauer, Amy S.; Pászty, Katalin; Hegedus, Luca; Padányi, Rita; Enyedi, Ágnes; STREHLER, EMANUEL E.

    2012-01-01

    Local Ca2+ signaling requires proper targeting of the Ca2+ signaling toolkit to specific cellular locales. Different isoforms of the plasma membrane Ca2+ pump (PMCA) are responsible for Ca2+ extrusion at the apical and basolateral membrane of polarized epithelial cells, but the mechanisms and signals for differential targeting of the PMCAs are not well understood. Recent work demonstrated that the alternatively spliced w-insert in PMCA2 directs this pump to the apical membrane. We now show th...

  7. Apical Plasma Membrane Proteins and Endolyn-78 Travel through a Subapical Compartment in Polarized WIF-B Hepatocytes

    OpenAIRE

    Ihrke, Gudrun; Martin, Greg V.; Shanks, Michael R.; Schrader, Michael; Schroer, Trina A.; Hubbard, Ann L.

    1998-01-01

    We studied basolateral-to-apical transcytosis of three classes of apical plasma membrane (PM) proteins in polarized hepatic WIF-B cells and then compared it to the endocytic trafficking of basolaterally recycling membrane proteins. We used antibodies to label the basolateral cohort of proteins at the surface of living cells and then followed their trafficking at 37°C by indirect immunofluorescence. The apical PM proteins aminopeptidase N, 5′nucleotidase, and the polymeric IgA receptor were ef...

  8. Properties of glycolipid-enriched membrane rafts in antigen presentation.

    Science.gov (United States)

    Rodgers, William; Smith, Kenneth

    2005-01-01

    Presentation of antigen to T cells represents one of the central events in the engagement of the immune system toward the defense of the host against pathogens. Accordingly, understanding the mechanisms by which antigen presentation occurs is critical toward our understanding the properties of host defense against foreign antigen, as well as insight into other features of the immune system, such as autoimmune disease. The entire antigen-presentation event is complex, and many features of it remain poorly understood. However, recent studies have provided evidence showing that glycolipid-enriched membrane rafts are important for efficient antigen presentation; the studies suggest that one such function of rafts is trafficking of antigen-MHC II complexes to the presentation site on the surface of the antigen-presenting cell. Here, we present a critical discussion of rafts and their proposed functions in antigen presentation. Emerging topics of rafts and antigen presentation that warrant further investigation are also highlighted.

  9. Cystic fibrosis: model of pathogenesis based on the apical membrane potential

    Directory of Open Access Journals (Sweden)

    Beatrica Kurbel

    2015-02-01

    Full Text Available A simple model of cystic fibrosis (CF is proposed, based on the apical membrane (ApM potential. The ApM of epithelial cells is highly permeable to sodium and activation of CFTRs makes it permeable to chloride. Calculated ApM potentials of cells with activated cystic fibrosis transmembrane conductance regulators (CFTRs are between the sodium and chloride Nernst values and thus allow rapid absorption of both ions in exocrine glands. In CF patients the potential is near the sodium Nernst value and thus more salt is left in the ducts. Simulation predicts that the sodium driving force increases more than 3.5 times if the ApM permeability for Cl- increases from 5-94% of the sodium permeability. In pancreatic ductal cells basolateral sodium bicarbonate cotransporters (pNBC1 allows influx of bicarbonates with sodium. Bicarbonates are exchanged for intraductal chloride by anion exchanger 1 (AE1 in the ApM. Activated CFTRs let some chloride to leak back to ducts, followed by water that dilutes ductal proteins. Replenished intraductal chloride allows more bicarbonate secretion. In CF patients, pancreatic water and bicarbonate secretion is limited by the intraductal chloride pool.

  10. Characterization of plant plasma membrane antigens: [Annual] progress report

    International Nuclear Information System (INIS)

    Protoplast plasma membranes were used to raise antibodies in mice to cell surface antigens. Monoclonal antibodies were selected from those produced and used for indirect immunofluorescence microscopic analysis of N. tabacum cells. In parallel studies cDNA expression libraries were prepared. (DT)

  11. Herpesvirus glycoproteins undergo multiple antigenic changes before membrane fusion.

    Directory of Open Access Journals (Sweden)

    Daniel L Glauser

    Full Text Available Herpesvirus entry is a complicated process involving multiple virion glycoproteins and culminating in membrane fusion. Glycoprotein conformation changes are likely to play key roles. Studies of recombinant glycoproteins have revealed some structural features of the virion fusion machinery. However, how the virion glycoproteins change during infection remains unclear. Here using conformation-specific monoclonal antibodies we show in situ that each component of the Murid Herpesvirus-4 (MuHV-4 entry machinery--gB, gH/gL and gp150--changes in antigenicity before tegument protein release begins. Further changes then occurred upon actual membrane fusion. Thus virions revealed their final fusogenic form only in late endosomes. The substantial antigenic differences between this form and that of extracellular virions suggested that antibodies have only a limited opportunity to block virion membrane fusion.

  12. Small GTPases promote actin coat formation on microsporidian pathogens traversing the apical membrane of Caenorhabditis elegans intestinal cells.

    Science.gov (United States)

    Szumowski, Suzannah C; Estes, Kathleen A; Popovich, John J; Botts, Michael R; Sek, Grace; Troemel, Emily R

    2016-01-01

    Many intracellular pathogens co-opt actin in host cells, but little is known about these interactions in vivo. We study the in vivo trafficking and exit of the microsporidian Nematocida parisii, which is an intracellular pathogen that infects intestinal cells of the nematode Caenorhabditis elegans. We recently demonstrated that N. parisii uses directional exocytosis to escape out of intestinal cells into the intestinal tract. Here, we show that an intestinal-specific isoform of C. elegans actin called ACT-5 forms coats around membrane compartments that contain single exocytosing spores, and that these coats appear to form after fusion with the apical membrane. We performed a genetic screen for host factors required for actin coat formation and identified small GTPases important for this process. Through analysis of animals defective in these factors, we found that actin coats are not required for pathogen exit although they may boost exocytic output. Later during infection, we find that ACT-5 also forms coats around membrane-bound vesicles that contain multiple spores. These vesicles are likely formed by clathrin-dependent compensatory endocytosis to retrieve membrane material that has been trafficked to the apical membrane as part of the exocytosis process. These findings provide insight into microsporidia interaction with host cells, and provide novel in vivo examples of the manner in which intracellular pathogens co-opt host actin during their life cycle. PMID:26147591

  13. Intravacuolar Membranes Regulate CD8 T Cell Recognition of Membrane-Bound Toxoplasma gondii Protective Antigen.

    Science.gov (United States)

    Lopez, Jodie; Bittame, Amina; Massera, Céline; Vasseur, Virginie; Effantin, Grégory; Valat, Anne; Buaillon, Célia; Allart, Sophie; Fox, Barbara A; Rommereim, Leah M; Bzik, David J; Schoehn, Guy; Weissenhorn, Winfried; Dubremetz, Jean-François; Gagnon, Jean; Mercier, Corinne; Cesbron-Delauw, Marie-France; Blanchard, Nicolas

    2015-12-15

    Apicomplexa parasites such as Toxoplasma gondii target effectors to and across the boundary of their parasitophorous vacuole (PV), resulting in host cell subversion and potential presentation by MHC class I molecules for CD8 T cell recognition. The host-parasite interface comprises the PV limiting membrane and a highly curved, membranous intravacuolar network (IVN) of uncertain function. Here, using a cell-free minimal system, we dissect how membrane tubules are shaped by the parasite effectors GRA2 and GRA6. We show that membrane association regulates access of the GRA6 protective antigen to the MHC I pathway in infected cells. Although insertion of GRA6 in the PV membrane is key for immunogenicity, association of GRA6 with the IVN limits presentation and curtails GRA6-specific CD8 responses in mice. Thus, membrane deformations of the PV regulate access of antigens to the MHC class I pathway, and the IVN may play a role in immune modulation. PMID:26628378

  14. Membrane Insertion by Anthrax Protective Antigen in Cultured Cells†

    OpenAIRE

    Qa'Dan, Maen; Christensen, Kenneth A; Zhang, Lei; Roberts, Thomas M.; Collier, R. John

    2005-01-01

    The enzymatic moieties of anthrax toxin enter the cytosol of mammalian cells via a pore in the endosomal membrane formed by the protective antigen (PA) moiety. Pore formation involves an acidic pH-induced conformational rearrangement of a heptameric precursor (the prepore), in which the seven 2β2-2β3 loops interact to generate a 14-strand transmembrane β-barrel. To investigate this model in vivo, we labeled PA with the fluorophore 7-nitrobenz-2-oxa-1,3-diazole (NBD) at cysteine residues intro...

  15. Intravacuolar Membranes Regulate CD8 T Cell Recognition of Membrane-Bound Toxoplasma gondii Protective Antigen

    Directory of Open Access Journals (Sweden)

    Jodie Lopez

    2015-12-01

    Full Text Available Apicomplexa parasites such as Toxoplasma gondii target effectors to and across the boundary of their parasitophorous vacuole (PV, resulting in host cell subversion and potential presentation by MHC class I molecules for CD8 T cell recognition. The host-parasite interface comprises the PV limiting membrane and a highly curved, membranous intravacuolar network (IVN of uncertain function. Here, using a cell-free minimal system, we dissect how membrane tubules are shaped by the parasite effectors GRA2 and GRA6. We show that membrane association regulates access of the GRA6 protective antigen to the MHC I pathway in infected cells. Although insertion of GRA6 in the PV membrane is key for immunogenicity, association of GRA6 with the IVN limits presentation and curtails GRA6-specific CD8 responses in mice. Thus, membrane deformations of the PV regulate access of antigens to the MHC class I pathway, and the IVN may play a role in immune modulation.

  16. Endocytosis Is Crucial for Cell Polarity and Apical Membrane Recycling in the Filamentous Fungus Aspergillus oryzae▿

    OpenAIRE

    Higuchi, Yujiro; Shoji, Jun-ya; Arioka, Manabu; Kitamoto, Katsuhiko

    2008-01-01

    Establishing the occurrence of endocytosis in filamentous fungi was elusive in the past mainly due to the lack of reliable indicators of endocytosis. Recently, however, it was shown that the fluorescent dye N-(3-triethylammoniumpropyl)-4-(p-diethyl-aminophenyl-hexatrienyl)pyridinium dibromide (FM4-64) and the plasma membrane protein AoUapC (Aspergillus oryzae UapC) fused to enhanced green fluorescent protein (EGFP) were internalized from the plasma membrane by endocytosis. Although the occurr...

  17. A membrane fusion protein αSNAP is a novel regulator of epithelial apical junctions.

    Directory of Open Access Journals (Sweden)

    Nayden G Naydenov

    Full Text Available Tight junctions (TJs and adherens junctions (AJs are key determinants of the structure and permeability of epithelial barriers. Although exocytic delivery to the cell surface is crucial for junctional assembly, little is known about the mechanisms controlling TJ and AJ exocytosis. This study was aimed at investigating whether a key mediator of exocytosis, soluble N-ethylmaleimide sensitive factor (NSF attachment protein alpha (αSNAP, regulates epithelial junctions. αSNAP was enriched at apical junctions in SK-CO15 and T84 colonic epithelial cells and in normal human intestinal mucosa. siRNA-mediated knockdown of αSNAP inhibited AJ/TJ assembly and establishment of the paracellular barrier in SK-CO15 cells, which was accompanied by a significant down-regulation of p120-catenin and E-cadherin expression. A selective depletion of p120 catenin effectively disrupted AJ and TJ structure and compromised the epithelial barrier. However, overexpression of p120 catenin did not rescue the defects of junctional structure and permeability caused by αSNAP knockdown thereby suggesting the involvement of additional mechanisms. Such mechanisms did not depend on NSF functions or induction of cell death, but were associated with disruption of the Golgi complex and down-regulation of a Golgi-associated guanidine nucleotide exchange factor, GBF1. These findings suggest novel roles for αSNAP in promoting the formation of epithelial AJs and TJs by controlling Golgi-dependent expression and trafficking of junctional proteins.

  18. Calcium is not involved in the cAMP-mediated stimulation of Cl- conductance in the apical membrane of Necturus gallbladder epithelium.

    Science.gov (United States)

    Kottra, G

    1995-03-01

    The permeability properties of the forskolin-stimulated Cl- conductance in the apical membrane of Necturus gallbladder epithelium and the possible participation of intracellular Ca2+ in its stimulation have been investigated. The anion selectivity sequence as derived from biionic potential measurements (SCN- > I- approximately NO3- > Br- > Cl- > ISE-) differed from the sequence derived from measurements of apical membrane resistance (NO3- approximately Br- approximately Cl- > SCN- > I- approximately ISE-). Accordingly, the conductance was inhibited by SCN- and I- which, from the potential measurements, appeared to be more permeable than Cl-. This finding agrees with observations of the cystic fibrosis transmembrane conductance regulator (CFTR) Cl- channel reported recently. However, none of the commonly used Cl- channel blockers, such as 5-nitro-2-(3-phenylpropylamino)-benzoic acid (NPPB), 4,4'-diisothiocyanatostilbene-2,2'-disulphonic acid (DIDS), anthracene-9-carboxylic acid (9-AC) and glibenclamide reduced this conductance in Necturus gallbladder. In contrast to the situation in most other epithelia, elevation of intracellular Ca2+ concentration ([Ca2+]i) by ionomycin stimulated only K+ conductance and not that of Cl- in the apical cell membrane. Chelation of intracellular Ca2+ did not prevent the stimulation of Cl- conductance by forskolin. This indicates that [Ca2+]i does not have even a permissive role in the cyclic adenosine monophosphate-(cAMP)-mediated stimulation process, as would have been expected if exocytosis was involved. Further evidence against the involvement of exocytosis in the stimulation process came from the observation that the stimulation was not associated with an increase in apical membrane capacitance and was not suppressed by disruption of the cytoskeleton by preincubation of the tissue with cytochalasin D. The data indicate that Necturus gallbladder epithelium contains homologues of the CFTR Cl- channel which reside permanently in the

  19. Analysis of sperm membrane antigens relevant to antisperm antibody using Western blot

    Institute of Scientific and Technical Information of China (English)

    WangHF

    2002-01-01

    Objective:To identify the sperm membrane antigens associated with antisperm antibody.Methods:The antisperm antibody in serum was tested by ELISA.Antisperm antibody positive sera from 18 infertile men and 15 infertile women were used.The molecular weight(MW) of sperm membrane antigens associated with the antisperm antibody was analyzed with antisperm antibody positive serum using Western blot.Results:Eight kinds of MW of sperm membrane antigens were identified.The ratio of identification on the 78 KD(60.7%),60KD(71.4%),51KD(14.9%) and 23KD(14.29%)sperm antigen was higher than other.Conclusion:sperm membrane antigens with MW of 78KD,60KD,51KD and 23KD were associated with antisperm antibody and immunological infertility.

  20. Apical Scaffolding Protein NHERF2 Modulates the Localization of Alternatively Spliced Plasma Membrane Ca2+ Pump 2B Variants in Polarized Epithelial Cells*

    OpenAIRE

    Padányi, Rita; Xiong, Yuning; Antalffy, Géza; Lór, Krisztina; Pászty, Katalin; STREHLER, EMANUEL E.; Enyedi, Ágnes

    2010-01-01

    The membrane localization of the plasma membrane Ca2+-ATPase isoform 2 (PMCA2) in polarized cells is determined by alternative splicing; the PMCA2w/b splice variant shows apical localization, whereas the PMCA2z/b and PMCA2x/b variants are mostly basolateral. We previously reported that PMCA2b interacts with the PDZ protein Na+/H+ exchanger regulatory factor 2 (NHERF2), but the role of this interaction for the specific membrane localization of PMCA2 is not known. Here we show that co-expressio...

  1. Simple solid-phase radioimmunoassay for human leukemia-associated cell membrane antigens

    International Nuclear Information System (INIS)

    In the present study, a simple solid-phase radioimmunoassay was developed to determine detergent-extracted human leukemia-associated cell membrane antigens. In the assay, 96-well microtiter plates are coated with human leukemia cell membrane antigens containing a T cell leukemia or a non-T cell leukemia antigen in the presence of a detergent, and treated with 1.6% bovine serum albumin solution. The coated antigens were reacted with an appropriate murine monoclonal antibody (mAb). The bound mAb is determined by a second reaction with 125I-labeled F(ab')2 of goat anti-mouse Ig. The best antigen dose-dependent antibody binding results were obtained using the plates coated with antigens in the presence of taurocholate. In addition, the usefulness of the present assay with taurocholate during the purification of the antigens was demonstrated. (Auth.)

  2. Antigen presentation by liposomes bearing class II MHC and membrane IL-1.

    OpenAIRE

    Bakouche, O; LACHMAN, L.B.

    1990-01-01

    Liposomes containing membrane IL-1, Iak, and the antigen conalbumin were evaluated as "synthetic antigen presenting cells." The role of these three molecules in macrophage-T cell interaction was studied by testing their ability to induce the proliferation of a T-cell clone specific to conalbumin (the D10 cell line) or immune spleen cells sensitized three times in vivo with conalbumin. In the latter case, splenic macrophages were eliminated by adherence and a lysomotropic agent. The antigen co...

  3. NHERF1/EBP50 Controls Morphogenesis of 3D Colonic Glands by Stabilizing PTEN and Ezrin-Radixin-Moesin Proteins at the Apical Membrane

    Directory of Open Access Journals (Sweden)

    Maria-Magdalena Georgescu

    2014-04-01

    Full Text Available Na+/H+ exchanger 3 regulating factor 1/ezrin-radixin-moesin (ERM–binding phosphoprotein 50 (NHERF1/EBP50, an adaptor molecule that interacts with the ERM–neurofibromatosis type 2 family of cytoskeletal proteins through its ERM-binding region and with phosphatase and tensin homolog (PTEN and β-catenin through its PDZ domains, has been recently implicated in the progression of various human malignancies, including colorectal cancer (CRC. We report here that NHERF1 controls gland morphogenesis, as demonstrated in three-dimensional (3D human intestinal glands developing from a single nonpolarized cell. Starting from the early two-cell developmental stage, NHERF1 concentrates at the cellular interface in a central membrane disc that marks the apical pole delimiting the forming lumen. NHERF1 depletion leads to severe disruption of the apical-basal polarity, with formation of enlarged and distorted cell spheroids devoid of a central lumen. This characteristic and the increased number of mitoses in NHERF1-depleted spheroids, including multipolar ones, mimic high-grade dysplasia lesions observed in CRC progression. NHERF1 ERM-binding or PDZ-domain mutants fail to localize apically and impair gland formation most likely by outcompeting endogenous ligands, with the latter mutant completely aborting gland development. Examination of NHERF1 ligands showed that even if both ezrin and moesin colocalized with NHERF1 at the apical membrane, moesin but not ezrin depletion disrupted morphogenesis similarly to NHERF1. NHERF1 depletion resulted also in membrane displacement of PTEN and nuclear translocation of β-catenin, events contributing to polarity loss and increased proliferation. These findings reveal an essential role of NHERF1 in epithelial morphogenesis and polarity and validate this 3D system for modeling the molecular changes observed in CRC.

  4. Epithelial membrane antigen in cells from the uterine cervix: immunocytochemical staining of cervical smears.

    OpenAIRE

    Valkova, B; Ormerod, M G; Moncrieff, D.; Coleman, D V

    1984-01-01

    Smears made from cervical scrapes have been stained immunocytochemically for epithelial membrane antigen using a polyclonal antiserum and two monoclonal antibodies. With the polyclonal antiserum malignant cells and those showing dysplasia consistently expressed the antigen. Normal cells were generally negative, with the exception of some metaplastic cells. The monoclonal antibodies, although they stained the abnormal cells less consistently, gave the same pattern of staining. All three antibo...

  5. Antigenicity and immunogenicity of an extract from the cell wall and cell membrane of Histoplasma capsulatum yeast cells.

    OpenAIRE

    Gómez, A M; Rhodes, J C; Deepe, G S

    1991-01-01

    In order to identify T-cell antigens from Histoplasma capsulatum yeast cells, we prepared a detergent extract of the cell wall and cell membrane of yeast-phase H. capsulatum G217B and analyzed its antigenicity and immunogenicity. Mice injected with viable H. capsulatum yeast cells or with 500 or 1,000 micrograms of the extract mounted a delayed-type hypersensitivity response to solubilized cell wall and cell membrane. Vaccination with this antigenic preparation conferred a protective immune r...

  6. M-Type Phospholipase A2 Receptor as Target Antigen in Idiopathic Membranous Nephropathy

    Science.gov (United States)

    Beck, Laurence H.; Bonegio, Ramon G.B.; Lambeau, Gérard; Beck, David M.; Powell, David W.; Cummins, Timothy D.; Klein, Jon B.; Salant, David J.

    2009-01-01

    BACKGROUND Idiopathic membranous nephropathy, a common form of the nephrotic syndrome, is an antibody-mediated autoimmune glomerular disease. Serologic diagnosis has been elusive because the target antigen is unknown. METHODS We performed Western blotting of protein extracts from normal human glomeruli with serum samples from patients with idiopathic or secondary membranous nephropathy or other proteinuric or autoimmune diseases and from normal controls. We used mass spectrometry to analyze the reactive protein bands and confirmed the identity and location of the target antigen with a monospecific antibody. RESULTS Serum samples from 26 of 37 patients (70%) with idiopathic but not secondary membranous nephropathy specifically identified a 185-kD glycoprotein in non-reduced glomerular extract. Mass spectrometry of the reactive protein band detected the M-type phospholipase A2 receptor (PLA2R). Reactive serum specimens recognized recombinant PLA2R and bound the same 185-kD glomerular protein as did the monospecific anti-PLA2R antibody. Anti-PLA2R autoantibodies in serum samples from patients with membranous nephropathy were mainly IgG4, the predominant immunoglobulin subclass in glomerular deposits. PLA2R was expressed in podocytes in normal human glomeruli and colocalized with IgG4 in immune deposits in glomeruli of patients with membranous nephropathy. IgG eluted from such deposits in patients with idiopathic membranous nephropathy, but not in those with lupus membranous or IgA nephropathy, recognized PLA2R. CONCLUSIONS A majority of patients with idiopathic membranous nephropathy have antibodies against a conformation-dependent epitope in PLA2R. PLA2R is present in normal podocytes and in immune deposits in patients with idiopathic membranous nephropathy, indicating that PLA2R is a major antigen in this disease. PMID:19571279

  7. Library of monoclonal antibodies against brush border membrane epithelial antigens

    Energy Technology Data Exchange (ETDEWEB)

    Behar, M.; Katz, A.; Silverman, M.

    1986-03-01

    A purified fraction of proximal tubule brush border membranes (BBM) was prepared from dog kidney and used to immunize mice. The standard technique of hybridoma production was followed as described by Kohler and Milstein. Production of antibodies was detected by indirect immunofluorescence on dog kidney slices and by immunodot against the purified fraction on nitrocellulose. Five hybrids exhibited anti BBM activity. These were cloned twice and yielded stable cell lines producing IgG type monoclonal antibodies against BBM. They were designated A/sub 1/, C/sub 7/, D/sub 3/, D/sub 7/ and H/sub 4/. As a family these five monoclonals have broad tissue specificity, i.e. positive staining of the surface mucosa of intestinal kidney proximal tubules. D/sub 3/ exhibits even broader specificity for epithelium reacting with bile canaliculi and choroid plexus. The authors have verified that at least 4/5 antibodies are directed against BBM protein as revealed by immunoprecipitation of solubilized BBM and detected by Coomassie blue staining or autoradiography of lactoperoxidase labelled BBM. Most interestingly all antibodies bind to the surface of LL CPK/sub 1/ cells, a continuous pig kidney cell line of undefined origin but exhibiting many characteristics of proximal tubule cells. The library of monoclonal antibodies obtained provide important probes with which to study membrane biogenesis and polarization in epithelial cells.

  8. CM2 antigen, a potential novel molecule participating in glucuronide transport on rat hepatocyte canalicular membrane

    Directory of Open Access Journals (Sweden)

    L. Wang

    2012-06-01

    Full Text Available The polarized molecules predominately distributing at hepatocyte canalicular surface play a vital role in disclosing the process of bile formation and etiopathogenisis of cholestatic live diseases. Therefore, it is important to find novel polarized molecules on hepatocyte canalicular membrane. In the present study, canalicular membrane vesicles (CMVs isolated from rat hepatocyte by density gradient centrifugation were used as immunogens to produce hybridoma and 46 strains of monoclonal antibodies (mAb against CMVs were obtained. With a series of morphological assay methods, including immunohistochemistry, immunofluorescence and immuno-electron microscope, the antigens recognized by canalicular mAb1 (CM1 and canalicular mAb2 (CM2 were confirmed to predominately distribute at hepatocyte canalicular membrane. Transport activity assay revealed that CM2 could inhibit ATP-dependent E217βG uptake of rat hepatocyte CMVs. Meanwhile, Western blotting analysis showed that the molecular mass of CM2 antigen was approximately 110kDa, which was much less than Mr 180kDa of multidrug resistance-associated protein 2 (MRP2 involved in glucuronide transport. These data indicated that CM2 antigen might be a potential novel molecule participating in glucuronide transport on the hepatocyte canalicular membrane.

  9. The effects of acellular amniotic membrane matrix on osteogenic differentiation and ERK1/2 signaling in human dental apical papilla cells.

    Science.gov (United States)

    Chen, Yi-Jane; Chung, Min-Chun; Jane Yao, Chung-Chen; Huang, Chien-Hsun; Chang, Hao-Hueng; Jeng, Jiiang-Huei; Young, Tai-Horng

    2012-01-01

    The amniotic membrane (AM) has been widely used in the field of tissue engineering because of the favorable biological properties for scaffolding material. However, little is known about the effects of an acellular AM matrix on the osteogenic differentiation of mesenchymal stem cells. In this study, it was found that both basement membrane side and collagenous stroma side of the acellular AM matrix were capable of providing a preferential environment for driving the osteogenic differentiation of human dental apical papilla cells (APCs) with proven stem cell characteristics. Acellular AM matrix potentiated the induction effect of osteogenic supplements (OS) such as ascorbic acid, β-glycerophosphate, and dexamethasone and enhanced the osteogenic differentiation of APCs, as seen by increased core-binding factor alpha 1 (Cbfa-1) phosphorylation, alkaline phosphatase activity, mRNA expression of osteogenic marker genes, and mineralized matrix deposition. Even in the absence of soluble OS, acellular AM matrix also could exert the substrate-induced effect on initiating APCs' differentiation. Especially, the collagenous stroma side was more effective than the basement membrane side. Moreover, the AM-induced effect was significantly inhibited by U0126, an inhibitor of extracellular signaling-regulated kinase 1/2 (ERK1/2) signaling. Taken together, the osteogenic differentiation promoting effect on APCs is AM-specific, which provides potential applications of acellular AM matrix in bone/tooth tissue engineering.

  10. Inhibitory effect of calcium on non-heme iron absorption may be related to translocation of DMT-1 at the apical membrane of enterocytes.

    Science.gov (United States)

    Thompson, Ben A V; Sharp, Paul A; Elliott, Ruan; Fairweather-Tait, Susan J

    2010-07-28

    Many studies show that calcium reduces iron absorption from single meals, but the underlying mechanism is not known. We tested the hypothesis that calcium alters the expression and/or functionality of iron transport proteins. Differentiated Caco-2 cells were treated with ferric ammonium citrate and calcium chloride, and ferritin, DMT-1, and ferroportin were quantified in whole-cell lysate and cell-membrane fractions. Calcium attenuated the iron-induced increase in cell ferritin levels in a dose-dependent manner; a significant decrease was seen at calcium concentrations of 1.25 and 2.5 mM but was only evident after a 16-24 h incubation period. Calcium and iron treatments decreased DMT-1 protein in Caco-2 cell membranes, although total DMT-1 in whole cell lysates was unchanged by either iron or calcium. No change was seen in ferroportin expression. Our data suggest that calcium reduces iron bioavailability by decreasing DMT-1 expression at the apical cell membrane, thereby downregulating iron transport into the cell.

  11. Common channels for water and protons at apical and basolateral cell membranes of frog skin and urinary bladder epithelia. Effects of oxytocin, heavy metals, and inhibitors of H(+)-adenosine triphosphatase

    Energy Technology Data Exchange (ETDEWEB)

    Harvey, B.; Lacoste, I.; Ehrenfeld, J. (Commissariat a l' Energie Atomique, Villefranche-sur-mer (France))

    1991-04-01

    We have compared the response of proton and water transport to oxytocin treatment in isolated frog skin and urinary bladder epithelia to provide further insights into the nature of water flow and H+ flux across individual apical and basolateral cell membranes. In isolated spontaneous sodium-transporting frog skin epithelia, lowering the pH of the apical solution from 7.4 to 6.4, 5.5, or 4.5 produced a fall in pHi in principal cells which was completely blocked by amiloride, indicating that apical Na+ channels are permeable to protons. When sodium transport was blocked by amiloride, the H+ permeability of the apical membranes of principal cells was negligible but increased dramatically after treatment with antidiuretic hormone (ADH). In the latter condition, lowering the pH of the apical solution caused a voltage-dependent intracellular acidification, accompanied by membrane depolarization, and an increase in membrane conductance and transepithelial current. These effects were inhibited by adding Hg2+ (100 microM) or dicyclohexylcarbodiimide (DCCD, 10(-5) M) to the apical bath. Net titratable H+ flux across frog skin was increased from 30 +/- 8 to 115 +/- 18 neq.h-1.cm-2 (n = 8) after oxytocin treatment (at apical pH 5.5 and serosal pH 7.4) and was completely inhibited by DCCD (10(-5) M). The basolateral membranes of the principal cells in frog skin epithelium were found to be spontaneously permeable to H+ and passive electrogenic H+ transport across this membrane was not affected by oxytocin. Lowering the pH of the basolateral bathing solution (pHb) produced an intracellular acidification and membrane depolarization (and an increase in conductance when the normal dominant K+ conductance of this membrane was abolished by Ba2+ 1 mM). These effects of low pHb were blocked by micromolar concentrations of heavy metals (Zn2+, Ni2+, Co2+, Cd2+, and Hg2+).

  12. Development and heterogeneity of antigens in the immature nephron. Reactivity with human antiglomerular basement membrane autoantibodies.

    OpenAIRE

    Jeraj, K.; Fish, A. J.; Yoshioka, K; Michael, A. F.

    1984-01-01

    Indirect immunofluorescence microscopy was performed with 15 human anti-glomerular basement membrane (GBM) antibodies and mouse monoclonal antibodies to Type IV collagen (MBM4) and renal basement membranes (MBM15) on renal tissue from 6 fetuses (gestational age, 15-23 weeks), 8 infants (age, 1-21 days), and 8 children and adults (ages, 3-27 years). Of the 15 human anti-GBM antibodies that react with GBM in adult glomeruli, only 4 identified antigens in the GBM of fetal and infant glomeruli. I...

  13. Intravacuolar Membranes Regulate CD8 T Cell Recognition of Membrane-Bound Toxoplasma gondii Protective Antigen

    OpenAIRE

    Jodie Lopez; Amina Bittame; Céline Massera; Virginie Vasseur; Grégory Effantin; Anne Valat; Célia Buaillon; Sophie Allart; Barbara A. Fox; Leah M. Rommereim; David J. Bzik; Guy Schoehn; Winfried Weissenhorn; Jean-François Dubremetz; Jean Gagnon

    2015-01-01

    Apicomplexa parasites such as Toxoplasma gondii target effectors to and across the boundary of their parasitophorous vacuole (PV), resulting in host cell subversion and potential presentation by MHC class I molecules for CD8 T cell recognition. The host-parasite interface comprises the PV limiting membrane and a highly curved, membranous intravacuolar network (IVN) of uncertain function. Here, using a cell-free minimal system, we dissect how membrane tubules are shaped by the parasite effecto...

  14. Binding of Streptococcus mutans antigens to heart and kidney basement membranes.

    Science.gov (United States)

    Stinson, M W; Barua, P K; Bergey, E J; Nisengard, R J; Neiders, M E; Albini, B

    1984-01-01

    Using indirect immunofluorescence, alkali-extracted components of Streptococcus mutans were found to bind in vitro to capillary walls and sarcolemmal sheaths of monkey cardiac muscle and to glomerular and tubular basement membranes of monkey kidney. Adsorption of S. mutans components to tissue fragments was also detected by indirect radioimmunoassay and immunoblotting on nitrocellulose paper. Antibodies did not bind to untreated, control tissues in these experiments, proving that antigens shared by S. mutans and tissue components were not involved. Rabbit and monkey heart and kidney components bound S. mutans antigens of 24,000, 35,000, and 65,000 Mr. Monkey heart also bound molecules of 90,000 and 120,000 Mr. Rabbits immunized by intravenous injection of disrupted S. mutans cells developed severe nephritis that was characterized by the deposition of immunoglobulins, complement component C3, and S. mutans antigens in the glomeruli. Immunoglobulin G eluted from nephritic kidneys reacted in immunoblots with the 24,000, 35,000, and 65,000 Mr components of S. mutans extract, indicating that the antigens that bound to tissue in vitro also bound in vivo and reacted with antibodies in situ. Antibodies to other S. mutans antigens were not detected in the kidney eluate, although they were present in the serum of the same rabbit. Images PMID:6384042

  15. Protein-mediated inward translocation of phospholipids occurs in both the apical and basolateral plasma membrane domains of epithelial cells

    NARCIS (Netherlands)

    Pomorski, T.; Herrmann, A.; Müller, P.; van Meer, G.F.B.P.; Burger, K.N.J.

    1999-01-01

    The translocation of spin-labeled analogues of phosphatidylcholine (4- doxylpentanoyl-PC, SLPC), phosphatidylethanolamine (SL-PE), phosphatidylserine (SL-PS), and sphingomyelin (SL-SM) from the outer to the inner leaflet of the plasma membrane bilayer was investigated in dog kidney MDCK II and human

  16. Effect of anticancer therapy on Tn antigen exposure on the leucocyte membranes in patients with leukemia

    Directory of Open Access Journals (Sweden)

    G. S. Maslak

    2014-08-01

    them with propidium iodide. The result was analyzed with FC Express. According to our data, Tn-antigen exposure was not detected on the surface of blood cells (lymphocytes, monocytes and granulocytes in the control group and in patients with polycythemia vera and subleukemic myelosis. Nevertheless, Tn-antigen was identified on the surface of more than 80% of lymphocytes in chronic lymphocytic leukemia patients. The intensity of this tumor-associated antigen exposure on lymphocytes membrane was 100 times higher compared with that in normal lymphocytes. In chronic lymphocytic leukemia patients after COP-treatment the number of lymphocytes with surface Tn-antigen was equal to 28,1 ± 0,8%, and after FC-treatment it decreased to 9,5 ± 0,5%. Moreover, positive effect of cytotoxic therapy used in treatment of patients with chronic lymphocytic leukemia on intensity of Tn-antigen exposure on the surface of lymphocytes was shown. FC-therapy (fludarabine, cyclophosphamide is more effective; compared with the data prior to this treatment it 40 times reduced the relevant index. Therefore, it can be applied in Ukraine for chemotherapeutic treatment schemes effective against Tn-antigen.

  17. Cross-reactivity of anti-H pylori antibodies with membrane antigens of human erythrocytes

    Institute of Scientific and Technical Information of China (English)

    Feng-Hua Guo; Fan-Ling Meng; Jian-Zhong Zhang; Xiao-Mei Yan; Chun-Xiang Fan; Fei Zhao; Yuan Hu; Di Xiao; Xun Zeng; Mao-Jun Zhang; Li-Hua He

    2007-01-01

    AIM: To investigate whether anti-H pylori antibodies have cross-reaction with antigens of erythrocyte membrane.METHODS: Blood samples were collected from 14 volunteers (8 positive and 6 negative for H pylori detected by 13C-urea breath test) of the general population. Erythrocyte membrane proteins of the subjects were examined by Western blot using antiH pylori serum. The proteins related to the positive bands were identified by mass spectrum analysis.RESULTS: Anti-H pylori antibodies had cross-reaction with the proteins of about 50 kDa of erythrocyte membranes in all samples independent of H pylori infection. One protein in the positive band was identified as Chain S, the crystal structure of the cytoplasmic domain of human erythrocyte Band-3 protein.CONCLUSION: Anti-H pylori antibodies cross-react with some antigens of human erythrocyte membrane, which may provide a clue for the relationship between H pylori infection and vascular disorders.

  18. Endodontic management of nonvital permanent teeth having immature roots with one step apexification, using mineral trioxide aggregate apical plug and autogenous platelet-rich fibrin membrane as an internal matrix: Case series.

    Science.gov (United States)

    Sharma, Vivek; Sharma, Sarang; Dudeja, Pooja; Grover, Shibani

    2016-01-01

    A tooth with blunderbuss canal and open apex can be an endodontic challenge because of difficulty in obtaining an apical seal, and existing thin radicular walls which are susceptible to fracture. To overcome the limitations of traditional long-term calcium hydroxide apexification procedures, nonsurgical one step apexification using an array of materials such as mineral trioxide aggregate (MTA) has been suggested. However, adequate compaction of MTA in teeth with wide open apices can be an arduous task, and an internal matrix is required for controlled placement of MTA against which obturating material can be condensed. Platelet-rich fibrin (PRF), a second generation platelet concentrate containing several growth factors that promotes hard and soft-tissue healing, has been used as an internal matrix to create an apical plug of MTA and hence prevent extrusion of filling materials. This case series presents the endodontic management of immature permanent teeth with open apices using internal matrix of autologous PRF membrane and one step apical barrier placement of MTA. PMID:27041904

  19. T-cell-specific membrane antigens in the Mexican axolotl (urodele amphibian).

    Science.gov (United States)

    Kerfourn, F; Guillet, F; Charlemagne, J; Tournefier, A

    1992-01-01

    Comparative analysis of SDS-PAGE patterns of axolotl spleen cells membrane detergent lysates showed important discrepancies between control and thymectomized animals. Among these, a 38-kD protein band, which appeared as a major protein in controls, was not or poorly expressed after thymectomy. A rabbit antiserum (L12) raised against the 38-kD eluted band labeled in indirect immunofluorescence 80-86% of thymocytes and 40-46% of mIg- lymphoid cells in the spleen. The anti-38-kD antibodies stained in Western blotting two antigenically related polypeptides of 38- and 36-kD on splenocyte membrane lysates. Two-dimensional NEPHGE-PAGE analysis indicated that the anti-38-kD antibodies reacted in the spleen with several gathered spots in the 7.8-8.2 pI range, corresponding to 38-36-kD microheterogeneous polypeptides. Most of these spots are not further expressed in thymectomized animals. These results support evidence that the 38-kD surface antigens can be considered as specific surface markers of the axolotl thymus-derived lymphocytes. PMID:1627952

  20. Production of a Shigella sonnei Vaccine Based on Generalized Modules for Membrane Antigens (GMMA, 1790GAHB.

    Directory of Open Access Journals (Sweden)

    Christiane Gerke

    Full Text Available Recently, we developed a high yield production process for outer membrane particles from genetically modified bacteria, called Generalized Modules of Membrane Antigens (GMMA, and the corresponding simple two step filtration purification, enabling economic manufacture of these particles for use as vaccines. Using a Shigella sonnei strain that was genetically modified to produce penta-acylated lipopolysaccharide (LPS with reduced endotoxicity and to maintain the virulence plasmid encoding for the immunodominant O antigen component of the LPS, scale up of the process to GMP pilot scale was straightforward and gave high yields of GMMA with required purity and consistent results. GMMA were formulated with Alhydrogel and were highly immunogenic in mice and rabbits. In mice, a single immunization containing 29 ng protein and 1.75 ng of O antigen elicited substantial anti-LPS antibody levels. As GMMA contain LPS and lipoproteins, assessing potential reactogenicity was a key aspect of vaccine development. In an in vitro monocyte activation test, GMMA from the production strain showed a 600-fold lower stimulatory activity than GMMA with unmodified LPS. Two in vivo tests confirmed the low potential for reactogenicity. We established a modified rabbit pyrogenicity test based on the European Pharmacopoeia pyrogens method but using intramuscular administration of the full human dose (100 μg of protein. The vaccine elicited an average temperature rise of 0.5°C within four hours after administration, which was considered acceptable and showed that the test is able to detect a pyrogenic response. Furthermore, a repeat dose toxicology study in rabbits using intramuscular (100 μg/dose, intranasal (80 μg/dose, and intradermal (10 μg/dose administration routes showed good tolerability of the vaccine by all routes and supported its suitability for use in humans. The S. sonnei GMMA vaccine is now in Phase 1 dose-escalation clinical trials.

  1. Protective efficacy of bacterial membranes containing surface-exposed BM95 antigenic peptides for the control of cattle tick infestations.

    Science.gov (United States)

    Canales, Mario; Labruna, Marcelo B; Soares, João F; Prudencio, Carlos R; de la Fuente, José

    2009-12-01

    The Rhipicephalus (Boophilus) microplus BM86 and BM95 glycoproteins are homologous proteins that protect cattle against tick infestations. In this study, we demonstrated that the recombinant chimeric protein comprising tick BM95 immunogenic peptides fused to the A. marginale MSP1a N-terminal region for presentation on the Escherichia coli membrane was protective against R. microplus infestations in rabbits. This system provides a novel and simple approach for the production of tick protective antigens by surface display of antigenic protein chimera on live E. coli and suggests the possibility of using recombinant bacterial membrane fractions for vaccination against cattle tick infestations. PMID:19835826

  2. Antigens of the basement membranes of the seminiferous tubules induce autoimmunity in Wistar rats.

    Science.gov (United States)

    Lustig, L; Satz, M L; Sztein, M B; Denduchis, B

    1982-05-01

    A preparation enriched in basement membranes from seminiferous tubules was isolated from rat testes (STBM) and injected with complete Freund's adjuvant into Wistar rats. In 60% of animals a mild multifocal orchitis was observed. In damaged areas, perivascular and peritubular mononuclear cell infiltrates and different degrees of cell sloughing of some seminiferous tubules were observed. Electron microscopy revealed focal thickenings and delamination of the basement membrane of the seminiferous tubules as well as vacuolization of Sertoli cell cytoplasm. Using immunofluorescence discontinuous linear deposits of IgG were detected along the seminiferous tubular wall. Moreover, the same pattern of immunofluorescence was observed when the IgG eluted from the testes of the immunized rats was layered on sections of normal rat testis. Circulating antibodies to STBM were detected using passive haemagglutination in approximately 45% of the immunized rats, with titers ranging from 1:20 to 1:80. Leukocyte migration was inhibited when the spleen cells of the immunized rats were incubated with antigens from the basement membrane of seminiferous tubules, whilst a negative reaction was obtained when the soluble fraction of testis homogenate was used. PMID:7050376

  3. Mixed enzyme-linked immunosorbent assay (MELISA) for HLA class I antigen: a plasma membrane marker.

    Science.gov (United States)

    Bjerrum, O W; Borregaard, N

    1990-03-01

    This study introduces a simple, reproducible assay for HLA class I antigen using antibodies against beta 2-microglobulin and the heavy chain on HLA. The sandwich technique was named mixed enzyme-linked immunosorbent assay (MELISA), and was designed for identification of plasma membranes in neutrophil subcellular fractions. The subcellular localization of HLA was identical to that of other plasma membrane markers, [3H]concanavalin A and detergent-independent alkaline phosphatase, and was unchanged by stimulation of cells by weak and strong secretagogues. In addition to the presence as part of the HLA complex in the plasma membrane uncomplexed beta 2-microglobulin is present in the specific granules of neutrophils. However, the release of beta 2-microglobulin from intact neutrophils stimulated with formyl-methionylleucylphenylalanine was much higher than could be explained by exocytosis of specific granules. Subcellular fractionation studies demonstrated that beta 2-microglobulin is localized in fractions characterized by latent alkaline phosphatase and released from this novel secretory compartment in response to stimulation with formyl-methionylleucylphenylalanine. PMID:2181625

  4. Control of tick infestations in cattle vaccinated with bacterial membranes containing surface-exposed tick protective antigens.

    Science.gov (United States)

    Almazán, Consuelo; Moreno-Cantú, Orlando; Moreno-Cid, Juan A; Galindo, Ruth C; Canales, Mario; Villar, Margarita; de la Fuente, José

    2012-01-01

    Vaccines containing the Rhipicephalus (Boophilus) microplus BM86 and BM95 antigens protect cattle against tick infestations. Tick subolesin (SUB), elongation factor 1a (EF1a) and ubiquitin (UBQ) are new candidate protective antigens for the control of cattle tick infestations. Previous studies showed that R. microplus BM95 immunogenic peptides fused to the Anaplasma marginale major surface protein (MSP) 1a N-terminal region (BM95-MSP1a) for presentation on the Escherichia coli membrane were protective against R. microplus infestations in rabbits. In this study, we extended these results by expressing SUB-MSP1a, EF1a-MSP1a and UBQ-MSP1a fusion proteins on the E. coli membrane using this system and demonstrating that bacterial membranes containing the chimeric proteins BM95-MSP1a and SUB-MSP1a were protective (>60% vaccine efficacy) against experimental R. microplus and Rhipicephalus annulatus infestations in cattle. This system provides a novel, simple and cost-effective approach for the production of tick protective antigens by surface display of antigenic protein chimera on the E. coli membrane and demonstrates the possibility of using recombinant bacterial membrane fractions in vaccine preparations to protect cattle against tick infestations. PMID:22085549

  5. Identification of the cutaneous basement membrane zone antigen and isolation of antibody in linear immunoglobulin A bullous dermatosis.

    OpenAIRE

    Zone, J J; Taylor, T B; Kadunce, D P; Meyer, L J

    1990-01-01

    Linear IgA bullous dermatosis (LABD) is a rare blistering skin disease characterized by basement membrane zone deposition of IgA. This study identifies a tissue antigen detected by patient serum and then isolates the autoantibody using epidermis and protein bands blotted on nitrocellulose as immunoabsorbents. Sera from 10 patients (9 with cutaneous disease and 1 with cicatrizing conjunctivitis) were evaluated. Indirect immunofluorescence revealed an IgA anti-basement membrane antibody in 6 of...

  6. Prostate specific membrane antigen- a target for imaging and therapy with radionuclides

    DEFF Research Database (Denmark)

    Bouchelouche, Kirsten; Choyke, Peter L; Capala, Jacek

    2010-01-01

    Prostate cancer continues to represent a major health problem, and yet there is no effective treatment available for advanced metastatic disease. Thus, there is an urgent need for the development of more effective treatment modalities that could improve the outcome. Because prostate specific...... membrane antigen (PSMA), a transmembrane protein, is expressed by virtually all prostate cancers, and its expression is further increased in poorly differentiated, metastatic, and hormone-refractory carcinomas, it is a very attractive target. Molecules targeting PSMA can be labelled with radionuclides...... to become both diagnostic and/or therapeutic agents. The use of PSMA binding agents, labelled with diagnostic and therapeutic radio-isotopes, opens up the potential for a new era of personalized management of metastatic prostate cancer....

  7. The Expression of Sperm Membrane Peptide-Hepatitis B Surface Antigen Fusion Protein with Recombinant Vaccinia Virus

    Institute of Scientific and Technical Information of China (English)

    杨晓鸣; 赵峰; 严缘昌; 李光地; 汪垣

    1998-01-01

    A synthetic oligonucleotide, HSD-2a, encoding a peptide segment of the extracellular domain of a human sperm membrane protein, YWK-Ⅱ, was fused with hepatitis B surface antigen gene (HBs gene). The fused gene was then cloned to pUC18 plasmid.

  8. Development of a Novel, Ultra-rapid Biosensor for the Qualitative Detection of Hepatitis B Virus-associated Antigens and Anti-HBV, Based on “Membrane-engineered” Fibroblast Cells with Virus-Specific Antibodies and Antigens

    OpenAIRE

    Antonios Perdikaris; Nikos Alexandropoulos; Spiridon Kintzios

    2009-01-01

    A novel miniature cell biosensor detection system for the detection of Hepatis B virus (HBV)-associated antigens and anti-HBV is described. The biosensor is based on “membrane-engineered” Vero fibroblast cells immobilized in an alginate matrix. The membrane-engineering process involved the electroinsertion of anti-HBV specific antibodies (anti-HBs, anti-HBe) or antigens (HBsAg) in the membranes of the Vero cells. The attachment of a homologous antigen to the electroinserted antibody (or, resp...

  9. 人鼻粘膜上皮细胞Na+通道的初步研究%Sodium channels in the apical membrane of human nasal epithelial cells

    Institute of Scientific and Technical Information of China (English)

    张欣欣; 郭永清; 董震; 杨占泉; 张文杰

    2001-01-01

    Objective To study the electrophysiological properties of sodium channels in the apical membrane of human nasal epithelial cells. Method Nasal epithelial cells of human inferior turbinate from patients with obstructive sleep apnea syndrome were cultured in serum free medium on collagen gel-coated membranes at an air-liquid interface and studied by a patch clamp technique. Results In cell-attached patches, a typical single channel current with a conductance of 21.09pS and reversal potential of -50.96 were recorded. The permeability ratio PNa/PK was more than 5.80. In the presence of 10-4 mmol/L amiloride in the pipette, the incidence of sodium channels decreased from 26.67% to 5.13%. This revealed that a population of channels were inhibited by amiloride at a dose of 10-4 mmol/L. Ca2+ at dose of 10-3 mmol/L did not influence the incidence of sodium channels. There was no obvious association between voltage and the open probability of the channels. Conclusions Our results indicate that most Na+ channels in cell-attached patches of human nasal epithelial cells are amiloride-sensitive and Na+ selective. Only a few channels are amiloride-insensitive. The channels were not activated by extracellular Ca2+ and the open probability followed a voltage-independent manner.%目的 明确人鼻粘膜上皮细胞Na+通道的特性,为研究Na+通道在鼻粘膜病理性改变及治疗中的作用奠定理论基础。 方法 利用膜片钳技术对经无血清气-液界面培养的鼻源性鼾症患者手术切除下鼻甲标本的鼻上皮细胞进行Na+通道基本特性研究。 结果 在细胞贴附式膜片上,可记录到典型的单通道电流,其电导为21.09pS,反转电位为-50.96mV,且77.78%反转电位5.80。在Na+通道抑制剂10-4 mmol/L Amiloride存在于电极液内时,Na+通道发生率从26.7%减少到5.13%(P0.05)。电压对开放概率无明显影响。 结论 在细胞贴附式膜片上,人鼻粘膜上皮细胞具有大

  10. Bacterial histo-blood group antigens contributing to genotype-dependent removal of human noroviruses with a microfiltration membrane.

    Science.gov (United States)

    Amarasiri, Mohan; Hashiba, Satoshi; Miura, Takayuki; Nakagomi, Toyoko; Nakagomi, Osamu; Ishii, Satoshi; Okabe, Satoshi; Sano, Daisuke

    2016-05-15

    We demonstrated the genotype-dependent removal of human norovirus particles with a microfiltration (MF) membrane in the presence of bacteria bearing histo-blood group antigens (HBGAs). Three genotypes (GII.3, GII.4, and GII.6) of norovirus-like particles (NoVLPs) were mixed with three bacterial strains (Enterobacter sp. SENG-6, Escherichia coli O86:K61:B7, and Staphylococcus epidermidis), respectively, and the mixture was filtered with an MF membrane having a nominal pore size of 0.45 μm. All NoVLP genotypes were rejected by the MF membrane in the presence of Enterobacter sp. SENG-6, which excreted HBGAs as extracellular polymeric substances (EPS). This MF membrane removal of NoVLPs was not significant when EPS was removed from cells of Enterobacter sp. SENG-6. GII.6 NoVLP was not rejected with the MF membrane in the presence of E. coli O86:K61:B7, but the removal of EPS of E. coli O86:K61:B7 increased the removal efficiency due to the interaction of NoVLPs with the exposed B-antigen in lipopolysaccharide (LPS) of E. coli O86:K61:B7. No MF membrane removal of all three genotypes was observed when S. epidermidis, an HBGA-negative strain, was mixed with NoVLPs. These results demonstrate that the location of HBGAs on bacterial cells is an important factor in determining the genotype-dependent removal efficiency of norovirus particles with the MF membrane. The presence of HBGAs in mixed liquor suspended solids from a membrane bioreactor (MBR) pilot plant was confirmed by immune-transmission electron microscopy, which implies that bacterial HBGAs can contribute to the genotype-dependent removal of human noroviruses with MBR using MF membrane. PMID:27095709

  11. Uptake of triclopyr (3,5,6-trichloro-2-pyridinyloxyacetic acid) and dicamba (3,6-dichloro-2-methoxybenzoic acid) from the apical membranes of the human intestinal Caco-2 cells.

    Science.gov (United States)

    Kimura, Osamu; Tsukagoshi, Kensuke; Hayasaka, Moriaki; Endo, Tetsuya

    2012-01-01

    We investigated whether the uptake of triclopyr (3, 5, 6-trichloro-2-pyridinyloxyacetic acid) and dicamba (3,6-dichloro-2-methoxybenzoic acid) across the apical membrane of Caco-2 cells was mediated via proton-linked monocarboxylic acid transporters (MCTs). The uptake of triclopyr from the apical membranes was fast, pH-, temperature-, and concentration dependent, required metabolic energy to proceed, and was competitively inhibited by monocarboxylic acids such as benzoic acid and ferulic acid (substrates of L-lactic acid-insensitive MCTs), but not by L-lactic acid. Thus, the uptake of triclopyr in Caco-2 cells appears to be mediated mainly via L-lactic acid-insensitive MCTs. In contrast, the uptake of dicamba (a benzoic acid derivative) was slow, and it was both pH- and temperature dependent. Coincubation with ferulic acid did not decrease the uptake of dicamba, although coincubation with benzoic acid moderately decreased it. The uptake of dicamba appears to be mediated mainly via passive diffusion, which is in contrast to the uptake of benzoic acid via MCTs. We speculate that the substituted groups in dicamba may inhibit uptake via MCTs. PMID:21766207

  12. Creatinine clearance, urinary excretion of glomerular basement membrane antigens and renal histology in congenital nephrotic syndrome of Finnish type.

    Science.gov (United States)

    Huttunen, N P

    1977-04-01

    The endogenous creatinine clearance and urinary excretion rate of glomerular basement membrane (GBM) antigens were followed from 2 to 19 months in fifteen patients with congenital nephrotic syndrome (CNF). The quantitative examination of renal morphology was made on fourteen of these patients. Creatinine clearance increased during the first few months of life and thereafter gradually decreased. The urinary excretion rate of GBM antigens rose during the course of the disease. The creatinine clearance did not correlate significantly with glomerular fibrosis but it did correlate with tubular atrophy and interstitial fibrosis. The urinary excretion of GBM antigens correlated significantly with glomerular and interstitial fibrosis and with tubular atrophy. It is concluded that there is a clear progress in the disease and the renal histological changes probably are caused by accumulation of GBM material in glomeruli.

  13. Nephritogenic antigen determinants in epidermal and renal basement membranes of kindreds with Alport-type familial nephritis.

    OpenAIRE

    Kashtan, C; Fish, A. J.; Kleppel, M; Yoshioka, K; Michael, A. F.

    1986-01-01

    We probed epidermal basement membranes (EBM) of acid-urea denatured skin from members of kindreds with Alport-type familial nephritis (FN) for the presence of antigens reactive with Goodpasture sera (GPS) and serum (FNS) from an Alport patient who developed anti-glomerular basement membrane (GBM) nephritis in a renal allograft. By immunoblotting, GPS reacted primarily with the 28,000 molecular weight (mol wt) monomer but also the 24,000 mol wt and 26,000 mol wt monomers of the noncollagenous ...

  14. Carcinoembryonic antigen is anchored to membranes by covalent attachment to a glycosylphosphatidylinositol moiety: identification of the ethanolamine linkage site.

    OpenAIRE

    Hefta, S A; Hefta, L J; Lee, T.D. (Taunia D.); Paxton, R J; Shively, J. E.

    1988-01-01

    The COOH-terminal amino acid of carcinoembryonic antigen (CEA) is shown to covalently link with ethanolamine, evidence consistent with the anchorage of CEA to the plasma membrane through a phosphatidylinositol-glycan tail. Purified CEA was digested with trypsin, and the resulting peptides were isolated by reverse-phase HPLC. Tryptic hexapeptide T12, terminating atypically with alanine, corresponded in sequence (Ser-Ile-Thr-Val-Ser-Ala) with the last six residues (637-642) of the third repeati...

  15. Development of a Novel, Ultra-rapid Biosensor for the Qualitative Detection of Hepatitis B Virus-associated Antigens and Anti-HBV, Based on “Membrane-engineered” Fibroblast Cells with Virus-Specific Antibodies and Antigens

    Directory of Open Access Journals (Sweden)

    Antonios Perdikaris

    2009-03-01

    Full Text Available A novel miniature cell biosensor detection system for the detection of Hepatis B virus (HBV-associated antigens and anti-HBV is described. The biosensor is based on “membrane-engineered” Vero fibroblast cells immobilized in an alginate matrix. The membrane-engineering process involved the electroinsertion of anti-HBV specific antibodies (anti-HBs, anti-HBe or antigens (HBsAg in the membranes of the Vero cells. The attachment of a homologous antigen to the electroinserted antibody (or, respectively, of the antibody to the electroinserted antigen triggered specific changes to the cell membrane potential that were measured by appropriate microelectrodes, according to the principle of the Bioelectric Recognition Assay (BERA. The sensor was used for screening 133 clinical blood serum samples according to a double-blind protocol. Considerably higher sensor responses were observed against HBV-positive samples, compared with responses against negative samples or samples positive for heterologous hepatitis viruses such as Hepatitis C (HCV virus. Detection of anti-HBs antibodies was made possible by using a biosensor based on immobilized Vero cells bearing the respective antigen (HBsAg. The observed response was rapid (45 sec and quite reproducible. Fluorescence microscopy observations showed that attachment of HBV particles to cells membrane-engineered with anti-HBs was associated with a decrease of [Ca2+]cyt. The perspectives for using the novel biosensor as a qualitative, rapid screening, high throughput assay for HBV antigens and anti-HBs in clinical samples is discussed.

  16. Prostate specific membrane antigen (PSMA regulates angiogenesis independently of VEGF during ocular neovascularization.

    Directory of Open Access Journals (Sweden)

    Christina L Grant

    Full Text Available BACKGROUND: Aberrant growth of blood vessels in the eye forms the basis of many incapacitating diseases and currently the majority of patients respond to anti-angiogenic therapies based on blocking the principal angiogenic growth factor, vascular endothelial growth factor (VEGF. While highly successful, new therapeutic targets are critical for the increasing number of individuals susceptible to retina-related pathologies in our increasingly aging population. Prostate specific membrane antigen (PSMA is a cell surface peptidase that is absent on normal tissue vasculature but is highly expressed on the neovasculature of most solid tumors, where we have previously shown to regulate angiogenic endothelial cell invasion. Because pathologic angiogenic responses are often triggered by distinct signals, we sought to determine if PSMA also contributes to the pathologic angiogenesis provoked by hypoxia of the retina, which underlies many debilitating retinopathies. METHODOLOGY/PRINCIPAL FINDINGS: Using a mouse model of oxygen-induced retinopathy, we found that while developmental angiogenesis is normal in PSMA null mice, hypoxic challenge resulted in decreased retinal vascular pathology when compared to wild type mice as assessed by avascular area and numbers of vascular tufts/glomeruli. The vessels formed in the PSMA null mice were more organized and highly perfused, suggesting a more 'normal' phenotype. Importantly, the decrease in angiogenesis was not due to an impaired hypoxic response as levels of pro-angiogenic factors are comparable; indicating that PSMA regulation of angiogenesis is independent of VEGF. Furthermore, both systemic and intravitreal administration of a PSMA inhibitor in wild type mice undergoing OIR mimicked the PSMA null phenotype resulting in improved retinal vasculature. CONCLUSIONS/SIGNIFICANCE: Our data indicate that PSMA plays a VEGF-independent role in retinal angiogenesis and that the lack of or inhibition of PSMA may

  17. FLUORESCENCE OVERLAY ANTIGEN MAPPING OF THE EPIDERMAL BASEMENT-MEMBRANE ZONE .1. GEOMETRIC ERRORS

    NARCIS (Netherlands)

    BRUINS, S; DEJONG, MCJM; HEERES, K; WILKINSON, MHF; JONKMAN, MF; VANDERMEER, JB

    1994-01-01

    To identify in tissue sections the relative positions of antigen distributions close to the resolving power of the microscope, we have developed the fluorescence overlay antigen mapping (FOAM) procedure. As this technique makes high demands on the geometric fidelity of the overlay image, it is essen

  18. Cdc42 and k-Ras Control Endothelial Tubulogenesis through Apical Membrane and Cytoskeletal Polarization: Novel Stimulatory Roles for GTPase Effectors, the Small GTPases, Rac2 and Rap1b, and Inhibitory Influence of Arhgap31 and Rasa1.

    Directory of Open Access Journals (Sweden)

    Pieter R Norden

    Full Text Available A critical and understudied property of endothelial cells is their ability to form lumens and tube networks. Although considerable information has been obtained concerning these issues, including the role of Cdc42 and Rac1 and their effectors such as Pak2, Pak4, Par6b, and co-regulators such as integrins, MT1-MMP and Par3; many key questions remain that are necessary to elucidate molecular and signaling requirements for this fundamental process. In this work, we identify new small GTPase regulators of EC tubulogenesis including k-Ras, Rac2 and Rap1b that act in conjunction with Cdc42 as well as the key downstream effectors, IQGAP1, MRCKβ, beta-Pix, GIT1, and Rasip1 (which can assemble into multiprotein complexes with key regulators including α2β1 integrin and MT1-MMP. In addition, we identify the negative regulators, Arhgap31 (by inactivating Cdc42 and Rac and Rasa1 (by inactivating k-Ras and the positive regulator, Arhgap29 (by inactivating RhoA which play a major functional role during the EC tubulogenic process. Human EC siRNA suppression or mouse knockout of Rasip1 leads to identical phenotypes where ECs form extensive cord networks, but cannot generate lumens or tubes. Essential roles for these molecules during EC tubulogenesis include; i establishment of asymmetric EC cytoskeletal polarization (subapical distribution of acetylated tubulin and basal membrane distribution of F-actin; and ii directed membrane trafficking of pinocytic vacuoles or other intracellular vesicles along acetylated tubulin tracks to the developing apical membrane surface. Cdc42 co-localizes subapically with acetylated tubulin, while Rac1 and k-Ras strongly label vacuole/ vesicle membranes which accumulate and fuse together in a polarized, perinuclear manner. We observe polarized apical membrane and subapical accumulation of key GTPases and effectors regulating EC lumen formation including Cdc42, Rac1, Rac2, k-Ras, Rap1b, activated c-Raf and Rasip1 to control EC

  19. The kinetics of antibody binding to Plasmodium falciparum VAR2CSA PfEMP1 antigen and modelling of PfEMP1 antigen packing on the membrane knobs

    DEFF Research Database (Denmark)

    Joergensen, Lars M; Salanti, Ali; Dobrilovic, Tina;

    2010-01-01

    ABSTRACT: BACKGROUND: Infected humans make protective antibody responses to the PfEMP1 adhesion antigens exported by Plasmodium falciparum parasites to the erythrocyte membrane, but little is known about the kinetics of this antibody-receptor binding reaction or how the topology of PfEMP1...... on the parasitized erythrocyte membrane influences antibody association with, and dissociation from, its antigenic target. METHODS: A Quartz Crystal Microbalance biosensor was used to measure the association and dissociation kinetics of VAR2CSA PfEMP1 binding to human monoclonal antibodies. Immuno....... RESULTS: Kinetic analysis indicates that antibody dissociation from the VAR2CSA PfEMP1 antigen is extremely slow when there is a high avidity interaction. High avidity binding to PfEMP1 antigens on the surface of P. falciparum-infected erythrocytes in turn requires bivalent cross-linking of epitopes...

  20. Definition of a physiologic aging autoantigen by using synthetic peptides of membrane protein band 3: localization of the active antigenic sites.

    Science.gov (United States)

    Kay, M M; Marchalonis, J J; Hughes, J; Watanabe, K; Schluter, S F

    1990-08-01

    Senescent cell antigen (SCA), an aging antigen, is a protein that appears on old cells and marks them for removal by the immune system in mammals. It is derived from band 3, a ubiquitous membrane transport protein found in diverse cell types and tissues. We have used synthetic peptides to identify aging antigenic sites on band 3, using a competitive inhibition assay and immunoblotting with IgG directed against the aging antigen on old cells. Results indicate that: (i) the active antigenic sites of the aging antigen reside on membrane protein band 3 residues that are extracellular regions implicated in anion transport (residues 538-554 and 788-827); (ii) a putative ankyrin-binding-region peptide is not involved in SCA activity; and (iii) carbohydrate moieties are not required for the antigenicity or recognition of SCA because synthetic peptides alone abolish binding of senescent cell IgG to erythrocytes. One of the putative transport sites that contributes to the aging antigen is located toward the carboxyl terminus. A model of band 3 is presented. Localization of the active antigenic site on the band 3 molecule facilitates definition of the molecular changes occurring during aging that initiate molecular as well as cellular degeneration. PMID:1696010

  1. Lysosome-associated membrane glycoprotein (LAMP)--preliminary study on a hidden antigen target for vaccination against schistosomiasis.

    Science.gov (United States)

    Nawaratna, Sujeevi S K; Gobert, Geoffrey N; Willis, Charlene; Mulvenna, Jason; Hofmann, Andreas; McManus, Donald P; Jones, Malcolm K

    2015-01-01

    Our previously reported gene atlasing of schistosome tissues revealed transcripts that were highly enriched in the digestive tract of Schistosoma mansoni. From these, we selected two candidates, Sm-LAMP and Sm-NPC2 for testing as vaccine targets. The two molecules were selected on the basis of relatively high expression in the gastrodermis, their potentially important biological function, divergence from homologous molecules of the host and possible apical membrane expression in the gastrodermis. Bacterially expressed recombinant peptides corresponding to regions excluding trans-membrane domains of the selected vaccine targets were used in blinded vaccine trials in CBA mice using alum-CpG as adjuvant. Vaccine trials using the recombinant insoluble Sm-LAMP protein showed 16-25% significant reduction in total worm burden. Faecal egg count reduction was 52% and 60% in two trials, respectively, with similar results for the solubly expressed protein. Liver egg burden was reduced significantly (20% and 38%) with an insoluble recombinant Sm-LAMP in two trials, but not with the soluble recombinant form. Parasite fecundity was not affected by either Sm-LAMP protein preparations in the trials. It is concluded that Sm-LAMP may provide limited protection towards S. mansoni infections but could be used in combination with other vaccine candidates, to provide more comprehensive protection. PMID:26472258

  2. A Novel Treponema pallidum Antigen, TP0136, Is an Outer Membrane Protein That Binds Human Fibronectin▿

    OpenAIRE

    Brinkman, Mary Beth; McGill, Melanie A.; Pettersson, Jonas; Rogers, Arthur; Matějková, Petra; Šmajs, David; Weinstock, George M.; Norris, Steven J; Palzkill, Timothy

    2008-01-01

    The antigenicity, structural location, and function of the predicted lipoprotein TP0136 of Treponema pallidum subsp. pallidum were investigated based on previous screening studies indicating that anti-TP0136 antibodies are present in the sera of syphilis patients and experimentally infected rabbits. Recombinant TP0136 (rTP0136) protein was purified and shown to be strongly antigenic during human and experimental rabbit infection. The TP0136 protein was exposed on the surface of the bacterial ...

  3. Metastatic superscan in prostate carcinoma on gallium-68-prostate-specific membrane antigen positron emission tomography/computed tomography scan

    Science.gov (United States)

    Agarwal, Krishan Kant; Tripathi, Madhavi; Kumar, Rajeev; Bal, Chandrasekhar

    2016-01-01

    We describe the imaging features of a metastatic superscan on gallium-68 Glu-NH-CO-NH-Lys-(Ahx)-[Ga-68(HBED-CC)], abbreviated as gallium-68-prostate-specific membrane antigen (68Ga-PSMA) positron emission tomography/computed tomography (PET/CT) imaging. 68Ga-PSMA is novel radiotracer undergoing evaluation for PET/CT imaging of prostate carcinoma. This patient had a superscan of metastases on conventional bone scintigraphy and was referred for 68Ga-PSMA PET/CT to evaluate the feasibility of 177Lu-PSMA therapy. PMID:27095868

  4. Prostate-specific membrane antigen expression in tumor-associated vasculature of breast cancers.

    Science.gov (United States)

    Wernicke, Alla Gabriella; Varma, Sonal; Greenwood, Eleni A; Christos, Paul J; Chao, K S Clifford; Liu, He; Bander, Neil H; Shin, Sandra J

    2014-06-01

    Prostate-specific membrane antigen (PSMA) has been found to be expressed in the tumor-associated neovasculature of multiple solid tumor types including breast cancers. However, thus far, the number of cases studied from some tumor types has been limited. In this study, we set out to assess PSMA expression in the tumor-associated vasculature associated with invasive breast carcinomas in a sizable cohort of patients. One hundred and six patients with AJCC stage 0-IV breast cancer were identified. Ninety-two of these patients had primary breast cancer [invasive breast carcinoma with or without co-existing ductal carcinoma in situ (DCIS) (74) or DCIS alone (18)]. In addition, 14 patients with breast cancer metastases to the brain were identified. Immunohistochemical staining for PSMA and CD31 was performed on parallel representative tumor sections in each case. Tumor-associated vascular endothelial cell PSMA immunoreactivity was semi-quantitatively assessed based on two parameters: overall percent of endothelial positivity and staining intensity. PSMA expression for tumor-associated vascular endothelial cells was scored 0 if there was no detectable PSMA expression, 1 if PSMA staining was detectable in 5-50%, and 2 if PSMA expression was positive in >50% of microvessels. CD 31 staining was concurrently reviewed to confirm the presence of vasculature in each case. Tumor-associated vasculature was PSMA-positive in 68/92 (74%) of primary breast cancers and in 14/14 (100%) of breast cancers metastatic to brain. PSMA was not detected in normal breast tissue or carcinoma cells. All but 2 cases (98%) showed absence of PSMA expression in normal breast tissue-associated vasculature. The 10-year overall survival was 88.7% (95% CI = 80.0%, 93.8%) in patients without brain metastases. When overall survival (OS) was stratified based on PSMA score group, patients with PSMA scores of 0, 1, and 2 had 10-year OS of 95.8%, 96.0%, and 79.7%, respectively (p = 0.12). When PSMA scores

  5. Potential Impact of Seasonal Malaria Chemoprevention on the Acquisition of Antibodies Against Glutamate-Rich Protein and Apical Membrane Antigen 1 in Children Living in Southern Senegal

    DEFF Research Database (Denmark)

    Ndiaye, Magatte; Sylla, Khadime; Sow, Doudou;

    2015-01-01

    Seasonal malaria chemoprevention (SMC) is defined as the intermittent administration of full treatment courses of an antimalarial drug to children during the peak of malaria transmission season with the aim of preventing malaria-associated mortality and morbidity. SMC using sulfadoxine-pyrimetham...

  6. Plasma membrane-bound AGC3 kinases phosphorylate PIN auxin carriers at TPRXS(N/S) motifs to direct apical PIN recycling

    NARCIS (Netherlands)

    P. Dhonukshe; F. Huang; C.S. Galvan-Ampudia; A.P. Mähönen; J. Kleine-Vehn; J. Xu; A. Quint; K. Prasad; J. Friml; B. Scheres; R. Offringa

    2010-01-01

    Polar membrane cargo delivery is crucial for establishing cell polarity and for directional transport processes. In plants, polar trafficking mediates the dynamic asymmetric distribution of PIN FORMED (PIN) carriers, which drive polar cell-to-cell transport of the hormone auxin, thereby generating a

  7. Targeting tumor antigens to secreted membrane vesicles in vivo induces efficient antitumor immune responses.

    NARCIS (Netherlands)

    Zeelenberg, I.S.; Ostrowski, M.; Krumeich, S.; Bobrie, A.; Jancic, C.; Boissonnas, A.; Delcayre, A.; Pecq, JB Le; Combadiere, B.; Amigorena, S.; Thery, C.

    2008-01-01

    Expression of non-self antigens by tumors can induce activation of T cells in vivo, although this activation can lead to either immunity or tolerance. CD8+ T-cell activation can be direct (if the tumor expresses MHC class I molecules) or indirect (after the capture and cross-presentation of tumor an

  8. Apical localization of PMCA2w/b is enhanced in terminally polarized MDCK cells

    OpenAIRE

    Antalffy, Géza; Caride, Ariel J.; Pászty, Katalin; Hegedus, Luca; Padanyi, Rita; STREHLER, EMANUEL E.; Enyedi, Ágnes

    2011-01-01

    The “w” splice forms of PMCA2 localize to distinct membrane compartments such as the apical membrane of the lactating mammary epithelium, the stereocilia of inner ear hair cells or the post-synaptic density of hippocampal neurons. Previous studies indicated that PMCA2w/b was not fully targeted to the apical domain of MDCK cells but distributed more evenly to the lateral and apical membrane compartments. Overexpression of the apical scaffold protein NHERF2, however, greatly increased the amoun...

  9. Ultrastructural demonstration of spirochetal antigens in synovial fluid and synovial membrane in chronic Lyme disease: possible factors contributing to persistence of organisms.

    Science.gov (United States)

    Nanagara, R; Duray, P H; Schumacher, H R

    1996-10-01

    To perform the first systematic electronmicroscopic (EM) and immunoelectron microscopy (IEM) study of the pathological changes and the evidence of spirochete presence in synovial membranes and synovial fluid (SF) cells of patients with chronic Lyme arthritis. EM examination was performed on four synovial membrane and eight SF cell samples from eight patients with chronic Lyme disease. Spirochetal antigens in the samples were sought by IEM using monoclonal antibody to Borrelia burgdorferi outer surface protein A (OspA) as the immunoprobe. Prominent ultrastructural findings were surface fibrin-like material, thickened synovial lining cell layer and signs of vascular injury. Borrelia-like structures were identified in all four synovial membranes and in two of eight SF cell samples. The presence of spirochetal antigens was confirmed by IEM in all four samples studied (one synovial membrane and three SF cell samples). OspA labelling was in perivascular areas, deep synovial stroma among collagen bundles, and in vacuoles of fibroblasts in synovial membranes; and in cytophagosomes of mononuclear cells in SF cell samples. Electron microscopy adds further evidence for persistence of spirochetal antigens in the joint in chronic Lyme disease. Locations of spirochetes or spirochetal antigens both intracellulary and extracellulary in deep synovial connective tissue as reported here suggest sites at which spirochaetes may elude host immune response and antibiotic treatment.

  10. Surface membrane antigen expression changes induced in vitro by exogenous growth factors in chronic lymphocytic leukemia cells.

    Science.gov (United States)

    Vilpo, J; Hulkkonen, J; Hurme, M; Vilpo, L

    2002-09-01

    The factors determining the growth and survival of cells in B chronic lymphocytic leukemia (CLL) have remained poorly understood. We investigated the effects of optimal mitogen combinations (OMCs) on the expression of 26 surface membrane antigens among 33 CLL patients. The seven OMCs used were selected after pre-testing 14 combinations of (1) S. aureus Cowan I (SAC), (2) interleukin-2 (IL-2), (3) tumor necrosis factor alpha (TNF-alpha) and (4) 12-O-tetradecanoylphorbol 13-acetate (TPA; also known as phorbol 12-myristate 13-acetate or PMA). In flow cytometry we revealed that OMCs induced statistically highly significant upregulation of the expression of CD5, CD11c, CD19, CD22, CD23, CD25, CD38, CD40, CD45, CD45RO, CD95, CD126, CD130 and FMC7, and downregulation of CD20 and CD124 expression. Interestingly, the expression of CD27, CD45RA, CD79b, CD80, CD122 and that of the immunoglobulin gene superfamily members CD21, Ig-kappa, Ig-lambda, Ig-delta and Ig-micro were not significantly affected under similar conditions. The expression of several antigens was co-regulated, suggesting common regulatory pathways. These antigens include CD11c/CD5, CD11c/CD22, CD11c/CD126, CD11c/FMC7 as well as CD27/CD45, CD27/CD45RA and CD27/CD79b. Upregulation of surface antigen expression, induced by OMCs, should be applicable in antibody therapy in vitro and in vivo, and in negative stem cell selection for autotransplantation. Furthermore, the current strategy to enhance cell surface antigen expression may be a versatile tool to raise humoral and cell-mediated host defense against CLL cells. Upregulation of proteins mediating positive growth signals (eg CD25, CD40) and negative signals or apoptosis (eg CD95) may be used to sensitize cells to chemotherapy and programmed cell death. PMID:12200683

  11. Plasma membrane associated, virus-specific polypeptides required for the formation of target antigen complexes recognized by virus-specific cytotoxic T lymphocytes

    International Nuclear Information System (INIS)

    These studies were undertaken to define some of the poxvirus-specific target antigens which are synthesized in infected cells and recognized by vaccinia virus-specific CTLs (VV-CTLs). Since vaccinia virus infected, unmanipulated target cells express numerous virus-specific antigens on the plasma membrane, attempts were made to manipulate expression of the poxvirus genome after infection so that one or a few defined virus-specified antigens were expressed on the surface of infected cells. In vitro [51Cr]-release assays determined that viral DNA synthesis and expression of late viral proteins were not necessary to form a target cell which was fully competent for lysis by VV-CTLs. Under the conditions employed in these experiments, 90-120 minutes of viral protein synthesis were necessary to produce a competent cell for lysis by VV-CTLs. In order to further inhibit the expression of early viral proteins in infected cells, partially UV-inactivated vaccinia virus was employed to infect target cells. It was determined that L-cells infected with virus preparations which had been UV-irradiated for 90 seconds were fully competent for lysis by VV-CTLs. Cells infected with 90 second UV-irr virus expressed 3 predominant, plasma membrane associated antigens of 36-37K, 27-28K, and 19-17K. These 3 viral antigens represent the predominant membrane-associated viral antigens available for interaction with class I, major histocompatibility antigens and hence are potential target antigens for VV-CTLs

  12. Proteomic analysis of liver plasma membrane from hepatitis B surface antigen transgenic mice

    Institute of Scientific and Technical Information of China (English)

    贾小芳

    2012-01-01

    Objective To explore the differential liver plasma membrane( PM) proteins that may be related to the occurrence,development and reversal process of hepatitis and to understand the pathogenesis of hepatitis and the new drug targets by performing a comparative proteomics research of liver PM between

  13. Deposition of nucleosomal antigens (histones and DNA) in the epidermal basement membrane in human lupus nephritis.

    NARCIS (Netherlands)

    Grootscholten, C.; Bruggen, M.C.J. van; Pijl, J.W. van der; Jong, E.M.G.J. de; Ligtenberg, G.; Derksen, R.H.W.M.; Berden, J.H.M.

    2003-01-01

    OBJECTIVE: Antinuclear autoantibodies complexed to nucleosomes can bind to heparan sulfate (HS) in the glomerular basement membrane. This binding is due to the binding of the positively charged histones to the strongly anionic HS. Nucleosomes and histones have been identified in glomerular deposits

  14. Targeting LSCs through membrane antigens selectively or preferentially expressed on these cells.

    Science.gov (United States)

    Pelosi, Elvira; Castelli, Germana; Testa, Ugo

    2015-12-01

    Studies of xenotransplantation of bone marrow and blood cells of AML patients have supported the existence of rare leukemic stem cells, able to initiate and maintain the leukemic process and bearing the typical leukemic abnormalities. LSCs possess self-renewal capacity and are responsible for the growth of the more differentiated leukemic progeny in the bone marrow and in the blood. These cells are more resistant than bulk leukemic cells to anti-leukemic drugs, thus survive to treatment and are, at a large extent, responsible for leukemia relapse. During the last two decades, considerable progresses have been made in the understanding of the peculiar cellular and molecular properties of LSCs. In this context, particularly relevant was the discovery of several membrane markers, selectively or preferentially expressed on LSCs. These membrane markers offer now unique opportunities to identify LSCs and to distinguish them from normal HSCs, to monitor the response of the various anti-leukemic treatments at the level of the LSC compartment, to identify relevant therapeutic targets. Concerning this last point, the most promising therapeutic targets are CD33 and CD123.

  15. Membrane and envelope virus proteins co-expressed as lysosome associated membrane protein (LAMP fused antigens: a potential tool to develop DNA vaccines against flaviviruses

    Directory of Open Access Journals (Sweden)

    Rafael Dhalia

    2009-12-01

    Full Text Available Vaccination is the most practical and cost-effective strategy to prevent the majority of the flavivirus infection to which there is an available vaccine. However, vaccines based on attenuated virus can potentially promote collateral side effects and even rare fatal reactions. Given this scenario, the developent of alternative vaccination strategies such as DNA-based vaccines encoding specific flavivirus sequences are being considered. Endogenous cytoplasmic antigens, characteristically plasmid DNA-vaccine encoded, are mainly presented to the immune system through Major Histocompatibility Complex class I - MHC I molecules. The MHC I presentation via is mostly associated with a cellular cytotoxic response and often do not elicit a satisfactory humoral response. One of the main strategies to target DNA-encoded antigens to the MHC II compartment is expressing the antigen within the Lysosome-Associated Membrane Protein (LAMP. The flavivirus envelope protein is recognized as the major virus surface protein and the main target for neutralizing antibodies. Different groups have demonstrated that co-expression of flavivirus membrane and envelope proteins in mammalian cells, fused with the carboxyl-terminal of LAMP, is able to induce satisfactory levels of neutralizing antibodies. Here we reviewed the use of the envelope flavivirus protein co-expression strategy as LAMP chimeras with the aim of developing DNA vaccines for dengue, West Nile and yellow fever viruses.A vacinação é a estratégia mais prática e o melhor custo-benefício para prevenir a maioria das infecções dos flavivirus, para os quais existe vacina disponível. Entretanto, as vacinas baseadas em vírus atenuados podem potencialmente promover efeitos colaterais e, mais raramente, reações fatais. Diante deste cenário, o desenvolvimento de estratégias alternativas de vacinação, como vacinas baseadas em DNA codificando seqüências específicas dos flavivirus, está sendo considerado

  16. A restricted set of apical proteins recycle through the trans-Golgi network in MDCK cells.

    OpenAIRE

    Brändli, A W; Simons, K

    1989-01-01

    Sorting of newly synthesized proteins destined for the apical plasma membrane takes place in the trans-Golgi network (TGN) in MDCK cells. This process is most likely receptor mediated and requires components that recycle between both compartments. We have developed an assay to detect apical proteins that recycle through the sialyltransferase-containing TGN. Cell surface glycoproteins were exogalactosylated apically using a mutant cell line derived from MDCK, MDCKII-RCAr. The mutant exhibits i...

  17. Multiple components of blood group A and B antigens in human erythrocyte membranes and their difference between A1 and A2 status.

    OpenAIRE

    Fujii, H.; Yoshida, A.

    1980-01-01

    Human type O erythrocyte membranes were converted to type A1 by purified human A1-enzyme, to type A2 by purified human A2-enzyme, and to type B by purified human B-enzyme in the presence of radioactive sugar donors (i.e., UDP-N-acetylgalactosamine for A-enzyme and UDP-galactose for B-enzyme, respectively). Type A2 erythrocyte membranes were also converted to type A1 by purified A1-enzyme A1-enzyme. The labeled blood group antigens (A1, A2, and B) thus produced were analyzed by sodium dodecyl ...

  18. Design and preclinical evaluation of a 99mTc-labelled diabody of mAb J591 for SPECT imaging of prostate-specific membrane antigen (PSMA)

    OpenAIRE

    Kampmeier, Florian; Williams, Jennifer D.; Maher, John; Mullen, Gregory E.; Blower, Philip J.

    2014-01-01

    Background Sensitive and specific detection of nodal status, sites of metastases and low-volume recurrent disease could greatly improve management of patients with advanced prostate cancer. Prostate-specific membrane antigen (PSMA) is a well-established marker for prostate carcinoma with increased levels of expression in high-grade, hormone-refractory and metastatic disease. The monoclonal antibody (mAb) J591 is directed against an extracellular epitope of PSMA and has been shown to efficient...

  19. Oral Immunization with Recombinant Mycobacterium smegmatis Expressing the Outer Membrane Protein 26-Kilodalton Antigen Confers Prophylactic Protection against Helicobacter pylori Infection ▿ †

    OpenAIRE

    Lü, Lin; Zeng, Han-qing; Wang, Pi-Long; Shen, Wei; Xiang, Ting-xiu; Mei, Zhe-chuan

    2011-01-01

    Helicobacter pylori infection is prevalent worldwide and results in chronic gastritis, which may lead to gastric mucosa-associated lymphoid tissue lymphoma and gastric cancer. We have previously reported that oral immunization with recombinant Mycobacterium smegmatis expressing the H. pylori outer membrane protein 26-kilodalton (Omp26) antigen affords therapeutic protection against H. pylori infection in mice. In the present study, we investigated the prophylactic effects of this vaccine cand...

  20. Determination of the genus-specific antigens in outer membrane proteins from the strains of Leptospira interrogans and Leptospira biflexa with different virulence

    Institute of Scientific and Technical Information of China (English)

    罗依惠; 严杰; 毛亚飞; 李淑萍

    2004-01-01

    Objective:To determine the existence of genus-specific antigens in outer membrane proteins (OMPs) of leptospira with different virulence. Methods: Microscope agglutination test (MAT) was applied to detect the agglutination between commercial rabbit antiserum against leptospiral genus-specific TR/Patoc I antigen and 17 strains of Leptospira interrongans belonging to 15 serogroups and 2 strains of Leptospira biflexa belonging to 2 serogroups.The outer envelopes (OEs) of L.interrogans serogroup Icterohaemorrhagiae serovar lai strain lai (56601) with strong virulence and serogroup Pomona serovar pomona strain Luo (56608) with low virulence,and L.biflexa serogroup Semaranga serovar patoc strain Patoc I without virulence were prepared by using the method reported in Auran et al.(1972).OMPs in the OEs were obtained by treatment with sodium deoxycholate. SDS-PAGE and western blot were used for analyzing the features of the OMPs on electrophoretic pattern and the immunoreactivity to the antiserum against TR/Patoc I antigen, respectively. Results:All the tested strains belonging to different leptospiral serogroups agglutinated to the antiserum against leptospiral genus-specific TR/Patoc I antigen with agglutination titers ranging from 1:256-1:512. A similar SDS-PAGE pattern of the OMPs from the three strains of leptospira with different virulence was shown and the molecular weight of a major protein fragment in the OMPs was found to be approximately 60 KDa.A positive protein fragment with approximately 32 KDa confirmed by Western blot,was able to react with the antiserum against leptospiral genus-specific TR/Patoc I antigen, and was found in each the OMPs of the three stains of leptospira.Conclusion: There are genus-specific antigens on the surface of L.interrogans and L.biflexa. The OMP with molecular weight of 32 KDa may be one of the genus-specific protein antigens of leptospira.

  1. Determination of the genus-specific antigens in outer membrane proteins from the strains of Leptospira interrogans and Leptospira biflexa with different virulence

    Institute of Scientific and Technical Information of China (English)

    罗依惠; 严杰; 毛亚飞; 李淑萍

    2004-01-01

    Objective: To determine the existence of genus-specific antigens in outer membrane proteins (OMPs) of leptospira with different virulence. Methods: Microscope agglutination test (MAT) was applied to detect the agglutination between commercial rabbit antiserum against leptospiral genus-specific TR/Patoc I antigen and 17 strains of Leptospira interrongans belonging to 15 serogroups and 2 strains of Leptospira biflexa belonging to 2 serogroups. The outer envelopes (OEs) of L.interrogans serogroup Icterohaemorrhagiae serovar lai strain lai (56601) with strong virulence and serogroup Pomona serovar pomona strain Luo (56608) with low virulence, and L.biflexa serogroup Semaranga serovar patoc strain Patoc I without virulence were prepared by using the method reported in Auran et al.(1972). OMPs in the OEs were obtained by treatment with sodium deoxycholate. SDS-PAGE and western blot were used for analyzing the features of the OMPs on electrophoretic pattern and the immunoreactivity to the antiserum against TR/Patoc I antigen, respectively. Results: All the tested strains belonging to different leptospiral serogroups agglutinated to the antiserum against leptospiral genus-specific TR/Patoc I antigen with agglutination titers ranging from 1:256-1:512. A similar SDS-PAGE pattern of the OMPs from the three strains of leptospira with different virulence was shown and the molecular weight of a major protein fragment in the OMPs was found to be approximately 60 KDa. A positive protein fragment with approximately 32 KDa confirmed by Western blot, was able to react with the antiserum against leptospiral genus-specific TR/Patoc I antigen, and was found in each the OMPs of the three stains of leptospira. Conclusion: There are genus-specific antigens on the surface of L.interrogans and L.biflexa. The OMP with molecular weight of 32 KDa may be one of the genus-specific protein antigens of leptospira.

  2. Functional Recombinant Extra Membrane Loop of Human CD20, an Alternative of the Full Length CD20 Antigen

    OpenAIRE

    Anbouhi, Mahdi Habibi; Baraz, Aida Feiz; Bouzari, Saeid; Abolhassani, Mohsen; Khanahmad, Hossein; Golkar, Majid; Aghasadeghi, Mohammad Reza; Behdani, Mahdi; Najafabadi, Ali Jahanian; Shokrgozar, Mohammad Ali

    2012-01-01

    Background: Targeting of CD20 antigen with monoclonal antibodies has become the mainstay in the treatment of non-Hodgkin's lymphomas and immunotherapeutic depletion of malignant B cells. Accessibility of antigen is one of the crucial factors in development of monoclonal antibodies against this antigen. One major problem in expression of full length CD20 is aggregation and misfolding. Therefore, production of an alternative polypeptide is easer and favorable comparing to that of a full length ...

  3. Immunogenicity and protective role of antigenic regions from five outer membrane proteins of Flavobacterium columnare in grass carp Ctenopharyngodon idella

    Science.gov (United States)

    Luo, Zhang; Liu, Zhixin; Fu, Jianping; Zhang, Qiusheng; Huang, Bei; Nie, Pin

    2016-02-01

    Flavobacterium columnare causes columnaris disease in freshwater fish. In the present study, the antigenic regions of five outer membrane proteins (OMPs), including zinc metalloprotease, prolyl oligopeptidase, thermolysin, collagenase and chondroitin AC lyase, were bioinformatically analyzed, fused together, and then expressed as a recombinant fusion protein in Escherichia coli. The expressed protein of 95.6 kDa, as estimated by 10% sodium dodecyl sulfate-polyacrylamide gel electrophoresis, was consistent with the molecular weight deduced from the amino acid sequence. The purified recombinant protein was used to vaccinate the grass carp, Ctenopharyngodon idella. Following vaccination of the fish their IgM antibody levels were examined, as was the expression of IgM, IgD and IgZ immunoglobulin genes and other genes such as MHC Iα and MHC IIβ, which are also involved in adaptive immunity. Interleukin genes (IL), including IL-1β, IL-8 and IL-10, and type I and type II interferon (IFN) genes were also examined. At 3 and 4 weeks post-vaccination (wpv), significant increases in IgM antibody levels were observed in the fish vaccinated with the recombinant fusion protein, and an increase in the expression levels of IgM, IgD and IgZ genes was also detected following the vaccinations, thus indicating that an adaptive immune response was induced by the vaccinations. Early increases in the expression levels of IL and IFN genes were also observed in the vaccinated fish. At four wpv, the fish were challenged with F. columnare, and the vaccinated fish showed a good level of protection against this pathogen, with 39% relative percent survival (RPS) compared with the control group. It can be concluded, therefore, that the five OMPs, in the form of a recombinant fusion protein vaccine, induced an immune response in fish and protection against F. columnare.

  4. Expression of prostate-specific membrane antigen in lung cancer cells and tumor neovasculature endothelial cells and its clinical significance.

    Directory of Open Access Journals (Sweden)

    Hai-long Wang

    Full Text Available Prostate-specific membrane antigen (PSMA has been found in tumor neovasculature endothelial cells (NECs of non-prostate cancers and may become the most promising target for anti-tumor therapy. To study the value of PSMA as a potential new target for lung cancer treatment, PSMA expression in non-small cell lung cancer (NSCLC and small cell lung cancer (SCLC tissues and its relationship with clinicopathology were investigated in the current study.Immunohistochemistry was used to detect PSMA expression in a total of 150 lung specimens of patients with lung cancer. The data were analyzed using univariate and multivariate statistical analyses.The percentages of NSCLC patients who had PSMA (+ tumor cells and PSMA (+ NECs were 54.02% and 85.06%, respectively. The percentage of patients younger than 60 years old who had PSMA (+ tumor cells was 69.05%, which was significantly greater than the percentage of patients aged 60 years or older (40.00%, p<0.05. A significant difference was observed in the percentage of NSCLC patients with PMSA (+ NECs and stage I or II cancer (92.98% and those patients with stage III or IV cancer (76.77%. In the SCLC tissues, NEC PSMA expression (70.00% did not differ significantly from NSCLC. SCLC tumor cells and normal lung tissues cells were all negative. There was no significant correlation between the presence of PSMA (+ NECs in SCLC patients and the observed clinicopathological parameters.PSMA is expressed not only in NECs of NSCLC and SCLC but also in tumor cells of most NSCLC patients. The presence of PSMA (+ tumor cells and PSMA (+ NECs in NSCLC was negatively correlated with age and the clinicopathological stage of the patients, respectively.

  5. Roles of external and cellular Cl- ions on the activation of an apical electrodiffusional Cl- pathway in toad skin.

    Science.gov (United States)

    Procopio, J; Lacaz-Vieira, F

    1990-07-01

    This study is concerned with the short-circuit current, Isc, responses of the Cl(-)-transporting cells of toad skin submitted to sudden changes of the external Cl- concentration, [Cl]o. Sudden changes of [Cl]o, carried out under apical membrane depolarization, allowed comparison of the roles of [Cl]o and [Cl]cell on the activation of the apical Cl- pathways. Equilibration of short-circuited skins symmetrically in K-Ringer's solutions of different Cl- concentrations permitted adjustment of [Cl]cell to different levels. For a given Cl- concentration (in the range of 11.7 to 117 mM) on both sides of a depolarized apical membrane, this structure exhibits a high Cl- permeability, P(Cl)apical. On the other hand, for the same range of [Cl]cell but with [Cl]o = 0, P(Cl)apical is reduced to negligible values. These observations indicate that when the apical membrane is depolarized P(Cl)apical is modulated by [Cl]o; in the absence of external Cl- ions, intracellular Cl- is not sufficient to activate P(Cl)apical. Computer simulation shows that the fast Cl- currents induced across the apical membrane by sudden shifts of [Cl]o from a control equilibrium value strictly follow the laws of electrodiffusion. For each experimental group, the computer-generated Isc versus [( Cl]cell - [Cl]o) curve which best fits the experimental data can only be obtained by a unique pair of P(Cl)apical and Rb (resistance of the basolateral membrane), thus allowing the calculation of these parameters. The electrodiffusional behavior of the net Cl- flux across the apical membrane supports the channel nature of the apical Cl- pathways in the Cl(-)-transporting cells. Cl- ions contribute significantly to the overall conductance of the basolateral membrane even in the presence of a high K concentration in the internal solution. PMID:1698229

  6. Monoclonal Antibodies as Probes for Unique Antigens in Secretory Cells of Mixed Exocrine Organs

    Science.gov (United States)

    Basbaum, C. B.; Mann, J. K.; Chow, A. W.; Finkbeiner, W. E.

    1984-07-01

    In the past, it has been difficult to identify the secretory product and control mechanisms associated with individual cell types making up mixed exocrine organs. This report establishes the feasibility of using immunological methods to characterize both the biochemical constituents and regulatory mechanisms associated with secretory cells in the trachea. Monoclonal antibodies directed against components of tracheal mucus were produced by immunizing mice with dialyzed, desiccated secretions harvested from tracheal organ culture. An immunofluorescence assay revealed that of the total 337 hybridomas screened, 100 produced antibodies recognizing goblet cell granules; 64, gland cell granules; and 3, antigen confined to the ciliated apical surface of the epithelium. The tracheal goblet cell antibody described in this report was strongly cross-reactive with intestinal goblet cells, as well as with a subpopulation of submandibular gland cells, but not with cells of Brunner's glands or the ciliated cell apical membrane. The serous cell antibody was not cross-reactive with goblet, Brunner's gland, or submandibular cells, or the ciliated cell apical membrane. The antibody directed against the apical membrane of ciliated cells did not cross-react with gland or goblet cells or the apical membrane of epithelial cells in the duodenum. Monoclonal antibodies, therefore, represent probes by which products unique to specific cells or parts of cells in the trachea can be distinguished. The antibodies, when used in enzyme immunoassays, can be used to quantitatively monitor secretion by individual cell types under a variety of physiological and pathological conditions. They also provide the means for purification and characterization of cell-specific products by immunoaffinity chromatography.

  7. Prostate-specific membrane antigen targeted protein contrast agents for molecular imaging of prostate cancer by MRI

    Science.gov (United States)

    Pu, Fan; Salarian, Mani; Xue, Shenghui; Qiao, Jingjuan; Feng, Jie; Tan, Shanshan; Patel, Anvi; Li, Xin; Mamouni, Kenza; Hekmatyar, Khan; Zou, Juan; Wu, Daqing; Yang, Jenny J.

    2016-06-01

    Prostate-specific membrane antigen (PSMA) is one of the most specific cell surface markers for prostate cancer diagnosis and targeted treatment. However, achieving molecular imaging using non-invasive MRI with high resolution has yet to be achieved due to the lack of contrast agents with significantly improved relaxivity for sensitivity, targeting capabilities and metal selectivity. We have previously reported our creation of a novel class of protein Gd3+ contrast agents, ProCA32, which displayed significantly improved relaxivity while exhibiting strong Gd3+ binding selectivity over physiological metal ions. In this study, we report our effort in further developing biomarker-targeted protein MRI contrast agents for molecular imaging of PSMA. Among three PSMA targeted contrast agents engineered with addition of different molecular recognition sequences, ProCA32.PSMA exhibits a binding affinity of 1.1 +/- 0.1 μM for PSMA while the metal binding affinity is maintained at 0.9 +/- 0.1 × 10-22 M. In addition, ProCA32.PSMA exhibits r1 of 27.6 mM-1 s-1 and r2 of 37.9 mM-1 s-1 per Gd (55.2 and 75.8 mM-1 s-1 per molecule r1 and r2, respectively) at 1.4 T. At 7 T, ProCA32.PSMA also has r2 of 94.0 mM-1 s-1 per Gd (188.0 mM-1 s-1 per molecule) and r1 of 18.6 mM-1 s-1 per Gd (37.2 mM-1 s-1 per molecule). This contrast capability enables the first MRI enhancement dependent on PSMA expression levels in tumor bearing mice using both T1 and T2-weighted MRI at 7 T. Further development of these PSMA-targeted contrast agents are expected to be used for the precision imaging of prostate cancer at an early stage and to monitor disease progression and staging, as well as determine the effect of therapeutic treatment by non-invasive evaluation of the PSMA level using MRI.Prostate-specific membrane antigen (PSMA) is one of the most specific cell surface markers for prostate cancer diagnosis and targeted treatment. However, achieving molecular imaging using non-invasive MRI with high

  8. [18F]Fluorobenzoyllysinepentanedioic Acid Carbamates: New Scaffolds for Positron Emission Tomography (PET) Imaging of Prostate-Specific Membrane Antigen (PSMA)

    OpenAIRE

    Yang, Xing; Mease, Ronnie C.; Pullambhatla, Mrudula; Lisok, Ala; Chen, Ying; Foss, Catherine A.; Wang, Yuchuan; Shallal, Hassan; Edelman, Hannah; Hoye, Adam T.; Attardo, Giorgio; Nimmagadda, Sridhar; Pomper, Martin G.

    2015-01-01

    Radiolabeled urea-based low-molecular weight inhibitors of the prostate-specific membrane antigen (PSMA) are under intense investigation as imaging and therapeutic agents for prostate and other cancers. In an effort to provide agents with less nontarget organ uptake than the ureas, we synthesized four 18F-labeled inhibitors of PSMA based on carbamate scaffolds. 4-Bromo-2-[18F]fluorobenzoyllysineoxypentanedioic acid (OPA) carbamate [18F]23 and 4-iodo-2-[18F]fluorobenzoyllysine OPA carbamate [1...

  9. Proteomic study via a non-gel based approach of meningococcal outer membrane vesicle vaccine obtained from strain CU385: a road map for discovering new antigens.

    Science.gov (United States)

    Gil, Jeovanis; Betancourt, L Zaro H; Sardiñas, Gretel; Yero, Daniel; Niebla, Olivia; Delgado, Maité; García, Darien; Pajón, Rolando; Sánchez, Aniel; González, Luis J; Padrón, Gabriel; Campa, Concepción; Sotolongo, Franklin; Barberó, Ramón; Guillén, Gerardo; Herrera, Luis; Besada, Vladimir

    2009-05-01

    This work presents the results from a study of the protein composition of outer membrane vesicles from VA-MENGOC-BC (Finlay Institute, Cuba), an available vaccine against serogroup B Neisseria meningitidis. Proteins were identified by means of SCAPE, a 2DE-free method for proteome studies. More than one hundred proteins were detected by tandem liquid chromatographymass spectrometry analysis of fractions enriched in peptides devoid of histidine or arginine residues, providing a detailed description of the vaccine. A bioinformatic analysis of the identified components resulted in the identification of 31 outer membrane proteins and three conserved hypothetical proteins, allowing the cloning, expression, purification and immunological study of two of them (NMB0088 and NMB1796) as new antigens.

  10. Increased basolateral sorting of carcinoembryonic antigen in a polarized colon carcinoma cell line after cholesterol depletion-Implications for treatment of inflammatory bowel disease

    Institute of Scientific and Technical Information of China (English)

    Robert Ehehalt; Markus Krautter; Martin Zorn; Richard Sparla; Joachim Fūllekrug; Hasan Kulaksiz; Wolfgang Stremmel

    2008-01-01

    AIM:To investigate a possible increase of basolateral expression of carcinoembryonic antigen(CEA)by interfering with the apical transport machinery,we studied the effect of cholesterol depletion on CEA sorting and secretion.METHODS:Cholesterol depletion was performed in polarized Caco-2 cells using Iovastatin and methyl-βcyclodextrin.RESULTS:We show that CEA is predominantly expressed and secreted at the apical surface.Reduction of the cholesterol level of the cell by 40%-50% with Iovastatin and methyl-β-cyclodextrin led to a significant change of the apical-to-basolateral transport ratio towards the basolateral membrane.CONCLUSION:As basolateral expression of CEA has been suggested to have anti-inflamatory properties,Cholesterol depletion of enterocytes might be a potential approach to influence the course of inflammatory bowel disease.

  11. The two neutrophil plasma membrane markers alkaline phosphatase and HLA class I antigen localize differently in granule-deficient cytoplasts. An ideal plasma membrane marker in human neutrophils is still lacking.

    Science.gov (United States)

    Pellmé, Sara; Dahlgren, Claes; Karlsson, Anna

    2007-08-31

    Neutrophil function relies largely on the ability of the cell to mobilize its different granules and vesicles to the cell surface and thereby expose and/or release effector molecules to the surrounding tissue. To properly identify these subcellular compartments is thus a prerequisite for studies of neutrophil physiology. A range of specific markers for the classical granules is available, but finding optimal markers for the secretory vesicles and plasma membrane has historically been more challenging. Latent and non-latent alkaline phosphatase activities are often used to distinguish these two light membrane structures, but the outcome using this technique depends on the level of cellular activation. Therefore, HLA-I was introduced some years ago as a specific, stimulation-independent marker for the plasma membrane. In this study we however report that detailed fractionation studies of neutrophil cytoplasts, lacking secretory vesicles, granules and other dense organelles, reveal that the HLA-I antigen is not only co-localizing with the plasma membrane marker ALP, but is also present in other, more dense organelles. Further, we found the mixed enzyme-linked immunosorbent assay (MELISA), detecting the beta(2)-microglobulin/HLA-I complex, to be negatively influenced by uncomplexed beta(2)-microglobulin present in the specific granules and secretory vesicles, making it difficult to use HLA-I as a plasma membrane marker during maturation of for example phagolysosomes. PMID:17673253

  12. Copper directs ATP7B to the apical domain of hepatic cells via basolateral endosomes.

    Science.gov (United States)

    Nyasae, Lydia K; Schell, Michael J; Hubbard, Ann L

    2014-12-01

    Physiologic Cu levels regulate the intracellular location of the Cu ATPase ATP7B. Here, we determined the routes of Cu-directed trafficking of endogenous ATP7B in the polarized hepatic cell line WIF-B and in the liver in vivo. Copper (10 µm) caused ATP7B to exit the trans-Golgi network (TGN) in vesicles, which trafficked via large basolateral endosomes to the apical domain within 1 h. Although perturbants of luminal acidification had little effect on the TGN localization of ATP7B in low Cu, they blocked delivery to the apical membrane in elevated Cu. If the vesicular proton-pump inhibitor bafilomycin-A1 (Baf) was present with Cu, ATP7B still exited the TGN, but accumulated in large endosomes located near the coverslip, in the basolateral region. Baf washout restored ATP7B trafficking to the apical domain. If ATP7B was staged apically in high Cu, Baf addition promoted the accumulation of ATP7B in subapical endosomes, indicating a blockade of apical recycling, with concomitant loss of ATP7B at the apical membrane. The retrograde pathway to the TGN, induced by Cu removal, was far less affected by Baf than the anterograde (Cu-stimulated) case. Overall, loss of acidification-impaired Cu-regulated trafficking of ATP7B at two main sites: (i) sorting and exit from large basolateral endosomes and (ii) recycling via endosomes near the apical membrane. PMID:25243755

  13. CD8+ T Cell Fate and Function Influenced by Antigen-Specific Virus-Like Nanoparticles Co-Expressing Membrane Tethered IL-2.

    Directory of Open Access Journals (Sweden)

    Daniela Wojta-Stremayr

    Full Text Available A variety of adjuvants fostering humoral immunity are known as of today. However, there is a lack of adjuvants or adjuvant strategies, which directly target T cellular effector functions and memory. We here determined whether systemically toxic cytokines such as IL-2 can be restricted to the site of antigen presentation and used as 'natural adjuvants'. Therefore, we devised antigen-presenting virus-like nanoparticles (VNP co-expressing IL-2 attached to different membrane-anchors and assessed their potency to modulate CD8+ T cell responses in vitro and in vivo. Efficient targeting of IL-2 to lipid rafts and ultimately VNP was achieved by fusing IL-2 at its C-terminus to a minimal glycosylphosphatidylinositol (GPI-anchor acceptor sequence. To identify optimal membrane-anchor dimensions we inserted one (1Ig, two (2Ig or four (4Ig immunoglobulin(Ig-like domains of CD16b between IL-2 and the minimal GPI-anchor acceptor sequence of CD16b (GPI. We found that the 2IgGPI version was superior to all other evaluated IL-2 variants (IL-2v in terms of its i degree of targeting to lipid rafts and to the VNP surface, ii biological activity, iii co-stimulation of cognate T cells in the absence of bystander activation and iv potency to induce differentiation and acquisition of CD8+ T cell effector functions in vitro and in vivo. In contrast, the GPI version rather favored memory precursor cell formation. These results exemplify novel beneficial features of membrane-bound IL-2, which in addition to its mere T cell stimulatory capacity include the induction of differential effector and memory functions in CD8+ T lymphocytes.

  14. Membrane and envelope virus proteins co-expressed as lysosome associated membrane protein (LAMP) fused antigens: a potential tool to develop DNA vaccines against flaviviruses

    OpenAIRE

    Rafael Dhalia; Milton Maciel Jr.; Fábia S.P. Cruz; Isabelle F.T. Viana; Mariana L. Palma; Thomas August; Ernesto T.A. Marques Jr.

    2009-01-01

    Vaccination is the most practical and cost-effective strategy to prevent the majority of the flavivirus infection to which there is an available vaccine. However, vaccines based on attenuated virus can potentially promote collateral side effects and even rare fatal reactions. Given this scenario, the developent of alternative vaccination strategies such as DNA-based vaccines encoding specific flavivirus sequences are being considered. Endogenous cytoplasmic antigens, characteristically plasmi...

  15. Membraner

    DEFF Research Database (Denmark)

    Bach, Finn

    2009-01-01

    Notatet giver en kort introduktion til den statiske virkemåde af membraner og membrankonstruktioner......Notatet giver en kort introduktion til den statiske virkemåde af membraner og membrankonstruktioner...

  16. Genetic mapping identifies novel highly protective antigens for an apicomplexan parasite.

    Directory of Open Access Journals (Sweden)

    Damer P Blake

    Full Text Available Apicomplexan parasites are responsible for a myriad of diseases in humans and livestock; yet despite intensive effort, development of effective sub-unit vaccines remains a long-term goal. Antigenic complexity and our inability to identify protective antigens from the pool that induce response are serious challenges in the development of new vaccines. Using a combination of parasite genetics and selective barriers with population-based genetic fingerprinting, we have identified that immunity against the most important apicomplexan parasite of livestock (Eimeria spp. was targeted against a few discrete regions of the genome. Herein we report the identification of six genomic regions and, within two of those loci, the identification of true protective antigens that confer immunity as sub-unit vaccines. The first of these is an Eimeria maxima homologue of apical membrane antigen-1 (AMA-1 and the second is a previously uncharacterised gene that we have termed 'immune mapped protein-1' (IMP-1. Significantly, homologues of the AMA-1 antigen are protective with a range of apicomplexan parasites including Plasmodium spp., which suggest that there may be some characteristic(s of protective antigens shared across this diverse group of parasites. Interestingly, homologues of the IMP-1 antigen, which is protective against E. maxima infection, can be identified in Toxoplasma gondii and Neospora caninum. Overall, this study documents the discovery of novel protective antigens using a population-based genetic mapping approach allied with a protection-based screen of candidate genes. The identification of AMA-1 and IMP-1 represents a substantial step towards development of an effective anti-eimerian sub-unit vaccine and raises the possibility of identification of novel antigens for other apicomplexan parasites. Moreover, validation of the parasite genetics approach to identify effective antigens supports its adoption in other parasite systems where legitimate

  17. Apical ammonium inhibition of cAMP-stimulated secretion in T84 cells is bicarbonate dependent.

    Science.gov (United States)

    Worrell, Roger T; Best, Alison; Crawford, Oscar R; Xu, Jie; Soleimani, Manoocher; Matthews, Jeffrey B

    2005-10-01

    Normal human colonic luminal (NH(4)(+)) concentration ([NH(4)(+)]) ranges from approximately 10 to 100 mM. However, the nature of the effects of NH(4)(+) on transport, as well as NH(4)(+) transport itself, in colonic epithelium is poorly understood. We elucidate here the effects of apical NH(4)(+) on cAMP-stimulated Cl(-) secretion in colonic T84 cells. In HEPES-buffered solutions, 10 mM apical NH(4)(+) had no significant effect on cAMP-stimulated current. In contrast, 10 mM apical NH(4)(+) reduced current within 5 min to 61 +/- 4% in the presence of 25 mM HCO(3)(-). Current inhibition was not simply due to an increase in extracellular K(+)-like cations, in that the current magnitude was 95 +/- 5% with 10 mM apical K(+) and 46 +/- 3% with 10 mM apical NH(4)(+) relative to that with 5 mM apical K(+). We previously demonstrated that inhibition of Cl(-) secretion by basolateral NH(4)(+) occurs in HCO(3)(-)-free conditions and exhibits anomalous mole fraction behavior. In contrast, apical NH(4)(+) inhibition of current in HCO(3)(-) buffer did not show anomalous mole fraction behavior and followed the absolute [NH(4)(+)] in K(+)-NH(4)(+) mixtures, where K(+) concentration + [NH(4)(+)] = 10 mM. The apical NH(4)(+) inhibitory effect was not prevented by 100 microM methazolamide, suggesting no role for apical carbonic anhydrase. However, apical NH(4)(+) inhibition of current was prevented by 10 min of pretreatment of the apical surface with 500 microM DIDS, 100 microM 4,4'-dinitrostilbene-2,2'-disulfonic acid (DNDS), or 25 microM niflumic acid, suggesting a role for NH(4)(+) action through an apical anion exchanger. mRNA and protein for the apical anion exchangers SLC26A3 [downregulated in adenoma (DRA)] and SLC26A6 [putative anion transporter (PAT1)] were detected in T84 cells by RT-PCR and Northern and Western blots. DRA and PAT1 appear to associate with CFTR in the apical membrane. We conclude that the HCO(3)(-) dependence of apical NH(4)(+) inhibition of secretion is

  18. Mycoplasma synoviae has two distinct phase-variable major membrane antigens, one of which is a putative hemagglutinin.

    OpenAIRE

    Noormohammadi, A H; Markham, P. F.; Whithear, K G; Walker, I.D.; Gurevich, V A; Ley, D H; Browning, G F

    1997-01-01

    Mycoplasma synoviae is a major pathogen of poultry, causing synovitis and respiratory infection. A cluster of 45- to 50-kDa membrane proteins is immunodominant in strain WVU-1853. Four distinct proteins were identified in this cluster by high-pressure liquid chromatography. Monoclonal antibodies and monospecific antisera against each established that they fell into two groups, MSPA and MSPB, each containing two members distinguishable by a difference in hydrophobicity. A 25- to 30-kDa membran...

  19. Codon optimisation to improve expression of a Mycobacterium avium ssp. paratuberculosis-specific membrane-associated antigen by Lactobacillus salivarius.

    Science.gov (United States)

    Johnston, Christopher; Douarre, Pierre E; Soulimane, Tewfik; Pletzer, Daniel; Weingart, Helge; MacSharry, John; Coffey, Aidan; Sleator, Roy D; O'Mahony, Jim

    2013-06-01

    Subunit and DNA-based vaccines against Mycobacterium avium ssp. paratuberculosis (MAP) attempt to overcome inherent issues associated with whole-cell formulations. However, these vaccines can be hampered by poor expression of recombinant antigens from a number of disparate hosts. The high G+C content of MAP invariably leads to a codon bias throughout gene expression. To investigate if the codon bias affects recombinant MAP antigen expression, the open reading frame of a MAP-specific antigen MptD (MAP3733c) was codon optimised for expression against a Lactobacillus salivarius host. Of the total 209 codons which constitute MAP3733c, 172 were modified resulting in a reduced G+C content from 61% for the native gene to 32.7% for the modified form. Both genes were placed under the transcriptional control of the PnisA promoter; allowing controlled heterologous expression in L. salivarius. Expression was monitored using fluorescence microscopy and microplate fluorometry via GFP tags translationally fused to the C-termini of the two MptD genes. A > 37-fold increase in expression was observed for the codon-optimised MAP3733synth variant over the native gene. Due to the low cost and improved expression achieved, codon optimisation significantly improves the potential of L. salivarius as an oral vaccine stratagem against Johne's disease. PMID:23620276

  20. Membrane domains and polarized trafficking of sphingolipids

    NARCIS (Netherlands)

    Maier, O; Slimane, TA; Hoekstra, D

    2001-01-01

    The plasma membrane of polarized cells consists of distinct domains, the apical and basolateral membrane that are characterized by a distinct lipid and protein content. Apical protein transport is largely mediated by (glyco)sphingolipid-cholesterol enriched membrane microdomains, so called rafts. In

  1. Apical Localization of Sodium-Dependent Glucose Transporter SGLT1 is Maintained by Cholesterol and Microtubules

    OpenAIRE

    Suzuki, Takeshi; Matsuzaki, Toshiyuki; Hagiwara, Haruo; Aoki, Takeo; Tajika-Takahashi, Yukiko; Takata, Kuniaki

    2006-01-01

    A GFP-labeled sodium-dependent glucose transporter SGLT1 (SGLT-GFP) was transfected into MDCK cells. SGLT-GFP was localized at the apical membrane in confluent cells. When cellular cholesterol was depleted by methyl-β-cyclodextrin (MβCD) treatment, the localization of SGLT-GFP gradually switched from apical to whole plasma membrane. Time-lapse microscopy revealed that the effect of MβCD appeared within 30 min, and that the transition of SGLT-GFP to the whole plasma membrane was completed with...

  2. A transferrin-like GPI-linked iron-binding protein in detergent-insoluble noncaveolar microdomains at the apical surface of fetal intestinal epithelial cells

    DEFF Research Database (Denmark)

    Danielsen, E M; van Deurs, B

    1995-01-01

    ultracryosections of mucosal tissue, the protein was localized to the apical surface of the enterocytes, whereas it was absent from the basolateral plasma membrane. Interestingly, it was mainly found in patches of flat or invaginated apical membrane domains rather than at the surface of microvilli. Caveolae were...

  3. The fast-recycling receptor Megalin defines the apical recycling pathway of epithelial cells.

    Science.gov (United States)

    Perez Bay, Andres E; Schreiner, Ryan; Benedicto, Ignacio; Paz Marzolo, Maria; Banfelder, Jason; Weinstein, Alan M; Rodriguez-Boulan, Enrique J

    2016-01-01

    The basolateral recycling and transcytotic pathways of epithelial cells were previously defined using markers such as transferrin (TfR) and polymeric IgA (pIgR) receptors. In contrast, our knowledge of the apical recycling pathway remains fragmentary. Here we utilize quantitative live-imaging and mathematical modelling to outline the recycling pathway of Megalin (LRP-2), an apical receptor with key developmental and renal functions, in MDCK cells. We show that, like TfR, Megalin is a long-lived and fast-recycling receptor. Megalin enters polarized MDCK cells through segregated apical sorting endosomes and subsequently intersects the TfR and pIgR pathways at a perinuclear Rab11-negative compartment termed common recycling endosomes (CRE). Whereas TfR recycles to the basolateral membrane from CRE, Megalin, like pIgR, traffics to subapical Rab11-positive apical recycling endosomes (ARE) and reaches the apical membrane in a microtubule- and Rab11-dependent manner. Hence, Megalin defines the apical recycling pathway of epithelia, with CRE as its apical sorting station.

  4. Linker Modification Strategies To Control the Prostate-Specific Membrane Antigen (PSMA)-Targeting and Pharmacokinetic Properties of DOTA-Conjugated PSMA Inhibitors.

    Science.gov (United States)

    Benešová, Martina; Bauder-Wüst, Ulrike; Schäfer, Martin; Klika, Karel D; Mier, Walter; Haberkorn, Uwe; Kopka, Klaus; Eder, Matthias

    2016-03-10

    Since prostate-specific membrane antigen (PSMA) is up-regulated in nearly all stages of prostate cancer (PCa), PSMA can be considered as a viable diagnostic biomarker and treatment target in PCa. This project is focused on the development and evaluation of a series of compounds directed against PSMA. The modifications to the linker are designed to improve the binding potential and pharmacokinetics for theranostic application. In addition, the results help to further elucidate the structure-activity relationships (SAR) of the resulting PSMA inhibitors. Both in vitro and in vivo experiments of 18 synthesized PSMA inhibitor variants showed that systematic chemical modification of the linker has a significant impact on the tumor-targeting and pharmacokinetic properties. This approach can lead to an improved management of patients suffering from recurrent prostate cancer by the use of one radiolabeling precursor, which can be radiolabeled either with (68)Ga for diagnosis or with (177)Lu or (225)Ac for therapy. PMID:26878194

  5. Deep-apical tubules: dynamic lipid-raft microdomains in the brush-border region of enterocytes

    DEFF Research Database (Denmark)

    Hansen, Gert H; Pedersen, Jens; Niels-Christiansen, Lise-Lotte;

    2003-01-01

    microdomains. Deep-apical tubules were positioned close to the actin rootlets of adjacent microvilli in the terminal web region, which had a diameter of 50-100 nm, and penetrated up to 1 microm into the cytoplasm. Markers for transcytosis, IgA and the polymeric immunoglobulin receptor, as well as the resident...... lipid raft-containing compartments, but little is otherwise known about these raft microdomains. We therefore studied in closer detail apical lipid-raft compartments in enterocytes by immunogold electron microscopy and biochemical analyses. Novel membrane structures, deep-apical tubules, were visualized...... brush-border enzyme aminopeptidase N, were present in these deep-apical tubules. We propose that deep-apical tubules are a specialized lipid-raft microdomain in the brush-border region functioning as a hub in membrane trafficking at the brush border. In addition, the sensitivity to cholesterol depletion...

  6. A bioinformatics prediction approach towards analyzing the glycosylation, co-expression and interaction patterns of epithelial membrane antigen (EMA/MUC1)

    Energy Technology Data Exchange (ETDEWEB)

    Kalra, Rajkumar S., E-mail: renu-wadhwa@aist.go.jp; Wadhwa, Renu, E-mail: renu-wadhwa@aist.go.jp [Cell Proliferation Research Group and DBT-AIST International Laboratory for Advanced Biomedicine, National Institute of Advanced Industrial Science and Technology (AIST Central 4), 1-1-1 Higashi, Tsukuba, Ibaraki 305-8562 (Japan)

    2015-02-27

    Epithelial membrane antigen (EMA or MUC1) is a heavily glycosylated, type I transmembrane glycoprotein commonly expressed by epithelial cells of duct organs. It has been shown to be aberrantly glycosylated in several diseases including cancer. Protein sequence based annotation and analysis of glycosylation profile of glycoproteins by robust computational and comprehensive algorithms provides possible insights to the mechanism(s) of anomalous glycosylation. In present report, by using a number of bioinformatics applications we studied EMA/MUC1 and explored its trans-membrane structural domain sequence that is widely subjected to glycosylation. Exploration of different extracellular motifs led to prediction of N and O-linked glycosylation target sites. Based on the putative O-linked target sites, glycosylated moieties and pathways were envisaged. Furthermore, Protein network analysis demonstrated physical interaction of EMA with a number of proteins and confirmed its functional involvement in cell growth and proliferation pathways. Gene Ontology analysis suggested an involvement of EMA in a number of functions including signal transduction, protein binding, processing and transport along with glycosylation. Thus, present study explored potential of bioinformatics prediction approach in analyzing glycosylation, co-expression and interaction patterns of EMA/MUC1 glycoprotein.

  7. A bioinformatics prediction approach towards analyzing the glycosylation, co-expression and interaction patterns of epithelial membrane antigen (EMA/MUC1)

    Science.gov (United States)

    Kalra, Rajkumar S.; Wadhwa, Renu

    2015-02-01

    Epithelial membrane antigen (EMA or MUC1) is a heavily glycosylated, type I transmembrane glycoprotein commonly expressed by epithelial cells of duct organs. It has been shown to be aberrantly glycosylated in several diseases including cancer. Protein sequence based annotation and analysis of glycosylation profile of glycoproteins by robust computational and comprehensive algorithms provides possible insights to the mechanism(s) of anomalous glycosylation. In present report, by using a number of bioinformatics applications we studied EMA/MUC1 and explored its trans-membrane structural domain sequence that is widely subjected to glycosylation. Exploration of different extracellular motifs led to prediction of N and O-linked glycosylation target sites. Based on the putative O-linked target sites, glycosylated moieties and pathways were envisaged. Furthermore, Protein network analysis demonstrated physical interaction of EMA with a number of proteins and confirmed its functional involvement in cell growth and proliferation pathways. Gene Ontology analysis suggested an involvement of EMA in a number of functions including signal transduction, protein binding, processing & transport along with glycosylation. Thus, present study explored potential of bioinformatics prediction approach in analyzing glycosylation, co-expression and interaction patterns of EMA/MUC1 glycoprotein.

  8. A bioinformatics prediction approach towards analyzing the glycosylation, co-expression and interaction patterns of epithelial membrane antigen (EMA/MUC1)

    International Nuclear Information System (INIS)

    Epithelial membrane antigen (EMA or MUC1) is a heavily glycosylated, type I transmembrane glycoprotein commonly expressed by epithelial cells of duct organs. It has been shown to be aberrantly glycosylated in several diseases including cancer. Protein sequence based annotation and analysis of glycosylation profile of glycoproteins by robust computational and comprehensive algorithms provides possible insights to the mechanism(s) of anomalous glycosylation. In present report, by using a number of bioinformatics applications we studied EMA/MUC1 and explored its trans-membrane structural domain sequence that is widely subjected to glycosylation. Exploration of different extracellular motifs led to prediction of N and O-linked glycosylation target sites. Based on the putative O-linked target sites, glycosylated moieties and pathways were envisaged. Furthermore, Protein network analysis demonstrated physical interaction of EMA with a number of proteins and confirmed its functional involvement in cell growth and proliferation pathways. Gene Ontology analysis suggested an involvement of EMA in a number of functions including signal transduction, protein binding, processing and transport along with glycosylation. Thus, present study explored potential of bioinformatics prediction approach in analyzing glycosylation, co-expression and interaction patterns of EMA/MUC1 glycoprotein

  9. Apical Ca2+-activated potassium channels in mouse parotid acinar cells.

    Science.gov (United States)

    Almassy, Janos; Won, Jong Hak; Begenisich, Ted B; Yule, David I

    2012-02-01

    Ca(2+) activation of Cl and K channels is a key event underlying stimulated fluid secretion from parotid salivary glands. Cl channels are exclusively present on the apical plasma membrane (PM), whereas the localization of K channels has not been established. Mathematical models have suggested that localization of some K channels to the apical PM is optimum for fluid secretion. A combination of whole cell electrophysiology and temporally resolved digital imaging with local manipulation of intracellular [Ca(2+)] was used to investigate if Ca(2+)-activated K channels are present in the apical PM of parotid acinar cells. Initial experiments established Ca(2+)-buffering conditions that produced brief, localized increases in [Ca(2+)] after focal laser photolysis of caged Ca(2+). Conditions were used to isolate K(+) and Cl(-) conductances. Photolysis at the apical PM resulted in a robust increase in K(+) and Cl(-) currents. A localized reduction in [Ca(2+)] at the apical PM after photolysis of Diazo-2, a caged Ca(2+) chelator, resulted in a decrease in both K(+) and Cl(-) currents. The K(+) currents evoked by apical photolysis were partially blocked by both paxilline and TRAM-34, specific blockers of large-conductance "maxi-K" (BK) and intermediate K (IK), respectively, and almost abolished by incubation with both antagonists. Apical TRAM-34-sensitive K(+) currents were also observed in BK-null parotid acini. In contrast, when the [Ca(2+)] was increased at the basal or lateral PM, no increase in either K(+) or Cl(-) currents was evoked. These data provide strong evidence that K and Cl channels are similarly distributed in the apical PM. Furthermore, both IK and BK channels are present in this domain, and the density of these channels appears higher in the apical versus basolateral PM. Collectively, this study provides support for a model in which fluid secretion is optimized after expression of K channels specifically in the apical PM.

  10. MS-H: a novel proteomic approach to isolate and type the E. coli H antigen using membrane filtration and liquid chromatography-tandem mass spectrometry (LC-MS/MS.

    Directory of Open Access Journals (Sweden)

    Keding Cheng

    Full Text Available Serotyping is the long-standing gold standard method to determine E. coli H antigens; however, this method requires a panel of H-antigen specific antibodies and often culture-based induction of the H-antigen flagellar motility. In this study, a rapid and accurate method to isolate and identify the Escherichia coli (E. coli H flagellar antigen was developed using membrane filtration and liquid chromatography-tandem mass spectrometry (LC-MS/MS. Flagella were isolated from pure culture, digested with trypsin, and then subjected to LC-MS/MS using one of two systems (Agilent-nano-LC-QSTAR XL or Proxeon-nano-LC-LTQ-Orbitrap XL. The resulting peptide sequence data were searched against a custom E. coli flagella/H antigen database. This approach was evaluated using flagella isolated from reference E. coli strains representing all 53 known H antigen types and 41 clinical E. coli strains. The resulting LC-MS/MS classifications of H antigen types (MS-H were concordant with the known H serogroup for all 53 reference types, and of 41 clinical isolates tested, 38 (92.7% were concordant with the known H serogroup. MS-H clearly also identified two clinical isolates (4.9% that were untypeable by serotyping. Notably, successful detection and classification of flagellar antigens with MS-H did not generally require induction of motility, establishing this proteomic approach as more rapid and cost-effective than traditional methods, while providing equitable specificity for typing E. coli H antigens.

  11. Regulated delayed expression of rfaH in an attenuated Salmonella enterica serovar typhimurium vaccine enhances immunogenicity of outer membrane proteins and a heterologous antigen.

    Science.gov (United States)

    Kong, Qingke; Liu, Qing; Roland, Kenneth L; Curtiss, Roy

    2009-12-01

    RfaH is a transcriptional antiterminator that reduces the polarity of long operons encoding secreted and surface-associated cell components of Salmonella enterica serovar Typhimurium, including O antigen and lipopolysaccharide core sugars. A DeltarfaH mutant strain is attenuated in mice (50% lethal dose [LD(50)], >10(8) CFU). To examine the potential for using rfaH in conjunction with other attenuating mutations, we designed a series of strains in which we replaced the native rfaH promoter with the tightly regulated arabinose-dependent araC P(BAD) promoter so that rfaH expression was dependent on exogenously supplied arabinose provided during in vitro growth. Following colonization of host lymphoid tissues, where arabinose was not available, the P(BAD) promoter was no longer active and rfaH was not expressed. In the absence of RfaH, O antigen and core sugars were not synthesized. We constructed three mutant strains that expressed different levels of RfaH by altering the ribosome-binding sequence and start codon. One mutation, DeltaP(rfaH178), was introduced into the attenuated vaccine strain chi9241 (DeltapabA DeltapabB DeltaasdA) expressing the pneumococcal surface protein PspA from an Asd(+) balanced-lethal plasmid. Mice immunized with this strain and boosted 4 weeks later induced higher levels of serum immunoglobulin G specific for PspA and for outer membrane proteins from other enteric bacteria than either an isogenic DeltarfaH derivative or the isogenic RfaH(+) parent. Eight weeks after primary oral immunization, mice were challenged with 200 LD(50) of virulent Streptococcus pneumoniae WU2. Immunization with DeltaP(rfaH178) mutant strains led to increased levels of protection compared to that of the parent chi9241 and of a DeltarfaH derivative of chi9241.

  12. [(18)F]Fluorobenzoyllysinepentanedioic Acid Carbamates: New Scaffolds for Positron Emission Tomography (PET) Imaging of Prostate-Specific Membrane Antigen (PSMA).

    Science.gov (United States)

    Yang, Xing; Mease, Ronnie C; Pullambhatla, Mrudula; Lisok, Ala; Chen, Ying; Foss, Catherine A; Wang, Yuchuan; Shallal, Hassan; Edelman, Hannah; Hoye, Adam T; Attardo, Giorgio; Nimmagadda, Sridhar; Pomper, Martin G

    2016-01-14

    Radiolabeled urea-based low-molecular weight inhibitors of the prostate-specific membrane antigen (PSMA) are under intense investigation as imaging and therapeutic agents for prostate and other cancers. In an effort to provide agents with less nontarget organ uptake than the ureas, we synthesized four (18)F-labeled inhibitors of PSMA based on carbamate scaffolds. 4-Bromo-2-[(18)F]fluorobenzoyllysineoxypentanedioic acid (OPA) carbamate [(18)F]23 and 4-iodo-2-[(18)F]fluorobenzoyllysine OPA carbamate [(18)F]24 in particular exhibited high target-selective uptake in PSMA+ PC3 PIP tumor xenografts, with tumor-to-kidney ratios of >1 by 4 h postinjection, an important benchmark. Because of its high tumor uptake (90% injected dose per gram of tissue at 2 h postinjection) and high tumor-to-organ ratios, [(18)F]23 is promising for clinical translation. Prolonged tumor-specific uptake demonstrated by [(18)F]24, which did not reach equilibrium during the 4 h study period, suggests carbamates as alternative scaffolds for mitigating dose to nontarget tissues. PMID:26629713

  13. Partial Characterisation of Salmonella gallinarum Clinical Isolate and Expression of Its Antigenic Outer Membrane Protein C (OmpC Gene In Planta

    Directory of Open Access Journals (Sweden)

    Ee Leen Pang

    2015-05-01

    Full Text Available Fowl typhoid’s epidemiology and disease intervention have been extensively studied since 1950's owing to its high mortality and morbidity rates. Even up-to-date, outbreaks are incessantly haunting poultry industries of major continents. Salmonella gallinarum, the etiologic agent of fowl typhoid, was used to develop a series of vaccination regime. However, treatments are gradually losing effectiveness due to residual virulence from mutated strains and rapid evolution of multi-drug resistance isolates. Hence, in planta subunit vaccine production is proposed to surpass current limitations. The homotrimeric osmoporin (outer membrane protein C is a potent candidate antigen that confers momentous stimulation of humoral and cell-mediated immune responses in broilers. This research signified the potential development of a plant-expressed OmpC immunogen. The project scope embarked on the identification of S. gallinarum clinical isolate, construction of expression cassette and delivery of constructs into Nicotiana benthamiana via agroinfiltration. The OmpC transcripts and proteins were detected successfully at the molecular weights of ~1.002 kbp and ~35 kDa, respectively. These preliminary findings pave the feasibility of biomanufacturing a safe and cost-effective fowl typhoid vaccine that would confer multi-protection against other significant Salmonella infections attributed to the high sequence homology of the OmpC gene. Speed improvement is demonstrated and transient expression appears to outperform conventional platforms in expediting vaccine production for an emerging pandemic strain.

  14. Surgical treatments for vaginal apical prolapse.

    Science.gov (United States)

    Kong, Mi Kyung; Bai, Sang Wook

    2016-07-01

    Pelvic organ prolapse is a common condition, occurring in up to 11% of women in the United States. Often, pelvic organ prolapse recurs after surgery; when it recurs after hysterectomy, it frequently presents as vaginal apical prolapse. There are many different surgical treatments for vaginal apical prolapse; among them, abdominal sacral colpopexy is considered the gold standard. However, recent data reveal that other surgical procedures also result in good outcome. This review discusses the various surgical treatments for vaginal apical prolapse including their risks and benefits. PMID:27462591

  15. Apical Na+ permeability of frog skin during serosal Cl- replacement.

    Science.gov (United States)

    Leibowich, S; DeLong, J; Civan, M M

    1988-05-01

    Gluconate substitution for serosal Cl- reduces the transepithelial short-circuit current (Isc) and depolarizes short-circuited frog skins. These effects could result either from inhibition of basolateral K+ conductance, or from two actions to inhibit both apical Na+ permeability (PapNa) and basolateral pump activity. We have addressed this question by studying whole-and split-thickness frog skins. Intracellular Na+ concentration (CcNa) and PapNa have been monitored by measuring the current-voltage relationship for apical Na+ entry. This analysis was conducted by applying trains of voltage pulses, with pulse durations of 16 to 32 msec. Estimates of PapNa and CcNa were not detectably dependent on pulse duration over the range 16 to 80 msec. Serosal Cl- replacement uniformly depolarized short-circuited tissues. The depolarization was associated with inhibition of Isc across each split skin, but only occasionally across the whole-thickness preparations. This difference may reflect the better ionic exchange between the bulk medium and the extracellular fluid in contact with the basolateral membranes, following removal of the underlying dermis in the split-skin preparations. PapNa was either unchanged or increased, and CcNa either unchanged or reduced after the anionic replacement. These data are incompatible with the concept that serosal Cl- replacement inhibits PapNa and Na,K-pump activity. Gluconate substitution likely reduces cell volume, triggering inhibition of the basolateral K+ channels, consistent with the data and conclusions of S.A. Lewis, A.G. Butt, M.J. Bowler, J.P. Leader and A.D.C. Macknight (J. Membrane Biol. 83:119-137, 1985) for toad bladder. The resulting depolarization reduces the electrical force favoring apical Na+ entry. The volume-conductance coupling serves to conserve volume by reducing K+ solute loss. Its molecular basis remains to be identified. PMID:2458472

  16. Apical Na+ permeability of frog skin during serosal Cl- replacement.

    Science.gov (United States)

    Leibowich, S; DeLong, J; Civan, M M

    1988-05-01

    Gluconate substitution for serosal Cl- reduces the transepithelial short-circuit current (Isc) and depolarizes short-circuited frog skins. These effects could result either from inhibition of basolateral K+ conductance, or from two actions to inhibit both apical Na+ permeability (PapNa) and basolateral pump activity. We have addressed this question by studying whole-and split-thickness frog skins. Intracellular Na+ concentration (CcNa) and PapNa have been monitored by measuring the current-voltage relationship for apical Na+ entry. This analysis was conducted by applying trains of voltage pulses, with pulse durations of 16 to 32 msec. Estimates of PapNa and CcNa were not detectably dependent on pulse duration over the range 16 to 80 msec. Serosal Cl- replacement uniformly depolarized short-circuited tissues. The depolarization was associated with inhibition of Isc across each split skin, but only occasionally across the whole-thickness preparations. This difference may reflect the better ionic exchange between the bulk medium and the extracellular fluid in contact with the basolateral membranes, following removal of the underlying dermis in the split-skin preparations. PapNa was either unchanged or increased, and CcNa either unchanged or reduced after the anionic replacement. These data are incompatible with the concept that serosal Cl- replacement inhibits PapNa and Na,K-pump activity. Gluconate substitution likely reduces cell volume, triggering inhibition of the basolateral K+ channels, consistent with the data and conclusions of S.A. Lewis, A.G. Butt, M.J. Bowler, J.P. Leader and A.D.C. Macknight (J. Membrane Biol. 83:119-137, 1985) for toad bladder. The resulting depolarization reduces the electrical force favoring apical Na+ entry. The volume-conductance coupling serves to conserve volume by reducing K+ solute loss. Its molecular basis remains to be identified.

  17. Use of radioimmune assay in investigating reagents to be used in the immunocytochemical localization of hepatitis B surface antigen in immune complexes in the kidney of patients with membranous nephropathy and Australia antigenaemia

    Energy Technology Data Exchange (ETDEWEB)

    Wolfe-Coote, S. (South African Medical Research Council, Tygerberg (South Africa). Inst. for Electron Microscopy)

    1983-09-01

    Radioimmune assay (RIA) was used to investigate the effect of fixatives on antigenicity of the hepatitis B surface antigen (HBsAg) and the effect of pronase on the elution of antibody (Ab) from the HBsAg-Ab complex. The effect of pronase on Ab elution was also tested on sections of kidney from a patient with the immune complex disease systemic lupus erythematosus (SLE). Immunoglobulin G (IgG) was located in pronase treated and untreated sections using the indirect immunoperoxidase technique. Glutareldehyde was shown to be the fixative of choice for studies involving HBsAg. All fixatives were shown to have less effect on antigenicity at 4/sup 0/C than at room temperature. Osmium tetroxide reduced antigenicity to one-third, even at 4/sup 0/C. RIA and SLE kidney section studies showed that Ab was eluted from immune complexes by pronase. Pre-fixation of the antigen (Ag) by glutaraldehyde appears to have no effect on the final elution, although fixation after pronase treatment seemed to enhance the elution effects. The availability of an RIA kit with HBsAg- and Ab-coated beads was of great assistance in evaluating reagents to be used in immunoperoxidase studies of HBsAg in immune complexes of patients with membranous nephropathy and Australia antigenaemia.

  18. Cochlear outer hair cells undergo an apical circumference remodeling constrained by the hair bundle shape.

    Science.gov (United States)

    Etournay, Raphaël; Lepelletier, Léa; Boutet de Monvel, Jacques; Michel, Vincent; Cayet, Nadège; Leibovici, Michel; Weil, Dominique; Foucher, Isabelle; Hardelin, Jean-Pierre; Petit, Christine

    2010-04-01

    Epithelial cells acquire diverse shapes relating to their different functions. This is particularly relevant for the cochlear outer hair cells (OHCs), whose apical and basolateral shapes accommodate the functioning of these cells as mechano-electrical and electromechanical transducers, respectively. We uncovered a circumferential shape transition of the apical junctional complex (AJC) of OHCs, which occurs during the early postnatal period in the mouse, prior to hearing onset. Geometric analysis of the OHC apical circumference using immunostaining of the AJC protein ZO1 and Fourier-interpolated contour detection characterizes this transition as a switch from a rounded-hexagon to a non-convex circumference delineating two lateral lobes at the neural side of the cell, with a negative curvature in between. This shape tightly correlates with the 'V'-configuration of the OHC hair bundle, the apical mechanosensitive organelle that converts sound-evoked vibrations into variations in cell membrane potential. The OHC apical circumference remodeling failed or was incomplete in all the mouse mutants affected in hair bundle morphogenesis that we tested. During the normal shape transition, myosin VIIa and myosin II (A and B isoforms) displayed polarized redistributions into and out of the developing lobes, respectively, while Shroom2 and F-actin transiently accumulated in the lobes. Defects in these redistributions were observed in the mutants, paralleling their apical circumference abnormalities. Our results point to a pivotal role for actomyosin cytoskeleton tensions in the reshaping of the OHC apical circumference. We propose that this remodeling contributes to optimize the mechanical coupling between the basal and apical poles of mature OHCs.

  19. A novel recombinant bivalent outer membrane protein of Vibrio vulnificus and Aeromonas hydrophila as a vaccine antigen of American eel (Anguilla rostrata).

    Science.gov (United States)

    SongLin, Guo; PanPan, Lu; JianJun, Feng; JinPing, Zhao; Peng, Lin; LiHua, Duan

    2015-04-01

    The immogenicity of a novel vaccine antigen was evaluated after immunized American eels (Anguilla rostrata) with a recombinant bivalent expressed outer membrane protein (OMP) of Vibrio vulnificus and Aeromonas hydrophila. Three groups of eels were intraperitoneal (i.p) injected with phosphate-buffered saline (PBS group), formaline-killed-whole-cell (FKC) of A. hydrophila and V. vulnificus (FKC group) or the bivalent OMP (OMP group). On 14, 21, 28 and 42 days post-vaccination respectively, proliferation of the whole blood cells, titers of specific antibody and lysozyme activities of experimental eels were detected. On 28 day post-vaccination, eels from three groups were challenged by i.p injection of live A. hydrophila or V. vulnificus. The results showed that, compared with the PBS group, proliferation of whole blood cells in OMP group was significant enhanced on 28 days, and the serum titers of anti-A.hydrophila and anti-V. vulnificus antibody in eels of FKC and OMP group were significant increased on 14, 21 and 28d. Lysozyme Activities in serum, skin mucus, liver and kidney were significant changed between the three groups. Relative Percent Survival (RPS) after challenged A. hydrophila in KFC vs. PBS group and OMP vs. PBS group were 62.5% and 50% respectively, and the RPS challenged V. vulnificus in FKC and OMP vs. PBS group were 37.5% and 50% respectively. These results suggest that American eels immunized with the bivalent OMP would positively affect specific as well as non-specific immune parameters and protect against infection by the two pathogens in fresh water farming. PMID:25655329

  20. Radiolabeling of anti-human prostatic specific membrane antigen antibody with 99Tcm and its biodistribution in nude mice bearing human prostate cancer

    International Nuclear Information System (INIS)

    Objective: To study the binding affinity of 99Tcm labeled anti-human prostatic specific membrane antigen (PSMA) monoclonal antibody (McAb) J591 to prostate cancer cells and the biodistribution of 99Tcm-J591 in nude mice bearing human prostate cancer. Methods: The McAb J591 was labeled with vTcm by improved Schwarz method and the labeled McAb was purified by Sephadex G-50. The binding affinity of J591 with prostate cancer cells was measured by Flow Cytometry. The nude mice bearing PSMA-positive C4-2 prostate carcinoma xenografts were served as experiment groups, mice with PSMA-negative pc3 tumors served as controls. The biodistribution of 99Tcm-J591 were carried out in both model nude mice. Results: The radiolabeling efficiency of 99Tcm-J591 was 78.9±6.2%, and radiochemical purity was more than 90% after purification. The 99Tcm-J591 showed a good combination with PSMA-positive C4-2 cells and no combination with PSMA-negative PC3 cells in vitro. The biodistribution results showed that 99Tcm-J591 was accumulated in tumor tissue during the 2-24 hours after injection in experiment groups, and no significant uptake in control group. The uptake of 99Tcm-J591 in tumor tissue reached a maximum 15.91±5.16 % ID/g in experimental group at 12h post-injection. There was a significant difference compared with controls (P0.05). Conclusion: The monoclonal antibody J591 exhibits an excellent immuno-reactivity and tumor targeting property, and it may be used in diagnosis and target therapy of prostate cancer. (authors)

  1. Activation of an apical Cl- conductance by Ca2+ ionophores in cystic fibrosis airway epithelia.

    Science.gov (United States)

    Willumsen, N J; Boucher, R C

    1989-02-01

    Cystic fibrosis (CF) airway epithelia express a defect in adenosine 3',5'-cyclic monophosphate (cAMP)-dependent regulation of apical membrane Cl- channels. Recent patch-clamp studies have raised the possibility that Ca2+ -dependent mechanisms for the activation of Cl- secretion may be preserved in CF airway epithelia. To determine 1) whether intact normal (N1) and CF airway epithelia exhibit a Ca2+ -dependent mechanism for activation of Cl- secretion and 2) whether Ca2+ -dependent mechanism for activation of Cl- secretion and 2) whether Ca2+ -dependent mechanisms initiate Cl- secretion via activation of an apical membrane Cl- conductance (GCl-), nasal epithelia from N1 and CF subjects were cultured on collagen membranes, and responses to isoproterenol or Ca2- ionophores [A23187 10(-6) M; ionomycin (10(-5)M)] were measured with transepithelial and intracellular techniques. Isoproterenol induced activation of an apical membrane GCl- in N1 cultures but was ineffective in CF. In contrast, in both N1 and CF amiloride-pretreated cultures, A23187 induced an increase in the equivalent short-circuit current that was associated with an activation of an apical membrane Gc1- and was bumetanide inhibitable. A23187 addition during superfusion of the lumen with a low Cl- (3 mM) solution reduced intracellular Cl- activity of CF cells. A Ca2+ ionophore of different selectivity properties, ionomycin, was also an effective Cl- secretagogue in both N1 and CF cultures. We conclude that 1) the A23187 induced Cl- secretion via activation of an apical GCl- in N1 human nasal epithelium, and 2) in contrast to an isoproterenol-dependent path, a Ca2+ -dependent path for GCl- activation is preserved in CF epithelia. PMID:2465689

  2. A Comparative Study of Apical Healing of Open Apices Using MTA and Ca(OH2 Apical Plugs in Cats

    Directory of Open Access Journals (Sweden)

    M. H. Zarrabi

    2005-06-01

    Full Text Available Statement of problem: Endodontic treatment of necrotic teeth with open apices is a challenge. After ruling out surgery as a treatment scheme and introduction of the multivisit apexification which in turn had its disadvantages, apical plug seems to be a suitable substitute treatment plan for such cases. Apical plug makes the treatment through formation of a barrier against the obturating material in a single visit.Purpose: The purpose of this study was to compare histologically the periapical healing using MTA and calcium hydroxide apical plugs after intervals of 4 and 12 weeks in cats.Materials and Methods: In this clinical trial study 64 canines of 16 healthy and mature cats were divided into 3 groups after a periapical lesion formation by over instrumentation in the apical area with files up to no.120. The first group included 24 teeth on which MTA apical plug was applied. The second group included 24 teeth on which Ca (OH 2 apical plug was applied. In both groups the canals were filled with gutta percha and sealer. The third group included 16 control teeth whose canals were left empty after instrumentation and debridement. The access cavities of all teeth were sealed with varnish and amalgam and the vital perfusion of cats was performed in 4 and 12 week intervals. Statistical analysis was established by χ2 and independence test.Results: After 4 weeks, periapical healing in the first group was 90%, in the second group 80% and in the third group, it was only 12.5 %. After 12 weeks, periapical healing occurred in 100% of the MTA group, while it was 57.1% in the second and 40%in the third group .Generally, in the study of histological parameters of healing, no statistical significant difference was observed between the 2 experimental groups,although the MTA group results were much better than the Ca (OH 2 group especially at 12 weeks.Conclusion: The use of MTA apical plug is more effective than Ca (OH 2 in treatment of necrotic teeth with open

  3. Genetic diversity and population structure of genes encoding vaccine candidate antigens of Plasmodium vivax

    Directory of Open Access Journals (Sweden)

    Chenet Stella M

    2012-03-01

    Full Text Available Abstract Background A major concern in malaria vaccine development is genetic polymorphisms typically observed among Plasmodium isolates in different geographical areas across the world. Highly polymorphic regions have been observed in Plasmodium falciparum and Plasmodium vivax antigenic surface proteins such as Circumsporozoite protein (CSP, Duffy-binding protein (DBP, Merozoite surface protein-1 (MSP-1, Apical membrane antigen-1 (AMA-1 and Thrombospondin related anonymous protein (TRAP. Methods Genetic variability was assessed in important polymorphic regions of various vaccine candidate antigens in P. vivax among 106 isolates from the Amazon Region of Loreto, Peru. In addition, genetic diversity determined in Peruvian isolates was compared to population studies from various geographical locations worldwide. Results The structured diversity found in P. vivax populations did not show a geographic pattern and haplotypes from all gene candidates were distributed worldwide. In addition, evidence of balancing selection was found in polymorphic regions of the trap, dbp and ama-1 genes. Conclusions It is important to have a good representation of the haplotypes circulating worldwide when implementing a vaccine, regardless of the geographic region of deployment since selective pressure plays an important role in structuring antigen diversity.

  4. Apical root resorption in orthodontically treated adults.

    Science.gov (United States)

    Baumrind, S; Korn, E L; Boyd, R L

    1996-09-01

    This study analyzed the relationship in orthodontically treated adults between upper central incisor displacement measured on lateral cephalograms and apical root resorption measured on anterior periapical x-ray films. A multiple linear regression examined incisor displacements in four directions (retraction, advancement, intrusion, and extrusion) as independent variables, attempting to account for observed differences in the dependent variable, resorption. Mean apical resorption was 1.36 mm (sd +/- 1.46, n = 73). Mean horizontal displacement of the apex was -0.83 mm (sd +/- 1.74, n = 67); mean vertical displacement was 0.19 mm (sd +/- 1.48, n = 67). The regression coefficients for the intercept and for retraction were highly significant; those for extrusion, intrusion, and advancement were not. At the 95% confidence level, an average of 0.99 mm (se = +/- 0.34) of resorption was implied in the absence of root displacement and an average of 0.49 mm (se = +/- 0.14) of resorption was implied per millimeter of retraction. R2 for all four directional displacement variables (DDVs) taken together was only 0.20, which implied that only a relatively small portion of the observed apical resorption could be accounted for by tooth displacement alone. In a secondary set of univariate analyses, the associations between apical resorption and each of 14 additional treatment-related variables were examined. Only Gender, Elapsed Time, and Total Apical Displacement displayed statistically significant associations with apical resorption. Additional multiple regressions were then performed in which the data for each of these three statistically significant variables were considered separately, with the data for the four directional displacement variables. The addition of information on Elapsed Time or Total Apical Displacement did not explain a significant additional portion of the variability in apical resorption. On the other hand, the addition of information on Gender to the

  5. Apical domain polarization localizes actin-myosin activity to drive ratchet-like apical constriction.

    Science.gov (United States)

    Mason, Frank M; Tworoger, Michael; Martin, Adam C

    2013-08-01

    Apical constriction promotes epithelia folding, which changes tissue architecture. During Drosophila gastrulation, mesoderm cells exhibit repeated contractile pulses that are stabilized such that cells apically constrict like a ratchet. The transcription factor Twist is required to stabilize cell shape. However, it is unknown how Twist spatially coordinates downstream signals to prevent cell relaxation. We find that during constriction, Rho-associated kinase (Rok) is polarized to the middle of the apical domain (medioapical cortex), separate from adherens junctions. Rok recruits or stabilizes medioapical myosin II (Myo-II), which contracts dynamic medioapical actin cables. The formin Diaphanous mediates apical actin assembly to suppress medioapical E-cadherin localization and form stable connections between the medioapical contractile network and adherens junctions. Twist is not required for apical Rok recruitment, but instead polarizes Rok medioapically. Therefore, Twist establishes radial cell polarity of Rok/Myo-II and E-cadherin and promotes medioapical actin assembly in mesoderm cells to stabilize cell shape fluctuations.

  6. Serum Concentrations of Antibodies against Outer Membrane Protein P6, Protein D, and T- and B-Cell Combined Antigenic Epitopes of Nontypeable Haemophilus influenzae in Children and Adults of Different Ages.

    Science.gov (United States)

    Hua, Chun-Zhen; Hu, Wei-Lin; Shang, Shi-Qiang; Li, Jian-Ping; Hong, Li-Quan; Yan, Jie

    2016-02-01

    Nontypeable Haemophilus influenzae (NTHi) is one of the most common etiologies of acute otitis media, rhinosinusitis, and pneumonia. Outer membrane proteins (OMPs) are the main focus in new vaccine development against NTHi, as the H. influenzae type b (Hib) vaccine does not cover noncapsulated NTHi. The OMPs P6 and protein D are the most promising candidate antigens for an NTHi vaccine, and low antibody levels against them in serum may be correlated with infection caused by NTHi. In the current study, we measured the antibody titers against P6, protein D, and their T- and B-cell combined peptide epitopes in healthy individuals of different ages. We found that children B-cell combined antigenic epitopes. Antibody titers increased at ages 1 to 6 months, peaked at 7 months to 3 years, and remained high at 4 to 6 years. The antibody titers started to decrease after 6 years and were the lowest in the 21- to 30-year group. The geometric mean titers (GMTs) of T- and B-cell combined antigenic epitopes in P6 and protein D were positively correlated with those of the protein antigens. Among 12 peptides tested, P6-61, P6-123, and protein D-167 epitopes were better recognized than others in human serum. These findings might contribute to the development of an effective serotype-independent vaccine for H. influenzae. PMID:26677200

  7. Analyses of Interactions Between Heparin and the Apical Surface Proteins of Plasmodium falciparum

    Science.gov (United States)

    Kobayashi, Kyousuke; Takano, Ryo; Takemae, Hitoshi; Sugi, Tatsuki; Ishiwa, Akiko; Gong, Haiyan; Recuenco, Frances C.; Iwanaga, Tatsuya; Horimoto, Taisuke; Akashi, Hiroomi; Kato, Kentaro

    2013-11-01

    Heparin, a sulfated glycoconjugate, reportedly inhibits the blood-stage growth of the malaria parasite Plasmodium falciparum. Elucidation of the inhibitory mechanism is valuable for developing novel invasion-blocking treatments based on heparin. Merozoite surface protein 1 has been reported as a candidate target of heparin; however, to better understand the molecular mechanisms involved, we characterized the molecules that bind to heparin during merozoite invasion. Here, we show that heparin binds only at the apical tip of the merozoite surface and that multiple heparin-binding proteins localize preferentially in the apical organelles. To identify heparin-binding proteins, parasite proteins were fractionated by means of heparin affinity chromatography and subjected to immunoblot analysis with ligand-specific antibodies. All tested members of the Duffy and reticulocyte binding-like families bound to heparin with diverse affinities. These findings suggest that heparin masks the apical surface of merozoites and blocks interaction with the erythrocyte membrane after initial attachment.

  8. Preclinical evaluation of BAY 1075553, a novel {sup 18}F-labelled inhibitor of prostate-specific membrane antigen for PET imaging of prostate cancer

    Energy Technology Data Exchange (ETDEWEB)

    Lesche, Ralf; Kettschau, Georg; Gromov, Alexey V.; Boehnke, Niels; Borkowski, Sandra; Moenning, Ursula; Doehr, Olaf; Graham, Keith [Global Drug Discovery, Bayer Healthcare, Berlin, Germany, Berlin (Germany); Hegele-Hartung, Christa [Global Drug Discovery, Bayer Healthcare, Wuppertal, Germany, Wuppertal (Germany); Dinkelborg, Ludger M. [Global Drug Discovery, Bayer Healthcare, Berlin, Germany, Berlin (Germany); Piramal Imaging GmbH, Berlin (Germany)

    2014-01-15

    Prostate-specific membrane antigen (PSMA) is a transmembrane protein overexpressed in prostate cancer and is therefore being explored as a biomarker for diagnosing and staging of the disease. Here we report preclinical data on BAY 1075553 (a 9:1 mixture of (2S,4S)- and (2R,4S)-2-[{sup 18}F]fluoro-4-phosphonomethyl-pentanedioic acid), a novel {sup 18}F-labelled small molecule inhibitor of PSMA enzymatic activity, which can be efficiently synthesized from a direct radiolabelling precursor. The {sup 18}F-radiolabelled stereoisomers of 2-[{sup 18}F]fluoro-4-(phosphonomethyl)-pentanedioic acid were synthesized from their respective isomerically pure precursors dimethyl 2-{[bis(benzyloxy)phosphoryl ]methyl}-4-(tosyloxy)pentanedioate. In vivo positron emission tomography (PET) imaging and biodistribution studies were conducted in mice bearing LNCaP, 22Rv1 and PC-3 tumours. Pharmacokinetic parameters and dosimetry estimates were calculated based on biodistribution studies in rodents. For non-clinical safety assessment (safety pharmacology, toxicology) to support a single-dose human microdose study, off-target effects in vitro, effects on vital organ functions (cardiovascular in dogs, nervous system in rats), mutagenicity screens and an extended single-dose study in rats were conducted with the non-radioactive racemic analogue of BAY 1075553. BAY 1075553 showed high tumour accumulation specific to PSMA-positive tumour-bearing mice and was superior to other stereoisomers tested. Fast clearance of BAY 1075553 resulted overall in low background signals in other organs except for high uptake into kidney and bladder which was mainly caused by renal elimination of BAY 1075553. A modest uptake into bone was observed which decreased over time indicating organ-specific uptake as opposed to defluorination of BAY 1075553 in vivo. Biodistribution studies found highest organ doses for kidneys and the urinary bladder wall resulting in a projected effective dose (ED) in humans of 0.0219 m

  9. A geographical study of antibodies to membrane antigens of HSV-2-infected cells and HSV-2-specific antibodies in patients with cervical cancer.

    Science.gov (United States)

    Mendis, L N; Best, J M; Senarath, L; Chiphangwi, J; Vestergaard, B F; Banatvala, J E

    1981-11-15

    Sera was obtained from patients with squamous carcinoma of the cervix from Great Britain (29), Sri Lanka (32), Malawi (27), and Sudan (27), and from controls from all countries except Sudan. Controls were matched for ethnic origin, age and social class. Sera were tested by indirect immunofluorescence (IF) for IgG and IgA antibodies to membrane antigens (MA) and IgA antibodies to VCA of HSV-2 infected cells. Compared with controls, IgA antibodies to MA (IgA anti-MA) were detected more frequently and at higher titres in all groups of patients. However, there was no significant difference in prevalence of these antibodies at titres greater than or equal to 1:4 between Malawian patients and controls, although a significantly higher proportion of patients had IgA anti-MA titres of greater than or equal to 1:16. In contrast, IgA anti-VCA did not distinguish patients from controls. More than 90% of both patients and controls from all countries had IgG antibodies to MA (IgA anti-MA). Malawian patients had a significantly higher geometric mean titre (GMT) of IgG anti-MA than controls and both patients and controls had significantly higher GMTs than their counterparts from other countries. The variation between herpes IgG anti-MA titres in subjects from different countries did not reflect differences in serum immunoglobulin levels and similar variations in titre were not seen in rubella and measles HAI titres. Among the patients there was a geographical variation in the prevalence of HSV-2 specific antibodies detected by ELISA, which varied from 52% among British and Sudanese patients to 73% among Malawian patients. Even when adjustment was made for possible false negative results, there were between 10 and 31% patients without HSV-2-specific antibodies, although only 2 of 103 (1.9%) patients had neither HSV-1 nor HSV-2 antibodies. The association of HSV-2 with cervical carcinoma appeared to vary with age.

  10. Preclinical evaluation of BAY 1075553, a novel 18F-labelled inhibitor of prostate-specific membrane antigen for PET imaging of prostate cancer

    International Nuclear Information System (INIS)

    Prostate-specific membrane antigen (PSMA) is a transmembrane protein overexpressed in prostate cancer and is therefore being explored as a biomarker for diagnosing and staging of the disease. Here we report preclinical data on BAY 1075553 (a 9:1 mixture of (2S,4S)- and (2R,4S)-2-[18F]fluoro-4-phosphonomethyl-pentanedioic acid), a novel 18F-labelled small molecule inhibitor of PSMA enzymatic activity, which can be efficiently synthesized from a direct radiolabelling precursor. The 18F-radiolabelled stereoisomers of 2-[18F]fluoro-4-(phosphonomethyl)-pentanedioic acid were synthesized from their respective isomerically pure precursors dimethyl 2-{[bis(benzyloxy)phosphoryl ]methyl}-4-(tosyloxy)pentanedioate. In vivo positron emission tomography (PET) imaging and biodistribution studies were conducted in mice bearing LNCaP, 22Rv1 and PC-3 tumours. Pharmacokinetic parameters and dosimetry estimates were calculated based on biodistribution studies in rodents. For non-clinical safety assessment (safety pharmacology, toxicology) to support a single-dose human microdose study, off-target effects in vitro, effects on vital organ functions (cardiovascular in dogs, nervous system in rats), mutagenicity screens and an extended single-dose study in rats were conducted with the non-radioactive racemic analogue of BAY 1075553. BAY 1075553 showed high tumour accumulation specific to PSMA-positive tumour-bearing mice and was superior to other stereoisomers tested. Fast clearance of BAY 1075553 resulted overall in low background signals in other organs except for high uptake into kidney and bladder which was mainly caused by renal elimination of BAY 1075553. A modest uptake into bone was observed which decreased over time indicating organ-specific uptake as opposed to defluorination of BAY 1075553 in vivo. Biodistribution studies found highest organ doses for kidneys and the urinary bladder wall resulting in a projected effective dose (ED) in humans of 0.0219 mSv/MBq. Non

  11. CAMSAP3 orients the apical-to-basal polarity of microtubule arrays in epithelial cells.

    Science.gov (United States)

    Toya, Mika; Kobayashi, Saeko; Kawasaki, Miwa; Shioi, Go; Kaneko, Mari; Ishiuchi, Takashi; Misaki, Kazuyo; Meng, Wenxiang; Takeichi, Masatoshi

    2016-01-12

    Polarized epithelial cells exhibit a characteristic array of microtubules that are oriented along the apicobasal axis of the cells. The minus-ends of these microtubules face apically, and the plus-ends face toward the basal side. The mechanisms underlying this epithelial-specific microtubule assembly remain unresolved, however. Here, using mouse intestinal cells and human Caco-2 cells, we show that the microtubule minus-end binding protein CAMSAP3 (calmodulin-regulated-spectrin-associated protein 3) plays a pivotal role in orienting the apical-to-basal polarity of microtubules in epithelial cells. In these cells, CAMSAP3 accumulated at the apical cortices, and tethered the longitudinal microtubules to these sites. Camsap3 mutation or depletion resulted in a random orientation of these microtubules; concomitantly, the stereotypic positioning of the nucleus and Golgi apparatus was perturbed. In contrast, the integrity of the plasma membrane was hardly affected, although its structural stability was decreased. Further analysis revealed that the CC1 domain of CAMSAP3 is crucial for its apical localization, and that forced mislocalization of CAMSAP3 disturbs the epithelial architecture. These findings demonstrate that apically localized CAMSAP3 determines the proper orientation of microtubules, and in turn that of organelles, in mature mammalian epithelial cells. PMID:26715742

  12. Vaccination of sheep against haemonchosis with H11, a gut membrane-derived protective antigen from the adult parasite: prevention of the periparturient rise and colostral transfer of protective immunity.

    Science.gov (United States)

    Andrews, S J; Hole, N J; Munn, E A; Rolph, T P

    1995-07-01

    Pregnant ewes were immunised with a fraction highly enriched in the membrane glycoprotein antigen H11, isolated from the intestinal brush border of adult Haemonchus contortus. Immunity induced by immunisation was able to abolish almost completely (98-99%) the worm egg output from pregnant ewes challenged with ca. 10,000 infective larvae of H. contortus during the last trimester. Furthermore, lambs born and reared on vaccinated ewes had substantial antibody levels to H11 derived from maternal transfer. This antibody conferred moderate protection against a bolus challenge of ca. 3000 infective larvae of H. contortus in 5-week-old lambs.

  13. Apical dominance in Alstroemeria cultured in vitro

    NARCIS (Netherlands)

    Pumisutapon, P.; Visser, R.G.F.; Klerk, de G.J.M.

    2009-01-01

    Apical dominance in Alstroemeria is studied to develop an improved propagation protocol for this crop. Four types of explants were prepared: an intact rhizome with two intact shoots (+R+2S), an intact rhizome with two decapitated shoots (+R-2S), a decapitated rhizome with two intact shoots (-R+2S),

  14. Cell division activity during apical hook development

    NARCIS (Netherlands)

    Raz, V.; Koornneef, M.

    2001-01-01

    Growth during plant development is predominantly governed by the combined activities of cell division and cell elongation. The relative contribution of both activities controls the growth of a tissue. A fast change in growth is exhibited at the apical hypocotyl of etiolated seedlings where cells gro

  15. Genetic diversity of vaccine candidate antigens in Plasmodium falciparum isolates from the Amazon basin of Peru

    Directory of Open Access Journals (Sweden)

    Lucas Carmen M

    2008-05-01

    Full Text Available Abstract Background Several of the intended Plasmodium falciparum vaccine candidate antigens are highly polymorphic and could render a vaccine ineffective if their antigenic sites were not represented in the vaccine. In this study, characterization of genetic variability was performed in major B and T-cell epitopes within vaccine candidate antigens in isolates of P. falciparum from Peru. Methods DNA sequencing analysis was completed on 139 isolates of P. falciparum collected from endemic areas of the Amazon basin in Loreto, Peru from years 1998 to 2006. Genetic diversity was determined in immunological important regions in circumsporozoite protein (CSP, merozoite surface protein-1 (MSP-1, apical membrane antigen-1 (AMA-1, liver stage antigen-1 (LSA-1 and thrombospondin-related anonymous protein (TRAP. Alleles identified by DNA sequencing were aligned with the vaccine strain 3D7 and DNA polymorphism analysis and FST study-year pairwise comparisons were done using the DnaSP software. Multilocus analysis (MLA was performed and average of expected heterozygosity was calculated for each loci and haplotype over time. Results Three different alleles for CSP, seven for MSP-1 Block 2, one for MSP-1 Block 17, three for AMA-1 and for LSA-1 each and one for TRAP were identified. There were 24 different haplotypes in 125 infections with complete locus typing for each gene. Conclusion Characterization of the genetic diversity in Plasmodium isolates from the Amazon Region of Peru showed that P. falciparum T and B cell epitopes in these antigens have polymorphisms more similar to India than to Africa. These findings are helpful in the formulation of a vaccine considering restricted repertoire populations.

  16. Cleavage of endogenous γENaC and elevated abundance of αENaC are associated with increased Na+ transport in response to apical fluid volume expansion in human H441 airway epithelial cells

    OpenAIRE

    Tan, Chong D.; Selvanathar, Indusha A.; Baines, Deborah L.

    2011-01-01

    Using human H441 airway epithelial cells cultured at air–liquid interface (ALI), we have uniquely correlated the functional response to apical fluid volume expansion with the abundance and cleavage of endogenous α- and γENaC proteins in the apical membrane. Monolayers cultured at ALI rapidly elevated I sc when inserted into fluid-filled Ussing chambers. The increase in I sc was not significantly augmented by the apical addition of trypsin, and elevation was abolished by the protease inhibitor...

  17. Type III apical transportation of root canal

    Directory of Open Access Journals (Sweden)

    Shiv P Mantri

    2012-01-01

    Full Text Available Procedural accidents leading to complications such as canal transportation have been ascribed to inapt cleaning and shaping concepts. Canal transportation is an undesirable deviation from the natural canal path. Herewith a case of apical transportation of root canal resulting in endodontic retreatment failure and its management is presented. A healthy 21-year-old young male presented discomfort and swelling associated with painful endodontically retreated maxillary incisor. Radiograph revealed periradicular radiolucency involving underfilled 11 and overfilled 12. Insufficiently obturated 11 exhibited apical transportation of canal. This type III transportation was treated by periradicular surgery and repair using white mineral trioxide aggregate (MTA. Comfortable asymptomatic patient presented uneventful healing at third and fourth month recall visits. A decrease in the size of radiolucency in radiograph supported the clinical finding. In the present case, MTA is useful in repairing the transportation defect. The result of these procedures is predictable and successful.

  18. Shark Variable New Antigen Receptor (VNAR Single Domain Antibody Fragments: Stability and Diagnostic Applications

    Directory of Open Access Journals (Sweden)

    Stewart Nuttall

    2013-01-01

    Full Text Available The single variable new antigen receptor domain antibody fragments (VNARs derived from shark immunoglobulin new antigen receptor antibodies (IgNARs represent some of the smallest known immunoglobulin-based protein scaffolds. As single domains, they demonstrate favorable size and cryptic epitope recognition properties, making them attractive in diagnosis and therapy of numerous disease states. Here, we examine the stability of VNAR domains with a focus on a family of VNARs specific for apical membrane antigen 1 (AMA-1 from Plasmodium falciparum. The VNARs are compared to traditional monoclonal antibodies (mAbs in liquid, lyophilized and immobilized nitrocellulose formats. When maintained in various formats at 45 °C, VNARs have improved stability compared to mAbs for periods of up to four weeks. Using circular dichroism spectroscopy we demonstrate that VNAR domains are able to refold following heating to 80 °C. We also demonstrate that VNAR domains are stable during incubation under potential in vivo conditions such as stomach acid, but not to the protease rich environment of murine stomach scrapings. Taken together, our results demonstrate the suitability of shark VNAR domains for various diagnostic platforms and related applications.

  19. Light sheet microscopy for tracking single molecules on the apical surface of living cells.

    Science.gov (United States)

    Li, Yu; Hu, Ying; Cang, Hu

    2013-12-12

    Single particle tracking is a powerful tool to study single molecule dynamics in living biological samples. However, current tracking techniques, which are based mainly on epifluorescence, confocal, or TIRF microscopy, have difficulties in tracking single molecules on the apical surface of a cell. We present here a three-dimensional (3D) single particle tracking technique that is based on prism coupled light-sheet microscopy (PCLSM). This novel design provides a signal-to-noise ratio comparable to confocal microscopy while it has the capability of illuminating at arbitrary depth. We demonstrate tracking of single EGF molcules on the apical surface of live cell membranes from their binding to EGF receptors until they are internalized or photobleached. We found that EGF exhibits multiple diffusion behaviors on live A549 cell membranes. At room temperature, the average diffusion coefficient of EGF on A549 cells was measured to be 0.13 μm(2)/s. Depletion of cellular cholesterol with methyl-β-cyclodextrin leads to a broader distribution of diffusion coefficients and an increase of the average diffusion coefficient at room temperature. This light-sheet based 3D single particle tracking technique solves the technique difficulty of tracking single particles on apical membranes and is able to document the whole "lifetime" of a particle from binding till photobleaching or internalization. PMID:23895420

  20. Inflammatory Myofibroblastic Tumor Mimicking Apical Periodontitis.

    Science.gov (United States)

    Adachi, Makoto; Kiho, Kazuki; Sekine, Genta; Ohta, Takahisa; Matsubara, Makoto; Yoshida, Takakazu; Katsumata, Akitoshi; Tanuma, Jun-ichi; Sumitomo, Shinichiro

    2015-12-01

    Inflammatory myofibroblastic tumors (IMTs) are rare. IMTs of the head and neck occur in all age groups, from neonates to old age, with the highest incidence occurring in childhood and early adulthood. An IMT has been defined as a histologically distinctive lesion of uncertain behavior. This article describes an unusual case of IMT mimicking apical periodontitis in the mandible of a 42-year-old man. At first presentation, the patient showed spontaneous pain and percussion pain at teeth #28 to 30, which continued after initial endodontic treatment. Panoramic radiography revealed a radiolucent lesion at the site. Cone-beam computed tomographic imaging showed osteolytic lesions, suggesting an aggressive neoplasm requiring incisional biopsy. Histopathological examination indicated an IMT. The lesion was removed en bloc under general anesthesia, and the patient manifested no clinical evidence of recurrence for 24 months. Lesions of nonendodontic origin should be included in the differential diagnosis of apical periodontitis. Every available diagnostic tool should be used to confirm the diagnosis. Cone-beam computed tomographic imaging is very helpful for differential diagnosis in IMTs mimicking apical periodontitis. PMID:26602450

  1. B-cell responses to pregnancy-restricted and -unrestricted Plasmodium falciparum erythrocyte membrane protein 1 antigens in Ghanaian women naturally exposed to malaria parasites

    DEFF Research Database (Denmark)

    Ampomah, Paulina; Stevenson, Liz; Ofori, Michael F;

    2014-01-01

    Protective immunity to Plasmodium falciparum malaria acquired after natural exposure is largely antibody mediated. IgG-specific P. falciparum EMP1 (PfEMP1) proteins on the infected erythrocyte surface are particularly important. The transient antibody responses and the slowly acquired protective...... immunity probably reflect the clonal antigenic variation and allelic polymorphism of PfEMP1. However, it is likely that other immune-evasive mechanisms are also involved, such as interference with formation and maintenance of immunological memory. We measured PfEMP1-specific antibody levels by enzyme......-linked immunosorbent assay (ELISA) and memory B-cell frequencies by enzyme-linked immunosorbent spot (ELISPOT) assay in a cohort of P. falciparum-exposed nonpregnant Ghanaian women. The antigens used were a VAR2CSA-type PfEMP1 (IT4VAR04) with expression restricted to parasites infecting the placenta, as well as two...

  2. 5-Azacytidine up regulates the expression of Epstein-Barr virus nuclear antigen 2 (EBNA-2) through EBNA-6 and latent membrane protein in the Burkitt's lymphoma line rael.

    Science.gov (United States)

    Masucci, M G; Contreras-Salazar, B; Ragnar, E; Falk, K; Minarovits, J; Ernberg, I; Klein, G

    1989-01-01

    Nonproductive infection of B lymphocytes by Epstein-Barr virus (EBV) is associated with a highly restricted expression of viral genes. In growth-transformed lymphoblastoid cell lines, the products of these genes include a complex of at least six EBV nuclear antigens (EBNAs) (EBNA-1 through EBNA-6) and one membrane protein (latent membrane protein [LMP]). EBV-carrying Burkitt's lymphoma (BL) biopsies and derived cell lines that have retained a representative phenotype (group I BL lines) express only EBNA-1 (M. Rowe, D. T. Rowe, C. D. Gregory, L. S. Young, P. J. Farrell, H. Rupani, and A. B. Rickinson, EMBO J. 6:2743-2751, 1987). We have found that EBNA-2 through EBNA-6 and LMP can be up regulated by treating the group I BL line Rael with the DNA-demethylating agent 5-azacytidine (5-AzaC). The drug acted in a time- and dose-dependent manner. EBNA-2-positive cells were detected by anti-complement immunofluorescence staining just 12 h after addition of 4 microM 5-AzaC and reached a maximum number at 72 h, when up to 75% of the population was positive. EBNA-2, EBNA-3, EBNA-4, EBNA-4, EBNA-6, and LMP were demonstrated immunoblots starting at 48 h. The EBV-encoded early antigens and viral capsid antigens were also induced but at a lower level. EBNA-2 and the lytic cycle-associated antigens appeared with a different time course and in largely nonoverlapping cell subpopulations, as demonstrated by double fluorescence staining. Thus, EBNA-2 expression was not restricted to lytically infected cells, nor was EBNA-2 required for entry into the lytic cycle. The coding and regulatory sequences of EBNA-2 and LMP were found to be highly methylated in Rael cells and were, as expected, demethylated after 5-AzaC treatment. These findings suggest that DNA methylation may participate in the regulation of growth transformation-associated viral genes in BL cells. Images PMID:2470924

  3. Uteroglobin, an apically secreted protein of the uterine epithelium, is secreted non-polarized form MDCK cells and mainly basolaterally from Caco-2 cells

    DEFF Research Database (Denmark)

    Vogel, L K; Suske, G; Beato, M;

    1993-01-01

    A complete cDNA encoding rabbit uteroglobin was constructed and expressed in MDCK and Caco-2 cells. The MDCK cells secrete uteroglobin in approximately equal amounts to the apical and the basolateral side, whereas the Caco-2 cells secrete uteroglobin mainly to the basolateral side. Both MDCK and ...... the endometrial epithelium has an apical default pathway or recognises a sorting signal not recognised by MDCK cells and Caco-2 cells. Our data thus show that a soluble molecule can be secreted at the apical, the basolateral or both membranes depending on the cell type....

  4. Functional Apical Large Conductance, Ca2+-activated, and Voltage-dependent K+ Channels Are Required for Maintenance of Airway Surface Liquid Volume*

    OpenAIRE

    Manzanares, Dahis; Gonzalez, Carlos; Ivonnet, Pedro; Chen, Ren-Shiang; Valencia-Gattas, Monica; Gregory E. Conner; Larsson, H. Peter; Salathe, Matthias

    2011-01-01

    Large conductance, Ca2+-activated, and voltage-dependent K+ (BK) channels control a variety of physiological processes in nervous, muscular, and renal epithelial tissues. In bronchial airway epithelia, extracellular ATP-mediated, apical increases in intracellular Ca2+ are important signals for ion movement through the apical membrane and regulation of water secretion. Although other, mainly basolaterally expressed K+ channels are recognized as modulators of ion transport in airway epithelial ...

  5. Human bladder cancer stem cells exist in epithelial membrane antigen-subset%人膀胱癌干细胞存在于EMA-细胞亚群

    Institute of Scientific and Technical Information of China (English)

    杨宇明; 畅继武

    2008-01-01

    BACKGROUND:Cancer stem cell (CSC) hypothesis suggests that tumorous clones are maintained by a rare fraction of cells with stem cell proprieties. Several kinds of CSCs of solid tumor have been isolated in recent years. However, there have been fewer studies on the objective existence of bladder cancer stem cells (BCSCs) and on the methods to effectively isolate and identify BCSCs. OBJECTIVE:To investigate possibilities of BCSC existence and of epithelial membrane antigen (EMA) used as a surface marker of BCSC. DESIGN:A control observation experiment. SETTING:Tianjin Institute of Urinary Surgery & Second Hospital of Tianjin Medical University. MATERIALS:This study was performed at the Room for Tumor Immunity of Tianjin Institute of Urinary Surgery (key laboratory for State "211 Project") from March 2006 to July 2007. Nine specimens of human bladder were obtained from patients who received treatment in the Second Hospital of Tianjin Medical University. These specimens corresponded to the diagnostic criteria of low malignant potential papillary urothelial neoplasm and low-grade papillary urothelial carcinoma. Additionally, 40 samples of human low malignant bladder transitional cell carcinomas (BTCC) and 10 samples of normal urothelium that were used for immunohistochemistry were obtained from the patients who received treatment in the Department of Urinary Surgery, Second Hospital of Tianjin Medical University. Written informed consent for the specimen providing was obtained from the patients, and the protocol was approved by the hospital’s Ethics Committee. METHODS:The genes that were differentially expressed between normal urothelium and BTCC were identified through a DNA array assay to preliminarily determine the existence of BTCC. Overpressed stem cell related genes, Bmi-1 and EZH2, were verified by immunohistochemistry. A total of 27 potential surface markers of BCSCs were assayed to determine the location of positive cells. EMA- subsets were obtained through

  6. Evidence for an apical Na-Cl cotransporter involved in ion uptake in a teleost fish

    Science.gov (United States)

    Hiroi, J.; Yasumasu, S.; McCormick, S.D.; Hwang, P.-P.; Kaneko, T.

    2008-01-01

    Cation-chloride cotransporters, such as the Na+/K +/2Cl- cotransporter (NKCC) and Na+/Cl - cotransporter (NCC), are localized to the apical or basolateral plasma membranes of epithelial cells and are involved in active ion absorption or secretion. The objectives of this study were to clone and identify 'freshwater-type' and 'seawater-type' cation-chloride cotransporters of euryhaline Mozambique tilapia (Oreochromis mossambicus) and to determine their intracellular localization patterns within mitochondria-rich cells (MRCs). From tilapia gills, we cloned four full-length cDNAs homologous to human cation-chloride cotransporters and designated them as tilapia NKCC1a, NKCC1b, NKCC2 and NCC. Out of the four candidates, the mRNA encoding NKCC1a was highly expressed in the yolk-sac membrane and gills (sites of the MRC localization) of seawater-acclimatized fish, whereas the mRNA encoding NCC was exclusively expressed in the yolk-sac membrane and gills of freshwater-acclimatized fish. We then generated antibodies specific for tilapia NKCC1a and NCC and conducted whole-mount immunofluorescence staining for NKCC1a and NCC, together with Na+/K+-ATPase, cystic fibrosis transmembrane conductance regulator (CFTR) and Na+/H+ exchanger 3 (NHE3), on the yolk-sac membrane of tilapia embryos acclimatized to freshwater or seawater. The simultaneous quintuple-color immunofluorescence staining allowed us to classify MRCs clearly into four types: types I, II, III and IV. The NKCC1a immunoreactivity was localized to the basolateral membrane of seawater-specific type-IV MRCs, whereas the NCC immunoreactivity was restricted to the apical membrane of freshwater-specific type-II MRCs. Taking account of these data at the level of both mRNA and protein, we deduce that NKCC1a is the seawater-type cotransporter involved in ion secretion by type-IV MRCs and that NCC is the freshwater-type cotransporter involved in ion absorption by type-II MRCs. We propose a novel ion-uptake model by MRCs in

  7. Apical dominance in Alstroemeria cultured in vitro

    OpenAIRE

    Pumisutapon, P.; Visser, R. G. F.; Klerk, de, L.A.

    2009-01-01

    Apical dominance in Alstroemeria is studied to develop an improved propagation protocol for this crop. Four types of explants were prepared: an intact rhizome with two intact shoots (+R+2S), an intact rhizome with two decapitated shoots (+R-2S), a decapitated rhizome with two intact shoots (-R+2S), and a decapitated rhizome with two decapitated shoots (-R-2S). The explants were cultivated on solid MS medium with 9 µM 6-benzylaminopurine (BAP). -R-2S explants showed the highest and +R+2S the l...

  8. Multimodality imaging in apical hypertrophic cardiomyopathy

    Institute of Scientific and Technical Information of China (English)

    Rosario; Parisi; Francesca; Mirabella; Gioel; Gabrio; Secco; Rossella; Fattori

    2014-01-01

    Apical hypertrophic cardiomyopathy(AHCM) is a relatively rare morphologic variant of HCM in which the hypertrophy of myocardium is localized to the left ventricular apex. Symptoms of AHCM might vary from none to others mimic coronary artery disease including acute coronary syndrome, thus resulting in inappropriate hospitalization. Transthoracic echocardiography is the firstline imaging technique for the diagnosis of hypertrophic cardiomyopathies. However, when the hypertrophy of the myocardium is localized in the ventricular apex might results in missed diagnosis. Aim of this paper is to review the different imaging techniques used for the diagnosis of AHCM and their role in the detection and comprehension of this uncommon disease.

  9. 前列腺特异性膜抗原为靶标的放射免疫治疗进展%Advances in radioimmunotherapy targeting prostate speciifc membrane antigen in prostate cancer

    Institute of Scientific and Technical Information of China (English)

    刘冲(综述); 陶嵘(审校)

    2015-01-01

    前列腺特异性膜抗原(PSMA)是一种跨膜糖蛋白,几乎表达于所有前列腺癌,在转移性激素抵抗性前列腺癌中表达量显著增加。放射免疫治疗(RIT)利用放射性核素标记单克隆抗体以实现肿瘤的靶向治疗。本文对PSMA为靶标的RIT进展作一综述。%Prostate specific membrane antigen (PSMA) is a transmembrane glycoprotein, which is almost expressed in all prostate cancers. The expression of PSMA in metastatic hormone-refractory prostate cancer is significantly increased. Radioimmunotherapy combined radionuclide with monoclonal antibody is a kind of targeted cancer therapy. This review will summarize the progress of radioimmunotherapy targeting PSMA.

  10. ANTI-NUCLEOSOME ANTIBODIES COMPLEXED TO NUCLEOSOMAL ANTIGENS SHOW ANTI-DNA REACTIVITY AND BIND TO RAT GLOMERULAR-BASEMENT-MEMBRANE IN-VIVO

    NARCIS (Netherlands)

    KRAMERS, C; HYLKEMA, MN; VANBRUGGEN, MCJ; VANDELAGEMAAT, R; DIJKMAN, HBPM; ASSMANN, KJM; SMEENK, RJT; BERDEN, JHM; Hylkema, Machteld

    1994-01-01

    Histones can mediate the binding of DNA and anti-DNA to the glomerular basement membrane (GBM). Zn ELISA histone/DNA/anti-DNA complexes are able to bind to heparan sulfate (HS), an intrinsic constituent of the GBM. We questioned whether histone containing immune complexes are able to bind to the GBM

  11. The expression and functional activity of membrane-bound human leukocyte antigen-G1 are influenced by the 3'-untranslated region

    DEFF Research Database (Denmark)

    Svendsen, Signe Goul; Hantash, Basil M; Zhao, Longmei;

    2013-01-01

    , and graft rejection after organ transplantation. We have investigated the effect of HLA-G polymorphism in the 3'UTR on the processing and stability of the membrane-bound HLA-G1 (mHLA-G1) isoform, as well as its functional significance. Different HLA-G1 cDNA sequences were transduced into the human K562 cell...

  12. Tetrahymena gene encodes a protein that is homologous with the liver-specific F-antigen and associated with membranes of the Golgi apparatus and transport vesicles

    DEFF Research Database (Denmark)

    Hummel, R; Nørgaard, P; Andreasen, P H;

    1992-01-01

    of the Golgi apparatus and transport vesicles pointing to a role of TF-ag in membrane trafficking. Transcription of the TF-ag gene, as determined by run-on analyses, was only detectable in growing cells, and following transfer to starvation condition pre-existing TF-ag mRNA was rapidly degraded. The abundance...

  13. Contribution of cubilin and amnionless to processing and membrane targeting of cubilin-amnionless complex

    DEFF Research Database (Denmark)

    Coudroy, Gwénaëlle; Gburek, Jakub; Kozyraki, Renata;

    2005-01-01

    fragments, including a functional "mini" version of cubilin, the processing, sorting, and membrane anchoring of the complex to the apical membrane were investigated. The results show that truncation mutants, including the N-terminal domain of cubilin, did not appear at the plasma membrane but instead were...... cubilin at the apical cell surface. Apical sorting was observed for a broad set of nonoverlapping cubilin fragments without the N-terminal region, in the absence of AMN. The preference for apical sorting disappeared when glycosylation was inhibited by tunicamycin. In conclusion, it is shown that both...

  14. Apical dominance and growth in vitro of Alstroemeria

    OpenAIRE

    Pumisutapon, P.

    2012-01-01

    In Alstroemeria, micropropagation is achieved by axillary bud outgrowth. However, the multiplication rate is rather low (1.2–2.0 per cycle of 4 weeks) due to strong apical dominance. Even though several factors (i.e. culture media, growth regulators, and environmental conditions) have been studied, no significant improvements have been achieved. Basic research on apical dominance mechanism in Alstroemeria is therefore required. This may enhance the understanding how apical dominance con...

  15. Oncostatin M regulates membrane traffic and stimulates bile canalicular membrane biogenesis in HepG2 cells

    NARCIS (Netherlands)

    Van der Wouden, Johanna M.; Van IJzendoorn, Sven C.D.; Hoekstra, Dick

    2002-01-01

    Hepatocytes are the major epithelial cells of the liver and they display membrane polarity: the sinusoidal membrane representing the basolateral surface, while the bile canalicular membrane is typical of the apical membrane. In polarized HepG2 cells an endosomal organelle, SAC, fulfills a prominent

  16. Modulatory Effect of Taurine on 7,12-Dimethylbenz(a)Anthracene-Induced Alterations in Detoxification Enzyme System, Membrane Bound Enzymes, Glycoprotein Profile and Proliferative Cell Nuclear Antigen in Rat Breast Tissue.

    Science.gov (United States)

    Vanitha, Manickam Kalappan; Baskaran, Kuppusamy; Periyasamy, Kuppusamy; Selvaraj, Sundaramoorthy; Ilakkia, Aruldoss; Saravanan, Dhiravidamani; Venkateswari, Ramachandran; Revathi Mani, Balasundaram; Anandakumar, Pandi; Sakthisekaran, Dhanapal

    2016-08-01

    The modulatory effect of taurine on 7,12-dimethylbenz(a)anthracene (DMBA)-induced breast cancer in rats was studied. DMBA (25 mg/kg body weight) was administered to induce breast cancer in rats. Protein carbonyl levels, activities of membrane bound enzymes (Na(+) /K(+) ATPase, Ca(2+) ATPase, and Mg(2+) ATPase), phase I drug metabolizing enzymes (cytochrome P450, cytochrome b5, NADPH cytochrome c reductase), phase II drug metabolizing enzymes (glutathione-S-transferase and UDP-glucuronyl transferase), glycoprotein levels, and proliferative cell nuclear antigen (PCNA) were studied. DMBA-induced breast tumor bearing rats showed abnormal alterations in the levels of protein carbonyls, activities of membrane bound enzymes, drug metabolizing enzymes, glycoprotein levels, and PCNA protein expression levels. Taurine treatment (100 mg/kg body weight) appreciably counteracted all the above changes induced by DMBA. Histological examination of breast tissue further supported our biochemical findings. The results of the present study clearly demonstrated the chemotherapeutic effect of taurine in DMBA-induced breast cancer. PMID:27091720

  17. Modulatory Effect of Taurine on 7,12-Dimethylbenz(a)Anthracene-Induced Alterations in Detoxification Enzyme System, Membrane Bound Enzymes, Glycoprotein Profile and Proliferative Cell Nuclear Antigen in Rat Breast Tissue.

    Science.gov (United States)

    Vanitha, Manickam Kalappan; Baskaran, Kuppusamy; Periyasamy, Kuppusamy; Selvaraj, Sundaramoorthy; Ilakkia, Aruldoss; Saravanan, Dhiravidamani; Venkateswari, Ramachandran; Revathi Mani, Balasundaram; Anandakumar, Pandi; Sakthisekaran, Dhanapal

    2016-08-01

    The modulatory effect of taurine on 7,12-dimethylbenz(a)anthracene (DMBA)-induced breast cancer in rats was studied. DMBA (25 mg/kg body weight) was administered to induce breast cancer in rats. Protein carbonyl levels, activities of membrane bound enzymes (Na(+) /K(+) ATPase, Ca(2+) ATPase, and Mg(2+) ATPase), phase I drug metabolizing enzymes (cytochrome P450, cytochrome b5, NADPH cytochrome c reductase), phase II drug metabolizing enzymes (glutathione-S-transferase and UDP-glucuronyl transferase), glycoprotein levels, and proliferative cell nuclear antigen (PCNA) were studied. DMBA-induced breast tumor bearing rats showed abnormal alterations in the levels of protein carbonyls, activities of membrane bound enzymes, drug metabolizing enzymes, glycoprotein levels, and PCNA protein expression levels. Taurine treatment (100 mg/kg body weight) appreciably counteracted all the above changes induced by DMBA. Histological examination of breast tissue further supported our biochemical findings. The results of the present study clearly demonstrated the chemotherapeutic effect of taurine in DMBA-induced breast cancer.

  18. Minimal Apical Enlargement for Penetration of Irrigants to the Apical Third of Root Canal System: A Scanning Electron Microscope Study

    OpenAIRE

    P. Srikanth; Krishna, Amaravadi Gopi; Srinivas, Siva; Reddy, E Sujayeendranatha; Battu, Someshwar; Aravelli, Swathi

    2015-01-01

    Background: The aim of this study was to determine minimal apical enlargement for irrigant penetration into apical third of root canal system using scanning electron microscope (SEM). Materials and Methods: Distobuccal canals of 40 freshly extracted human maxillary first molar teeth were instrumented using crown-down technique. The teeth were divided into four test groups according to size of their master apical file (MAF) (#20, #25, #30, #35 0.06% taper), and two control groups. After final ...

  19. Loss of primary cilia results in deregulated and unabated apical calcium entry in ARPKD collecting duct cells.

    Science.gov (United States)

    Siroky, Brian J; Ferguson, William B; Fuson, Amanda L; Xie, Yi; Fintha, Attila; Komlosi, Peter; Yoder, Bradley K; Schwiebert, Erik M; Guay-Woodford, Lisa M; Bell, P Darwin

    2006-06-01

    Recent genetic analysis has identified a pivotal role of primary cilia in the pathogenesis of polycystic kidney disease (PKD). However, little is known regarding how cilia loss/dysfunction contributes to cyst development. In epithelial cells, changes in apical fluid flow induce cilia-mediated Ca2+ entry via polycystin-2 (PC2), a cation channel. The Oak Ridge Polycystic Kidney (orpk) mouse contains a mutated Tg737 gene that disrupts expression of polaris, a protein required for ciliogenesis. These studies examine the effect of cilia malformation on Ca2+ entry in orpk cilia(-) collecting duct principal cells, and in orpk cells in which wild-type Tg737 was reintroduced, orpk cilia(+). [Ca2+]i was monitored in confluent cell monolayers using fluorescence microscopy. Intrinsic apical Ca2+ entry was measured by Mn2+ quenching and Ca2+ depletion/readdition under flow conditions below the threshold for stimulation. We found that unstimulated apical Ca2+ entry was markedly increased in cilia(-) cells and was sensitive to Gd3+, an inhibitor of PC2. Electrophysiological measurements demonstrate increased abundance of an apical channel, consistent with PC2, in cilia(-) cells. Immunofluorescence studies revealed that PC2, normally expressed on and at the base of cilia in orpk cilia(+) cells, was observed throughout the apical membrane in cilia(-) cells. Furthermore, cilia(-) cells displayed elevated subapical Ca2+ levels measured with the near-membrane Ca2+ indicator FFP-18. We propose that cilia exert a tonic regulatory influence on apical Ca2+ entry, and absence of cilia results in loss of spatial organization of PC2, causing unregulated Ca2+ entry and elevations in subapical [Ca2+], a factor which may contribute to cyst formation. PMID:16396941

  20. Within-population genetic diversity of Plasmodium falciparum vaccine candidate antigens reveals geographic distance from a Central sub-Saharan African origin.

    Science.gov (United States)

    Tanabe, Kazuyuki; Mita, Toshihiro; Palacpac, Nirianne M Q; Arisue, Nobuko; Tougan, Takahiro; Kawai, Satoru; Jombart, Thibaut; Kobayashi, Fumie; Horii, Toshihiro

    2013-02-18

    Populations of Plasmodium falciparum, the most virulent human malaria parasite, are diverse owing to wide levels of transmission and endemicity of infection. Genetic diversity of P. falciparum antigens, within and between parasite populations, remains a confounding factor in malaria pathogenesis as well as clinical trials of vaccine candidates. Variation of target antigens in parasite populations may arise from immune pressure depending on the levels of acquired immunity. Alternatively, similar to our study in housekeeping genes [Tanabe et al. Curr Biol 2010;70:1-7], within-population genetic diversity of vaccine candidate antigens may also be determined by geographical distance from a postulated origin in Central sub-Saharan Africa. To address this question, we obtained full-length sequences of P. falciparum genes, apical membrane antigen 1 (ama1) (n=459), circumsporozoite protein (csp) (n=472) and merozoite surface protein 1 (msp1) (n=389) from seven geographically diverse parasite populations in Africa, Southeast Asia and Oceania; and, together with previously determined sequences (n=13 and 15 for csp and msp1, respectively) analyzed within-population single nucleotide polymorphism (SNP) diversity. The three antigen genes showed SNP diversity that supports a model of isolation-by-distance. The standardized number of polymorphic sites per site, expressed as θ(S), indicates that 77-83% can be attributed by geographic distance from the African origin, suggesting that geographic distance plays a significant role in variation in target vaccine candidate antigens. Furthermore, we observed that a large proportion of SNPs in the antigen genes were shared between African and non-African parasite populations, demonstrating long term persistence of those SNPs. Our results provide important implications for developing effective malaria vaccines and better understanding of acquired immunity against falciparum malaria. PMID:23295064

  1. APICAL TUBERCULOSIS PRESENTING AS PANCOAST TUMOR

    Directory of Open Access Journals (Sweden)

    Praveen

    2014-07-01

    Full Text Available A 73-year-old woman presented with pain in her right shoulder radiating to the right scapula and a tingling sensation of the right arm with involvement of the fourth and fifth finger. Chest x ray showed a well-defined mass in right apical area of lung. HRCT chest showed a well-defined mass in right upper zone in apex destroying the upper 2 ribs. Clinically the diagnosis of Pancoast tumor of the right lung was made. Computed tomographic guided fine needle aspiration cytology was done and it was inconclusive. So gun biopsy was done under ultrasound guidance. The histologic and microbiologic examinations established the diagnosis of tuberculosis (TB.

  2. Apical-dominant particle swarm optimization

    Institute of Scientific and Technical Information of China (English)

    Zhihua Cui; Xingjuan Cai; Jianchao Zeng; Guoji Sun

    2008-01-01

    Particle swarm optimization (PSO) is a new stochastic population-based search methodology by simulating the animal social behaviors such as birds flocking and fish schooling.Many improvements have been proposed within the framework of this biological assumption.However,in this paper,the search pattern of PSO is used to model the branch growth process of natural plants.It provides a different poten-tial manner from artificial plant.To illustrate the effectiveness of this new model,apical dominance phenomenon is introduced to construct a novel variant by emphasizing the influence of the phototaxis.In this improvement,the population is divided into three different kinds of buds associated with their performances.Furthermore,a mutation strategy is applied to enhance the ability escaping from a local optimum.Sim-ulation results demonstrate good performance of the new method when solving high-dimensional multi-modal problems.

  3. Loss of apical monocilia on collecting duct principal cells impairs ATP secretion across the apical cell surface and ATP-dependent and flow-induced calcium signals.

    Science.gov (United States)

    Hovater, Michael B; Olteanu, Dragos; Hanson, Elizabeth L; Cheng, Nai-Lin; Siroky, Brian; Fintha, Attila; Komlosi, Peter; Liu, Wen; Satlin, Lisa M; Bell, P Darwin; Yoder, Bradley K; Schwiebert, Erik M

    2008-06-01

    mechanical pipetting stimuli trigger release of a common ATP pool. Cilium-competent monolayers responded to flow with an increase in cell Ca(2+) derived from both extracellular and intracellular stores. This flow-induced Ca(2+) signal was less robust in cilium-deficient monolayers. Flow-induced Ca(2+) signals in both preparations were attenuated by extracellular gadolinium and by extracellular apyrase, an ATPase/ADPase. Taken together, these data suggest that apical monocilia are sensory organelles and that their presence in the apical membrane facilitates the formation of a mature ATP secretion apparatus responsive to chemical, osmotic, and mechanical stimuli. The cilium and autocrine ATP signaling appear to work in concert to control cell Ca(2+). Loss of a cilium-dedicated autocrine purinergic signaling system may be a critical underlying etiology for ARPKD and may lead to disinhibition and/or upregulation of multiple sodium (Na(+)) absorptive mechanisms and a resultant severe hypertensive phenotype in ARPKD and, possibly, other diseases. PMID:18368523

  4. Stable solid-phase Rh antigen.

    Science.gov (United States)

    Yared, M A; Moise, K J; Rodkey, L S

    1997-12-01

    Numerous investigators have attempted to isolate the Rh antigens in a stable, immunologically reactive form since the discovery of the Rh system over 56 years ago. We report here a successful and reproducible approach to solubilizing and adsorbing the human Rh antigen(s) to a solid-phase matrix in an antigenically active form. Similar results were obtained with rabbit A/D/F red blood cell antigens. The antigen preparation was made by dissolution of the red blood cell membrane lipid followed by fragmentation of the residual cytoskeleton in an EDTA solution at low ionic strength. The antigenic activity of the soluble preparations was labile in standard buffers but was stable in zwitterionic buffers for extended periods of time. Further studies showed that the antigenic activity of these preparations was enhanced, as was their affinity for plastic surfaces, in the presence of acidic zwitterionic buffers. Adherence to plastic surfaces at low pH maintained antigenic reactivity and specificity for antibody was retained. The data show that this approach yields a stable form of antigenically active human Rh D antigen that could be used in a red blood cell-free assay for quantitative analysis of Rh D antibody and for Rh D antibody immunoadsorption and purification.

  5. Reversible transient apical ballooning syndrome with coronary lesions

    Institute of Scientific and Technical Information of China (English)

    Yunshan Cao; Min Zhang; Xiang Li; Ping Xie; Lynn Cronin

    2009-01-01

    Transient apical ballooning syndrome(Tako-Tsubo syndrome or ampulla cardiomyopathy) occurs predominantly in women over 60 years of age with a history of recent physical or psychological stress. We present a case of a male patient with reversible transient apical ballooning syndrome with significant coronary lesions and other ECG changes that did not explain the clinical symptoms.

  6. Unique apicomplexan IMC sub-compartment proteins are early markers for apical polarity in the malaria parasite.

    Science.gov (United States)

    Poulin, Benoit; Patzewitz, Eva-Maria; Brady, Declan; Silvie, Olivier; Wright, Megan H; Ferguson, David J P; Wall, Richard J; Whipple, Sarah; Guttery, David S; Tate, Edward W; Wickstead, Bill; Holder, Anthony A; Tewari, Rita

    2013-01-01

    The phylum Apicomplexa comprises over 5000 intracellular protozoan parasites, including Plasmodium and Toxoplasma, that are clinically important pathogens affecting humans and livestock. Malaria parasites belonging to the genus Plasmodium possess a pellicle comprised of a plasmalemma and inner membrane complex (IMC), which is implicated in parasite motility and invasion. Using live cell imaging and reverse genetics in the rodent malaria model P. berghei, we localise two unique IMC sub-compartment proteins (ISPs) and examine their role in defining apical polarity during zygote (ookinete) development. We show that these proteins localise to the anterior apical end of the parasite where IMC organisation is initiated, and are expressed at all developmental stages, especially those that are invasive. Both ISP proteins are N-myristoylated, phosphorylated and membrane-bound. Gene disruption studies suggest that ISP1 is likely essential for parasite development, whereas ISP3 is not. However, an absence of ISP3 alters the apical localisation of ISP1 in all invasive stages including ookinetes and sporozoites, suggesting a coordinated function for these proteins in the organisation of apical polarity in the parasite.

  7. Unique apicomplexan IMC sub-compartment proteins are early markers for apical polarity in the malaria parasite

    Directory of Open Access Journals (Sweden)

    Benoit Poulin

    2013-09-01

    The phylum Apicomplexa comprises over 5000 intracellular protozoan parasites, including Plasmodium and Toxoplasma, that are clinically important pathogens affecting humans and livestock. Malaria parasites belonging to the genus Plasmodium possess a pellicle comprised of a plasmalemma and inner membrane complex (IMC, which is implicated in parasite motility and invasion. Using live cell imaging and reverse genetics in the rodent malaria model P. berghei, we localise two unique IMC sub-compartment proteins (ISPs and examine their role in defining apical polarity during zygote (ookinete development. We show that these proteins localise to the anterior apical end of the parasite where IMC organisation is initiated, and are expressed at all developmental stages, especially those that are invasive. Both ISP proteins are N-myristoylated, phosphorylated and membrane-bound. Gene disruption studies suggest that ISP1 is likely essential for parasite development, whereas ISP3 is not. However, an absence of ISP3 alters the apical localisation of ISP1 in all invasive stages including ookinetes and sporozoites, suggesting a coordinated function for these proteins in the organisation of apical polarity in the parasite.

  8. Structural Studies on Plasmodium falciparum Erythrocyte Membrane Protein 1 (PfEMP1) Malaria Antigens Using Small Angle X-Ray Scattering (SAXS)

    DEFF Research Database (Denmark)

    Christoffersen, Stig

    Infection with the pathogenic Plasmodium falciparum parasite causes the potentially deadly Malaria disease which leads to over 1 million fatalities each year according to the WHO (World Health Organization). Individuals subjected to multiple infections gradually become immune to the disease sympt...... covers many aspects of PfEMP1 structural composition and mechanism. Information that could be of value to future vaccine developments which ultimately will prevent the severe and fatal outcomes that malaria afflicts.......Infection with the pathogenic Plasmodium falciparum parasite causes the potentially deadly Malaria disease which leads to over 1 million fatalities each year according to the WHO (World Health Organization). Individuals subjected to multiple infections gradually become immune to the disease...... for low-resolution structural characterization of the PfEMP1 proteins. This thesis describes how small angle X-ray scattering (SAXS) was used to achieve low-resolution structural information for a range of PfEMP1 proteins investigated as potent malaria antigens. Especially the leading vaccine candidate...

  9. Genetic interaction of centrosomin and bazooka in apical domain regulation in Drosophila photoreceptor.

    Directory of Open Access Journals (Sweden)

    Geng Chen

    Full Text Available BACKGROUND: Cell polarity genes including Crumbs (Crb and Par complexes are essential for controlling photoreceptor morphogenesis. Among the Crb and Par complexes, Bazooka (Baz, Par-3 homolog acts as a nodal component for other cell polarity proteins. Therefore, finding other genes interacting with Baz will help us to understand the cell polarity genes' role in photoreceptor morphogenesis. METHODOLOGY/PRINCIPAL FINDINGS: Here, we have found a genetic interaction between baz and centrosomin (cnn. Cnn is a core protein for centrosome which is a major microtubule-organizing center. We analyzed the effect of the cnn mutation on developing eyes to determine its role in photoreceptor morphogenesis. We found that Cnn is dispensable for retinal differentiation in eye imaginal discs during the larval stage. However, photoreceptors deficient in Cnn display dramatic morphogenesis defects including the mislocalization of Crumbs (Crb and Bazooka (Baz during mid-stage pupal eye development, suggesting that Cnn is specifically required for photoreceptor morphogenesis during pupal eye development. This role of Cnn in apical domain modulation was further supported by Cnn's gain-of-function phenotype. Cnn overexpression in photoreceptors caused the expansion of the apical Crb membrane domain, Baz and adherens junctions (AJs. CONCLUSIONS/SIGNIFICANCE: These results strongly suggest that the interaction of Baz and Cnn is essential for apical domain and AJ modulation during photoreceptor morphogenesis, but not for the initial photoreceptor differentiation in the Drosophila photoreceptor.

  10. Phosphatidylserine-positive particles in the apical domain of sensory hair cells

    Institute of Scientific and Technical Information of China (English)

    SHI Xiao-rui; Alfred Nuttall

    2006-01-01

    Apical membrane recycling has been proposed to be important for normal hair cell function. The current study reports an in vitro work that demonstrates the presence of phosphatidylserine (PS) and PS-positive vesicles labeled by Annexin V in the apical portion of hair cells. The following characteristics of the PS-positive vesicles were noticed using scanning confocal fluorescence microscopy: (1) variable sizes around 200 nm; (2)variable distribution patterns (either uniformly along individual stereocilia in the hair bundle or irregular) in the stereocilia from cell to cell; (3) variable sizes and numbers at locations along the border of the cuticular plate (CP),with a large number of them located at the vestigal kinocilial location; (4) motility with some of the vesicles during the observation period; (5) increase in PS labeling and the number of PS-positive vesicles after loud sound stimulation; and (6) decreased PS labeling and PS-positive vesicle numbers following treatment with LY-294002, a PI3 -kinase inhibitor. These results suggest that the presence of PS-positive vesicles at the apical area of hair cells may be indicative of vesicle shedding or transportation of a protein or rafts.

  11. Correlative Analysis of [Ca2+]C and Apical Secretion during Pollen Tube Growth and Reorientation

    OpenAIRE

    Coelho, Pedro Castanho; Malhó, Rui

    2006-01-01

    The maintenance of a cytosolic free calcium gradient (Ca2+]c) and vesicle secretion in the apex of pollen tubes is essential for growth. It has been postulated that high [Ca2+]c levels promote and confine vesicle fusion with the apical plasma membrane and in this study we performed a correlative analysis of both events using specific fluorescent dyes and confocal scanning microscopy. [Ca2+]c was imaged with Calcium Green-1 10 kDa dextran (CG-1) while secretory events were followed with FM1–43...

  12. The antimicrobial effect of apical box versus apical cone preparation using iodine potassium iodide as root canal dressing

    DEFF Research Database (Denmark)

    Markvart, Merete; Dahlén, Gunnar; Reit, Claes-Erik;

    2013-01-01

    Abstract Purpose. The purpose was to study the reduction of intra-canal microflora in premolars with apical periodontitis instrumented with either apical box or apical cone preparation and to provide measurements of intervention effects to allow proper power calculation in future clinical trials....... Methods. Twenty-four patients were centrally randomized to apical box preparation (size #60) or cone preparation (apical size #25). The groups were comparable regarding the presence of primary caries and type of coronal restoration. In the course of canal preparation each tooth was irrigated with 2.5% Na......OCl (12 ml). Lastly, the canals were filled with 17% EDTA (2 × 30 s) and 5% iodine potassium iodide (IKI) for 10 min. The canals were sampled for micro-organisms on four occasions: before instrumentation, after instrumentation, after application of IKI dressing and at the beginning of the second...

  13. Maxillary first premolars: I. Morphology of the apical constriction

    Institute of Scientific and Technical Information of China (English)

    Daming Wu; Younong Wu; Ming Hu

    2009-01-01

    Objective: To investigated the apical constriction morphology of maxillary first premolars in the Chinese population. Methods: Eighty recently extracted human maxillary first premolars from a native Chinese population were used. The number and shape of apical constrictions were recorded under a dental operating microscope (DOM) at 12.5X2.5 magnification. After access preparation, a new K-file was inserted into the canal until the tip of the file was just seen at the apical constriction under the DOM. The teeth with files in the canals were X-rayed from a mesiodistal direction using a direct digital radiography (DDR) system, and the distance between the file tip and the center of radiographic apex was directly measured from the computer screen using DDR measurement software. Results: The percentage of teeth with an apical constriction was 78.5% (102/130). The most common apical constriction shapes were oval (55.9%) and round (35.3%). The mean distance between the apical constriction and the anatomical tip of the root was 0.61 mm, and 84.3% (86/102) were within 1 mm. Conclusion: The most common shape of an apical constriction was oval or round, and the distance to the apex was mostly within 1 mm, indicating that root canal therapy should stop 1 mm from the radiographic apex.

  14. Potassium transport across rat alveolar epithelium: evidence for an apical Na+-K+ pump.

    Science.gov (United States)

    Basset, G; Bouchonnet, F; Crone, C; Saumon, G

    1988-06-01

    1. Experiments were performed on rat lungs into which various solutions were instilled whilst the lungs were perfused with either whole blood or Ringer solution. Instillation of ion-free glucose solution led to a net flux of fluid and ions into the alveolar spaces. K+ ions entered faster than Na+ ions and reached a concentration about twice that in the perfusate. Ouabain in the perfusate (basolateral side) prevented the rise in alveolar K+ concentration above that in the perfusate, indicating a transcellular pathway. Ba2+ in the instillate (apical side) hindered the entry of K+ into alveoli, suggesting the presence of apical K+ channels. 2. When Ringer solution was instilled, K+ was continuously removed from the alveoli and the K+ concentration in the instillate remained constant or decreased slightly depending on the rate of fluid absorption. The net K+ efflux from alveoli to blood was 0.23 pmol/(cm2 s). When Ba2+ was added to the instillate the net K+ efflux increased to 0.36 pmol/(cm2 s). Apical ouabain reversed the K+ flux resulting in a net K+ flux of 0.19 pmol/(cm2 s) into the alveoli. This suggests the presence of an Na+-K+-ATPase located in the apical membrane of some alveolar cells. 3. The K+ transport from instillate (Ringer solution) to perfusate was traced by means of 86Rb which was added to the instillate. Ouabain in the instillate did not affect fluid absorption but reduced the apparent 86Rb permeability by 50% although the paracellular permeability (estimated with [3H]mannitol) was unaffected. This also indicates the presence of an apical Na+-K+-ATPase. When ouabain was added to the perfusate, the apparent 86Rb permeability doubled. These findings indicate that recirculation of 86Rb (and K+) occurs due to the activity of both apical and basolateral Na+-K+-ATPases. 4. When ouabain was placed on both sides of the epithelium, preventing transcellular transport, the passive 86Rb permeability was 10.3 x 10(-8) cm/s (assuming an alveolar surface area of

  15. Caveolin-2 Is Required for Apical Lipid Trafficking and Suppresses Basolateral Recycling Defects in the Intestine of Caenorhabditis elegans

    OpenAIRE

    Parker, Scott; Walker, Denise S.; Ly, Sung; Baylis, Howard A.

    2009-01-01

    Caveolins are plasma membrane–associated proteins that colocalize with, and stabilize caveolae. Their functions remain unclear although they are known to be involved in specific events in cell signaling and endocytosis. Caenorhabditis elegans encodes two caveolin genes, cav-1 and cav-2. We show that cav-2 is expressed in the intestine where it is localized to the apical membrane and in intracellular bodies. Using the styryl dye FM4-64 and BODIPY-labeled lactosylceramide, we show that the inte...

  16. Myosins 1 and 6, myosin light chain kinase, actin and microtubules cooperate during antibody-mediated internalisation and trafficking of membrane-expressed viral antigens in feline infectious peritonitis virus infected monocytes.

    Science.gov (United States)

    Dewerchin, Hannah L; Desmarets, Lowiese M; Noppe, Ytse; Nauwynck, Hans J

    2014-02-12

    Monocytes infected with feline infectious peritonitis virus, a coronavirus, express viral proteins in their plasma membranes. Upon binding of antibodies, these proteins are quickly internalised through a new clathrin- and caveolae-independent internalisation pathway. By doing so, the infected monocytes can escape antibody-dependent cell lysis. In the present study, we investigated which kinases and cytoskeletal proteins are of importance during internalisation and subsequent intracellular transport. The experiments showed that myosin light chain kinase (MLCK) and myosin 1 are crucial for the initiation of the internalisation. With co-localisation stainings, it was found that MLCK and myosin 1 co-localise with antigens even before internalisation started. Myosin 6 co-localised with the internalising complexes during passage through the cortical actin, were it might play a role in moving or disintegrating actin filaments, to overcome the actin barrier. One minute after internalisation started, vesicles had passed the cortical actin, co-localised with microtubules and association with myosin 6 was lost. The vesicles were further transported over the microtubules and accumulated at the microtubule organising centre after 10 to 30 min. Intracellular trafficking over microtubules was mediated by MLCK, myosin 1 and a small actin tail. Since inhibiting MLCK with ML-7 was so efficient in blocking the internalisation pathway, this target can be used for the development of a new treatment for FIPV.

  17. An anti-phospholipase A2 receptor quantitative immunoassay and epitope analysis in membranous nephropathy reveals different antigenic domains of the receptor.

    Science.gov (United States)

    Behnert, Astrid; Fritzler, Marvin J; Teng, Beina; Zhang, Meifeng; Bollig, Frank; Haller, Hermann; Skoberne, Andrej; Mahler, Michael; Schiffer, Mario

    2013-01-01

    The phospholipase A2 receptor (PLA2R) was recently discovered as a target autoantigen in patients with idiopathic membranous nephropathy (IMN). Published evidence suggests that the autoantibodies directed towards a conformation dependent epitope are currently effectively detected by a cell based assay (CBA) utilizing indirect immunofluorescence (IIF) on tissue culture cells transfected with the PLA2R cDNA. Limitations of such IIF-CBA assays include observer dependent subjective evaluation of semi-quantitative test results and the protocols are not amenable to high throughput diagnostic testing. We developed a quantitative, observer independent, high throughput capture immunoassay for detecting PLA2R autoantibodies on an addressable laser bead immunoassay (ALBIA) platform. Since reactive domains of PLA2R (i.e. epitopes) could be used to improve diagnostic tests by using small peptides in various high throughput diagnostic platforms, we identified PLA2R epitopes that bound autoantibodies of IMN patients. These studies confirmed that inter-molecular epitope spreading occurs in IMN but use of the cognate synthetic peptides in immunoassays was unable to conclusively distinguish between IMN patients and normal controls. However, combinations of these peptides were able to effectively absorb anti-PLA2R reactivity in IIF-CBA and an immunoassay that employed a lysate derived from HEK cells tranfected with and overexpressing PLA2R. While we provide evidence of intermolecular epitope spreading, our data indicates that in addition to conformational epitopes, human anti-PLA2R reactivity in a commercially available CBA and an addressable laser bead immunoassay is significantly absorbed by peptides representing epitopes of PLA2R.

  18. An anti-phospholipase A2 receptor quantitative immunoassay and epitope analysis in membranous nephropathy reveals different antigenic domains of the receptor.

    Directory of Open Access Journals (Sweden)

    Astrid Behnert

    Full Text Available The phospholipase A2 receptor (PLA2R was recently discovered as a target autoantigen in patients with idiopathic membranous nephropathy (IMN. Published evidence suggests that the autoantibodies directed towards a conformation dependent epitope are currently effectively detected by a cell based assay (CBA utilizing indirect immunofluorescence (IIF on tissue culture cells transfected with the PLA2R cDNA. Limitations of such IIF-CBA assays include observer dependent subjective evaluation of semi-quantitative test results and the protocols are not amenable to high throughput diagnostic testing. We developed a quantitative, observer independent, high throughput capture immunoassay for detecting PLA2R autoantibodies on an addressable laser bead immunoassay (ALBIA platform. Since reactive domains of PLA2R (i.e. epitopes could be used to improve diagnostic tests by using small peptides in various high throughput diagnostic platforms, we identified PLA2R epitopes that bound autoantibodies of IMN patients. These studies confirmed that inter-molecular epitope spreading occurs in IMN but use of the cognate synthetic peptides in immunoassays was unable to conclusively distinguish between IMN patients and normal controls. However, combinations of these peptides were able to effectively absorb anti-PLA2R reactivity in IIF-CBA and an immunoassay that employed a lysate derived from HEK cells tranfected with and overexpressing PLA2R. While we provide evidence of intermolecular epitope spreading, our data indicates that in addition to conformational epitopes, human anti-PLA2R reactivity in a commercially available CBA and an addressable laser bead immunoassay is significantly absorbed by peptides representing epitopes of PLA2R.

  19. The phospholipid flippase ATP8B1 mediates apical localization of the cystic fibrosis transmembrane regulator.

    Science.gov (United States)

    van der Mark, Vincent A; de Jonge, Hugo R; Chang, Jung-Chin; Ho-Mok, Kam S; Duijst, Suzanne; Vidović, Dragana; Carlon, Marianne S; Oude Elferink, Ronald P J; Paulusma, Coen C

    2016-09-01

    Progressive familial intrahepatic cholestasis type 1 (PFIC1) is caused by mutations in the gene encoding the phospholipid flippase ATP8B1. Apart from severe cholestatic liver disease, many PFIC1 patients develop extrahepatic symptoms characteristic of cystic fibrosis (CF), such as pulmonary infection, sweat gland dysfunction and failure to thrive. CF is caused by mutations in the cystic fibrosis transmembrane conductance regulator (CFTR), a chloride channel essential for epithelial fluid transport. Previously it was shown that CFTR transcript levels were strongly reduced in livers of PFIC1 patients. Here we have investigated the hypothesis that ATP8B1 is important for proper CFTR expression and function. We analyzed CFTR expression in ATP8B1-depleted intestinal and pulmonary epithelial cell lines and assessed CFTR function by measuring short-circuit currents across transwell-grown ATP8B1-depleted intestinal T84 cells and by a genetically-encoded fluorescent chloride sensor. In addition, we studied CFTR surface expression upon induction of CFTR transcription. We show that CFTR protein levels are strongly reduced in the apical membrane of human ATP8B1-depleted intestinal and pulmonary epithelial cell lines, a phenotype that coincided with reduced CFTR activity. Apical membrane insertion upon induction of ectopically-expressed CFTR was strongly impaired in ATP8B1-depleted cells. We conclude that ATP8B1 is essential for correct apical localization of CFTR in human intestinal and pulmonary epithelial cells, and that impaired CFTR localization underlies some of the extrahepatic phenotypes observed in ATP8B1 deficiency. PMID:27301931

  20. Effect of cytokinins on shoot apical meristem in Nicotiana tabacum

    Directory of Open Access Journals (Sweden)

    Uzelac Branka

    2012-01-01

    Full Text Available Cytokinins are involved in plant cell proliferation leading to plant growth and morphogenesis. The size, activity and maintenance of the shoot apical meristem (SAM are defined by a balanced rate of mitotic cell divisions and functional cell differentiation that are controlled by cytokinins. In order to investigate the effect of exogenous cytokinin on SAM, morpho-anatomical changes in the shoot apices of tobacco treated with benzyladenine (BA were compared to those of untreated control plants.

  1. Effect of cytokinins on shoot apical meristem in Nicotiana tabacum

    OpenAIRE

    Uzelac Branka; Janošević Dušica; Stojičić Dragana; Budimir Snežana

    2012-01-01

    Cytokinins are involved in plant cell proliferation leading to plant growth and morphogenesis. The size, activity and maintenance of the shoot apical meristem (SAM) are defined by a balanced rate of mitotic cell divisions and functional cell differentiation that are controlled by cytokinins. In order to investigate the effect of exogenous cytokinin on SAM, morpho-anatomical changes in the shoot apices of tobacco treated with benzyladenine (BA) were compared to those of untreated control...

  2. Pseudoaneurysm of the left ventricle following apical approach TAVI

    OpenAIRE

    Vanezis Andrew P; Baig Mirza K; Mitchel Ian M; Shajar Matloob; Naik Surendra K; Henderson Robert A; Mathew Thomas

    2011-01-01

    Abstract Symptomatic severe aortic stenosis carries a two year survival of only 50%. However many patients are unsuitable for conventional aortic valve replacement as they are considered too high risk due to significant co-morbidities. Transcatheter Aortic Valve Implantation (TAVI) offers a viable alternative for this high risk patient group, either by the femoral or apical route. This article reports a case of a pseudoaneurysm of the left ventricle following an apical approach TAVI in an eld...

  3. Haemostatic agents in apical surgery. A systematic review

    Science.gov (United States)

    Clé-Ovejero, Adrià

    2016-01-01

    Background Blood presence in apical surgery can prevent the correct vision of the surgical field, change the physical properties of filling materials and reduce their sealing ability. Objetive To describe which are the most effective and safest haemostatic agents to control bleeding in patients undergoing apical surgery. Material and Methods TWe carried out a systematic review, using Medline and Cochrane Library databases, of human clinical studies published in the last 10 years. Results The agents that proved more effective in bleeding control were calcium sulphate (100%) and collagen plus epinephrine (92.9%) followed by ferric sulphate (60%), gauze packing (30%) and collagen (16.7%). When using aluminium chloride (Expasyl®), over 90% of the apical lesions improved, but this agent seemed to increase swelling. Epinephrine with collagen did not significantly raise either blood pressure or heart rate. Conclusions Despite the use of several haemostatic materials in apical surgery, there is little evidence on their effectiveness and safety. The most effective haemostatic agents were calcium sulphate and epinephrine plus collagen. Epinephrine plus collagen did not seem to significantly raise blood pressure or heart rate during surgery. Aluminium chloride did not increase postoperative pain but could slightly increase postoperative swelling. Randomized clinical trials are needed to assess the haemostatic effectiveness and adverse effects of haemostatic materials in apical surgery. Key words:Haemostasis, apical surgery. PMID:27475689

  4. Shot and Patronin polarise microtubules to direct membrane traffic and biogenesis of microvilli in epithelia.

    Science.gov (United States)

    Khanal, Ichha; Elbediwy, Ahmed; Diaz de la Loza, Maria Del Carmen; Fletcher, Georgina C; Thompson, Barry J

    2016-07-01

    In epithelial tissues, polarisation of microtubules and actin microvilli occurs along the apical-basal axis of each cell, yet how these cytoskeletal polarisation events are coordinated remains unclear. Here, we examine the hierarchy of events during cytoskeletal polarisation in Drosophila melanogaster epithelia. Core apical-basal polarity determinants polarise the spectrin cytoskeleton to recruit the microtubule-binding proteins Patronin (CAMSAP1, CAMSAP2 and CAMPSAP3 in humans) and Shortstop [Shot; MACF1 and BPAG1 (also known as DST) in humans] to the apical membrane domain. Patronin and Shot then act to polarise microtubules along the apical-basal axis to enable apical transport of Rab11 endosomes by the Nuf-Dynein microtubule motor complex. Finally, Rab11 endosomes are transferred to the MyoV (also known as Didum in Drosophila) actin motor to deliver the key microvillar determinant Cadherin 99C to the apical membrane to organise the biogenesis of actin microvilli. PMID:27231092

  5. Effect of apical clearing technique on the treatment outcome of teeth with asymptomatic apical periodontitis: A randomized clinical trial

    Science.gov (United States)

    Mittal, Priya; Logani, Ajay; Shah, Naseem; Pandey, R. M.

    2016-01-01

    Aim: This study aims to compare the periapical healing of teeth with asymptomatic apical periodontitis treated either by conventional apical preparation (CAP) or apical clearing technique (ACT). Materials and Methods: Twenty subjects with bilateral nonvital similar teeth exhibiting comparable periapical index (PAI) score were enrolled and randomly allocated. Group I (CAP, n = 20): Apical preparation three sizes greater (master apical file [MAF]) than the first binding file at the established working length. Group II (ACT, n = 20): Apical preparation three sizes greater than the MAF that was followed by dry reaming. Root canal therapy was accomplished in single-visit for all the teeth. They were pursued radiographically at 3, 6, 9 and 12 months. Pre- and post-treatment PAI scores were compared. To ascertain the proportion of healed teeth between the two groups, McNemar Chi-square test was applied. Results: At 3, 6, and 9 months’ time interval the proportion of healed teeth for Group II (ACT) was greater in comparison to Group I (CAP) (P < 0.05). However, at 12 months follow-up period this difference was not significant (P = 0.08). Conclusion: ACT enhanced the healing kinetics. However, the long-term (12 months) radiographic outcome was similar for either technique. PMID:27656054

  6. Differentiation of Apical Bud Cells in a Newly Developed Apical Bud Transplantation Model Using GFP Transgenic Mice as Donor

    Science.gov (United States)

    Sakagami, Ryuji; Yoshinaga, Yasunori; Okamura, Kazuhiko

    2016-01-01

    Rodent mandibular incisors have a unique anatomical structure that allows teeth to grow throughout the lifetime of the rodent. This report presents a novel transplantation technique for studying the apical bud differentiation of rodent mandibular incisors. Incisal apical end tissue with green fluorescent protein from transgenic mouse was transplanted to wild type mice, and the development of the transplanted cells were immunohistologically observed for 12 weeks after the transplantation. Results indicate that the green fluorescent apical end tissue replaced the original tissue, and cells from the apical bud differentiated and extended toward the incisal edge direction. The immunostaining with podoplanin also showed that the characteristics of the green fluorescent tissue were identical to those of the original. The green fluorescent cells were only found in the labial side of the incisor up to 4 weeks. After 12 weeks, however, they were also found in the lingual side. Here the green fluorescent cementocyte-like cells were only present in the cementum close to the dentin surface. This study suggests that some of the cells that form the cellular cementum come from the apical tissue including the apical bud in rodent incisors. PMID:26978064

  7. Changes of the Unique Odontogenic Properties of Rat Apical Bud Cells under the Developing Apical Complex Microenvironment

    Institute of Scientific and Technical Information of China (English)

    Jun Fang; Liang Tang; Xiao-hui Liu; Ling-ying Wen; Yan Jin

    2009-01-01

    Aim To characterize the odontogenic capability of apical bud and phenotypical change of apical bud cells (ABCs) in different microenvironment. Methodology Incisor apical bud tissues from neonatal SD rat were dissected and transplanted into the renal capsules to determine their odontogenic capability. Meanwhile ABCs were cultured and purified by repeated differential trypsinization. Then ABCs were cultured with conditioned medium from developing apical complex cells (DAC-CM). Immunocytochemistry, reverse transcriptase polymerase chain reaction (RT-PCR) and scanning electron microscope (SEM) were performed to compare the biological change of ABC treated with or without DAC-CM. Results First we confirmed the ability of apical bud to form crown-like structure ectopically. Equally important, by using the developing apical complex (DAC) conditioned medium, we found the microenvironment created by root could abrogate the "crown" features of ABCs and promote their proliferation and differentiation. Conclusion ABCs possess odontogenic capability to form crown-like tissues and this property can be affected by root-produced microenvironment.

  8. Five-year longitudinal assessment of the prognosis of apical microsurgery

    DEFF Research Database (Denmark)

    von Arx, Thomas; Jensen, Simon S; Hänni, Stefan;

    2012-01-01

    Apical surgery is an important treatment option for teeth with post-treatment apical periodontitis. Knowledge of the long-term prognosis is necessary when weighing apical surgery against alternative treatments. This study assessed the 5-year outcome of apical surgery and its predictors in a cohor...

  9. Expression of leukemic stem cell associated membrane antigens in acute leukemia cells%白血病干细胞相关抗原在急性白血病细胞中的表达

    Institute of Scientific and Technical Information of China (English)

    肖平; 曾耀英; 林蔚

    2011-01-01

    目的 探讨白血病干细胞(leukemia stem cell,LSC)相关抗原在不同亚型急性白血病细胞中的表达规律.方法 采用流式细胞术检测LSC相关抗原CD96,CD90,CD123,CD71等在50例不同亚型急性白血病细胞中的表达,包括急性粒细胞白血病未分化型(M1)、急性粒细胞白血病部分分化型(M2)、急性早幼粒细胞白血病(Ms)、急性粒-单核细胞白血病(M4)和急性B淋巴细胞白血病.结果 CD96在M.的表达率(90.00%)明显高于M2(18.18%)和急性B淋巴细胞白血病(20.00%)(P<0.05);各亚型急性白血病均表达CD123,但差异无统计学意义(P>0.05) ;CD71在急性髓细胞白血病各亚型中(M1、M2、M3和M4)阳性表达率分别为80.00%、72.73%、90.00%和100.00%,明显高于急性B淋巴细胞白血病(P<0.05);CD90在急性B淋巴细胞白血病中阳性表达率为13.33%,高于急性髓细胞白血病(P>0.05).结论 CD71与CD96的表达有亚型特异性,CD96可能具有指示系列分化和细胞分化程度的作用,CD71可用于区分急性淋巴细胞白血病和急性髓细胞白血病.%Objective To study the expression of leukemic stem cell associated membrane antigens in acute leukemia cells with different subtypes. Methods Leukemic stem cell associated membrane antigens CD96, CD90, CD123 and CD71 were detected with flow cytometry in 50 acute leukemia patients with different subtypes including acute myeloblast leukemia without cell maturation (M1), acute myeloblast leukemia with cell maturation (M2), acute promyelocytic leukemia (M3), acute myelomonocytic leukemia (AMML) and B-acute lymphoblastic leukemia (B-ALL). Results The positive rate of CD96 expression in M3(90. 00%) was significantly higher than that in M2(18. 18%) and B-ALL (20. 00%)(P 0. 05). The positive rate of CD71 expression in four subtypes of acute myeloid leukemia (M1 , M2, M3 and M4) was 80. 00%, 72. 73%, 90. 00% and 100. 00% respectively, all of which were significantly higher than

  10. Impact of Inhibiting Ileal Apical Versus Basolateral Bile acid Transport on Cholesterol Metabolism and Atherosclerosis in Mice

    Science.gov (United States)

    Dawson, Paul A.

    2015-01-01

    Background Bile acid sequestrants have been used for many years to treat hypercholesterolemia by increasing hepatic conversion of cholesterol to bile acids, thereby inducing hepatic LDL receptor expression and clearance of apoB-containing particles. In order to further understand the underlying molecular mechanisms linking gut-liver signaling and cholesterol homeostasis, mouse models defective in ileal apical membrane bile acid transport (Asbt null) and ileal basolateral membrane bile acid transport (Ostα null) were studied under basal and hypercholesterolemic conditions. Key Messages Hepatic conversion of cholesterol to bile acids is the major pathway for cholesterol catabolism and a major mechanism for cholesterol elimination. Blocking ileal apical membrane bile acid transport (Asbt null mice) increases fecal bile acid excretion, hepatic Cyp7a1 expression and the relative proportion of taurocholate in the bile acid pool, but decreases ileal FGF15 expression, bile acid pool size, and hepatic cholesterol content. In contrast, blocking ileal basolateral membrane bile acid transport (Ostα null mice) increases ileal FGF15 expression, reduces hepatic Cyp7a1 expression, and increases the proportion of tauro-β-muricholic acid in the bile acid pool. In the hypercholesterolemic apoE null background, plasma cholesterol levels and measurements of atherosclerosis were reduced in Asbt/apoE null mice but not in Ostα/apoE null mice. Conclusions Blocking intestinal absorption of bile acids at the apical versus basolateral membrane differentially affects bile acid and cholesterol metabolism, including the development of hypercholesterolemia-associated atherosclerosis. The molecular mechanism likely involves altered regulation of ileal FGF15 expression. PMID:26045273

  11. Cross-reactivity of anti-PLA2R1 autoantibodies to rabbit and mouse PLA2R1 antigens and development of two novel ELISAs with different diagnostic performances in idiopathic membranous nephropathy.

    Science.gov (United States)

    Seitz-Polski, Barbara; Dolla, Guillaume; Payré, Christine; Tomas, Nicola M; Lochouarn, Marine; Jeammet, Louise; Mariat, Christophe; Krummel, Thierry; Burtey, Stéphane; Courivaud, Cécile; Schlumberger, Wolfgang; Zorzi, Kévin; Benzaken, Sylvia; Bernard, Ghislaine; Esnault, Vincent L M; Lambeau, Gérard

    2015-11-01

    About 70% of patients with idiopathic membranous nephropathy (iMN) have autoantibodies to the phospholipase A2 receptor PLA2R1. We screened sera from iMN patients for their cross-reactivity to human (h), rabbit (rb) and mouse (m) PLA2R1 by western blot (WB) and antigen-specific ELISAs. All iMN patients recognized hPLA2R1 and rbPLA2R1 by WB, and a rbPLA2R1 ELISA was as sensitive as the standardized hPLA2R1 ELISA to monitor anti-PLA2R1 in patients with active disease or in drug-induced remission. In contrast, only 51% of patients were reactive to mPLA2R1 by WB, and a maximum of 78% were weakly to highly positive in the mPLA2R1 ELISA, suggesting that iMN patients exhibit different subsets of anti-PLA2R1 autoantibodies against epitopes that are shared or not among PLA2R1 orthologs. In a cohort of 41 patients with a mean follow-up of 42 months from anti-PLA2R1 assay, the detection of anti-mPLA2R1 autoantibodies was an independent predictor of clinical outcome in multivariate analysis (p = 0.009), and a ROC curve analysis identified a threshold of 605 RU/mL above which 100% of patients (12 patients) had a poor renal outcome (p 605 RU/mL) only in iMN patients at risk of poor prognosis, and is thus useful to predict iMN outcome. PMID:26296473

  12. Deletion of the anion exchanger Slc26a4 (pendrin) decreases apical Cl−/HCO3− exchanger activity and impairs bicarbonate secretion in kidney collecting duct

    OpenAIRE

    Amlal, Hassane; Petrovic, Snezana; Xu, Jie; Wang, Zhaohui; Sun, Xuming; Barone, Sharon; Soleimani, Manoocher

    2010-01-01

    The anion exchanger Pendrin, which is encoded by SLC26A4 (human)/Slc26a4 (mouse) gene, is localized on the apical membrane of non-acid-secreting intercalated (IC) cells in the kidney cortical collecting duct (CCD). To examine its role in the mediation of bicarbonate secretion in vivo and the apical Cl−/HCO3− exchanger in the kidney CCD, mice with genetic deletion of pendrin were generated. The mutant mice show the complete absence of pendrin expression in their kidneys as assessed by Northern...

  13. Bony change of apical lesion healing process using fractal analysis

    International Nuclear Information System (INIS)

    To investigate the change of bone healing process after endodontic treatment of the tooth with an apical lesion by fractal analysis. Radiographic images of 35 teeth from 33 patients taken on first diagnosis, 6 months, and 1 year after endodontic treatment were selected. Radiographic images were taken by JUPITER computerized Dental X-ray System. Fractal dimensions were calculated three times at each area by Scion Image PC program. Rectangular region of interest (30 x 30) were selected at apical lesion and normal apex of each image. The fractal dimension at apical lesion of first diagnosis (L0) is 0.940 ± 0.361 and that of normal area (N0) is 1.186 ± 0.727 (p1) is 1.076 ± 0.069 and that of normal area (N1) is 1.192 ± 0.055 (p2) is 1.163 ± 0.074 and that of normal area (N2) is 1.225 ± 0.079 (p<0.05). After endodontic treatment, the fractal dimensions at each apical lesions depending on time showed statistically significant difference. And there are statistically significant different between normal area and apical lesion on first diagnosis, 6 months after, 1 year after. But the differences were grow smaller as time flows. The evaluation of the prognosis after the endodontic treatment of the apical lesion was estimated by bone regeneration in apical region. Fractal analysis was attempted to overcome the limit of subjective reading, and as a result the change of the bone during the healing process was able to be detected objectively and quantitatively.

  14. Apical secretion of apolipoproteins from enterocytes

    DEFF Research Database (Denmark)

    Danielsen, E M; Hansen, Gert Helge; Poulsen, Mona Dam

    1993-01-01

    Synthesis and secretion of apolipoproteins in pig small intestine was studied by pulse-chase labeling of jejunal segments, kept in organ culture. Apo A-1 and apo B-48 were the two major proteins released, constituting 25 and 10%, respectively, of the total amount of labeled protein in the mucosal...... that enterocytes release most of their newly made free apo A-1 and a significant portion of apo B-48 by exocytosis via the brush border membrane into the intestinal lumen. Fat absorption reduced apolipoprotein secretion to the medium and induced the formation of chylomicrons, containing apo A-1 at their surface......-side medium where they appeared with a t1/2 of 50-60 min. Using tissue from fasting animals, > 85% of newly synthesized apo A-1 and about one third of apo B-48 was released to the mucosal-side medium. Newly synthesized apolipoprotein that remained associated with the intestinal segment accumulated...

  15. Lipid rafts in epithelial brush borders: atypical membrane microdomains with specialized functions

    DEFF Research Database (Denmark)

    Danielsen, E Michael; Hansen, Gert H

    2003-01-01

    of the apical surface sterically accessible for membrane fusion/budding events. Many of these invaginations appear as pleiomorphic, deep apical tubules that extend up to 0.5-1 microm into the underlying terminal web region. Their sensitivity to methyl-beta-cyclodextrin suggests them to contain cholesterol...

  16. Cryopreservation of Pelargonium apices by droplet-vitrification.

    Science.gov (United States)

    Gallard, Anthony; Panis, Bart; Dorion, Nöelle; Swennen, Rony; Grapin, Agnès

    2008-01-01

    The droplet-vitrification method was adapted to Pelargonium apices by optimizing the duration of the loading solution (LS) as well as the plant vitrification solution 2 (PVS2). The excised apices were dehydrated in two steps (20 min in LS and 15 min in PVS2) and then immersed directly in liquid nitrogen (LN). After thawing and unloading in the recovery solution at room temperature for 15 min, apices were plated onto semi-solid Murashige and Skoog medium. This simple protocol without any pretreatment was successfully applied to eight cultivars with a survival level ranging between 55.6 - 96.2 percent and a regrowth level between 9.1 and 70.6 percent. These results prove the feasibility of the long-term storage of Pelargonium germplasm through cryopreservation.

  17. Antigenic Variation in Plasmodium falciparum.

    Science.gov (United States)

    Petter, Michaela; Duffy, Michael F

    2015-01-01

    Plasmodium falciparum is the protozoan parasite that causes most malaria-associated morbidity and mortality in humans with over 500,000 deaths annually. The disease symptoms are associated with repeated cycles of invasion and asexual multiplication inside red blood cells of the parasite. Partial, non-sterile immunity to P. falciparum malaria develops only after repeated infections and continuous exposure. The successful evasion of the human immune system relies on the large repertoire of antigenically diverse parasite proteins displayed on the red blood cell surface and on the merozoite membrane where they are exposed to the human immune system. Expression switching of these polymorphic proteins between asexual parasite generations provides an efficient mechanism to adapt to the changing environment in the host and to maintain chronic infection. This chapter discusses antigenic diversity and variation in the malaria parasite and our current understanding of the molecular mechanisms that direct the expression of these proteins. PMID:26537377

  18. Phosphatidylcholine passes through lateral tight junctions for paracellular transport to the apical side of the polarized intestinal tumor cell-line CaCo2.

    Science.gov (United States)

    Stremmel, Wolfgang; Staffer, Simone; Gan-Schreier, Hongying; Wannhoff, Andreas; Bach, Margund; Gauss, Annika

    2016-09-01

    Phosphatidylcholine (PC) is the most abundant phospholipid in intestinal mucus, indicative of a specific transport system across the mucosal epithelium to the intestinal lumen. To elucidate this transport mechanism, we employed a transwell tissue culture system with polarized CaCo2 cells. It was shown that PC could not substantially be internalized by the cells. However, after basal application of increasing PC concentrations, an apical transport of 47.1±6.3nmolh(-1)mMPC(-1) was observed. Equilibrium distribution studies with PC applied in equal concentrations to the basal and apical compartments showed a 1.5-fold accumulation on the expense of basal PC. Disruption of tight junctions (TJ) by acetaldehyde or PPARγ inhibitors or by treatment with siRNA to TJ proteins suppressed paracellular transport by at least 50%. Transport was specific for the choline containing the phospholipids PC, lysoPC and sphingomyelin. We showed that translocation is driven by an electrochemical gradient generated by apical accumulation of Cl(-) and HCO3(-) through CFTR. Pretreatment with siRNA to mucin 3 which anchors in the apical plasma membrane of mucosal cells inhibited the final step of luminal PC secretion. PC accumulates in intestinal mucus using a paracellular, apically directed transport route across TJs. PMID:27365309

  19. Apical transport of influenza A virus ribonucleoprotein requires Rab11-positive recycling endosome.

    Directory of Open Access Journals (Sweden)

    Fumitaka Momose

    Full Text Available Influenza A virus RNA genome exists as eight-segmented ribonucleoprotein complexes containing viral RNA polymerase and nucleoprotein (vRNPs. Packaging of vRNPs and virus budding take place at the apical plasma membrane (APM. However, little is known about the molecular mechanisms of apical transport of newly synthesized vRNP. Transfection of fluorescent-labeled antibody and subsequent live cell imaging revealed that punctate vRNP signals moved along microtubules rapidly but intermittently in both directions, suggestive of vesicle trafficking. Using a series of Rab family protein, we demonstrated that progeny vRNP localized to recycling endosome (RE in an active/GTP-bound Rab11-dependent manner. The vRNP interacted with Rab11 through viral RNA polymerase. The localization of vRNP to RE and subsequent accumulation to the APM were impaired by overexpression of Rab binding domains (RBD of Rab11 family interacting proteins (Rab11-FIPs. Similarly, no APM accumulation was observed by overexpression of class II Rab11-FIP mutants lacking RBD. These results suggest that the progeny vRNP makes use of Rab11-dependent RE machinery for APM trafficking.

  20. AmotL2 disrupts apical-basal cell polarity and promotes tumour invasion.

    Science.gov (United States)

    Mojallal, Mahdi; Zheng, Yujuan; Hultin, Sara; Audebert, Stéphane; van Harn, Tanja; Johnsson, Per; Lenander, Claes; Fritz, Nicolas; Mieth, Christin; Corcoran, Martin; Lembo, Frédérique; Hallström, Marja; Hartman, Johan; Mazure, Nathalie M; Weide, Thomas; Grandér, Dan; Borg, Jean-Paul; Uhlén, Per; Holmgren, Lars

    2014-01-01

    The establishment and maintenance of apical-basal cell polarity is essential for the functionality of glandular epithelia. Cell polarity is often lost in advanced tumours correlating with acquisition of invasive and malignant properties. Despite extensive knowledge regarding the formation and maintenance of polarity, the mechanisms that deregulate polarity in metastasizing cells remain to be fully characterized. Here we show that AmotL2 expression correlates with loss of tissue architecture in tumours from human breast and colon cancer patients. We further show that hypoxic stress results in activation of c-Fos-dependent expression of AmotL2 leading to loss of polarity. c-Fos/hypoxia-induced p60 AmotL2 interacts with the Crb3 and Par3 polarity complexes retaining them in large vesicles and preventing them from reaching the apical membrane. The resulting loss of polarity potentiates the response to invasive cues in vitro and in vivo in mice. These data provide a molecular mechanism how hypoxic stress deregulates cell polarity during tumour progression. PMID:25080976

  1. IN VIVO ANALYSIS OF SOME KEY CHARACTERISTICS OF THE APICAL ZONE IN TEETH WITH CHRONIC APICAL PERIODONTITIS.

    Directory of Open Access Journals (Sweden)

    Angela Gusiyska

    2014-11-01

    Full Text Available Introduction: The pathogenesis of internal and external resorptive processes in the dental tissues and those of the periapical zone is not fully understood, but the main purpose, either in teeth with internal resorption or in teeth with periapical lesions, is decontamination of the endodontic space and subsequent three-dimensional obturation in order to isolate periapical and oral tissues and prevent reinfection. Purpose: The aim of this article is to analyze in vivo some key characteristics of the apical zone in teeth with chronic apical periodontitis. Material and Methods: To facilitate the clinical protocol after the radiographic analysis and assessment of patency, the working lengths of 153 root canals (n = 153 in 106 teeth were determined. The clinical widths of the apical narrowing were measured by using the last instrument (ISO 0.02 tapered file, which can move freely through the apical narrowing after electrometric determination of the working length (Raypex 5 /VDW, Germany/. Results and Discussion: Determination of working width and working length is important for realizing the first stage of decontamination – maximum instrumentation of the endodontic space and choice of a clinical protocol. The classification of root canals in a particular group according to the relative patency or lysed apical opening is important for the selection of obturation technique, which is essential for reducing the microleakage in the zone. Conclusion: Since the target of this work were teeth with CAP, in the majority of the cases with clinical findings of root canals with preexisting filling, radicular pins, obliteration, separated canal instruments, perforations at different levels, via falsa or thresholds, the access to the apical zone was not subjected to a closely observed instrumental clinical protocol. In the treatment of each case, however, the clinical principles of modern endodontic treatment were closely observed.

  2. Pre-therapeutic dosimetry of normal organs and tissues of {sup 177}Lu-PSMA-617 prostate-specific membrane antigen (PSMA) inhibitor in patients with castration-resistant prostate cancer

    Energy Technology Data Exchange (ETDEWEB)

    Kabasakal, Levent; AbuQbeitah, Mohammad; Ayguen, Aslan; Yeyin, Nami [Istanbul University, Department of Nuclear Medicine, Cerrahpasa Medical Faculty, Istanbul (Turkey); Ocak, Meltem [Istanbul University, Department of Pharmaceutical Technology, Pharmacy Faculty, Istanbul (Turkey); Demirci, Emre [Sisli Etfal Training and Research Hospital, Department of Nuclear Medicine, Istanbul (Turkey); Toklu, Turkay [Yeditepe University Medical Faculty, Department of Nuclear Medicine, Istanbul (Turkey)

    2015-12-15

    {sup 177}Lu-617-prostate-specific membrane antigen (PSMA) ligand seems to be a promising tracer for radionuclide therapy of progressive prostate cancer. However, there are no published data regarding the radiation dose given to the normal tissues. The aim of the present study was to estimate the pretreatment radiation doses in patients who will undergo radiometabolic therapy using a tracer amount of {sup 177}Lu-labeled PSMA ligand. The study included seven patients with progressive prostate cancer with a mean age of 63.9 ± 3.9 years. All patients had prior PSMA positron emission tomography (PET) imaging and had intense tracer uptake at the lesions. The injected {sup 177}Lu-PSMA-617 activity ranged from 185 to 210 MBq with a mean of 192.6 ± 11.0 MBq. To evaluate bone marrow absorbed dose 2-cc blood samples were withdrawn in short variable times (3, 15, 30, 60, and 180 min and 24, 48, and 120 h) after injection. Whole-body images were obtained at 4, 24, 48, and 120 h post-injection (p.i.). The geometric mean of anterior and posterior counts was determined through region of interest (ROI) analysis. Attenuation correction was applied using PSMA PET/CT images. The OLINDA/EXM dosimetry program was used for curve fitting, residence time calculation, and absorbed dose calculations. The calculated radiation-absorbed doses for each organ showed substantial variation. The highest radiation estimated doses were calculated for parotid glands and kidneys. Calculated radiation-absorbed doses per megabecquerel were 1.17 ± 0.31 mGy for parotid glands and 0.88 ± 0.40 mGy for kidneys. The radiation dose given to the bone marrow was significantly lower than those of kidney and parotid glands (p < 0.05). The calculated radiation dose to bone marrow was 0.03 ± 0.01 mGy/MBq. Our first results suggested that {sup 177}Lu-PSMA-617 therapy seems to be a safe method. The dose-limiting organ seems to be the parotid glands rather than kidneys and bone marrow. The lesion radiation doses are

  3. Distinct roles of Bazooka and Stardust in the specification of Drosophila photoreceptor membrane architecture.

    Science.gov (United States)

    Hong, Yang; Ackerman, Larry; Jan, Lily Yeh; Jan, Yuh-Nung

    2003-10-28

    Photoreceptors form during Drosophila pupal development and acquire elaborate membrane structures, including the rhabdomeres and stalk membranes. Here, we show that the development of these cellular structures involves two distinct processes: the establishment of apical-basal polarity that requires Bazooka (Baz), and the regionalization of apical membrane into stalk membranes and rhabdomeres that requires Stardust (Sdt). In the absence of Baz, the apical-basal polarity is compromised in early pupal photoreceptors, and no identifiable apical membrane domain is formed. Sdt, in contrast, plays a more limited role in apical-basal polarity but is essential for the proper localization of transmembrane protein Crumbs (Crb), known to be required in the biogenesis of stalk membrane. Loss of Sdt causes strong defects in stalk membrane and rhabdomere resembling crb mutant phenotype. Thus, proteins required for establishing the early embryonic epithelial polarity are used later for the morphogenesis of photoreceptors, with Baz and Sdt functioning in different aspects of the formation of the apical-basal cellular architecture.

  4. [Nonsurgical retreatment in a case of a radiolucent apical lesion].

    Science.gov (United States)

    Vicente Gómez, A; Rodríguez Ponce, A

    1989-01-01

    We present a case of failure that was helpful solved without surgical endodontic treatment. We don't achieve clinical success besides endodontic treatment was twice remade. Finally we decided to put a temporary filling with calcium hydroxide and wait until apical radiolucency disappear and complete our treatment with gutta-percha, sealer and lateral condensation. PMID:2640036

  5. Assessment of apical periodontitis by MRI. A feasibility study

    Energy Technology Data Exchange (ETDEWEB)

    Geibel, M.A. [Ulm Univ. (Germany). Oral and Maxillofacial Surgery; Schreiber, E.S.; Bracher, A.K.; Rasche, V. [Ulm Univ. (Germany). Internal Medicine II; Hell, E.; Ulrici, J. [Sirona Dental Systems GmbH, Bensheim (Germany). Dental Imaging; Sailer, L.K. [DOC Praxisklinik im Wiley, Neu-Ulm (Germany). MKG; Ozpeynirci, Y. [Ulm Univ. (Germany). Diagnostic and Interventional Radiology

    2015-04-15

    The purpose of this clinical feasibility study was to evaluate the applicability of magnetic resonance imaging (MRI) for the assessment of apical periodontitis in direct comparison with cone beam CT (CBCT). 19 consecutive patients (average age 43 ± 13 years) with 34 lesions in total (13 molars, 14 premolars and 7 front teeth) were enrolled in this feasibility study. Periapical lesions were defined as periapical radiolucencies (CBCT) or structural changes in the spongy bone signal (MRI), which were connected with the apical part of a root and with at least twice the width of the periodontal ligament space. The location and dimension of the lesions were compared between MRI and CBCT. While mainly mineralized tissue components such as teeth and bone were visible with CBCT, complimentary information of the soft tissue components was assessable with MRI. The MRI images provided sufficient diagnostic detail for the assessment of the main structures of interest. Heterogeneous contrast was observed within the lesion, with often a clear enhancement close to the apical foramen and the periodontal gap. No difference for lesion visibility was observed between MRI and CBCT. The lesion dimensions corresponded well, but were slightly but significantly overestimated with MRI. A heterogeneous lesion appearance was observed in several patients. Four patients presented with a well circumscribed hyperintense signal in the vicinity of the apical foramen. The MRI capability of soft tissue characterization may facilitate detailed analysis of periapical lesions. This clinical study confirms the applicability of multi-contrast MRI for the identification of periapical lesions.

  6. [Nonsurgical retreatment in a case of a radiolucent apical lesion].

    Science.gov (United States)

    Vicente Gómez, A; Rodríguez Ponce, A

    1989-01-01

    We present a case of failure that was helpful solved without surgical endodontic treatment. We don't achieve clinical success besides endodontic treatment was twice remade. Finally we decided to put a temporary filling with calcium hydroxide and wait until apical radiolucency disappear and complete our treatment with gutta-percha, sealer and lateral condensation.

  7. Hypertrophic cardiomyopathy with apical left ventricular aneurysm: a case report

    Institute of Scientific and Technical Information of China (English)

    江腾勇; 韩智红; 王京; 吕强; 吴学思

    2002-01-01

    @@ Morphological diversity is one characteristic of hypertrophic cardi omyopathy (HCM), but it is not common that HCM is associated with apical left ve ntricular aneurysm (LVA) without evidence of a coronary artery lesion. We repor t on such a case and review the pathogenesis, manifestations and diagnostic meth ods by collecting the few available papers published on this topic.

  8. Histocompatibility antigen test

    Science.gov (United States)

    ... more common in certain autoimmune diseases . For example, HLA-B27 antigen is found in many people (but not ... More Ankylosing spondylitis Autoimmune disorders Bone marrow transplant HLA-B27 antigen Kidney transplant Reactive arthritis Update Date 2/ ...

  9. Protecting capacity against malaria of chemically defined tetramer forms based on the Plasmodium falciparum apical sushi protein as potential vaccine components.

    Science.gov (United States)

    Vanegas, Magnolia; Bermúdez, Adriana; Guerrero, Yuly Andrea; Cortes-Vecino, Jesús Alfredo; Curtidor, Hernando; Patarroyo, Manuel Elkin; Lozano, José Manuel

    2014-08-15

    Developing novel generations of subunit-based antimalarial vaccines in the form of chemically-defined macromolecule systems for multiple antigen presentation represents a classical problem in the field of vaccine development. Many efforts involving synthesis strategies leading to macromolecule constructs have been based on dendrimer-like systems, the condensation of large building blocks and conventional asymmetric double dimer constructs, all based on lysine cores. This work describes novel symmetric double dimer and condensed linear constructs for presenting selected peptide multi-copies from the apical sushi protein expressed in Plasmodium falciparum. These molecules have been proved to be safe and innocuous, highly antigenic and have shown strong protective efficacy in rodents challenged with two Plasmodium species. Insights into systematic design, synthesis and characterisation have led to such novel antigen systems being used as potential platforms for developing new anti-malarial vaccine candidates. PMID:25063026

  10. Multilabel immunofluorescence and antigen reprobing on formalin-fixed paraffin-embedded sections: novel applications for precision pathology diagnosis.

    Science.gov (United States)

    Pan, Jie; Thoeni, Cornelia; Muise, Aleixo; Yeger, Herman; Cutz, Ernest

    2016-06-01

    We report new methods for multilabel immunofluorescence (MIF) and reprobing of antigen epitopes on the same formalin-fixed paraffin-embedded (FFPE) sections. The MIF method includes an antigen-retrieval step followed by multilabel immunostaining and examination by confocal microscopy. As examples, we illustrate epitopes localized to the apical and basolateral membranes, and the cytoplasm of enterocytes of normal small intestine and in cases of congenital enteropathies (microvillous inclusion disease and congenital tufting enteropathy). We also demonstrate localization of the bile salt excretion pump protein (BSEP) in bile canalicular membrane of normal hepatocytes and in cases of primary sclerosing cholangitis. To demonstrate colocalization of cytoplasmic and nuclear epitopes we analyzed normal control and hyperplastic pulmonary neuroendocrine cells (PNEC) and neuroepithelial bodies (NEBs), presumed airway sensors in the lungs of infants with bronchopulmonary dysplasia (BPD). As cytoplasmic markers we used anti-bombesin or anti-synaptic vesicle protein 2 (SV2) antibody, respectively, and for nuclear localization, antibodies against neurogenic genes mammalian achaete-scute homolog (Mash1) and prospero homeobox 1 (Prox1), essential for NEB cells differentiation and maturation, hypoxia-inducible factor 1α (HIF1α) a downstream modulator of hypoxia response and a proliferation marker Ki67. The reprobing method consisted of removal of the previously immunolabeled target and immunostaining with different antibodies, facilitating colocalization of enterocyte brush border epitopes as well as HIF1α, Mash1 and Prox1 in PNEC/NEB PNEC and NEBs. As these methods are suitable for routine FFPE pathology samples from various tissues, allowing visualization of multiple epitopes in the same cells/sections with superior contrast and resolution, they are suitable for a wide range of applications in diagnostic pathology and may be particularly well suited for precision medicine

  11. Conserved regions ofPlasmodium vivax potential vaccine candidate antigens in Sri Lanka:Consciousin silico analysis of prospective conformational epitope regions

    Institute of Scientific and Technical Information of China (English)

    Shanika Amarasinghe; Hashendra Kathriarachchi; Preethi Udagama

    2014-01-01

    Objective:To do mapping and modeling of conformationalB cell epitope regions of highly conserved and protective regions of three merozoitecandidate vaccine proteins ofPlasmodium vivax(P. vivax) ,ie. merozoite purface protein-1(PvMSP-1), apical membrane antigen -1 domainⅡ(PvAMA1-DⅡ) and regionⅡ of theDuffy binding protein(PvDBPⅡ), and to analyze the immunogenic properties of these predicted epitopes.Methods:3-D structures of amino acid haplotypes fromSriLanka(available inGeneBank) ofPvMSP-119(n=27),PvAMA1-DⅡ(n=21) andPvDBPⅡ(n=33) were modeled.SEPPA, selected as the best online server was used for conformational epitope predictions, while prediction and modeling of protein structure and properties related to immunogenicity was carried out withGeno3D server,SCRATCHProtein Server,NetSurfPServer and standalonesoftware,Genious5.4.4.Results:SEPPA revealed that regions of predicted conformational epitopes formed4 clusters inPvMSP-I19, and3 clusters each inPvAMA1-DⅡ andPvDBPⅡ, all of which displayed a high degree of hydrophilicity, contained solvent exposed residues, displayed high probability of antigenicity and showed positive antigenic propensity values, that indicated high degree of immunogenicity.Conclusions:Findings of this study revealed and confirmed that different parts of the sequences of each of the conserved regions of the three selected potential vaccine candidate antigens ofP. vivax are important with regard to conformational epitope prediction that warrants further laboratory experimental investigations in in vivo animal models.

  12. Implantation of a left ventricular assist device in patients with a complex apical anatomy.

    Science.gov (United States)

    Palmen, Meindert; Verwey, Harriette F; Haeck, Marlieke L A; Holman, Eduard R; Schalij, Martin J; Klautz, Robert J M

    2012-12-01

    Implantation of a left ventricular assist device can be challenging in patients with an altered apical anatomy after cardiac surgery or as the result of the presence of a calcified apical aneurysm. In this paper we present 2 cases with a challenging apical anatomy and introduce a new surgical technique facilitating left ventricular assist device implantation in these patients.

  13. Through-flow of water in leaves of a submerged plant is influenced by the apical opening

    DEFF Research Database (Denmark)

    Pedersen, Ole; Jørgensen, Lise Bolt; Sand-Jensen, Kaj

    1997-01-01

    Submerged plant, apical opening, hydathode, Sparganium, hydraulic architecture, leaf specific conductivity......Submerged plant, apical opening, hydathode, Sparganium, hydraulic architecture, leaf specific conductivity...

  14. Modelling apical constriction in epithelia using elastic shell theory.

    Science.gov (United States)

    Jones, Gareth Wyn; Chapman, S Jonathan

    2010-06-01

    Apical constriction is one of the fundamental mechanisms by which embryonic tissue is deformed, giving rise to the shape and form of the fully-developed organism. The mechanism involves a contraction of fibres embedded in the apical side of epithelial tissues, leading to an invagination or folding of the cell sheet. In this article the phenomenon is modelled mechanically by describing the epithelial sheet as an elastic shell, which contains a surface representing the continuous mesh formed from the embedded fibres. Allowing this mesh to contract, an enhanced shell theory is developed in which the stiffness and bending tensors of the shell are modified to include the fibres' stiffness, and in which the active effects of the contraction appear as body forces in the shell equilibrium equations. Numerical examples are presented at the end, including the bending of a plate and a cylindrical shell (modelling neurulation) and the invagination of a spherical shell (modelling simple gastrulation). PMID:19859751

  15. Transient apical dyskinesia with a pacemaker: Electrocardiographic features.

    Science.gov (United States)

    Núñez-Gil, Iván J; Feltes, Gisela I; Mejía-Rentería, Hernán D; Biagioni, Corina; De Agustín, J Alberto; Vivas, David; Fernández-Ortiz, Antonio

    2015-04-01

    Transient apical dyskinesia syndromes present features similar to acute coronary syndromes, but with normal coronary arteries and rapid complete resolution of wall motion alterations. We report the case of a 73-year-old woman who was admitted to hospital because of typical chest pain at rest after her brother's death. She had had a pacemaker implanted in 2001. Troponin levels were elevated and apical hypokinesia was shown by ventriculography and echocardiography, with normal coronary arteries. Evolving ECG alterations were observed in spite of the continued pacing rhythm. All these alterations were fully resolved after discharge. This case shows that, even in the presence of a pacemaker, evolving ECG alterations can be observed in Takotsubo syndrome. PMID:25840647

  16. Apical versus Basal Neurogenesis Directs Cortical Interneuron Subclass Fate

    Directory of Open Access Journals (Sweden)

    Timothy J. Petros

    2015-11-01

    Full Text Available Fate determination in the mammalian telencephalon, with its diversity of neuronal subtypes and relevance to neuropsychiatric disease, remains a critical area of study in neuroscience. Most studies investigating this topic focus on the diversity of neural progenitors within spatial and temporal domains along the lateral ventricles. Often overlooked is whether the location of neurogenesis within a fate-restricted domain is associated with, or instructive for, distinct neuronal fates. Here, we use in vivo fate mapping and the manipulation of neurogenic location to demonstrate that apical versus basal neurogenesis influences the fate determination of major subgroups of cortical interneurons derived from the subcortical telencephalon. Somatostatin-expressing interneurons arise mainly from apical divisions along the ventricular surface, whereas parvalbumin-expressing interneurons originate predominantly from basal divisions in the subventricular zone. As manipulations that shift neurogenic location alter interneuron subclass fate, these results add an additional dimension to the spatial-temporal determinants of neuronal fate determination.

  17. Apical pressures developed by needles for canal irrigation.

    Science.gov (United States)

    Bradford, C E; Eleazer, P D; Downs, K E; Scheetz, J P

    2002-04-01

    Drying instrumented canals with pressurized air may result in patient morbidity or even fatality. Low pressure and side vent needles have been suggested to lessen the danger. This study observed apical pressures from different needles inserted deeply into small round and ovoid canals as instrumentation progressed. Low-pressure (5 psi) air was injected through the needles, and apical pressures were recorded after each instrument. Pressures varied greatly within each test group. Generalities that can be drawn are that binding the needle within the canal gives higher pressures than with the needle slightly short of binding and that pressures were higher with apexes instrumented to size 30 and higher. With the needle tightly bound, neither needle size, needle design, nor canal shape resulted in statistically significant mean pressure differences. With the needle slightly withdrawn, larger bore needles gave higher pressures than small diameter needles. Caution is advised with the clinical use of pressurized air in the drying of root canals. PMID:12043877

  18. Pathogenesis of apical periodontitis and the causes of endodontic failures.

    Science.gov (United States)

    Nair, P N R

    2004-01-01

    Apical periodontitis is a sequel to endodontic infection and manifests itself as the host defense response to microbial challenge emanating from the root canal system. It is viewed as a dynamic encounter between microbial factors and host defenses at the interface between infected radicular pulp and periodontal ligament that results in local inflammation, resorption of hard tissues, destruction of other periapical tissues, and eventual formation of various histopathological categories of apical periodontitis, commonly referred to as periapical lesions. The treatment of apical periodontitis, as a disease of root canal infection, consists of eradicating microbes or substantially reducing the microbial load from the root canal and preventing re-infection by orthograde root filling. The treatment has a remarkably high degree of success. Nevertheless, endodontic treatment can fail. Most failures occur when treatment procedures, mostly of a technical nature, have not reached a satisfactory standard for the control and elimination of infection. Even when the highest standards and the most careful procedures are followed, failures still occur. This is because there are root canal regions that cannot be cleaned and obturated with existing equipments, materials, and techniques, and thus, infection can persist. In very rare cases, there are also factors located within the inflamed periapical tissue that can interfere with post-treatment healing of the lesion. The data on the biological causes of endodontic failures are recent and scattered in various journals. This communication is meant to provide a comprehensive overview of the etio-pathogenesis of apical periodontitis and the causes of failed endodontic treatments that can be visualized in radiographs as asymptomatic post-treatment periapical radiolucencies. PMID:15574679

  19. Enlargement of the apical gap after laser root resection

    Science.gov (United States)

    Mello, Guilherme P. S.; Paradella, Thais C.; Munin, Egberto; Mello, Jose B.; Pacheco, Marcos T. T.

    2000-11-01

    An apical filling material should establish, as perfect as possible, the hermetic sealing of an apical cavity. However, a gap is formed between the filling material (gutta-percha) and the root canal wall. The egress of irritants into the root canal system to the periapical tissues is considered the principal cause of fails in apicoectomy and retro-filling, being assumed that irritants penetrate mainly through the gap located between the gutta-percha and the dentin. In this paper, we report the observation of an enlargement of the apical gap, after laser apicoectomy, comparing to conventional apicoectomy. The samples were divided into groups, and the conventional apicoectomy group, together with the Er:YAG laser group (400 mJ/10 Hz) produced both similar results, being the gap unaltered. On the other hand, the samples that were irradiated with the Er:YAG laser, followed by Nd:YAG laser irradiation (1.5 W/10 Hz) presented a larger gap, conclusions that were drawn from Scanning Electronic Microscope analysis. The enlargement of the gap was due to the fusion of the dentin on the border, close to the gutta-percha. This pronounced behavior might have been caused by the surface discontinuity, imposing a non-homogeneous condition, in relation to heat propagation, existing many clinical applications of these observations.

  20. The Na+/H+ Exchanger NHE6 in the Endosomal Recycling System Is Involved in the Development of Apical Bile Canalicular Surface Domains in HepG2 Cells

    NARCIS (Netherlands)

    Ohgaki, Ryuichi; Matsushita, Masafumi; Kanazawa, Hiroshi; Ogihara, Satoshi; Hoekstra, Dick; van IJzendoorn, Sven C. D.

    2010-01-01

    Polarized epithelial cells develop and maintain distinct apical and basolateral surface domains despite a continuous flux of membranes between these domains. The Na+/H+ exchanger NHE6 localizes to endosomes but its function is unknown. Here, we demonstrate that polarized hepatoma HepG2 cells express

  1. A rapid method for the evaluation of the ionic permeabilities across epithelial cell membranes.

    Science.gov (United States)

    Movileanu, L

    1999-02-01

    This short note presents a recipe for the calculation of the ionic permeabilities across epithelial cell membranes. The method requires the Goldman-Hodgkin-Katz formalism as well as the consideration of the equivalent electrical circuit for an epithelial cell. The equivalent electrical circuit is solved in terms of the equivalent electromotive forces coupled in series with the ionic resistances of both cell membranes (apical and basolateral). The present procedure is feasible for any leaky epithelial cell membrane with the condition that this membrane (apical or basolateral) does not contain primary or secondary mechanisms for active transport. PMID:10100952

  2. Calcium hydroxide induced apical barrier in fractured nonvital immature permanent incisors

    Directory of Open Access Journals (Sweden)

    Vellore K

    2010-06-01

    Full Text Available Management of trauma to an immature permanent incisor is a challenge, especially when the pulp is necrotic. The aim of this study is to discuss the management of fractured maxillary right and left central incisors with immature root apex. Radiographic evaluation revealed that the tooth 11 had convergent apical walls, whereas tooth 21 had parallel apical walls; both the incisors were treated with pure calcium hydroxide paste to induce apical development. Follow-up clinical and radiographic examinations confirmed apical barrier at 3 months. Radiograph of tooth 11 with convergent roots showed continued apical development, whereas tooth 21 showed apical bridging, following which root canal was completed in both the teeth. Therefore, in view of the simplicity of this method, we believe that calcium hydroxide can be reemphasized as one of the treatment alternatives for fractured nonvital immature permanent incisors to induce apical barrier.

  3. Antigen processing and remodeling of the endosomal pathway: requirements for antigen cross-presentation.

    Directory of Open Access Journals (Sweden)

    Ewoud Bernardus Compeer

    2012-03-01

    Full Text Available The cross-presentation of endocytosed antigen as peptide/class I MHC complexes plays a central role in the elicitation of CD8+ T cell clones that mediate anti-viral and anti-tumor immune responses. While it has been clear that there are specific subsets of professional antigen presenting cells (APC capable of antigen cross-presentation, description of mechanisms involved is still ongoing. Especially amongst dendritic cells (DC, there are specialized subsets that are highly proficient at antigen cross-presentation. We here present a focused survey on the cell biological processes in the endosomal pathway that support antigen cross-presentation. This review highlight DC-intrinsic mechanisms that facilitate the cross-presentation of endocytosed antigen, including receptor-mediated uptake, recycling and maturation including the sorting of membrane proteins, dynamic remodeling of endosomal structures and cell-surface directed endosomal trafficking. We will conclude with description of pathogen-induced deviation of endosomal processing, and discuss how immune evasion strategies pertaining endosomal trafficking may preclude antigen cross-presentation.

  4. Occurrence of the Transition of Apical Architecture and Expression Patterns of Related Genes during Conversion of Apical Meristem Identity in G2 Pea

    Institute of Scientific and Technical Information of China (English)

    Da-Yong Wang; Qing Li; Ke-Ming Cui; Yu-Xian Zhu

    2009-01-01

    G2 pea exhibits an apical senescence delaying phenotype under short-day (SD) conditions; however, the structural basis for its apical development is still largely unknown. In the present study, the apical meristem of SD-grown G2 pea plants underwent a transition from vegetative to indeterminate inflorescence meristem, but the apical meristem of long-day (LD)-grown G2 pea plants would be further converted to determinate floral meristem. Both SD signal and GA3 treatment enhanced expression of the putative calcium transporter PPF1, and pea homologs of TFL1 (LF and DET), whereas LD signal suppressed their expression at 60 d post-flowering compared with those at 40 d post-flowering. Both PPF1 and LF expressed at the vegetative and reproductive phases in SD-grown apical buds, but floral initiation obviously increased the expression level of PPF1 compared with the unchanged expression level of LF from 40 to 60 d post-flowering. In addition, although the floral initiation significantly enhanced the expression levels of PPF1 and DET, DET was mainly expressed after floral initiation in SD-grown apical buds. Therefore, the main structural difference between LD- and SD-grown apical meristem in G2 pea lies in whether their apical indeterminate inflorescence medstem could be converted to the determinate structure.

  5. SEM investigation of Er:YAG laser apical preparation

    Science.gov (United States)

    Bǎlǎbuc, Cosmin; Todea, Carmen; Locovei, Cosmin; RǎduÅ£ǎ, Aurel

    2016-03-01

    Endodontic surgery involves the incision and flap elevation, the access to the root tip, its resection, the cavity retrograde preparation and filling it with biocompatible material that provides a good seal of the apex[1]. Apicoectomy is compulsory in endodontic surgery. The final stage involves the root retropreparation and the carrying out of the retrograde obturation. In order to perform the retrograde preparation the endodontist can use various tools such as lowspeed conventional handpieces, sonic and ultrasonic equipment. The ideal depth of the preparation should be 3 mm, exceeding this value may affect the long-term success of the obturation [2]. Resection at the depth of 3 mm reduces apical ramifications by 98% and lateral root canals by 93%. The ultrasonic retropreparation has numerous advantages compared to the dental drill. Firstly, the cavity will be in the axis of the tooth which implies a minimum destruction of the root canal morphology. The preparations are precise, and the cutting pattern is perpendicular to the long axis of the root, the advantage being the reduction in the number of dentinal tubules exposed at the resected area [3]. Therefore, the retrograde filling is the procedure when an inert and non-toxic material is compacted in the apically created cavity.[4,5]. The Er:YAG laser is the most common wavelength indicated for dental hard tissue preparation. Its natural selectivity offers a significant advantage compared to the conventional hard tissue preparation [6-9].The purpose of this in vitro study was to investigate the quality of Er:YAG laser apical third preparation using Scanning Electron Microscopy (SEM), in comparison with the conventional ultrasonic method.

  6. Selamento apical proporcionado por diferentes cimentos endodônticos

    Directory of Open Access Journals (Sweden)

    Karlel Tristão Marques

    2011-01-01

    Full Text Available To have success in endodontic treatment it is necessary that the sealing airtight as possible of the root canal system, with suitable material such as gutta-percha and sealer. There are sealers of different compositions, which should allow the sealing of the root canal system, preventing recontamination place and favoring the success of endodontic therapy. In this study were evaluated the apical microleakage after obturation using different root canal sealers widely employed in the market, with different compositions. For this study, 85 single rooted teeth premolars which have removed their crowns and root canals prepared and filled using four types of cement: AH Plus, Endofill, Acroseal and Real Seal SE. All samples were sealed and submitted to microleakage with Indian ink; longitudinal cuts made on both sides of the tooth and cleaved; apical images were obtained. The areas of dye penetration were measured on a computer using specific software and subjected to statistical analysis. It was concluded that the Acroseal ® showed the highest mean microleakage with no statistically significant difference in relation to the RealSeal SE®, but with significant differences compared to Endofill ® and AHPlus ®.

  7. Calcium hydroxide as intracanal dressing for teeth with apical periodontitis

    Directory of Open Access Journals (Sweden)

    Sari Dewiyani

    2011-03-01

    Full Text Available Background: Root canal infection and periapical diseases are caused by bacteria and their products. Long term infection may spread bacteria throughout the root canal system. Apical periodontitis caused by infectious microbe that persistent in root canals can cause radiographic and histopathology periapical changes. Chemomechanical preparation and intracanal dressing then are recommended to be conducted and used in between visits to eliminate microbes in root canals. Calcium hydroxide (Ca(OH2 can be used as intracanal dressing since it can be used as musical physical defense barrier to eliminate re-infection in root canal and to disturb nutrition supply for bacterial development. Purpose: The aim of this study is observe the effectiveness of Ca(OH2 in treating endodontic teeth with apical periodontitis. Cases: Case 1 and 3 are about patients whose left posterior mandibular teeth had spontaneous intermittent pain. Case 2 is about a patient whose left posterior maxillary teeth had gingival abscess and fracture history. Based on the radiographic examination, it was known that the filling of root canal was incomplete and there was radiolucency in the apical area. Case management: The cases were treated with triad endodontics, which involves preparation, disinfection by using 2.5% NaOCl as irrigation substance and calcium hydroxide as intracanal dressing, and then the filling of root canal with gutta percha and endomethasone root canal cement. Evaluations were conducted one month, 12 months, and 24 months after the treatment. Conclusion: Calcium hydroxide is effective to be used as intracanal dressing in apical periodontitis cases.Latar belakang: Infeksi saluran akar dan penyakit periapeks disebabkan oleh mikroba dan produknya. Infeksi yang berlangsung lama memungkinkan bakteri masuk ke dalam seluruh sistem saluran akar. Periodontitis apikal disebabkan oleh infeksi persisten mikroba di dalam sistem saluran akar disertai perubahan radiografik dan

  8. Prognostic factors in apical surgery with root-end filling: a meta-analysis

    DEFF Research Database (Denmark)

    von Arx, Thomas; Peñarrocha, Miguel; Jensen, Storgård

    2010-01-01

    Apical surgery has seen continuous development with regard to equipment and surgical technique. However, there is still a shortage of evidence-based information regarding healing determinants. The objective of this meta-analysis was to review clinical articles on apical surgery with root-end fill......Apical surgery has seen continuous development with regard to equipment and surgical technique. However, there is still a shortage of evidence-based information regarding healing determinants. The objective of this meta-analysis was to review clinical articles on apical surgery with root...

  9. com e sem remoção do delta apical

    Directory of Open Access Journals (Sweden)

    Guilherme Mello

    2006-01-01

    Full Text Available This study evaluated the apex cut and apical fusion by Nd:YAG laser at apical surgery. 24 human canines were endodontic treated and randomly divided into two groups. In A group the apicoectomy was performed at 90 degrees and 3mm from the apex extreme; in B group the apex was kept. Both groups were irradiated by Nd:YAG laser (10Hz, 150mJ, 60s. The results showed less infiltration to A group (p<0,001. When Nd:YAG laser apical fusion after apicoectomy were performed at the parameters of this study, best apical isolation is obtained.

  10. Membrane recycling at the infranuclear pole of the outer hair cell

    Science.gov (United States)

    Harasztosi, Csaba; Harasztosi, Emese; Gummer, Anthony W.

    2015-12-01

    Rapid endocytic activity of outer hair cells (OHCs) in the guinea-pig cochlea has been already studied using the fluorescent membrane marker FM1-43. It was demonstrated that vesicles were endocytosed at the apical pole of OHCs and transcytosed to the basolateral membrane and through a central strand towards the nucleus. The significance of endocytic activity in the infranuclear region is still not clear. Therefore, in this study endocytic activity at the synaptic pole of OHCs was investigated. Confocal laser scanning microscopy was used to visualize dye uptake of OHCs isolated from the guinea-pig cochlea. Signal intensity changes were quantified in the apical and basal poles relative to the signal at the membrane. Data showed no significant difference in fluorescent signal intensity changes between the opposite poles of the OHC. These results suggest that endocytic activities in both the basal and the apical poles contribute equally to the membrane recycling of OHCs.

  11. MAL Is a Regulator of the Recruitment of Myelin Protein PLP to Membrane Microdomains

    NARCIS (Netherlands)

    Bijlard, Marjolein; de Jonge, Jenny C.; Klunder, Bert; Nomden, Anita; Hoekstra, Dick; Baron, Wia

    2016-01-01

    In oligodendrocytes (OLGs), an indirect, transcytotic pathway is mediating transport of de novo synthesized PLP, a major myelin specific protein, from the apical-like plasma membrane to the specialized basolateral-like myelin membrane to prevent its premature compaction. MAL is a well-known regulato

  12. Magi Is Associated with the Par Complex and Functions Antagonistically with Bazooka to Regulate the Apical Polarity Complex

    Science.gov (United States)

    Padash Barmchi, Mojgan; Samarasekera, Gayathri; Gilbert, Mary; Auld, Vanessa J.; Zhang, Bing

    2016-01-01

    The mammalian MAGI proteins play important roles in the maintenance of adherens and tight junctions. The MAGI family of proteins contains modular domains such as WW and PDZ domains necessary for scaffolding of membrane receptors and intracellular signaling components. Loss of MAGI leads to reduced junction stability while overexpression of MAGI can lead to increased adhesion and stabilization of epithelial morphology. However, how Magi regulates junction assembly in epithelia is largely unknown. We investigated the single Drosophila homologue of Magi to study the in vivo role of Magi in epithelial development. Magi is localized at the adherens junction and forms a complex with the polarity proteins, Par3/Bazooka and aPKC. We generated a Magi null mutant and found that Magi null mutants were viable with no detectable morphological defects even though the Magi protein is highly conserved with vertebrate Magi homologues. However, overexpression of Magi resulted in the displacement of Baz/Par3 and aPKC and lead to an increase in the level of PIP3. Interestingly, we found that Magi and Baz functioned in an antagonistic manner to regulate the localization of the apical polarity complex. Maintaining the balance between the level of Magi and Baz is an important determinant of the levels and localization of apical polarity complex. PMID:27074039

  13. Cryopreservation of Galanthus elwesii Hook. apical meristems by droplet vitrification.

    Science.gov (United States)

    Maslanka, M; Panis, B; Bach, A

    2013-01-01

    The aim of this study was to develop an efficient cryopreservation protocol for the geophyte giant snowdrop (Galanthus elwesii Hook.) that guarantees a high rate of survival and plant regeneration after cryopreservation. The excised apical meristems were obtained from cultures of in vitro grown bulb scales. Using a vitrification procedure and optimizing the duration of the exposure to the loading solution (LS), meristem post-rewarm survival rates higher than 90 percent were achieved. Also regrowth percentages were very high, ranging from 87 to 91 percent. After optimizing the time of exposure to the plant vitrification solution (PVS2), the survival rate was between 83 and 97 percent. During post-rewarm regeneration, good growth recovery was as high as 76 percent; however, hyperhydration and callusing were also observed. The results demonstrate that cryopreservation of Galanthus elwesii germplasm seems to be feasible. PMID:23435704

  14. Case of cheilitis granulomatosa associated with apical periodontitis.

    Science.gov (United States)

    Kawakami, Tomoko; Fukai, Kazuyoshi; Sowa, Junko; Ishii, Masamitsu; Teramae, Hiroyuki; Kanazawa, Koutetsu

    2008-02-01

    The etiology of cheilitis granulomatosa is unknown. In some cases, rapid improvement and/or complete elimination of swelling of the lips after dental treatment has been reported. Here, we describe another case of improvement following dental treatment. A 57-year-old woman had developed asymptomatic swelling of the lower lip 2 months previously. Histological examination revealed non-caseous giant cell granulomas. Neither facial nerve palsy nor fissuring of the tongue was present. Patch testing for metal allergy revealed only mild irritation to zinc ion. Although topical corticosteroid ointment and oral tranilast for 4 months were ineffective, rapid and remarkable improvement of the swelling was noted soon after treatment of two lesions of apical periodontitis. Thorough examination for foci of infection is necessary when treating a patient with cheilitis granulomatosa. PMID:18271809

  15. Cryopreservation of Galanthus elwesii Hook. apical meristems by droplet vitrification.

    Science.gov (United States)

    Maslanka, M; Panis, B; Bach, A

    2013-01-01

    The aim of this study was to develop an efficient cryopreservation protocol for the geophyte giant snowdrop (Galanthus elwesii Hook.) that guarantees a high rate of survival and plant regeneration after cryopreservation. The excised apical meristems were obtained from cultures of in vitro grown bulb scales. Using a vitrification procedure and optimizing the duration of the exposure to the loading solution (LS), meristem post-rewarm survival rates higher than 90 percent were achieved. Also regrowth percentages were very high, ranging from 87 to 91 percent. After optimizing the time of exposure to the plant vitrification solution (PVS2), the survival rate was between 83 and 97 percent. During post-rewarm regeneration, good growth recovery was as high as 76 percent; however, hyperhydration and callusing were also observed. The results demonstrate that cryopreservation of Galanthus elwesii germplasm seems to be feasible.

  16. Serious arrhythmias in patients with apical hypertrophic cardiomyopathy

    International Nuclear Information System (INIS)

    We report cases of serious arrhythmias associated with apical hypertrophic cardiomyopathy (AHCM). Thirty-one patients were referred to our institute to undergo further assessment of their AHCM from 1988 to 1999. Three patients with nonsustained ventricular tachycardia demonstrated an 123I-MIBG regional reduction in the tracer uptake. In two patients with ventricular fibrillation (VF), the findings from 123I-MIBG imaging revealed regional sympathetic denervation in the inferior and lateral regions. Electrophysiologic study demonstrated reproducible induction of VF in aborted sudden death and presyncopal patients, resulting in the need for an implantable defibrillator device and amiodarone in each patient. Patients with refractory atrial fibrillation with a rapid ventricular response suffered from serious congestive heart failure. A prudent assessment and strategy in patients with this disease would be indispensable in avoiding a disastrous outcome. (author)

  17. Serious arrhythmias in patients with apical hypertrophic cardiomyopathy

    Energy Technology Data Exchange (ETDEWEB)

    Okishige, Kaoru; Sasano, Tetsuo; Yano, Kei; Azegami, Kouji; Suzuki, Kou; Itoh, Kuniyasu [Yokohama Red Cross Hospital (Japan)

    2001-05-01

    We report cases of serious arrhythmias associated with apical hypertrophic cardiomyopathy (AHCM). Thirty-one patients were referred to our institute to undergo further assessment of their AHCM from 1988 to 1999. Three patients with nonsustained ventricular tachycardia demonstrated an {sup 123}I-MIBG regional reduction in the tracer uptake. In two patients with ventricular fibrillation (VF), the findings from {sup 123}I-MIBG imaging revealed regional sympathetic denervation in the inferior and lateral regions. Electrophysiologic study demonstrated reproducible induction of VF in aborted sudden death and presyncopal patients, resulting in the need for an implantable defibrillator device and amiodarone in each patient. Patients with refractory atrial fibrillation with a rapid ventricular response suffered from serious congestive heart failure. A prudent assessment and strategy in patients with this disease would be indispensable in avoiding a disastrous outcome. (author)

  18. Molecular cloning of complementary DNAs encoding the heavy chain of the human 4F2 cell-surface antigen: a type II membrane glycoprotein involved in normal and neoplastic cell growth

    International Nuclear Information System (INIS)

    Complementary DNA (cDNA) clones encoding the heavy chain of the heterodimeric human membrane glycoprotein 4F2 have been isolated by immunoscreening of a λgt11 expression library. The identity of these clones has been confirmed by hybridization to RNA and DNA prepared from mouse L-cell transfectants, which were produced by whole cell gene transfer and selected for cell-surface expression of the human 4F2 heavy chain. DNA sequence analysis suggest that the 4F2 heavy-chain cDNAs encode an approximately 526-amino acid type II membrane glycoprotein, which is composed of a large C-terminal extracellular domain, a single potential transmembrane region, and a 50-81 amino acid N-terminal intracytoplasmic domain. Southern blotting experiments have shown that the 4F2 heavy-chain cDNAs are derived from a single-copy gene that has been highly conserved during mammalian evolution

  19. Clonging and Expression of Leptospiral Outer Membrane Protein LipL32 Gene and Application of Recombinant Antigen in Enzyme-linked Immunosorbent Assays%钩端螺旋体外膜脂蛋白LipL32基因的克隆和表达及其在ELISA检测中的应用

    Institute of Scientific and Technical Information of China (English)

    范薇; 于长明; 杨敬; 隋丽华; 战大伟; 贺争鸣; 孙岩松

    2003-01-01

    Objective To construct L32-pQE32 recombinant expression vectors, and to induce the expression of recombinant Leptospiral outer membrane protein LipL32. Establish method of recombinant Leptospiral outer membrane proteinbased ELISA. Method Gene coding of Leptospiral LipL32 protein was amplified by PCR, then recombinant cloning vectors pGEM-T/L32 and expression vectors L32-pQE32 were constructed. Recombinant expression vector was transformed into the competent host E. coli. DH-5α and E. coli. M15. Recombinant Leptospiral LipL32 protein was expressed by IPTG induced method. Immulon microtiter plates were coated at 37℃ overnight with 100 ng of purified recombinant protein per well, 3 positive and 4 negative sera were used in indirect ELISA. Results Mature Leptospiral LipL32 gene fragment about 750 bp was amplified by PCR. LipL32 gene was inserted into expression vectors pQE32, the molecular weight of fusion protein was corresponding to the estimated molecular size of mature Leptospiral LipL32 protein. Results of Western-blot and ELISA demonstrated intense LipL32 reactivity with anti-Leptospira sera. Conclusion findings indicate that recombinant Leptospiral LipL32 may be an important, useful antigen for the serodiagnosis of Leptospira.

  20. Oral microbiota species in acute apical endodontic abscesses

    Science.gov (United States)

    George, Noelle; Flamiatos, Erin; Kawasaki, Kellie; Kim, Namgu; Carriere, Charles; Phan, Brian; Joseph, Raphael; Strauss, Shay; Kohli, Richie; Choi, Dongseok; Craig Baumgartner, J.; Sedgley, Christine; Maier, Tom; Machida, Curtis A.

    2016-01-01

    Background and objectives Acute apical abscesses are serious endodontic diseases resulting from pulpal infection with opportunistic oral microorganisms. The objective of this study was to identify and compare the oral microbiota in patients (N=18) exhibiting acute apical abscesses, originating from the demographic region in Portland, Oregon. The study hypothesis is that abscesses obtained from this demographic region may contain unique microorganisms not identified in specimens from other regions. Design Endodontic abscesses were sampled from patients at the Oregon Health & Science University (OHSU) School of Dentistry. DNA from abscess specimens was subjected to polymerase chain reaction amplification using 16S rRNA gene-specific primers and Cy3-dCTP labeling. Labeled DNA was then applied to microbial microarrays (280 species) generated by the Human Oral Microbial Identification Microarray Laboratory (Forsyth Institute, Cambridge, MA). Results The most prevalent microorganisms, found across multiple abscess specimens, include Fusobacterium nucleatum, Parvimonas micra, Megasphaera species clone CS025, Prevotella multisaccharivorax, Atopobium rimae, and Porphyromonas endodontalis. The most abundant microorganisms, found in highest numbers within individual abscesses, include F. nucleatum, P. micra, Streptococcus Cluster III, Solobacterium moorei, Streptococcus constellatus, and Porphyromonas endodontalis. Strong bacterial associations were identified between Prevotella multisaccharivorax, Acidaminococcaceae species clone DM071, Megasphaera species clone CS025, Actinomyces species clone EP053, and Streptococcus cristatus (all with Spearman coefficients >0.9). Conclusions Cultivable and uncultivable bacterial species have been identified in endodontic abscesses obtained from the Portland, Oregon demographic region, and taxa identifications correlated well with other published studies, with the exception of Treponema and Streptococcus cristae, which were not commonly

  1. A Role For Mitochondria In Antigen Processing And Presentation.

    Science.gov (United States)

    Bonifaz, Lc; Cervantes-Silva, Mp; Ontiveros-Dotor, E; López-Villegas, Eo; Sánchez-García, Fj

    2014-09-23

    Immune synapse formation is critical for T lymphocyte activation, and mitochondria have a role in this process, by localizing close to the immune synapse, regulating intracellular calcium concentration, and providing locally required ATP. The interaction between antigen presenting cells (APCs) and T lymphocytes is a two-way signaling process. However, the role of mitochondria in antigen presenting cells during this process remains unknown. For APCs to be able to activate T lymphocytes, they must first engage in an antigen-uptake, -processing, and -presentation process. Here we show that HEL-loaded B lymphocytes, as a type of APCs, undergo a small but significant mitochondrial depolarization by 1-2 h following antigen exposure thus suggesting an increase in their metabolic demands. Inhibition of ATP synthase (oligomycin) or mitochondrial Ca(2+) uniporter (MCU) (Ruthenium red) had no effect on antigen uptake. Therefore, antigen processing and antigen presentation were further analyzed. Oligomycin treatment reduced the amount of specific MHC-peptide complexes but not total MHC II on the cell membrane of B lymphocytes which correlated with a decrease in antigen presentation. However, oligomycin also reduced antigen presentation by B lymphocytes that endogenously express HEL and by B lymphocytes loaded with the HEL48-62 peptide, although to a lesser extent. ATP synthase inhibition and MCU inhibition had a clear inhibitory effect on antigen processing (DQ-OVA). Taking together these results suggest that ATP synthase and MCU are relevant for antigen processing and presentation. Finally, APCs mitochondria were found to re-organize towards the APC-T immune synapse. This article is protected by copyright. All rights reserved.

  2. Mechanisms of apical-basal axis orientation and epithelial lumen positioning

    NARCIS (Netherlands)

    Overeem, Arend W.; Bryant, David M.; van IJzendoorn, Sven C. D.

    2015-01-01

    In epithelial cells, the polarized orientation of the apical-basal axis determines the position of the apical lumen and, thereby, the collective tubular tissue architecture. From recent studies employing 3D cell cultures, animal models, and patient material, a model is emerging in which the orientat

  3. Developmental Corneal Innervation: Interactions between Nerves and Specialized Apical Corneal Epithelial Cells

    OpenAIRE

    Kubilus, James K.; Linsenmayer, Thomas F.

    2010-01-01

    During developmental innervation of the chicken cornea, nerves interact with apical corneal epithelial cells to form synapse-like structures. In addition, these apical epithelial cells express class III β-tubulin, an isoform of β-tubulin generally thought to be neuron specific.

  4. Unpolarized release of vaccinia virus and HIV antigen by colchicine treatment enhances intranasal HIV antigen expression and mucosal humoral responses.

    Directory of Open Access Journals (Sweden)

    Yan Zhang

    Full Text Available The induction of a strong mucosal immune response is essential to building successful HIV vaccines. Highly attenuated recombinant HIV vaccinia virus can be administered mucosally, but even high doses of immunization have been found unable to induce strong mucosal antibody responses. In order to solve this problem, we studied the interactions of recombinant HIV vaccinia virus Tiantan strain (rVTT-gagpol in mucosal epithelial cells (specifically Caco-2 cell layers and in BALB/c mice. We evaluated the impact of this virus on HIV antigen delivery and specific immune responses. The results demonstrated that rVTT-gagpol was able to infect Caco-2 cell layers and both the nasal and lung epithelia in BALB/c mice. The progeny viruses and expressed p24 were released mainly from apical surfaces. In BALB/c mice, the infection was limited to the respiratory system and was not observed in the blood. This showed that polarized distribution limited antigen delivery into the whole body and thus limited immune response. To see if this could be improved upon, we stimulated unpolarized budding of the virus and HIV antigens by treating both Caco-2 cells and BALB/c mice with colchicine. We found that, in BALB/c mice, the degree of infection and antigen expression in the epithelia went up. As a result, specific immune responses increased correspondingly. Together, these data suggest that polarized budding limits antigen delivery and immune responses, but unpolarized distribution can increase antigen expression and delivery and thus enhance specific immune responses. This conclusion can be used to optimize mucosal HIV vaccine strategies.

  5. Effect of master apical file size and taper on irrigation and cleaning of the apical third of curved canals.

    Directory of Open Access Journals (Sweden)

    Nahid Mohammadzadeh Akhlaghi

    2014-04-01

    Full Text Available Canal preparation generates a substantial amount of debris and smear layer (SL. The size and taper of the Master Apical File (MAF affects on penetration of irrigants and subsequently canal cleaning efficacy. The aim of this study was to evaluate the effect of MAF size and taper on penetration of irrigants to the apical third of curved mesiobuccal (MB canals of mandibular first molars.Eighty-nine human mandibular first molars were divided into one control group (n=5 without rotary instrumentation and 6 experimental groups (n=14 each that were prepared with the following RaCe rotary files as MAF: 25.04 (group1, 25.06 (group 2, 30.04 (group 3, 30.06 (group 4, 35.04 (group 5 and 35.06 (group 6. All the experimental groups were rinsed with 2 ml of 17 % EDTA followed by 2 ml of 5.25% NaOCl. Debridement of the MB canals was evaluated using scanning electron microscope (SEM. The data were statistically analyzed using Kruskal-Wallis and Mann-Whitney U tests (P<0.05.Group 6 (MAF=35.06 showed 100% acceptable debridement. This rate was 92.9% for MAF=35.04. In group 4 (MAF=30.06 smear layer (SL was removed in the three-forth of the samples and debris was removed in 92.9% of them. Acceptable debridement was not achieved in most samples of groups 1 and 2 (25.04 and 25.06, respectively and the mentioned two groups had statistically significant difference in this respect with the other groups (P<0.05.Based on this study, 30.06 may be considered as the minimum MAF size for acceptable debridement.

  6. The role of CD4 in antigen-independent activation of isolated single T lymphocytes

    DEFF Research Database (Denmark)

    Kelso, A; Owens, T

    1988-01-01

    The membrane molecule CD4 (L3T4) is thought to facilitate activation of Class II H-2-restricted T cells by binding to Ia determinants on antigen-presenting cells. Recent reports suggest that CD4 can also contribute to antigen-independent activation by anti-T cell receptor (TCR) antibodies. An assay...

  7. Assessment of Root Morphology and Apices of First and Second Maxillary Molars in Tehran Population

    Directory of Open Access Journals (Sweden)

    Mandana Naseri

    2015-12-01

    Full Text Available Introduction: Objective: This study aimed to assess the possible variations in root canal anatomy and topography of the apices of first and second maxillary molars. Materials and methods: A total of 67 first and second maxillary permanent molars were collected. Access cavity was prepared and 2% methylene blue was injected. The teeth were demineralized by 5% nitric acid and cleared with methyl salicylate. Specimens were evaluated under stereomicroscopy and analyzed using the sample t-test. Results: Based on Vertucci’s classification, the mesiobuccal root of maxillary first molars was type I in 87.5% and type IV in 12.5% of the cases. The mesiobuccal root of second maxillary molars was type I in 60%, type II in 8.6%, type IV in 25.7% and type V in 5.7% of cases. In maxillary first and second molars, the distobuccal and palatal roots were type I in 100% of the cases. The distance of the apical constriction from the apical foramen was 0.21±0.09 mm, the distance from the apical constriction tothe anatomic apex was 0.44±0.19 mm and the distance of the apical foramen from the anatomic apex was 0.15±0.15 mm. The mean percentage of delta prevalence was 3.2% in both teeth. Conclusion: The mean distance of the apical foramen and apical constriction from the anatomic apex was less than 0.6 and 1.2 mm, respectively. In maxillary first and second molars, the mean distance of the apical constriction from the apical foramen and anatomic apex was 0.21 and 0.44, respectively and the mean distance of the apical foramen from the anatomic apex was 0.15 mm

  8. Evaluation of apical subtype of hypertrophic cardiomyopathy using cardiac magnetic resonance imaging with gadolinium enhancement.

    Science.gov (United States)

    Kebed, Kalie Y; Al Adham, Raed I; Bishu, Kalkidan; Askew, J Wells; Klarich, Kyle W; Araoz, Philip A; Foley, Thomas A; Glockner, James F; Nishimura, Rick A; Anavekar, Nandan S

    2014-09-01

    Apical hypertrophic cardiomyopathy (HC) is an uncommon variant of HC. We sought to characterize cardiac magnetic resonance imaging (MRI) findings among apical HC patients. This was a retrospective review of consecutive patients with a diagnosis of apical HC who underwent cardiac MRI examinations at the Mayo Clinic (Rochester, MN) from August 1999 to October 2011. Clinical and demographic data at the time of cardiac MRI study were abstracted. Cardiac MRI study and 2-dimensional echocardiograms performed within 6 months of the cardiac MRI were reviewed; 96 patients with apical HC underwent cardiac MRI examinations. LV end-diastolic and end-systolic volumes were 130.7 ± 39.1 ml and 44.2 ± 20.9 ml, respectively. Maximum LV thickness was 19 ± 5 mm. Hypertrophy extended beyond the apex into other segments in 57 (59.4%) patients. Obstructive physiology was seen in 12 (12.5%) and was more common in the mixed apical phenotype than the pure apical (19.3 vs 2.6%, p = 0.02). Apical pouches were noted in 39 (40.6%) patients. Late gadolinium enhancement (LGE) was present in 70 (74.5%) patients. LGE was associated with severe symptoms and increased maximal LV wall thickness. In conclusion, cardiac MRI is well suited for studying the apical form of HC because of difficulty imaging the cardiac apex with standard echocardiography. Cardiac MRI is uniquely suited to delineate the presence or absence of an apical pouch and abnormal myocardial LGE that may have implications in the natural history of apical HM. In particular, the presence of abnormal LGE is associated with clinical symptoms and increased wall thickness. PMID:25037678

  9. Membrane dynamics

    DEFF Research Database (Denmark)

    Bendix, Pól Martin

    2015-01-01

    Current topics include membrane-protein interactions with regard to membrane deformation or curvature sensing by BAR domains. Also, we study the dynamics of membrane tubes of both cells and simple model membrane tubes. Finally, we study membrane phase behavior which has important implications for...

  10. 钩端螺旋体外膜蛋白 Loa22优势 T-B联合抗原表位及其免疫原性研究%Identification and immunogenicity analysis of predominant T-B combined antigenic epitopes on the ;outer membrane protein Loa22 of Leptospira interrogans strains

    Institute of Scientific and Technical Information of China (English)

    阮萍; 赵金方; 李阳; 严杰; 胡玮琳

    2015-01-01

    目的:筛选并鉴定致病性问号钩端螺旋体(简称钩体)外膜蛋白Loa22优势T细胞和B细胞( T/B)联合抗原表位及其免疫原性。方法采用PCR检测我国流行的8群8型株问号钩体loa22基因,T-A克隆后测序。构建问号钩体黄疸出血群赖型赖株loa22基因原核表达系统。 Ni-NTA亲和层析法提纯表达的目的重组蛋白rLoa22并制备其兔抗血清及其IgG。采用生物信息学软件预测Loa22的T-B联合抗原表位。采用噬菌体展示联合Western blot法、ELISA分别检测重组噬菌体PⅢ蛋白展示的T-B联合表位肽和人工合成T-B联合表位肽的免疫原性。采用MTS法和ELISA分别检测T-B联合表位肽诱导T细胞活化及其分泌IL-2、IL-4和IFN-γ情况。结果所有受检的致病性钩体株均能检出loa22基因,其核苷酸和氨基酸序列相似性高达85.5%~99.8%和93.9%~99.5%。所构建的loa22基因原核表达系统能高效表达rLoa22。 Loa22-77、Loa22-90、Loa22-125和Loa22-157这4个T-B联合抗原表位中,仅有Loa22-90显示了很强的Western blot阳性条带。 Loa22-90能有效诱导CD4+T细胞增殖及IL-2(Th1)和IL-4(Th2)水平显著升高(P<0.05)。结论 Loa22是问号钩体序列保守的属特异性外膜蛋白抗原,其优势T-B联合抗原表位为Loa22-90,该表位可作为钩端螺旋体多抗原肽疫苗的候选表位。%Objective To screen and identify the predominant T-and B-cell ( T-B) combined an-tigenic epitopes on the outer membrane protein Loa22 of pathogenic Leptospira interrogans ( L.interrogans) stains and to further analyze their immunogenicity.Methods PCR analysis was used to detect loa22 gene in L.interrogans strains belonging to eight different serogroups or serovars prevalent in China.The PCR prod-ucts were sequenced after T-A cloning.A prokaryotic expression system for loa22 gene of L.interrogans sero-group Icterohaemorrhagiae serovar Lai strain Lai was

  11. AMP-activated protein kinase inhibits alkaline pH- and PKA-induced apical vacuolar H+-ATPase accumulation in epididymal clear cells

    OpenAIRE

    Hallows, Kenneth R.; Alzamora, Rodrigo; Li, Hui; Gong, Fan; Smolak, Christy; Neumann, Dietbert; Pastor-Soler, Núria M.

    2009-01-01

    Acidic luminal pH and low [HCO3−] maintain sperm quiescent during maturation in the epididymis. The vacuolar H+-ATPase (V-ATPase) in clear cells is a major contributor to epididymal luminal acidification. We have shown previously that protein kinase A (PKA), acting downstream of soluble adenylyl cyclase stimulation by alkaline luminal pH or HCO3−, induces V-ATPase apical membrane accumulation in clear cells. Here we examined whether the metabolic sensor AMP-activated protein kinase (AMPK) reg...

  12. Comparison of apical sealing and periapical extrusion of the ThermaFil obturation technique with and without MTA as an apical barrier: An in vitro study

    Directory of Open Access Journals (Sweden)

    Satish Kumar

    2011-01-01

    Full Text Available Aim: The aim of the study was to compare the conventional ThermaFil obturation technique and ThermaFil obturation with mineral trioxide aggregate (MTA as an apical barrier, with regard to apical sealing and extrusion. Materials and Methods: Twenty extracted human canines were instrumented using a crown-down technique and divided into two groups. The experimental group was obturated using ThermaFil obturation with MTA as an apical barrier and the control group was obturated using the conventional ThermaFil obturation technique. AH Plus sealer was used in both the groups. Apical extrusion was recorded. Teeth of both the groups were coated with nail polish, except for the apical 3 mm. After 24 h, they were suspended in black India ink for 48 h. Canines were decalcified, rendered transparent, and linear dye penetration was measured under ×40 stereomicroscope. Results: There was a significant extrusion noticed in conventional ThermaFil obturation technique. Frequency of extrusion of sealer and/or gutta-percha was supposed to be evaluated using χ² test, but since the values of the samples of ThermaFil plus MTA group were zero, statistical analysis could not be conducted, whereas linear dye leakage was calculated with Mann-Whitney U test because the distribution was abnormal. Conclusion: Although ThermaFil plus MTA group showed microleakage, extrusion of sealer and the core material was prevented in comparison with conventional ThermaFil obturation technique. It is advantageous to use MTA as an apical plug as there is no fear of apical extrusion and the root canal system can then be packed three dimensionally against this barrier using any thermoplasticized gutta-percha obturation technique.

  13. Pattern of distribution of blood group antigens on human epidermal cells during maturation

    DEFF Research Database (Denmark)

    Dabelsteen, Erik; Buschard, Karsten; Hakomori, Sen-Itiroh

    1984-01-01

    The distribution in human epidermis of A, B, and H blood group antigens and of a precursor carbohydrate chain, N-acetyl-lactosamine, was examined using immunofluorescence staining techniques. The material included tissue from 10 blood group A, 4 blood group B, and 9 blood group O persons. Murine...... on the lower spinous cells whereas H antigen was seen predominantly on upper spinous cells or on the granular cells. Epithelia from blood group A or B persons demonstrated A or B antigens, respectively, but only if the tissue sections were trypsinized before staining. In such cases A or B antigens were found...... monoclonal antibodies were used to identify H antigen (type 2 chain) and N-acetyl-lactosamine. Human antisera were used to identify A and B antigens. In all groups N-acetyl-lactosamine and H antigen were found on the cell membranes of the spinous cell layer. N-acetyl-lactosamine was present mainly...

  14. Membrane proteomics characterization of brush border membrane proteins of mice intestinal mucosa : case study: cholesterol absorption

    OpenAIRE

    Tsirogianni, Eirini

    2009-01-01

    The epithelial absorbing cells of the small intestinal villi, the enterocytes, are the main protagonists for the transport of nutrients from the intestinal lumen to the interstitial fluids. The oriented flow of nutrients is carried out by different and complementary transport systems present in the apical and the basolateral domains of the enterocyte’s plasma membrane. One of the distinctive characteristics of those intestinal cells is the presence of numerous structurally distinct protrusion...

  15. Calcium absorption by Cav1.3 induces terminal web myosin II phosphorylation and apical GLUT2 insertion in rat intestine.

    Science.gov (United States)

    Mace, Oliver J; Morgan, Emma L; Affleck, Julie A; Lister, Norma; Kellett, George L

    2007-04-15

    Glucose absorption in rat jejunum involves Ca(2+)- and PKC betaII-dependent insertion of GLUT2 into the apical membrane. Ca(2+)-induced rearrangement of the enterocyte cytoskeleton is thought to enhance paracellular flow. We have therefore investigated the relationships between myosin II regulatory light chain phosphorylation (RLC(20)), absorption of glucose, water and calcium, and mannitol clearance. ML-7, an inhibitor of myosin light chain kinase, diminished the phloretin-sensitive apical GLUT2 but not the phloretin-insensitive SGLT1 component of glucose absorption in rat jejunum perfused with 75 mM glucose. Western blotting and immunocytochemistry revealed marked decreases in RLC(20) phosphorylation in the terminal web and in the levels of apical GLUT2 and PKC betaII, but not SGLT1. Perfusion with phloridzin or 75 mM mannitol, removal of luminal Ca(2+), or inhibition of unidirectional (45)Ca(2+) absorption by nifedipine exerted similar effects. ML-7 had no effect on the absorption of 10 mM Ca(2+), nor clearance of [(14)C]-mannitol, which was less than 0.7% of the rate of glucose absorption. Water absorption did not correlate with (45)Ca(2+) absorption or mannitol clearance. We conclude that the Ca(2+) necessary for contraction of myosin II in the terminal web enters via an L-type channel, most likely Ca(v)1.3, and is dependent on SGLT1. Moreover, terminal web RLC(20) phosphorylation is necessary for apical GLUT2 insertion. The data confirm that glucose absorption by paracellular flow is negligible, and show further that paracellular flow makes no more than a minimal contribution to jejunal Ca(2+) absorption at luminal concentrations prevailing after a meal.

  16. Detection of peste des petits ruminants virus antigen using immunofiltration and antigen-competition ELISA methods.

    Science.gov (United States)

    Raj, G Dhinakar; Rajanathan, T M C; Kumar, C Senthil; Ramathilagam, G; Hiremath, Geetha; Shaila, M S

    2008-06-22

    Peste des petits ruminants (PPR) is one of the most economically important diseases affecting sheep and goats in India. An immunofiltration-based test has been developed using either mono-specific serum/monoclonal antibodies (mAb) prepared against a recombinant truncated nucleocapsid protein of rinderpest virus (RPV) cross-reactive with PPR virus. This method consists of coating ocular swab eluate from suspected animals onto a nitrocellulose membrane housed in a plastic module, which is allowed to react with suitable dilutions of a mAb or a mono-specific polyclonal antibody. The antigen-antibody complex formed on the membrane is then detected by protein A-colloidal gold conjugate, which forms a pink colour. In the immunofiltration test, concordant results were obtained using either PPRV mAb or mono-specific serum. Another test, an antigen-competition ELISA which relies on the competition between plate-coated recombinant truncated 'N' protein of RPV and the PPRV 'N' protein present in ocular swab eluates (sample) for binding to the mono-specific antibody against N protein of RPV (in liquid phase) was developed. The cut-off value for this test was established using reverse transcription polymerase chain reaction (RT-PCR) positive and negative oculo-nasal swab samples. Linear correlation between percent inhibition (PI) values in antigen-competition ELISA and virus infectivity titres was 0.992. Comparison of the immunofiltration test with the antigen-competition ELISA yielded a sensitivity of 80% and specificity of 100%. These two tests can serve as a screening (immunofiltration) and confirmatory (antigen-competition ELISA) test, respectively, in the diagnosis of PPR in sheep or goats. PMID:18182256

  17. The apical complex provides a regulated gateway for secretion of invasion factors in Toxoplasma.

    Directory of Open Access Journals (Sweden)

    Nicholas J Katris

    2014-04-01

    Full Text Available The apical complex is the definitive cell structure of phylum Apicomplexa, and is the focus of the events of host cell penetration and the establishment of intracellular parasitism. Despite the importance of this structure, its molecular composition is relatively poorly known and few studies have experimentally tested its functions. We have characterized a novel Toxoplasma gondii protein, RNG2, that is located at the apical polar ring--the common structural element of apical complexes. During cell division, RNG2 is first recruited to centrosomes immediately after their duplication, confirming that assembly of the new apical complex commences as one of the earliest events of cell replication. RNG2 subsequently forms a ring, with the carboxy- and amino-termini anchored to the apical polar ring and mobile conoid, respectively, linking these two structures. Super-resolution microscopy resolves these two termini, and reveals that RNG2 orientation flips during invasion when the conoid is extruded. Inducible knockdown of RNG2 strongly inhibits host cell invasion. Consistent with this, secretion of micronemes is prevented in the absence of RNG2. This block, however, can be fully or partially overcome by exogenous stimulation of calcium or cGMP signaling pathways, respectively, implicating the apical complex directly in these signaling events. RNG2 demonstrates for the first time a role for the apical complex in controlling secretion of invasion factors in this important group of parasites.

  18. Evaluation of the distortion rate of panoramic and peri apical radiographs in erupted third molar inclination

    International Nuclear Information System (INIS)

    Panoramic and peri apical radiographs are normally used in impacted third molar teeth surgeries. The aim of the present study was to evaluate and compare the distortion of the erupted third molar teeth on panoramic and peri apical radiographs. Patients and Methods: A total of 44 radiographs were obtained of 22 patients (age range, 18-24 years) referred to the faculty of dentistry for orthodontic treatment. A plaster cast was prepared and panoramic radiography was taken for all patients to plan the orthodontic treatment and peri apical radiography was taken for investigation of tooth structure details. Therefore, a total of 66 views and samples were studied by two methods: 1) Measuring the angle between the longitudinal plane of the third molar and occlusal plane. 2) Measuring the angle between the longitudinal plane of second and third molar. Finally, 132 records were evaluated by one individual. Results: There was no significant statistical difference between the mean position of the third molar on panoramic, peri apical radiographs and the casts. However, measurements of the third molars on peri apical radiographs were slightly closer to the measurements of the casts compared to the panoramic radiographs. Conclusion: Distortion does not have a specific effect on the diagnosis of the position of the third erupted molars by peri apical or panoramic radiographs, though various studies have shown that these radiographs have an amount of distortion and peri apical radiographical distortion is less than that in panoramic radiography.

  19. Apical constriction initiates new bud formation during monopodial branching of the embryonic chicken lung

    Science.gov (United States)

    Kim, Hye Young; Varner, Victor D.; Nelson, Celeste M.

    2013-01-01

    Branching morphogenesis sculpts the airway epithelium of the lung into a tree-like structure to conduct air and promote gas exchange after birth. In the avian lung, a series of buds emerges from the dorsal surface of the primary bronchus via monopodial branching to form the conducting airways; anatomically, these buds are similar to those formed by domain branching in the mammalian lung. Here, we show that monopodial branching is initiated by apical constriction of the airway epithelium, and not by differential cell proliferation, using computational modeling and quantitative imaging of embryonic chicken lung explants. Both filamentous actin and phosphorylated myosin light chain were enriched at the apical surface of the airway epithelium during monopodial branching. Consistently, inhibiting actomyosin contractility prevented apical constriction and blocked branch initiation. Although cell proliferation was enhanced along the dorsal and ventral aspects of the primary bronchus, especially before branch formation, inhibiting proliferation had no effect on the initiation of branches. To test whether the physical forces from apical constriction alone are sufficient to drive the formation of new buds, we constructed a nonlinear, three-dimensional finite element model of the airway epithelium and used it to simulate apical constriction and proliferation in the primary bronchus. Our results suggest that, consistent with the experimental results, apical constriction is sufficient to drive the early stages of monopodial branching whereas cell proliferation is dispensable. We propose that initial folding of the airway epithelium is driven primarily by apical constriction during monopodial branching of the avian lung. PMID:23824575

  20. Variability interexaminer of chronic apical periodontitis diagnostics in panoramic radiographs

    International Nuclear Information System (INIS)

    The accuracy of radiological diagnosis of Apical Periodontitis (AP) is reported between examiners with differents levels of clinical experience using panoramic radiographs. 1032 teeth in 41 panoramic radiographs have been diagnosed. The evaluation of the x-rays and radiological diagnosis is performed by three independent dentists for AP to each tooth. The teeth have presented without radiological signs of AP in 96.6%. The number of teeth classified with an uncertain diagnosis of AP at the has been inversely proportional to the number of years of experience of the examiner. The examiner A has been used as a reference and compared the diagnoses of other examiners.The percentage agreement for all teeth was 95% for examiner B and 94% for examiner C. Kappa for all has been of 0.44 for examiner B and 0.43 for examiner C. The different categories were analyzed separately, the inferior teeth and the anterior teeth have obtained the best results. The level of clinical experience of the examiner has been indifferent in the diagnosis of AP in panoramic radiographs; however, it is important experience in the number of radiographs extras that will be needed to give a radiological diagnosis definitive. The panoramic radiographs were reliable diagnostic tools for AP. (author)

  1. Emergence of an Apical Epithelial Cell Surface In Vivo.

    Science.gov (United States)

    Sedzinski, Jakub; Hannezo, Edouard; Tu, Fan; Biro, Maté; Wallingford, John B

    2016-01-11

    Epithelial sheets are crucial components of all metazoan animals, enclosing organs and protecting the animal from its environment. Epithelial homeostasis poses unique challenges, as addition of new cells and loss of old cells must be achieved without disrupting the fluid-tight barrier and apicobasal polarity of the epithelium. Several studies have identified cell biological mechanisms underlying extrusion of cells from epithelia, but far less is known of the converse mechanism by which new cells are added. Here, we combine molecular, pharmacological, and laser-dissection experiments with theoretical modeling to characterize forces driving emergence of an apical surface as single nascent cells are added to a vertebrate epithelium in vivo. We find that this process involves the interplay between cell-autonomous actin-generated pushing forces in the emerging cell and mechanical properties of neighboring cells. Our findings define the forces driving this cell behavior, contributing to a more comprehensive understanding of epithelial homeostasis. PMID:26766441

  2. Obturating teeth with wide open apices using mineral trioxide aggregate: a case report.

    Science.gov (United States)

    Levenstein, H

    2002-07-01

    The conventional approach in handling a tooth with a wide open apex requiring endodontic treatment is by means of a procedure called apexification. The objective of treatment is to introduce calcium hydroxide mixed with sterile water or local anaesthetic into the root canal to create a hard-tissue-like formation or an apical plug to prevent extrusion of filling materials during obturation of teeth with wide open apices. This procedure may take anything from 6 months to 2 years. In 1999 a new material called mineral trioxide aggregate (MTA) was introduced to the dental profession for clinical use which has the ability to create an apical plug within a few weeks.

  3. Murine antigen-induced arthritis.

    NARCIS (Netherlands)

    Berg, W.B. van den; Joosten, L.A.B.; Lent, P.L.E.M. van

    2007-01-01

    Antigen induced arthritis is a unilateral T-cell driven model caused by direct injection of an antigen into the knee joint of a FCA preimmunized animal. The chronicity is determined by antigen retention in avascular structures of the joint through charge mediated binding or antibody mediated trappin

  4. Membrane fusion

    DEFF Research Database (Denmark)

    Bendix, Pól Martin

    2015-01-01

    At Stanford University, Boxer lab, I worked on membrane fusion of small unilamellar lipid vesicles to flat membranes tethered to glass surfaces. This geometry closely resembles biological systems in which liposomes fuse to plasma membranes. The fusion mechanism was studied using DNA zippering...... between complementary strands linked to the two apposing membranes closely mimicking the zippering mechanism of SNARE fusion complexes....

  5. Mint essential oil can induce or inhibit potato sprouting by differential alteration of apical meristem.

    Science.gov (United States)

    Teper-Bamnolker, Paula; Dudai, Nativ; Fischer, Ravit; Belausov, Eduard; Zemach, Hanita; Shoseyov, Oded; Eshel, Dani

    2010-06-01

    Sprouting of potatoes during storage, due to tuber dormancy release, is associated with weight loss and softening. Sprout-preventing chemicals, such as chlorpropham (CIPC), can negatively impact the environment and human health. Monthly thermal fogging with mint (Mentha spicata L.) essential oil (MEO) inhibited sprouting in eight potato cultivars during large-volume 6-month storage: the tubers remained firm with 38% lower weight loss after 140 days of storage. The sprout-inhibitory action may be nullified: treated tubers washed with water resumed sprouting within days, with reduced apical dominance. MEO application caused local necrosis of the bud meristem, and a few weeks later, axillary bud (AX) growth was induced in the same sprouting eye. MEO components analysis showed that 73% of its content is the monoterpene R-carvone. Tubers treated with synthetic R-carvone in equivalent dose, 4.5 microl l(-1), showed an inhibitory effect similar to that of MEO. Surprisingly, 0.5 microl l(-1) of MEO or synthetic R-carvone catalyzed AX sprouting in the tuber. To the best of our knowledge, this is the first report of an essential oil vapor inducing early sprouting of potato tubers. R-carvone caused visible damage to the meristem membrane at sprout-inhibiting, but not sprout-inducing doses, suggesting different underlying mechanisms. After 5 days' exposure to R-carvone, its derivatives transcarveol and neo-dihydrocarveol were found in buds of tubers treated with the inhibitory dose, suggesting biodegradation. These experiments demonstrate the potential of MEO vapor as an environmentally friendly alternative to CIPC in stored potatoes and as a research tool for the control of sprouting in plants. PMID:20390295

  6. Mint essential oil can induce or inhibit potato sprouting by differential alteration of apical meristem.

    Science.gov (United States)

    Teper-Bamnolker, Paula; Dudai, Nativ; Fischer, Ravit; Belausov, Eduard; Zemach, Hanita; Shoseyov, Oded; Eshel, Dani

    2010-06-01

    Sprouting of potatoes during storage, due to tuber dormancy release, is associated with weight loss and softening. Sprout-preventing chemicals, such as chlorpropham (CIPC), can negatively impact the environment and human health. Monthly thermal fogging with mint (Mentha spicata L.) essential oil (MEO) inhibited sprouting in eight potato cultivars during large-volume 6-month storage: the tubers remained firm with 38% lower weight loss after 140 days of storage. The sprout-inhibitory action may be nullified: treated tubers washed with water resumed sprouting within days, with reduced apical dominance. MEO application caused local necrosis of the bud meristem, and a few weeks later, axillary bud (AX) growth was induced in the same sprouting eye. MEO components analysis showed that 73% of its content is the monoterpene R-carvone. Tubers treated with synthetic R-carvone in equivalent dose, 4.5 microl l(-1), showed an inhibitory effect similar to that of MEO. Surprisingly, 0.5 microl l(-1) of MEO or synthetic R-carvone catalyzed AX sprouting in the tuber. To the best of our knowledge, this is the first report of an essential oil vapor inducing early sprouting of potato tubers. R-carvone caused visible damage to the meristem membrane at sprout-inhibiting, but not sprout-inducing doses, suggesting different underlying mechanisms. After 5 days' exposure to R-carvone, its derivatives transcarveol and neo-dihydrocarveol were found in buds of tubers treated with the inhibitory dose, suggesting biodegradation. These experiments demonstrate the potential of MEO vapor as an environmentally friendly alternative to CIPC in stored potatoes and as a research tool for the control of sprouting in plants.

  7. AMP-activated protein kinase inhibits alkaline pH- and PKA-induced apical vacuolar H+-ATPase accumulation in epididymal clear cells.

    Science.gov (United States)

    Hallows, Kenneth R; Alzamora, Rodrigo; Li, Hui; Gong, Fan; Smolak, Christy; Neumann, Dietbert; Pastor-Soler, Núria M

    2009-04-01

    Acidic luminal pH and low [HCO(3)(-)] maintain sperm quiescent during maturation in the epididymis. The vacuolar H(+)-ATPase (V-ATPase) in clear cells is a major contributor to epididymal luminal acidification. We have shown previously that protein kinase A (PKA), acting downstream of soluble adenylyl cyclase stimulation by alkaline luminal pH or HCO(3)(-), induces V-ATPase apical membrane accumulation in clear cells. Here we examined whether the metabolic sensor AMP-activated protein kinase (AMPK) regulates this PKA-induced V-ATPase apical membrane accumulation. Immunofluorescence labeling of rat and non-human primate epididymides revealed specific AMPK expression in epithelial cells. Immunofluorescence labeling of rat epididymis showed that perfusion in vivo with the AMPK activators 5-aminoimidazole-4-carboxamide-1-beta-d-ribofuranoside (AICAR) or A-769662 induced a redistribution of the V-ATPase into subapical vesicles, even in the presence of a luminal alkaline (pH 7.8) buffer compared with that of controls perfused without drug. Moreover, preperfusion with AICAR blocked the PKA-mediated V-ATPase translocation to clear cell apical membranes induced by N(6)-monobutyryl-cAMP (6-MB-cAMP). Purified PKA and AMPK both phosphorylated V-ATPase A subunit in vitro. In HEK-293 cells [(32)P]orthophosphate in vivo labeling of the A subunit increased following PKA stimulation and decreased following RNA interference-mediated knockdown of AMPK. Finally, the extent of PKA-dependent in vivo phosphorylation of the A subunit increased with AMPK knockdown. In summary, our findings suggest that AMPK inhibits PKA-mediated V-ATPase apical accumulation in epididymal clear cells, that both kinases directly phosphorylate the V-ATPase A subunit in vitro and in vivo, and that AMPK inhibits PKA-dependent phosphorylation of this subunit. V-ATPase activity may be coupled to the sensing of acid-base status via PKA and to metabolic status via AMPK. PMID:19211918

  8. Frecuencia de periodontitis apical en tratamientos endodónticos de pregrado Frequency of apical periodontitis in endodontic treatment in undergraduate

    OpenAIRE

    León, P; MJ Ilabaca; M Alcota; FE González

    2011-01-01

    La periodontitis apical es una enfermedad de los tejidos periapicales de etiología bacteriana. Su tratamiento requiere erradicar los microorganismos del conducto radicular y obturarlo para lograr la reparación posterior. El objetivo de éste trabajo fue determinar la frecuencia de periodontitis apical en la población que asiste a la clínica de Endodoncia de la Facultad de Odontología de la Universidad de Chile y su relación con diferentes variables: diagnóstico específico, edad, sexo, agudas v...

  9. Pulp microbiology of complete teeth with idiopathic apical lesions.

    Directory of Open Access Journals (Sweden)

    Patricia Rodríguez

    2009-11-01

    Full Text Available Introduction: Periapical changes named as lesions, in teeth with full crown integrity and without history of trauma, do not show a clear aetiology. Objective: To determine the presence of microorganisms in pulp dental tissue will clarify the cause of its death and therefore the damage to periodontal tissues. Materials and methods: From people between 10 and 39 years old, 23 teeth were selected. The samples were taken with paper points and 0.8 sterile files, and were transported in VMGA III medium, to be processed in the following 24 hours after they were taken and sowed in Brucella-agar. Results: The most affected teeth were upper central incisors, 43.8%. From the 23 studied teeth, microbiological grow was seen on 20 teeth. The following microorganisms species were identified: Fusobacterium spp., 25%, Eubacterium spp., 15%; Peptostreptococcus spp., 10%; Campylobacter spp., 10%; gram negative enteric bacteria, 10%; Porphyromonas gingivalis, 10%; Prevotella intermedia, 5%; Eikenellia corrodens, 5%; Dialister pneumosintes, 5%; and yeasts, 5%. There was no growing evidence of Actinomyces actinomycetemcomitans, Tanerella forsythensis and Streptococcus β  hemolytic. Discussion and conclusions: Sound pulp dental tissue is sterile; an injury over it will cause its inflammation, degeneration, death and bacterial contamination. Results in the present study clearly show the presence of microorganisms in closed apical dental lesions of endodontic origin. In same manner, it was seen that a great part of microorganisms species found can be regarded as periodontal pathogens. This could suggest a management with an endodontic, a periodontic and a pharmacological combined treatment.

  10. Advances in molecular imaging of prostate cancer targeting prostate specific membrane antigen with small molecular radionuclide probes%以核素标记前列腺特异性膜抗原小分子抑制剂为探针的前列腺癌分子影像研究进展

    Institute of Scientific and Technical Information of China (English)

    胡四龙(综述); 许晓平; 章英剑(审校)

    2015-01-01

    Prostate speciifc membrane antigen (PSMA) is expressed in normal prostate epithelial cells and strongly up regulated in initial and advanced prostate carcinoma. Therefore, PSMA is an attractive molecular target for the detection of primary and metastatic lesions. It plays an important role in the diagnosis, staging, prognostic evaluation and targeted treatment in prostate carcinoma patients. This review will summarize the progress of small molecular radionuclide probes, which targeted PSMA, for visualizing prostate cancer.%前列腺特异性膜抗原(PSMA)高表达于前列腺癌及其转移灶中,可作为生物分子靶点,在前列腺癌的早期诊断、分期、复发检测、预后判断及靶向治疗中具有重要价值。本文重点综述以核素标记PSMA小分子抑制剂为探针的前列腺癌分子影像研究进展。

  11. Magnetic Resonance Imaging of Transient Left Ventricular Apical Ballooning Related to Emotional Stress: a Case Report

    International Nuclear Information System (INIS)

    Transient left ventricular apical ballooning is characterized by transient wall motion abnormalities involving the left ventricular apex and mid-ventricle in the absence of coronary arterial occlusion. A 66-year-old woman presented to the emergency department with chest pain that mimicked acute myocardial infarction. An aortogram showed akinesis from the mid to apical left ventricle with sparing of the basal segments. Four days later, she underwent MRI, which demonstrated characteristic apical contractile dysfunction, the same as the aortogram, without evidence of myocardial infarction on the MRI. Two weeks later, her symptoms were resolved and follow-up echocardiography showed normal ventricular function. We suggest that MRI might be an integrated imaging diagnostic tool for the diagnosis of this syndrome, which demonstrated characteristic apical contractile dysfunction with performing cine MRI, the absence of significant coronary artery stenosis with performing coronary MR angiography and the absence of myocardial infarction with performing contrast enhanced delayed MRI

  12. Bacterial Outer Membrane Vesicles and Vaccine Applications

    OpenAIRE

    Acevedo, Reinaldo; Fernández, Sonsire; Zayas, Caridad; Acosta, Armando; Sarmiento, Maria Elena; Valerie A. Ferro; Rosenqvist, Einar; Campa, Concepcion; Cardoso, Daniel; Garcia, Luis; Perez, Jose Luis

    2014-01-01

    Vaccines based on outer membrane vesicles (OMV) were developed more than 20 years ago against Neisseria meningitidis serogroup B. These nano-sized structures exhibit remarkable potential for immunomodulation of immune responses and delivery of meningococcal antigens or unrelated antigens incorporated into the vesicle structure. This paper reviews different applications in OMV Research and Development (R&D) and provides examples of OMV developed and evaluated at the Finlay Institute in Cuba. A...

  13. BACTERIAL OUTER MEMBRANE VESICLES AND VACCINE APPLICATIONS

    OpenAIRE

    Reinaldo eAcevedo; Sonsire eFernandez; Caridad eZayas; Armando eAcosta; Maria Elena Sarmiento; Valerie A. Ferro; Einar eRosenqvist; Concepcion eCampa; Daniel eCardoso; Luis eGarcia; Jose Luis Perez

    2014-01-01

    Vaccines based on outer membrane vesicles (OMV) were developed more than 20 years ago against Neisseria meningitidis serogroup B. These nano-sized structures exhibit remarkable potential for immunomodulation of immune responses and delivery of self meningococcal antigens or unrelated antigens incorporated into the vesicle structure. This paper reviews different applications in OMV Research and Development (R&D) and provides examples of OMV developed and evaluated at the Finlay Institute in Cu...

  14. Effect of colchicine on rat small intestinal absorptive cells. II. Distribution of label after incorporation of [3H]fucose into plasma membrane glycoproteins

    International Nuclear Information System (INIS)

    By means of radioautography the influence was tested of various periods (5, 15, 30, 40 min, 2 hr) of pretreatment with colchicine, administered intraperitoneally to rats at a dosage of 0.5 mg/100 g of body weight, on the intracellular pathway of [3H]fucose in absorptive cells of the small intestine. Administration of colchicine for 30 min and longer time intervals causes delay in the insertion of [3H]fucose into the oligosaccharide chains of glycoconjugates in the Golgi apparatus, and results in redistribution of the label apparent over the different portions of the plasma membrane. In controls, at 2 and 4 hr after administration of [3H]fucose the apical plasma membrane is strongly labeled. Colchicine causes equalization of the reaction of apical and basolateral regions of the plasma membrane: the number of silver grains attributable to the apical plasma membrane is reduced; following treatment with colchicine, apical portions of the plasma membrane comprise 31.6 +/- 1.8% of the silver grains, 38.6 +/- 3.8% are attributable to basolateral membrane regions. The colchicine-induced equalization of the density of label of apical and basolateral regions of the plasma membrane, in addition to the occurrence of basolateral microvillus borders, suggests microtubules to be important in the maintenance of the polar organization of small intestinal absorptive cells

  15. Effect of needle insertion depth and apical diameter on irrigant extrusion in simulated immature permanent teeth

    Directory of Open Access Journals (Sweden)

    AKSEL Hacer

    2014-09-01

    Full Text Available The aim of this study was to compare the amount of irrigant extrusion in simulated immature permanent teeth when the apical diameter and needle insertion depth were varied. Thirty single-rooted maxillary incisors with straight root canals were selected. The root length was standardized to a length of 9 mm. The teeth were divided into two experimental groups according to the degree of apical enlargement (n = 15. The apices were enlarged to a diameter of 1.10 mm or 1.70 mm by using a #3 or #6 peeso reamer, respectively, to simulate immature teeth. The irrigation solution was applied 2 or 4 mm short of the working length (WL in each experimental group. The glass vial model was used for the collection of extruded irrigant beyond the root apex. A two-way repeated measures analysis of variance test showed that there was no significant difference between different needle insertion depths (2 and 4 mm short of the WL in the group with an apical diameter of 1.70 mm (p > 0.05. In the group with an apical diameter of 1.10 mm, a 32% increase in irrigant extrusion was observed when the needle was positioned at 2 mm (p < 0.05. Regarding the effect of apical diameter, the group with a diameter of 1.70 mm showed more apical extrusion of the irrigant (34% increase for the needle positioned at 2 mm and 68% increase for the needle positioned at 4 mm. It was observed that the needle insertion depth and apical diameter have a significant effect on irrigant extrusion in immature permanent teeth.

  16. Comparison of Apical Axial Derotation between Adolescent Idiopathic and Neuromuscular Scoliosis with Pedicle Screw Instrumentation

    OpenAIRE

    Modi, Hitesh N.; Suh, Seung-Woo; Srinivasalu, S.; Mehta, Satyen; Yang, Jae-Hyuk

    2008-01-01

    Study Design A retrospective study. Purpose To compare outcomes of apical derotation with pedicle screws in idiopathic and neuromuscular scoliosis (NMS). Overview of Literature No information about apical derotation in NMS with pedicle screws is available. Methods We performed deformity correcting surgery using pedicle screw constructs on 12 adolescent idiopathic scoliosis (AIS) patients (mean age 14.1 years) and 16 NMS patients (mean age 16.5 years). Preoperative, postoperative, and final fo...

  17. Apical ballooning with mid-ventricular obstruction: the many faces of Takotsubo cardiomyopathy

    OpenAIRE

    Spadotto, Veronica; ElMaghawry, Mohamed; Zorzi, Alessandro; Migliore, Federico; Marra, Martina Perazzolo

    2013-01-01

    Takotsubo cardiomyopathy (TTC) is a transient left ventricular dysfunction due to akinesia of the left-ventricular (LV) mid-apical segments (apical ballooning), which can cause severe reduction in LV systolic function. The typical clinical picture of TTC include chest pain, electrocardiographic changes consisting of mild ST-segment elevation followed by diffuse deep T-wave inversion, QTc interval prolongation and mild troponin release in the absence of significant coronary stenoses. The syndr...

  18. 24 Hour ST Segment Analysis in Transient Left Ventricular Apical Ballooning

    OpenAIRE

    Frank Bode; Christof Burgdorf; Heribert Schunkert; Volkhard Kurowski

    2013-01-01

    OBJECTIVE: The etiologic basis of transient left ventricular apical ballooning, a novel cardiac syndrome, is not clear. Among the proposed pathomechanisms is coronary vasospasm. Long-term ST segment analysis may detect vasospastic episodes but has not been reported. METHODS: 30 consecutive patients with transient left ventricular apical ballooning, left ventricular dysfunction and normal or near-normal coronary arteries were investigated. A 24-hour Holter ECG was obtained after emergency admi...

  19. RNG1 is a Late Marker of the Apical Polar Ring in Toxoplasma gondii

    OpenAIRE

    Tran, Johnson Q.; de Leon, Jessica C.; Li, Catherine; Huynh, My-Hang; Beatty, Wandy; Morrissette, Naomi S.

    2010-01-01

    The asexually proliferating stages of apicomplexan parasites cause acute symptoms of diseases such as malaria, cryptosporidiosis and toxoplasmosis. These stages are characterized by the presence of two independent microtubule organizing centers (MTOCs). Centrioles are found at the poles of the intranuclear spindle. The apical polar ring (APR), a MTOC unique to apicomplexans, organizes subpellicular microtubules which impose cell shape and apical polarity on these protozoa. Here we describe th...

  20. Frecuencia de periodontitis apical en tratamientos endodónticos de pregrado Frequency of apical periodontitis in endodontic treatment in undergraduate

    Directory of Open Access Journals (Sweden)

    P León

    2011-12-01

    Full Text Available La periodontitis apical es una enfermedad de los tejidos periapicales de etiología bacteriana. Su tratamiento requiere erradicar los microorganismos del conducto radicular y obturarlo para lograr la reparación posterior. El objetivo de éste trabajo fue determinar la frecuencia de periodontitis apical en la población que asiste a la clínica de Endodoncia de la Facultad de Odontología de la Universidad de Chile y su relación con diferentes variables: diagnóstico específico, edad, sexo, agudas vs. crónicas, éxito/fracaso del tratamiento y rehabilitación post-tratamiento. Se recolectó información de 292 dientes de pacientes tratados en dicha clínica, analizándose la información de aquellos con dientes diagnosticados con algún tipo de periodontitis apical clasificándolo según la nueva nomenclatura de la asociación de endodoncia americana. La frecuencia de periodontitis apical fue de un 36.7% (92 pacientes, de los cuales un 77.8% presentaron patologías de tipo crónicas. Un 75% de los pacientes pertenecieron al sexo femenino y la 5ta década representó un 34.8% de los pacientes. El 100% de los pacientes que acudieron a control presentó éxito en sus tratamientos según variables clínico-radiográficas, mientras que la restauración más frecuente post-tratamiento fue la resina compuesta (52.4%. Dado el pequeño número de pacientes que concurrieron a la citación de control para determinar el éxito del tratamiento, esta variable debe ser considerada como preliminar. Nuestros resultados mostraron que la condición mas frecuente fue la periodontitis apical de tipo crónica, afectando más a mujeres y a la 5ta década.Apical periodontitis is a microbially induced inflammatory disease of the periapical tissues. Its treatment requires eliminating microorganism from root canal and sealing it properly to induce further tissue repair. The aim of this study was to determine the frequency of apical periodontitis and its distribution by

  1. Comparison of Irrigation Penetration into the Apical Part of Canals in Hand and Rotary Instrumentations

    Directory of Open Access Journals (Sweden)

    M. Khabiri

    2007-01-01

    Full Text Available Introduction: The penetration of irrigating solution to the apical one third of canals and removal of debris are dependent on the final size of the instruments and instrumentation techniques used in the canals. The purpose of this study was to evaluate the effect of final instrument size, on irrigation penetration into the apical part of canals in hand K-file instrumentation versus rotary system of Hero 642.Methods and Materials: The mesiobuccal canals of 48 first mandibular molar teeth were selected for this study. The teeth were divided into 2 groups of 24 in each and the mesiobuccal canals were instrumented by hand K-file or rotary system of Hero 642 at 2 stages. After each stage, a contrast medium was injected into the canals and radiographs were taken by RVG system. The irrigation penetration was measured in radiographs by Diamax software. The data were analyzed using t – student test.Results: This study showed that instrumentation up to # 25 file is not enough for irrigation penetration into the apical area. Also by more flaring the canals, more irrigating solution penetrates into the apical part of canals (P 0 0.001, but the difference between hand and rotary systems was not statistically significant (P > 0.05.Discussion: According to this study, instrumentation up to # 30 file results in better irrigation penetration into the apical area. The flaring of the canals is essential for better cleaning and irrigation of apical area.

  2. Calcium-enriched mixture cement as artificial apical barrier: A case series

    Directory of Open Access Journals (Sweden)

    Ali Nosrat

    2011-01-01

    Full Text Available In comparison to the conventional apexification using calcium hydroxide, artificial apical barrier technique is more valuable and less time consuming. This article describes successful use of calcium-enriched mixture (CEM cement as an artificial apical barrier in open apices. In this study, 13 single-rooted teeth with necrotic pulps and open apices were treated non-surgically. After copious irrigation of the root canals with NaOCl 5.25% and gentle filing, based on need for interappointment dressing, treatments were followed by CEM cement (BioniqueDent, Tehran, Iran apical plug insertion in the first or second appointment. All cases were then permanently restored. All subjects were followed until radiographic evidence of periradicular healing was seen (mean 14.5 months. Clinically, all cases were functional and asymptomatic and complete osseous healing was observed in all the teeth. Considering the biological properties of CEM cement, this new endodontic biomaterial might be appropriate to be used as artificial apical barrier in the open apex teeth.

  3. Transposon-mediated alteration of TaMATE1B expression in wheat confers constitutive citrate efflux from root apices.

    Science.gov (United States)

    Tovkach, Andriy; Ryan, Peter R; Richardson, Alan E; Lewis, David C; Rathjen, Tina M; Ramesh, Sunita; Tyerman, Stephen D; Delhaize, Emmanuel

    2013-02-01

    The TaMATE1B gene (for multidrug and toxic compound extrusion) from wheat (Triticum aestivum) was isolated and shown to encode a citrate transporter that is located on the plasma membrane. TaMATE1B expression in roots was induced by iron deficiency but not by phosphorus deficiency or aluminum treatment. The coding region of TaMATE1B was identical in a genotype showing citrate efflux from root apices (cv Carazinho) to one that lacked citrate efflux (cv Egret). However, sequence upstream of the coding region differed between these two genotypes in two ways. The first difference was a single-nucleotide polymorphism located approximately 2 kb upstream from the start codon in cv Egret. The second difference was an 11.1-kb transposon-like element located 25 bp upstream of the start codon in cv Carazinho that was absent from cv Egret. The influence of these polymorphisms on TaMATE1B expression was investigated using fusions to green fluorescent protein expressed in transgenic lines of rice (Oryza sativa). Fluorescence measurements in roots of rice indicated that 1.5- and 2.3-kb regions upstream of TaMATE1B in cv Carazinho (which incorporated 3' regions of the transposon-like element) generated 20-fold greater expression in the apical 1 mm of root compared with the native promoter in cv Egret. By contrast, fluorescence in more mature tissues was similar in both cultivars. The presence of the single-nucleotide polymorphism alone consistently generated 2-fold greater fluorescence than the cv Egret promoter. We conclude that the transposon-like element in cv Carazinho extends TaMATE1B expression to the root apex, where it confers citrate efflux and enhanced aluminum tolerance.

  4. Regulation of antigen presentation by acidic pH

    OpenAIRE

    1990-01-01

    The effect of pH on functional association of peptide antigens with APC membranes was investigated by using aldehyde-fixed B cells and class II- restricted T cell hybridomas to assess antigen/MHC complex formation. The results indicated that the rate and extent of functional peptide binding was markedly increased at pH 5.0 as compared with pH 7.3. The pH dependence of binding was preserved after pretreatment of fixed APC with pH 5.0 buffer, suggesting that pH had a direct effect on the intera...

  5. Identification of a membrane-associated interleukin 1 in macrophages.

    OpenAIRE

    Kurt-Jones, E A; Beller, D I; Mizel, S B; Unanue, E R

    1985-01-01

    We have found a surface membrane-associated interleukin 1 (IL-1) with potent thymocyte and T-cell stimulatory activity on peptone-elicited peritoneal macrophages. The IL-1 activity was demonstrated on both fixed macrophage monolayers and on isolated membranes from unfixed macrophages. Membrane IL-1 was induced by adherence and/or by adding heat-killed Listeria monocytogenes to macrophage cultures. The macrophage membrane IL-1 was similar functionally and antigenically to soluble IL-1, but its...

  6. Detection of serum anti-melanocyte antibodies and identification of related antigens in patients with vitiligo.

    Science.gov (United States)

    Zhu, M C; Liu, C G; Wang, D X; Zhan, Z

    2015-12-07

    We detected autoantibodies against melanocytes in serum samples obtained from 50 patients, including 4 with HBV, with vitiligo and identified the associated membrane antigens. Heat shock protein 70 (HSP70) and anti-tyrosinase-related protein 1 (TRP-1) antibody levels were analyzed. The associated antigens in normal human melanocyte were identified by immunofluorescence. Autoantibodies against melanocyte membrane and cytoplasmic proteins were detected by western blot. Membrane antigens with higher frequencies were identified by protein mass spectrometry. The HSP70 and anti-TRP-1 antibody levels (N = 70; 10 with HBV) were detected by ELISA. The specific antigens were detected in melanocyte cytoplasm and membrane (40/50; 80% incidence; western blot). The autoantibodies reacted with several membrane antigens with approximate molecular weights (Mr) of 86,000, 75,000, 60,000, 52,000, and 44,000 (strip positive rates: 36, 58, 22, 2, and 2%, respectively). Thirty percent of the patients showed the presence of cytoplasmic antigens (Mr: 110,000, 90,000, 75,000, 50,000, and 400,000; strip positive rates: 12, 4, 12, 10, and 2%, respectively). Fifteen and 5% of the healthy subjects showed positive expression of membrane and cytoplasmic antigens, respectively. Protein mass spectrometry predicted membrane proteins with Mr of 86,000 and 75,000 and 60,000 to be Lamin A /C and Vimentin X1, respective. High titers of anti-TRP-1 antibody were detected and showed positive correlation with HSP70 (r = 0. 927, P vitiligo, which might assist future investigations into autoimmune pathogenesis of vitiligo and formation of autoantibodies. HBV infection was correlated to vitiligo.

  7. Light Responsive Polymer Membranes: A Review

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    Fiore Pasquale Nicoletta

    2012-03-01

    Full Text Available In recent years, stimuli responsive materials have gained significant attention in membrane separation processes due to their ability to change specific properties in response to small external stimuli, such as light, pH, temperature, ionic strength, pressure, magnetic field, antigen, chemical composition, and so on. In this review, we briefly report recent progresses in light-driven materials and membranes. Photo-switching mechanisms, valved-membrane fabrication and light-driven properties are examined. Advances and perspectives of light responsive polymer membranes in biotechnology, chemistry and biology areas are discussed.

  8. Immunosensor Based on Surface Plasmon Resonance for Antigen Recognition

    Institute of Scientific and Technical Information of China (English)

    2008-01-01

    A novel immunosensor based on surface plasmon resonance(SPR)has been developed for the recognition of antigen.The sensor was designed on the basis of the fixed angle of incidence and measuring the reflected intensities in a wavelength range of 430-750 nm in real-time. An ultra-bright white light-emitting diode(LED)was used as the light source. Molecular self-assembling in solution was used to form the sensing membrane on gold substrate. It has been seen that the sensitivity of the SPR sensor with 3-mercaptopropionic acid(MPA)/protein A(SPA) sensing membrane is considerably higher than that with MPA or SPA modified Sensing membrane. The kinetic processes on the sensing membrane were studied. The human B factor(Bf), an activator of complement 3(C3), was recognized among the other antigens. This sensor can also be used for other antigen/antibody or adaptor/receptor recognition. Under optimized experimental conditions, the sensor has good selectivity, repeatability, and reversibility.

  9. Carcino-Embryonic Antigen

    International Nuclear Information System (INIS)

    Tumour marker analysis has increased our understanding of the presence of tumours in the body. Carcino-embryonic antigen, CEA, is one of the best studied tumour markers and has proved an ideal diagnostic adjuvant. It has helped in quantifying the amount of disease present in a patient and thence to make accurate prognosis on the various diagnosed ailments. At UCH, it is observed that there is an increase in cancer related ailments and therefore the need for early diagnosis is more compelling in our environment to mitigate future cost of managing advanced manifestation

  10. Cancer testis antigen and immunotherapy

    Directory of Open Access Journals (Sweden)

    Krishnadas DK

    2013-04-01

    Full Text Available Deepa Kolaseri Krishnadas, Fanqi Bai, Kenneth G Lucas Department of Pediatrics, Division of Hematology/Oncology, University of Louisville, KY, USA Abstract: The identification of cancer testis (CT antigens has been an important advance in determining potential targets for cancer immunotherapy. Multiple previous studies have shown that CT antigen vaccines, using both peptides and dendritic cell vaccines, can elicit clinical and immunologic responses in several different tumors. This review details the expression of melanoma antigen family A, 1 (MAGE-A1, melanoma antigen family A, 3 (MAGE-A3, and New York esophageal squamous cell carcinoma-1 (NY-ESO-1 in various malignancies, and presents our current understanding of CT antigen based immunotherapy. Keywords: cancer testis antigens, immunotherapy, vaccine

  11. Plasma membranes from insect midgut cells

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    Walter R. Terra

    2006-06-01

    Full Text Available Plasma membranes from insect midgut cells are separated into apical and basolateral domains. The apical domain is usually modified into microvilli with a molecular structure similar to other animals. Nevertheless, the microvillar structure should differ in some insects to permit the traffic inside them of secretory vesicles that may budd laterally or pinch-off from the tips of microvilli. Other microvillar modifications are associated with proton-pumping or with the interplay with an ensheathing lipid membrane (the perimicrovilllar membrane observed in the midgut cells of hemipterans (aphids and bugs. The perimicrovillar membranes are thought to be involved in amino acid absorption from diluted diets. The microvillar and perimicrovillar membranes have densities (and protein content that depend on the insect taxon. The role played by the microvillar and perimicrovillar proteins in insect midgut physiology is reviewed here trying to provide a coherent picture of data and highlighting further research areas.As membranas plasmáticas das células intestinais dos insetos apresentam um domínio apical e outro basal. O domínio apical é geralmente modificado em microvilosidades com organização molecular similar a de outros animais, embora possam diferir naqueles insetos que apresentam vesículas secretoras em trânsito que brotam lateralmente ou destacam-se das extremidades das microvilosidades. Outras modificações microvilares estão associadas a bombeamento de prótons ou a interrelações com uma membrana lipídica (a membrana perimicrovilar que reveste as microvilosidades de células intestinais de hemípteros (pulgões e percevejos. Admite-se que as membranas perimicrovilares estejam envolvidas na absorção de aminoácidos a partir de dietas diluídas. As membranas microvilares e perimicrovilares tem densidades distintas (e conteúdo protéico que dependem do táxon do inseto. O papel desempenhado pelas proteínas microvilares e

  12. Human leucocyte antigens in tympanosclerosis.

    Science.gov (United States)

    Dursun, G; Acar, A; Turgay, M; Calgüner, M

    1997-02-01

    This study was designed to evaluate the association between certain HLA antigens and tympanosclerosis. The serum concentrations of HLA antigens were measured by a microlymphocytotoxicity technique in patients with tympanosclerosis and compared with a healthy control group. The serum levels of HLA-B35 and -DR3 were significantly higher in the patients with tympanosclerosis. This result suggests that certain types of HLA antigens may play an important role as an indicator or mediator in the pathogenesis of tympanosclerosis. PMID:9088683

  13. Expression of apical Na(+)-L-glutamine co-transport activity, B(0)-system neutral amino acid co-transporter (B(0)AT1) and angiotensin-converting enzyme 2 along the jejunal crypt-villus axis in young pigs fed a liquid formula.

    Science.gov (United States)

    Yang, Chengbo; Yang, Xiaojian; Lackeyram, Dale; Rideout, Todd C; Wang, Zirong; Stoll, Barbara; Yin, Yulong; Burrin, Douglas G; Fan, Ming Z

    2016-06-01

    Gut apical amino acid (AA) transport activity is high at birth and during suckling, thus being essential to maintain luminal nutrient-dependent mucosal growth through providing AA as essential metabolic fuel, substrates and nutrient stimuli for cellular growth. Because system-B(0) Na(+)-neutral AA co-transporter (B(0)AT1, encoded by the SLC6A19 gene) plays a dominant role for apical uptake of large neutral AA including L-Gln, we hypothesized that high apical Na(+)-Gln co-transport activity, and B(0)AT1 (SLC6A19) in co-expression with angiotensin-converting enzyme 2 (ACE2) were expressed along the entire small intestinal crypt-villus axis in young animals via unique control mechanisms. Kinetics of Na(+)-Gln co-transport activity in the apical membrane vesicles, prepared from epithelial cells sequentially isolated along the jejunal crypt-villus axis from liquid formula-fed young pigs, were measured with the membrane potential being clamped to zero using thiocyanate. Apical maximal Na(+)-Gln co-transport activity was much higher (p < 0.05) in the upper villus cells than in the middle villus (by 29 %) and the crypt (by 30 %) cells, whereas Na(+)-Gln co-transport affinity was lower (p < 0.05) in the upper villus cells than in the middle villus and the crypt cells. The B(0)AT1 (SLC6A19) mRNA abundance was lower (p < 0.05) in the crypt (by 40-47 %) than in the villus cells. There were no significant differences in B(0)AT1 and ACE2 protein abundances on the apical membrane among the upper villus, the middle villus and the crypt cells. Our study suggests that piglet fast growth is associated with very high intestinal apical Na(+)-neutral AA uptake activities via abundantly co-expressing B(0)AT1 and ACE2 proteins in the apical membrane and by transcribing the B(0)AT1 (SLC6A19) gene in the epithelia along the entire crypt-villus axis. PMID:26984322

  14. Do Parameters Of Irradiation Influences The Apical Sealing Of Er:YAG Laser Apicetomies?

    Science.gov (United States)

    Marques, Aparecida Maria Cordeiro; dos Santos, Jean Nunes; Pinheiro, Antonio Luiz Barbosa

    2011-08-01

    Failures on the sealing of the tooth apex are responsible for many failures of apical surgeries. The Er:YAG laser has been proposed as an alternative for the use of rotator instruments on surgical endodontics. 12 human extracted canines had root endodontic treatment being the apical limit was set at 1mm before the apical foramen and were distributed into 2 groups. On group I, apicectomy was performed with the Er:YAG laser (250 mJ/15 Hz). Apical cut was performed of perpendicular mode with 3mm from the apical foramen. On Group II, the same procedures and the same sequence as above was used, varying only the parameters of the Er:YAG laser (400 mJ/6 Hz). The specimens were divided into groups and fixed, by the cervical third, on wax. Impermeabilization of the residual root apical third was performed following the same procedures used in the cervical third but the residual apex was left free from the impermeabilization. After that, the roots were immersed in a 2% methylene blue solution and placed in a bacteriological oven for 48 h. The segments were visually observed and the one showing greatest level of dye leakage was selected and kept in individual container and coded accordingly. Apical staining was measured using a stereoscopic magnifying glass; a compass; and caliper. The results showed that Group I showed significantly different higher mean level of dye leakage (5.67±4.9, p<0.05). There was a significantly difference between the groups. It is concluded that the apicectomies carried out with 400 mJ/6 Hz showed the smallest infiltration value.

  15. Prevalence of Weak D Antigen In Western Indian Population

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    Tanvi Sadaria

    2015-12-01

    Full Text Available Introduction: Discovery of Rh antigens in 1939 by Landsteiner and Weiner was the revolutionary stage in blood banking. Of these antigens, D, which decides Rh positivity or negativity, is the most antigenic. A problem is encountered when an individual has a weakened expression of D (Du, i.e., fewer numbers of D antigens on red cell membrane. Aims and Objectives: To know the prevalence of weak D in Indian population because incidence varies in different population. To determine the risk of alloimmunization among Rh D negative patients who receives the blood of weak D positive donors. Material and Methods: Rh grouping of 38,962 donors who came to The Department of Immunohematology and Blood Transfusion of Civil Hospital, Ahmedabad from 1st January 2013 to 30th September 2014 was done using the DIAGAST (Automated Grouping. The samples that tested negative for D antigen were further analysed for weak D (Du by indirect antiglobulin test using blend of Ig G and Ig M Anti D. This was done using Column agglutination method in ID card (gel card. Results: The total number of donors studied was 38,962. Out of these 3360(8.6% were tested Rh D negative. All Rh D negative donors were tested for weak D (Du. 22 (0.056% of total donors and 0.65% of Rh negative donors turned out to be weak D (Du positive. Conclusion: The prevalence of weak D (Du in Western Indian population is 0.056 %, So the risk of alloimmunization in our setting due to weak D (Du antigen is marginal. But, testing of weak D antigen is necessary in blood bank because weak D antigen is immunogenic and can produce alloimmunization if transfused to Rh D negative subjects.

  16. Lewis (y) Antigen Overexpression Increases the Expression of MMP-2 and MMP-9 and Invasion of Human Ovarian Cancer Cells

    OpenAIRE

    Shulan Zhang; Masao Iwamori; Changzhi Wang; Yifei Wang; Chuan Liu; Song Gao; Lili Gao; Bei Lin; Limei Yan

    2010-01-01

    Lewis (y) antigen is a difucosylated oligosaccharide present on the plasma membrane, and its overexpression is frequently found in human cancers and has been shown to be associated with poor prognosis. Our previous studies have shown that Lewis (y) antigen plays a positive role in the process of invasion and metastasis of ovarian cancer cells. However, the mechanisms by which Lewis (y) antigen enhances the invasion and tumor metastasis are still unknown. In this study, we established a stable...

  17. Síndrome do balonamento apical secundário ao uso abusivo de descongestionante nasal Síndrome del abombamiento apical secundario al uso abusivo de descongestionante nasal Apical ballooning syndrome secondary to nasal decongestant abuse

    Directory of Open Access Journals (Sweden)

    Ricardo Wang

    2009-11-01

    Full Text Available Descrevemos um caso típico de síndrome do balonamento apical em uma paciente octogenária com alteração eletrocardiográfica, de contratilidade do ventrículo esquerdo, e que apresentou recuperação da função ventricular. A paciente é portadora de rinite alérgica e fez uso excessivo de descongestionante nasal horas antes do episódio da dor.Describimos un caso típico de síndrome de abombamiento apical en una paciente octogenaria con alteración electrocardiográfica, de contractilidad del ventrículo izquierdo, y que presentó recuperación de la función ventricular. La paciente es portadora de rinitis alérgica y hace uso excesivo de descongestionante nasal horas antes del episodio del dolor.We describe a typical case of apical ballooning syndrome in an octogenarian female patient with left ventricular wall motion abnormality on electrocardiography, whose ventricular function returned to normal. The patient has allergic rhinitis and had used nasal decongestant excessively a few hours prior to the episode of pain.

  18. Membrane Biophysics

    CERN Document Server

    Ashrafuzzaman, Mohammad

    2013-01-01

    Physics, mathematics and chemistry all play a vital role in understanding the true nature and functioning of biological membranes, key elements of living processes. Besides simple spectroscopic observations and electrical measurements of membranes we address in this book the phenomena of coexistence and independent existence of different membrane components using various theoretical approaches. This treatment will be helpful for readers who want to understand biological processes by applying both simple observations and fundamental scientific analysis. It provides a deep understanding of the causes and effects of processes inside membranes, and will thus eventually open new doors for high-level pharmaceutical approaches towards fighting membrane- and cell-related diseases.

  19. The differences in short- and long-term varicella-zoster virus (VZV) immunoglobulin G levels following varicella vaccination of healthcare workers measured by VZV fluorescent-antibody-to-membrane-antigen assay (FAMA), VZV time-resolved fluorescence immunoassay and a VZV purified glycoprotein enzyme immunoassay.

    Science.gov (United States)

    Maple, P A C; Haedicke, J; Quinlivan, M; Steinberg, S P; Gershon, A A; Brown, K E; Breuer, J

    2016-08-01

    Healthcare workers (HCWs) reporting no history of varicella frequently receive varicella vaccination (vOka) if they test varicella-zoster virus (VZV) immunoglobulin G (IgG) negative. In this study, the utilities of VZV-IgG time-resolved fluorescence immunoassay (VZV-TRFIA) and a commercial VZV-IgG purified glycoprotein enzyme immunoassay (gpEIA) currently used in England for confirming VZV immunity have been compared to the fluorescent-antibody-to-membrane-antigen assay (FAMA). A total of 110 HCWs received two doses of vOka vaccine spaced 6 weeks apart and sera collected pre-vaccination (n = 100), at 6 weeks post-completion of vaccination (n = 86) and at 12-18 months follow-up (n = 73) were analysed. Pre-vaccination, by FAMA, 61·0% sera were VZV IgG negative, and compared to FAMA the sensitivities of VZV-TRFIA and gpEIA were 74·4% [95% confidence interval (CI) 57·9-87·0] and 46·2% (95% CI 30·1-62·8), respectively. Post-completion of vaccination the seroconversion rate by FAMA was 93·7% compared to rates of 95·8% and 70·8% determined by VZV-TRFIA and gpEIA, respectively. At 12-18 months follow-up seropositivity rates by FAMA, VZV-TRFIA and gpEIA were 78·1%, 74·0% and 47·9%, respectively. Compared to FAMA the sensitivities of VZV-TRFIA and gpEIA for measuring VZV IgG following vaccination were 96·4% (95% CI 91·7-98·8) and 74·6% (95% CI 66·5-81·6), respectively. Using both FAMA and VZV-TRFIA to identify healthy adult VZV susceptibles and measure seroconversion showed that vOka vaccination of HCWs is highly immunogenic. PMID:27018820

  20. In vitro study of apical leakage of root canals after different root apex resection techniques

    Directory of Open Access Journals (Sweden)

    André Augusto Franco MARQUES

    2008-12-01

    Full Text Available Objective:The aim of the present study was to evaluate in vitro apicalmarginal leakage of root canals after three root apex resectiontechniques.Material and methods: Thirty maxillary canines wereprepared chemo-mechanically to a size 40 master apical file, andenlarged by using Gates Glidden burs 2, 3 and 4. Teeth were obturatedby Tagger thermomechanical compaction technique and then dividedinto three groups (n = 10. The first group (GI was 3 mm apicalresected with Endo Z in high speed handpiece. The specimens of secondgroup (GII were 3 mm apical weared with Endo Z, and the third group(GIII had 3 mm apical resected with Er:YAG laser (500 impulse, 12pps and 300 mJ energy. After the apical sections, IRM cement wasplaced into the root-end cavity and teeth of each group were immersedin India ink for 5 days and submitted to decalcification and clarifying for marginal apical microleakage visualization with microscope.Results: The results showed statistically difference (p < 0.01 between GI and the other groups.Conclusion: The root apex cut using Endo Z promotes the lowest values of microleakage when used in apicoectomy.

  1. The Effect of Smear Layer on Apical Seal of Endodontically Treated Teeth

    Directory of Open Access Journals (Sweden)

    A Farhad

    2004-06-01

    Full Text Available Background: The purpose of this study was to evaluate the effect of smear layer on apical seal of endodontically treated teeth. Methods: Sixty extracted single–rooted human teeth were randomly divided into two experimental groups (n=25 and two control groups (n=5. The teeth were instrumented with K–type files to size 40 and then flared. Apical patency was ensured in all teeth. 5% sodium hypochlorite and 17% Ethylenediamine tetraacetic acid was used as irrigants to remove the smear layer in the second experimental group. The experimental groups were obturated by laterally condensed gutta – percha with Roth 801 sealer. The positive control group was obturated with gutta – percha without sealer. The root surfaces were then coated with nail polish and sticky wax except for the apex in the experimental groups and positive control group. The roots were completely covered in the negative control group. The samples were then immersed in India ink for 1 week at 37°C. Results: The statistical analysis of the results showed that the apical leakage was significantly increased in obturated canals with smear layer. Conclusion: The removal of smear layer might improve the long term apical seal and success of endodontically treated teeth. Keywords: Apical Seal, EDTA, Removal, Sodium Hypochlorite, Smear Layer.

  2. Midventricular Hypertrophic Cardiomyopathy with Apical Aneurysm: Potential for Underdiagnosis and Value of Multimodality Imaging

    Directory of Open Access Journals (Sweden)

    Archana Sivanandam

    2016-01-01

    Full Text Available We illustrate a case of midventricle obstructive HCM and apical aneurysm diagnosed with appropriate use of multimodality imaging. A 75-year-old African American woman presented with a 3-day history of chest pain and dyspnea with elevated troponins. Her electrocardiogram showed sinus rhythm, left atrial enlargement, left ventricular hypertrophy, prolonged QT, and occasional ectopy. After medical therapy optimization, she underwent coronary angiography for an initial diagnosis of non-ST segment elevation myocardial infarction. Her coronaries were unremarkable for significant disease but her left ventriculogram showed hyperdynamic contractility of the midportion of the ventricle along with a large dyskinetic aneurysmal apical sac. A subsequent transthoracic echocardiogram provided poor visualization of the apical region of the ventricle but contrast enhancement identified an aneurysmal pouch distal to the midventricular obstruction. To further clarify the diagnosis, cardiac magnetic resonance imaging with contrast was performed confirming the diagnosis of midventricular hypertrophic cardiomyopathy with apical aneurysm and fibrosis consistent with apical scar on delayed enhancement. The patient was medically treated and subsequently underwent elective implantable defibrillator placement in the ensuing months for recurrent nonsustained ventricular tachycardia and was initiated on prophylactic oral anticoagulation with warfarin for thromboembolic risk reduction.

  3. Reguladores vegetais na quebra da dominância apical de mamoeiro (Carica papaya L. Plant growth regulators on breaking apical dominance in papaya plants (Carica papaya L.

    Directory of Open Access Journals (Sweden)

    Elizabeth Orika Ono

    2004-08-01

    Full Text Available O trabalho avaliou os efeitos de reguladores vegetais sobre a quebra da dominância apical de mamoeiro (Carica papaya L. cv. Improved Sunrise Solo. A aplicação dos reguladores vegetais foi iniciada quando as plantas tinham seis meses de idade, totalizando três aplicações, a intervalos de sete dias, constando dos seguintes tratamentos: T1- água (testemunha; T2- GA3 250 mg L-1; T3- GA3 500 mg L-1; T4- benziladenina (BA 250 mg L-1; T5- BA 500 mg L-1; T6- GA3 125 mg L-1 + BA 125 mg L-1; T7- GA3 250 mg L-1 + BA 250 mg L-1. Esses tratamentos foram acompanhados da remoção ou não da gema apical. Os resultados mostraram que plantas tratadas com GA3 + BA a 125 e 250 mg L-1, com e sem a remoção da gema apical, apresentaram maior número de brotações que a testemunha, a qual não apresentou nenhuma brotação das gemas laterais.The objective of this work was to study the effects of gibberelin and cytokinin on breaking apical dominance and axillary buds growth of the Carica papaya L. Papaya plants sprayed three times within a period of 7 days, with the following treatments: T1- water (control, T2- GA3 250 mg L-1, T3- GA3 500 mg L-1, T4- benzyladenine (BA 250 mg L-1, T5- BA 500 mg L-1, T6- GA3 125 mg L-1 + BA 125 mg L-1, T7- GA3 250 mg L-1 + BA 250 mg L-1. The treatments were carried out with or without removal of the apical buds. After 36 days of the last plant growth regulators application was observed lateral bud numbers, lateral buds length and lateral buds diameter. The results showed that there were a higher number of shoots in plants treated with GA3 at 125 mg L-1 + BA at 125 mg L-1 and GA3 at 250mg L-1 + BA at 250 mg L-1. The removal of the apical bud helped the sprouting of the papaya plants.

  4. Elevated cAMP increases aquaporin-3 plasma membrane diffusion

    DEFF Research Database (Denmark)

    Marlar, Saw; Christensen, Eva Arnspang; Koffman, Jennifer Skaarup;

    2014-01-01

    .05)]. Immunoelectron microscopy showed no obvious difference in AQP3-EGFP expression levels or localization in the plasma membrane upon forskolin stimulation. Thus AQP3-EGFP diffusion is altered upon increased cAMP, which may correspond to basolateral adaptations in response to the increased apical water readsorption......Regulated urine concentration takes place in the renal collecting duct upon arginine vasopressin (AVP) stimulation, where subapical vesicles containing aquaporin-2 (AQP2) are inserted into the apical membrane instantly increasing water reabsorption and urine concentration. The reabsorped water...... be short-term regulated via changes in protein-protein interactions, incorporation into lipid rafts, and/or changes in steady-state turnover, which could result in changes in the diffusion behavior of AQP3. Thus we measured AQP3 diffusion coefficients upon stimulation with the AVP mimic forskolin to reveal...

  5. Estudio sobre la adaptación apical de conos maestros de gutapercha en conductos radiculares instrumentados con limas Lightspeed

    OpenAIRE

    Pumarola Suñé, José; García Campaña, Ana M.; Brau Aguadé, Esteban; Canalda Sahli, Carlos

    2000-01-01

    El objetivo de este trabajo fue establecer la coincidencia entre el calibre del cono maestro apical y la lima maestra apical, así como medir cualitativamente la adaptación de diferentes marcas de conos de gutapercha en el tercio apical de conductos radiculares instrumentados con limas Lightspeed. Se instrumentaron 100 conductos de molares naturales extraídos con el sistema Lightspeed. Las muestras se distribuyeron aleatoriamente en cinco grupos. Cada grupo fue obturado mediante condensación l...

  6. Tissue reaction to Endométhasone sealer in root canal fillings short of or beyond the apical foramen

    Directory of Open Access Journals (Sweden)

    Patrícia Suzuki

    2011-10-01

    Full Text Available OBJECTIVE: This study evaluated the response of periapical tissues to the endodontic sealer Endométhasone in root canal fillings short of or beyond the apical foramen. MATERIAL AND METHODS: Twenty root canals of premolars and incisors of 2 mongrel dogs were used. After coronal access and pulp extirpation, the canals were instrumented up to a size 55 K-file and the apical cemental barrier was penetrated with a size 15 K-file to obtain a main apical foramen, which was widened to a size 25 K-file. The canals were irrigated with saline at each change of file. The root canals were obturated either short of or beyond the apical foramen by the lateral condensation of gutta-percha and Endométhasone, originating 2 experimental groups: G1: Endométhasone/short of the apical foramen; G2: Endométhasone/beyond the apical foramen. The animals were killed by anesthetic overdose 90 days after endodontic treatment. The individual roots were obtained and serial histological sections were prepared for histomorphological analysis (H&E and Brown & Brenn techniques under light microscopy. The following parameters were examined: closure of the apical foramen of the main root canal and apical opening of accessory canals, apical cementum resorption, intensity of the inflammatory infiltrate, presence of giant cells and thickness and organization of the apical periodontal ligament. Each parameter was scored 1 to 4, 1 being the best result and 4 the worst. Data were analyzed statistically by the Wilcoxon nonparametric tests (p=0.05. RESULTS: Comparing the 2 groups, the best result (p<0.05 was obtained with root canal filling with Endométhasone short of the apical foramen but a chronic inflammatory infiltrate was present in all specimens. CONCLUSIONS: Limiting the filling material to the root canal space apically is important to determine the best treatment outcome when Endométhasone is used as sealer.

  7. Antigen antibody interactions

    CERN Document Server

    DeLisi, Charles

    1976-01-01

    1. 1 Organization of the Immune System One of the most important survival mechanisms of vertebrates is their ability to recognize and respond to the onslaught of pathogenic microbes to which they are conti- ously exposed. The collection of host cells and molecules involved in this recognition­ 12 response function constitutes its immune system. In man, it comprises about 10 cells 20 (lymphocytes) and 10 molecules (immunoglobulins). Its ontogenic development is c- strained by the requirement that it be capable of responding to an almost limitless variety of molecular configurations on foreign substances, while simultaneously remaining inert to those on self components. It has thus evolved to discriminate, with exquisite precision, between molecular patterns. The foreign substances which induce a response, called antigens, are typically large molecules such as proteins and polysaccharides. The portions of these with which immunoglobulins interact are called epitopes or determinants. A typical protein epitope m...

  8. Gibberellin Is Involved in the Regulation of Cell Death-mediated Apical Senescence in G2 Pea

    Institute of Scientific and Technical Information of China (English)

    2007-01-01

    Senescence is the process of programmed degradation.The G2 line of pea exhibits apical senescence-delaying phenotype under short-day(SD)conditions,but the mechanism regulating the apical senescence is still largely unknown.Gibberellin (GA)was proved to be able to delay this apical senescence phenotype in G2 pea grown under long-day(LD) conditions.Here we show that the initiation of cell death signals in the tarminal floral meristem was involved in the regulation of apical senescence in pea plants.SD signals prevented the formation of the cell death region in the apical mersitem.Moreovsr.GA3 treatment could effectively inhibit the occurrence of cell death-mediated apical senescence in LD-grown apical buds.Therefore,our data suggest that the prevention of apical senescence in SD-grown G2 pea through GA3 treatment may be largely responsible for the regulation of occurrence of the DNA fragmentation in apical meristem.

  9. Membrane potential plays a dual role for chloride transport across toad skin

    DEFF Research Database (Denmark)

    Larsen, Erik Hviid; Rasmussen, B E

    1983-01-01

    -dependent currents are not caused by a trivial Goldmand-type rectification and ion redistributions following transepithelial potential pertubations. Extended with a dynamic Cl- permeability in the apical membrane according to a Hodgkin-Huxley kinetic scheme, the model predicts voltage clamp data which closely...

  10. Polarized membrane traffic and cell polarity development is dependent on dihydroceramide synthase-regulated sphinganine turnover

    NARCIS (Netherlands)

    van Ijzendoorn, SCD; van der Wouden, JM; Liebisch, G; Schmitz, G; Hoekstra, D

    2004-01-01

    Sphingoid bases have been implicated in various cellular processes including cell growth, apoptosis and cell differentiation. Here, we show that the regulated turnover of sphingoid bases is crucial for cell polarity development, i.e., the biogenesis of apical plasma membrane domains, in well-differe

  11. The apical oblique view of the clavicle: Its usefulness in neonatal and childhood trauma

    International Nuclear Information System (INIS)

    We analyzed clavicular radiographs of 26 patients with a history of trauma. The apical oblique projection of the clavicle was obtained with the injured side of the patient angled 450 towards the X-ray tube and a 200 cephalad angulation of the X-ray beam. This view proved to be more informative than the routine apical anteroposterior projection. It is especially effective in detecting nondisplaced fractures of the middle third of the clavicle in neonates and children. To verify our findings, we obtained apical anteroposterior and oblique radiographs of a specimen adult of the X-ray beam, the measurements of the projected lengths of the anatomical specimen, especially those of the middle portion of the clavicle, were very close to the corresponding anatomical lengths. (orig.)

  12. Redistribution and modulation of Gross murine leukemia virus antigens induced by specific antibodies.

    Science.gov (United States)

    Ioachim, H L; Sabbath, M

    1979-01-01

    Gross murine leukemia virus (G-MuLV)-induced rat leukemia cells in tissue culture replicate G-MuLV, express strong virus-associated membrane antigenicity, and are consistently killed by specific antibodies and complement in cytotoxicity tests. To explore the effect of specific antibodies, rat anti-G-MuLV antisera were added to the cultures of leukemia cells for variable periods of time. Redistribution of virus particles as well as of membrane virus antigens in the form of polar patches and caps was observed by electron microscopy, indirect immunofluorescence, and immunoelectron microscopy. Substantial decreases in cytotoxicity indexes accompanied these changes. The antigen modulation induced by anti-G-MuLV antibodies in vitro paralleled similar changes obtained in vivo by transplanttion of leukemia cells in rats with high anti-G-MuLV antibody titers. The importance of antigen modulation in this system resides in its direct relationship with the malignant potential of the leukemia cells.

  13. Interactions between HIV-1 Neutralizing Antibodies and Model Lipid Membranes imaged with AFM

    Science.gov (United States)

    Zauscher, Stefan; Hardy, Gregory; Alam, Munir; Shapter, Joseph

    2012-02-01

    Lipid membrane interactions with rare, broadly neutralizing antibodies (NAbs), 2F5 and 4E10, play a critical role in HIV-1 neutralization. Our research is motivated by recent immunization studies that have shown that induction of antibodies that avidly bind the gp41-MPER antigen is not sufficient for neutralization. Rather, it is required that antigen designs induce polyreactive antibodies that recognize MPER antigens as well as the viral lipid membrane. However, the mechanistic details of how membrane properties influence NAb-lipid and NAb-antigen interactions remain unknown. Furthermore, it is well established that the native viral membrane is heterogeneous, representing a mosaic of lipid rafts and protein clustering. However, the size, physical properties, and dynamics of these regions are poorly characterized and their potential roles in HIV-1 neutralization are also unknown. To understand how membrane properties contribute to 2F5/4E10 membrane interactions, we have engineered biomimetic supported lipid bilayers (SLBs) and use atomic force microscopy to visualize membrane domains, antigen clustering, and antibody-membrane interactions at sub-nanometer z-resolution. Our results show that localized binding of HIV-1 antigens and NAbs occur preferentially with the most fluid membrane domain. This supports the theory that NAbs may interact with regions of low lateral lipid forces that allow antibody insertion into the bilayer.

  14. Membrane Processes.

    Science.gov (United States)

    Pellegrin, Marie-Laure; Sadler, Mary E; Greiner, Anthony D; Aguinaldo, Jorge; Min, Kyungnan; Zhang, Kai; Arabi, Sara; Burbano, Marie S; Kent, Fraser; Shoaf, Robert

    2015-10-01

    This review, for literature published in 2014, contains information related to membrane processes for municipal and industrial applications. This review is a subsection of the Treatment Systems section of the annual Water Environment Federation literature review and covers the following topics: pretreatment, membrane bioreactor (MBR) configuration, design, nutrient removal, operation, industrial treatment, fixed film and anaerobic membrane systems, reuse, microconstituents removal, membrane technology advances, membrane fouling, and modeling. Other sub-sections of the Treatment Systems section that might relate to this literature review include: Biological Fixed-Film Systems, Activated Sludge and Other Aerobic Suspended Culture Processes, Anaerobic Processes, Water Reclamation and Reuse. The following sections might also have related information on membrane processes: Industrial Wastes, Hazardous Wastes, and Fate and Effects of Pollutants. PMID:26420079

  15. Membrane Processes.

    Science.gov (United States)

    Pellegrin, Marie-Laure; Burbano, Marie S; Sadler, Mary E; Diamond, Jason; Baker, Simon; Greiner, Anthony D; Arabi, Sara; Wong, Joseph; Doody, Alexandra; Padhye, Lokesh P; Sears, Keith; Kistenmacher, Peter; Kent, Fraser; Tootchi, Leila; Aguinaldo, Jorge; Saddredini, Sara; Schilling, Bill; Min, Kyungnan; McCandless, Robert; Danker, Bryce; Gamage, Neranga P; Wang, Sunny; Aerts, Peter

    2016-10-01

    This review, for literature published in 2015, contains information related to membrane processes for municipal and industrial applications. This review is a subsection of the Treatment Systems section of the annual Water Environment Federation literature review and covers the following topics: pretreatment, membrane bioreactor (MBR) configuration, design, nutrient removal, operation, industrial treatment, anaerobic membrane systems, reuse, microconstituents removal, membrane technology advances, membrane fouling, and modeling. Other sub-sections of the Treatment Systems section that might relate to this literature review include: Biological Fixed-Film Systems, Activated Sludge and Other Aerobic Suspended Culture Processes, Anaerobic Processes, Water Reclamation and Reuse. The following sections might also have related information on membrane processes: Industrial Wastes, Hazardous Wastes, and Fate and Effects of Pollutants. PMID:27620084

  16. Estudio de la filtración apical de cuatro cementos de obturación

    OpenAIRE

    Roig Cayón, Miguel; Ribot Porta, J. de; Jané Noblom, L.; Canalda Sahli, Carlos

    1996-01-01

    Los autores estudian in vitro la microfiltración apical con cuatro cementos de obturación, mediante la utilización de un colorante y técnicas de diafanización. Analizan dos cementos a base de hidróxido de calcio, y uno de ionómero de vidrio, comparándolos con un cemento clásico a base de óxido de zinc-eugenol. La filtración apical del cemento sellador Apexit fue significativamente superior a la de los cementos Sealapex y Endomethasone. No se observaron diferencias significativas entre los dem...

  17. Apical Ballooning Syndrome (Takotsubo Cardiomyopathy after Permanent Dual-Chamber Pacemaker Implantation

    Directory of Open Access Journals (Sweden)

    Armando Gardini

    2012-01-01

    Full Text Available Apical ballooning syndrome, also called takotsubo cardiomyopathy, has been recently reported. It may mimic acute myocardial infarction and is typically observed in postmenopausal women after stressful events. A 75-year-old female after permanent dual chamber pacemaker implant complained of chest pain with repolarization alterations suggesting acute myocardial ischemia. Echocardiography showed a left ventricle with akinesia of the apical portions and reduced global systolic function. The patient was treated with antithrombotic agents and intravenous nitrates. No coronary lesions were found at angiography. At ventriculography, a typical takotsubo-like shape of the left ventricle was observed. The clinical and echocardiographic picture normalized at discharge.

  18. Reasons for Apical Surgery Treatment in an Underserved New York City Population.

    Science.gov (United States)

    Hasselgren, Gunnar; Patel, Pratik; Alhassany, Hashim; Kunzel, Carol

    2016-04-01

    Endodontic retreatment often involves remaking restorations. The total cost may steer the treatment towards surgery. The aim of this study was to retrospectively record the reasons for performing apical surgery in an economically deprived patient population. The clinical reasons (59%) for apical surgery were most common, but the nonclinical (financial) reasons (41%) emerged as a major cause. The finding that 41% of the apicoectomies were performed because of nonclinical constraints is a high figure and may not reflect the situation generally. Still, economic factors potentially play a major role in the selection of surgical versus nonsurgical endodontic retreatment. PMID:27348949

  19. SEALING ABILITY OF GRAY MTA ANGELUS™, CPM™ AND MBPC USED AS APICAL PLUGS

    OpenAIRE

    Fernando Accorsi Orosco; Clovis Monteiro Bramante; Roberto Brandão Garcia; Norberti Bernadineli; Ivaldo Gomes de Moraes

    2008-01-01

    This study evaluated the sealing ability of apical plugs fabricated with gray MTA AngelusTM sealer, CPM TM sealer and MBPc sealer. The root canals of 98 extracted single-rooted human teeth were instrumented with #5 to #1 Gates Glidden drills according to the crown-down technique until the #1 drill could pass through the apical foramen. The specimens were then prepared with K-files, starting with an ISO 50 until an ISO 90 could be visualized 1 mm beyond the apex. After root canal preparation, ...

  20. Multicomponent membranes

    Science.gov (United States)

    Kulprathipanja, Santi; Kulkarni, Sudhir S.; Funk, Edward W.

    1988-01-01

    A multicomponent membrane which may be used for separating various components which are present in a fluid feed mixture comprises a mixture of a plasticizer such as a glycol and an organic polymer cast upon a porous organic polymer support. The membrane may be prepared by casting an emulsion or a solution of the plasticizer and polymer on the porous support, evaporating the solvent and recovering the membrane after curing.

  1. [Antigenic response against PPD and antigen 60 in tubercular patients: single antigen versus the combined test].

    Science.gov (United States)

    Máttar, S; Broquetas, J M; Gea, J; Aran, X; el-Banna, N; Sauleda, J; Torres, J M

    1992-05-01

    We analyze serum samples from 70 patients with pulmonary tuberculosis and 50 healthy individuals. The antigenic activity (IgG) against protein purified antigen (PPD) and antigen 60 (A60) from M. tuberculosis. Thirteen patients were also HIV infected, and three patients had AIDS defined by the presence of disseminated tuberculosis. The test using antigen alone showed a 77% sensitivity and 74% specificity when PPD is used. When A60 was used, both values improved (81% sensitivity, 94% specificity). The use of a combined test (PPD and A60) improves the sensitivity (89%) but reduces the specificity (82%). The HIV infected patients showed similar responses to those of other patients. The combined use of different antigens might be useful for diagnosing tuberculosis. PMID:1390996

  2. [Mucose associated lymphoid tissue. Antigen presenting cells].

    Science.gov (United States)

    Luzardo-Baptista, Mario J; Luzardo, José Rafael

    2013-12-01

    We studied samples of normal and abnormal human mucosae, including oral tissue and uterine cervix, using electron microscopy. Special attention was given to the functions and mechanisms of defense carried out by the epithelial (EC) and dendritic cells (DC). Activated epithelial cells posses the capacity to uptake and process antigens, in order to present them, subsequently, to the dendritic cells. The structures and elements of the cells intervening on this function are: micropinocytic vesicles, multivesicular bodies, lysosomes, phagosomes, clathrin-covered vesicles, dense granules covered by a unit membrane, granules with onion likes leaves, microbodies, and dense granules with acid phosphatase activity. When they first arrive within the epithelial layers, the DC are clear with long cytoplasmic projections, which later become short, and the density of their cytoplasm increases. They possess mycropinocytic vesicles, some clathrine-covered vesicles, lysososmes and Birbeck granules. At this moment, they are known as Langerhans cells. EC and DC present many surface folds rich in micropynocytic vesicles. Between EC and DC there are many contacts (close junctions or tight junctions), through which antigens, phagocitized and processed by the EC, are given to the DC. These cells join them to major histocompatibility complex molecules or to other molecules with similar functions (CD1). Then the Langerhans cells travel to the lymphatic node to activate T cells and continue the immunologic task. So, in this way, both the EC and the DC are a link between the natural and the acquired immunological mechanisms. PMID:24502183

  3. COLONOSCOPY AND CARCINOEMBRYONIC ANTIGEN VARIATIONS

    Directory of Open Access Journals (Sweden)

    Rita G SOUSA

    2014-03-01

    Full Text Available Context Colonoscopy is essential for synchronous and metachronous cancer detection. Carcinoembryonic antigen is a colorectal cancer tumor marker, important as a follow-up tool in patients with previous colorectal cancer. False-positive carcinoembryonic antigen elevation results in multiples exams and in patient anxiety. In literature, there is reference to transient carcinoembryonic antigen increase with colonoscopy. Objective To evaluate the influence of bowel preparation and colonoscopy in carcinoembryonic antigen blood levels. Methods We prospectively studied subjects that underwent routine colonoscopy in our institution. Blood samples were collected (1 before bowel cleaning, (2 before colonoscopy and (3 immediately after colonoscopy. Blood carcinoembryonic antigen levels were determined by “Sandwich” immunoassay. The statistical methods used were the paired t-test and ANOVA. Results Thirty-seven patients (22M/15F were included; age range 28-84 (mean 56 years. Mean carcinoembryonic antigen values were 1.9, 2 and 1.8 for (1, (2 and (3, respectively. An increase in value (2 compared with (1 was observed in 20/37 patients (P = 0.018, mainly in younger patients and in patients requiring more endoluminal interventions. In 29/37 patients, the CEA value decreased from (2 to (3 (P = 1.3x10-7. Conclusions A trend for carcinoembryonic antigen increase after bowel cleaning was observed, especially in younger patients and in patients with more endoluminal interventions, but without clinical meaning.

  4. Regulation of protein synthesis and autophagy in activated dendritic cells: implications for antigen processing and presentation.

    Science.gov (United States)

    Argüello, Rafael J; Reverendo, Marisa; Gatti, Evelina; Pierre, Philippe

    2016-07-01

    Antigenic peptides presented in the context of major histocompatibility complex (MHC) molecules originate from the degradation of both self and non-self proteins. T cells can therefore recognize at the surface of surveyed cells, the self-peptidome produced by the cell itself (mostly inducing tolerance) or immunogenic peptides derived from exogenous origins. The initiation of adaptive immune responses by dendritic cells (DCs), through the antigenic priming of naïve T cells, is associated to microbial pattern recognition receptors engagement. Activation of DCs by microbial product or inflammatory cytokines initiates multiple processes that maximize DC capacity to present exogenous antigens and stimulate T cells by affecting major metabolic and membrane traffic pathways. These include the modulation of protein synthesis, the regulation of MHC and co-stimulatory molecules transport, as well as the regulation of autophagy, that, all together promote exogenous antigen presentation while limiting the display of self-antigens by MHC molecules.

  5. The endoplasmic reticulum and casein-containing vesicles contribute to milk fat globule membrane.

    Science.gov (United States)

    Honvo-Houéto, Edith; Henry, Céline; Chat, Sophie; Layani, Sarah; Truchet, Sandrine

    2016-10-01

    During lactation, mammary epithelial cells secrete huge amounts of milk from their apical side. The current view is that caseins are secreted by exocytosis, whereas milk fat globules are released by budding, enwrapped by the plasma membrane. Owing to the number and large size of milk fat globules, the membrane surface needed for their release might exceed that of the apical plasma membrane. A large-scale proteomics analysis of both cytoplasmic lipid droplets and secreted milk fat globule membranes was used to decipher the cellular origins of the milk fat globule membrane. Surprisingly, differential analysis of protein profiles of these two organelles strongly suggest that, in addition to the plasma membrane, the endoplasmic reticulum and the secretory vesicles contribute to the milk fat globule membrane. Analysis of membrane-associated and raft microdomain proteins reinforces this possibility and also points to a role for lipid rafts in milk product secretion. Our results provide evidence for a significant contribution of the endoplasmic reticulum to the milk fat globule membrane and a role for SNAREs in membrane dynamics during milk secretion. These novel aspects point to a more complex model for milk secretion than currently envisioned.

  6. Prokaryotic expression of surface membrane antigen SAG1 gene from Toxoplasma Gondii and the diagnostic value of the recombinant protein%弓形虫膜表面抗原SAG1基因的原核表达及重组蛋白的免疫诊断价值

    Institute of Scientific and Technical Information of China (English)

    王朝兰; 汤冬生; 姚湧; 汪学龙; 王业梅

    2011-01-01

    目的 探讨重组弓形虫膜表面抗原SAG1基因的表达产物-原核表达蛋白(rSAG1)用于弓形虫病的免疫诊断价值.方法 用异丙基-B-D-硫代吡喃半乳糖苷(IPTG)诱导大肠埃希菌重组质粒pET28a-SAG1(pET28a-SAG1/BL21)表达,纯化重组pET28a-SAG1/BL21弓形虫膜表面抗原SAG1基因表达产物;用弓形虫缓殖子感染的鼠血清、正常鼠血清和10例弓形虫患者血清为一抗,基因表达产物rSAG1用免疫印迹法进行鉴定,比较rSAG1在弓形虫病免疫诊断中的价值.结果 纯化重组弓形虫膜表面抗原SAG1基因后获得了相对分子质量约38.5×103的表达产物rSAG1;表面抗原SAG1可被弓形虫缓殖子感染的鼠血清所识别;10例弓形虫患者血清在免疫印迹诊断中,有4例出现了弓形虫膜表面抗原SAG1基因表达产物rSAG1.结论 rSAG1具备一定的弓形虫病的免疫诊断价值.%Objective To investigate the diagnostic value of the recombinant surface antigen 1 (rSAG1) in immunodiagnosis of toxoplasmosis. Methods Isopropyl β-D- 1 -thio-galaetopyranoside (IPTG) was used to induce the expression of recombinant plasmid pET28a-SAG1 of Escherich coli(pET28a-SAG1/BL21 ). The expression products (rSAG1) of pET28a-SAG1/BL21 were identified by Western blotting. The serum of mice infected with Toxoplasma gondii tachyzoites, normal mouse serum and the serum from 10 toxoplasma gondii patients were used as primary anti-Toxoplasma gondii antibodies, and the rSAG1 gene products were identified by Western blotting, by which the diagnostic value of rSAG1 in Toxoplasmosis was compared. Results After induction and purification, rSAG1 protein was obtained and its relative molecular mass was 38.5 × 103. The fusion protein could be recognized by the serum of mouse infected with Toxoplasma gondii tachyzoites, rSAG1 of expression products of surface membrane antigen SAG1 gene from Toxoplasma Gondii could be detected in 4 cases from 10 patients by Westem blotting.Conclusion The r

  7. Expression and characterization of a parasite-specific antigen on macrophages after infection with Leishmania donovani.

    Science.gov (United States)

    Basu, N; Kole, L; Ghosh, A; Das, P K

    1994-03-16

    A rabbit polyclonal antibody to crude soluble antigen of Leishmania donovani promastigotes recognized a determinant expressed on the surface membrane of mouse peritoneal macrophages and human monocyte derived macrophages infected in vitro. The determinant was recognized on infected macrophage surface only when F(ab')2 fragments of anti-leishmanial antiserum was employed in immunofluorescence. F(ab')2 fragments of human patient sera also could recognize the determinant. The expression of this antigen was not stage-specific for the parasite. Immunochemical analyses revealed this antigen to be of 51 kDa protein. Specific leaching of membrane proteins by trypsin showed three bands of expressed antigens of 26, 11 and 10 kDa, which in all likelihood might be arising from the 51 kDa antigen. The antigen was not expressed until 12 h of post infection, reached a maximum level at 24 h and thereafter attained a steady state level as studied upto 96 h of post infection. This type of antigen might have a great potential in immunodiagnostics and site-specific drug targeting. PMID:8078503

  8. Vaccine delivery by penetratin: mechanism of antigen presentation by dendritic cells.

    Science.gov (United States)

    Pouniotis, Dodie; Tang, Choon-Kit; Apostolopoulos, Vasso; Pietersz, Geoffrey

    2016-08-01

    Cell-penetrating peptides (CPP) or membrane-translocating peptides such as penetratin from Antennapedia homeodomain or TAT from human immunodeficiency virus are useful vectors for the delivery of protein antigens or their cytotoxic (Tc) or helper (Th) T cell epitopes to antigen-presenting cells. Mice immunized with CPP containing immunogens elicit antigen-specific Tc and/or Th responses and could be protected from tumor challenges. In the present paper, we investigate the mechanism of class I and class II antigen presentation of ovalbumin covalently linked to penetratin (AntpOVA) by bone marrow-derived dendritic cells with the use of biochemical inhibitors of various pathways of antigen processing and presentation. Results from our study suggested that uptake of AntpOVA is via a combination of energy-independent (membrane fusion) and energy-dependent pathways (endocytosis). Once internalized by either mechanism, multiple tap-dependent or independent antigen presentation pathways are accessed while not completely dependent on proteasomal processing but involving proteolytic trimming in the ER and Golgi compartments. Our study provides an understanding on the mechanism of antigen presentation mediated by CPP and leads to greater insights into future development of vaccine formulations. PMID:27138940

  9. Formation and removal of apical vapor lock during syringe irrigation: a combined experimental and Computational Fluid Dynamics approach

    NARCIS (Netherlands)

    Boutsioukis, C.; Kastrinakis, E.; Lambrianidis, T.; Verhaagen, B.; Versluis, M.; Sluis, van der L.W.M.

    2014-01-01

    Aim (i) To evaluate the effect of needle type and insertion depth, root canal size and irrigant flow rate on the entrapment of air bubbles in the apical part of a root canal (apical vapor lock) during syringe irrigation using experiments and a Computational Fluid Dynamics (CFD) model, (ii) to invest

  10. Formation and removal of apical vapor lock during syringe irrigation : a combined experimental and Computational Fluid Dynamics approach

    NARCIS (Netherlands)

    Boutsioukis, C.; Kastrinakis, E.; Lambrianidis, T.; Verhaagen, B.; Versluis, M.; van der Sluis, L. W. M.

    2014-01-01

    Aim(i) To evaluate the effect of needle type and insertion depth, root canal size and irrigant flow rate on the entrapment of air bubbles in the apical part of a root canal (apical vapor lock) during syringe irrigation using experiments and a Computational Fluid Dynamics (CFD) model, (ii) to investi

  11. Apical root resorption 6 months after initiation of fixed orthodontic appliance therapy.

    NARCIS (Netherlands)

    Smale, I.M.; Artun, J.; Behbehani, F.; Doppel, D.; Hof, M.A. van 't; Kuijpers-Jagtman, A.M.

    2005-01-01

    INTRODUCTION: Individual predisposition might be a major reason for the observed variation in apical orthodontic root resorption. If so, resorption might be expressed during the initial stages of orthodontic therapy in patients at risk. METHODS: To explore this hypothesis, we evaluated standardized,

  12. The Effect of Calcium Hydroxide As an Intracanal Medication of The Apical Microleakage

    Directory of Open Access Journals (Sweden)

    Heshmat Mohajer AR

    2011-12-01

    Full Text Available Background and Aims: Nowadays, calcium hydroxide is used as a dressing for canal sterilization and repair progression of apical lesions. The aim of this study was to investigate the effect of calcium hydroxide as an intracanal medicament on the apical microleakage of root filling.Materials and Methods: In this experimental study, 46 extracted single-rooted human teeth were instrumented with step-back technique to master apical file (MAF size 35. Specimens were randomly divided into 2 groups (n = 20. In group 1, the specimens were treated with calcium hydroxide intracanal medication; and in group 2, the samples did not receive any medication. The teeth were incubated in 100% humidity at 37°C for one week. After that, calcium hydroxide was removed using irrigation with normal saline and reaming with MAF. The root canals were obturated with gutta- percha and AH26 sealer using lateral compaction technique. Specimens were incubated in 100% humility at 37°C for 72 hours and then immersed in India ink for 1 week. Finally, the teeth were cleared and the maximum linear dye penetration was measured under a stereomicroscope at 4X magnification. The data were analyzed by T-test and Chi-square.Results: There was no significant difference between the two experimental groups (P=0.068. Conclusion: The findings of this study indicated that using calcium hydroxide as an intracanal medicament did not influence the apical microleakage after final obturation of the root canal system.

  13. Cone beam computed tomography study of apical root resorption induced by Herbst appliance

    Directory of Open Access Journals (Sweden)

    João Paulo SCHWARTZ

    2015-10-01

    Full Text Available Objective This study evaluated the frequency of root resorption during the orthodontic treatment with Herbst appliance by Cone Beam Computed Tomography (CBCT.Material and Methods The sample comprised 23 patients (11 men, 12 women; mean ages 15.76±1.75 years with Class II division 1 malocclusion, treated with Herbst appliance. CBCT was obtained before treatment (T0 and after Herbst treatment (T1. All the dental roots, except third molars, were evaluated, and apical root resorption was determined using the axial guided navigation method. Paired t-tests and Wilcoxon T Test were used to compare the dependent samples in parametric and nonparametric cases, respectively. Chi-Square Test with Yates’ correction was used to evaluate the relationship between apical root resorption and gender. Results were considered at a significance level of 5%.Results Apical resorption was detected by CBCT in 57.96% of 980 roots that underwent Herbst appliance treatment. All patients had minimal resorption and there was no statistical significance between the genders.Conclusion CBCT three-dimensional evaluation showed association between Herbst appliance and minimal apical root resorption, mostly in the anchoring teeth, without clinical significance.

  14. Pulp Revascularization in Immature Permanent Tooth with Apical Periodontitis Using Mineral Trioxide Aggregate

    Directory of Open Access Journals (Sweden)

    Katsura Saeki

    2014-01-01

    Full Text Available Mineral trioxide aggregate (MTA is a material that has been used worldwide in several clinical applications, such as apical barriers in teeth with immature apices, repair of root perforations, root-end filling, pulp capping, and pulpotomy. The purpose of this case report was to describe successful revascularization treatment of an immature mandibular right second premolar with apical periodontitis in a 9-year-old female patient. After preparing an access cavity without anesthesia, the tooth was isolated using a rubber dam and accessed. The canal was gently debrided using 5% sodium hypochlorite (NaOCl and 3% hydrogen peroxide irrigant. And then MTA was packed into the canal. X-ray photographic examination showed the dentin bridge 5 months after the revascularization procedure. Thickening of the canal wall and complete apical closure were confirmed 10 months after the treatment. In this case, MTA showed clinical and radiographic success at revascularization treatment in immature permanent tooth. The successful outcome of this case suggests that MTA is reliable and effective for endodontic treatment in the pediatric dentistry.

  15. Regenerative Endodontic Treatment versus Apical Plug in Immature Teeth: Three-Year Follow-Up.

    Science.gov (United States)

    Asgary, Saeed; Fazlyab, Mahta; Nosrat, Ali

    2016-01-01

    This report compares and evaluates the treatment outcomes of regenerative endodontic treatment and apical plug as two accepted treatment protocols in a pair of necrotic immature maxillary central incisors of a 12-year old female. The patient was referred complaining of a dull pain and swelling in her upper lip area. She had a history of trauma to the anterior maxilla two years earlier. Both teeth were clinically diagnosed with pulp necrosis and periapical radiographs revealed that separate periapical radiolucent lesions surrounded the immature apices of both teeth. The left and right incisors were treated with apical plug and regenerative endodontic treatment, respectively, using calcium-enriched mixture (CEM) cement. The patient was followed-up for three years. During this period, both teeth were clinically asymptomatic and showed complete radiographic healing of the periapical lesions. The right central incisor showed root development. No tooth discoloration was evident. Apexification by apical plug placement and pulp regeneration are both reliable treatments for immature non-vital teeth. In order to choose the right treatment the advantages of either technique should be weighed against its drawbacks. CEM cement can be successfully applied for both purposes. This biomaterial causes less discoloration of the tooth. PMID:27617375

  16. Micro-CT analyses of apical enlargement and molar root canal complexity

    DEFF Research Database (Denmark)

    Markvart, Merete; Darvann, Tron Andre; Larsen, Per;

    2012-01-01

    AIM: To compare the effectiveness of two rotary hybrid instrumentation techniques with focus on apical enlargement in molar teeth and to quantify and visualize spatial details of instrumentation efficacy in root canals of different complexity. METHODOLOGY: Maxillary and mandibular molar teeth were...

  17. Responses of Metabolites in Soybean Shoot Apices to Changing Atmospheric Carbon Dioxide Concentrations

    Directory of Open Access Journals (Sweden)

    Richard Sicher

    2012-01-01

    Full Text Available Soybean seedlings were grown in controlled environment chambers with CO2 partial pressures of 38 (ambient and 72 (elevated Pa. Five or six shoot apices were harvested from individual 21- to 24-day-old plants. Metabolites were analyzed by gas chromatography and, out of 21 compounds, only sucrose and fructose increased in response to CO2 enrichment. One unidentified metabolite, Unk-21.03 decreased up to 80% in soybean apices in response to elevated CO2. Levels of Unk-21.03 decreased progressively when atmospheric CO2 partial pressures were increased from 26 to 100 Pa. Reciprocal transfer experiments showed that Unk-21.03, and sucrose in soybean apices were altered slowly over several days to changes in atmospheric CO2 partial pressures. The mass spectrum of Unk-21.03 indicated that this compound likely contained both an amino and carboxyl group and was structurally related to serine and aspartate. Our findings suggested that CO2 enrichment altered a small number of specific metabolites in soybean apices. This could be an important step in understanding how plant growth and development are affected by carbon dioxide enrichment.

  18. Right ventricular outflow and apical pacing comparably worsen the echocardioghraphic normal left ventricle

    NARCIS (Netherlands)

    T.J.F. ten Cate (Tim); M.G. Scheffer (Michael); G.R. Sutherland (George); J.F. Verzijlbergen (Fred); N.M. van Hemel (Norbert)

    2008-01-01

    textabstractAims: A depressed left ventricular function (LVF) is sometimes observed during right ventricular apical (RVA) pacing, but any prediction of this adverse effect cannot be done. Right ventricular outflow tract (RVOT) pacing is thought to deteriorate LVF less frequently because of a more no

  19. Hormonal networks involved in apical hook development in darkness and their response to light

    Directory of Open Access Journals (Sweden)

    Maria Agustina Mazzella

    2014-02-01

    Full Text Available In darkness, the dicot seedlings produces an apical hook as result of differential cell division and extension at opposite sides of the hypocotyl. This hook protects the apical meristem from mechanical damage during seedling emergence from the soil. In darkness, gibberellins act via the DELLA-PIF (PHYTOCHROME INTERACTING FACTORs pathway, and ethylene acts via the EIN3/EIL1 (ETHYLENE INSENSITIE 3/EIN3 like 1-HLS1 (HOOKLESS 1 pathway to control the asymmetric accumulation of auxin required for apical hook formation and maintenance. These core pathways form a network with multiple points of connection. Light perception by phytochromes and cryptochromes reduces the activity of PIFs and CONSTITUTIVE PHOTOMORPHOGENIC 1 (COP1 --both required for hook formation in darkness--, lowers the levels of gibberellins, and triggers hook opening as a component of the switch between heterotrophic and photoautotrophic development. Apical hook opening is thus a suitable model to study the convergence of endogenous and exogenous signals on the control of cell division and cell growth.

  20. A novel mitochondrial ATP8 gene mutation in a patient with apical hypertrophic cardiomyopathy and neuropathy.

    NARCIS (Netherlands)

    Jonckheere, A.I.; Hogeveen, M.; Nijtmans, L.G.J.; Brand, M.A.M. van den; Janssen, A.J.M.; Diepstra, J.H.S.; Brandt, FC van den; Heuvel, L.P.W.J. van den; Hol, F.A.; Hofste, T.G.; Kapusta, L.; Dillmann, U.; Shamdeen, M.G.; Smeitink, J.A.M.; Rodenburg, R.J.T.

    2008-01-01

    PURPOSE: To identify the biochemical and molecular genetic defect in a 16-year-old patient presenting with apical hypertrophic cardiomyopathy and neuropathy suspected for a mitochondrial disorder. METHODS: Measurement of the mitochondrial energy-generating system (MEGS) capacity in muscle and enzyme

  1. Morphology of the Physiological Apical Foramen in Maxillary and Mandibular First Molars

    Science.gov (United States)

    Abarca, J.; Zaror, C.; Monardes, H.; Hermosilla, V.; Muñoz, C.; Cantin, M.

    2015-01-01

    SUMMARY Information regarding the anatomy of the physiological apical foramen is limited. Knowing its diameter and shapes contributes to clinical work, specifically to the cleaning and shaping of the apical third. The aim of this ex vivo study was to determine the minimum and maximum diameters and shape of the physiological apical foramen in the roots of maxillary and mandibular first molars. A descriptive study was conducted on 89 recently extracted first molars. Roots 3–5 mm from the apex were sectioned and prepared for analysis at 40× magnification. The minimum and maximum diameters of each physiological foramen were measured using the program Motic Images plus 2.0 ML. The shape of the foramina, classified as round, oval or irregular, was determined by the difference between the maximum and minimum diameters. A total of 174 physiological foramina were analyzed. The average of the minimum and maximum diameters was between 0.24–0.33 mm in maxillary first molars and between 0.25–0.33 mm in mandibular first molars. In maxillary molars, the most common shape of the foramen was oval (50%), then irregular (32%), then round (18%). In mandibular molars, the oval shape was also the most frequent (59%), followed by irregular (23%) and round (18%). The findings of this study regarding the morphology of physiological apical foramina in first molars make it easier for the operator to choose the appropriately-sized instruments to perform endodontic therapy successfully. PMID:25937698

  2. Limited elimination of two viruses by cryotherapy of pelargonium apices related to virus distribution.

    Science.gov (United States)

    Gallard, A; Mallet, R; Chevalier, M; Grapin, A

    2011-01-01

    The possibility of eradicating the pelargonium flower break virus (PFBV) and pelargonium line pattern virus (PLPV) by cryotherapy of axillary shoot apices was investigated using five Pelargonium cultivars. Viruses were detected by DAS-ELISA and their location was determined by immunolocalization. Apex culture did not permit elimination of PFBV and only 15 percent regenerated plants of 'Stellar Artic' cultivar were ELISA PLPV-negative. Plants regenerated from cryotherapy-treated apices were tested by DAS-ELISA after a 3-month in vitro culture period. Viruses were not detected in 25 percent and 50 percent of the plants tested for PFBV and PLPV, respectively. However, immunolocalization carried out on apices originating from cryopreserved shoot tips sampled from DAS-ELISA negative plants showed that they were still virus-infected. Using immunolocalization, PFBV and PLPV could be detected in Pelargonium apices, even in the meristematic dome. However, viral particles were more numerous in basal zone cells than in meristematic cells. Our results demonstrate that PFBV and PLPV are present within meristematic cells and that cryopreservation can partly reduce the quantity of these viruses in Pelargonium plants but not eliminate them totally. Additional knowledge on localization and behaviour of viruses during cryopreservation is essential to optimize cryotherapy and plant genetic resource management.

  3. Assessment of the nonoperated root after apical surgery of the other root in mandibular molars

    DEFF Research Database (Denmark)

    Kraus, Riccardo D; von Arx, Thomas; Gfeller, David;

    2015-01-01

    INTRODUCTION: If a surgical approach is chosen to treat a multirooted tooth affected by persistent periapical pathosis, usually only the affected roots are operated on. The present study assessed the periapical status of the nonoperated root 5 years after apical surgery of the other root in mandi...

  4. Ultrasonic root-end preparation in apical surgery : a prospective randomized study

    NARCIS (Netherlands)

    de Lange, Jan; Putters, Thomas; Baas, Erik M.; van Ingen, Johan M.

    2007-01-01

    Objective. The purpose of this study was to evaluate the potential benefit of an ultrasonic device in apical surgery on the outcome of treatment. Study design. A randomized prospective design was used in a standardized treatment protocol. Patients were allocated to treatment with an ultrasonic devic

  5. Dual roles of Notch in regulation of apically restricted mitosis and apicobasal polarity of neuroepithelial cells.

    Science.gov (United States)

    Ohata, Shinya; Aoki, Ryo; Kinoshita, Shigeharu; Yamaguchi, Masahiro; Tsuruoka-Kinoshita, Sachiko; Tanaka, Hideomi; Wada, Hironori; Watabe, Shugo; Tsuboi, Takashi; Masai, Ichiro; Okamoto, Hitoshi

    2011-01-27

    How the mitosis of neuroepithelial stem cells is restricted to the apical ventricular area remains unclear. In zebrafish, the mosaic eyes(rw306) (moe/epb41l5(rw306)) mutation disrupts the interaction between the putative adaptor protein Moe and the apicobasal polarity regulator Crumbs (Crb), and impairs the maintenance of neuroepithelial apicobasal polarity. While Crb interacts directly with Notch and inhibits its activity, Moe reverses this inhibition. In the moe(rw306) hindbrain, Notch activity is significantly reduced, and the number of cells that proliferate basally away from the apical area is increased. Surprisingly, activation of Notch in the moe(rw306) mutant rescues not only the basally localized proliferation but also the aberrant neuroepithelial apicobasal polarity. We present evidence that the Crb⋅Moe complex and Notch play key roles in a positive feedback loop to maintain the apicobasal polarity and the apical-high basal-low gradient of Notch activity in neuroepithelial cells, both of which are essential for their apically restricted mitosis. PMID:21262462

  6. Cone beam computed tomography study of apical root resorption induced by Herbst appliance

    Science.gov (United States)

    SCHWARTZ, João Paulo; RAVELI, Taísa Boamorte; ALMEIDA, Kélei Cristina de Mathias; SCHWARTZ-FILHO, Humberto Osvaldo; RAVELI, Dirceu Barnabé

    2015-01-01

    Objective This study evaluated the frequency of root resorption during the orthodontic treatment with Herbst appliance by Cone Beam Computed Tomography (CBCT). Material and Methods The sample comprised 23 patients (11 men, 12 women; mean ages 15.76±1.75 years) with Class II division 1 malocclusion, treated with Herbst appliance. CBCT was obtained before treatment (T0) and after Herbst treatment (T1). All the dental roots, except third molars, were evaluated, and apical root resorption was determined using the axial guided navigation method. Paired t-tests and Wilcoxon T Test were used to compare the dependent samples in parametric and nonparametric cases, respectively. Chi-Square Test with Yates’ correction was used to evaluate the relationship between apical root resorption and gender. Results were considered at a significance level of 5%. Results Apical resorption was detected by CBCT in 57.96% of 980 roots that underwent Herbst appliance treatment. All patients had minimal resorption and there was no statistical significance between the genders. Conclusion CBCT three-dimensional evaluation showed association between Herbst appliance and minimal apical root resorption, mostly in the anchoring teeth, without clinical significance. PMID:26537718

  7. A systematic analysis of neonatal mouse heart regeneration after apical resection.

    Science.gov (United States)

    Bryant, Donald Marion; O'Meara, Caitlin Claire; Ho, Nhi Ngoc; Gannon, Joseph; Cai, Lei; Lee, Richard Theodore

    2015-02-01

    The finding that neonatal mice are able to regenerate myocardium after apical resection has recently been questioned. We determined if heart regeneration is influenced by the size of cardiac resection and whether surgical retraction of the ventricular apex results in an increase in cardiomyocyte cell cycle activity. We performed moderate or large apical ventricular resections on neonatal mice and quantified scar infiltration into the left ventricular wall at 21 days post-surgery. Moderately resected hearts had 15±2% of the wall infiltrated by a collagen scar; significantly greater scar infiltration (23±4%) was observed in hearts with large resections. Resected hearts had higher levels of cardiomyocyte cell cycle activity relative to sham hearts. Surgically retracting the ventricle often resulted in fibrosis and induced cardiomyocyte cell cycle activity that were comparable to that of resected hearts. We conclude that apical resection in neonatal mice induces cardiomyocyte cell cycle activity and neomyogenesis, although scarring can occur. Surgical technique and definition of approach to assessing the extent of regeneration are both critical when using the neonatal mouse apical resection model.

  8. Sugar demand, not auxin, is the initial regulator of apical dominance.

    Science.gov (United States)

    Mason, Michael G; Ross, John J; Babst, Benjamin A; Wienclaw, Brittany N; Beveridge, Christine A

    2014-04-22

    For almost a century the plant hormone auxin has been central to theories on apical dominance, whereby the growing shoot tip suppresses the growth of the axillary buds below. According to the classic model, the auxin indole-3-acetic acid is produced in the shoot tip and transported down the stem, where it inhibits bud growth. We report here that the initiation of bud growth after shoot tip loss cannot be dependent on apical auxin supply because we observe bud release up to 24 h before changes in auxin content in the adjacent stem. After the loss of the shoot tip, sugars are rapidly redistributed over large distances and accumulate in axillary buds within a timeframe that correlates with bud release. Moreover, artificially increasing sucrose levels in plants represses the expression of BRANCHED1 (BRC1), the key transcriptional regulator responsible for maintaining bud dormancy, and results in rapid bud release. An enhancement in sugar supply is both necessary and sufficient for suppressed buds to be released from apical dominance. Our data support a theory of apical dominance whereby the shoot tip's strong demand for sugars inhibits axillary bud outgrowth by limiting the amount of sugar translocated to those buds.

  9. Editing of CD1d-Bound Lipid Antigens by Endosomal Lipid Transfer Proteins

    OpenAIRE

    Zhou, Dapeng; Cantu, Carlos; Sagiv, Yuval; Schrantz, Nicolas; Kulkarni, Ashok B.; Qi, Xiaoyang; Mahuran, Don J.; Carlos R Morales; Grabowski, Gregory A.; Benlagha, Kamel; Savage, Paul; Bendelac, Albert; Teyton, Luc

    2003-01-01

    It is now established that CD1 molecules present lipid antigens to T cells, although it is not clear how the exchange of lipids between membrane compartments and the CD1 binding groove is assisted. We report that mice deficient in prosaposin, the precursor to a family of endosomal lipid transfer proteins (LTP), exhibit specific defects in CD1d-mediated antigen presentation and lack Vα14 NKT cells. In vitro, saposins extracted monomeric lipids from membranes and from CD1, thereby promoting the...

  10. Monoclonal antibodies to antigens on human neutrophils, activated T lymphocytes, and acute leukemia blast cells

    Energy Technology Data Exchange (ETDEWEB)

    Miterev, G.Yu.; Burova, G.F.; Puzhitskaya, M.S.; Danilevich, S.V.; Bulycheva, T.I.

    1987-11-01

    The authors describe the production of two mouse hybridomas secreting monoclonal antibodies to antigenic determinants of the surface membranes of human neutrophils, activated T lymphocytes, and acute leukemic blast cells. The degree of lymphocyte stimulation was estimated from incorporation of /sup 3/H-thymidine with parallel microculture. Monoclonal antibodies of supernatants of hybridoma cultures shown here reacted in both immunofluorescence test and cytotoxicity test with surface membrane antigens on the majority of neutrophils and PHA-activated peripheral blood lymphocytes from healthy subjects, but did not give positive reactions with unactivated lymphocytes, adherent monocytes, erythrocytes, and alloantigen-stimulated lymphocytes.

  11. Bioinformatic analysis for a new alternatively spliced variant of prostate-specific membrane antigen%前列腺特异性膜抗原新型剪接变异体PSME基因及蛋白的生物信息学分析

    Institute of Scientific and Technical Information of China (English)

    王铸; 冯发深; 何霞; 关林琳; 徐霖; 张定梅; 罗燕芬; 汪杨; 曹开源

    2012-01-01

    目的 通过对PSME的生物信息学分析,更多的了解该基因及蛋白结构与功能的相关信息.方法 运用生物信息学方法分析PSME基因结构,序列及其编码蛋白的理化性质和结构与功能特征,蛋白相互作用网络以及抗原表位.结果 用ExPASy的Computer PI/Mw、SMA RT等软件对其氨基酸序列进行分析,该蛋白的PI值为6.38,相对分子质量约为78 000.二级结构中α螺旋(H)占34.66%,β折叠(E)占12.93%,无规卷曲占52.41%.PSME信号肽位于1~22位氨基酸,150-248位为肽酶结构域,639~703位为转铁蛋白受体二聚体,且含有多个糖基化位点.通过DNA Star软件分析得到了PSME蛋白的抗原表位.结论 利用生物信息学预测出的结构和功能信息,能为PSME蛋白的相关研究提供信息基础.%This paper focuses on the prediction of structures and functions of a new alternatively spliced variant of prostate-specific membrane antigen (PSME) using bioinformatics methods. ExPASy's tools such as computer Pl/Mw, SMART and other software were used to analysis PSME amino acid sequence. Furthermore, protein interaction network was predicated by STRING, and B-cell epitopes of the protein was predicted with DNA Star. We fund that the protein PI value was 6.38, M, was 78 000, A-helix secondary structure (H) accounted for 34.66%, βfold (E) accounted for 12.93%, and random coil accounting for 5Z41%. PSME signal peptide was in the 1-22 bit, peptidase domain wan in 150-248, and TFR dimer in 639-703 bit. Furthermore, PSME was also contained multiple glycosylation sites. The predication of structures and functions lays down a information foundation for the further investigation of PSME.

  12. CASE OF DIAGNOSIS OF APICAL FORM OF HYPERTROPHIC CARDIOMYOPATHY WITH A PATIENT WITH PROGRESSIVE ANGINA CLINIC

    Directory of Open Access Journals (Sweden)

    N. S. Krylova

    2014-01-01

    Full Text Available Objective of work: to describe the apical form of hypertrophic cardiomyopathy (AFHC developing under the "mask" of the ischemic heart disease not diagnosed for a long period.Materials and methods. Patient B., 73 y.o., female, was brought to the cardiology department with complains of severe pressing pain behind the breastbone caused with no apparent reason and lasting for over 4 hours. The following examination of the patient was performed: electrocardiography (ECG, echocardiography (EchoCG, Holter ECG monitoring, coronary angiography (CAG, ventriculography.Results. The final diagnosis for the patient was set on the basis of the following readings: ECG data (basic rhythm – atrial fibrillation, left ventricle (LV hypertrophy, negative T-waves in leads V1–6, ST segment depression up to 1–2 mm in leads V4–6, EchoCG (hypertrophy of apical segments of the LV with decreasing of its cavity, moderate dilatation of the left atrium, intraventricular obstruction in the apical third of the LV with the maximum pressure gradient of up to 48 mm Hg., CAG (stenotic lesions of coronary arteries were found, ventriculography (LV volume is not increased, no violations of local contractility, narrowing of the LV cavity in the lower third is observed with thinning in the apex, which indicatesexpressed apical hypertrophy of the LV myocard. AFHC, apical form with moderate obstruction in the lower third of the left ventricle. Stress angina syndrome. CAG and ventriculography were main diagnostic methods that allowed setting the final diagnosis.Conclusion. The clinical case sets forth the peculiarities of diagnostics, therapy, and post-therapy management of patients with this form of AFHC.

  13. CASE OF DIAGNOSIS OF APICAL FORM OF HYPERTROPHIC CARDIOMYOPATHY WITH A PATIENT WITH PROGRESSIVE ANGINA CLINIC

    Directory of Open Access Journals (Sweden)

    N. S. Krylova

    2015-01-01

    Full Text Available Objective of work: to describe the apical form of hypertrophic cardiomyopathy (AFHC developing under the "mask" of the ischemic heart disease not diagnosed for a long period.Materials and methods. Patient B., 73 y.o., female, was brought to the cardiology department with complains of severe pressing pain behind the breastbone caused with no apparent reason and lasting for over 4 hours. The following examination of the patient was performed: electrocardiography (ECG, echocardiography (EchoCG, Holter ECG monitoring, coronary angiography (CAG, ventriculography.Results. The final diagnosis for the patient was set on the basis of the following readings: ECG data (basic rhythm – atrial fibrillation, left ventricle (LV hypertrophy, negative T-waves in leads V1–6, ST segment depression up to 1–2 mm in leads V4–6, EchoCG (hypertrophy of apical segments of the LV with decreasing of its cavity, moderate dilatation of the left atrium, intraventricular obstruction in the apical third of the LV with the maximum pressure gradient of up to 48 mm Hg., CAG (stenotic lesions of coronary arteries were found, ventriculography (LV volume is not increased, no violations of local contractility, narrowing of the LV cavity in the lower third is observed with thinning in the apex, which indicatesexpressed apical hypertrophy of the LV myocard. AFHC, apical form with moderate obstruction in the lower third of the left ventricle. Stress angina syndrome. CAG and ventriculography were main diagnostic methods that allowed setting the final diagnosis.Conclusion. The clinical case sets forth the peculiarities of diagnostics, therapy, and post-therapy management of patients with this form of AFHC.

  14. Apical crown technique to model canal roots. A review of the literature.

    Science.gov (United States)

    Tortini, D; Colombo, M; Gagliani, M

    2007-09-01

    The goals of endodontic therapy are to shape and clean the root canal system in order to receive a three-dimensional and hermetic filling of the entire endodontic space. The aim of this review is to analyze the development of the different canal shaping techniques from the manual stainless-steel instruments to the recently developed rotary Ni-Ti systems. The need to provide instruments of a suitable size to reach the apex of curved root canals drove the development of preparation techniques with a greater focus on the apical part. The Crown-Down technique involves the widening of the canal orifices with Gates-Glidden drills followed by the incremental removal of organic canal contents proceeding from the canal orifice to the apical portion using manual files. Files are used from larger to smaller with no apical pressure. Nevertheless, the term ''Crown-Down'' does not define the technique in the strictest sense, nor does it provide a specific sequence for the use of instruments, but rather defines a way to use the instruments. The preparation of root canals has been revolutionised by the development of Ni-Ti alloys. The features of this alloy give instruments profound flexibility and allow the manufacture of more tapered instruments compared with traditional stainless-steel instruments. In the second part of this review, numerous examples are provided concerning the comparison between different Ni-Ti systems and their techniques for use. A Crown-Down approach provides certain advantages including early organic debris removal, the creation of a large reservoir for irrigating solutions, a straighter access to the apical region of curved canals, and greater precision with regard to the exact working length and apical size.

  15. Morphology of bacterial flora in root canals associated with apical abscesses

    Institute of Scientific and Technical Information of China (English)

    Guo Huijie; Gao Chengzhi; Zhang Chengfei; Zheng Shuying; Yue Lin

    2014-01-01

    Background Apical abscess is an inflammatory process in the peri-radicular tissues caused by biofllms in the necrotic root canal systems.Therefore,a comprehensive analysis of the bacterial colonization is required for a better understanding of the pathogenesis.This study aimed to investigate the patterns of bacterial infection of root canals of teeth with apical abscesses and to determine whether histological and microbiological findings correlated with clinical conditions.Methods Eighteen samples from 18 teeth with apical pathological lesions were analyzed.Nine patients with acute apical abscesses experienced severe pain,and nine patients were asymptomatic with a sinus tract.After extraction,each affected root was divided into two halves.One half was processed for histobacteriologic analysis and examined using light microscopy,and the other half was analyzed using scanning electron microscopy (SEM) to determine the patterns of microbial colonization of the root canals.Results The appearance of each sample subjected to SEM was consistent with the histobacteriologic findings despite the presence or absence of clinical symptoms.Intraradicular biofilms comprising cocci,rods,and/or filaments of amorphous materials were observed in the apical third of the main root canals in all samples.The bacterial biofilms covering the main root canal walls also penetrated the dentinal tubules to varying depths.The morphologies of biofilms varied,and a unique pattern of intraradicular infection was not identified.Conclusion Intraradicular infections formed complex and variable multispecies biofilms and their presence did not correlate with clinical symptoms.

  16. Evaluation of the endodontic apical seal after post insertion by synchrotron radiation microtomography

    International Nuclear Information System (INIS)

    The commonly used methods for evaluating the endodontic apical seal, such as longitudinal and transversal section and diaphanization, show some operative difficulties and intrinsic limitation. This study suggests and describes a new method of analysis using a synchrotron radiation microtomography to analyse the root apex after post insertion, creating a three-dimensional image and analysing sections of the specimen every 5μm. The study was performed at SYRMEP beam line at the Electra Synchrotron in Trieste using monochromatic X-rays of 32KeV. Eleven monoradicular teeth were prepared using NiTi GT Rotary files instruments to an apical size 20 with conicity .06 and divided in four groups: in G1 (n=4) and G2 (n=2), the specimens were endodontically filled with guttapercha and a zinc-oxide sealer, in G3 (n=3) and G4 (n=2) guttapercha and a silicon-based sealer were used. An endodontic post was inserted in specimens of groups 1 and 3 following the manufacturer's instructions. Specimens were analysed using monochromatic X-rays of 32KeV. A CCD detector with pixel dimension pf 5x5μm2 was used for the acquisition process. Seven hundred and twenty projections were performed over 180o range using a high-resolution rotator. The projections were reconstructed using standard algorithms for tomographic reconstruction. The apical infiltration was evaluated by verifying if black spots were detectable on the images. The specimens of groups 3 and 4 showed a better apical seal than the ones of groups 1 and 2. Post insertion, when a ZOE-based sealer is used, increases the apical gap even if it does not seem to be clinically relevant and sufficient to be a cause of endodontic failure. The new method for analysis appeared to be effective to detect small defects in endodontic obturation, analysing guttapercha-sealer and sealer-dentin interfaces

  17. Uterine receptivity and the plasma membrane transformation

    Institute of Scientific and Technical Information of China (English)

    Christopher R MURPHY

    2004-01-01

    This review begins with a brief commentary on the diversity of placentation mechanisms, and then goes on to examine the extensive alterations which occur in the plasma membrane of uterine epithelial cells during early pregnancy across species. Ultrastructural, biochemical and more general morphological data reveal that strikingly common phenomena occur in this plasma membrane during early pregnancy despite the diversity of placental types-from epitheliochorial to hemochorial, which ultimately form in different species. To encapsulate the concept that common morphological and molecular alterations occur across species, that they are found basolaterally as well as apically, and that moreover they are an ongoing process during much of early pregnancy, not just an event at the time attachment,brane during early pregnancy are key to uterine receptivity.

  18. Release of carcinoembryonic antigen from human colon cancer cells by phosphatidylinositol-specific phospholipase C.

    OpenAIRE

    Sack, T L; Gum, J R; Low, M G; Y. S. Kim

    1988-01-01

    Carcinoembryonic antigen (CEA) is released from colon cancer cells into the circulation where it is monitored clinically as an indicator of the recurrence or progression of cancer. We have studied the mechanism of CEA membrane attachment and release using the human colonic adenocarcinoma cell line LS-174T, specimens of human colon cancers, and serum from colon cancer patients. CEA release by cells in vitro and in vivo is associated with the conversion of CEA from a membrane-bound, hydrophobic...

  19. Immunohistochemical features of the granulation tissue structure in the primary and secondary apical periodontitis

    Directory of Open Access Journals (Sweden)

    Gritsenko P.I.

    2015-03-01

    Full Text Available Background The search of criteria of differential enzymatic activity of cyclooxygenase-2 and matrix metalloproteinases immunocompetent cells of granulomas in the primary and secondary apical periodontitis (AP provides information about the features of lesions of periodontal stromal component, and it can be used as the basis for future therapeutic intervention. Objective. The purpose of this study was to compare the qualitative and quantitative characteristics of the cellular composition of granulation tissue, which formed during primary and secondary apical periodontitis; comparison of morphological characteristics of the structure apical granuloma with immunohistochemical indicators of markers expression of cluster of differentiation CD3, CD4, CD8, CD68, CD138, CD20 and cyclooxygenase-2 and matrix metalloproteinase 9. Methods. The study included 59 patients aged 18 to 50 years with apical periodontitis. Patients were divided into 2 clinical group: 1 – with primary periodontitis and 2 – with secondary periodontitis. For morphological studies using peryapikalnoyi granulation tissue biopsies. Results. Significant differences were found for the fractions of CD4+ T helper cells (p=0,034, r=0,321, CD20+ mature B lymphocytes (p=0,001, r=0,671, CD68+ macrophages (p=0,011, r=0,684 and CD138+ plasma cells (p=0,002, r=0,645. And, also likely decrease in the ratio of CD4+/CD8+ (p=0,002, r=0,645, by reducing the fraction of CD4+ T-helper cells, against reducing the number of leukocytes, expression of MMP-9 (p=0,04, r=0,302 and COX-2 (p=0,013, r=0,321 with secondary apical periodontitis, compared to the primary periodontitis. Conclusion. Disease progression and the beginning of the destruction of bone tissue with the formation of granulation tissue is the result of abnormal immune responses, III and IV type hypersensitivity reactions, with the formation of immune granulomas. Citation: Gritsenko PI, Samojlenko AV, Shpon’ka IS, Poslavska

  20. Identification of Schistosoma mansoni candidate antigens for diagnosis of schistosomiasis

    Directory of Open Access Journals (Sweden)

    Gardenia Braz Figueiredo Carvalho

    2011-11-01

    Full Text Available The development of a more sensitive diagnostic test for schistosomiasis is needed to overcome the limitations of the use of stool examination in low endemic areas. Using parasite antigens in enzyme linked immunosorbent assay is a promising strategy, however a more rational selection of parasite antigens is necessary. In this study we performed in silico analysis of the Schistosoma mansoni genome, using SchistoDB database and bioinformatic tools for screening immunogenic antigens. Based on evidence of expression in all parasite life stage within the definitive host, extracellular or plasmatic membrane localization, low similarity to human and other helminthic proteins and presence of predicted B cell epitopes, six candidates were selected: a glycosylphosphatidylinositol-anchored 200 kDa protein, two putative cytochrome oxidase subunits, two expressed proteins and one hypothetical protein. The recognition in unidimensional and bidimensional Western blot of protein with similar molecular weight and isoelectric point to the selected antigens by sera from S. mansoni infected mice indicate a good correlation between these two approaches in selecting immunogenic proteins.

  1. Adenovirus entry from the apical surface of polarized epithelia is facilitated by the host innate immune response.

    Directory of Open Access Journals (Sweden)

    Poornima L N Kotha

    2015-03-01

    Full Text Available Prevention of viral-induced respiratory disease begins with an understanding of the factors that increase or decrease susceptibility to viral infection. The primary receptor for most adenoviruses is the coxsackievirus and adenovirus receptor (CAR, a cell-cell adhesion protein normally localized at the basolateral surface of polarized epithelia and involved in neutrophil transepithelial migration. Recently, an alternate isoform of CAR, CAREx8, has been identified at the apical surface of polarized airway epithelia and is implicated in viral infection from the apical surface. We hypothesized that the endogenous role of CAREx8 may be to facilitate host innate immunity. We show that IL-8, a proinflammatory cytokine and a neutrophil chemoattractant, stimulates the protein expression and apical localization of CAREx8 via activation of AKT/S6K and inhibition of GSK3β. Apical CAREx8 tethers infiltrating neutrophils at the apical surface of a polarized epithelium. Moreover, neutrophils present on the apical-epithelial surface enhance adenovirus entry into the epithelium. These findings suggest that adenovirus evolved to co-opt an innate immune response pathway that stimulates the expression of its primary receptor, apical CAREx8, to allow the initial infection the intact epithelium. In addition, CAREx8 is a new target for the development of novel therapeutics for both respiratory inflammatory disease and adenoviral infection.

  2. Membrane targeting of cGMP-dependent protein kinase is required for cystic fibrosis transmembrane conductance regulator Cl- channel activation

    NARCIS (Netherlands)

    A.B. Vaandrager (Arie); A. Smolenski; B.C. Tilly (Bernard); A.B. Houtsmuller (Adriaan); E.M.E. Ehlert (Ehrich); A.G. Bot (Alice); M.J. Edixhoven (Marcel); W.E. Boomaars (Wendy); S.M. Lohmann (Suzanne); H.R. de Jonge (Hugo)

    1998-01-01

    textabstractA recently cloned isoform of cGMP-dependent protein kinase (cGK), designated type II, was implicated as the mediator of cGMP-provoked intestinal Cl- secretion based on its localization in the apical membrane of enterocytes and on its capacity to activate cys

  3. Cdc42-dependent Modulation of Tight Junctions and Membrane Protein Traffic in Polarized Madin-Darby Canine Kidney Cells

    Science.gov (United States)

    Rojas, Raul; Ruiz, Wily G.; Leung, Som-Ming; Jou, Tzuu-Shuh; Apodaca, Gerard

    2001-01-01

    Polarized epithelial cells maintain the asymmetric composition of their apical and basolateral membrane domains by at least two different processes. These include the regulated trafficking of macromolecules from the biosynthetic and endocytic pathway to the appropriate membrane domain and the ability of the tight junction to prevent free mixing of membrane domain-specific proteins and lipids. Cdc42, a Rho family GTPase, is known to govern cellular polarity and membrane traffic in several cell types. We examined whether this protein regulated tight junction function in Madin-Darby canine kidney cells and pathways that direct proteins to the apical and basolateral surface of these cells. We used Madin-Darby canine kidney cells that expressed dominant-active or dominant-negative mutants of Cdc42 under the control of a tetracycline-repressible system. Here we report that expression of dominant-active Cdc42V12 or dominant-negative Cdc42N17 altered tight junction function. Expression of Cdc42V12 slowed endocytic and biosynthetic traffic, and expression of Cdc42N17 slowed apical endocytosis and basolateral to apical transcytosis but stimulated biosynthetic traffic. These results indicate that Cdc42 may modulate multiple cellular pathways required for the maintenance of epithelial cell polarity. PMID:11514615

  4. Structure of the major membrane protein complex from urinary bladder epithelial cells by cryo-electron crystallography

    NARCIS (Netherlands)

    Oostergetel, GT; Keegstra, W; Brisson, A

    2001-01-01

    Numerous protein plaques cover the apical surface of mammalian urinary bladder epithelial cells. These plaques contain four integral membrane proteins, called uroplakins, which form a well-ordered array of hexameric complexes. The 3D structure of these naturally occurring 2D crystals was studied by

  5. Selection of protective antigens in Lawsonia intracellularis by reverse vaccinology

    DEFF Research Database (Denmark)

    Vadekær, Dorte Fink; Lundegaard, Claus; Riber, Ulla;

    in Denmark. Experimental challenge studies previously performed at DTU-Vet show that a primary infection results in complete protection against reinfection due to induction of immunological memory. We aim to develop a subunit vaccine that mimics the induction of the immune response and hence causes...... protection against L. intracellularis. To this end, a reverse vaccinology approach was applied: the entire L. intracellularis genome encoding 1340 proteins was screened in silico using bioinformatics tools to identify potential protein antigens. Advanced software algorithms predicted 150 secreted and outer...... membrane proteins, and these were analyzed and given a score for presence of B and T cell epitopes. Using another in silico technology platform, which identifies novel B cell antigens eliciting a highly protective immune response, we obtained a second list of potential vaccine candidates. Six proteins were...

  6. Filtración apical versus filtración coronal

    OpenAIRE

    Tabares Martínez, Pablo

    2016-01-01

    La filtración del conducto radicular es la principal causa de los fracasos en endodoncia (19). Esta recontaminación puede producirse a través de dos vías principales de filtración que son los orificios coronal y apical. En la actualidad hay controversia entre diferentes autores sobre cuál es la vía de filtración más importante, si la apical (9, 167) o la coronal (4, 76). Existen diversas técnicas de obturación y materiales de obturación, entre las que puede elegir el clínico en función de cad...

  7. Micro‐CT analyses of apical enlargement and molar root canal complexity

    DEFF Research Database (Denmark)

    Markvart, M.; Darvann, Tron Andre; Larsen, P.;

    2012-01-01

    Markvart M, Darvann TA, Larsen P, Dalstra M, Kreiborg S, Bjørndal L. Micro‐CT analyses of apical enlargement and molar root canal complexity. International Endodontic Journal, 45, 273–281, 2012. Aim To compare the effectiveness of two rotary hybrid instrumentation techniques with focus on apical...... enlargement in molar teeth and to quantify and visualize spatial details of instrumentation efficacy in root canals of different complexity. Methodology Maxillary and mandibular molar teeth were scanned using X‐ray microcomputed tomography. Root canals were prepared using either a GT/Profile protocol or a Ra......Ce/NiTi protocol. Variables used for evaluation were the following: distance between root canal surfaces before and after preparation (distance after preparation, DAP), percentage of root canal area remaining unprepared and increase in canal volume after preparation. Root canals were classified according to size...

  8. Inadvertent Apical Extrusion of Sodium Hypochlorite with Evaluation by Dental Volumetric Tomography

    Directory of Open Access Journals (Sweden)

    Elif Delve Başer Can

    2015-01-01

    Full Text Available This case report describes the tissue injury caused by inadvertently extruded NaOCl through the apical constriction. A 56-year-old female patient with complaints of pain, swelling, and ecchymosis on the left side of her face was referred to our clinic. The symptoms had emerged following root canal treatment of the maxillary left first premolar, and a soft tissue complication due to apical extrusion of NaOCl was diagnosed. Antibiotics and analgesics were prescribed. DVT images revealed that the buccal root apex had perforated the maxillary bone. The patient was followed up every other day and became asymptomatic on the 10th day. Endodontic therapy was completed with routine procedures. Determining working length precisely and following irrigation protocols meticulously are indispensable to prevent this type of complication. 3D visualization of the affected area may reveal the cause of the incident.

  9. [Endodontic re-treatment or apical surgery, is an evidence-based choice possible?].

    Science.gov (United States)

    de Lange, J

    2016-02-01

    Treatment of a recurrence after an initial endodontic treatment can consist of endodontic re-treatment or apical surgery. The literature reports comparable success rates for these two options. However, randomised controlled trials that are truly comparable to each other are unavailable as a result of which comparison and an informed choice are not really possible. In addition to the treatment outcome, consideration also has to be given to cost-effectiveness and other patient-related factors, such as pain, swelling and medication. Regarding these considerations, too, little reliable information can be found in the literature. Moreover, factors relating to insurance play an important role in the Netherlands, where only apical surgery is covered by the basic healthcare insurance. In conclusion, all of these factors should be carefully considered before proceeding to an individual treatment strategy. PMID:26878715

  10. Multiple Apical Radiolucencies and External Cervical Resorption Associated with Varicella Zoster Virus: A Case Report.

    Science.gov (United States)

    Patel, Kreena; Schirru, Elia; Niazi, Sadia; Mitchell, Philip; Mannocci, Francesco

    2016-06-01

    Varicella zoster virus (VZV) is responsible for the primary infection chickenpox. After the initial infection, it remains latent but can reactivate, resulting in shingles (herpes zoster). Previous reports have implicated VZV in the pathogenesis of apical periodontitis, but the involvement of the virus has not been investigated fully. The present case describes a patient who suffered from a severe episode of shingles and subsequently developed periapical radiolucencies of all the teeth in the affected nerve distribution. Molecular and culture techniques showed the presence of VZV DNA in the root canal system in the absence of bacteria. This confirms that VZV can cause localized pulp necrosis and apical periodontitis. The lesions healed after endodontic treatment, implying chemomechanical debridement using sodium hypochlorite irrigation and a calcium hydroxide interim dressing may be effective against the virus. PMID:27133503

  11. Neurons derive from the more apical daughter in asymmetric divisions in the zebrafish neural tube.

    Science.gov (United States)

    Alexandre, Paula; Reugels, Alexander M; Barker, David; Blanc, Eric; Clarke, Jonathan D W

    2010-06-01

    In the developing CNS, asymmetric cell division is critical for maintaining the balanced production of differentiating neurons while renewing the population of neural progenitors. In invertebrates, this process depends on asymmetric inheritance of fate determinants during progenitor divisions. A similar mechanism is widely believed to underlie asymmetrically fated divisions in vertebrates, but compelling evidence for this is missing. We used live imaging of individual progenitors in the intact zebrafish embryo CNS to test this hypothesis. We found that asymmetric inheritance of a subcellular domain is strongly correlated with asymmetric daughter fates and our results reveal an unexpected feature of this process. The daughter cell destined to become a neuron was derived from the more apical of the two daughters, whereas the more basal daughter inherited the basal process and replenished the apical progenitor pool.

  12. Apical ballooning with mid-ventricular obstruction: the many faces of Takotsubo cardiomyopathy.

    Science.gov (United States)

    Spadotto, Veronica; Elmaghawry, Mohamed; Zorzi, Alessandro; Migliore, Federico; Marra, Martina Perazzolo

    2013-01-01

    Takotsubo cardiomyopathy (TTC) is a transient left ventricular dysfunction due to akinesia of the left-ventricular (LV) mid-apical segments (apical ballooning), which can cause severe reduction in LV systolic function. The typical clinical picture of TTC include chest pain, electrocardiographic changes consisting of mild ST-segment elevation followed by diffuse deep T-wave inversion, QTc interval prolongation and mild troponin release in the absence of significant coronary stenoses. The syndrome often affects post-menopausal women and is triggered by sympathetic overstimulation, like intense physical or emotional stress, so that it is called the "broken heart syndrome". Although left-ventricular systolic dysfunction usually fully recovers within few days, heart failure can still complicate the early phase. We report a case of stress-induced cardiomyopathy that had full recovery after 4 weeks of follow up. The main electrocardiographic, angiographic and imaging features are discussed. PMID:24689016

  13. The effect of root preparation technique and instrumentation length on the development of apical root cracks.

    Science.gov (United States)

    Adorno, Carlos G; Yoshioka, Takatomo; Suda, Hideaki

    2009-03-01

    The purpose of this study was to compare the effects of root canal preparation techniques and instrumentation length on the development of apical root cracks. Forty extracted mandibular premolars with straight roots were randomly selected and mounted on resin blocks with simulated periodontal ligaments, and the apex was exposed. The teeth were divided into four groups of 10 teeth each for different canal preparation techniques and instrumentation lengths: group A: step-back preparation (SB) with stainless steel files (SF) using root canal length (RCL) to guide instrumentation length; group B: SB using RCL - 1 mm; group C: crown-down preparation (CD) with Profile using RCL; and group D: CD with PF using RCL - 1 mm. Digital images of the instrumentation sequence were compared for each tooth. Statistical analysis revealed a significant effect of instrumentation length (p 0.05) on the development of apical cracks.

  14. [Farmer's lung antigens in Germany].

    Science.gov (United States)

    Sennekamp, J; Joest, M; Sander, I; Engelhart, S; Raulf-Heimsoth, M

    2012-05-01

    Recent studies suggest that besides the long-known farmer's lung antigen sources Saccharopolyspora rectivirgula (Micropolyspora faeni), Thermoactinomyces vulgaris, and Aspergillus fumigatus, additionally the mold Absidia (Lichtheimia) corymbifera as well as the bacteria Erwinia herbicola (Pantoea agglomerans) and Streptomyces albus may cause farmer's lung in Germany. In this study the sera of 64 farmers with a suspicion of farmer's lung were examined for the following further antigens: Wallemia sebi, Cladosporium herbarum, Aspergillus versicolor, and Eurotium amstelodami. Our results indicate that these molds are not frequent causes of farmer's lung in Germany. PMID:22477566

  15. One-step apexification using platelet rich fibrin matrix and mineral trioxide aggregate apical barrier

    Directory of Open Access Journals (Sweden)

    Anisha Kumar

    2014-01-01

    In one-step apexification using MTA, the technical problem encountered is controlling the overfill or underfill of MTA. The use of a matrix material helps to overcome this shortcoming. Platelet rich fibrin (PRF is an immune platelet concentrate, which can be used as a matrix, it also promotes wound healing and repair. This case report presents a case of one step apexification using MTA as an apical barrier and autologous PRF as an internal matrix.

  16. Uptake and Distribution of Aluminum in Root Apices of Two Rice Varieties under Aluminum Stress

    Directory of Open Access Journals (Sweden)

    MIFTAHUDIN

    2007-09-01

    Full Text Available Aluminum (Al toxicity is the major limiting factor of plant growth and production in acid soils. The target of Al toxicity is the root tip, which affects mainly on root growth inhibition. The aim of this research was to study the uptake and distribution of Al in root apices of two rice varieties IR64 (Al-sensitive and Krowal (Al-tolerant, which were grown on nutrient solution containing 0, 15, 30, 45, and 60 ppm of Al. The root growth was significantly inhibited in both rice varieties at as low as 15 ppm Al concentration. The adventive roots of both varieties showed stunted growth in respons to Al stress. There was no difference in root growth inhibition between both rice varieties as well as among Al concentrations. Al uptake on root apices was qualitatively and quantitatively analyzed. Histochemical staining of roots using hematoxylin showed dark purple color on 1 mm region of Al-treated root apices. Rice var. IR 64 tended to take up more Al in root tip than Krowal did. However, there was no statistically significant difference (p = 0.176 in root Al content of both varieties in response to different concentration and period of Al treatments. Al distribution in root apices was found in the epidermal and subepidermal region in both rice varieties. Based on those results, rice var. Krowal that was previously grouped as Al-tolerant variety has similar root growth and physiological response to Al stress as compared to Al-sensitive variety IR64.

  17. Effects of Long-term Right Ventricular Apical Pacing on Left Ventricular Remodeling and Cardiac Function

    Institute of Scientific and Technical Information of China (English)

    2009-01-01

    Objective: To investigate the impacts of long-term right ventricular apical pacing on the ventricular remodeling and cardiac functions of patients with high-grade and third-degree atrioventricular blockage with normal heart structures and cardiac functions. In addition, we provide many evidences for choosing an optimal electrode implantation site.Methods: Study participants included patients who were admitted for pacemaker replacements and revisited for examinations of implanted pacemakers at outpatient. Pa...

  18. Apical groove type and molecular phylogeny suggests reclassification of Cochlodinium geminatum as Polykrikos geminatum.

    Directory of Open Access Journals (Sweden)

    Dajun Qiu

    Full Text Available Traditionally Cocholodinium and Gymnodinium sensu lato clade are distinguished based on the cingulum turn number, which has been increasingly recognized to be inadequate for Gymnodiniales genus classification. This has been improved by the combination of the apical groove characteristics and molecular phylogeny, which has led to the erection of several new genera (Takayama, Akashiwo, Karenia, and Karlodinium. Taking the apical groove characteristics and molecular phylogeny combined approach, we reexamined the historically taxonomically uncertain species Cochlodinium geminatum that formed massive blooms in Pearl River Estuary, China, in recent years. Samples were collected from a bloom in 2011 for morphological, characteristic pigment, and molecular analyses. We found that the cingulum in this species wraps around the cell body about 1.2 turns on average but can appear under the light microscopy to be >1.5 turns after the cells have been preserved. The shape of its apical groove, however, was stably an open-ended anticlockwise loop of kidney bean shape, similar to that of Polykrikos. Furthermore, the molecular phylogenetic analysis using 18S rRNA-ITS-28S rRNA gene cistron we obtained in this study also consistently placed this species closest to Polykrikos within the Gymnodinium sensu stricto clade and set it far separated from the clade of Cochlodinium. These results suggest that this species should be transferred to Polykrikos as Polykrikos geminatum. Our results reiterate the need to use the combination of apical groove morphology and molecular phylogeny for the classification of species within the genus of Cochlodinium and other Gymnodiniales lineages.

  19. Coordinated regulation of apical hook development by gibberellins and ethylene in etiolated Arabidopsis seedlings

    Institute of Scientific and Technical Information of China (English)

    Fengying An; Xing Zhang; Ziqiang Zhu; Yusi Ji; Wenrong He; Zhiqiang Jiang; Mingzhe Li; Hongwei Guo

    2012-01-01

    Dark-grown Arabidopsis seedlings develop an apical hook when germinating in soil,which protects the cotyledons and apical meristematic tissues when protruding through the soil.Several hormones are reported to distinctly modulate this process.Previous studies have shown that ethylene and gibberellins (GAs) coordinately regulate the hook development,although the underlying molecular mechanism is largely unknown.Here we showed that GA3 enhanced while paclobutrazol repressed ethylene- and EIN3-overexpression (EIN3ox)-induced hook curvature,and della mutant exhibited exaggerated hook curvature,which required an intact ethylene signaling pathway.Genetic study revealed that GA-enhanced hook development was dependent on HOOKLESS 1 (HLS1),a central regulator mediating the input of the multiple signaling pathways during apical hook development.We further found that GA3 induced (and DELLA proteins repressed) HLS1 expression in an ETHYLENE INSENSITIVE 3/EIN3-LIKE 1 (EIN3/EIL1)-dependent manner,whereby EIN3/EIL1 activated HLS1 transcription by directly binding to its promoter.Additionally,DELLA proteins were found to interact with the DNA-binding domains of EIN3/EIL1 and repress EIN3/EIL1-regulated HLS1 expression.Treatment with naphthylphthalamic acid,a polar auxin transport inhibitor,repressed the constitutively exaggerated hook curvature of EIN3ox line and della mutant,supporting that auxin functions downstream of the ethylene and GA pathways in hook development.Taken together,our results identify EIN3/EIL1 as a new class of DELLA-associated transcription factors and demonstrate that GA promotes apical hook formation in cooperation with ethylene partly by inducing the expression of HLS1 via derepression of EIN3/EIL1 functions.

  20. ABA mediates PEG-mediated premature differentiation of root apical meristem in plants

    OpenAIRE

    Ji, Hongtao; Li, Xia

    2014-01-01

    Root apical meristem (RAM) is central for indeterminate growth of plant roots and in sensing environmental stimuli, such as water status. Recently, we reported that PEG8000-simulated mild and moderate osmotic stress induces premature differentiation of RAM, which is a conserved adaptive mechanism in higher plants to cope with water stress. Microarray data analysis revealed that the ABA signaling pathway may be involved in water stress-induced RAM premature differentiation. Here we showed that...

  1. Evaluation of the anatomical alterations of lower molars mesial root?s apical third

    Directory of Open Access Journals (Sweden)

    FRÖNER Izabel Cristina

    1999-01-01

    Full Text Available The anatomical apex of the mesial root of the lower molars presents a morphological complexity related to the number and shape of the root canals as well as of the apical foramen and isthmus presence. The knowledge of the complexity of the endodontic system of the molar root area is essencial to select more carefully the best instrumentation and obturation technique, to obtain a more successful endodontic therapy.

  2. A new approach to supply chain management based on pooling ITIL and APICS Principles and Practices

    Directory of Open Access Journals (Sweden)

    Abdelaali Himi

    2011-07-01

    Full Text Available APICS (Advanced Productivity, Innovation and Competitive Success - The Association for Operations Management is known as the repository providing a set of best practices in Supply Chain Management. The ITIL (Information Technology Infrastructure Library also provides best practices for IT service management. Noting the existence of a similarity between these two standards, this paper identifies the ITIL concepts used in the supply chain context and describe their contributions.

  3. Comparing the apical microleakage of lateral condensation and chloroform dip techniques with a new obturation method

    Directory of Open Access Journals (Sweden)

    Masoud Saatchi

    2011-01-01

    Full Text Available Background: The final objective of root canal therapy is to create a hermetic seal along the length of the root canal system. For this purpose, many methods and materials have been introduced. The purpose of this study was to compare the apical microleakage in a new obturation technique (true-tug-back with two other obturation techniques (lateral condensation and chloroform dip technique. Methods: In this in vitro study 102 single canal teeth were selected. The crowns were removed, and the canals were prepared using step-back technique. The master apical file was K-file #40. The teeth were divided into 3 experimental groups of 32 teeth. First group were obturated with lateral condensation technique and second group with chloroform dip technique and the third group with true-tug-back technique. Six teeth were used as control group. The teeth were placed in incubator at 100% humidity and 37°c for three days. The roots of the teeth were coated with two layers of nail varnish except for the apical 2 millimeter. Teeth were placed in Methylene blue 2% for one week. The teeth were sectioned vertically and the depth of maximum dye penetration for each tooth was recorded by stereomicroscope. Data were analyzed using ANOVA and Dunkan test. Results: The mean liner dye penetration differences between lateral condensation group (6.88 ± 4.06 mm and chloroform dip technique group (7.16 ± 3.37 mm were not statistically significant (P = 0.719. The differences between true-tug-back group (3.15 ± 0.52 mm and two other groups were statistically significant (P < 0.001. Conclusion: The results of this study showed that the true-tug-back technique can improve apical seal. Further studies are needed for this purpose.

  4. Pseudo-acute myocardial infarction due to transient apical ventricular dysfunction syndrome (Takotsubo syndrome)

    OpenAIRE

    Maciel, Bruno Araújo; Cidrão, Alan Alves de Lima; Sousa, Ítalo Bruno dos Santos; Ferreira, José Adailson da Silva; Messias Neto, Valdevino Pedro

    2013-01-01

    Takotsubo syndrome is characterized by predominantly medial-apical transient left ventricular dysfunction, which is typically triggered by physical or emotional stress. The present article reports the case of a 61-year-old female patient presenting with dizziness, excessive sweating, and sudden state of ill feeling following an episode involving intense emotional stress. The physical examination and electrocardiogram were normal upon admission, but the troponin I and creatine kinase-MB concen...

  5. 24 hour ST segment analysis in transient left ventricular apical ballooning.

    Directory of Open Access Journals (Sweden)

    Frank Bode

    Full Text Available OBJECTIVE: The etiologic basis of transient left ventricular apical ballooning, a novel cardiac syndrome, is not clear. Among the proposed pathomechanisms is coronary vasospasm. Long-term ST segment analysis may detect vasospastic episodes but has not been reported. METHODS: 30 consecutive patients with transient left ventricular apical ballooning, left ventricular dysfunction and normal or near-normal coronary arteries were investigated. A 24-hour Holter ECG was obtained after emergency admission. ST segment analysis was performed automatically in 2 leads and confirmed by visual inspection. Criteria for an ischemic event were: 1. ST elevation or 2. horizontal or down-sloping ST segments ≥1 min duration and ≥100 µV J+80 point deviation corrected for baseline ST-deviation. RESULTS: Patients presented with ST segment elevation (n = 19 and/or T wave inversion (n = 20 on admission ECG. Ejection fraction was 50±12%. No transient ST elevations were observed during Holter ECG analysis. In 3 patients, 8 transient episodes of ST depression were recorded. Durations of episodes varied between 75 s and 790 s (mean 229 s. Maximal ST deviation averaged -191±71 µV. Ischemic burden was -1 to -22 mVs (mean -8 mVs. 27 patients showed no ischemic events. CONCLUSIONS: ST segment analysis of 24 h Holter recordings revealed minor ischemic events in only 10% of patients with transient left ventricular apical ballooning. Overall, ST segment changes were not indicative of recurrent coronary spasm playing a major role in the genesis of transient left ventricular apical ballooning.

  6. Assessment of apically extruded debris and irrigant produced by different nickel-titanium instrument systems

    Directory of Open Access Journals (Sweden)

    Ebru KÜÇÜKYILMAZ

    2015-01-01

    Full Text Available The aim of this study was to evaluate the preparation time and the amounts of apically extruded debris and irrigant using different nickel-titanium instrumentation systems. Forty-five extracted single-rooted mandibular premolar teeth were selected and divided into three groups. The root canals were instrumented according to the manufacturers’ instructions, using a reciprocating single-file system, a single-file rotary system and a multiple-file rotary system. Bidistilled water was used as the irrigant solution. The apically extruded debris and irrigant were collected into preweighed Eppendorf tubes. The amounts of extruded debris and irrigant were assessed with a precision micro-balance. The Eppendorf tubes were incubated at 37°C for 15 days. After the incubation period, they were weighed again to assess the debris extrusion. The time required to prepare the canals was also recorded. The results were statistically analyzed using MANOVA and Bonferroni’s adjustment. Considering the apically extruded debris and irrigant, there were no statistically significant differences among the groups (p > 0.05. The Reciproc group produced the highest debris (0.000632 ± 0.000162 gr and irrigant (0.844587 ± 0.437814 ml extrusion values. While the least extruded debris was observed with OneShape (0,000431 ± 0,000171 gr, the least extruded irrigant was observed with ProTaper system (0.564147 ± 0.370596 ml. Instrumentation was faster using the Reciproc than the other two instruments (70.27 ± 13.38 s (p < 0.05. All of the instrumentation systems used in this study produced apical debris and irrigant extrusion. The reciprocating single-file system tended to produce more debris and irrigant extrusion, compared with the rotary systems. Considering the preparation time, the single-file systems appeared to be advantageous due to their working time.

  7. An investigation on the effects of resection angle on apical sea!

    Directory of Open Access Journals (Sweden)

    Razmi H

    2003-08-01

    Full Text Available Statement of Problem: The goal of a successful endodontic surgery is to establish an adequate seal in apical region. Therefore during surgery, root-end resection is essential for an appropriate situation and proper access."nPurpose: The aim of this study was to determine the most appropriate root-end resection angle to establish adequate apical seal in periapical surgery,"nMaterials and Methods: Forty-five intact extracted human maxillary central incisors were randomly divided into three 15-tooth groups. The root canals were instrumented and then obturated using lateral condensation technique. AM roots were resected at 3mm from apical portion. Angles of root resection for each group were 0, 30 and 45 degrees. Then all teeth surfaces, except the cut ones, were covered with nail polish and immersed in methylen blue for 7 days. After 7 days, nail polish was removed from the surfaces and samples were split in buccolingual direction. Then, dye penetration (mm was measured using a stereozoom microscope (Loupe [X4 magnification] and eye lens. In order to analyze the data, unilateral variance analysis was used."nResults: The values of dye penetration were: 7.23±3.34 (0°, 8.46±3.01(30° and 9.74±4.11 (45°. There"nwas no significant difference between three groups (P>0.05. Conclusion: It is suggested that the angle of root-end resection put a limited effect on apical seal if retrograde cavity depth is increased.

  8. An auxin responsive CLE gene regulates shoot apical meristem development in Arabidopsis

    OpenAIRE

    Guo, Hongyan; Zhang, Wei; Tian, Hainan; Zheng, Kaijie; Dai, Xuemei; Liu, Shanda; Hu, Qingnan; Wang, Xianling; Liu, Bao; Wang, Shucai

    2015-01-01

    Plant hormone auxin regulates most, if not all aspects of plant growth and development, including lateral root formation, organ pattering, apical dominance, and tropisms. Peptide hormones are peptides with hormone activities. Some of the functions of peptide hormones in regulating plant growth and development are similar to that of auxin, however, the relationship between auxin and peptide hormones remains largely unknown. Here we report the identification of OsCLE48, a rice (Oryza sativa) CL...

  9. An auxin responsive CLE gene regulates shoot apical meristem development in Arabidopsis

    OpenAIRE

    Hongyan eGuo; Wei eZhang; Hainan eTian; Kaijie eZheng; Xuemei eDai; Shanda eLiu; Qingnan eHu; Xianling eWang; Bao eLiu; Shucai eWang

    2015-01-01

    Plant hormone auxin regulates most, if not all aspects of plant growth and development, including lateral root formation, organ pattering, apical dominance and tropisms. Peptide hormones are peptides with hormone activities. Some of the functions of peptide hormones in regulating plant growth and development are similar to that of auxin, however, the relationship between auxin and peptide hormones remains largely unknown. Here we report the identification of OsCLE48, a rice (Oryza sativa) CLE...

  10. Sealing ability of grar MTA AngelusTM, CPM TM and MBPc used as apical plugs.

    Science.gov (United States)

    Orosco, Fernando Accorsi; Bramante, Clovis Monteiro; Garcia, Roberto Brandão; Bernadineli, Norberti; Moraes, Ivaldo Gomes de

    2008-01-01

    This study evaluated the sealing ability of apical plugs fabricated with gray MTA Angelus sealer, CPM TM sealer and MBPc sealer. The root canals of 98 extracted single-rooted human teeth were instrumented with #5 to #1 Gates Glidden drills according to the crown-down technique until the #1 drill could pass through the apical foramen. The specimens were then prepared with K-files, starting with an ISO 50 until an ISO 90 could be visualized 1 mm beyond the apex. After root canal preparation, the external surface of each root was rendered impermeable and roots were assigned to 3 experimental groups (n = 30), which received a 5-mm thick apical plug of gray MTA Angelus, CPM and MBPc, and two control groups (n=4). The remaining portion of the canal in the experimental groups was filled by the lateral condensation technique. The teeth of each group, properly identified, were fixed on utility wax by their crowns and were placed in plastic flasks, leaving the apex free and facing upward. The flasks were filled with 0.2% Rhodamine B solution, pH 7.0, so as to completely cover the root apex of all teeth. The sealing ability was analyzed by measuring 0.2% Rhodamine B leakage after all groups had been maintained in this solution for 48 hours. Data were analyzed statistically by Kruskal-Wallis test and Dunn test with a=5%. The results showed that, among the tested materials used for fabrication of apical plugs, MBPc sealer had the least amount of leakage with statistically significant difference (p<0.05). PMID:19089289

  11. Sealing ability of grar MTA AngelusTM, CPM TM and MBPc used as apical plugs

    Directory of Open Access Journals (Sweden)

    Fernando Accorsi Orosco

    2008-02-01

    Full Text Available This study evaluated the sealing ability of apical plugs fabricated with gray MTA AngelusTM sealer, CPM TM sealer and MBPc sealer. The root canals of 98 extracted single-rooted human teeth were instrumented with #5 to #1 Gates Glidden drills according to the crown-down technique until the #1 drill could pass through the apical foramen. The specimens were then prepared with K-files, starting with an ISO 50 until an ISO 90 could be visualized 1 mm beyond the apex. After root canal preparation, the external surface of each root was rendered impermeable and roots were assigned to 3 experimental groups (n = 30, which received a 5-mm thick apical plug of gray MTA AngelusTM, CPM TM and MBPc, and two control groups (n=4. The remaining portion of the canal in the experimental groups was filled by the lateral condensation technique. The teeth of each group, properly identified, were fixed on utility wax by their crowns and were placed in plastic flasks, leaving the apex free and facing upward. The flasks were filled with 0.2% Rhodamine B solution, pH 7.0, so as to completely cover the root apex of all teeth. The sealing ability was analyzed by measuring 0.2% Rhodamine B leakage after all groups had been maintained in this solution for 48 hours. Data were analyzed statistically by Kruskal-Wallis test and Dunn test with a=5%. The results showed that, among the tested materials used for fabrication of apical plugs, MBPc sealer had the least amount of leakage with statistically significant difference (p<0.05.

  12. Expression analysis of Matrix Metalloproteinase-9 in epithelialized and non-epithelialized apical periodontitis lesions

    Science.gov (United States)

    Carneiro, Everdan; Menezes, Renato; Garlet, Gustavo Pompermaier; Garcia, Roberto Brandão; Bramante, Clóvis Monteiro; Figueira, Rita; Sogayar, Mari; Granjeiro, José Mauro

    2009-01-01

    OBJECTIVE To determine the expression of matrix metalloproteinase-9 (MMP-9) in apical periodontitis lesions. STUDY DESIGN Nineteen epithelialized and eighteen non-epithelialized apical periodontitis lesions were collected after periapical surgery. After histological processing, serial sectioning, H&E staining and microscopic analysis, 10 epithelialized and 10 non-epithelialized lesions were selected for immunohistochemical analysis for MMP-9 and CD 68. At least 1/3 of each specimen was frozen at −70°C for further mRNA isolation and reverse transcription into cDNA for Real-Time-PCR procedures. The relative expression of a target gene was determined in comparison with reference genes (GAPDH, HPRT, β-actin and BCRP). RESULTS Polymorphonuclear neutrophils, macrophages and lymphocytes were stained for MMP-9 in both types of lesions, and when present, epithelial cells were also stained. The number and the ratio of MMP-9+/total cells were greater in non-epithelialized than epithelialized lesions (p=0.0001) and showed a positive correlation to CD68+/total cells (p=0.045). No significant differences were observed for MMP-9 mRNA expression between ephithelized and non-ephithelized lesions. However, when compared to healthy periapical ligaments, both types of lesions presented increased MMP-9 expression (p<0.0001). CONCLUSION The present data suggest the participation of several inflammatory cells, mainlly CD68+ cells, in the MMP-9 expression in apical periodontitis lesions. MMP-9 could be actively enroled in the ECM degradation in apical periodontitis lesions. PMID:18926740

  13. DETECTION OF A PUTATIVE 30-KDA LIGAND OF THE CLUSTER-2 ANTIGEN

    NARCIS (Netherlands)

    HELFRICH, W; VANGEEL, M; THE, TH; DELEIJ, L

    1994-01-01

    The cluster-2 antigen, also called EGP-2, is a 38-kDa trans-membrane glycoprotein with a distribution that is largely confined to human epithelial cells and their derived carcinomas. Monoclonal antibodies (MAbs) directed against EGP-2 have been extensively studies as anti-tumors agents, yet the func

  14. Diagnosis and planning in apical surgery: use of cone-beam tomography

    Directory of Open Access Journals (Sweden)

    Regina Karla de Pontes Lima

    2010-10-01

    Full Text Available Introduction and objective: The ability to tridimensionally evaluate pathological and anatomical areas, in apical surgery planning, presents a number of advantages. Cone beam computed tomography (CBCT was developed for dental applications. This paper aims to present a literature review on CBCT, highlighting its advantages over both conventional computed tomography (CT and radiography. Moreover, its clinical applications in apical surgery are discussed. Literature review and conclusion: Unlikely CT, CBCT captures a volume of data in a single 360° rotation, providing benefits such as higher accuracy, better resolution, reduced scanning time and reduced radiation dose. In the maxillofacial region, CBCT has been mainly used in the assessment of dento-alveolar pathology and oral traumatology. CBCT provides a better diagnosis and quantitative information on periodontal bone levels than conventional radiography. It has also been used for patients requiring surgical facial reconstruction, orthognathic surgery, dental implants, and more complex tooth extractions. Besides that, it seems to be a significant tool in modern endodontic practice, presenting useful applications in apical surgery.

  15. Apical Extrusion of Debris Produced during Continuous Rotating and Reciprocating Motion

    Science.gov (United States)

    Nevares, Giselle; Xavier, Felipe; Gominho, Luciana; Cavalcanti, Flávia; Cassimiro, Marcely; Romeiro, Kaline; Alvares, Pamella; Queiroz, Gabriela; Sobral, Ana Paula; Gerbi, Marleny; Silveira, Marcia; Albuquerque, Diana

    2015-01-01

    This study aimed to analyse and compare apical extrusion of debris in canals instrumented with systems used in reciprocating and continuous motion. Sixty mandibular premolars were randomly divided into 3 groups (n = 20): the Reciproc (REC), WaveOne (WO), and HyFlex CM (HYF) groups. One Eppendorf tube per tooth was weighed in advance on an analytical balance. The root canals were instrumented according to the manufacturer's instructions, and standardised irrigation with 2.5% sodium hypochlorite was performed to a total volume of 9 mL. After instrumentation, the teeth were removed from the Eppendorf tubes and incubated at 37°C for 15 days to evaporate the liquid. The tubes were weighed again, and the difference between the initial and final weight was calculated to determine the weight of the debris. The data were statistically analysed using the Shapiro-Wilk, Wilcoxon, and Mann-Whitney tests (α = 5%). All systems resulted in the apical extrusion of debris. Reciproc produced significantly more debris than WaveOne (p < 0.05), and both systems produced a greater apical extrusion of debris than HyFlex CM (p < 0.001). Cross section and motion influenced the results, despite tip standardization. PMID:26543896

  16. Master apical file size - smaller or larger: a systematic review of microbial reduction.

    Science.gov (United States)

    Aminoshariae, A; Kulild, J

    2015-11-01

    The purpose of this systematic review was to determine, in patients undergoing root canal treatment, whether apical enlargement of canals affected microbial reduction. A PICO (population, intervention, comparison and outcome) strategy was developed to identify previously published studies dealing with apical size of canal and microbial reduction. The MEDLINE, Embase, Cochrane and PubMed databases were searched. Additionally, the bibliographies of all relevant articles and textbooks were manually searched. Based on inclusion and exclusion criteria, two reviewers independently selected the relevant articles. Due to the variety of methodologies and different techniques used to measure outcome for master apical file enlargement, it was not possible to standardize the research data and to apply a meta-analysis. Seven articles were identified that met the inclusion criteria. Five of the seven articles generally concluded that canal enlargement reduced bioburden in the root canal system. Two articles reported no difference in canals enlarged to size 25 or 40. The results of the systematic review confirmed that more evidence-based research in this area is needed. With the limited information currently available, the best current available clinical evidence suggests that contemporary chemomechanical debridement techniques with canal enlargement techniques do not eliminate bacteria during root canal treatment at any size.

  17. Sample preparation for laser-microdissection of soybean shoot apical meristem

    Directory of Open Access Journals (Sweden)

    Chui E. Wong

    2012-10-01

    Full Text Available The shoot apical meristem houses stem cells responsible for the continuous formation of aerial plant organs including leaves and stems throughout the life of plants. Laser-microdissection in combination with high-throughput technology such as next generation sequencing permits an in-depth analysis of molecular events associated with specific cell type of interest. Sample preparation is the most critical step in ensuring good quality RNA to be extracted from samples following laser-microdissection. Here, we optimized the sample preparation for a major legume crop, soybean. We used Farmer’s solution as a fixative and paraffin as the embedding medium for soybean shoot apical meristem tissue without the use of any specialized equipment. Shorter time for tissue fixation (two days was found to be critical for the preservation of RNA in soybean shoot apical meristem. We further demonstrated the utility of this method for different tissues derived from soybean and rice. The method outlined here shall facilitate studies on crop plants involving laser-microdissection.

  18. Dynein regulates epithelial polarity and the apical localization of stardust A mRNA.

    Science.gov (United States)

    Horne-Badovinac, Sally; Bilder, David

    2008-01-01

    Intense investigation has identified an elaborate protein network controlling epithelial polarity. Although precise subcellular targeting of apical and basolateral determinants is required for epithelial architecture, little is known about how the individual determinant proteins become localized within the cell. Through a genetic screen for epithelial defects in the Drosophila follicle cells, we have found that the cytoplasmic Dynein motor is an essential regulator of apico-basal polarity. Our data suggest that Dynein acts through the cytoplasmic scaffolding protein Stardust (Sdt) to localize the transmembrane protein Crumbs, in part through the apical targeting of specific sdt mRNA isoforms. We have mapped the sdt mRNA localization signal to an alternatively spliced coding exon. Intriguingly, the presence or absence of this exon corresponds to a developmental switch in sdt mRNA localization in which apical transcripts are only found during early stages of epithelial development, while unlocalized transcripts predominate in mature epithelia. This work represents the first demonstration that Dynein is required for epithelial polarity and suggests that mRNA localization may have a functional role in the regulation of apico-basal organization. Moreover, we introduce a unique mechanism in which alternative splicing of a coding exon is used to control mRNA localization during development.

  19. Dynein regulates epithelial polarity and the apical localization of stardust A mRNA.

    Directory of Open Access Journals (Sweden)

    Sally Horne-Badovinac

    2008-01-01

    Full Text Available Intense investigation has identified an elaborate protein network controlling epithelial polarity. Although precise subcellular targeting of apical and basolateral determinants is required for epithelial architecture, little is known about how the individual determinant proteins become localized within the cell. Through a genetic screen for epithelial defects in the Drosophila follicle cells, we have found that the cytoplasmic Dynein motor is an essential regulator of apico-basal polarity. Our data suggest that Dynein acts through the cytoplasmic scaffolding protein Stardust (Sdt to localize the transmembrane protein Crumbs, in part through the apical targeting of specific sdt mRNA isoforms. We have mapped the sdt mRNA localization signal to an alternatively spliced coding exon. Intriguingly, the presence or absence of this exon corresponds to a developmental switch in sdt mRNA localization in which apical transcripts are only found during early stages of epithelial development, while unlocalized transcripts predominate in mature epithelia. This work represents the first demonstration that Dynein is required for epithelial polarity and suggests that mRNA localization may have a functional role in the regulation of apico-basal organization. Moreover, we introduce a unique mechanism in which alternative splicing of a coding exon is used to control mRNA localization during development.

  20. Effect of nickel titanium file design on the root surface strain and apical microcracks.

    Science.gov (United States)

    Jamleh, Ahmed; Adorno, Carlos G; Ebihara, Arata; Suda, Hideaki

    2016-04-01

    The aim of this study was to determine the effect of nickel titanium file design on the root surface strain generated and apical microcracks caused during canal shaping. Thirty-three mandibular incisors were distributed into LightSpeed X, FlexMaster and a control group. A strain gauge was fixed apically on the proximal root surface to determine the maximum strain during canal shaping. Except for the control group, all root canals were enlarged to size 50. Images were taken after removing the apical 1 and 2 mm of the root end. Mean maximum strain values and presence of microcracks were statistically compared using the t-test and chi-square test, respectively. During canal shaping, the strain increased cumulatively with mean maximum strains of 808.2 ± 228.8 and 525.1 ± 168.9 microstrain in LightSpeed X and FlexMaster, respectively (P = 0.004). Both systems caused comparable microcracks. Although LightSpeed X produced higher maximum strain, no difference in microcrack development was found between both systems.

  1. Effect of nickel titanium file design on the root surface strain and apical microcracks.

    Science.gov (United States)

    Jamleh, Ahmed; Adorno, Carlos G; Ebihara, Arata; Suda, Hideaki

    2016-04-01

    The aim of this study was to determine the effect of nickel titanium file design on the root surface strain generated and apical microcracks caused during canal shaping. Thirty-three mandibular incisors were distributed into LightSpeed X, FlexMaster and a control group. A strain gauge was fixed apically on the proximal root surface to determine the maximum strain during canal shaping. Except for the control group, all root canals were enlarged to size 50. Images were taken after removing the apical 1 and 2 mm of the root end. Mean maximum strain values and presence of microcracks were statistically compared using the t-test and chi-square test, respectively. During canal shaping, the strain increased cumulatively with mean maximum strains of 808.2 ± 228.8 and 525.1 ± 168.9 microstrain in LightSpeed X and FlexMaster, respectively (P = 0.004). Both systems caused comparable microcracks. Although LightSpeed X produced higher maximum strain, no difference in microcrack development was found between both systems. PMID:26420627

  2. Relationship of apical lymph node involvement to survival in resected colon carcinoma.

    Science.gov (United States)

    Malassagne, B; Valleur, P; Serra, J; Sarnacki, S; Galian, A; Hoang, C; Hautefeuille, P

    1993-07-01

    In a prospective study of 197 patients with resected colon carcinoma treated between 1974 and 1985, we explored the relationships between pathologic parameters, and the effect of the latter on survival, to identify the parameter whose systematic measurement would improve the predictive capacity of pathologic staging. Prognostic characteristics were studied by univariate analysis. The results showed significant relationships between the location and number of lymph nodes involved, blood vessel invasion, depth of tumor penetration, and metastases. The five-year survival rates were 45 percent and 17 percent (P < 0.001) for patients without and with apical lymph node involvement, respectively, and 44 percent and 6 percent (P < 0.05) for those with four or less nodes involved and more than four involved, respectively. Among the patients treated by incomplete resection, the respective survival rates of those resected for metastases and of those resected for apical lymph node involvement did not differ significantly. We conclude that the involvement of apical lymph nodes has a significant effect on prognosis and suggest systematic pathologic examination of these nodes to allow simpler and more reproducible selection of patients for treatment by incomplete resection who are at high risk of disease-related death. PMID:8394236

  3. Hypertrophic cardiomyopathy with midventricular obstruction and apical aneurysm formation in a single family: case report

    Directory of Open Access Journals (Sweden)

    Paraskevaidis Stylianos

    2009-06-01

    Full Text Available Abstract Background Hypertrophic cardiomyopathy (HCM is an extremely heterogeneous disease. An under recognized and very often missed subgroup within this broad spectrum concerns patients with left ventricular (LV apical aneurysms in the absence of coronary artery disease. Case presentation We describe a case of HCM with midventricular obstruction and apical aneurysm formation in 3 patients coming from a single family. This HCM pattern was detected by 2D-echocardiography and confirmed by cardiac magnetic resonance imaging. A cardioverter defibrillator was implanted in one of the patients because of non-sustained ventricular tachycardia detected in 24-h Holter monitoring and an abrupt drop in systolic blood pressure during maximal exercise test. The defibrillator activated 8 months after implantation by suppression of a ventricular tachycardia providing anti-tachycardia pacing. The patient died due to refractory heart failure 2 years after initial evaluation. The rest of the patients are stable after a 2.5-y follow-up period. Conclusion The detection of apical aneurysm by echocardiography in HCM patients may be complicated. Ventricular tachycardia arising from the scarred aneurysm wall may often occur predisposing to sudden death.

  4. Integrins Regulate Apical Constriction via Microtubule Stabilization in the Drosophila Eye Disc Epithelium

    Directory of Open Access Journals (Sweden)

    Vilaiwan M. Fernandes

    2014-12-01

    Full Text Available During morphogenesis, extracellular signals trigger actomyosin contractility in subpopulations of cells to coordinate changes in cell shape. To illuminate the link between signaling-mediated tissue patterning and cytoskeletal remodeling, we study the progression of the morphogenetic furrow (MF, the wave of apical constriction that traverses the Drosophila eye imaginal disc preceding photoreceptor neurogenesis. Apical constriction depends on actomyosin contractility downstream of the Hedgehog (Hh and bone morphogenetic protein (BMP pathways. We identify a role for integrin adhesion receptors in MF progression. We show that Hh and BMP regulate integrin expression, the loss of which disrupts apical constriction and slows furrow progression; conversely, elevated integrins accelerate furrow progression. We present evidence that integrins regulate MF progression by promoting microtubule stabilization, since reducing microtubule stability rescues integrin-mediated furrow acceleration. Thus, integrins act as a genetic link between tissue-level signaling events and morphological change at the cellular level, leading to morphogenesis and neurogenesis in the eye.

  5. In vitro analysis of the apical deformation, using a different number of files during the instrumentation

    Directory of Open Access Journals (Sweden)

    Lucas da Fonseca Roberti GARCIA

    2009-12-01

    Full Text Available Introduction and objective: The aim of the present study was toevaluate in vitro the apical deformation in roots of crooked lowerincisors with mesio-distal flattening, using a different number of files during the instrumentation. Material and methods: Twenty rootswere randomly divided into two groups (n = 10. Group I: roots wereinitially instrumented with a K #15 file until a #35 file, through stepback technique. During instrumentation, the canals were irrigated with 1 mL of 1% NaOCl between every instrument change. Obturation wasperformed with Grossman sealer by lateral condensation technique.Group II: procedure was similar to group I, except for the files number, using file #40 as memory instrument. After that, each root was 1mm apical weared in order to reveal the filling. A 3 mm cut from the root apex was performed, resulting in samples with the same length, which were examined with an optical microscope (X40. The images captured by the Image Tool software were analyzed and the area occupied by sealer, gutta-percha and possible empty spaces was quantified. Results: Data was submitted to statistical analysis and showed no statistical significant differences between groups I and II (p > 0.05. Conclusion: The variation in the number of files during instrumentation did not promote apical deformation, but resulted in a similar hermetic sealing between the studied groups.

  6. Thalamic input to distal apical dendrites in neocortical layer 1 is massive and highly convergent.

    Science.gov (United States)

    Rubio-Garrido, Pablo; Pérez-de-Manzo, Flor; Porrero, César; Galazo, Maria J; Clascá, Francisco

    2009-10-01

    Input to apical dendritic tufts is now deemed crucial for associative learning, attention, and similar "feedback" interactions in the cerebral cortex. Excitatory input to apical tufts in neocortical layer 1 has been traditionally assumed to be predominantly cortical, as thalamic pathways directed to this layer were regarded relatively scant and diffuse. However, the sensitive tracing methods used in the present study show that, throughout the rat neocortex, large numbers (mean approximately 4500/mm(2)) of thalamocortical neurons converge in layer 1 and that this convergence gives rise to a very high local density of thalamic terminals. Moreover, we show that the layer 1-projecting neurons are present in large numbers in most, but not all, motor, association, limbic, and sensory nuclei of the rodent thalamus. Some layer 1-projecting axons branch to innervate large swaths of the cerebral hemisphere, whereas others arborize within only a single cortical area. Present data imply that realistic modeling of cortical circuitry should factor in a dense axonal canopy carrying highly convergent thalamocortical input to pyramidal cell apical tufts. In addition, they are consistent with the notion that layer 1-projecting axons may be a robust anatomical substrate for extensive "feedback" interactions between cortical areas via the thalamus.

  7. Supraventricular tachycardia in a patient with Lown-Ganong-Levine syndrome associated with apical hypertrophic cardiomyopathy.

    Science.gov (United States)

    Hayano, M; Imamura, Y; Tsuruta, M; Inoue, J; Nakashima, H; Fukuyama, K; Eguchi, Y; Tsuji, S; Matsuo, S; Yano, K

    1988-03-01

    Electrophysiologic study of a 55-year-old patient with Lown-Ganong-Levine syndrome associated with apical hypertrophic cardiomyopathy is reported. The patient had a history of recurrent attacks of tachyarrhythmia and his electrocardiogram showed a short P-R interval (0.10 sec) with narrow QRS complex and left ventricular hypertrophy with giant negative T waves. His cineangiogram showed severe apical hypertrophy. An electrophysiologic study was performed. The results of programmed atrial pacing show the existence of the dual A-V nodal pathways. The A-H interval at rapid atrial pacing increased maximally by 103 msec. Atrial stimulation could depolarize parts of the atrium without altering the supraventricular tachycardia. These findings suggested that preferential rapidly conducting A-V nodal and intranodal reentry are the responsible mechanisms in this reciprocating tachycardia. We conclude that the short P-R interval was due to intranodal reentry through the dual A-V nodal pathways. To our knowledge, a case of Lown-Ganong-Levine syndrome with apical hypertrophic cardiomyopathy has not been previously described in the literature.

  8. Retromer controls epithelial cell polarity by trafficking the apical determinant Crumbs.

    Science.gov (United States)

    Pocha, Shirin Meher; Wassmer, Thomas; Niehage, Christian; Hoflack, Bernard; Knust, Elisabeth

    2011-07-12

    The evolutionarily conserved apical determinant Crumbs (Crb) is essential for maintaining apicobasal polarity and integrity of many epithelial tissues [1]. Crb levels are crucial for cell polarity and homeostasis, yet strikingly little is known about its trafficking or the mechanism of its apical localization. Using a newly established, liposome-based system described here, we determined Crb to be an interaction partner and cargo of the retromer complex. Retromer is essential for the retrograde transport of numerous transmembrane proteins from endosomes to the trans-Golgi network (TGN) and is conserved between plants, fungi, and animals [2]. We show that loss of retromer function results in a substantial reduction of Crb in Drosophila larvae, wing discs, and the follicle epithelium. Moreover, loss of retromer phenocopies loss of crb by preventing apical localization of key polarity molecules, such as atypical protein kinase C (aPKC) and Par6 in the follicular epithelium, an effect that can be rescued by overexpression of Crb. Additionally, loss of retromer results in multilayering of the follicular epithelium, indicating that epithelial integrity is severely compromised. Our data reveal a mechanism for Crb trafficking by retromer that is vital for maintaining Crb levels and localization. We also show a novel function for retromer in maintaining epithelial cell polarity. PMID:21700461

  9. Influence of Instruments Used in Root Canal Preparation on Amount of Apically Extruded Debris.

    Science.gov (United States)

    Karataş, Ertuğrul; Ersoy, İbrahim; Gündüz, Hicran Ateş; Uygun, Ahmet Demirhan; Kol, Elif; Çakıcı, Fatih

    2016-08-01

    The aim of the present study was to compare the effects of ProTaper Gold, WaveOne Gold, ProTaper Universal, and WaveOne instruments on the amount of apically extruded debris. Eighty mandibular premolar teeth with straight root canals were selected and assigned to four groups (n = 20). The root canals were instrumented using ProTaper Gold, WaveOne Gold, ProTaper Universal, and WaveOne systems. Eppendorf tubes containing apically extruded debris were weighed three times, and mean values were calculated. The net mass of the extruded debris was calculated by subtracting the initial mass from the final mass. The groups were compared using one-way analysis of variance and Tukey's post hoc tests at a significance level of P < 0.05. The PTG group extruded less debris than the PTU group, and the WOG group extruded less debris than the WO group (P < 0.05). All the instrumentation systems tested in the present study were associated with apical extrusion of debris. PMID:26814380

  10. Takotsubo, discinesia apical transitoria: Presentacion de 4 casos y revisión de la literatura Takotsubo, reversible apical ballooning of the left ventricle: Report of 4 cases

    Directory of Open Access Journals (Sweden)

    Bárbara C. Finn

    2005-10-01

    Full Text Available En 1990 se describió un síndrome caracterizado por discinesia apical transitoria, cambios en el electrocardiograma (ECG, mínima elevación de enzimas cardíacas y arterias coronarias normales. Se presenta con mayor frecuencia en mujeres añosas y es precedido por un evento de estrés, ya sea físico o psíquico. La fisiopatología aún no está aclarada y se proponen diversas teorías. La de mayor peso es la que postula una afección secundaria a la descarga de catecolaminas desencadenada por el estrés, sobre un corazón incapaz de mantener una respuesta inotrópica adecuada. Se presentan cuatro casos de pacientes atendidos en nuestro hospital que se manifestaron con síntomas sugerentes de infarto agudo de miocardio asociados a insuficiencia cardiaca, en el contexto de un episodio estresante. Los síntomas preponderantes fueron dolor precordial opresivo y disnea. En el ECG se evidenciaron tanto supradesnivel del segmento ST, como inversión de la onda T. Todos los pacientes presentaron la imagen ecocardiográfica típica de discinesia apical, y todos tuvieron en la cinecoronariografía coronarias normales. Cabe destacar que tanto el ECG, como el ecocardiograma volvieron a la normalidad a partir de las dos semanas.A syndrome of apical ballooning, with ECG mimicking acute myocardial infarctation, mild or no enzymatic changes, and normal coronary angiogram was described in 1990. It presents mainly in middle aged and elderly women and it is preceded by stress triggering circumstances. Several mechanisms have been proposed although the precise cause remains unclear. The most accepted theory proposes the interaction of catecholamines and an inadequate inotropic response. We report four cases that presented with chest pain and sudden onset of heart failure, all patients had physical or emotional stress as a triggering factor. On ECG, ST segment elevation and inverted T waves were observed in the acute phase. All patients had typical echocardiogram

  11. [Deformations occurring in the apical third of curved root canals during biomechanical preparation using manual impulsion-traction techniques].

    Science.gov (United States)

    Roig Cayón, M; Basilio Monné, J; Canalda Sahli, C

    1990-01-01

    Apical deformations, specially zips and elbows, during instrumentation of the root canals, are studied. The authors study why do they appear, their effect on endodontic therapy, and the way of avoiding them. PMID:1964069

  12. [Deformations occurring in the apical third of curved root canals during biomechanical preparation using manual impulsion-traction techniques].

    Science.gov (United States)

    Roig Cayón, M; Basilio Monné, J; Canalda Sahli, C

    1990-01-01

    Apical deformations, specially zips and elbows, during instrumentation of the root canals, are studied. The authors study why do they appear, their effect on endodontic therapy, and the way of avoiding them.

  13. Epstein-Barr virus infection induces bone resorption in apical periodontitis via increased production of reactive oxygen species.

    Science.gov (United States)

    Jakovljevic, Aleksandar; Andric, Miroslav; Miletic, Maja; Beljic-Ivanovic, Katarina; Knezevic, Aleksandra; Mojsilovic, Slavko; Milasin, Jelena

    2016-09-01

    Chronic inflammatory processes in periapical tissues caused by etiological agents of endodontic origin lead to apical periodontitis. Apart from bacteria, two herpesviruses, Epstein-Barr virus (EBV) and Human cytomegalovirus (HCMV) are recognized as putative pathogens in apical periodontitis. Although previous reports suggest the involvement of EBV in the pathogenesis of apical periodontitis, its exact role in periapical bone resorption has not yet been fully elucidated. We hypothesize that EBV infection in apical periodontitis is capable of inducing periapical bone resorption via stimulation of reactive oxygen species (ROS) overproduction. Increased levels of ROS induce expression of receptor activator of nuclear factor kappa B (NF-κB) ligand (RANKL). RANKL binding to receptor activator of nuclear factor κB (RANK) present on the surface of preosteoclasts induces their maturation and activation which consequently leads to bone resorption. The potential benefit of antiviral and antioxidant-based therapies in periapical bone resorption treatment remains to be assessed. PMID:27515196

  14. Natural selection promotes antigenic evolvability

    NARCIS (Netherlands)

    Graves, C.J.; Ros, V.I.D.; Stevenson, B.; Sniegowski, P.D.; Brisson, D.

    2013-01-01

    The hypothesis that evolvability - the capacity to evolve by natural selection - is itself the object of natural selection is highly intriguing but remains controversial due in large part to a paucity of direct experimental evidence. The antigenic variation mechanisms of microbial pathogens provide

  15. Oncogenic cancer/testis antigens

    DEFF Research Database (Denmark)

    Gjerstorff, Morten F; Andersen, Mads H; Ditzel, Henrik J

    2015-01-01

    /testis antigens are immunogenic, highly cancer-specific, and frequently expressed in various types of cancer, which make them promising candidate targets for cancer immunotherapy, including cancer vaccination and adoptive T-cell transfer with chimeric T-cell receptors. Our current understanding of tumor...

  16. The effect of bevel angle on apical microleakage following the use of amalgam and MTA

    Directory of Open Access Journals (Sweden)

    Sharifian MR

    2006-07-01

    Full Text Available Background and Aim: Achieving appropriate seal in canal obturation is the main goal of endodontic therapy. However, in some cases, it can not be obtained by non-surgical procedures alone. Retrograde surgery is one of the most common procedures in endodontics. Apical seal improvement can be obtained by root end filling and decreasing the root end resection angle (bevel angle. The aim of this study was to investigate the role of bevel angle on apical microleakage following the use of amalgam and MTA as root end filling materials. Materials and Methods: In this experimental study, 128 extracted human teeth were instrumented and obturated by lateral condensation technique. The teeth were divided into two groups and the apical root resection was performed by high speed fissure bur (one group perpendicular to the long axis of the teeth and the other 45 to the long axis. The 3 mm root end cavity was prepared by ultrasonic device. Each group was divided into two subgroups: One filled with amalgam and the other with MTA. Teeth were incubated for 72 h, covered by two layers of nail polish (except for apical 3mm and submerged in methylene blue for 48 h. Teeth were washed under tap water and mesiodistally dissected by low speed disc. Dye penetration was evaluated by stereomicroscope. Data were analysed by two way ANOVA and Tukey tests with p<0.05 as the limit of significance. Results: The results showed that retrofill material type had a significant effect on microleakage and MTA was superior to amalgam in this respect. Bevel angle failed to show any significant effect on apical microleakage. Conclusion: Based on the results of this study, it seems that the use of MTA instead of amalgam in clinical practice can improve the success rate of endodontic surgery whereas the bevel angle can be determined based on the status of each individual case; However, increasing the bevel angle seems to increase microleakage due to exposure of more dentinal tubules.

  17. Human leukocyte antigen (HLA)-G during pregnancy part I

    DEFF Research Database (Denmark)

    Klitkou, Louise; Dahl, Mette; Hviid, Thomas Vauvert F;

    2015-01-01

    Human leukocyte antigen (HLA)-G is a class Ib molecule with restricted tissue distribution expressed on trophoblast cells and has been proposed to have immunomodulatory functions during pregnancy. Soluble HLA-G1 (sHLA-G1) can be generated by the shedding of membrane-bound HLA-G molecules; however...... of importance for production of sHLA-G in the mother and child, or it may support the theory that sHLA-G in the pregnant woman and the fetus is partly derived from a "shared organ", the placenta....

  18. Apical ballooning syndrome complicated by acute severe mitral regurgitation with left ventricular outflow obstruction – Case report

    OpenAIRE

    Celermajer David S; Chandrasegaram Manju D; Wilson Michael K

    2007-01-01

    Abstract Background Apical ballooning syndrome (or Takotsubo cardiomyopathy) is a syndrome of transient left ventricular apical ballooning. Although first described in Japanese patients, it is now well reported in the Caucasian population. The syndrome mimicks an acute myocardial infarction but is characterised by the absence of obstructive coronary disease. We describe a serious and poorly understood complication of Takotsubo cardiomyopathy. Case Presentation We present the case of a 65 year...

  19. Robotic membranes

    DEFF Research Database (Denmark)

    Ramsgaard Thomsen, Mette

    2008-01-01

    , Vivisection and Strange Metabolisms, were developed at the Centre for Information Technology and Architecture (CITA) at the Royal Danish Academy of Fine Arts in Copenhagen as a means of engaging intangible digital data with tactile physical material. As robotic membranes, they are a dual examination...

  20. Biological function and molecular mapping of M antigen in yeast phase of Histoplasma capsulatum.

    Directory of Open Access Journals (Sweden)

    Allan Jefferson Guimarães

    Full Text Available Histoplasmosis, due to the intracellular fungus Histoplasma capsulatum, can be diagnosed by demonstrating the presence of antibodies specific to the immunodominant M antigen. However, the role of this protein in the pathogenesis of histoplasmosis has not been elucidated. We sought to structurally and immunologically characterize the protein, determine yeast cell surface expression, and confirm catalase activity. A 3D-rendering of the M antigen by homology modeling revealed that the structures and domains closely resemble characterized fungal catalases. We generated monoclonal antibodies (mAbs to the protein and determined that the M antigen is present on the yeast cell surface and in cell wall/cell membrane preparations. Similarly, we found that the majority of catalase activity was in extracts containing fungal surface antigens and that the M antigen is not significantly secreted by live yeast cells. The mAbs also identified unique epitopes on the M antigen. The localization of the M antigen to the cell surface of H. capsulatum yeast and the characterization of the protein's major epitopes have important implications since it demonstrates that although the protein may participate in protecting the fungus against oxidative stress it is also accessible to host immune cells and antibody.

  1. Concepts and applications for influenza antigenic cartography

    Science.gov (United States)

    Cai, Zhipeng; Zhang, Tong; Wan, Xiu-Feng

    2011-01-01

    Influenza antigenic cartography projects influenza antigens into a two or three dimensional map based on immunological datasets, such as hemagglutination inhibition and microneutralization assays. A robust antigenic cartography can facilitate influenza vaccine strain selection since the antigenic map can simplify data interpretation through intuitive antigenic map. However, antigenic cartography construction is not trivial due to the challenging features embedded in the immunological data, such as data incompleteness, high noises, and low reactors. To overcome these challenges, we developed a computational method, temporal Matrix Completion-Multidimensional Scaling (MC-MDS), by adapting the low rank MC concept from the movie recommendation system in Netflix and the MDS method from geographic cartography construction. The application on H3N2 and 2009 pandemic H1N1 influenza A viruses demonstrates that temporal MC-MDS is effective and efficient in constructing influenza antigenic cartography. The web sever is available at http://sysbio.cvm.msstate.edu/AntigenMap. PMID:21761589

  2. Mineral trioxide aggregate apical plug method for the treatment of nonvital immature permanent maxillary incisors: Three case reports

    Directory of Open Access Journals (Sweden)

    Betül Günes

    2012-01-01

    Full Text Available Treatment of nonvital immature permanent teeth with calcium-hydroxide is associated with some difficulties such as weakened tooth fracture, root canal reinfection and long treatment time. Mineral trioxide aggregate (MTA apical plug method is an alternative treatment option for open apices, and has gained popularity in the recent times. In this case report, we have attempted to present successful treatment of three maxillary incisors with open apices and periapical lesions with MTA. After preparing the access cavity, the working length was determined. The root canals were irrigated with 2.5% Sodium hypochlorite (NaOCl and disinfected with calcium-hydroxide for two weeks. MTA was then placed in the apical 3 millimeters of the root canal. The remaining part of the root canal was filled with gutta-percha and the coronal restoration was finished with composite resin. After six months the radiographic examination showed a decrease of periapical lesions. At a 1-year and 18-months follow up, radiological and clinical successful healing of the incisor teeth was seen. MTA seems as an effective material for the apical plug method for the treatment of nonvital immature permanent teeth with open apices.

  3. Fractal dimension of apical dendritic arborization differs in the superficial and the deep pyramidal neurons of the rat cerebral neocortex.

    Science.gov (United States)

    Puškaš, Nela; Zaletel, Ivan; Stefanović, Bratislav D; Ristanović, Dušan

    2015-03-01

    Pyramidal neurons of the mammalian cerebral cortex have specific structure and pattern of organization that involves the presence of apical dendrite. Morphology of the apical dendrite is well-known, but quantification of its complexity still remains open. Fractal analysis has proved to be a valuable method for analyzing the complexity of dendrite morphology. The aim of this study was to establish the fractal dimension of apical dendrite arborization of pyramidal neurons in distinct neocortical laminae by using the modified box-counting method. A total of thirty, Golgi impregnated neurons from the rat brain were analyzed: 15 superficial (cell bodies located within lamina II-III), and 15 deep pyramidal neurons (cell bodies situated within lamina V-VI). Analysis of topological parameters of apical dendrite arborization showed no statistical differences except in total dendritic length (p=0.02), indicating considerable homogeneity between the two groups of neurons. On the other hand, average fractal dimension of apical dendrite was 1.33±0.06 for the superficial and 1.24±0.04 for the deep cortical neurons, showing statistically significant difference between these two groups (pfractal dimension values, apical dendrites of the superficial pyramidal neurons tend to show higher structural complexity compared to the deep ones.

  4. Sudden Cardiac Arrest in a Patient With Apical Hypertrophic Cardiomyopathy: Case Report and a Brief Review of Literature.

    Science.gov (United States)

    Gupta, Tanush; Paul, Neha; Palaniswamy, Chandrasekar; Balasubramaniyam, Nivas; Aronow, Wilbert S; Kolte, Dhaval; Khera, Sahil; Shah, Amar B; Gass, Alan

    2016-01-01

    Apical hypertrophic cardiomyopathy (HCM) is a phenotypic variant of nonobstructive HCM, in which hypertrophy of the myocardium predominantly involves the left ventricular apex. It is common in Japanese and other Asian populations but is rare in the United States. Apical HCM has a relatively benign prognosis in terms of cardiovascular mortality; however, morbid events such as ventricular aneurysms, apical thrombi, diastolic dysfunction, atrial fibrillation, and myocardial infarction are not uncommon. We report a case of an 18-year-old white man who presented to our hospital after an out-of-hospital cardiac arrest. The patient had a witnessed collapse while playing basketball in the field. He was found to be pulseless and unresponsive by his coach, and cardiopulmonary resuscitation was immediately started. Upon arrival of emergency medical services, an automated external defibrillator advised shock and he was defibrillated thrice. Return of spontaneous circulation was achieved in 15 minutes. He was intubated for airway protection and was brought to the hospital. Therapeutic hypothermia was initiated. He demonstrated good neurological status after active rewarming. Subsequent cardiac magnetic resonance imaging was suggestive of apical HCM with right ventricular involvement. The patient underwent an implantable cardioverter defibrillator placement for secondary prevention and was subsequently discharged. In conclusion, apical HCM can rarely be associated with adverse cardiovascular events. The diagnosis may be missed on transthoracic 2-dimensional cardiac echocardiogram, and cardiac magnetic resonance imaging should be considered to exclude apical HCM in young patients who present after sudden cardiac arrest. PMID:25923227

  5. Genome Scale Identification of Treponema pallidum Antigens

    OpenAIRE

    McKevitt, Matthew; Brinkman, Mary Beth; McLoughlin, Melanie; Perez, Carla; Howell, Jerrilyn K.; Weinstock, George M.; Norris, Steven J; Palzkill, Timothy

    2005-01-01

    Antibody responses for 882 of the 1,039 proteins in the proteome of Treponema pallidum were examined. Sera collected from infected rabbits were used to systematically identify 106 antigenic proteins, including 22 previously identified antigens and 84 novel antigens. Additionally, sera collected from rabbits throughout the course of infection demonstrated a progression in the breadth and intensity of humoral immunoreactivity against a representative panel of T. pallidum antigens.

  6. A method for visualizing surface-exposed and internal PfEMP1 adhesion antigens in Plasmodium falciparum infected erythrocytes

    DEFF Research Database (Denmark)

    Bengtsson, Dominique; Sowa, Kordai M; Salanti, Ali;

    2008-01-01

    BACKGROUND: The insertion of parasite antigens into the host erythrocyte membrane and the structure and distribution of Plasmodium falciparum adhesion receptors on that membrane are poorly understood. Laser scanning confocal microscopy (LSCM) and a novel labelling and fixation method have been used......-ionic detergent to permit access of antibodies to internal parasite antigens. Differentiation between surface and internal antigens is achieved using antibodies labelled with different fluorochromes and confocal microscopy RESULTS: Surface exposed PfEMP1 is first detectable by antibodies at the trophozoite stage...

  7. Rapid and specific biotin labelling of the erythrocyte surface antigens of both cultured and ex-vivo Plasmodium parasites

    Directory of Open Access Journals (Sweden)

    Thompson Joanne

    2007-05-01

    Full Text Available Abstract Background Sensitive detection of parasite surface antigens expressed on erythrocyte membranes is necessary to further analyse the molecular pathology of malaria. This study describes a modified biotin labelling/osmotic lysis method which rapidly produces membrane extracts enriched for labelled surface antigens and also improves the efficiency of antigen recovery compared with traditional detergent extraction and surface radio-iodination. The method can also be used with ex-vivo parasites. Methods After surface labelling with biotin in the presence of the inhibitor furosemide, detergent extraction and osmotic lysis methods of enriching for the membrane fractions were compared to determine the efficiency of purification and recovery. Biotin-labelled proteins were identified on silver-stained SDS-polyacrylamide gels. Results Detergent extraction and osmotic lysis were compared for their capacity to purify biotin-labelled Plasmodium falciparum and Plasmodium chabaudi erythrocyte surface antigens. The pellet fraction formed after osmotic lysis of P. falciparum-infected erythrocytes is notably enriched in suface antigens, including PfEMP1, when compared to detergent extraction. There is also reduced co-extraction of host proteins such as spectrin and Band 3. Conclusion Biotinylation and osmotic lysis provides an improved method to label and purify parasitised erythrocyte surface antigen extracts from both in vitro and ex vivo Plasmodium parasite preparations.

  8. Antigen Export Reduces Antigen Presentation and Limits T Cell Control of M. tuberculosis.

    Science.gov (United States)

    Srivastava, Smita; Grace, Patricia S; Ernst, Joel D

    2016-01-13

    Persistence of Mycobacterium tuberculosis results from bacterial strategies that manipulate host adaptive immune responses. Infected dendritic cells (DCs) transport M. tuberculosis to local lymph nodes but activate CD4 T cells poorly, suggesting bacterial manipulation of antigen presentation. However, M. tuberculosis antigens are also exported from infected DCs and taken up and presented by uninfected DCs, possibly overcoming this blockade of antigen presentation by infected cells. Here we show that the first stage of this antigen transfer, antigen export, benefits M. tuberculosis by diverting bacterial proteins from the antigen presentation pathway. Kinesin-2 is required for antigen export and depletion of this microtubule-based motor increases activation of antigen-specific CD4 T cells by infected cells and improves control of intracellular infection. Thus, although antigen transfer enables presentation by bystander cells, it does not compensate for reduced antigen presentation by infected cells and represents a bacterial strategy for CD4 T cell evasion.

  9. Chitosan cushioned phospholipid membrane and its application in imaging ellipsometry based-biosensor

    International Nuclear Information System (INIS)

    Chitosan cushion can support the air stability of phospholipid membrane, but the problem of serum solubility of phospholipid membrane prevents it from use in serum detection applications. Poly (ethylene glycol) (PEG) shielding promises both stability and non-specific adsorption resistance for phospholipid membrane. An air stable phospholipid membrane microarray has been successfully fabricated on chitosan modified silicon wafer. We have demonstrated the potential application of PEGylated phospholipid membrane in imaging ellipsometry-based protein biosensor. Because of the strong resistance against non-specific adsorption of serum, antigens are immobilized onto the membrane surface through chemical activation and further bind their antibodies without using blocking agent. Taking advantage of the multiple and parallel reaction capabilities of microfluidic reactor system, we have assayed the binding by varying both the density of antigen on the membrane surface and the concentration of antibody in solution.

  10. [HLA antigens in juvenile rheumatoid arthritis].

    Science.gov (United States)

    Rumba, I V; Sochnev, A M; Kukaĭne, E M; Burshteĭn, A M; Benevolenskaia, L I

    1990-01-01

    Antigens of I class HLA system (locus A and B) were investigated in 67 patients of Latvian nationality suffering from juvenile rheumatoid arthritis (JRA). Associations of HLA antigens with juvenile rheumatoid arthritis partially coincided with the ones revealed earlier. Typing established an increased incidence of antigen B27 (p less than 0.01) and gaplotype A2, B40 (p less than 0.01). Antigen B15 possessed a protective action with respect to JRA. Interlocus combinations demonstrated a closer association with the disease than a single antigen. The authors also revealed markers of various clinico-anatomical variants of JRA.

  11. Basal and apical regulation of VEGF-A and placenta growth factor in the RPE/choroid and primary RPE

    Science.gov (United States)

    Kaya, Leyla; Flach, Janina; Lassen, Jens; Treumer, Felix; Roider, Johann

    2015-01-01

    Purpose Members of the vascular endothelial growth factor (VEGF) family are strongly involved in pathological processes in the retina, such as age-related macular degeneration and diabetic retinopathy. Cells of the retinal pigment epithelium (RPE) constitutively secrete VEGF-A, and the secretion of placental growth factor (PlGF) has also been described. RPE cells are strongly polarized cells with different secretome at the apical and basal side. In this study, we evaluated the basal and apical regulation of VEGF-A and PlGF secretion in RPE/choroid explants and primary RPE cells. Methods RPE/choroid tissue explants were prepared from porcine eyes and cultivated in modified Ussing chambers, separating apical (RPE) and basal (choroid) supernatant. Primary RPE cells were also prepared from porcine eyes and cultivated on Transwell plates. Explants and cells were treated with inhibitors for VEGFR-2 (SU1498), p38 (SB203580), and the transcription factors nuclear factor-kappa B (NF-κB) and SP-1 (mithramycin), respectively. VEGF-A and PlGF content was evaluated with enzyme-linked immunosorbent assay (ELISA). In addition, western blots were performed. Results In the RPE/choroid, VEGF-A can initially be found on the apical and basal sides with significantly more pronounced secretion on the basal side. VEGF-A secretion is differentially regulated on the apical and basal sides, with the inhibition of SP-1 and NF-κB showing strong effects apically and basally after 24 h and 48 h, the inhibition of p38 displaying its effect mainly on the basal side with some effect apically after 48 h, and the inhibition of VEGFR-2 reducing the secretion of VEGF only on the apical side at 24 h and 48 h. In the RPE cell culture, similar effects were found, with inhibition of NF-κB or SP-1 displaying a strong decrease in VEGF-A on both sides, and p38 inhibition displaying only an inhibitory effect on the basal side. In contrast, an apical effect of VEGFR-2 inhibition was not found. However, the

  12. Analysis of dental root apical morphology: a new method for dietary reconstructions in primates.

    Science.gov (United States)

    Hamon, NoÉmie; Emonet, Edouard-Georges; Chaimanee, Yaowalak; Guy, Franck; Tafforeau, Paul; Jaeger, Jean-Jacques

    2012-06-01

    The reconstruction of paleo-diets is an important task in the study of fossil primates. Previously, paleo-diet reconstructions were performed using different methods based on extant primate models. In particular, dental microwear or isotopic analyses provided accurate reconstructions for some fossil primates. However, there is sometimes difficult or impossible to apply these methods to fossil material. Therefore, the development of new, independent methods of diet reconstructions is crucial to improve our knowledge of primates paleobiology and paleoecology. This study aims to investigate the correlation between tooth root apical morphology and diet in primates, and its potential for paleo-diet reconstructions. Dental roots are composed of two portions: the eruptive portion with a smooth and regular surface, and the apical penetrative portion which displays an irregular and corrugated surface. Here, the angle formed by these two portions (aPE), and the ratio of penetrative portion over total root length (PPI), are calculated for each mandibular tooth root. A strong correlation between these two variables and the proportion of some food types (fruits, leaves, seeds, animal matter, and vertebrates) in diet is found, allowing the use of tooth root apical morphology as a tool for dietary reconstructions in primates. The method was then applied to the fossil hominoid Khoratpithecus piriyai, from the Late Miocene of Thailand. The paleo-diet deduced from aPE and PPI is dominated by fruits (>50%), associated with animal matter (1-25%). Leaves, vertebrates and most probably seeds were excluded from the diet of Khoratpithecus, which is consistent with previous studies. PMID:22553124

  13. Factors Influencing Selection of Vaginal, Open Abdominal, or Robotic Surgery to Treat Apical Vaginal Vault Prolapse

    Science.gov (United States)

    Anand, Mallika; Weaver, Amy L.; Fruth, Kristin M.; Gebhart, John B.

    2016-01-01

    Objectives To determine factors influencing selection of Mayo-McCall culdoplasty (MMC), open abdominal sacrocolpopexy (ASC), or robotic sacrocolpopexy (RSC) for posthysterectomy vaginal vault prolapse. Methods We retrospectively searched for the records of patients undergoing posthysterectomy apical vaginal prolapse surgery between January 1, 2000, and June 30, 2012, at our institution. Baseline characteristics and explicit selection factors were abstracted from the electronic medical records. Factors were compared between groups using χ2 tests for categorical variables, ANOVA for continuous variables, and Kruskal-Wallis tests for ordinal variables. Results Among the 512 patients identified who met inclusion criteria, the MMC group (n=174) had more patients who were older, American Society of Anesthesiologists class 3+ or greater, had anterior vaginal prolapse grade 3+, desired to avoid abdominal surgery, and did not desire a functional vagina. Patients in the ASC (n=237) and RSC (n=101) groups had more failed prolapse surgeries, suspected abdominopelvic pathologic processes, and chronic pain. Advanced prolapse was more frequently cited as an explicit selection factor for ASC than for either MMC or RSC. Conclusions The most common factors that influenced the type of apical vaginal vault prolapse surgery overlapped with characteristics that differed at baseline. In general, MMC was chosen for advanced anterior vaginal prolapse and baseline characteristics that increased surgical risks, ASC for advanced apical prolapse, and ASC or RSC for recurrent prolapse, suspected abdominal pathology, and patients with chronic pain or lifestyles including heavy lifting. Thus, efforts should be made to attempt to control for selection bias when comparing these procedures. PMID:26945273

  14. Apices of maxillary premolars observed by swept source optical coherence tomography

    Science.gov (United States)

    Ebihara, Arata; Iino, Yoshiko; Yoshioka, Toshihiko; Hanada, Takahiro; Sunakawa, Mitsuhiro; Sumi, Yasunori; Suda, Hideaki

    2015-02-01

    Apicoectomy is performed for the management of apical periodontitis when orthograde root canal treatment is not possible or is ineffective. Prior to the surgery, cone beam computed tomography (CBCT) examination is often performed to evaluate the lesion and the adjacent tissues. During the surgical procedure, the root apex is resected and the resected surface is usually observed under dental operating microscope (DOM). However, it is difficult to evaluate the details and the subsurface structure of the root using CBCT and DOM. A new diagnostic system, swept source optical coherence tomography (SS-OCT), has been developed to observe the subsurface anatomical structure. The aim of this study was to observe resected apical root canals of human maxillary premolars using SS-OCT and compare the findings with those observed using CBCT and DOM. Six extracted human maxillary premolars were used. After microfocus computed tomography (Micro CT; for gold standard) and CBCT scanning of the root, 1 mm of the apex was cut perpendicular to the long axis of the tooth. Each resected surface was treated with EDTA, irrigated with saline solution, and stained with methylene blue dye. The resected surface was observed with DOM and SS-OCT. This sequence was repeated three times. The number of root canals was counted and statistically evaluated. There was no significant difference in the accuracy of detecting root canals among CBCT, DOM and SS-OCT (p > 0.05, Wilcoxon test). Because SS-OCT can be used in real time during surgery, it would be a useful tool for observing resected apical root canals.

  15. Evaluation of apical extrusion of debris during ultrasonic versus rotary instrumentation

    Directory of Open Access Journals (Sweden)

    Araquam, Karolline Rodrigues

    2009-01-01

    Full Text Available Objetivo: O objetivo deste estudo foi comparar a extrusão apical de debris durante a instrumentação rotatória versus ultra-sônica de canais radiculares. Metodologia: A amostra constituiu-se de 20 pré-molares inferiores com dimensões radiculares similares. Para coleta do material extruído apicalmente durante a instrumentação endodôntica, foram fixados recipientes de resina acrílica na superfície externa das raízes. Todos os dentes tiveram os terços cervical e médio dos canais radiculares preparado com brocas Gates-Glidden. Após odontometria, a instrumentação apical foi realizada com sistema ultra-sônico ou sistema rotatório (n=10/grupo. Os recipientes de resina foram pesados antes e após a instrumentação e a diferença de peso foi considerada a quantidade de material extruído. Os dados foram analisados por teste de Mann-Whitney, ao nível de significância de 0,05. Resultados: Houve grande variabilidade de quantidade de material extruído para ambas as técnicas de instrumentação, com valores de 0,0009 a 0,0860 g. Não houve diferença estatisticamente significante entre os dois grupos (P>0. 05. Conclusão: Pode-se concluir que ambas as técnicas de instrumentação testadas, com ultra-som ou sistema rotatório, apresentaram extrusão apical de debris em quantidade semelhante

  16. Analysis of dental root apical morphology: a new method for dietary reconstructions in primates.

    Science.gov (United States)

    Hamon, NoÉmie; Emonet, Edouard-Georges; Chaimanee, Yaowalak; Guy, Franck; Tafforeau, Paul; Jaeger, Jean-Jacques

    2012-06-01

    The reconstruction of paleo-diets is an important task in the study of fossil primates. Previously, paleo-diet reconstructions were performed using different methods based on extant primate models. In particular, dental microwear or isotopic analyses provided accurate reconstructions for some fossil primates. However, there is sometimes difficult or impossible to apply these methods to fossil material. Therefore, the development of new, independent methods of diet reconstructions is crucial to improve our knowledge of primates paleobiology and paleoecology. This study aims to investigate the correlation between tooth root apical morphology and diet in primates, and its potential for paleo-diet reconstructions. Dental roots are composed of two portions: the eruptive portion with a smooth and regular surface, and the apical penetrative portion which displays an irregular and corrugated surface. Here, the angle formed by these two portions (aPE), and the ratio of penetrative portion over total root length (PPI), are calculated for each mandibular tooth root. A strong correlation between these two variables and the proportion of some food types (fruits, leaves, seeds, animal matter, and vertebrates) in diet is found, allowing the use of tooth root apical morphology as a tool for dietary reconstructions in primates. The method was then applied to the fossil hominoid Khoratpithecus piriyai, from the Late Miocene of Thailand. The paleo-diet deduced from aPE and PPI is dominated by fruits (>50%), associated with animal matter (1-25%). Leaves, vertebrates and most probably seeds were excluded from the diet of Khoratpithecus, which is consistent with previous studies.

  17. In vitro analysis of the cement film thickness of two endodontic sealers in the apical region

    Directory of Open Access Journals (Sweden)

    Garcia Lucas da Fonseca

    2009-01-01

    Full Text Available Objectives : The objective of the present study was to evaluate in vitro film thickness of two endodontic sealers containing epoxy-resin and zinc-oxide-eugenol, respectively in the apical region of the root canal system. Materials and Methods : A total of 20 palatine roots of the first maxillary molars were submitted to instrumentation up to a #40 file by the step-back technique and irrigated with 2 ml of NaOCl after each change of file. The roots were divided at random into two groups of 10 elements; one group was sealed with the Sealer 26® cement and the other one with the EndoFill® cement using the classical technique. The roots were abraded in the apical region until exposure of the filling and cut crosswise at 3 mm to obtain samples. The samples were analyzed by optic microscopy with 40X magnification. The images obtained were recorded using the Adobe Premiere 5.1® software and submitted to morphometric analysis using an integration grid that permitted the quantification of the area filled with the sealer or gutta-percha, and eventual empty spaces. Results : Statistical analysis of the data (Kruskal-Wallis test, P < 0.05 revealed no significant differences between the epoxy-resin-based cement Sealer 26® (47.85% and the zinc- oxide- eugenol-based cement EndoFill® (54.16%. Conclusion : We conclude that the two sealers tested showed a similar behavior regarding the cement film thickness in the apical region.

  18. An auxin responsive CLE gene regulates shoot apical meristem development in Arabidopsis

    Directory of Open Access Journals (Sweden)

    Hongyan eGuo

    2015-05-01

    Full Text Available Plant hormone auxin regulates most, if not all aspects of plant growth and development, including lateral root formation, organ pattering, apical dominance and tropisms. Peptide hormones are peptides with hormone activities. Some of the functions of peptide hormones in regulating plant growth and development are similar to that of auxin, however, the relationship between auxin and peptide hormones remains largely unknown. Here we report the identification of OsCLE48, a rice (Oryza sativa CLE (CLAVATA3/ENDOSPERM SURROUNDING REGION gene, as an auxin response gene, and the functional characterization of OsCLE48 in Arabidopsis and rice. OsCLE48 encodes a CLE peptide hormone that is similar to Arabidopsis CLEs. RT-PCR analysis showed that OsCLE48 was induced by exogenously application of IAA (indole-3-acetic acid, a naturally occurred auxin. Expression of integrated OsCLE48p:GUS reporter gene in transgenic Arabidopsis plants was also induced by exogenously IAA treatment. These results indicate that OsCLE48 is an auxin responsive gene. Histochemical staining showed that GUS activity was detected in all the tissue and organs of the OsCLE48p:GUS transgenic Arabidopsis plants. Expression of OsCLE48 under the control of the 35S promoter in Arabidopsis inhibited shoot apical meristem development. Expression of OsCLE48 under the control of the CLV3 native regulatory elements almost completely complemented clv3-2 mutant phenotypes, suggesting that OsCLE48 is functionally similar to CLV3. On the other hand, expression of OsCLE48 under the control of the 35S promoter in Arabidopsis has little, if any effects on root apical meristem development, and transgenic rice plants overexpressing OsCLE48 are morphologically indistinguishable from wild type plants, suggesting that the functions of some CLE peptides may not be fully conserved in Arabidopsis and rice.

  19. Changes of Root Length and Root-to-Crown Ratio after Apical Surgery

    DEFF Research Database (Denmark)

    von Arx, Thomas; Jensen, Simon S; Bornstein, Michael M

    2015-01-01

    the length of apicectomy and calculated the loss of root length and changes of RCR after apical surgery. METHODS: In a prospective clinical study, cone-beam computed tomography scans were taken preoperatively and postoperatively. From these images, the crown and root lengths of 61 roots (54 teeth in 47.......58 ± 1.43 mm (relative to the CBL). This amounted to a loss of 33.2% of clinical and 26% of anatomic root length. There was an overall significant difference between the tooth groups (P

  20. Influence of the apical enlargement size on the endotoxin level reduction of dental root canals

    Directory of Open Access Journals (Sweden)

    Ariane Cassia Salustiano Marinho

    2012-12-01

    Full Text Available Gram-negative bacteria play an essential role in endodontic infections because they have virulence factors such as endotoxin. Due to its potential cytotoxic activity, special attention has been given to the removal/neutralization of this endotoxin in the root canal system. OBJECTIVE: The aim of this study was to evaluate the influence of the apical enlargement size (AES by using rotary instruments on the endotoxin level reduction of dental root canals. MATERIAL AND METHODS: Forty root canals of the mandibular premolar teeth were used. Escherichia coli endotoxin (055: B55 was inoculated into thirty root canals. Ten teeth served as the negative control group. After the incubation period, the first endotoxin samples were collected from the root canals with a sterile/apyrogenic paper point for the analysis of the endotoxin units (EU/mL present before instrumentation (S1. Specimen instrumentation was performed with the Mtwo® rotary system in the sequence 10/.04, 15/.05, 20/.06, 25/.06, 30/.05, 35/.04 and 40/.04. To monitor the effectiveness of increasing apical enlargement on endotoxin removal, the second endotoxin samples were collected from all the root canals after instrumentation with the following instruments: #25/.06- (S2; #30/.05- (S3; # 35/.04- (S4; and #40/.04- (S5. Limulus amebocyte lysate (LAL was used to quantify the levels of endotoxin. The results were statistically compared by using repeated measures of ANOVA with post hoc Tukey testing. RESULTS: Increasing levels of endotoxin removal was achieved by large sized apical enlargement: S2 (AES #25/.06- 89.2%, S3 (AES #30/.05- 95.9%, S4 (AES #35/.04- 97.8% and S5 (AES #40/.04- 98.2%. Substantial reduction of endotoxin content was obtained in S4 and S5 compared to S2 (p<0.05, however, the root canal preparation was not able to eliminate the endotoxin. CONCLUSIONS: Under the conditions of this study, it was concluded that the reduction of endotoxin levels of the dental root canals could be

  1. An auxin responsive CLE gene regulates shoot apical meristem development in Arabidopsis.

    Science.gov (United States)

    Guo, Hongyan; Zhang, Wei; Tian, Hainan; Zheng, Kaijie; Dai, Xuemei; Liu, Shanda; Hu, Qingnan; Wang, Xianling; Liu, Bao; Wang, Shucai

    2015-01-01

    Plant hormone auxin regulates most, if not all aspects of plant growth and development, including lateral root formation, organ pattering, apical dominance, and tropisms. Peptide hormones are peptides with hormone activities. Some of the functions of peptide hormones in regulating plant growth and development are similar to that of auxin, however, the relationship between auxin and peptide hormones remains largely unknown. Here we report the identification of OsCLE48, a rice (Oryza sativa) CLE (CLAVATA3/ENDOSPERM SURROUNDING REGION) gene, as an auxin response gene, and the functional characterization of OsCLE48 in Arabidopsis and rice. OsCLE48 encodes a CLE peptide hormone that is similar to Arabidopsis CLEs. RT-PCR analysis showed that OsCLE48 was induced by exogenously application of IAA (indole-3-acetic acid), a naturally occurred auxin. Expression of integrated OsCLE48p:GUS reporter gene in transgenic Arabidopsis plants was also induced by exogenously IAA treatment. These results indicate that OsCLE48 is an auxin responsive gene. Histochemical staining showed that GUS activity was detected in all the tissue and organs of the OsCLE48p:GUS transgenic Arabidopsis plants. Expression of OsCLE48 under the control of the 35S promoter in Arabidopsis inhibited shoot apical meristem development. Expression of OsCLE48 under the control of the CLV3 native regulatory elements almost completely complemented clv3-2 mutant phenotypes, suggesting that OsCLE48 is functionally similar to CLV3. On the other hand, expression of OsCLE48 under the control of the 35S promoter in Arabidopsis has little, if any effects on root apical meristem development, and transgenic rice plants overexpressing OsCLE48 are morphologically indistinguishable from wild type plants, suggesting that the functions of some CLE peptides may not be fully conserved in Arabidopsis and rice. Taken together, our results showed that OsCLE48 is an auxin responsive peptide hormone gene, and it regulates shoot apical

  2. Rabies virus glycoprotein as a carrier for anthrax protective antigen

    International Nuclear Information System (INIS)

    Live viral vectors expressing foreign antigens have shown great promise as vaccines against viral diseases. However, safety concerns remain a major problem regarding the use of even highly attenuated viral vectors. Using the rabies virus (RV) envelope protein as a carrier molecule, we show here that inactivated RV particles can be utilized to present Bacillus anthracis protective antigen (PA) domain-4 in the viral membrane. In addition to the RV glycoprotein (G) transmembrane and cytoplasmic domains, a portion of the RV G ectodomain was required to express the chimeric RV G anthrax PA on the cell surface. The novel antigen was also efficiently incorporated into RV virions. Mice immunized with the inactivated recombinant RV virions exhibited seroconversion against both RV G and anthrax PA, and a second inoculation greatly increased these responses. These data demonstrate that a viral envelope protein can carry a bacterial protein and that a viral carrier can display whole polypeptides compared to the limited epitope presentation of previous viral systems

  3. Antigenicity analysis of Vibrio harveyi TS-628 strain

    Institute of Scientific and Technical Information of China (English)

    QIN Yingxue; WANG Jun; WANG Shifeng; YAN Qingpi

    2007-01-01

    Vibrio harveyi,the major causative agent of vibriosis,affects a diverse range of marine cultured organisms over a wide geographical area.However,reports about screening the effective antigen and research on vaccines of V.harveyi are scarce.Flagellin,lipopolysaccharide (LPS) and outer membrane proteins (OMP) are major immunogenic antigens in many Gram-negative bacteria.In this study,the flagellin,OMP and LPS of the V.harveyi TS-628 strain isolated from infected groupers were extracted and Western blot analysis was used to detect the antigenicity of these extractions.Results of the Western blot assay reveal that there are four positive flagellin bands:35 kDa,38 kDa,43 kDa,and 52 kDa,of which the 43 kDa and 52 kDa bands displayed the strongest positive reaction.There are five positive OMP bands about 35 kDa,38 kDa,43 kDa,47 kDa,and 52 kDa,of which the 43 kDa appeared to have the strongest positive reaction although the other four proteins also displayed strong reactions.However,LPS is Western blot-negative.These results indicate that the 43 kDa and 52 kDa flagellin and OMP of size 43 kDa,52 kDa can be candidates for developing vaccines against V.harveyi.

  4. Common leukocyte antigen staining of a primitive sarcoma.

    Science.gov (United States)

    McDonnell, J M; Beschorner, W E; Kuhajda, F P; deMent, S H

    1987-04-15

    A 4-year-old boy presented with symptoms of tracheal obstruction and was found to have a polypoid tracheal mass, which was studied by biopsy. Light microscopy showed a tumor composed of small cells with round to oval dark nuclei, clumped chromatin, one to two nucleoli, and small, variable amounts of indistinct pink cytoplasm. In other areas the tumor had a loose, spindle appearance, with some cells showing more elongated nuclei, and fibrillar pink cytoplasm consistent with strap cells. Cross striations were not found. Electron microscopy showed desmosomes and 7 to 10 nm cytoplasmic filaments forming dense bodies. The findings are most consistent with a primitive sarcoma, probably rhabdomyosarcoma. Immunoperoxidase with three monoclonal antibodies for common leukocyte antigen showed diffuse membraneous staining with fresh-frozen tissue. All other lymphocyte and monocyte marker studies were negative. We believe that this case of anticommon leukocyte antigen staining, a rhabdomyosarcoma, represents the first report of a false positive reaction with monoclonal antibody to common leukocyte antigen.

  5. Characterization of the basolateral membrane conductance of Necturus urinary bladder.

    Science.gov (United States)

    Demarest, J R; Finn, A L

    1987-04-01

    Necturus urinary bladders stripped of serosal muscle and connective tissue were impaled through their basolateral membranes with microelectrodes in experiments that permitted rapid changes in the ion composition of the serosal solution. The transepithelial electrical properties exhibited a marked seasonal variation that could be attributed to variations in the conductance of the shunt pathway, apical membrane selectivity, and basolateral Na+ transport. In contrast, the passive electrical properties of the basolateral membrane remained constant throughout the year. The apparent transference numbers (Ti) of the basolateral membrane for K+ and Cl- were determined from the effect on the basolateral membrane equivalent electromotive force of a sudden increase in the serosal K+ concentration from 2.5 to 50 mM/liter or a decrease in the Cl- concentration from 101 to 10 mM/liter. TK and TCl were 0.71 +/- 0.05 and 0.04 +/- 0.01, respectively. The basolateral K+ conductance could be blocked by Ba2+ (0.5 mM), Cs+ (10 mM), or Rb+ (10 mM), but was unaffected by 3,4-diaminopyridine (100 microM), decamethonium (100 microM), or tetraethylammonium (10 mM). We conclude that a highly selective K+ conductance dominates the electrical properties of the basolateral membrane and that this conductance is different from those found in nerve and muscle membranes. PMID:2438371

  6. Aluminium toxicity targets PIN2 in Arabidopsis root apices: Effects on PIN2 endocytosis, vesicular recycling,and polar auxin transport

    Institute of Scientific and Technical Information of China (English)

    SHEN Hong; HOU NingYan; Markus SCHLICHT; WAN YingLang; Stefano MANCUSO; Frantisek BALUSKA

    2008-01-01

    The most obvious symptom of AI toxicity is the inhibition of root growth.However,the mechanism of AI-inhibiting root growth remains to be elucidated.In this study,auxin transport and vesicle movement of an auxin-efflux carrier (PIN2) were investigated in Arabidopsis roots in response to AI stress.Results indicated that AI inhibited the apical transport of auxin in root tips of Arabidopsis significantly.The severe inhibition was localized in the cells of transition zone,where the concentration of auxin was only 34% that of the control.Brefeldin A (BFA),an inhibitor of vesicle transport,induced the dot-like structure of PIN2 vesicle significantly.Al decreased the size of dot-like structure of PIN2 vesicles.Re-sults of real-time RT-PCR and Western-blotting analysis showed that Al increased the transcript level of PIN2 and the accumulation of PIN2 protein in horizontal direction of plasma membrane,but decreased its distribution in endosomes,suggesting that AI inhibited the transport of PIN2 vesicles from plasma membrane to endosomes.Results of cytoskeleton-depolymering drugs indicated that it was via the pathway of disruption of actin microfilaments that AI inhibited the transport of PIN2 vesicles.Exposed to AI stress,the cells of elongation zone had less AI uptake and less transport frequency of vesicles than cells of transition zone.Taken together,our results suggested that AI inhibited root growth mainly by modulating the transport of PIN2 vesicles between plasma membrane and endosomes,thus block-ing auxin transport and root growth.

  7. Killing of Brucella antigen-sensitized macrophages by T lymphocytes in bovine brucellosis.

    Science.gov (United States)

    Wyckoff, John H; Potts, Richard D

    2007-12-15

    The present study was an investigation into the role of T lymphocytes in the killing of antigen-sensitized macrophages (M Phi) in bovine brucellosis. Following confirmation of bovine T lymphocyte cell lines derived from Brucella abortus Strain 19 vaccinated steers as antigen-specific in proliferation studies using various antigens, we adapted an apoptosis assay for evaluation of cytotoxicity by these bovine T cells against autologous monocyte-derived macrophages (MDM Phi) as target cells. Various B. abortus antigen preparations were tested including whole gamma-irradiated B. abortus bacteria (gamma BA), a soluble cytosolic protein fraction and a membrane-associated protein fraction. Both polyclonal and cloned T lymphocyte cell lines exhibited cytotoxicity against MDM Phi targets in an antigen-specific fashion. Polyclonal and cloned T lymphocyte cell lines demonstrated cytotoxic responses to varying degrees against B. abortus antigens regardless of whether the antigen used was whole nonviable bacteria, a soluble protein extract or a membrane-associated fraction of extracted bacteria. To further develop correlation of these responses to an in vivo host defense mechanism, cytotoxicity was evaluated using target cells that had been infected with live B. abortus S19 or B. abortus Strain 2308. Cytotoxic responses were also demonstrated consistently against infected targets with either strain of B. abortus although in most cases, cytotoxicity was higher against target cells sensitized with gamma BA compared to those infected with live bacteria. Cloned T lymphocyte cell lines were all CD4+, CD8(-) cells indicating that the observed cytotoxic responses were most likely due to an inflammatory Th1 response and may represent an important host defense mechanism induced by vaccination with live attenuated strains of B. abortus in cattle. PMID:17904229

  8. Noninvasive in vivo imaging reveals differences between tectorial membrane and basilar membrane traveling waves in the mouse cochlea.

    Science.gov (United States)

    Lee, Hee Yoon; Raphael, Patrick D; Park, Jesung; Ellerbee, Audrey K; Applegate, Brian E; Oghalai, John S

    2015-03-10

    Sound is encoded within the auditory portion of the inner ear, the cochlea, after propagating down its length as a traveling wave. For over half a century, vibratory measurements to study cochlear traveling waves have been made using invasive approaches such as laser Doppler vibrometry. Although these studies have provided critical information regarding the nonlinear processes within the living cochlea that increase the amplitude of vibration and sharpen frequency tuning, the data have typically been limited to point measurements of basilar membrane vibration. In addition, opening the cochlea may alter its function and affect the findings. Here we describe volumetric optical coherence tomography vibrometry, a technique that overcomes these limitations by providing depth-resolved displacement measurements at 200 kHz inside a 3D volume of tissue with picometer sensitivity. We studied the mouse cochlea by imaging noninvasively through the surrounding bone to measure sound-induced vibrations of the sensory structures in vivo, and report, to our knowledge, the first measures of tectorial membrane vibration within the unopened cochlea. We found that the tectorial membrane sustains traveling wave propagation. Compared with basilar membrane traveling waves, tectorial membrane traveling waves have larger dynamic ranges, sharper frequency tuning, and apically shifted positions of peak vibration. These findings explain discrepancies between previously published basilar membrane vibration and auditory nerve single unit data. Because the tectorial membrane directly overlies the inner hair cell stereociliary bundles, these data provide the most accurate characterization of the stimulus shaping the afferent auditory response available to date.

  9. Inhibitory activities of microalgal extracts against Epstein-Barr Virus (EBV antigen expression in lymphoblastoid cells

    Directory of Open Access Journals (Sweden)

    Koh Yih Yih

    2014-01-01

    Full Text Available The inhibitory activities of microalgal extracts against the expression of three EBV antigens, latent membrane protein (LMP1, Epstein-Barr nuclear antigen (EBNA1 and Z Epstein-Barr reactivation activator (ZEBRA were assessed by immunocytochemistry. The observation that the methanol extracts and their fractions from Ankistrodesmus convolutus, Synechococcus elongatus and Spirulina platensis exhibited inhibitory activity against EBV proteins in three Burkitt’s lymphoma cell lines at concentrations as low as 20 μg/ml suggests that microalgae could be a potential source of antiviral compounds against EBV.

  10. Membrane potentials and intracellular Cl- activity of toad skin epithelium in relation to activation and deactivation of the transepithelial Cl- conductance

    DEFF Research Database (Denmark)

    Willumsen, N J; Larsen, Erik Hviid

    1986-01-01

    , but was independent of whether the external bath was NaCl-Ringer's, NaCl-Ringer's with amiloride, KCl-Ringer's or choline Cl-Ringer's. To partition the routes of the conductive Cl- ion flow, we measured in the isolated epithelium with double-barrelled microelectrodes apical membrane potential. Va, and intracellular...

  11. Omniphobic Membrane for Robust Membrane Distillation

    Energy Technology Data Exchange (ETDEWEB)

    Lin, SH; Nejati, S; Boo, C; Hu, YX; Osuji, CO; Ehmelech, M

    2014-11-01

    In this work, we fabricate an omniphobic microporous membrane for membrane distillation (MD) by modifying a hydrophilic glass fiber membrane with silica nanoparticles followed by surface fluorination and polymer coating. The modified glass fiber membrane exhibits an anti-wetting property not only against water but also against low surface tension organic solvents that easily wet a hydrophobic polytetrafluoroethylene (PTFE) membrane that is commonly used in MD applications. By comparing the performance of the PTFE and omniphobic membranes in direct contact MD experiments in the presence of a surfactant (sodium dodecyl sulfate, SDS), we show that SDS wets the hydrophobic PTFE membrane but not the omniphobic membrane. Our results suggest that omniphobic membranes are critical for MD applications with feed waters containing surface active species, such as oil and gas produced water, to prevent membrane pore wetting.

  12. Endodontic management of immature teeth with spontaneous apical closure and periapical lesions: case series and review of the literature.

    Science.gov (United States)

    Çalışkan, Mehmet Kemal; Kaval, Mehmet Emin

    2015-08-01

    Spontaneous apical closure in non-vital immature teeth has been rarely encountered and outcome of non-surgical endodontic treatment of related teeth associated with periapical lesions has not yet been adequately elucidated. The aim of this article was to report endodontic management of spontaneous apical closure of infected untreated immature teeth with periapical lesions and to review previously proposed mechanisms for the development of spontaneous hard tissue barrier. Three patients were referred at different time intervals to the endodontic clinic for treatment of their maxillary anterior incisors with acute or chronic apical periodontitis. Dental histories indicated that related teeth had been subjected to trauma approximately 12-18 years previously. Radiographically, the involved teeth exhibited incomplete root formation with spontaneous apical closure and were associated with an apical radiolucency. After biomechanical preparation, calcium hydroxide paste was applied and was changed once or twice within 3 months. All canals were then filled with gutta-percha and AH Plus and the follow-up period was 16-50 months; both clinical and radiographic examinations revealed adequate function, the absence of clinical symptoms and significant healing of the periapical radiolucency.

  13. Fragment Propagation and Colonization Ability Enhanced and Varied at Node Level after Escaping from Apical Dominance in Submerged Macrophytes

    Institute of Scientific and Technical Information of China (English)

    Jinhui Jiang; Shuqing An; Changfang Zhou; Baohua Guan; Zhiyi Sun; Ying Cai; Fude Liu

    2009-01-01

    Aquatic plants develop strong fragment propagation and colonization ability to endure the natural disturbances. However,detailed research of ability to endure the natural disturbances has been lacking to date. Therefore, reproduction (shoot) and colonization (root) of shoot fragments of Potamogeton crispus L. with or without apices were investigated for the effect of apical dominance, and the growth of decapitated shoot fragments at three lengths (2, 4, 6cm) was compared. Meanwhile,fragment propagation at levels of bud position was studied for bud position effect after escaping from apical dominance.The results showed significant increases occurred in the outgrowth of lateral branches on fragments decapitated compared with the fragments with apices, implying that apical dominance exists. Different lengths of fragments showed little difference in biomass allocations, but significant differences were noted in their propagation. Meanwhile, the effect of bud position was verified, due to the significant difference of average reproduction per node among the three length groups. Thus, the present study has made progress in the current understanding of aquatic plant dispersion among natural systems and contributes to improve methods of in vitro propagation for re-implantation purposes.

  14. Surgical closure of apical multiple muscular septal defects via right ventriculotomy using a single patch with intermediate fixings

    Institute of Scientific and Technical Information of China (English)

    LI Gang; WANG Qiang; LIU Ying-long; XU Yu-lin; GUO Yan

    2013-01-01

    Background Multiple apical muscular ventricular septal defects (VSDs) remain a challenge for surgeons because of their anatomical features.We used single patch with intermediate fixations to repair multiple apical muscular VSDs through right ventriculotomy.Methods We analysed the data of 16 children (median age 8 months,range 2 months to 144 months) with multiple apical muscular VSDs who underwent a single patch technique via apical right ventriculotomy.Perioperative data were collected and analysed,and the patients were followed up for three months to 66 months (median,46 months) to investigate the outcomes.Results All patients recovered from cardiopulmonary bypass easily with median of cardiopulmonary bypass time 87 minutes and of aortic crossclamp time 53 minutes.No surgically related death occurred and no patient required reoperation.One patient died of pseudomonas pyocyanea infection on day 11 postoperatively.Residual shunt happened in one patient with a diameter of 2 mm and spontaneously closed in 12 months after operation.Two patients presented slightly reduced right ventricular volume and apical hypokinesia postoperatively and recovered 24 and 36 months later respectively.No other complication was found during the three months to 66 months follow-up.Conclusion Our experiences using a single patch technique with intermediate fixations via right ventriculotomy in the management of multiple muscular VSDs had favourable outcomes.

  15. Ecology of the microbiome of the infected root canal system: a comparison between apical and coronal root segments

    Science.gov (United States)

    Özok, A.R.; Persoon, I.F.; Huse, S.M.; Keijser, B.J.F.; Wesselink, P.R.; Crielaard, W.; Zaura, E.

    2016-01-01

    Aim To evaluate the microbial ecology of the coronal and apical segments of infected root canal systems using a complete sampling technique and next-generation sequencing. Methodology The roots of 23 extracted teeth with apical periodontitis were sectioned in half, horizontally, and cryo-pulverized. Bacterial communities were profiled using tagged 454 pyrosequencing of the 16S rDNA hypervariable V5–V6 region. Results The sequences were classified into 606 taxa (species or higher taxon), representing 24 bacterial phyla or candidate divisions and one archaeal phylum. Proteobacteria were more abundant in the apical samples (proots compared to their coronal counterparts. Conclusions Endodontic infections are more complex than reported previously. The apical part of the root canal system drives the selection of a more diverse and more anaerobe community than the coronal part. The presence of a distinct ecological niche in the apical region explains the difficulty of eradication of the infection, and emphasizes the need that new treatment approaches should be developed. PMID:22251411

  16. Anatomy and histology of apical support: a literature review concerning cardinal and uterosacral ligaments.

    Science.gov (United States)

    Ramanah, Rajeev; Berger, Mitchell B; Parratte, Bernard M; DeLancey, John O L

    2012-11-01

    The objective of this work was to collect and summarize relevant literature on the anatomy, histology, and imaging of apical support of the upper vagina and the uterus provided by the cardinal (CL) and uterosacral (USL) ligaments. A literature search in English, French, and German languages was carried out with the keywords apical support, cardinal ligament, transverse cervical ligament, Mackenrodt ligament, parametrium, paracervix, retinaculum uteri, web, uterosacral ligament, and sacrouterine ligament in the PubMed database. Other relevant journal and textbook articles were sought by retrieving references cited in previous PubMed articles. Fifty references were examined in peer-reviewed journals and textbooks. The USL extends from the S2 to the S4 vertebra region to the dorsal margin of the uterine cervix and/or to the upper third of the posterior vaginal wall. It has a superficial and deep component. Autonomous nerve fibers are a major constituent of the deep USL. CL is defined as a perivascular sheath with a proximal insertion around the origin of the internal iliac artery and a distal insertion on the cervix and/or vagina. It is divided into a cranial (vascular) and a caudal (neural) portions. Histologically, it contains mainly vessels, with no distinct band of connective tissue. Both the deep USL and the caudal CL are closely related to the inferior hypogastric plexus. USL and CL are visceral ligaments, with mesentery-like structures containing vessels, nerves, connective tissue, and adipose tissue.

  17. A survey of small RNA population during FR-induced apical hook opening

    Directory of Open Access Journals (Sweden)

    Ying eLi

    2014-04-01

    Full Text Available Photomorphogenesis is a mechanism employed by plants to regulate their architecture and developmental program in response to light conditions. As they emerge into light for the first time, dark-grown seedlings employ a rapid and finely-controlled photomorphogenic signaling network. Small RNAs have increasingly been revealed to play an important role in regulating multiple aspects of plant development, by modulating the stability of mRNAs. The rapid alteration of the mRNA transcriptome is a known hallmark of the de-etiolation response, thus we investigated the small RNA transcriptome during this process in specific seedling tissues. Here we describe a survey of the small RNA expression profile in four tissues of etiolated soybean seedlings, the cotyledons, hypocotyl and the convex and concave sides of the apical hook. We also investigate how this profile responds to a one-hour far-red light treatment. Our data suggests that miRNAs show a different global profile between these tissues and treatments, suggesting a possible role for tissue- and treatment-specific expression in the differential morphology of the seedling on de-etiolation. Further evidence for the role of miRNA in light-regulated development is given by the de-etiolation responses of a hypomorphic ago1 mutant, which displays reduced and delayed photomorphogenic responses in apical hook and cotyledon angle to far-red light.

  18. Multiciliated cell basal bodies align in stereotypical patterns coordinated by the apical cytoskeleton.

    Science.gov (United States)

    Herawati, Elisa; Taniguchi, Daisuke; Kanoh, Hatsuho; Tateishi, Kazuhiro; Ishihara, Shuji; Tsukita, Sachiko

    2016-08-29

    Multiciliated cells (MCCs) promote fluid flow through coordinated ciliary beating, which requires properly organized basal bodies (BBs). Airway MCCs have large numbers of BBs, which are uniformly oriented and, as we show here, align linearly. The mechanism for BB alignment is unexplored. To study this mechanism, we developed a long-term and high-resolution live-imaging system and used it to observe green fluorescent protein-centrin2-labeled BBs in cultured mouse tracheal MCCs. During MCC differentiation, the BB array adopted four stereotypical patterns, from a clustering "floret" pattern to the linear "alignment." This alignment process was correlated with BB orientations, revealed by double immunostaining for BBs and their asymmetrically associated basal feet (BF). The BB alignment was disrupted by disturbing apical microtubules with nocodazole and by a BF-depleting Odf2 mutation. We constructed a theoretical model, which indicated that the apical cytoskeleton, acting like a viscoelastic fluid, provides a self-organizing mechanism in tracheal MCCs to align BBs linearly for mucociliary transport. PMID:27573463

  19. Regulated Crb accumulation controls apical constriction and invagination in Drosophila tracheal cells.

    Science.gov (United States)

    Letizia, Annalisa; Sotillos, Sol; Campuzano, Sonsoles; Llimargas, Marta

    2011-01-15

    Many epithelial tissues undergo extensive remodelling during morphogenesis. How their epithelial features, such as apicobasal polarity or adhesion, are maintained and remodelled and how adhesion and polarity proteins contribute to morphogenesis are two important questions in development. Here, we approach these issues by investigating the role of the apical determinant protein Crumbs (Crb) during the morphogenesis of the embryonic Drosophila tracheal system. Crb accumulates differentially throughout tracheal development and is required for different tracheal events. The earliest requirement for Crb is for tracheal invagination, which is preceded by an enhanced accumulation of Crb in the invagination domain. There, Crb, acting in parallel with the epidermal growth factor receptor (Egfr) pathway, is required for tracheal cell apical constriction and for organising an actomyosin complex, which we propose is mediated by Crb recruitment of moesin (Moe). The ability of a Crb isoform unable to rescue polarity in crb mutants to otherwise rescue their invagination phenotype, and the converse inability of a FERM-binding domain mutant Crb to rescue faulty invagination, support our hypothesis that it is the absence of Crb-dependent Moe enrichment, and not the polarity defect, that mainly underlies the crb invagination phenotype. This hypothesis is supported by the phenotype of lethal giant larvae (lgl); crb double mutants. These results unveil a link between Crb and the organisation of the actin cytoskeleton during morphogenesis. PMID:21172808

  20. Auxin controls local cytokinin biosynthesis in the nodal stem in apical dominance.

    Science.gov (United States)

    Tanaka, Mina; Takei, Kentaro; Kojima, Mikiko; Sakakibara, Hitoshi; Mori, Hitoshi

    2006-03-01

    In intact plants, the shoot apex grows predominantly and inhibits outgrowth of axillary buds. After decapitation of the shoot apex, outgrowth of axillary buds begins. This phenomenon is called an apical dominance. Although the involvement of auxin, which represses outgrowth of axillary buds, and cytokinin (CK), which promotes outgrowth of axillary buds, has been proposed, little is known about the underlying molecular mechanisms. In the present study, we demonstrated that auxin negatively regulates local CK biosynthesis in the nodal stem by controlling the expression level of the pea (Pisum sativum L.) gene adenosine phosphate-isopentenyltransferase (PsIPT), which encodes a key enzyme in CK biosynthesis. Before decapitation, PsIPT1 and PsIPT2 transcripts were undetectable; after decapitation, they were markedly induced in the nodal stem along with accumulation of CK. Expression of PsIPT was repressed by the application of indole-3-acetic acid (IAA). In excised nodal stem, PsIPT expression and CK levels also increased under IAA-free conditions. Furthermore, beta-glucuronidase expression, under the control of the PsIPT2 promoter region in transgenic Arabidopsis, was repressed by an IAA. Our results indicate that in apical dominance one role of auxin is to repress local biosynthesis of CK in the nodal stem and that, after decapitation, CKs, which are thought to be derived from the roots, are locally biosynthesized in the nodal stem rather than in the roots. PMID:16507092

  1. Postoperative left ventricular apical ballooning: Transient Takotsubo cardiomyopathy following orthotopic liver transplantation

    Science.gov (United States)

    Bedanova, Helena; Orban, Marek; Nemec, Petr

    2013-01-01

    Patient: Female, 51 Final Diagnosis: Takotsubo cardiomyopathy Symptoms: — Medication: — Clinical Procedure: — Specialty: Cardiology • Transplantology Objective: Rare disease Background: Left ventricular apical ballooning syndrome (LVAB), also known as Takotsubo cardiomyopathy, is a cardiac syndrome characterized by transient left ventricular dysfunction in the absence of obstructive atherosclerotic coronary artery disease. An episode of emotional stress, typically in female patients, is believed to precede and trigger the development of this syndrome. Case Report: We report a case of Takotsubo cardiomyopathy that developed after orthotopic liver transplantation in a 51-year-old woman. On D2 (day 2) the patient had severe hemodynamic compromise. Echocardiography showed systolic dysfunction of the left ventricle (LV), with ejection fraction (EF) of 20% and anteroapical akinesis and ballooning of the apical 2/3 of the LV. Troponin T was elevated but other markers of myocardial necrosis were negative, as was coronary angiography. From D7 onward, there was an improvement in the hemodynamics in conjunction with a gradual increase of LV EF. The patient was dismissed from the hospital on D30 with signs of normal cardiac function and LV motion and EF of 50%. Liver function was also excellent. Conclusions: Every major operation, including liver transplantation, is associated with emotional stress for the patient. Therefore, it is necessary to consider Takotsubo cardiomyopathy in the differential diagnosis of heart failure developing early after LT, and clinicians should subsequently use adequate diagnostic and therapeutic measures. PMID:24298303

  2. SHA1, a novel RING finger protein, functions in shoot apical meristem maintenance in Arabidopsis.

    Science.gov (United States)

    Sonoda, Yutaka; Yao, Shan-Guo; Sako, Kaori; Sato, Takeo; Kato, Wataru; Ohto, Masa-aki; Ichikawa, Takanari; Matsui, Minami; Yamaguchi, Junji; Ikeda, Akira

    2007-05-01

    Post-embryonic plant growth is dependent on a functional shoot apical meristem (SAM) that provides cells for continuous development of new aerial organs. However, how the SAM is dynamically maintained during vegetative development remains largely unclear. We report here the characterization of a new SAM maintenance mutant, sha1-1 (shoot apical meristem arrest 1-1), that shows a primary SAM-deficient phenotype at the adult stage. The SHA1 gene encodes a novel RING finger protein, and is expressed most intensely in the shoot apex. We show that, in the sha1-1 mutant, the primary SAM develops normally during the juvenile vegetative stage, but cell layer structure becomes disorganized after entering the adult vegetative stage, resulting in a dysfunctional SAM that cannot initiate floral primordia. The sha1-1 SAM terminates completely at the stage when the wild-type begins to bolt, producing adult plants with a primary inflorescence-deficient phenotype. These observations indicate that SHA1, a putative E3 ligase, is required for post-embryonic SAM maintenance by controlling proper cellular organization. PMID:17461786

  3. RNG1 is a late marker of the apical polar ring in Toxoplasma gondii.

    Science.gov (United States)

    Tran, Johnson Q; de Leon, Jessica C; Li, Catherine; Huynh, My-Hang; Beatty, Wandy; Morrissette, Naomi S

    2010-09-01

    The asexually proliferating stages of apicomplexan parasites cause acute symptoms of diseases such as malaria, cryptosporidiosis and toxoplasmosis. These stages are characterized by the presence of two independent microtubule organizing centers (MTOCs). Centrioles are found at the poles of the intranuclear spindle. The apical polar ring (APR), a MTOC unique to apicomplexans, organizes subpellicular microtubules which impose cell shape and apical polarity on these protozoa. Here we describe the characteristics of a novel protein that localizes to the APR of Toxoplasma gondii which we have named ring-1 (RNG1). There are related RNG1 proteins in Neospora caninum and Sarcocystis neurona but no obvious homologs in Plasmodium spp., Cryptosporidium spp. or Babesia spp. RNG1 is a small, low-complexity, detergent-insoluble protein that assembles at the APR very late in the process of daughter parasite replication. We were unable to knock-out the RNG1 gene, suggesting that its gene product is essential. Tagged RNG1 lines have also allowed us to visualize the APR during growth of Toxoplasma in the microtubule-disrupting drug oryzalin. Oryzalin inhibits nuclear division and cytokinesis although Toxoplasma growth continues, and similar to earlier observations of unchecked centriole duplication in oryzalin-treated parasites, the APR continues to duplicate during aberrant parasite growth. PMID:20658557

  4. Effect of diode laser irradiation on the apical sealing of MTA retrofillings Efeito da irradiação de laser de diodo no selamento apical em retrobturações com MTA

    Directory of Open Access Journals (Sweden)

    Eliana Barbosa de Souza

    2006-09-01

    Full Text Available Apical sealing is essential for the success of paraendodontic surgery, so any procedure that may favor an adequate sealing of the apical remainder should be performed. The purpose of this study was to evaluate the influence of diode laser irradiation on the apical sealing of root-end cavities with MTA retrofillings. Root canals in twenty extracted human teeth were shaped with K-files and filled with gutta-percha. The apexes were cut off and root-end preparations were performed. The roots were divided randomly in 2 groups. Group 1 (ten specimens was retrofilled with MTA. Group 2 was irradiated with diode laser, with 1 W for 20 seconds, on the apical surface and root end cavity before retrofilling with MTA. The specimens had their external surfaces impermeabilized with cyanoacrylate, except for the apical surface, and were then immersed in 1% rhodamine B dye for 72 h and placed in plaster stone. After that, the specimens were submitted to longitudinal abrasion until half of the root remained. The linear dye leakage was observed in these mid-roots between the root canal wall and retrofilling. The linear dye leakage was measured with Image Lab software, and the results were statistically analyzed with Student's t test. There were no statistically significant differences between the two groups (p > 0.05. The diode laser irradiation did not improve the apical sealing of MTA retrofillings under the conditions of this in vitro study.O selamento apical é fundamental para o sucesso da cirurgia parendodôntica. Assim, procedimentos que melhorem o selamento do remanescente apical devem ser utilizados. O objetivo deste estudo foi verificar se a irradiação de laser de diodo poderia aumentar o selamento apical em cavidades retrógradas obturadas com MTA. Foram utilizadas 20 raízes de dentes humanos extraídos que, após preparo com lima tipo K, tiveram seus canais obturados com guta-percha. Os ápices foram cortados e sofreram preparo de cavidades retr

  5. Posterior midgut epithelial cells differ in their organization of the membrane skeleton from other drosophila epithelia.

    Science.gov (United States)

    Baumann, O

    2001-11-01

    In epithelial cells, the various components of the membrane skeleton are segregated within specialized subregions of the plasma membrane, thus contributing to the development and stabilization of cell surface polarity. It has previously been shown that, in various Drosophila epithelia, the membrane skeleton components ankyrin and alphabeta-spectrin reside at the lateral surface, whereas alphabeta(H)-spectrin is restricted to the apical domain. By use of confocal immunofluorescence microscopy, the present study characterizes the membrane skeleton of epithelial cells in the posterior midgut, leading to a number of unexpected results. First, ankyrin and alphabeta-spectrin are not detected on the entire lateral surface but appear to be restricted to the apicolateral area, codistributing with fasciclin III at smooth septate junctions. The presumptive ankyrin-binding proteins neuroglian and Na(+),K(+)-ATPase, however, do not colocalize with ankyrin. Second, alphabeta(H)-spectrin is enriched at the apical domain but is also present in lower amounts on the entire lateral surface, colocalizing apicolaterally with ankyrin/alphabeta-spectrin. Finally, despite the absence of zonulae adherentes, F-actin, beta(H)-spectrin, and nonmuscle myosin-II are enriched in the midlateral region. Thus, the model established for the organization of the membrane skeleton in Drosophila epithelia does not hold for the posterior midgut, and there is quite some variability between the different epithelia with respect to the organization of the membrane skeleton.

  6. The apical leakage of mineral trioxide aggregate as the retrograde filling material with various mixing agents

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    Ema Mulyawati

    2010-06-01

    Full Text Available Background: Mineral trioxide aggregate (MTA is relatively considered as a new material in endodontic. It even has been used as retrograde filling material due to its biocompatibility, antibacterial effect, sealing ability and anti-moist effect. Some materials have been used as mixing agent to achieve an appropiate setting of MTA. Purpose: The aim of this study is to investigate the effect of the mixing agents of MTA towards the apical leakage when they are used together as retrograde filling materials. Method: The samples of this research consist of 30 human extracted upper central incisors. First, the crown of each tooth is sectioned. The root canals are prepared by using the conventional technique and then are obturated with gutta percha. After cutting the root apex, 2 mm from apical, class 1 cavities are prepared by using fissure bur with the depth of 3 mm. The samples then are divided into 3 groups with 10 teeth for each. Group I uses aquabidest as mixing agent of MTA (MTA-aquabidest, group II uses saline (MTA-saline, while group III uses 0.12% chlorhexidine (MTA-chlorhexidine. The apex of each group then is filled with the mixing MTA determined already. Afterwards, clearing method is used to evaluate the apical leakage. The apical leakage actually is determined by measuring the depth of methylene blue penetration with stereomicroscope. The statictical analyses of the linear dye penetration then are performed with analysis of varians ANOVA. Result: The dye penetration for both MTA-aquadest and MTA-saline groups indicates the lowest penetration, and there is even a significant difference compared with MTA-0.12% chlorhexidine group (p<0.005. Conclusion: It can be concluded that aquabidest and saline as mixing agents of MTA produce less apical leakage compared with 0.12% chlorhexidine.Latar belakang: Mineral trioxide aggregate (MTA merupakan bahan yang relatif baru dalam bidang endodontik. Bahan tersebut diindikasikan sebagai bahan pengisi

  7. Exposed and concealed antigens as vaccine targets for controlling ticks and tick-borne diseases.

    Science.gov (United States)

    Nuttall, P A; Trimnell, A R; Kazimirova, M; Labuda, M

    2006-04-01

    Tick vaccines derived from Bm86, a midgut membrane-bound protein of the cattle tick, Boophilus microplus, are currently the only commercially available ectoparasite vaccines. Despite its introduction to the market in 1994, and the recognized need for alternatives to chemical pesticides, progress in developing effective antitick vaccines (and ectoparasite vaccines in general) is slow. The primary rate-limiting step is the identification of suitable antigenic targets for vaccine development. Two sources of candidate vaccine antigens have been identified: 'exposed' antigens that are secreted in tick saliva during attachment and feeding on a host and 'concealed' antigens that are normally hidden from the host. Recently, a third group of antigens has been distinguished that combines the properties of both exposed and concealed antigens. This latter group offers the prospect of a broad-spectrum vaccine effective against both adults and immature stages of a wide variety of tick species. It also shows transmission-blocking and protective activity against a tick-borne pathogen. With the proliferation of molecular techniques and their application to vaccine development, there are high hopes for new and effective antitick vaccines that also control tick-borne diseases. PMID:16542317

  8. Par1b induces asymmetric inheritance of plasma membrane domains via LGN-dependent mitotic spindle orientation in proliferating hepatocytes.

    Directory of Open Access Journals (Sweden)

    Christiaan L Slim

    2013-12-01

    Full Text Available The development and maintenance of polarized epithelial tissue requires a tightly controlled orientation of mitotic cell division relative to the apical polarity axis. Hepatocytes display a unique polarized architecture. We demonstrate that mitotic hepatocytes asymmetrically segregate their apical plasma membrane domain to the nascent daughter cells. The non-polarized nascent daughter cell can form a de novo apical domain with its new neighbor. This asymmetric segregation of apical domains is facilitated by a geometrically distinct "apicolateral" subdomain of the lateral surface present in hepatocytes. The polarity protein partitioning-defective 1/microtubule-affinity regulating kinase 2 (Par1b/MARK2 translates this positional landmark to cortical polarity by promoting the apicolateral accumulation of Leu-Gly-Asn repeat-enriched protein (LGN and the capture of nuclear mitotic apparatus protein (NuMA-positive astral microtubules to orientate the mitotic spindle. Proliferating hepatocytes thus display an asymmetric inheritance of their apical domains via a mechanism that involves Par1b and LGN, which we postulate serves the unique tissue architecture of the developing liver parenchyma.

  9. Engineering the chloroplast targeted malarial vaccine antigens in Chlamydomonas starch granules.

    Directory of Open Access Journals (Sweden)

    David Dauvillée

    Full Text Available BACKGROUND: Malaria, an Anopheles-borne parasitic disease, remains a major global health problem causing illness and death that disproportionately affects developing countries. Despite the incidence of malaria, which remains one of the most severe infections of human populations, there is no licensed vaccine against this life-threatening disease. In this context, we decided to explore the expression of Plasmodium vaccine antigens fused to the granule bound starch synthase (GBSS, the major protein associated to the starch matrix in all starch-accumulating plants and algae such as Chlamydomonas reinhardtii. METHODS AND FINDINGS: We describe the development of genetically engineered starch granules containing plasmodial vaccine candidate antigens produced in the unicellular green algae Chlamydomonas reinhardtii. We show that the C-terminal domains of proteins from the rodent Plasmodium species, Plasmodium berghei Apical Major Antigen AMA1, or Major Surface Protein MSP1 fused to the algal granule bound starch synthase (GBSS are efficiently expressed and bound to the polysaccharide matrix. Mice were either immunized intraperitoneally with the engineered starch particles and Freund adjuvant, or fed with the engineered particles co-delivered with the mucosal adjuvant, and challenged intraperitoneally with a lethal inoculum of P. Berghei. Both experimental strategies led to a significantly reduced parasitemia with an extension of life span including complete cure for intraperitoneal delivery as assessed by negative blood thin smears. In the case of the starch bound P. falciparum GBSS-MSP1 fusion protein, the immune sera or purified immunoglobulin G of mice immunized with the corresponding starch strongly inhibited in vitro the intra-erythrocytic asexual development of the most human deadly plasmodial species. CONCLUSION: This novel system paves the way for the production of clinically relevant plasmodial antigens as algal starch-based particles

  10. Fusion of liposomes with the plasma membrane of epithelial cells: Fate of incorporated lipids as followed by freeze fracture and autoradiography of plastic sections

    OpenAIRE

    Knoll, G.; Burger, K.N.J.; Bron, R.; van Meer, G.; Verkleij, A. J.

    1988-01-01

    The fusion of liposomes with the plasma membrane of influenza virus- infected monolayers of an epithelial cell line, Madin-Darby canine kidney cells (van Meer et al., 1985. Biochemistry. 24:3593-3602), has been analyzed by morphological techniques. The distribution of liposomal lipids over the apical and basolateral plasma membrane domains after fusion was assessed by autoradiography of liposomal [3H]dipalmitoylphosphatidylcholine after rapid freezing or chemical fixation and further processi...

  11. Delayed-enhancement MRI of apical hypertrophic cardiomyopathy: assessment of the intramural distribution and comparison with clinical symptoms, ventricular arrhythmias, and cine MRI

    Energy Technology Data Exchange (ETDEWEB)

    Amano, Yasuo; Fukushima, Yoshimitsu; Kumita, Shinichiro (Dept. of Radiology, Nippon Medical School, Tokyo (Japan)), email: yas-amano@nifty.com; Takayama, Morimasa (Dept. of Cardiology, Sakakibara Heart Inst., Tokyo (Japan)); Kitamura, Mitsunobu (Coronary Care Unit, Chiba-Hokuso Hospital of Nippon Medical School, Chiba (Japan))

    2011-07-15

    Background: Hypertrophic cardiomyopathy (HCM) is reported to show patchy midwall myocardial hyper enhancement on delayed-enhancement magnetic resonance imaging (DE-MRI). The intramural distribution of myocardial hyper enhancement and its correlation with clinical symptoms, ventricular arrhythmias, and cardiac function have not been described for symptomatic apical HCM. Purpose: To evaluate the features and significance of myocardial hyper enhancement on DE-MRI in symptomatic apical HCM. Material and Methods: Thirteen patients with symptomatic apical HCM and their 65 apical segments were investigated. Myocardial hyper enhancement and regional and global functional parameters were determined with MRI. We investigated the intramural distribution and frequencies of this myocardial hyper enhancement and compared them with the patients' clinical symptoms, the presence of ventricular arrhythmias, and cine MRI. Results: Eight (61.5%) patients with symptomatic apical HCM displayed apical myocardial hyper enhancement, and 22 (33.8%) of the 65 apical segments examined showed myocardial hyper enhancement. Of the myocardial hyper enhancement observed, 81.8% showed a subendocardial pattern.The Hyperenhanced apical myocardium had a lower percentage of systolic myocardial thickening, and was associated with serious symptoms (e.g. syncope) and ventricular arrhythmias. Conclusion: Patients with symptomatic apical HCM showed myocardial hyper enhancement involving the subendocardial layer, which might be related to regional systolic dysfunction, serious clinical symptoms, and ventricular arrhythmias

  12. Evaluation of apical deviation in root canals instrumented with K3 and ProTaper systems Avaliação do desvio apical em canais instrumentados pelos sistemas K3 e ProTaper

    Directory of Open Access Journals (Sweden)

    Mariana Diniz Bisi dos Santos

    2006-12-01

    Full Text Available OBJECTIVES: this study evaluated the apical deviation of curved root canals instrumented with K3 and ProTaper systems. MATERIAL AND METHODS: twenty root canals of human maxillary and mandibular first molars were employed, which were divided into 2 groups: group A (10 teeth was instrumented with the K3 system, and group B (10 teeth with the ProTaper system. Evaluation of deviation was performed by double radiographic exposure. Radiographs were achieved before and after instrumentation, with 0.3-second, thus allowing superimposition of images. Three-dimensional computerized tomograph was performed in 3 specimens in each group, as an additional means to evaluate the apical deviation. RESULTS: were evaluated by the parametric test Student-Newman-Keuls at 5%, which did not reveal significance between groups concerning the apical deviation. The results of computerized tomograph images demonstrated that the larger deviation of the root canal occurred at the distolingual area for both systems. CONCLUSIONS: both techniques produced a mild apical deviation. Computerized microtomography was shown to be accurate for experimental endodontics studies.OBJETIVOS: avaliar o desvio apical de canais radiculares curvos instrumentados pelos sistemas K3 e ProTaper. MATERIAIS E MÉTODOS: foram utilizados 20 canais radiculares de primeiros molares superiores e inferiores humanos, os quais foram divididos em 2 grupos, o grupo A (10 dentes foi instrumentado pelo sistema K3 e o grupo B (10 dentes com o sistema ProTaper, a forma de avaliação do desvio foi a dupla exposição radiográfica. As tomadas radiográficas foram feitas antes e após a instrumentação, com um tempo de 0,3 segundos em cada exposição, ocorrendo assim, sobreposição das imagens. Em 3 espécimes, de cada grupo, foi realizada a tomografia computadorizada tridimensional, como forma adicional de avaliar o desvio apical . Os resultados obtidos foram avaliados pelo teste paramétrico Student

  13. Mid-ventricular obstructive hypertrophic cardiomyopathy with apical aneurysm and sustained ventricular tachycardia: a case report and literature review

    Institute of Scientific and Technical Information of China (English)

    GAO Xiao-jin; KANG Lian-ming; ZHANG Jian; DOU Ke-fei; YUAN Jian-song; YANG Yue-jin

    2011-01-01

    The case is a 54-year-old man with hypertrophic cardiomyopathy, mid-ventricular obstruction, apical aneurysm, and recurrence sustained monomorphic ventricular tachycardia (VT). A coronary angiogram revealed myocardial bridging located in the middle of the left anterior descending coronary artery (LAD), and the left ventriculogram showed an hour-glass appearance of the left ventricular cavity. There was a significant pressure gradient of 60 mmHg across the mid-ventricular obliteration at rest. A successful myectomy of the inappropriate hypertrophy myocardium and excision of the apical aneurysm were performed. Pathologic analysis demonstrated fibrosis in the apical aneurysm and thickened and narrowed vessels in the adjacent area. During the follow-up of eighteen months, the patient remained clinically stable and free from arrhythmic recurrence.

  14. Antigen Incorporation on Cryptosporidium parvum Oocyst Walls

    OpenAIRE

    Entrala Emilio; Sbihi Younes; Sánchez-Moreno Manuel; Mascaró Carmen

    2001-01-01

    Cryptosporidium parvum oocysts are the infective stages responsible for transmission and survival of the organism in the environment. In the present work we show that the oocyst wall, far from being a static structure, is able to incorporate antigens by a mechanism involving vesicle fusion with the wall, and the incorporation of the antigen to the outer oocyst wall. Using immunoelectron microscopy we show that the antigen recognized by a monoclonal antibody used for diagnosis of cryptosporidi...

  15. Histocompatibility antigens in coal miners with pneumoconiosis.

    OpenAIRE

    Soutar, C A; Coutts, I.; Parkes, W R; Dodi, I. A.; Gauld, S; Castro, J E; Turner-Warwick, M

    1983-01-01

    Twenty-five histocompatibility antigens have been measured in 100 coal miners with pneumoconiosis attending a pneumoconiosis medical panel and the results compared with a panel of 200 normal volunteers not exposed to dust. Chest radiographs were read independently by three readers according to the ILO U/C classification. On a combined score, 40 men were thought to have simple pneumoconiosis and 60 men complicated pneumoconiosis. The number of antigens tested and associations between antigens ...

  16. Secretion, interaction and assembly of two O-glycosylated cell wall antigens from Candida albicans.

    Science.gov (United States)

    Pavia, J; Aguado, C; Mormeneo, S; Sentandreu, R

    2001-07-01

    The mechanisms of incorporation of two antigens have been determined using a monoclonal antibody (3A10) raised against the material released from the mycelial cell wall by zymolyase digestion and retained on a concanavalin A column. One of the hybridomas secreted an IgG that reacted with two bands in Western blots. Indirect immunofluorescence showed that the antigens were located on the surfaces of mycelial cells, but within the cell walls of yeasts. These antigens were detected in a membrane preparation, in the SDS-soluble material and in the material released by a 1,3-beta-glucanase and chitinase from the cell walls of yeast and mycelial cells. In the latter three samples, an additional high-molecular-mass, highly polydispersed band was also detected. Beta-elimination of each fraction resulted in the disappearance of all antigen bands, suggesting that they are highly O-glycosylated. In addition, the electrophoretic mobility of the high-molecular-mass, highly polydispersed bands increased after digestion with endoglycosidase H, indicating that they are also N-glycosylated. New antigen bands were released when remnants of the cell walls extracted with 1,3-beta-glucanase or chitinase were digested with chitinase or 1,3-beta-glucanase. These results are consistent with the notion that, after secretion, parts of the O-glycosylated antigen molecules are transferred to an N-glycosylated protein(s). This molecular complex, as well as the remaining original 70 and 80 kDa antigen molecules, next bind to 1,3-beta-glucan or chitin, probably via 1,6-beta-glucan, and, in an additional step, to chitin or 1,3-beta-glucan. This process results in the final molecular product of each antigen, and their distribution in the cell walls. PMID:11429475

  17. Linked in: immunologic membrane nanotube networks.

    Science.gov (United States)

    Zaccard, C R; Rinaldo, C R; Mailliard, R B

    2016-07-01

    Membrane nanotubes, also termed tunneling nanotubes, are F-actin-based structures that can form direct cytoplasmic connections and support rapid communication between distant cells. These nanoscale conduits have been observed in diverse cell types, including immune, neuronal, stromal, cancer, and stem cells. Until recently, little was known about the mechanisms involved in membrane nanotube development in myeloid origin APCs or how membrane nanotube networks support their ability to bridge innate and adaptive immunity. New research has provided insight into the modes of induction and regulation of the immune process of "reticulation" or the development of multicellular membrane nanotube networks in dendritic cells. Preprogramming by acute type 1 inflammatory mediators at their immature stage licenses mature type 1-polarized dendritic cells to reticulate upon subsequent interaction with CD40 ligand-expressing CD4(+) Th cells. Dendritic cell reticulation can support direct antigen transfer for amplification of specific T cell responses and can be positively or negatively regulated by signals from distinct Th cell subsets. Membrane nanotubes not only enhance the ability of immature dendritic cells to sense pathogens and rapidly mobilize nearby antigen-presenting cells in the peripheral tissues but also likely support communication of pathogen-related information from mature migratory dendritic cells to resident dendritic cells in lymph nodes. Therefore, the reticulation process facilitates a coordinated multicellular response for the efficient initiation of cell-mediated adaptive immune responses. Herein, we discuss studies focused on the molecular mechanisms of membrane nanotube formation, structure, and function in the context of immunity and how pathogens, such as HIV-1, may use dendritic cell reticulation to circumvent host defenses. PMID:26931578

  18. Nerve lesions following apical extrusion of non-setting calcium hydroxide: A systematic case review and report of two cases

    DEFF Research Database (Denmark)

    Olsen, Jesper Jared; Thorn, Jens Jørn; Korsgaard, Niels;

    2014-01-01

    We present two cases of apical extrusion of non-setting, calcium hydroxide paste which had been placed as an interappointment root canal dressing during routine endodontic treatment and which resulted in tissue necrosis of a large part of themandible, while. Surgical intervention consisted of...... effects and the use of an injectable system had most often been related to apical extrusion. Consequently, great care should be taken when applying the paste into the canal system. 2013 European Association for Cranio-Maxillo-Facial Surgery. Published by Elsevier Ltd. All rights reserved....

  19. Ultrastructure of the harmful unarmored dinoflagellate Cochlodinium polykrikoides (Dinophyceae) with reference to the apical groove and flagellar apparatus

    DEFF Research Database (Denmark)

    Iwataki, Mitsunori; Hansen, Gert; Moestrup, Øjvind;

    2010-01-01

    The external and internal ultrastructure of the harmful unarmored dinoflagellate Cochlodinium polykrikoides Margalef has been examined with special reference to the apical groove and three-dimensional structure of the flagellar apparatus. The apical groove is U-shaped and connected to the anterior...... eukaryotes but different from the Gymnodinium group with diagnostic nuclear chambers. The longitudinal and transverse basal bodies are separated by approximately 0.5-1.0 µm and interconnected directly by a striated basal body connective and indirectly by microtubular and fibrous structures. Characteristic...

  20. Electrophysiology of mycoplasma membranes.

    OpenAIRE

    Schummer, U.; Schiefer, H G

    1983-01-01

    The influence of transmembrane ion fluxes on mycoplasma membrane potentials was studied. Fluorescence intensity of potential-sensitive carbocyanine dyes was calibrated vs. electric membrane potential. Potassium and sodium ion diffusion potentials significantly contributed to mycoplasma membrane potential. Chloride ions were obviously freely permeable across mycoplasma membranes. Under growth conditions the mycoplasma membrane potential was estimated to be delta psi = - 80 mV.