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Sample records for apical membrane antigen

  1. Protein modeling of apical membrane antigen-1(AMA-1) of ...

    African Journals Online (AJOL)

    Apical membrane Antigen-1(AMA-1), an asexual blood stage antigen of Plasmodium cynomolgi, is an important candidate for testing as a component of malarial vaccine. The degree of conservation of. AMA-1 sequences implies a conserved function for this molecule across different species of Plasmodium. Since the AMA-1 ...

  2. ION-EXCHANGE IMMUNOAFFINITY PURIFICATION OF A RECOMBINANT BACULOVIRUS PLASMODIUM-FALCIPARUM APICAL MEMBRANE ANTIGEN, PF83/AMA-1

    NARCIS (Netherlands)

    NARUM, DL; WELLING, GW; THOMAS, AW

    1993-01-01

    A two-step purification regime has been developed for a quantitatively minor, putatively transmembrane, M(r) 83 000, apical membrane blood stage vaccine candidate antigen of Plasmodium falciparum (PF83/AMA-1), that has been expressed as a full-length baculovirus recombinant protein, PF83-FG8-1. The

  3. Identification of a Highly Antigenic Linear B Cell Epitope within Plasmodium vivax Apical Membrane Antigen 1 (AMA-1)

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    Bueno, Lilian Lacerda; Lobo, Francisco Pereira; Morais, Cristiane Guimarães; Mourão, Luíza Carvalho; de Ávila, Ricardo Andrez Machado; Soares, Irene Silva; Fontes, Cor Jesus; Lacerda, Marcus Vinícius; Olórtegui, Carlos Chavez; Bartholomeu, Daniella Castanheira; Fujiwara, Ricardo Toshio; Braga, Érika Martins

    2011-01-01

    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. PMID:21713006

  4. Molecular cloning, characterization and antigenicity of Babesia sp. BQ1 (Lintan) (Babesia cf. motasi) apical membrane antigen-1 (AMA-1).

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    Niu, Qingli; Liu, Zhijie; Yang, Jifei; Guan, Guiquan; Pan, Yuping; Luo, Jianxun; Yin, Hong

    2017-04-01

    Apical membrane antigen-1 (AMA-1) has been described as a potential vaccine candidate in apicomplexan parasites. Here we characterize the ama-1 gene. The full-length ama-1 gene of Babesia sp. BQ1 (Lintan) (BLTAMA-1) is 1785 bp, which contains an open reading frame (ORF) encoding a 65-kDa protein of 594 amino acid residues; by definition, the 5' UTR precedes the first methionine of the ORF. Phylogenetic analysis based on AMA-1 amino acid sequences clearly separated Piroplasmida from other Apicomplexa parasites. The Babesia sp. BQ1 (Lintan) AMA-1 sequence is most closely associated with that of B. ovata and B. bigemina, with high bootstrap value. A recombinant protein encoding a conserved region and containing ectodomains I and II of BLTAMA-1 was constructed. BLTrAMA-1-DI/DII proteins were tested for reactivity with sera from sheep infected by Babesia sp. BQ1 (Lintan). In Western-blot analysis, native Babesia sp. BQ1 (Lintan) AMA-1 proteins were recognized by antibodies raised in rabbits against BLTrAMA-1 in vitro. The results of this study are discussed in terms of gene characterization, taxonomy and antigenicity.

  5. 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.

  6. Population genetic structure and natural selection of apical membrane antigen-1 in Plasmodium vivax Korean isolates.

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    Kang, Jung-Mi; Lee, Jinyoung; Cho, Pyo-Yun; Moon, Sung-Ung; Ju, Hye-Lim; Ahn, Seong Kyu; Sohn, Woon-Mok; Lee, Hyeong-Woo; Kim, Tong-Soo; Na, Byoung-Kuk

    2015-11-16

    Plasmodium vivax apical membrane antigen-1 (PvAMA-1) is a leading candidate antigen for blood stage malaria vaccine. However, antigenic variation is a major obstacle in the development of an effective vaccine based on this antigen. In this study, the genetic structure and the effect of natural selection of PvAMA-1 among Korean P. vivax isolates were analysed. Blood samples were collected from 66 Korean patients with vivax malaria. The entire PvAMA-1 gene was amplified by polymerase chain reaction and cloned into a TA cloning vector. The PvAMA-1 sequence of each isolate was sequenced and the polymorphic characteristics and effect of natural selection were analysed using the DNASTAR, MEGA4, and DnaSP programs. Thirty haplotypes of PvAMA-1, which were further classified into seven different clusters, were identified in the 66 Korean P. vivax isolates. Domain II was highly conserved among the sequences, but substantial nucleotide diversity was observed in domains I and III. The difference between the rates of non-synonymous and synonymous mutations suggested that the gene has evolved under natural selection. No strong evidence indicating balancing or positive selection on PvAMA-1 was identified. Recombination may also play a role in the resulting genetic diversity of PvAMA-1. This study is the first comprehensive analysis of nucleotide diversity across the entire PvAMA-1 gene using a single population sample from Korea. Korean PvAMA-1 had limited genetic diversity compared to PvAMA-1 in global isolates. The overall pattern of genetic polymorphism of Korean PvAMA-1 differed from other global isolates and novel amino acid changes were also identified in Korean PvAMA-1. Evidences for natural selection and recombination event were observed, which is likely to play an important role in generating genetic diversity across the PvAMA-1. These results provide useful information for the understanding the population structure of P. vivax circulating in Korea and have important

  7. Population genetic structure and natural selection of Plasmodium falciparum apical membrane antigen-1 in Myanmar isolates.

    Science.gov (United States)

    Kang, Jung-Mi; Lee, Jinyoung; Moe, Mya; Jun, Hojong; Lê, Hương Giang; Kim, Tae Im; Thái, Thị Lam; Sohn, Woon-Mok; Myint, Moe Kyaw; Lin, Khin; Shin, Ho-Joon; Kim, Tong-Soo; Na, Byoung-Kuk

    2018-02-07

    Plasmodium falciparum apical membrane antigen-1 (PfAMA-1) is one of leading blood stage malaria vaccine candidates. However, genetic variation and antigenic diversity identified in global PfAMA-1 are major hurdles in the development of an effective vaccine based on this antigen. In this study, genetic structure and the effect of natural selection of PfAMA-1 among Myanmar P. falciparum isolates were analysed. Blood samples were collected from 58 Myanmar patients with falciparum malaria. Full-length PfAMA-1 gene was amplified by polymerase chain reaction and cloned into a TA cloning vector. PfAMA-1 sequence of each isolate was sequenced. Polymorphic characteristics and effect of natural selection were analysed with using DNASTAR, MEGA4, and DnaSP programs. Polymorphic nature and natural selection in 459 global PfAMA-1 were also analysed. Thirty-seven different haplotypes of PfAMA-1 were identified in 58 Myanmar P. falciparum isolates. Most amino acid changes identified in Myanmar PfAMA-1 were found in domains I and III. Overall patterns of amino acid changes in Myanmar PfAMA-1 were similar to those in global PfAMA-1. However, frequencies of amino acid changes differed by country. Novel amino acid changes in Myanmar PfAMA-1 were also identified. Evidences for natural selection and recombination event were observed in global PfAMA-1. Among 51 commonly identified amino acid changes in global PfAMA-1 sequences, 43 were found in predicted RBC-binding sites, B-cell epitopes, or IUR regions. Myanmar PfAMA-1 showed similar patterns of nucleotide diversity and amino acid polymorphisms compared to those of global PfAMA-1. Balancing natural selection and intragenic recombination across PfAMA-1 are likely to play major roles in generating genetic diversity in global PfAMA-1. Most common amino acid changes in global PfAMA-1 were located in predicted B-cell epitopes where high levels of nucleotide diversity and balancing natural selection were found. These results highlight the

  8. 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

  9. 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

  10. Temporal genetic changes in Plasmodium vivax apical membrane antigen 1 over 19 years of transmission in southern Mexico.

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    Flores-Alanis, Alejandro; González-Cerón, Lilia; Santillán, Frida; Ximenez, Cecilia; Sandoval, Marco A; Cerritos, René

    2017-05-02

    Mexico advanced to the pre-elimination phase in 2009 due to a significant reduction in malaria cases, and since 2000, Plasmodium vivax is the only species transmitted. During the last two decades, malaria transmission has been mostly local and isolated to a few regions. It is important to gain further insights into the impact of control measures on the parasite population structure. Hence, the aim of the current study was to determine detailed changes in P. vivax genetic diversity and population structure based on analysing the gene that encodes the apical membrane antigen 1 (pvama1). This analysis covered from control to pre-elimination (1993-2011) in a hypo-endemic region in southern Mexico. The 213 pvama1 I-II sequences presently analysed were grouped into six periods of three years each. They showed low genetic diversity, with 15 haplotypes resolved. Among the DNA sequences, there was a gradual decrease in genetic diversity, the number of mixed genotype infections and the intensity of positive selection, in agreement with the parallel decline in malaria cases. At the same time, linkage disequilibrium (R 2 ) increased. The three-dimensional haplotype network revealed that pvama1 I-II haplotypes were separated by 1-11 mutational steps, and between one another by 0-3 unsampled haplotypes. In the temporal network, seven haplotypes were detected in at least two of the six-time layers, and only four distinct haplotypes were evidenced in the pre-elimination phase. Structure analysis indicated that three subpopulations fluctuated over time. Only 8.5% of the samples had mixed ancestry. In the pre-elimination phase, subpopulation P1 was drastically reduced, and the admixture was absent. The results suggest that P. vivax in southern Mexico evolved based on local adaptation into three "pseudoclonal" subpopulations that diversified at the regional level and persisted over time, although with varying frequency. Control measures and climate events influenced the number of

  11. 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

  12. Phase 1/2a study of the malaria vaccine candidate apical membrane antigen-1 (AMA-l) administered in adjuvant system AS01B or AS02A

    NARCIS (Netherlands)

    M.D. Spring (Michele Donna); J.F. Cummings (James); C.F. Ockenhouse (Christian); S. Dutta (Shantanu); R. Reidler (Randall); E. Angov (Evelina); E. Bergmann-Leitner (Elke); V.A. Stewart (Ann); S. Bittner (Stacey); L. Juompan (Laure); M.G. Kortepeter (Mark); R. Nielsen (Robin); U. Krzych (Urszula); E. Tierney (Ev); L.A. Ware (Lisa); M. Dowler (Megan); C.C. Hermsen (Cornelus); R.W. Sauerwein (Robert); S.J. de Vlas (Sake); O. Ofori-Anyinam (Opokua); D.E. Lanar (David); J.L. Williams (Jack); K.E. Kester (Kent); K. Tucker (Kathryn); M. Shi (Meng); E. Malkin (Elissa); C. Long (Carole); C.L. Diggs (Carter); L. Soisson (Lorraine Amory); M.C. Dubois; W.R. Ballou (Ripley); J. Cohen (Joe); D.G. Heppner (Gray)

    2009-01-01

    textabstractBackground: This Phase 1/2a study evaluated the safety, immunogenicity, and efficacy of an experimental malaria vaccine comprised of the recombinant Plasmodium falciparum protein apical membrane antigen-1 (AMA-1) representing the 3D7 allele formulated with either the AS01B or AS02A

  13. 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

    2010-06-01

    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.

  14. Phase 1/2a study of the malaria vaccine candidate apical membrane antigen-1 (AMA-1 administered in adjuvant system AS01B or AS02A.

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    Michele D Spring

    Full Text Available This Phase 1/2a study evaluated the safety, immunogenicity, and efficacy of an experimental malaria vaccine comprised of the recombinant Plasmodium falciparum protein apical membrane antigen-1 (AMA-1 representing the 3D7 allele formulated with either the AS01B or AS02A Adjuvant Systems.After a preliminary safety evaluation of low dose AMA-1/AS01B (10 microg/0.5 mL in 5 adults, 30 malaria-naïve adults were randomly allocated to receive full dose (50 microg/0.5 mL of AMA-1/AS01B (n = 15 or AMA-1/AS02A (n = 15, followed by a malaria challenge. All vaccinations were administered intramuscularly on a 0-, 1-, 2-month schedule. All volunteers experienced transient injection site erythema, swelling and pain. Two weeks post-third vaccination, anti-AMA-1 Geometric Mean Antibody Concentrations (GMCs with 95% Confidence Intervals (CIs were high: low dose AMA-1/AS01B 196 microg/mL (103-371 microg/mL, full dose AMA-1/AS01B 279 microg/mL (210-369 microg/mL and full dose AMA-1/AS02A 216 microg/mL (169-276 microg/mL with no significant difference among the 3 groups. The three vaccine formulations elicited equivalent functional antibody responses, as measured by growth inhibition assay (GIA, against homologous but not against heterologous (FVO parasites as well as demonstrable interferon-gamma (IFN-gamma responses. To assess efficacy, volunteers were challenged with P. falciparum-infected mosquitoes, and all became parasitemic, with no significant difference in the prepatent period by either light microscopy or quantitative polymerase chain reaction (qPCR. However, a small but significant reduction of parasitemia in the AMA-1/AS02A group was seen with a statistical model employing qPCR measurements.All three vaccine formulations were found to be safe and highly immunogenic. These immune responses did not translate into significant vaccine efficacy in malaria-naïve adults employing a primary sporozoite challenge model, but encouragingly, estimation of parasite

  15. Non-apical membrane antigen 1 (AMA1 IgGs from Malian children interfere with functional activity of AMA1 IgGs as judged by growth inhibition assay.

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    Kazutoyo Miura

    Full Text Available BACKGROUND: Apical membrane antigen 1 (AMA1 is one of the best-studied blood-stage malaria vaccine candidates. When an AMA1 vaccine was tested in a malaria naïve population, it induced functionally active antibodies judged by Growth Inhibition Assay (GIA. However, the same vaccine failed to induce higher growth-inhibitory activity in adults living in a malaria endemic area. Vaccination did induce functionally active antibodies in malaria-exposed children with less than 20% inhibition in GIA at baseline, but not in children with more than that level of baseline inhibition. METHODS: Total IgGs were purified from plasmas collected from the pediatric trial before and after immunization and pools of total IgGs were made. Another set of total IgGs was purified from U.S. adults immunized with AMA1 (US-total IgG. From these total IgGs, AMA1-specific and non-AMA1 IgGs were affinity purified and the functional activity of these IgGs was evaluated by GIA. Competition ELISA was performed with the U.S.-total IgG and non-AMA1 IgGs from malaria-exposed children. RESULTS: AMA1-specific IgGs from malaria-exposed children and U.S. vaccinees showed similar growth-inhibitory activity at the same concentrations. When mixed with U.S.-total IgG, non-AMA1 IgGs from children showed an interference effect in GIA. Interestingly, the interference effect was higher with non-AMA1 IgGs from higher titer pools. The non-AMA1 IgGs did not compete with anti-AMA1 antibody in U.S.-total IgG in the competition ELISA. CONCLUSION: Children living in a malaria endemic area have a fraction of IgGs that interferes with the biological activity of anti-AMA1 antibody as judged by GIA. While the mechanism of interference is not resolved in this study, these results suggest it is not caused by direct competition between non-AMA1 IgG and AMA1 protein. This study indicates that anti-malaria IgGs induced by natural exposure may interfere with the biological effect of antibody induced by an AMA1

  16. Determination of apical membrane polarity in mammary epithelial cell cultures: The role of cell-cell, cell-substratum, and membrane-cytoskeleton interactions

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    Parry, G.; Beck, J.C.; Moss, L.; Bartley, J. (Lawrence Berkeley Lab., CA (United States)); Ojakian, G.K. (State Univ. of New York, Brooklyn (United States))

    1990-06-01

    The membrane glycoprotein, PAS-O, is a major differentiation antigen on mammary epithelial cells and is located exclusively in the apical domain of the plasma membrane. The authors have used 734B cultured human mammary carcinoma cells as a model system to study the role of tight junctions, cell-substratum contacts, and submembranous cytoskeletal elements in restricting PAS-O to the apical membrane. Immunofluorescence and immunoelectronmicroscopy experiments demonstrated that while tight junctions demarcate PAS-O distribution in confluent cultures, apical polarity could be established at low culture densities when cells could not form tight junctions with neighboring cells. They suggest, then, that interactions between vitronectin and its receptor, are responsible for establishment of membrane domains in the absence of tight junctions. The role of cytoskeletal elements in restricting PAS-O distribution was examined by treating cultures with cytochalasin D, colchicine, or acrylamide. Cytochalasin D led to a redistribution of PAS0O while colchicine and acrylamide did not. They hypothesize that PAS-O is restricted to the apical membrane by interactions with a microfilament network and that the cytoskeletal organization is dependent upon cell-cell and cell-substratum interactions.

  17. Outer membrane protein antigens of Moraxella bovis.

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    Ostle, A G; Rosenbusch, R F

    1986-07-01

    Outer membranes were isolated from bovine isolates and type strains of Moraxella bovis, M phenylpyruvica, M lacunata, and M ovis by sodium N lauroyl sarcosinate extraction and differential centrifugation. Analysis of outer membranes from these organisms by sodium dodecyl sulfate-polyacrylamide-gel electrophoresis revealed that all M bovis isolates shared a common polypeptide pattern that was readily distinguishable from other Moraxella spp. Nine major outer membrane protein bands were identified by sodium dodecyl sulfate-polyacrylamide-gel electrophoresis analysis of M bovis. Immunoblotting of protein antigens of M bovis revealed several outer membrane proteins that seemed to be common antigens of all M bovis isolates.

  18. Basolateral cholesterol depletion alters Aquaporin-2 post-translational modifications and disrupts apical plasma membrane targeting.

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    Moeller, Hanne B; Fuglsang, Cecilia Hvitfeldt; Pedersen, Cecilie Nøhr; Fenton, Robert A

    2018-01-01

    Apical plasma membrane accumulation of the water channel Aquaporin-2 (AQP2) in kidney collecting duct principal cells is critical for body water homeostasis. Posttranslational modification (PTM) of AQP2 is important for regulating AQP2 trafficking. The aim of this study was to determine the role of cholesterol in regulation of AQP2 PTM and in apical plasma membrane targeting of AQP2. Cholesterol depletion from the basolateral plasma membrane of a collecting duct cell line (mpkCCD14) using methyl-beta-cyclodextrin (MBCD) increased AQP2 ubiquitylation. Forskolin, cAMP or dDAVP-mediated AQP2 phosphorylation at Ser269 (pS269-AQP2) was prevented by cholesterol depletion from the basolateral membrane. None of these effects on pS269-AQP2 were observed when cholesterol was depleted from the apical side of cells, or when MBCD was applied subsequent to dDAVP stimulation. Basolateral, but not apical, MBCD application prevented cAMP-induced apical plasma membrane accumulation of AQP2. These studies indicate that manipulation of the cholesterol content of the basolateral plasma membrane interferes with AQP2 PTM and subsequently regulated apical plasma membrane targeting of AQP2. Copyright © 2017 Elsevier Inc. All rights reserved.

  19. 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.

  20. Trafficking Ion Transporters to the Apical Membrane of Polarized Intestinal Enterocytes.

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    Engevik, Amy Christine; Goldenring, James R

    2018-01-02

    Epithelial cells lining the gastrointestinal tract require distinct apical and basolateral domains to function properly. Trafficking and insertion of enzymes and transporters into the apical brush border of intestinal epithelial cells is essential for effective digestion and absorption of nutrients. Specific critical ion transporters are delivered to the apical brush border to facilitate fluid and electrolyte uptake. Maintenance of these apical transporters requires both targeted delivery and regulated membrane recycling. Examination of altered apical trafficking in patients with Microvillus Inclusion disease caused by inactivating mutations in MYO5B has led to insights into the regulation of apical trafficking by elements of the apical recycling system. Modeling of MYO5B loss in cell culture and animal models has led to recognition of Rab11a and Rab8a as critical regulators of apical brush border function. All of these studies show the importance of apical membrane trafficking dynamics in maintenance of polarized epithelial cell function. Copyright © 2018 Cold Spring Harbor Laboratory Press; all rights reserved.

  1. Par3 integrates Tiam1 and phosphatidylinositol 3-kinase signaling to change apical membrane identity.

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    Ruch, Travis R; Bryant, David M; Mostov, Keith E; Engel, Joanne N

    2017-01-15

    Pathogens can alter epithelial polarity by recruiting polarity proteins to the apical membrane, but how a change in protein localization is linked to polarity disruption is not clear. In this study, we used chemically induced dimerization to rapidly relocalize proteins from the cytosol to the apical surface. We demonstrate that forced apical localization of Par3, which is normally restricted to tight junctions, is sufficient to alter apical membrane identity through its interactions with phosphatidylinositol 3-kinase (PI3K) and the Rac1 guanine nucleotide exchange factor Tiam1. We further show that PI3K activity is required upstream of Rac1, and that simultaneously targeting PI3K and Tiam1 to the apical membrane has a synergistic effect on membrane remodeling. Thus, Par3 coordinates the action of PI3K and Tiam1 to define membrane identity, revealing a signaling mechanism that can be exploited by human mucosal pathogens. © 2017 Ruch et al. This article is distributed by The American Society for Cell Biology under license from the author(s). Two months after publication it is available to the public under an Attribution–Noncommercial–Share Alike 3.0 Unported Creative Commons License (http://creativecommons.org/licenses/by-nc-sa/3.0).

  2. Lats kinase is involved in the intestinal apical membrane integrity in the nematode Caenorhabditis elegans.

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    Kang, Junsu; Shin, Donghoon; Yu, Jae-Ran; Lee, Junho

    2009-08-01

    The roles of Lats kinases in the regulation of cell proliferation and apoptosis have been well established. Here we report new roles for Lats kinase in the integrity of the apical membrane structure. WTS-1, the C. elegans Lats homolog, localized primarily to the subapical region in the intestine. A loss-of-function mutation in wts-1 resulted in an early larval arrest and defects in the structure of the intestinal lumen. An electron microscopy study of terminally arrested wts-1 mutant animals revealed numerous microvilli-containing lumen-like structures within the intestinal cells. The wts-1 phenotype was not caused by cell proliferation or apoptosis defects. Instead, we found that the wts-1 mutant animals exhibited gradual mislocalization of apical actin and apical junction proteins, suggesting that wts-1 normally suppresses the formation of extra apical membrane structures. Heat-shock-driven pulse-chase expression experiments showed that WTS-1 regulates the localization of newly synthesized apical actins. RNAi of the exocyst complex genes suppressed the mislocalization phenotype of wts-1 mutation. Collectively, the data presented here suggest that Lats kinase plays important roles in the integrity of the apical membrane structure of intestinal cells.

  3. Protein modeling of apical membrane antigen-1(AMA-1) of ...

    African Journals Online (AJOL)

    SERVER

    2007-11-19

    Nov 19, 2007 ... available for the vast majority of the protein sequences. At this juncture, prediction methods have gained much. *Corresponding author. E-mail: drdmmamatha@gmail.com interest (Schwede et al. 2003). Genome sequencing projects continuously detect new protein sequences, this provides new information ...

  4. Src- and Fyn-dependent apical membrane trafficking events control endothelial lumen formation during vascular tube morphogenesis.

    Science.gov (United States)

    Kim, Dae Joong; Norden, Pieter R; Salvador, Jocelynda; Barry, David M; Bowers, Stephanie L K; Cleaver, Ondine; Davis, George E

    2017-01-01

    Here we examine the question of how endothelial cells (ECs) develop their apical membrane surface domain during lumen and tube formation. We demonstrate marked apical membrane targeting of activated Src kinases to this apical domain during early and late stages of this process. Immunostaining for phosphotyrosine or phospho-Src reveals apical membrane staining in intracellular vacuoles initially. This is then followed by vacuole to vacuole fusion events to generate an apical luminal membrane, which is similarly decorated with activated phospho-Src kinases. Functional blockade of Src kinases completely blocks EC lumen and tube formation, whether this occurs during vasculogenic tube assembly or angiogenic sprouting events. Multiple Src kinases participate in this apical membrane formation process and siRNA suppression of Src, Fyn and Yes, but not Lyn, blocks EC lumen formation. We also demonstrate strong apical targeting of Src-GFP and Fyn-GFP fusion proteins and increasing their expression enhances lumen formation. Finally, we show that Src- and Fyn-associated vacuoles track and fuse along a subapically polarized microtubule cytoskeleton, which is highly acetylated. These vacuoles generate the apical luminal membrane in a stereotypically polarized, perinuclear position. Overall, our study identifies a critical role for Src kinases in creating and decorating the EC apical membrane surface during early and late stages of lumen and tube formation, a central event in the molecular control of vascular morphogenesis.

  5. Src- and Fyn-dependent apical membrane trafficking events control endothelial lumen formation during vascular tube morphogenesis.

    Directory of Open Access Journals (Sweden)

    Dae Joong Kim

    Full Text Available Here we examine the question of how endothelial cells (ECs develop their apical membrane surface domain during lumen and tube formation. We demonstrate marked apical membrane targeting of activated Src kinases to this apical domain during early and late stages of this process. Immunostaining for phosphotyrosine or phospho-Src reveals apical membrane staining in intracellular vacuoles initially. This is then followed by vacuole to vacuole fusion events to generate an apical luminal membrane, which is similarly decorated with activated phospho-Src kinases. Functional blockade of Src kinases completely blocks EC lumen and tube formation, whether this occurs during vasculogenic tube assembly or angiogenic sprouting events. Multiple Src kinases participate in this apical membrane formation process and siRNA suppression of Src, Fyn and Yes, but not Lyn, blocks EC lumen formation. We also demonstrate strong apical targeting of Src-GFP and Fyn-GFP fusion proteins and increasing their expression enhances lumen formation. Finally, we show that Src- and Fyn-associated vacuoles track and fuse along a subapically polarized microtubule cytoskeleton, which is highly acetylated. These vacuoles generate the apical luminal membrane in a stereotypically polarized, perinuclear position. Overall, our study identifies a critical role for Src kinases in creating and decorating the EC apical membrane surface during early and late stages of lumen and tube formation, a central event in the molecular control of vascular morphogenesis.

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

    DEFF Research Database (Denmark)

    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......, the rates of the Golgi-associated complex glycosylation and association with rafts of newly synthesized aminopeptidase N were reduced, and less of the enzyme had reached the brush border membrane after 2 h of labeling. In contrast, the basolateral Na(+)/K(+)-ATPase was neither missorted nor raft...

  7. Polarized Human Retinal Pigment Epithelium Exhibits Distinct Surface Proteome on Apical and Basal Plasma Membranes.

    Science.gov (United States)

    Khristov, Vladimir; Wan, Qin; Sharma, Ruchi; Lotfi, Mostafa; Maminishkis, Arvydas; Bharti, Kapil

    2018-01-01

    Surface proteins localized on the apical and basal plasma membranes are required for a cell to sense its environment and relay changes in ionic, cytokine, chemokine, and hormone levels to the inside of the cell. In a polarized cell, surface proteins are differentially localized on the apical or the basolateral sides of the cell. The retinal pigment epithelium (RPE) is an example of a polarized cell that performs a variety of functions that are dependent on its polarized state including trafficking of ions, fluid, and metabolites across the RPE monolayer. These functions are absolutely crucial for maintaining the health and integrity of adjacent photoreceptors, the photosensitive cells of the retina. Here we present a series of approaches to identify and validate the polarization state of cultured primary human RPE cells using immunostaining for RPE apical/basolateral markers, polarized cytokine secretion, electrophysiology, fluid transport, phagocytosis, and identification of plasma membrane proteins through cell surface capturing technology. These approaches are currently being used to validate the polarized state and the epithelial phenotype of human induced pluripotent stem (iPS) cell derived RPE cells. This work provides the basis for developing an autologous cell therapy for age-related macular degeneration using patient specific iPS cell derived RPE.

  8. Oncostatin M-stimulated apical plasma membrane biogenesis requires p27(Kip1)-regulated cell cycle dynamics

    NARCIS (Netherlands)

    Van IJzendoorn, Sven C D; Théard, Delphine; Van Der Wouden, Johanna M; Visser, Willy; Wojtal, Kacper A; Hoekstra, Dick

    Oncostatin M regulates membrane traffic and stimulates apicalization of the cell surface in hepatoma cells in a protein kinase A-dependent manner. Here, we show that oncostatin M enhances the expression of the cyclin-dependent kinase (cdk)2 inhibitor p27(Kip1), which inhibits G(1)-S-phase

  9. Prostate-specific membrane antigen and its truncated form PSM'

    Czech Academy of Sciences Publication Activity Database

    Mlčochová, Petra; Bařinka, Cyril; Tykvart, Jan; Šácha, Pavel; Konvalinka, Jan

    2009-01-01

    Roč. 69, č. 5 (2009), s. 471-479 ISSN 0270-4137 R&D Projects: GA MŠk 1M0508 Institutional research plan: CEZ:AV0Z40550506 Keywords : prostate specific membrane antigen * glutamate carboxypeptidase II * prostate cancer Subject RIV: CE - Biochemistry Impact factor: 3.081, year: 2009

  10. Peptidases Compartmentalized to the Ascaris suum Intestinal Lumen and Apical Intestinal Membrane

    Science.gov (United States)

    Rosa, Bruce A.

    2015-01-01

    The nematode intestine is a tissue of interest for developing new methods of therapy and control of parasitic nematodes. However, biological details of intestinal cell functions remain obscure, as do the proteins and molecular functions located on the apical intestinal membrane (AIM), and within the intestinal lumen (IL) of nematodes. Accordingly, methods were developed to gain a comprehensive identification of peptidases that function in the intestinal tract of adult female Ascaris suum. Peptidase activity was detected in multiple fractions of the A. suum intestine under pH conditions ranging from 5.0 to 8.0. Peptidase class inhibitors were used to characterize these activities. The fractions included whole lysates, membrane enriched fractions, and physiological- and 4 molar urea-perfusates of the intestinal lumen. Concanavalin A (ConA) was confirmed to bind to the AIM, and intestinal proteins affinity isolated on ConA-beads were compared to proteins from membrane and perfusate fractions by mass spectrometry. Twenty-nine predicted peptidases were identified including aspartic, cysteine, and serine peptidases, and an unexpectedly high number (16) of metallopeptidases. Many of these proteins co-localized to multiple fractions, providing independent support for localization to specific intestinal compartments, including the IL and AIM. This unique perfusion model produced the most comprehensive view of likely digestive peptidases that function in these intestinal compartments of A. suum, or any nematode. This model offers a means to directly determine functions of these proteins in the A. suum intestine and, more generally, deduce the wide array functions that exist in these cellular compartments of the nematode intestine. PMID:25569475

  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. Aldosterone induction of electrogenic sodium transport in the apical membrane vesicles of rat distal colon

    International Nuclear Information System (INIS)

    Rajendran, V.M.; Kashgarian, M.; Binder, H.J.

    1989-01-01

    Na-H exchange is present in apical membrane vesicles (AMV) isolated from distal colon of normal rats. Because in intact tissue aldosterone both induces amiloride-sensitive electrogenic sodium transport and inhibits electroneutral sodium absorption, these studies with AMV were designed to establish the effect of aldosterone on sodium transport. An outward-directed proton gradient stimulated 22Na uptake in AMV isolated from distal colon of normal and dietary sodium depleted (with elevated aldosterone levels) experimental rats. Unlike normal AMV, proton gradient-dependent 22Na uptake in experimental AMV was inhibited when uptake was measured under voltage-clamped conditions. 10 microM amiloride inhibited the initial rate of proton gradient-dependent 22Na uptake in AMV of normal and experimental rats by 30 and 75%, respectively. In contrast, 1 mM amiloride produced comparable inhibition (90 and 80%) of 22Na uptake in normal and experimental AMV. Intravesicular-negative potential stimulated 22Na uptake in experimental but not in normal AMV. This increase was inhibited by 90% by 10 microM amiloride. An analogue of amiloride, 5-(N-ethylisopropyl) amiloride (1 microM), a potent inhibitor of electroneutral Na-H exchange in AMV of normal rat distal colon, did not alter potassium diffusion potential-dependent 22Na uptake. Increasing sodium concentration saturated proton gradient-dependent 22Na uptake in normal AMV. However, in experimental AMV, 22Na uptake stimulated by both proton gradient and potassium diffusion potential did not saturate as a function of increasing sodium concentration. We conclude from these results that an electrically sensitive conductive channel, not electroneutral Na-H exchange, mediates 22Na uptake in AMV isolated from the distal colon of aldosterone rats

  13. Na+/HCO3- Cotransporter NBCn2 Mediates HCO3- Reclamation in the Apical Membrane of Renal Proximal Tubules.

    Science.gov (United States)

    Guo, Yi-Min; Liu, Ying; Liu, Mei; Wang, Jin-Lin; Xie, Zhang-Dong; Chen, Kang-Jing; Wang, Deng-Ke; Occhipinti, Rossana; Boron, Walter F; Chen, Li-Ming

    2017-08-01

    The kidney maintains systemic acid-base balance by reclaiming from the renal tubule lumen virtually all HCO 3 - filtered in glomeruli and by secreting additional H + to titrate luminal buffers. For proximal tubules, which are responsible for about 80% of this activity, it is believed that HCO 3 - reclamation depends solely on H + secretion, mediated by the apical Na + /H + exchanger NHE 3 and the vacuolar proton pump. However, NHE3 and the proton pump cannot account for all HCO 3 - reclamation. Here, we investigated the potential contribution of two variants of the electroneutral Na + /HCO 3 - cotransporter NBCn2, the amino termini of which start with the amino acids MCDL (MCDL-NBCn2) and MEIK (MEIK-NBCn2). Western blot analysis and immunocytochemistry revealed that MEIK-NBCn2 predominantly localizes at the basolateral membrane of medullary thick ascending limbs in the rat kidney, whereas MCDL-NBCn2 localizes at the apical membrane of proximal tubules. Notably, NH 4 Cl-induced systemic metabolic acidosis or hypokalemic alkalosis downregulated the abundance of MCDL-NBCn2 and reciprocally upregulated NHE 3 Conversely, NaHCO 3 -induced metabolic alkalosis upregulated MCDL-NBCn2 and reciprocally downregulated NHE 3 We propose that the apical membrane of the proximal tubules has two distinct strategies for HCO 3 - reclamation: the conventional indirect pathway, in which NHE 3 and the proton pump secrete H + to titrate luminal HCO 3 - , and the novel direct pathway, in which NBCn2 removes HCO 3 - from the lumen. The reciprocal regulation of NBCn2 and NHE 3 under different physiologic conditions is consistent with our mathematical simulations, which suggest that HCO 3 - uptake and H + secretion have reciprocal efficiencies for HCO 3 - reclamation versus titration of luminal buffers. Copyright © 2017 by the American Society of Nephrology.

  14. New saliva secretion model based on the expression of Na+-K+ pump and K+ channels in the apical membrane of parotid acinar cells.

    Science.gov (United States)

    Almássy, János; Siguenza, Elias; Skaliczki, Marianna; Matesz, Klara; Sneyd, James; Yule, David I; Nánási, Péter P

    2018-04-01

    The plasma membrane of parotid acinar cells is functionally divided into apical and basolateral regions. According to the current model, fluid secretion is driven by transepithelial ion gradient, which facilitates water movement by osmosis into the acinar lumen from the interstitium. The osmotic gradient is created by the apical Cl - efflux and the subsequent paracellular Na + transport. In this model, the Na + -K + pump is located exclusively in the basolateral membrane and has essential role in salivary secretion, since the driving force for Cl - transport via basolateral Na + -K + -2Cl - cotransport is generated by the Na + -K + pump. In addition, the continuous electrochemical gradient for Cl - flow during acinar cell stimulation is maintained by the basolateral K + efflux. However, using a combination of single-cell electrophysiology and Ca 2+ -imaging, we demonstrate that photolysis of Ca 2+ close to the apical membrane of parotid acinar cells triggered significant K + current, indicating that a substantial amount of K + is secreted into the lumen during stimulation. Nevertheless, the K + content of the primary saliva is relatively low, suggesting that K + might be reabsorbed through the apical membrane. Therefore, we investigated the localization of Na + -K + pumps in acinar cells. We show that the pumps appear evenly distributed throughout the whole plasma membrane, including the apical pole of the cell. Based on these results, a new mathematical model of salivary fluid secretion is presented, where the pump reabsorbs K + from and secretes Na + to the lumen, which can partially supplement the paracellular Na + pathway.

  15. Cloning and characterization of canine prostate-specific membrane antigen.

    Science.gov (United States)

    Schmidt, Sonja; Fracasso, Giulio; Colombatti, Marco; Naim, Hassan Y

    2013-05-01

    Prostate-specific membrane antigen (PSMA) is a promising biomarker in the diagnosis of prostate cancer and a potential target for antibody-based therapeutic strategies. We isolated the canine PSMA cDNA and investigated the cellular and biochemical characteristics of the recombinant protein as a potential target for animal preclinical studies of antibody based-therapies. Canine PSMA cDNA was isolated by PCR, cloned into expression vectors and transfected into COS-1 and MDCK cells. The biosynthesis and glycosylation of the recombinant protein were investigated in pulse-chase experiments, the cellular localization by confocal laser microscopy, the mode of association of PSMA with the membrane with solubilization in different detergents and its quaternary structure in sucrose-density gradients. Canine PSMA shows 91% amino acid homology to human PSMA, whereby the major difference is a longer cytoplasmic tail of canine PSMA compared to its human counterpart. Canine PSMA is trafficked efficiently along the secretory pathway, undergoes homodimerization when it acquires complex glycosylated mature form. It associates with detergent-resistant membranes, which act as platforms along its intracellular trafficking. Confocal analysis revealed canine PSMA at the cell surface, Golgi, and the endoplasmic reticulum. A similar distribution is revealed for human PSMA, yet with reduced cell surface levels. The cloning, expression, biosynthesis, processing and localization of canine PSMA in mammalian cells is described. We demonstrate that canine PSMA reveals similar characteristics to human PSMA rendering this protein useful as a translational model for investigations of prostate cancer as well as a suitable antigen for targeted therapy studies in dogs. Copyright © 2013 Wiley Periodicals, Inc.

  16. Radiolabeled prostate-specific membrane antigen small-molecule inhibitors.

    Science.gov (United States)

    Will, Leon; Sonni, Ida; Kopka, Klaus; Kratochwil, Clemens; Giesel, Frederik L; Haberkorn, Uwe

    2017-06-01

    Prostate cancer (PC) is one of the most common malignancies worldwide. Prostate-specific membrane antigen (PSMA) has been found to be expressed in most PCs and represents an ideal target for diagnostic and therapeutic purposes. Numerous PSMA tracers have been recently developed. This review aims to provide an overview on the clinical influence of PSMA tracers in primary staging, biochemical recurrence (BCR) of PC and advanced, metastatic PC. Additionally, the use of PSMA tracers in systemic radioligand therapy (RLT) of metastatic castration-resistant prostate cancer (mCRPC), as well as non-prostatic specific uptake of PSMA tracers and the use of PSMA imaging to manage therapy have been described. A computerized search of the literature (PubMed) was conducted in order to find evidence on the role of PSMA tracers in the diagnosis and therapy of PC. PSMA positron-emission tomography/computed tomography (PET/CT) outperforms conventional imaging in the settings of primary PC, BCR and advanced PC. Especially in BCR of PC, PSMA PET/CT shows clinical value with significantly higher detection rates than standard modalities. The use of PSMA PET/CT resulted in a change of the therapeutic management in up to half of the cases. Regarding RLT, smaller studies were able to show positive clinical effects of 177Lu-labeled PSMA tracers without the occurrence of severe side effects. The currently available data clearly shows that PSMA targeting has a clinical impact on the diagnosis of PC, and that RLT using radiolabeled PSMA tracers has high potentiality in the settings of resistance to conventional therapeutic approaches.

  17. Dynamics of the membrane-cytoskeleton interface in MHC class II-restricted antigen presentation.

    Science.gov (United States)

    Bretou, Marine; Kumari, Anita; Malbec, Odile; Moreau, Hélène D; Obino, Dorian; Pierobon, Paolo; Randrian, Violaine; Sáez, Pablo J; Lennon-Duménil, Ana-Maria

    2016-07-01

    Antigen presentation refers to the ability of cells to show MHC-associated determinants to T lymphocytes, leading to their activation. MHC class II molecules mainly present peptide-derived antigens that are internalized by endocytosis in antigen-presenting cells (APCs). Here, we describe how the interface between cellular membranes and the cytoskeleton regulates the various steps that lead to the presentation of exogenous antigens on MHC class II molecules in the two main types of APCs: dendritic cells (DCs) and B lymphocytes. This includes antigen uptake, processing, APC migration, and APC-T cell interactions. We further discuss how the interaction between APC-specific molecules and cytoskeleton elements allows the coordination of antigen presentation and cell migration in time and space. © 2016 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

  18. Effects of radiation on the expression of antigens on the membranes of human adenocarcinoma cells

    International Nuclear Information System (INIS)

    Hareyama, Masato; Imai, Kozo; Oouchi, Atsushi; Shidou, Mitsuo; Takahashi, Hiroki; Koshiba, Hirohumi; Yachi, Akira; Morita, Kazuo

    1992-01-01

    We have investigated the effects of irradiation on the membranes of tumor cells. X-ray irradiation caused remarkable increases in the expression of tumor-associated antigens (YH206, CEA) and c-erbB-2 protein by flow cytometry, whereas IFN had no obvious effect on the expression of tumor-associated antigens and c-erbB-2 protein. On the other hand, the expression of MHC Class I and ICAM-1 antigen on the membrane by flow cytometry was enhanced by both irradiation and IFN. In addition, the irradiated cells, when analyzed using a CEA specific probe, showed remarkable increases in the CEA mRNA compared to IFN-treated cells. It is possible that enhancement of the expression of tumor-associated antigens and c-erbB-2 protein, together with the enhancement of that of MHC-Class I and ICAM-1, would help cytotoxic killer cells recognize the tumor cells. (author)

  19. The Whitish Inner Mantle of the Giant Clam, Tridacna squamosa, Expresses an Apical Plasma Membrane Ca2+-ATPase (PMCA Which Displays Light-Dependent Gene and Protein Expressions

    Directory of Open Access Journals (Sweden)

    Yuen K. Ip

    2017-10-01

    Full Text Available Giant clams live in symbiosis with extracellular zooxanthellae and display high rates of growth and shell formation (calcification in light. Light-enhanced calcification requires an increase in the supply of Ca2+ to, and simultaneously an augmented removal of H+ from, the extrapallial fluid where shell formation occurs. We have obtained the complete coding cDNA sequence of Plasma Membrane Ca2+-ATPase (PMCA from the thin and whitish inner mantle, which is in touch with the extrapallial fluid, of the giant clam Tridacna squamosa. The deduced PMCA sequence consisted of an apical targeting element. Immunofluorescence microscopy confirmed that PMCA had an apical localization in the shell-facing epithelium of the inner mantle, whereby it can actively secrete Ca2+ in exchange for H+. More importantly, the apical PMCA-immunofluorescence of the shell-facing epithelium of the inner mantle increased significantly after 12 h of exposure to light. The transcript and protein levels of PMCA/PMCA also increased significantly in the inner mantle after 6 or 12 h of light exposure. These results offer insights into a light-dependable mechanism of shell formation in T. squamosa and a novel explanation of light-enhanced calcification in general. As the inner mantle normally lacks light sensitive pigments, our results support a previous proposition that symbiotic zooxanthellae, particularly those in the colorful and extensible outer mantle, may act as light-sensing elements for the host clam.

  20. TRPP2 and TRPV4 form an EGF-activated calcium permeable channel at the apical membrane of renal collecting duct cells.

    Directory of Open Access Journals (Sweden)

    Zhi-Ren Zhang

    Full Text Available Regulation of apical calcium entry is important for the function of principal cells of the collecting duct. However, the molecular identity and the regulators of the transporter/channel, which is responsible for apical calcium entry and what factors regulate the calcium conduction remain unclear.We report that endogenous TRPP2 and TRPV4 assemble to form a 23-pS divalent cation-permeable non-selective ion channel at the apical membrane of renal principal cells of the collecting duct. TRPP2\\TRPV4 channel complex was identified by patch-clamp, immunofluorescence and co-immunprecipitation studies in both principal cells that either possess normal cilia (cilia (+ or in which cilia are absent (cilia (-. This channel has distinct biophysical and pharmacological and regulatory profiles compared to either TRPP2 or TRPV4 channels. The rate of occurrence detected by patch clamp was higher in cilia (- compared to cilia (+ cells. In addition, shRNA knockdown of TRPP2 increased the prevalence of TRPV4 channel activity while knockdown of TRPV4 resulted in TRPP2 activity and knockdown of both proteins vastly decreased the 23-pS channel activity. Epidermal growth factor (EGF stimulated TRPP2\\TRPV4 channel through the EGF receptor (EGFR tyrosine kinase-dependent signaling. With loss of cilia, apical EGF treatment resulted in 64-fold increase in channel activity in cilia (- but not cilia (+ cells. In addition EGF increased cell proliferation in cilia (- cell that was dependent upon TRPP2\\TRPV4 channel mediated increase in intracellular calcium.We conclude that in the absence of cilia, an EGF activated TRPP2\\TRPV4 channel may play an important role in increased cell proliferation and cystogenesis.

  1. Subdominant outer membrane antigens in anaplasma marginale: conservation, antigenicity, and protective capacity using recombinant protein

    Science.gov (United States)

    Anaplasma marginale is a tick-borne rickettsial pathogen of cattle with a worldwide distribution. Currently a safe and efficacious vaccine is unavailable. Outer membrane protein (OMP) extracts or a well- defined surface protein complex reproducibly induce protective immunity. However, there are seve...

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

    International Nuclear Information System (INIS)

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

    1986-01-01

    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 1 , C 7 , D 3 , D 7 and H 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 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 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

  3. EBIO, an agent causing maintained epithelial chloride secretion by co-ordinate actions at both apical and basolateral membranes.

    Science.gov (United States)

    MacVinish, L J; Keogh, J; Cuthbert, A W

    2001-01-01

    The effect of 1-ethyl-2-benzimidazolone (EBIO) on electrogenic chloride secretion in murine colonic and nasal epithelium was investigated by the short-circuit technique. In the colon, EBIO produces a sustained current increase in the presence of amiloride, which is sensitive to furosemide. In nasal epithelium EBIO causes only a small, transient current increase. Sustained increases in current were obtained in response to forskolin in both epithelia. To examine the mechanisms by which EBIO increases chloride secretion, the effects on intracellular mediators were measured in colonic crypts. There was no effect on [Ca(2+)]i but cAMP content was increased, more so in the presence of IBMX, indicating a direct effect on adenylate cyclase. In colonic epithelia in which the apical surface was permeabilized by nystatin, and the tissue subjected to an apical to basolateral K(+) gradient, EBIO caused a current increase that was entirely sensitive to charybdotoxin (ChTX). In similarly permeabilized colons Br-cAMP caused a current increase that was entirely sensitive to 293B. Thus EBIO increases chloride secretion in the colon by coordinated actions at both the apical and basolateral faces of the cells. These include direct and indirect actions on Ca(2+)-sensitive and cAMP-sensitive K(+) channels respectively, and indirect actions on the basolateral cotransporter and apical CFTR chloride channels via cAMP. In CF colonic epithelia EBIO did not evoke chloride secretion. It is not clear why the nasal epithelium responds poorly to EBIO whereas it gives a sustained response to the related compound chlorzoxazone.

  4. The outer membrane, not a coat of host proteins, limits antigenicity of virulent Treponema pallidum.

    Science.gov (United States)

    Cox, D L; Chang, P; McDowall, A W; Radolf, J D

    1992-01-01

    Virulent Treponema pallidum reacts poorly with the specific antibodies present in human and rabbit syphilitic sera, a phenomenon often attributed to an outer coat of host serum proteins. Here we present additional evidence that the limited antigenicity of virulent organisms actually is due to a paucity of proteins in the outer membrane. Initially, we used electron microscopy to demonstrate that the outer membrane is highly susceptible to damage from physical manipulation (i.e., centrifugation and resuspension) and nonionic detergents. Organisms with disrupted outer membranes were markedly more antigenic than intact treponemes as determined by immunoelectron microscopy (IEM) with rabbit syphilitic and antiendoflagellar antisera. Data obtained with a new radioimmunoassay, designated the T. pallidum surface-specific radioimmunoassay, corroborated these IEM findings by demonstrating that the major T. pallidum immunogens are not surface exposed; the assay also was unable to detect serum proteins, including fibronectin, on the surfaces of intact organisms. Furthermore, IEM of T. pallidum on ultrathin cryosections with monospecific anti-47-kDa-immunogen antiserum confirmed the intracellular location of the 47-kDa immunogen. On the basis of these and previous findings, we proposed a new model for T. pallidum ultrastructure in which the outer membrane contains a small number of transmembrane proteins and the major membrane immunogens are anchored by lipids to the periplasmic leaflet of the cytoplasmic membrane. This unique ultrastructure explains the remarkable ability of virulent organisms to evade the humoral immune response of the T. pallidum-infected host. Images PMID:1541522

  5. Cancer Patient T Cells Genetically Targeted to Prostate-Specific Membrane Antigen Specifically Lyse Prostate Cancer Cells and Release Cytokines in Response to Prostate-Specific Membrane Antigen

    Directory of Open Access Journals (Sweden)

    Michael C. Gong

    1999-06-01

    Full Text Available The expression of immunoglobulin-based artificial receptors in normal T lymphocytes provides a means to target lymphocytes to cell surface antigens independently of major histocompatibility complex restriction. Such artificial receptors have been previously shown to confer antigen-specific tumoricidal properties in murine T cells. We constructed a novel ζ chain fusion receptor specific for prostate-specific membrane antigen (PSMA termed Pz-1. PSMA is a cell-surface glycoprotein expressed on prostate cancer cells and the neovascular endothelium of multiple carcinomas. We show that primary T cells harvested from five of five patients with different stages of prostate cancer and transduced with the Pz-1 receptor readily lyse prostate cancer cells. Having established a culture system using fibroblasts that express PSMA, we next show that T cells expressing the Pz-1 receptor release cytokines in response to cell-bound PSMA. Furthermore, we show that the cytokine release is greatly augmented by B7.1-mediated costimulation. Thus, our findings support the feasibility of adoptive cell therapy by using genetically engineered T cells in prostate cancer patients and suggest that both CD4+ and CD8+ T lymphocyte functions can be synergistically targeted against tumor cells.

  6. (Na+ + K+)-ATPase and plasma membrane polarity of intestinal epithelial cells: Presence of a brush border antigen in the distal large intestine that is immunologically related to beta subunit

    Energy Technology Data Exchange (ETDEWEB)

    Marxer, A.; Stieger, B.; Quaroni, A.; Kashgarian, M.; Hauri, H.P. (Univ. of Basel (Switzerland))

    1989-09-01

    The previously produced monoclonal antibody IEC 1/48 against cultured rat intestinal crypt cells was extensively characterized and found to be directed against the beta subunit of (Na+ + K+)-ATPase as assessed by immunological and enzymatic criteria. Under nondenaturing conditions the antibody precipitated the alpha-beta enzyme complex (98,000 and 48,000 Mr). This probe, together with the monoclonal antibody C 62.4 against the alpha subunit was used to localize (Na+ + K+)-ATPase in epithelial cells along the rat intestinal tract by immunofluorescence and immunoelectron microscopy. Both antibodies exclusively labeled the basolateral membrane of small intestine and proximal colon epithelial cells. However, in the distal colon, IEC 1/48, but not C 62.4, also labeled the brush border membrane. The cross-reacting beta-subunit-like antigen on the apical cell pole was tightly associated with isolated brush borders but was apparently devoid of (Na+ + K+)-ATPase activity. Subcellular fractionation of colonocytes in conjunction with limited proteolysis and surface radioiodination of intestinal segments suggested that the cross-reacting antigen in the brush border may be very similar to the beta subunit. The results support the notion that in the small intestine and proximal colon the enzyme subunits are exclusively targeted to the basolateral membrane while in the distal colon nonassembled beta subunit or a beta-subunit-like protein is also transported to the apical cell pole.

  7. MHC I presentation of Toxoplasma gondii immunodominant antigen does not require Sec22b and is regulated by antigen orientation at the vacuole membrane.

    Science.gov (United States)

    Buaillon, Célia; Guerrero, Nestor A; Cebrian, Ignacio; Blanié, Sophie; Lopez, Jodie; Bassot, Emilie; Vasseur, Virginie; Santi-Rocca, Julien; Blanchard, Nicolas

    2017-07-01

    The intracellular Toxoplasma gondii parasite replicates within a parasitophorous vacuole (PV). T. gondii secretes proteins that remain soluble in the PV space, are inserted into PV membranes or are exported beyond the PV boundary. In addition to supporting T. gondii growth, these proteins can be processed and presented by MHC I for CD8 + T-cell recognition. Yet it is unclear whether membrane binding influences the processing pathways employed and if topology of membrane antigens impacts their MHC I presentation. Here we report that the MHC I pathways of soluble and membrane-bound antigens differ in their requirement for host ER recruitment. In contrast to the soluble SAG1-OVA model antigen, we find that presentation of the membrane-bound GRA6 is independent from the SNARE Sec22b, a key molecule for transfer of host endoplasmic reticulum components onto the PV. Using parasites modified to secrete a transmembrane antigen with opposite orientations, we further show that MHC I presentation is highly favored when the C-terminal epitope is exposed to the host cell cytosol, which corresponds to GRA6 natural orientation. Our data suggest that the biochemical properties of antigens released by intracellular pathogens critically guide their processing pathway and are valuable parameters to consider for vaccination strategies. © 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  8. Outer membrane proteins analysis of Shigella sonnei and evaluation of their antigenicity in Shigella infected individuals.

    Directory of Open Access Journals (Sweden)

    Hemavathy Harikrishnan

    Full Text Available Bacillary dysentery caused by infection with Shigella spp. remains as serious and common health problem throughout the world. It is a highly multi drug resistant organism and rarely identified from the patient at the early stage of infection. S. sonnei is the most frequently isolated species causing shigellosis in industrialized countries. The antigenicity of outer membrane protein of this pathogen expressed during human infection has not been identified to date. We have studied the antigenic outer membrane proteins expressed by S. sonnei, with the aim of identifying presence of specific IgA and IgG in human serum against the candidate protein biomarkers. Three antigenic OMPs sized 33.3, 43.8 and 100.3 kDa were uniquely recognized by IgA and IgG from patients with S. sonnei infection, and did not cross-react with sera from patients with other types of infection. The antigenic proteome data generated in this study are a first for OMPs of S. sonnei, and they provide important insights of human immune responses. Furthermore, numerous prime candidate proteins were identified which will aid the development of new diagnostic tools for the detection of S. sonnei.

  9. Perbedaan Metode ELISA Sandwich A dan B dalam Deteksi Antigen Membran Toxoplasma gondii

    Directory of Open Access Journals (Sweden)

    ADI PRAYITNO

    2004-11-01

    Full Text Available Spreading of toxoplasmosis to fetus can by placenta, so it caused theabortion, born dead or congenital defect. To diagnosis this disease for fixed the acute infection must get the significant increasing of IgG by the soft fee. The objections of this study are to know the difference between ELISA Sandwich A and B in detecting of membrane antigen of Toxoplasma gondii (T. gondii in placenta tissue of pregnant women three-semester I and II with spontaneous abortion in Surakarta. One hundred serum and placenta tissue samples of pregnant women three-semester I and II with spontaneous abortion are got from dr. Muwardi Hospital. IgM anti Toxo from serum was examined by Toxo ISAGA Kit and IgG anti Toxo by Toxo Screen DA Kit. Detecting of membrane antigen of T. goodie from placenta tissue were done by ELISA Sandwich A and B. The result of this experiment showed that 33% were positive IBM and or Gig anti Toxo. Detection of membrane antigen toward 33 samples with positive Toxo (IgG positive was highly significant different between ELISA Sandwich A (3% positive toward ELISA Sandwich B (72.7% positive.

  10. Calmodulin and CaMKII modulate ENaC activity by regulating the association of MARCKS and the cytoskeleton with the apical membrane.

    Science.gov (United States)

    Alli, Abdel A; Bao, Hui-Fang; Liu, Bing-Chen; Yu, Ling; Aldrugh, Summer; Montgomery, Darrice S; Ma, He-Ping; Eaton, Douglas C

    2015-09-01

    Phosphatidylinositol bisphosphate (PIP2) regulates epithelial sodium channel (ENaC) open probability. In turn, myristoylated alanine-rich C kinase substrate (MARCKS) protein or MARCKS-like protein 1 (MLP-1) at the plasma membrane regulates the delivery of PIP2 to ENaC. MARCKS and MLP-1 are regulated by changes in cytosolic calcium; increasing calcium promotes dissociation of MARCKS from the membrane, but the calcium-regulatory mechanisms are unclear. However, it is known that increased intracellular calcium can activate calmodulin and we show that inhibition of calmodulin with calmidazolium increases ENaC activity presumably by regulating MARCKS and MLP-1. Activated calmodulin can regulate MARCKS and MLP-1 in two ways. Calmodulin can bind to the effector domain of MARCKS or MLP-1, inactivating both proteins by causing their dissociation from the membrane. Mutations in MARCKS that prevent calmodulin association prevent dissociation of MARCKS from the membrane. Calmodulin also activates CaM kinase II (CaMKII). An inhibitor of CaMKII (KN93) increases ENaC activity, MARCKS association with ENaC, and promotes MARCKS movement to a membrane fraction. CaMKII phosphorylates filamin. Filamin is an essential component of the cytoskeleton and promotes association of ENaC, MARCKS, and MLP-1. Disruption of the cytoskeleton with cytochalasin E reduces ENaC activity. CaMKII phosphorylation of filamin disrupts the cytoskeleton and the association of MARCKS, MLP-1, and ENaC, thereby reducing ENaC open probability. Taken together, these findings suggest calmodulin and CaMKII modulate ENaC activity by destabilizing the association between the actin cytoskeleton, ENaC, and MARCKS, or MLP-1 at the apical membrane. Copyright © 2015 the American Physiological Society.

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

    Science.gov (United States)

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

    1986-10-01

    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 globular domain (NC1) of type IV collagen from normal human GBM, while FNS identified only the 26,000-mol wt monomer. FNS reacted with EBM of 12 controls and nine unaffected male kindred members but not EBM of eight affected males. Five affected females exhibited interrupted reactivity of FNS with EBM. GPS showed variable reactivity with EBM and was not discriminating with respect to Alport-type FN. FNS did not stain renal basement members of five affected males. However, the EBM, tubular basement membrane, and Bowman's capsules of affected males contained antigens reactive with GPS. These immunochemical studies suggest that the FNS antigen is distinct from Goodpasture antigen(s). The expression of FNS antigen located on the NC1 domain of type IV collagen is altered in basement membranes of patients with Alport-type FN, and the distribution of this antigenic anomaly within kindreds suggests X-linked dominant transmission of a defective gene.

  12. Comparative analysis of monoclonal antibodies against prostate-specific membrane antigen (PSMA)

    Czech Academy of Sciences Publication Activity Database

    Tykvart, Jan; Navrátil, Václav; Sedlák, František; Corey, E.; Colombatti, M.; Fracasso, G.; Koukolík, F.; Bařinka, Cyril; Šácha, Pavel; Konvalinka, Jan

    2014-01-01

    Roč. 74, č. 16 (2014), s. 1674-1690 ISSN 0270-4137 R&D Projects: GA ČR GAP304/12/0847; GA MŠk LO1302; GA ČR GAP301/12/1513; GA MŠk(CZ) ED1.1.00/02.0109 Institutional support: RVO:61388963 ; RVO:86652036 Keywords : glutamate carboxypeptidase II * prostate-specific membrane antigen * folate hydrolase * NAALADase * Western blot * immunohistochemistry * ELISA * flow cytometry * surface plasmon resonance Subject RIV: CE - Biochemistry Impact factor: 3.565, year: 2014

  13. Calcium Binding by Ro 60 Multiple Antigenic Peptides on PVDF Membrane.

    Science.gov (United States)

    Kurien, Biji T; Bachmann, Michael P

    2015-01-01

    Antibodies directed against ribonucleoprotein (RNP) particles are observed in systemic lupus erythematosus. Ro RNP particle is one such target. It is composed of a 60 kDa protein (Ro 60 or SS-A) that is non-covalently associated with at least one of the four short uridine-rich RNAs (the hY RNAs). Previously, we showed that multiple antigenic peptides (MAPs) made from the sequence of the Ro 60 autoantigen could be used, using double-immunodiffusion studies, enzyme-linked immunosorbant assay, affinity chromatography, and surface plasmon resonance, to show intramolecular and intermolecular protein-protein interaction within the Ro 60 RNP particle. We also observed that calcium is important in mediating this interaction. We hypothesized, therefore, that 60 kDa Ro is a calcium-binding protein. To investigate this, we electrophoresed 60 kDa Ro MAPs, transferred them to PVDF membrane, and assayed calcium binding using the Quin-2 system. Several Ro 60 MAPs were found to bind calcium using this assay, as well as bovine serum albumin, another calcium-binding protein. However, a MAP constructed from the Sm autoantigen did not bind to calcium. These data, along with our observation regarding the involvement of calcium in protein-protein interaction occurring between Ro 60 antigen and Ro 60 MAPs, makes us propose that Ro 60 antigen is a calcium-binding protein.

  14. Comparison of five assays for antibody to varicella-zoster virus and the fluorescent-antibody-to-membrane-antigen test.

    OpenAIRE

    Larussa, P; Steinberg, S; Waithe, E; Hanna, B; Holzman, R

    1987-01-01

    Three commercially available assays (the Varicelisa Test Kit [Whittaker M.A. Bioproducts, Walkersville, Md.], the VZV Indirect Fluorescent-Antibody Test [Electro-Nucleonics, Inc., Columbia, Md.], and the Litton VZV Bio-EnzaBead Screen Kit [Litton Bionetics, Inc., Charleston, S.C.]) and two enzyme-linked immunosorbent assays used in our laboratory, one using a membrane-associated antigen and the other using a soluble antigen dotted on nitrocellulose paper, were compared with a varicella-zoster...

  15. Organic Cation Transporter 1 (OCT1/mOct1) Is Localized in the Apical Membrane of Caco-2 Cell Monolayers and Enterocytes

    Science.gov (United States)

    Han, Tianxiang (Kevin); Everett, Ruth S.; Proctor, William R.; Ng, Chee M.; Costales, Chester L.; Brouwer, Kim L. R.

    2013-01-01

    Organic cation transporters (OCTs) are members of the solute carrier 22 family of transporter proteins that are involved in absorption, distribution, and excretion of organic cations. OCT3 is localized in the apical (AP) membrane of enterocytes, but the literature is ambiguous about OCT1 (mOct1) localization, with some evidence suggesting a basolateral (BL) localization in human and mouse enterocytes. This is contrary to our preliminary findings showing AP localization of OCT1 in Caco-2 cell monolayers, an established model of human intestinal epithelium. Therefore, this study aims at determining the localization of OCT1 (mOct1) in Caco-2 cells, and human and mouse enterocytes. Functional studies using OCT1-specific substrate pentamidine showed transporter-mediated AP but not BL uptake in Caco-2 cells and human and mouse intestinal tissues. OCT1 inhibition decreased AP uptake of pentamidine by ∼50% in all three systems with no effect on BL uptake. A short hairpin RNA-mediated OCT1 knockdown in Caco-2 cells decreased AP uptake of pentamidine by ∼50% but did not alter BL uptake. Immunostaining and confocal microscopy in all three systems confirmed AP localization of OCT1 (mOct1). Our studies unequivocally show AP membrane localization of OCT1 (mOct1) in Caco-2 cells and human and mouse intestine. These results are highly significant as they will require reinterpretation of previous drug disposition and drug-drug interaction studies where conclusions were drawn assuming BL localization of OCT1 in enterocytes. Most importantly, these results will require revision of the regulatory guidance for industry in the United States and elsewhere because it has stated that OCT1 is basolaterally localized in enterocytes. PMID:23680637

  16. Caprine nictitating membrane cell culture for production of small ruminants lentivirus antigen

    Directory of Open Access Journals (Sweden)

    Dalva Alana Aragão de Azevedo

    2015-12-01

    Full Text Available ABSTRACT. Azevedo D.A.A., Araújo J.F., Sousa A.L.M. de, Dias R.P., Souza T.S. de, Andrioli A. & Pinheiro R.R. [Caprine nictitating membrane cell culture for production of small ruminants lentivirus antigen.] Produção de antí- geno de lentivírus de pequenos ruminantes através da cultura celular de membrana nictitante caprina. Revista Brasileira de Medicina Veterinária, 37(4:316-320, 2015. Empresa Brasileira de Pesquisa Agropecuária (Embrapa Caprinos e Ovinos, Fazenda Três Lagoas, Estrada Sobral/Jordão Km 4, Sobral, CE 62010-970, Brasil. E-mail: dalvaazevedo@outlook.com Small Ruminants lentivirus (SRLV belong to the Retroviridae family and is the etiologic agent of caprine arthritis encephalitis in goats and maedi-visna in sheep. Cells of the monocytic-phagocytic lineage are the main viral targets and infected hosts cannot develop a curative immune response. We evaluated whether nictitating membrane (NM cells can be cultivated in vitro and also whether it is a feasible system for SRLV antigen production for immunodiagnosis purposes. MN cells were collected from a SRLV negative animal and cultivated in minimal essential media supplemented with bovine fetal serum. Subcultures were inoculated with caprine arthritis encephalitis virus (CAEV Cork strain, the supernatant was collected, clarified by centrifugation (3.400 g for 20 minutes, concentrated using the AMICON® system. Samples were subjected to agar gel immunodiffusion (AGID tests. MN cells presented satisfactory performance, were amenable to several subcultivation.

  17. Advances in prostate-specific membrane antigen PET of prostate cancer.

    Science.gov (United States)

    Bouchelouche, Kirsten; Choyke, Peter L

    2018-05-01

    In recent years, a large number of reports have been published on prostate-specific membrane antigen (PSMA)/PET in prostate cancer (PCa). This review highlights advances in PSMA PET in PCa during the past year. PSMA PET/computed tomography (CT) is useful in detection of biochemical recurrence, especially at low prostate-specific antigen (PSA) values. The detection rate of PSMA PET is influenced by PSA level. For primary PCa, PSMA PET/CT shows promise for tumour localization in the prostate, especially in combination with multiparametric MRI (mpMRI). For primary staging, PSMA PET/CT can be used in intermediate and high-risk PCa. Intraoperative PSMA radioligand guidance seems promising for detection of malignant lymph nodes. While the use of PSMA PET/MRI in primary localized disease is limited to high and intermediate-risk patients and localized staging, in the recurrence setting, PET/MRI can be particularly helpful when the lesions are subtle. PSMA PET/CT is superior to choline PET/CT and other conventional imaging modalities. Molecular imaging with PSMA PET continues to pave the way for personalized medicine in PCa.However, large prospective clinical studies are still needed to fully evaluate the role of PSMA PET/CT and PET/MRI in the clinical workflow of PCa.

  18. A remote arene-binding site on prostate specific membrane antigen revealed by antibody-recruiting small molecules

    Czech Academy of Sciences Publication Activity Database

    Zhang, A.X.; Murelli, R.P.; Bařinka, Cyril; Michel, J.; Cocleaza, A.; Jorgensen, W.L.; Lubkowski, J.; Spiegel, D.A.

    2010-01-01

    Roč. 132, č. 36 (2010), s. 12711-12716 ISSN 0002-7863 Institutional research plan: CEZ:AV0Z50520701 Keywords : Prostate -specific membrane antigen * antibody recruiting molecules * Structure-activity relationship Subject RIV: CE - Biochemistry Impact factor: 9.019, year: 2010

  19. Discriminatory Role of Detergent-Resistant Membranes in the Dimerization and Endocytosis of Prostate-Specific Membrane Antigen.

    Directory of Open Access Journals (Sweden)

    Sonja Schmidt

    Full Text Available Prostate-specific membrane antigen (PSMA is a type-II membrane glycoprotein that was initially identified in LNCaP cells. It is expressed at elevated levels in prostate cancer. In view of the correlation between the expression levels of PSMA and disease grade and stage, PSMA is considered to be one of the most promising biomarkers in the diagnosis and treatment of prostate cancer. In LNCaP cells PSMA undergoes internalization via clathrin-coated pits followed by accumulation in the endosomes. PSMA associates with different types of detergent-resistant membranes (DRMs along the secretory pathway. Its mature form is mainly insoluble in Lubrol WX, but does not associate with Triton X-100-DRMs. To understand the mechanism of PSMA internalization we investigated its association during internalization with DRMs. For this purpose, internalization was induced by antibody cross-linking. We demonstrate at the biochemical and cell biological levels that: [i] exclusively homodimers of PSMA are associated with Lubrol WX-DRMs, [ii] antibody-induced cross-linking of PSMA molecules results in a time-dependent partitioning into another DRMs type, namely Triton X-100-DRMs, and [iii] concomitant with its association with Triton-X-100-DRMs internalization of PSMA occurs along tubulin filaments. In a previous work (Colombatti et al. (2009 PLoS One 4: e4608 we demonstrated that the small GTPases RAS and RAC1 and the MAPKs p38 and ERK1/2 are activated during antibody cross-linking. As downstream effects of this activation we observed a strong induction of NF-kB associated with an increased expression of IL-6 and CCL5 genes and that IL-6 and CCL5 enhanced the proliferative potential of LNCaP cells synergistically. These observations together with findings reported here hypothesize a fundamental role of DRMs during activation of PSMA as platforms for trafficking, endocytosis and signalling. Understanding these mechanisms constitutes an essential prerequisite for

  20. Discriminatory Role of Detergent-Resistant Membranes in the Dimerization and Endocytosis of Prostate-Specific Membrane Antigen

    Science.gov (United States)

    Schmidt, Sonja; Gericke, Birthe; Fracasso, Giulio; Ramarli, Dunia; Colombatti, Marco; Naim, Hassan Y.

    2013-01-01

    Prostate-specific membrane antigen (PSMA) is a type-II membrane glycoprotein that was initially identified in LNCaP cells. It is expressed at elevated levels in prostate cancer. In view of the correlation between the expression levels of PSMA and disease grade and stage, PSMA is considered to be one of the most promising biomarkers in the diagnosis and treatment of prostate cancer. In LNCaP cells PSMA undergoes internalization via clathrin-coated pits followed by accumulation in the endosomes. PSMA associates with different types of detergent-resistant membranes (DRMs) along the secretory pathway. Its mature form is mainly insoluble in Lubrol WX, but does not associate with Triton X-100-DRMs. To understand the mechanism of PSMA internalization we investigated its association during internalization with DRMs. For this purpose, internalization was induced by antibody cross-linking. We demonstrate at the biochemical and cell biological levels that: [i] exclusively homodimers of PSMA are associated with Lubrol WX-DRMs, [ii] antibody-induced cross-linking of PSMA molecules results in a time-dependent partitioning into another DRMs type, namely Triton X-100-DRMs, and [iii] concomitant with its association with Triton-X-100-DRMs internalization of PSMA occurs along tubulin filaments. In a previous work (Colombatti et al. (2009) PLoS One 4: e4608) we demonstrated that the small GTPases RAS and RAC1 and the MAPKs p38 and ERK1/2 are activated during antibody cross-linking. As downstream effects of this activation we observed a strong induction of NF-kB associated with an increased expression of IL-6 and CCL5 genes and that IL-6 and CCL5 enhanced the proliferative potential of LNCaP cells synergistically. These observations together with findings reported here hypothesize a fundamental role of DRMs during activation of PSMA as platforms for trafficking, endocytosis and signalling. Understanding these mechanisms constitutes an essential prerequisite for utilization of PSMA as

  1. FLUORESCENCE OVERLAY ANTIGEN MAPPING OF THE EPIDERMAL BASEMENT-MEMBRANE ZONE .2. COLOR FIDELITY

    NARCIS (Netherlands)

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

    In this second report on the fluorescence overlay antigen mapping (FOAM) technique, we highlight some of the errors that may influence faithful color rendition of slide preparations using triple antigen immunofluorescence staining. Reliable interpretation of multicolor fluorescence images requires

  2. Promotion of arthritis and allergy in mice by aminoglycoglycerophospholipid, a membrane antigen specific to Mycoplasma fermentans.

    Science.gov (United States)

    Sato, Naoki; Oizumi, Takefumi; Kinbara, Masayuki; Sato, Tadasu; Funayama, Hiromi; Sato, Seiji; Matsuda, Kazuhiro; Takada, Haruhiko; Sugawara, Shunji; Endo, Yasuo

    2010-06-01

    Mycoplasmas, which lack a cell wall and are the smallest self-replicating bacteria, have been linked to some chronic diseases, such as AIDS, rheumatoid arthritis (RA), and oncogenic transformation of cells. Their membrane components (lipoproteins and glycolipids) have been identified as possible causative factors in such diseases. Glycoglycerophospholipid (GGPL)-III, a unique phosphocholine-containing aminoglycoglycerophospholipid, is a major specific antigen of Mycoplasma fermentans, and has been detected in 38% of RA patients. Unlike those of lipoproteins, which induce inflammation via Toll-like receptor 2 (TLR2), the pathologic effects of GGPL-III are poorly understood. RA and metal allergies are chronic inflammatory diseases in which autoantigens have been implicated. Here, we examined the effects of chemically synthesized GGPL-III in murine arthritis and allergy models. GGPL-III alone exhibited little inflammatory effect, but promoted both collagen-induced arthritis and nickel (Ni) allergy, although less powerfully than Escherichia coli lipopolysaccharide. The augmenting effect of GGPL-III on Ni allergy was present in mice deficient in either T cells or active TLR4, but it was markedly weaker in mice deficient in macrophages, interleukin-1, or the histamine-forming enzyme histidine decarboxylase than in their control strains. These results suggest that GGPL-III may play roles in some types of chronic diseases via the innate immune system.

  3. Evaluation of vaginal fluid squamous cell carcinoma antigen test in diagnosis of premature rupture of membranes.

    Science.gov (United States)

    Koike, Natsuki; Oi, Hidekazu; Naruse, Katsuhiko; Tsunemi, Taihei; Akasaka, Juria; Sado, Toshiyuki; Kobayashi, Hiroshi

    2017-02-01

    Previous studies have reported that concentrations of squamous cell carcinoma (SCC) antigen in amniotic fluid are extremely higher than that in the maternal serum. The aim of this study was to assess the potential clinical utility of vaginal fluid SCC level as a marker for diagnosing premature rupture of membranes (PROM). A case-control study was performed using patients admitted to Nara Medical University Hospital, delivery ward, from January 2011 to December 2012. The discriminatory potential of SCC assay was determined using 54 PROM and 108 gestational age-matched control vaginal fluid samples, in a 1:2 ratio. Levels of vaginal fluid SCC in patients with PROM and control pregnant women were quantified by an enzyme-linked immunosorbent assay. The statistical results showed no correlation between gestational age and vaginal fluid SCC levels. There was no significant difference in vaginal fluid SCC levels between patients with PROM and those with control pregnant women (16156.5 ± 10495.8 ng/mL versus 15471.9 ± 11362.2 ng/mL, p = 0.467). We conclude that SCC could not be regarded as a potential marker for diagnosis of PROM. SCC may be a physiologic constituent of the vaginal fluid during pregnancy.

  4. Role of the major antigenic membrane protein in phytoplasma transmission by two insect vector species.

    Science.gov (United States)

    Rashidi, Mahnaz; Galetto, Luciana; Bosco, Domenico; Bulgarelli, Andrea; Vallino, Marta; Veratti, Flavio; Marzachì, Cristina

    2015-09-30

    Phytoplasmas are bacterial plant pathogens (class Mollicutes), transmitted by phloem feeding leafhoppers, planthoppers and psyllids in a persistent/propagative manner. Transmission of phytoplasmas is under the control of behavioral, environmental and geographical factors, but molecular interactions between membrane proteins of phytoplasma and vectors may also be involved. The aim of the work was to provide experimental evidence that in vivo interaction between phytoplasma antigenic membrane protein (Amp) and vector proteins has a role in the transmission process. In doing so, we also investigated the topology of the interaction at the gut epithelium and at the salivary glands, the two barriers encountered by the phytoplasma during vector colonization. Experiments were performed on the 'Candidatus Phytoplasma asteris' chrysanthemum yellows strain (CYP), and the two leafhopper vectors Macrosteles quadripunctulatus Kirschbaum and Euscelidius variegatus Kirschbaum. To specifically address the interaction of CYP Amp at the gut epithelium barrier, insects were artificially fed with media containing either the recombinant phytoplasma protein Amp, or the antibody (A416) or both, and transmission, acquisition and inoculation efficiencies were measured. An abdominal microinjection protocol was employed to specifically address the interaction of CYP Amp at the salivary gland barrier. Phytoplasma suspension was added with Amp or A416 or both, injected into healthy E. variegatus adults and then infection and inoculation efficiencies were measured. An internalization assay was developed, consisting of dissected salivary glands from healthy E. variegatus exposed to phytoplasma suspension alone or together with A416 antibody. The organs were then either observed in confocal microscopy or subjected to DNA extraction and phytoplasma quantification by qPCR, to visualize and quantify possible differences among treatments in localization/presence/number of CYP cells. Artificial feeding

  5. 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 ...

  6. Development of a PET Prostate-Specific Membrane Antigen Imaging Agent: Preclinical Translation for Future Clinical Application

    Science.gov (United States)

    2017-10-01

    Award Number: W81XWH-14-1-0603 TITLE: Development of a PET Prostate-Specific Membrane Antigen Imaging Agent: Preclinical Translation for Future...other documentation. REPORT DOCUMENTATION PAGE Form Approved OMB No. 0704-0188 Public reporting burden for this collection of information is estimated...needed, and completing and reviewing this collection of information. Send comments regarding this burden estimate or any other aspect of this

  7. Tissue expression and enzymologic characterization of human prostate specific membrane antigen and its rat and pig orthologs

    Czech Academy of Sciences Publication Activity Database

    Rovenská, Miroslava; Hlouchová, Klára; Šácha, Pavel; Mlčochová, Petra; Horák, Vratislav; Zámečník, J.; Bařinka, C.; Konvalinka, Jan

    2008-01-01

    Roč. 68, č. 2 (2008), s. 171-182 ISSN 0270-4137 R&D Projects: GA MŠk 1M0508; GA ČR GA524/04/0102 Institutional research plan: CEZ:AV0Z40550506; CEZ:AV0Z50450515 Keywords : prostate specific membrane antigen * glutamate carboxypeptidase II * animal orthologs * prostate cancer * animal model Subject RIV: CE - Biochemistry Impact factor: 3.069, year: 2008

  8. Prostate-Specific Membrane Antigen Targeted Gold Nanoparticles for Theranostics of Prostate Cancer.

    Science.gov (United States)

    Mangadlao, Joey Dacula; Wang, Xinning; McCleese, Christopher; Escamilla, Maria; Ramamurthy, Gopalakrishnan; Wang, Ziying; Govande, Mukul; Basilion, James P; Burda, Clemens

    2018-04-24

    Prostate cancer is one of the most common cancers and among the leading causes of cancer deaths in the United States. Men diagnosed with the disease typically undergo radical prostatectomy, which often results in incontinence and impotence. Recurrence of the disease is often experienced by most patients with incomplete prostatectomy during surgery. Hence, the development of a technique that will enable surgeons to achieve a more precise prostatectomy remains an open challenge. In this contribution, we report a theranostic agent (AuNP-5kPEG-PSMA-1-Pc4) based on prostate-specific membrane antigen (PSMA-1)-targeted gold nanoparticles (AuNPs) loaded with a fluorescent photodynamic therapy (PDT) drug, Pc4. The fabricated nanoparticles are well-characterized by spectroscopic and imaging techniques and are found to be stable over a wide range of solvents, buffers, and media. In vitro cellular uptake experiments demonstrated significantly higher nanoparticle uptake in PSMA-positive PC3pip cells than in PSMA-negative PC3flu cells. Further, more complete cell killing was observed in Pc3pip than in PC3flu cells upon exposure to light at different doses, demonstrating active targeting followed by Pc4 delivery. Likewise, in vivo studies showed remission on PSMA-expressing tumors 14 days post-PDT. Atomic absorption spectroscopy revealed that targeted AuNPs accumulate 4-fold higher in PC3pip than in PC3flu tumors. The nanoparticle system described herein is envisioned to provide surgical guidance for prostate tumor resection and therapeutic intervention when surgery is insufficient.

  9. Antigen sequence typing of outer membrane protein (fetA gene of Neisseria meningitidis serogroup A from Delhi & adjoining areas

    Directory of Open Access Journals (Sweden)

    S Dwivedi

    2014-01-01

    Full Text Available Background & objectives: Meningitis caused by Neisseria meningitidis is a fatal disease. Meningococcal meningitis is an endemic disease in Delhi and irregular pattern of outbreaks has been reported in India. All these outbreaks were associated with serogroup A. Detailed molecular characterization of N. meningitidis is required for the management of this fatal disease. In this study, we characterized antigenic diversity of surface exposed outer membrane protein (OMP FetA antigen of N. meningitidis serogroup A isolates obtained from cases of invasive meningococcal meningitis in Delhi, India. Methods: Eight isolates of N. meningitidis were collected from cerebrospinal fluid during October 2008 to May 2011 from occasional cases of meningococcal meningitis. Seven isolates were from outbreaks of meningococcal meningitis in 2005-2006 in Delhi and its adjoining areas. These were subjected to molecular typing of fetA gene, an outer membrane protein gene. Results: All 15 N. meningitides isolates studied were serogroup A. This surface exposed porin is putatively under immune pressure. Hence as a part of molecular characterization, genotyping was carried out to find out the diversity in outer membrane protein (FetA gene among the circulating isolates of N. meningitidis. All 15 isolates proved to be of the same existing allele type of FetA variable region (VR when matched with global database. The allele found was F3-1 for all the isolates. Interpretation & conclusions: There was no diversity reported in the outer membrane protein FetA in the present study and hence this protein appeared to be a stable molecule. More studies on molecular characterization of FetA antigen are required from different serogroups circulating in different parts of the world.

  10. 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

    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

  11. Label-free quantitative mass spectrometry for analysis of protein antigens in a meningococcal group B outer membrane vesicle vaccine.

    Science.gov (United States)

    Dick, Lawrence W; Mehl, John T; Loughney, John W; Mach, Anna; Rustandi, Richard R; Ha, Sha; Zhang, Lan; Przysiecki, Craig T; Dieter, Lance; Hoang, Van M

    2015-01-01

    The development of a multivalent outer membrane vesicle (OMV) vaccine where each strain contributes multiple key protein antigens presents numerous analytical challenges. One major difficulty is the ability to accurately and specifically quantitate each antigen, especially during early development and process optimization when immunoreagents are limited or unavailable. To overcome this problem, quantitative mass spectrometry methods can be used. In place of traditional mass assays such as enzyme-linked immunosorbent assays (ELISAs), quantitative LC-MS/MS using multiple reaction monitoring (MRM) can be used during early-phase process development to measure key protein components in complex vaccines in the absence of specific immunoreagents. Multiplexed, label-free quantitative mass spectrometry methods using protein extraction by either detergent or 2-phase solvent were developed to quantitate levels of several meningococcal serogroup B protein antigens in an OMV vaccine candidate. Precision was demonstrated to be less than 15% RSD for the 2-phase extraction and less than 10% RSD for the detergent extraction method. Accuracy was 70 to 130% for the method using a 2-phase extraction and 90-110% for detergent extraction. The viability of MS-based protein quantification as a vaccine characterization method was demonstrated and advantages over traditional quantitative methods were evaluated. Implementation of these MS-based quantification methods can help to decrease the development time for complex vaccines and can provide orthogonal confirmation of results from existing antigen quantification techniques.

  12. Erythrocyte Membrane Antigen Frequencies in Patients with Type II Congenital Smell Loss

    Science.gov (United States)

    Stateman, William A.; Henkin, Robert I.; Knöppel, Alexandra; Flegel, Willy A.

    2014-01-01

    OBJECTIVE The objective of this study was to determine whether there are genetic factors associated with Type II congenital smell loss. STUDY DESIGN The expression frequencies of 16 erythrocyte antigens among patients with Type II congenital smell loss were determined and compared to those of a large control group. METHODS Blood samples were obtained from 99 patients with Type II congenital smell loss. Presence of the erythrocyte surface antigens A, B, M, N, S, s, Fya, Fyb, D, C, c, E, e, K, Jka, and Jkb was analyzed by blood group serology. Comparisons of expression frequencies of these antigens were made between the patients and a large control group. RESULTS Patients tested for the Duffy b antigen (Fyb haplotype) exhibited a statistically significant 11% decrease in expression frequency compared to the controls. There were no significant differences between patients and controls in the expression frequencies for all other erythrocyte antigens (A, B, M, N, S, s, Fya, D, C, c, E, e, K, Jka, or Jkb). CONCLUSIONS These findings describe the presence of a previously unrevealed genetic tendency among patients with Type II congenital smell loss related to erythrocyte surface antigen expression. The deviation in expression rate of Duffy b suggests a target gene and chromosome region in which future research into this form of congenital smell loss may reveal a more specific genetic basis for Type II congenital smell loss. PMID:25456515

  13. 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

  14. Exposure to the Epstein–Barr Viral Antigen Latent Membrane Protein 1 Induces Myelin-Reactive Antibodies In Vivo

    Directory of Open Access Journals (Sweden)

    Yakov Lomakin

    2017-07-01

    Full Text Available Multiple sclerosis (MS is an autoimmune chronic inflammatory disease of the central nervous system (CNS. Cross-reactivity of neuronal proteins with exogenous antigens is considered one of the possible mechanisms of MS triggering. Previously, we showed that monoclonal myelin basic protein (MBP-specific antibodies from MS patients cross-react with Epstein–Barr virus (EBV latent membrane protein 1 (LMP1. In this study, we report that exposure of mice to LMP1 results in induction of myelin-reactive autoantibodies in vivo. We posit that chronic exposure or multiple acute exposures to viral antigen may redirect B cells from production of antiviral antibodies to antibodies, specific to myelin antigen. However, even in inbred animals, which are almost identical in terms of their genomes, such an effect is only observed in 20–50% of animals, indicating that this change occurs by chance, rather than systematically. Cross-immunoprecipitation analysis showed that only part of anti-MBP antibodies from LMP1-immunized mice might simultaneously bind LMP1. In contrast, the majority of anti-LMP1 antibodies from MBP-immunized mice bind MBP. De novo sequencing of anti-LMP1 and anti-MBP antibodies by mass spectrometry demonstrated enhanced clonal diversity in LMP1-immunized mice in comparison with MBP-immunized mice. We suggest that induction of MBP-reactive antibodies in LMP1-immunized mice may be caused by either Follicular dendritic cells (FDCs or by T cells that are primed by myelin antigens directly in CNS. Our findings help to elucidate the still enigmatic link between EBV infection and MS development, suggesting that myelin-reactive antibodies raised as a response toward EBV protein LMP1 are not truly cross-reactive but are primarily caused by epitope spreading.

  15. Exposure to the Epstein–Barr Viral Antigen Latent Membrane Protein 1 Induces Myelin-Reactive Antibodies In Vivo

    Science.gov (United States)

    Lomakin, Yakov; Arapidi, Georgii Pavlovich; Chernov, Alexander; Ziganshin, Rustam; Tcyganov, Evgenii; Lyadova, Irina; Butenko, Ivan Olegovich; Osetrova, Maria; Ponomarenko, Natalia; Telegin, Georgy; Govorun, Vadim Markovich; Gabibov, Alexander; Belogurov, Alexey

    2017-01-01

    Multiple sclerosis (MS) is an autoimmune chronic inflammatory disease of the central nervous system (CNS). Cross-reactivity of neuronal proteins with exogenous antigens is considered one of the possible mechanisms of MS triggering. Previously, we showed that monoclonal myelin basic protein (MBP)-specific antibodies from MS patients cross-react with Epstein–Barr virus (EBV) latent membrane protein 1 (LMP1). In this study, we report that exposure of mice to LMP1 results in induction of myelin-reactive autoantibodies in vivo. We posit that chronic exposure or multiple acute exposures to viral antigen may redirect B cells from production of antiviral antibodies to antibodies, specific to myelin antigen. However, even in inbred animals, which are almost identical in terms of their genomes, such an effect is only observed in 20–50% of animals, indicating that this change occurs by chance, rather than systematically. Cross-immunoprecipitation analysis showed that only part of anti-MBP antibodies from LMP1-immunized mice might simultaneously bind LMP1. In contrast, the majority of anti-LMP1 antibodies from MBP-immunized mice bind MBP. De novo sequencing of anti-LMP1 and anti-MBP antibodies by mass spectrometry demonstrated enhanced clonal diversity in LMP1-immunized mice in comparison with MBP-immunized mice. We suggest that induction of MBP-reactive antibodies in LMP1-immunized mice may be caused by either Follicular dendritic cells (FDCs) or by T cells that are primed by myelin antigens directly in CNS. Our findings help to elucidate the still enigmatic link between EBV infection and MS development, suggesting that myelin-reactive antibodies raised as a response toward EBV protein LMP1 are not truly cross-reactive but are primarily caused by epitope spreading. PMID:28729867

  16. Exposure to the Epstein-Barr Viral Antigen Latent Membrane Protein 1 Induces Myelin-Reactive Antibodies In Vivo.

    Science.gov (United States)

    Lomakin, Yakov; Arapidi, Georgii Pavlovich; Chernov, Alexander; Ziganshin, Rustam; Tcyganov, Evgenii; Lyadova, Irina; Butenko, Ivan Olegovich; Osetrova, Maria; Ponomarenko, Natalia; Telegin, Georgy; Govorun, Vadim Markovich; Gabibov, Alexander; Belogurov, Alexey

    2017-01-01

    Multiple sclerosis (MS) is an autoimmune chronic inflammatory disease of the central nervous system (CNS). Cross-reactivity of neuronal proteins with exogenous antigens is considered one of the possible mechanisms of MS triggering. Previously, we showed that monoclonal myelin basic protein (MBP)-specific antibodies from MS patients cross-react with Epstein-Barr virus (EBV) latent membrane protein 1 (LMP1). In this study, we report that exposure of mice to LMP1 results in induction of myelin-reactive autoantibodies in vivo . We posit that chronic exposure or multiple acute exposures to viral antigen may redirect B cells from production of antiviral antibodies to antibodies, specific to myelin antigen. However, even in inbred animals, which are almost identical in terms of their genomes, such an effect is only observed in 20-50% of animals, indicating that this change occurs by chance, rather than systematically. Cross-immunoprecipitation analysis showed that only part of anti-MBP antibodies from LMP1-immunized mice might simultaneously bind LMP1. In contrast, the majority of anti-LMP1 antibodies from MBP-immunized mice bind MBP. De novo sequencing of anti-LMP1 and anti-MBP antibodies by mass spectrometry demonstrated enhanced clonal diversity in LMP1-immunized mice in comparison with MBP-immunized mice. We suggest that induction of MBP-reactive antibodies in LMP1-immunized mice may be caused by either Follicular dendritic cells (FDCs) or by T cells that are primed by myelin antigens directly in CNS. Our findings help to elucidate the still enigmatic link between EBV infection and MS development, suggesting that myelin-reactive antibodies raised as a response toward EBV protein LMP1 are not truly cross-reactive but are primarily caused by epitope spreading.

  17. Quantitative proteomic analysis of Shigella flexneri and Shigella sonnei Generalized Modules for Membrane Antigens (GMMA) reveals highly pure preparations.

    Science.gov (United States)

    Maggiore, Luana; Yu, Lu; Omasits, Ulrich; Rossi, Omar; Dougan, Gordon; Thomson, Nicholas R; Saul, Allan; Choudhary, Jyoti S; Gerke, Christiane

    2016-02-01

    Outer membrane blebs are naturally shed by Gram-negative bacteria and are candidates of interest for vaccines development. Genetic modification of bacteria to induce hyperblebbing greatly increases the yield of blebs, called Generalized Modules for Membrane Antigens (GMMA). The composition of the GMMA from hyperblebbing mutants of Shigella flexneri 2a and Shigella sonnei were quantitatively analyzed using high-sensitivity mass spectrometry with the label-free iBAQ procedure and compared to the composition of the solubilized cells of the GMMA-producing strains. There were 2306 proteins identified, 659 in GMMA and 2239 in bacteria, of which 290 (GMMA) and 1696 (bacteria) were common to both S. flexneri 2a and S. sonnei. Predicted outer membrane and periplasmic proteins constituted 95.7% and 98.7% of the protein mass of S. flexneri 2a and S. sonnei GMMA, respectively. Among the remaining proteins, small quantities of ribosomal proteins collectively accounted for more than half of the predicted cytoplasmic protein impurities in the GMMA. In GMMA, the outer membrane and periplasmic proteins were enriched 13.3-fold (S. flexneri 2a) and 8.3-fold (S. sonnei) compared to their abundance in the parent bacteria. Both periplasmic and outer membrane proteins were enriched similarly, suggesting that GMMA have a similar surface to volume ratio as the surface to periplasmic volume ratio in these mutant bacteria. Results in S. flexneri 2a and S. sonnei showed high reproducibility indicating a robust GMMA-producing process and the low contamination by cytoplasmic proteins support the use of GMMA for vaccines. Data are available via ProteomeXchange with identifier PXD002517. Copyright © 2015 The Authors. Published by Elsevier GmbH.. All rights reserved.

  18. Concentration of membrane antigens by forward transport and trapping in neuronal growth cones.

    Science.gov (United States)

    Sheetz, M P; Baumrind, N L; Wayne, D B; Pearlman, A L

    1990-04-20

    Formation of the nervous system requires that neuronal growth cones follow specific paths and then stop at recognition signals, sensed at the growth cone's leading edge. We used antibody-coated gold particles viewed by video-enhanced differential interference contrast microscopy to observe the distribution and movement of two cell surface molecules, N-CAM and the 2A1 antigen, on growth cones of cultured cortical neurons. Gold particles are occasionally transported forward at 1-2 microns/s to the leading edge where they are trapped but continue to move. Concentration at the edge persists after cytochalasin D treatment or ATP depletion, but active movements to and along edges cease. We also observed a novel outward movement of small cytoplasmic aggregates at 1.8 microns/s in filopodia. We suggest that active forward transport and trapping involve reversible attachment of antigens to and transport along cytoskeletal elements localized to edges of growth cones.

  19. Heterobivalent Imaging Agents for Simultaneous Targeting Prostate-Specific Membrane Antigen (PSMA) and Hepsin

    Science.gov (United States)

    2014-11-01

    rectal examination (DRE) have advanced significantly for the early diagnosis of patients with PCa, the controversy on the efficacy of PSA testing for...Cancer Society (www.cancer.org). Even if current prostate cancer screening methods with the prostate-specific antigen (PSA) blood test and digital ...respectively, indicating that 13 is more hydrophilic than 12. ESI-MS showed [M+H]2+ and [M+H]+ ion peaks of 12 and 13 in the positive modes as shown in

  20. 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......-fluorescence microscopy was used to visualize antibody-mediated adhesion between the surfaces of live infected erythrocytes and atomic force microscopy was used to obtain higher resolution images of the membrane knobs on the infected erythrocyte to estimate knob surface areas and model VAR2CSA packing density on the knob...

  1. 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...

  2. The major antigenic membrane protein of "Candidatus Phytoplasma asteris" selectively interacts with ATP synthase and actin of leafhopper vectors.

    Directory of Open Access Journals (Sweden)

    Luciana Galetto

    Full Text Available Phytoplasmas, uncultivable phloem-limited phytopathogenic wall-less bacteria, represent a major threat to agriculture worldwide. They are transmitted in a persistent, propagative manner by phloem-sucking Hemipteran insects. Phytoplasma membrane proteins are in direct contact with hosts and are presumably involved in determining vector specificity. Such a role has been proposed for phytoplasma transmembrane proteins encoded by circular extrachromosomal elements, at least one of which is a plasmid. Little is known about the interactions between major phytoplasma antigenic membrane protein (Amp and insect vector proteins. The aims of our work were to identify vector proteins interacting with Amp and to investigate their role in transmission specificity. In controlled transmission experiments, four Hemipteran species were identified as vectors of "Candidatus Phytoplasma asteris", the chrysanthemum yellows phytoplasmas (CYP strain, and three others as non-vectors. Interactions between a labelled (recombinant CYP Amp and insect proteins were analysed by far Western blots and affinity chromatography. Amp interacted specifically with a few proteins from vector species only. Among Amp-binding vector proteins, actin and both the α and β subunits of ATP synthase were identified by mass spectrometry and Western blots. Immunofluorescence confocal microscopy and Western blots of plasma membrane and mitochondrial fractions confirmed the localisation of ATP synthase, generally known as a mitochondrial protein, in plasma membranes of midgut and salivary gland cells in the vector Euscelidius variegatus. The vector-specific interaction between phytoplasma Amp and insect ATP synthase is demonstrated for the first time, and this work also supports the hypothesis that host actin is involved in the internalization and intracellular motility of phytoplasmas within their vectors. Phytoplasma Amp is hypothesized to play a crucial role in insect transmission specificity.

  3. Prostate-specific membrane antigen PET/MRI validation of MR textural analysis for detection of transition zone prostate cancer.

    Science.gov (United States)

    Bates, Anthony; Miles, Kenneth

    2017-12-01

    To validate MR textural analysis (MRTA) for detection of transition zone (TZ) prostate cancer through comparison with co-registered prostate-specific membrane antigen (PSMA) PET-MR. Retrospective analysis was performed for 30 men who underwent simultaneous PSMA PET-MR imaging for staging of prostate cancer. Thirty texture features were derived from each manually contoured T2-weighted, transaxial, prostatic TZ using texture analysis software that applies a spatial band-pass filter and quantifies texture through histogram analysis. Texture features of the TZ were compared to PSMA expression on the corresponding PET images. The Benjamini-Hochberg correction controlled the false discovery rate at detection of TZ prostate cancer. • Prostate transition zone (TZ) MR texture analysis may assist in prostate cancer detection. • Abnormal transition zone PSMA expression correlates with altered texture on T2-weighted MR. • TZ with abnormal PSMA expression demonstrates significantly reduced MI, SD and MPP.

  4. Protective effect of intranasal immunization with Neospora caninum membrane antigens against murine neosporosis established through the gastrointestinal tract.

    Science.gov (United States)

    Ferreirinha, Pedro; Dias, Joana; Correia, Alexandra; Pérez-Cabezas, Begoña; Santos, Carlos; Teixeira, Luzia; Ribeiro, Adília; Rocha, António; Vilanova, Manuel

    2014-02-01

    Neospora caninum is an Apicomplexa parasite that in the last two decades was acknowledged as the main pathogenic agent responsible for economic losses in the cattle industry. In the present study, the effectiveness of intranasal immunization with N. caninum membrane antigens plus CpG adjuvant was assessed in a murine model of intragastrically established neosporosis. Immunized mice presented a lower parasitic burden in the brain on infection with 5 × 10(7) tachyzoites, showing that significant protection was achieved by this immunization strategy. Intestinal IgA antibodies raised by immunization markedly agglutinated live N. caninum tachyzoites whereas previous opsonization with IgG antibodies purified from immunized mice sera reduced parasite survival within macrophage cells. Although an IgG1 : IgG2a ratio neosporosis and indicate that parasite-specific mucosal and circulating antibodies have a protective role against this parasitic infection. © 2013 John Wiley & Sons Ltd.

  5. The extracellular membrane-proximal domain of membrane-bound IgE restricts B cell activation by limiting B cell antigen receptor surface expression.

    Science.gov (United States)

    Vanshylla, Kanika; Opazo, Felipe; Gronke, Konrad; Wienands, Jürgen; Engels, Niklas

    2018-03-01

    Immunoglobulin E (IgE) antibodies are key mediators of allergic reactions. Due to their potentially harmful anaphylactic properties, their production is tightly regulated. The membrane-bound isoform of IgE (mIgE), which is an integral component of the B cell antigen receptor, has been shown to be critical for the regulation of IgE responses in mice. In primate species including humans, mIgE can be expressed in two isoforms that are produced by alternative splicing of the primary ε Ig heavy chain transcript, and differ in the absence or presence of an extracellular membrane-proximal domain (EMPD) consisting of 52 amino acids. However, the function of the EMPD remains unclear. Here, we demonstrate that the EMPD restricts surface expression of mIgE-containing BCRs in human and murine B cells. The EMPD does not interfere with BCR assembly but acts as an autonomous endoplasmic reticulum retention domain. Limited surface expression of EMPD-containing mIgE-BCRs caused impaired activation of intracellular signaling cascades and hence represents a regulatory mechanism that may control the production of potentially anaphylactic IgE antibodies in primate species. © 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  6. 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

  7. A radioimmunoassay for antibodies against surface membrane antigens using adhering cells

    International Nuclear Information System (INIS)

    Tax, A.; Manson, L.A.

    1976-01-01

    A radioimmunoassay using cells adhering to plastic is described. In this assay, A-10 mammary carcinoma attached to the surface of plastic in microtiter plates were permitted to bind antibody and the bound antibody was detected with purified rabbit 125 I-antimouse-Fab. The bound radioactive material was eluted with glycine-HCl buffer (pH 2.5), and the acid eluates were counted in a gamma counter. This assay can be used to detect cytolic or noncytolic antibody to cell surface antigens in studies with any tumor or normal cell that will adhere to a solid surface

  8. Prostate-specific membrane antigen (PSMA)-mediated laminin proteolysis generates a pro-angiogenic peptide

    Czech Academy of Sciences Publication Activity Database

    Conway, R. E.; Rojas, C.; Alt, J.; Nováková, Zora; Richardson, S. M.; Rodrick, T. C.; Fuentes, J. L.; Richardson, N. H.; Attalla, J.; Stewart, S.; Fahmy, B.; Bařinka, Cyril; Ghosh, M.; Shapiro, L. H.; Slusher, B. S.

    2016-01-01

    Roč. 19, č. 4 (2016), s. 487-500 ISSN 0969-6970 R&D Projects: GA ČR GAP301/12/1513; GA MŠk(CZ) ED1.1.00/02.0109 Institutional support: RVO:86652036 Keywords : TUMOR-ASSOCIATED NEOVASCULATURE * BASEMENT-MEMBRANE * DISTINCT ANTITUMOR PROPERTIES Subject RIV: FA - Cardiovascular Diseases incl. Cardiotharic Surgery Impact factor: 5.253, year: 2016

  9. Employing Escherichia coli-derived outer membrane vesicles as an antigen delivery platform elicits protective immunity against Acinetobacter baumannii infection

    Science.gov (United States)

    Huang, Weiwei; Wang, Shijie; Yao, Yufeng; Xia, Ye; Yang, Xu; Li, Kui; Sun, Pengyan; Liu, Cunbao; Sun, Wenjia; Bai, Hongmei; Chu, Xiaojie; Li, Yang; Ma, Yanbing

    2016-11-01

    Outer membrane vesicles (OMVs) have proven to be highly immunogenic and induced an immune response against bacterial infection in human clinics and animal models. We sought to investigate whether engineered OMVs can be a feasible antigen-delivery platform for efficiently inducing specific antibody responses. In this study, Omp22 (an outer membrane protein of A. baumannii) was displayed on E. coli DH5α-derived OMVs (Omp22-OMVs) using recombinant gene technology. The morphological features of Omp22-OMVs were similar to those of wild-type OMVs (wtOMVs). Immunization with Omp22-OMVs induced high titers of Omp22-specific antibodies. In a murine sepsis model, Omp22-OMV immunization significantly protected mice from lethal challenge with a clinically isolated A. baumannii strain, which was evidenced by the increased survival rate of the mice, the reduced bacterial burdens in the lung, spleen, liver, kidney, and blood, and the suppressed serum levels of inflammatory cytokines. In vitro opsonophagocytosis assays showed that antiserum collected from Omp22-OMV-immunized mice had bactericidal activity against clinical isolates, which was partly specific antibody-dependent. These results strongly indicated that engineered OMVs could display a whole heterologous protein (~22 kDa) on the surface and effectively induce specific antibody responses, and thus OMVs have the potential to be a feasible vaccine platform.

  10. Protective effect of intranasal immunization with Neospora caninum membrane antigens against murine neosporosis established through the gastrointestinal tract

    Science.gov (United States)

    Ferreirinha, Pedro; Dias, Joana; Correia, Alexandra; Pérez-Cabezas, Begoña; Santos, Carlos; Teixeira, Luzia; Ribeiro, Adília; Rocha, António; Vilanova, Manuel

    2014-01-01

    Neospora caninum is an Apicomplexa parasite that in the last two decades was acknowledged as the main pathogenic agent responsible for economic losses in the cattle industry. In the present study, the effectiveness of intranasal immunization with N. caninum membrane antigens plus CpG adjuvant was assessed in a murine model of intragastrically established neosporosis. Immunized mice presented a lower parasitic burden in the brain on infection with 5 × 107 tachyzoites, showing that significant protection was achieved by this immunization strategy. Intestinal IgA antibodies raised by immunization markedly agglutinated live N. caninum tachyzoites whereas previous opsonization with IgG antibodies purified from immunized mice sera reduced parasite survival within macrophage cells. Although an IgG1 : IgG2a ratio < 1 was detected in the immunized mice before and after infection, indicative of a predominant T helper type 1 immune response, no increased production of interferon-γ was detected in the spleen or mesenteric lymph nodes of the immunized mice. Altogether, these results show that mucosal immunization with N. caninum membrane proteins plus CpG adjuvant protect against intragastrically established neosporosis and indicate that parasite-specific mucosal and circulating antibodies have a protective role against this parasitic infection. PMID:24128071

  11. 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

    The F-antigen is a prominent liver protein which has been extensively used in studies on natural and induced immunological tolerance. However, its intracellular localization and biological function have remained elusive. It has generally been assumed that the F-antigen is confined phylogenetically...... of the TF-ag protein, however, declined only moderately during prolonged periods of starvation demonstrating that extensive release of the TF-ag did not take place. In combination these results suggest that the TF-ag protein is a recycled constituent of the intracellular membrane network in T. thermophila...... to vertebrates. Now we have cloned and characterized a gene from the ciliated protozoan Tetrahymena thermophila encoding a protein which clearly is homologous with the rat F-antigen. The coding region of the Tetrahymena F-antigen (TF-ag) gene specifies a 46,051 M(r) protein and is interrupted by three introns...

  12. Different immunogenicity but similar antitumor efficacy of two DNA vaccines coding for an antigen secreted in different membrane vesicle-associated forms

    Science.gov (United States)

    Sedlik, Christine; Vigneron, James; Torrieri-Dramard, Lea; Pitoiset, Fabien; Denizeau, Jordan; Chesneau, Caroline; de la Rochere, Philippe; Lantz, Olivier; Thery, Clotilde; Bellier, Bertrand

    2014-01-01

    The induction of an active immune response to control or eliminate tumours is still an unfulfilled challenge. We focused on plasmid DNA vaccines using an innovative approach whereby the antigen is expressed in association with extracellular vesicles (EVs) to facilitate antigen cross-presentation and improve induced immunity. Our two groups had independently shown previously that DNA vaccines encoding EV-associated antigens are more efficient at inducing cytotoxic T-cell responses than vaccines encoding the non-EV-associated antigen. Here, we compared our two approaches to associate the ovalbumin (OVA) antigen to EVs: (a) by fusion to the lipid-binding domain C1C2 of MFGE8(=lactadherin), which is exposed on the surface of secreted membrane vesicles; and (b) by fusion to retroviral Gag capsid protein, which is incorporated inside membrane-enclosed virus-like particles. Plasmids encoding either form of modified OVA were used as DNA-based vaccines (i.e. injected into mice to allow in vivo expression of the antigen associated to EVs). We show that both DNA vaccines induced, with similar efficiency, OVA-specific CD8+ T cells and total IgG antibodies. By contrast, each vaccine preferentially stimulated different isotypes of immunoglobulins, and the OVA-C1C2-encoding vaccine favoured antigen-specific CD4+ T lymphocyte induction as compared to the Gag-OVA vaccine. Nevertheless, both OVA-C1C2 and Gag-OVA vaccines efficiently prevented in vivo outgrowth of OVA-expressing tumours and reduced tumour progression when administered to tumour-bearing mice, although with variable efficacies depending on the tumour models. DNA vaccines encoding EV-associated antigens are thus promising immunotherapy tools in cancer but also potentially other diseases. PMID:25206960

  13. Different immunogenicity but similar antitumor efficacy of two DNA vaccines coding for an antigen secreted in different membrane vesicle-associated forms

    Directory of Open Access Journals (Sweden)

    Christine Sedlik

    2014-08-01

    Full Text Available The induction of an active immune response to control or eliminate tumours is still an unfulfilled challenge. We focused on plasmid DNA vaccines using an innovative approach whereby the antigen is expressed in association with extracellular vesicles (EVs to facilitate antigen cross-presentation and improve induced immunity. Our two groups had independently shown previously that DNA vaccines encoding EV-associated antigens are more efficient at inducing cytotoxic T-cell responses than vaccines encoding the non-EV-associated antigen. Here, we compared our two approaches to associate the ovalbumin (OVA antigen to EVs: (a by fusion to the lipid-binding domain C1C2 of MFGE8(=lactadherin, which is exposed on the surface of secreted membrane vesicles; and (b by fusion to retroviral Gag capsid protein, which is incorporated inside membrane-enclosed virus-like particles. Plasmids encoding either form of modified OVA were used as DNA-based vaccines (i.e. injected into mice to allow in vivo expression of the antigen associated to EVs. We show that both DNA vaccines induced, with similar efficiency, OVA-specific CD8+ T cells and total IgG antibodies. By contrast, each vaccine preferentially stimulated different isotypes of immunoglobulins, and the OVA-C1C2-encoding vaccine favoured antigen-specific CD4+ T lymphocyte induction as compared to the Gag-OVA vaccine. Nevertheless, both OVA-C1C2 and Gag-OVA vaccines efficiently prevented in vivo outgrowth of OVA-expressing tumours and reduced tumour progression when administered to tumour-bearing mice, although with variable efficacies depending on the tumour models. DNA vaccines encoding EV-associated antigens are thus promising immunotherapy tools in cancer but also potentially other diseases.

  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

    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

  15. Formulation of an inhibitor radiopharmaceutical of prostatic antigen of 177Lu-Glu-Nh-CO-Nh-Lys membrane

    International Nuclear Information System (INIS)

    Ortega S, D.

    2015-01-01

    The prostate specific membrane antigen (PSMA) is a zinc metalloenzyme that is expressed on the cell membrane and highly expressed in prostate cancer. Recently, it has been demonstrated that the peptide sequence Glu-Nh-CO-Nh-Lys inhibit PSMA activity through an electrostatic interaction with the Zn. Several theragnostic radiopharmaceuticals with base in 177 Lu have been developed for radiotherapy of specific molecular targets because gamma and beta emissions of the radionuclide (β = 0.498 MeV and γ= 0.133 MeV). However, there is currently no label a formulation for preparing a radiopharmaceutical of 177 Lu-Glu-Nh-CO-Nh-Lys useful treatment of prostate cancer. The aim of this research was to optimize and document the process of production of the radiopharmaceutical 177 Lu-Glu-Nh-CO-Nh-Lys for sanitary registration application before the Comision Federal para la Proteccion contra Riesgos Sanitarios (COFEPRIS). The optimization of the production process was assessed a factorial design of three variables with mixed levels (3 x 3 x 2) where the dependent variable is the radiochemical purity, the analytical method was validated by UV-Vis spectrophotometry. Next, process validation was carried out by labeling 3 lots of the optimized formulation of the radiopharmaceutical (5.55 GBq (2.16 μg) of 177 LuCl 3 , 90 mg peptide PSMA, 50 mg ascorbic acid and 150 μL of acetate buffer 1 M ph 5), long-term stability was performed by high resolution liquid chromatography) to determine its useful shelf life. 3 validation batches were prepared under protocols of Good Manufacturing Practice (GMP) in the Production Plant of Radiopharmaceuticals of the Instituto Nacional de Investigaciones Nucleares (ININ), meet specifications preset by obtaining a sterile and free development of bacterial endotoxin yields of labeled 100% and which retains its quality characteristics radiochemical purity greater than 90% for at least 15 days. (Author)

  16. Aminopeptidase N is directly sorted to the apical domain in MDCK cells

    DEFF Research Database (Denmark)

    Wessels, H P; Hansen, Gert Helge; Fuhrer, C

    1990-01-01

    In different epithelial cell types, integral membrane proteins appear to follow different sorting pathways to the apical surface. In hepatocytes, several apical proteins were shown to be transported there indirectly via the basolateral membrane, whereas in MDCK cells a direct sorting pathway from...

  17. Pearls and pitfalls in clinical interpretation of prostate-specific membrane antigen (PSMA)-targeted PET imaging

    International Nuclear Information System (INIS)

    Sheikhbahaei, Sara; Solnes, Lilja B.; Javadi, Mehrbod S.; Pomper, Martin G.; Rowe, Steven P.; Afshar-Oromieh, Ali; Haberkorn, Uwe; Eiber, Matthias; Ross, Ashley E.; Pienta, Kenneth J.; Allaf, Mohamad E.; Gorin, Michael A.

    2017-01-01

    The rapidly expanding clinical adaptation of prostate-specific membrane antigen (PSMA)-targeted PET imaging in the evaluation of patients with prostate cancer has placed an increasing onus on understanding both the potential pearls of interpretation as well as limitations of this new technique. As with any new molecular imaging modality, accurate characterization of abnormalities on PSMA-targeted PET imaging can be accomplished only if one is aware of the normal distribution pattern, physiological variants of radiotracer uptake, and potential sources of false-positive and false-negative imaging findings. In recent years, a growing number of reports have come to light describing incidental non-prostatic benign or malignant pathologies with high uptake on PSMA-targeted PET imaging. In this review, we have summarized the published literature regarding the potential pearls and technical and interpretive pitfalls of this imaging modality. Knowledge of these limitations can increase the confidence of interpreting physicians and thus improve patient care. As PSMA-targeted PET is expected to be evaluated in larger prospective trials, the dissemination of potential diagnostic pitfalls and the biologic underpinning of those findings will be of increased importance. (orig.)

  18. Pearls and pitfalls in clinical interpretation of prostate-specific membrane antigen (PSMA)-targeted PET imaging

    Energy Technology Data Exchange (ETDEWEB)

    Sheikhbahaei, Sara; Solnes, Lilja B.; Javadi, Mehrbod S.; Pomper, Martin G.; Rowe, Steven P. [Johns Hopkins University School of Medicine, Division of Nuclear Medicine and Molecular Imaging, The Russell H. Morgan Department of Radiology and Radiological Science, Baltimore, MD (United States); Afshar-Oromieh, Ali; Haberkorn, Uwe [Heidelberg University Hospital, Department of Nuclear Medicine, Heidelberg (Germany); Eiber, Matthias [David Geffen School of Medicine at UCLA, Department of Molecular and Medical Pharmacology, Los Angeles, CA (United States); Technical University of Munich, Department of Nuclear Medicine, Klinikum rechts der Isar, Munich (Germany); Ross, Ashley E.; Pienta, Kenneth J.; Allaf, Mohamad E.; Gorin, Michael A. [Johns Hopkins University School of Medicine, The James Buchanan Brady Urological Institute and Department of Urology, Baltimore, MD (United States)

    2017-11-15

    The rapidly expanding clinical adaptation of prostate-specific membrane antigen (PSMA)-targeted PET imaging in the evaluation of patients with prostate cancer has placed an increasing onus on understanding both the potential pearls of interpretation as well as limitations of this new technique. As with any new molecular imaging modality, accurate characterization of abnormalities on PSMA-targeted PET imaging can be accomplished only if one is aware of the normal distribution pattern, physiological variants of radiotracer uptake, and potential sources of false-positive and false-negative imaging findings. In recent years, a growing number of reports have come to light describing incidental non-prostatic benign or malignant pathologies with high uptake on PSMA-targeted PET imaging. In this review, we have summarized the published literature regarding the potential pearls and technical and interpretive pitfalls of this imaging modality. Knowledge of these limitations can increase the confidence of interpreting physicians and thus improve patient care. As PSMA-targeted PET is expected to be evaluated in larger prospective trials, the dissemination of potential diagnostic pitfalls and the biologic underpinning of those findings will be of increased importance. (orig.)

  19. Prostate-specific membrane antigen PET/MRI validation of MR textural analysis for detection of transition zone prostate cancer

    Energy Technology Data Exchange (ETDEWEB)

    Bates, Anthony [Princess Alexandra Hospital, Brisbane (Australia); Miles, Kenneth [Princess Alexandra Hospital, Department of Diagnostic Radiology, Brisbane, QLD (Australia); University College London, Institute of Nuclear Medicine, London (United Kingdom)

    2017-12-15

    To validate MR textural analysis (MRTA) for detection of transition zone (TZ) prostate cancer through comparison with co-registered prostate-specific membrane antigen (PSMA) PET-MR. Retrospective analysis was performed for 30 men who underwent simultaneous PSMA PET-MR imaging for staging of prostate cancer. Thirty texture features were derived from each manually contoured T2-weighted, transaxial, prostatic TZ using texture analysis software that applies a spatial band-pass filter and quantifies texture through histogram analysis. Texture features of the TZ were compared to PSMA expression on the corresponding PET images. The Benjamini-Hochberg correction controlled the false discovery rate at <5%. Eighty-eight T2-weighted images in 18 patients demonstrated abnormal PSMA expression within the TZ on PET-MR. 123 images were PSMA negative. Based on the corrected p-value of 0.005, significant differences between PSMA positive and negative slices were found for 16 texture parameters: Standard deviation and mean of positive pixels for all spatial filters (p = <0.0001 for both at all spatial scaling factor (SSF) values) and mean intensity following filtration for SSF 3-6 mm (p = 0.0002-0.0018). Abnormal expression of PSMA within the TZ is associated with altered texture on T2-weighted MR, providing validation of MRTA for the detection of TZ prostate cancer. (orig.)

  20. Unusual Self-Assembly of the Recombinant Chlamydia trachomatis Major Outer Membrane Protein-Based Fusion Antigen CTH522 Into Protein Nanoparticles

    DEFF Research Database (Denmark)

    Rose, Fabrice; Karlsen, Kasper; Jensen, Pernille

    2018-01-01

    but is a challenging vaccine candidate by being an integral membrane protein, and the immunogenicity depends on a correctly folded structure. We investigated the biophysical properties of the recombinant MOMP-based fusion antigen CTH522, which is tested in early human clinical trials. It consists of a truncated......Sexually transmitted Chlamydia trachomatis (Ct) infects more than 100 million people annually, and untreated chlamydia infections can cause severe complications. Therefore, there is an urgent need for a chlamydia vaccine. The Ct major outer membrane protein (MOMP) is highly immunogenic...

  1. Prostate cancer nodal oligometastasis accurately assessed using prostate-specific membrane antigen positron emission tomography-computed tomography and confirmed histologically following robotic-assisted lymph node dissection

    Directory of Open Access Journals (Sweden)

    Dermot B O′Kane

    2016-01-01

    Full Text Available We herein present a case of a 76-year-old gentleman, where prostate-specific membrane antigen positron emission tomography-computed tomography (PSMA PET-CT was used to accurately detect prostate cancer (PCa, pelvic lymph node (LN metastasis in the setting of biochemical recurrence following definitive treatment for PCa. The positive PSMA PET-CT result was confirmed with histological examination of the involved pelvic LNs following pelvic LN dissection.

  2. Prostate cancer nodal oligometastasis accurately assessed using prostate-specific membrane antigen positron emission tomography-computed tomography and confirmed histologically following robotic-assisted lymph node dissection.

    Science.gov (United States)

    O'Kane, Dermot B; Lawrentschuk, Nathan; Bolton, Damien M

    2016-01-01

    We herein present a case of a 76-year-old gentleman, where prostate-specific membrane antigen positron emission tomography-computed tomography (PSMA PET-CT) was used to accurately detect prostate cancer (PCa), pelvic lymph node (LN) metastasis in the setting of biochemical recurrence following definitive treatment for PCa. The positive PSMA PET-CT result was confirmed with histological examination of the involved pelvic LNs following pelvic LN dissection.

  3. Vascular targeted therapy with anti-prostate-specific membrane antigen monoclonal antibody J591 in advanced solid tumors.

    Science.gov (United States)

    Milowsky, Matthew I; Nanus, David M; Kostakoglu, Lale; Sheehan, Christine E; Vallabhajosula, Shankar; Goldsmith, Stanley J; Ross, Jeffrey S; Bander, Neil H

    2007-02-10

    Based on prostate-specific membrane antigen (PSMA) expression on the vasculature of solid tumors, we performed a phase I trial of antibody J591, targeting the extracellular domain of PSMA, in patients with advanced solid tumor malignancies. This was a proof-of-principle evaluation of PSMA as a potential neovascular target. The primary end points were targeting,toxicity, maximum-tolerated dose, pharmacokinetics (PK), and human antihuman antibody (HAHA) response. Patients had advanced solid tumors previously shown to express PSMA on the neovasculature. They received 111Indium (111ln)-J591 for scintigraphy and PK, followed 2 weeks later by J591 with a reduced amount of 111In for additional PK measurements. J591 dose levels were 5, 10, 20, 40, and 80 mg. The protocol was amended for six weekly administrations of unchelated J591. Patients with a response or stable disease were eligible for re-treatment. Immunohistochemistry assessed PSMA expression in tumor tissues. Twenty-seven patients received monoclonal antibody (mAb) J591. Treatment was well tolerated. Twenty (74%) of 27 patients had at least one area of known metastatic disease targeted by 111In-J591, with positive imaging seen in patients with kidney, bladder, lung, breast, colorectal, and pancreatic cancers, and melanoma. Seven of 10 patient specimens available for immunohistochemical assessment of PSMA expression in tumor-associated vasculature demonstrated PSMA staining. No HAHA response was seen. Three patients of 27 with stable disease received re-treatment. Acceptable toxicity and excellent targeting of known sites of metastases were demonstrated in patients with multiple solid tumor types, highlighting a potential role for the anti-PSMA antibody J591 as a vascular-targeting agent.

  4. 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

    2016-11-01

    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.

  5. Abceso apical agudo

    OpenAIRE

    Jara Ortiz, M.; López, M.; Gómez Viglino, L.; Maydana, N.; Hervit, M.; Bertola, N.

    2015-01-01

    En casos de urgencia, muchas veces debemos poner a prueba distintos elementos de diagnóstico para encontrar la mejor respuesta posible. En este caso se reporta un absceso apical agudo en la zona palatina del sector 1, que, de acuerdo al test de vitalidad pulpar y correcta interpretación radiográfica se localiza la pieza causal N° 1.8.Asistió a la consulta un paciente masculino 30 años de edad, dolor a la masticación, edema y tumefacción, en zona palatina comprendida en piezas 1.5; 1.6; 1.7 y ...

  6. Apical trafficking in epithelial cells: signals, clusters and motors.

    Science.gov (United States)

    Weisz, Ora A; Rodriguez-Boulan, Enrique

    2009-12-01

    In the early days of epithelial cell biology, researchers working with kidney and/or intestinal epithelial cell lines and with hepatocytes described the biosynthetic and recycling routes followed by apical and basolateral plasma membrane (PM) proteins. They identified the trans-Golgi network and recycling endosomes as the compartments that carried out apical-basolateral sorting. They described complex apical sorting signals that promoted association with lipid rafts, and simpler basolateral sorting signals resembling clathrin-coated-pit endocytic motifs. They also noticed that different epithelial cell types routed their apical PM proteins very differently, using either a vectorial (direct) route or a transcytotic (indirect) route. Although these original observations have generally held up, recent studies have revealed interesting complexities in the routes taken by apically destined proteins and have extended our understanding of the machinery required to sustain these elaborate sorting pathways. Here, we critically review the current status of apical trafficking mechanisms and discuss a model in which clustering is required to recruit apical trafficking machineries. Uncovering the mechanisms responsible for polarized trafficking and their epithelial-specific variations will help understand how epithelial functional diversity is generated and the pathogenesis of many human diseases.

  7. Development of a Ga-68 labeled PET tracer with short linker for prostate-specific membrane antigen (PSMA) targeting.

    Science.gov (United States)

    Moon, Sung-Hyun; Hong, Mee Kyung; Kim, Young Ju; Lee, Yun-Sang; Lee, Dong Soo; Chung, June-Key; Jeong, Jae Min

    2018-04-05

    Glu-Urea-Lys (GUL) derivatives have been reported as prostate-specific membrane antigen (PSMA) agent. We developed derivatives of GUL conjugated with NOTA or DOTA via a thiourea linker and tested their feasibility as PSMA imaging agents after labeling with 68 Ga. NOTA-GUL and DOTA-GUL were synthesized and labeled with 68 Ga using generator-eluted 68 GaCl 3 in 0.1 M HCl in the presence of 1 M NaOAc at pH 5.5. The stabilities of 68 Ga-labeled compounds in human serum were tested at 37.5 °C. A competitive binding assay was performed using the PSMA-positive prostate cancer cell line 22Rv1 and [ 125 I]MIP-1072 (PSMA-specific binding agent) as a tracer. Biodistribution and micro-PET studies were performed using 22Rv1-xenograft BALB/c nude mice. The radiolabeling efficiency of NOTA-GUL (>99%) was higher than that of DOTA-GUL (92%). The IC 50 of Ga-NOTA-GUL was 18.3 nM. In the biodistribution study, tumor uptake of 68 Ga-NOTA-GUL (5.40% ID/g) was higher than that of 68 Ga-DOTA-GUL (4.66% ID/g) at 1 h. Tumor/muscle and tumor/blood uptake ratios of 68 Ga-NOTA-GUL (31.8 and 135, respectively) were significantly higher than those of 68 Ga-DOTA-GUL (16.1 and 31.1, respectively). The tumor/kidney uptake ratio of 68 Ga-NOTA-GUL was 3.4-fold higher than that of 68 Ga-DOTA-GUL. 68 Ga-NOTA-GUL showed specific uptake to PSMA positive tumor xenograft and was blocked by co-injection of the cold ligand. In conclusion, we successfully synthesized 68 Ga-NOTA-GUL and 68 Ga-DOTA-GUL for prostate cancer imaging. 68 Ga-NOTA-GUL showed better radiochemical and biodistribution results. 68 Ga-NOTA-GUL may be a promising PSMA targeting radiopharmaceutical. Copyright © 2018 Elsevier Ltd. All rights reserved.

  8. Recombinant major outer membrane protein (MOMP) of Chlamydophila abortus, Chlamydophila pecorum, and Chlamydia suis as antigens to distinguish chlamydial species-specific antibodies in animal sera.

    Science.gov (United States)

    Hoelzle, Ludwig E; Hoelzle, Katharina; Wittenbrink, Max M

    2004-10-05

    Recombinant major outer membrane proteins (rMOMP) of Chlamydophila (Ch.) abortus, Ch. pecorum, and Chlamydia (C.) suis were used as antigens to distinguish chlamydial species-specific antibodies in (i) immune sera from six rabbits and three pigs raised against native purified elementary bodies, (ii) serum samples from 25 sows vaccinated with Ch. abortus, and (iii) 40 serum samples from four heifers experimentally infected with Ch. abortus. All post-exposition sera contained chlamydial antibodies as confirmed by strong ELISA seroreactivities against the chlamydial LPS. For the rMOMP ELISA mean IgG antibody levels were at least 5.8-fold higher with the particular rMOMP homologous to the chlamydial species used for immunisation or infection than with heterologous rMOMPs (P <0.001). Preferential rMOMP ELISA reactivities of sera were confirmed by Western blotting. The results suggest that the entire chlamydial rMOMP could provide a species-specific serodiagnostic antigen.

  9. Unusual Self-Assembly of the Recombinant Chlamydia trachomatis Major Outer Membrane Protein-Based Fusion Antigen CTH522 Into Protein Nanoparticles

    DEFF Research Database (Denmark)

    Rose, Fabrice; Karlsen, Kasper; Jensen, Pernille

    2018-01-01

    but is a challenging vaccine candidate by being an integral membrane protein, and the immunogenicity depends on a correctly folded structure. We investigated the biophysical properties of the recombinant MOMP-based fusion antigen CTH522, which is tested in early human clinical trials. It consists of a truncated......Sexually transmitted Chlamydia trachomatis (Ct) infects more than 100 million people annually, and untreated chlamydia infections can cause severe complications. Therefore, there is an urgent need for a chlamydia vaccine. The Ct major outer membrane protein (MOMP) is highly immunogenic......-defined secondary structural elements, and no thermal transitions were measurable. Chemical unfolding resulted monomers that upon removal of the denaturant self-assembled into higher order structures, comparable to the structure of the native protein. The conformation of CTH522 in nanoparticles is thus not entirely...

  10. Modulation of endotoxicity of Shigella generalized modules for membrane antigens (GMMA) by genetic lipid A modifications: relative activation of TLR4 and TLR2 pathways in different mutants.

    Science.gov (United States)

    Rossi, Omar; Pesce, Isabella; Giannelli, Carlo; Aprea, Susanna; Caboni, Mariaelena; Citiulo, Francesco; Valentini, Sara; Ferlenghi, Ilaria; MacLennan, Calman Alexander; D'Oro, Ugo; Saul, Allan; Gerke, Christiane

    2014-09-05

    Outer membrane particles from Gram-negative bacteria are attractive vaccine candidates as they present surface antigens in their natural context. We previously developed a high yield production process for genetically derived particles, called generalized modules for membrane antigens (GMMA), from Shigella. As GMMA are derived from the outer membrane, they contain immunostimulatory components, especially lipopolysaccharide (LPS). We examined ways of reducing their reactogenicity by modifying lipid A, the endotoxic part of LPS, through deletion of late acyltransferase genes, msbB or htrB, in GMMA-producing Shigella sonnei and Shigella flexneri strains. GMMA with resulting penta-acylated lipid A from the msbB mutants showed a 600-fold reduced ability, and GMMA from the S. sonnei ΔhtrB mutant showed a 60,000-fold reduced ability compared with GMMA with wild-type lipid A to stimulate human Toll-like receptor 4 (TLR4) in a reporter cell line. In human peripheral blood mononuclear cells, GMMA with penta-acylated lipid A showed a marked reduction in induction of inflammatory cytokines (S. sonnei ΔhtrB, 800-fold; ΔmsbB mutants, 300-fold). We found that the residual activity of these GMMA is largely due to non-lipid A-related TLR2 activation. In contrast, in the S. flexneri ΔhtrB mutant, a compensatory lipid A palmitoleoylation resulted in GMMA with hexa-acylated lipid A with ∼10-fold higher activity to stimulate peripheral blood mononuclear cells than GMMA with penta-acylated lipid A, mostly due to retained TLR4 activity. Thus, for use as vaccines, GMMA will likely require lipid A penta-acylation. The results identify the relative contributions of TLR4 and TLR2 activation by GMMA, which need to be taken into consideration for GMMA vaccine development. © 2014 by The American Society for Biochemistry and Molecular Biology, Inc.

  11. Evaluation of Protective Potential of Yersinia pestis Outer Membrane Protein Antigens as Possible Candidates for a New-Generation Recombinant Plague Vaccine

    Science.gov (United States)

    Erova, Tatiana E.; Rosenzweig, Jason A.; Sha, Jian; Suarez, Giovanni; Sierra, Johanna C.; Kirtley, Michelle L.; van Lier, Christina J.; Telepnev, Maxim V.; Motin, Vladimir L.

    2013-01-01

    Plague caused by Yersinia pestis manifests itself in bubonic, septicemic, and pneumonic forms. Although the U.S. Food and Drug Administration recently approved levofloxacin, there is no approved human vaccine against plague. The capsular antigen F1 and the low-calcium-response V antigen (LcrV) of Y. pestis represent excellent vaccine candidates; however, the inability of the immune responses to F1 and LcrV to provide protection against Y. pestis F1− strains or those which harbor variants of LcrV is a significant concern. Here, we show that the passive transfer of hyperimmune sera from rats infected with the plague bacterium and rescued by levofloxacin protected naive animals against pneumonic plague. Furthermore, 10 to 12 protein bands from wild-type (WT) Y. pestis CO92 reacted with the aforementioned hyperimmune sera upon Western blot analysis. Based on mass spectrometric analysis, four of these proteins were identified as attachment invasion locus (Ail/OmpX), plasminogen-activating protease (Pla), outer membrane protein A (OmpA), and F1. The genes encoding these proteins were cloned, and the recombinant proteins purified from Escherichia coli for immunization purposes before challenging mice and rats with either the F1− mutant or WT CO92 in bubonic and pneumonic plague models. Although antibodies to Ail and OmpA protected mice against bubonic plague when challenged with the F1− CO92 strain, Pla antibodies were protective against pneumonic plague. In the rat model, antibodies to Ail provided protection only against pneumonic plague after WT CO92 challenge. Together, the addition of Y. pestis outer membrane proteins to a new-generation recombinant vaccine could provide protection against a wide variety of Y. pestis strains. PMID:23239803

  12. The use of outer membrane proteins as an exposure surface for foreign antigens in AIDS vaccine methodology and AIDS diagnostics

    NARCIS (Netherlands)

    Soede WWD; Hegger I

    1992-01-01

    A live recombinant bacteria or virus with HIV determinants exposed at the outermembrane is one strategy for AIDS vaccine development. Two HIV determinants that showed neutralization capacity in in- vitro experiments were tested for their expression in PhoE outer membrane protein of E coli K12.

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

    Directory of Open Access Journals (Sweden)

    Damer P Blake

    2011-02-01

    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

  14. Sensitivity, Specificity, and Predictors of Positive68Ga-Prostate-specific Membrane Antigen Positron Emission Tomography in Advanced Prostate Cancer: A Systematic Review and Meta-analysis.

    Science.gov (United States)

    Perera, Marlon; Papa, Nathan; Christidis, Daniel; Wetherell, David; Hofman, Michael S; Murphy, Declan G; Bolton, Damien; Lawrentschuk, Nathan

    2016-12-01

    Positron emission tomography (PET) of 68 Ga-labelled prostate-specific membrane antigen ( 68 Ga-PSMA) is an emerging imaging modality introduced to assess the burden of prostate cancer, typically in biochemically recurrent or advanced disease. 68 Ga-PSMA PET provides the ability to selectively identify and localize metastatic prostate cancer cells and subsequently change patient management. Owing to its limited history, robust sensitivity and specificity data are not available for 68 Ga-PSMA PET-positive scans. A systematic review and meta-analysis of reported predictors of positive 68 Ga-PSMA PET and corresponding sensitivity and specificity profiles. We performed critical reviews of MEDLINE, EMBASE, ScienceDirect, Cochrane Library, and Web of Science databases in April 2016 according to the Preferred Reporting Items for Systematic Review and Meta-analysis (PRISMA) statement. Quality was assessed using the Quality Assessment if Diagnostic Accuracy Studies-2 tool. Meta-analysis and meta-regression of proportions were performed using a random-effects model with pre-PET prostate-specific antigen (PSA) levels as the dependent variable. Summary sensitivity and specificity values were obtained by fitting bivariate hierarchical regression models. Sixteen articles involving 1309 patients were analysed. The overall percentage of positive 68 Ga-PSMA PET among patients was 40% (95% confidence interval [CI] 19-64%) for primary staging and 76% (95% CI 66-85%) for biochemical recurrence (BCR). Positive 68 Ga-PSMA PET scans for BCR patients increased with pre-PET PSA. For the PSA categories 0-0.2, 0.2-1, 1-2, and >2 ng/ml, 42%, 58%, 76%, and 95% scans, respectively, were positive. Shorter PSA doubling time increased 68 Ga-PSMA PET positivity. On per-patient analysis, the summary sensitivity and specificity were both 86%. On per-lesion analysis, the summary sensitivity and specificity were 80% and 97%, respectively. In the setting of BCR prostate cancer, pre-PET PSA predicts the

  15. 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.

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

    International Nuclear Information System (INIS)

    Kalra, Rajkumar S.; Wadhwa, Renu

    2015-01-01

    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 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

  17. Comparison of clinical performance of antigen basedenzyme immunoassay (EIA and major outer membrane protein (MOMP-PCR for detection of genital Chlamydia trachomatis infection

    Directory of Open Access Journals (Sweden)

    Mahmoud Nateghi Rostami

    2016-06-01

    Full Text Available Background: Chlamydia trachomatis is the most common sexually transmitted bacterial pathogen worldwide. Early detection and treatment of C.trachomatis genital infection prevent serious reproductive complications. Objective: Performances of enzyme immunoassay (EIA and major outer membrane protein (MOMP-polymerase chain reaction (PCR for diagnosis of genital C.trachomatis infection in women were compared. Materials and Methods: In this cross sectional study a total of 518 women volunteers were included (33.67±8.3 yrs who had been referred to Gynecology clinics of Qom province, Iran, were included. Endocervical swab specimens were collected to detect lipopolysaccharide (LPS antigen in EIA and to amplify MOMP gene of C.trachomatis in PCR. Results were confirmed using ompI nested-PCR. Sensitivity, specificity, positive (PPV and negative predictive values (NPV were calculated for performance of the tests. Odds ratios were determined using binary logistic regression analysis. Results: In total, 37 (7.14% cases were positive by EIA and/or MOMP-PCR. All discrepant results were confirmed by nested-PCR. Sensitivity, specificity, PPV and NPV values of EIA were 59.46%, 100%, 100% and 96.98%, and those of MOMPPCR were 97.30%, 100%, 100%, 99.79%, respectively. Reproductive complications including 2.7% ectopic pregnancy, 5.4% stillbirth, 5.4% infertility, and 10.8% PROM were recorded. The risk of developing chlamydiosis was increased 4.8-fold in volunteers with cervicitis (p<0.05; OR 4.80; 95% CI 1.25-18.48. Conclusion: C.trachomatis infection should be regarded in women of reproductive ages especially those with cervicitis. Primary screening of women by using the low cost antigen-EIA is recommended; however, due to the low sensitivity of Ag-EIA, verification of the negative results by a DNA amplification method is needed.

  18. Phase I trial of yttrium-90-labeled anti-prostate-specific membrane antigen monoclonal antibody J591 for androgen-independent prostate cancer.

    Science.gov (United States)

    Milowsky, Matthew I; Nanus, David M; Kostakoglu, Lale; Vallabhajosula, Shankar; Goldsmith, Stanley J; Bander, Neil H

    2004-07-01

    To determine the maximum-tolerated dose (MTD), toxicity, human antihuman antibody (HAHA) response, pharmacokinetics, organ dosimetry, targeting, and preliminary efficacy of yttrium-90-labeled anti-prostate-specific membrane antigen monoclonal antibody J591 ((90)Y-J591) in patients with androgen-independent prostate cancer (PC). Patients with androgen-independent PC and evidence of disease progression received indium-111-J591 for pharmacokinetic and biodistribution determinations followed 1 week later by (90)Y-J591 at five dose levels: 5, 10, 15, 17.5, and 20 mCi/m(2). Patients were eligible for up to three re-treatments if platelet and neutrophil recovery was satisfactory. Twenty-nine patients with androgen-independent PC received (90)Y-J591, four of whom were re-treated. Dose limiting toxicity (DLT) was seen at 20 mCi/m(2), with two patients experiencing thrombocytopenia with non-life-threatening bleeding episodes requiring platelet transfusions. The 17.5-mCi/m(2) dose level was determined to be the MTD. No re-treated patients experienced DLT. Nonhematologic toxicity was not dose limiting. Targeting of known sites of bone and soft tissue metastases was seen in the majority of patients. No HAHA response was seen. Antitumor activity was seen, with two patients experiencing 85% and 70% declines in prostate-specific antigen (PSA) levels lasting 8 and 8.6 months, respectively, before returning to baseline. Both patients had objective measurable disease responses. An additional six patients (21%) experienced PSA stabilization. The recommended dose for (90)Y-J591 is 17.5 mCi/m(2). Acceptable toxicity, excellent targeting of known sites of PC metastases, and biologic activity in patients with androgen-independent PC warrant further investigation of (90)Y-J591 in the treatment of patients with PC.

  19. 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. PMID:27180806

  20. Left ventricular apical ballooning syndrome

    International Nuclear Information System (INIS)

    Rahman, N.; Tai, J.; Soofi, A.

    2007-01-01

    The transient left ventricular apical ballooning syndrome, also known as Takotsubo cardiomyopathy, is characterized by transient left ventricular dysfunction in the absence of obstructive epicardial coronary disease. Although the syndrome has been reported in Japan since 1990, it is rare in other regions. Rapid recognition of the syndrome can modify the diagnostic and therapeutic attitude i.e. avoiding thrombolysis and performing catheterization in the acute phase. (author)

  1. Comparison of Intranasal Outer Membrane Vesicles with Cholera Toxin and Injected MF59C.1 as Adjuvants for Malaria Transmission Blocking Antigens AnAPN1 and Pfs48/45

    Directory of Open Access Journals (Sweden)

    Michael Pritsch

    2016-01-01

    Full Text Available Purified protein vaccines often require adjuvants for efficient stimulation of immune responses. There is no licensed mucosal adjuvant on the market to adequately boost the immune response to purified antigens for intranasal applications in humans. Bacterial outer membrane vesicles (OMV are attractive candidates potentially combining antigenic and adjuvant properties in one substance. To more precisely characterize the potential of Escherichia coli OMV for intranasal vaccination with heterologous antigens, immune responses for AnAPN1 and Pfs48/45 as well as ovalbumin as a reference antigen were assessed in mice. The intranasal adjuvant cholera toxin (CT and parenteral adjuvant MF59C.1 were used in comparison. Vaccinations were administered intranasally or subcutaneously. Antibodies (total IgG and IgM as well as subclasses IgG1, IgG2a, IgG2b, and IgG3 were measured by ELISA. T cell responses (cytotoxic T cells, Th1, Th17, and regulatory T cells were determined by flow cytometry. When OMV were used as adjuvant for intranasal immunization, antibody and cellular responses against all three antigens could be induced, comparable to cholera toxin and MF59C.1. Antigen-specific IgG titres above 1 : 105 could be detected in all groups. This study provides the rationale for further development of OMV as a vaccination strategy in malaria and other diseases.

  2. [(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.

  3. 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.

  4. 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...

  5. Development of myiasis vaccine: In vitro detection of immunoprotective responses of peritrophic membrane protein, first instar larva Ll supernatant and pellet antigen of fly Chrysomyia bezziana in sheep

    Directory of Open Access Journals (Sweden)

    Sukarsih

    1999-10-01

    Full Text Available Myiasis control by means of individual treatment of animals which are mainly rised extensively is time consumed and expensive. The alternative way to control this disease by vaccination is considered effective and economically accepted. However the expected vaccine is now still being developed under a collaborative project between CSIRO, Inter-University Centre on Biotechnology-ITB and Research Institute for Veterinary Science and funded by ACIAR. There are several antigens have been identified as vaccine candidates and an in vitro bioassay technique has been developed for assessing the immunoresponses of vaccine in sheep. Three antigens were used for vaccines in this study, these included protein peritrophic membrane (PM, soluble extract (SE and pellet extract (PE of 1st instar larvae of Chrysomya bezziana. Twenty four experimental sheep were divided into 4 groups of 6 animals, 3 groups of animals were injected with PM, SE and PE vaccines with the dose rate of 0.5 g PM/head, 0.8 g PE/head and 4.2 ml LE/head respectively, and the other one group was injected with 4 ml PBS/head as a control group. Vaccination with the same dose was repeated 4 weeks after the 1st vaccination as a booster, and 2 weeks after the booster the sheep were challenged with live larvae, 3 days after challenge animals were killed. Sera were collected at the day of vaccination, 4 weeks after vaccination, 2 weeks after booster, and 3 days after challenge. An in vitro bioassay technique was conducted by culturing 1st instar larvae on five media containing sera collected from each experimental animal. The effects of sera on cultivated larvae were assessed by means of larval weight and larval mortality rate. The results indicated that the growth rate and survival of cultivated larvae in media containing anti-PM sera were significantly lower (P<0.01 compared to the larvae cultivated on media with sera on the day of vaccination. The larval weight depression by anti- PM sera

  6. 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. Copyright © 2015 Elsevier Ltd. All rights reserved.

  7. Integrated 68Gallium Labelled Prostate-Specific Membrane Antigen-11 Positron Emission Tomography/Magnetic Resonance Imaging Enhances Discriminatory Power of Multi-Parametric Prostate Magnetic Resonance Imaging.

    Science.gov (United States)

    Al-Bayati, Mohammad; Grueneisen, Johannes; Lütje, Susanne; Sawicki, Lino M; Suntharalingam, Saravanabavaan; Tschirdewahn, Stephan; Forsting, Michael; Rübben, Herbert; Herrmann, Ken; Umutlu, Lale; Wetter, Axel

    2018-01-01

    To evaluate diagnostic accuracy of integrated 68Gallium labelled prostate-specific membrane antigen (68Ga-PSMA)-11 positron emission tomography (PET)/MRI in patients with primary prostate cancer (PCa) as compared to multi-parametric MRI. A total of 22 patients with recently diagnosed primary PCa underwent clinically indicated 68Ga-PSMA-11 PET/CT for initial staging followed by integrated 68Ga-PSMA-11 PET/MRI. Images of multi-parametric magnetic resonance imaging (mpMRI), PET and PET/MRI were evaluated separately by applying Prostate Imaging Reporting and Data System (PIRADSv2) for mpMRI and a 5-point Likert scale for PET and PET/MRI. Results were compared with pathology reports of biopsy or resection. Statistical analyses including receiver operating characteristics analysis were performed to compare the diagnostic performance of mpMRI, PET and PET/MRI. PET and integrated PET/MRI demonstrated a higher diagnostic accuracy than mpMRI (area under the curve: mpMRI: 0.679, PET and PET/MRI: 0.951). The proportion of equivocal results (PIRADS 3 and Likert 3) was considerably higher in mpMRI than in PET and PET/MRI. In a notable proportion of equivocal PIRADS results, PET led to a correct shift towards higher suspicion of malignancy and enabled correct lesion classification. Integrated 68Ga-PSMA-11 PET/MRI demonstrates higher diagnostic accuracy than mpMRI and is particularly valuable in tumours with equivocal results from PIRADS classification. © 2018 S. Karger AG, Basel.

  8. Highly Efficient Capture and Enumeration of Low Abundance Prostate Cancer Cells Using Prostate-Specific Membrane Antigen Aptamers Immobilized to a Polymeric Microfluidic Device

    Science.gov (United States)

    Dharmasiri, Udara; Balamurugan, Subramanian; Adams, André A.; Okagbare, Paul I.; Obubuafo, Annie; Soper, Steven A.

    2010-01-01

    Prostate tumor cells over-express a prostate specific membrane antigen (PSMA) that can be used as a marker to select these cells from highly heterogeneous clinical samples, even when found in low abundance. Antibodies and aptamers have been developed that specifically bind to PSMA. In this study, anti-PSMA aptamers were immobilized onto the surface of a capture bed poised within a poly(methyl methacrylate), PMMA, microchip, which was fabricated into a high throughput micro-sampling unit (HTMSU) used for the selective isolation of rare circulating prostate tumor cells resident in a peripheral blood matrix. The HTMSU capture bed consisted of 51 ultra-high aspect ratio parallel curvilinear channels with a width similar to the prostate cancer cell dimensions. The surface density of the PSMA-specific aptamers on a UV-modified PMMA microfluidic capture bed surface was determined to be 8.4 × 1012 molecules/cm2. Using a linear velocity for optimal cell capture in the aptamer-tethered HTMSU (2.5 mm/s), a recovery of 90% of LNCaP cells (prostate cancer cell line; used as a model in this example) was found. Due to the low abundance of these cells, the input volume required was 1 mL and this could be processed in approximately 29 min using an optimized linear flow rate of 2.5 mm/s. Captured cells were subsequently released intact from the affinity surface using 0.25% (w/v) trypsin followed by counting individual cells using a contact conductivity sensor integrated into the HTMSU that provided high detection and sampling efficiency (~100%) and did not require staining of the cells for enumeration. PMID:19722212

  9. Humanised IgG1 antibody variants targeting membrane-bound carcinoembryonic antigen by antibody-dependent cellular cytotoxicity and phagocytosis.

    Science.gov (United States)

    Ashraf, S Q; Umana, P; Mössner, E; Ntouroupi, T; Brünker, P; Schmidt, C; Wilding, J L; Mortensen, N J; Bodmer, W F

    2009-11-17

    The effect of glycoengineering a membrane specific anti-carcinoembryonic antigen (CEA) (this paper uses the original term CEA for the formally designated CEACAM5) antibody (PR1A3) on its ability to enhance killing of colorectal cancer (CRC) cell lines by human immune effector cells was assessed. In vivo efficacy of the antibody was also tested. The antibody was modified using EBNA cells cotransfected with beta-1,4-N-acetylglucosaminyltransferase III and the humanised hPR1A3 antibody genes. The resulting alteration of the Fc segment glycosylation pattern enhances the antibody's binding affinity to the FcgammaRIIIa receptor on human immune effector cells but does not alter the antibody's binding capacity. Antibody-dependent cellular cytotoxicity (ADCC) is inhibited in the presence of anti-FcgammaRIII blocking antibodies. This glycovariant of hPR1A3 enhances ADCC 10-fold relative to the parent unmodified antibody using either unfractionated peripheral blood mononuclear or natural killer (NK) cells and CEA-positive CRC cells as targets. NK cells are far more potent in eliciting ADCC than either freshly isolated monocytes or granulocytes. Flow cytometry and automated fluorescent microscopy have been used to show that both versions of hPR1A3 can induce antibody-dependent cellular phagocytosis (ADCP) by monocyte-derived macrophages. However, the glycovariant antibody did not mediate enhanced ADCP. This may be explained by the relatively low expression of FcgammaRIIIa on cultured macrophages. In vivo studies show the efficacy of glycoengineered humanised IgG1 PR1A3 in significantly improving survival in a CRC metastatic murine model. The greatly enhanced in vitro ADCC activity of the glycoengineered version of hPR1A3 is likely to be clinically beneficial.

  10. Different immunogenicity but similar antitumor efficacy of two DNA vaccines coding for an antigen secreted in different membrane vesicle-associated forms

    OpenAIRE

    Bellier, Bertrand; Sedlik, Christine; Vigneron, James; Torrieri-Dramard, Lea; Pitoiset, Fabien; Denizeau, Jordan; Chesneau, Caroline; de la Rochere, Philippe; Lantz, Olivier; Thery, Clotilde

    2014-01-01

    The induction of an active immune response to control or eliminate tumours is still an unfulfilled challenge. We focused on plasmid DNA vaccines using an innovative approach whereby the antigen is expressed in association with extracellular vesicles (EVs) to facilitate antigen cross-presentation and improve induced immunity. Our two groups had independently shown previously that DNA vaccines encoding EV-associated antigens are more efficient at inducing cytotoxic T-cell responses than vaccine...

  11. Tolerance of brightness and contrast adjustments on chronic apical abscess and apical granuloma interpretation

    Science.gov (United States)

    Purnamasari, L.; Iskandar, H. H. B.; Makes, B. N.

    2017-08-01

    In digitized radiography techniques, adjusting the image enhancement can improve the subjective image quality by optimizing the brightness and contrast for diagnostic needs. To determine the value range of image enhancement (brightness and contrast) on chronic apical abscess and apical granuloma interpretation. 30 periapical radiographs that diagnosed chronic apical abscess and 30 that diagnosed apical granuloma were adjusted by changing brightness and contrast values. The value range of brightness and contrast adjustment that can be tolerated in radiographic interpretations of chronic apical abscess and apical granuloma spans from -10 to +10. Brightness and contrast adjustments on digital radiographs do not affect the radiographic interpretation of chronic apical abscess and apical granuloma if conducted within the value range.

  12. Correlation between prostate brachytherapy-related urethral stricture and peri-apical urethral dosimetry: A matched case–control study

    International Nuclear Information System (INIS)

    Earley, James J.; Abdelbaky, Ather M.; Cunningham, Melanie J.; Chadwick, Eliot; Langley, Stephen E.M.; Laing, Robert W.

    2012-01-01

    Background and purpose: Radiation dose to the bulbomembranous urethra has been shown to correlate with urethral stricture formation. This retrospective case–control study was designed to explore the relationship between dose to the apical/peri-apical regions of the urethra and development of brachytherapy (BXT)-related urethral stricture. Materials and methods: Cases were patients who developed urethral stricture after treatment with BXT as monotherapy and who had urethral dosimetry post-implant. Each case was matched with a control that had not developed urethral stricture. Dosimetry was compared between cases and controls. Results: Twenty-three cases were pair matched with 23 controls. There were no significant differences between the two groups in terms of age, presenting Prostate Specific Antigen (PSA), International Prostate Symptom Score (IPSS) or Gleason score. The dose delivered to the peri-apical and apical urethra was significantly higher for cases when compared with controls (peri-apical urethra: mean V 150 1.1 Vs 0.8 cc [p = 0.02]; apical urethra: mean dose 200 Vs 174 Gy [p = 0.01]). The distance from the prostate apex to isodose lines was also found to be significant in predicting stricture formation. Conclusion: There was evidence to suggest that the development of BXT-related stricture was associated with radiation dose at the apical and peri-apical urethra. Attention to the dose delivered to those areas may minimise the risk of developing such morbidity.

  13. Neuroelectric Tuning of Cortical Oscillations by Apical Dendrites in Loop Circuits

    Directory of Open Access Journals (Sweden)

    David LaBerge

    2017-06-01

    Full Text Available Bundles of relatively long apical dendrites dominate the neurons that make up the thickness of the cerebral cortex. It is proposed that a major function of the apical dendrite is to produce sustained oscillations at a specific frequency that can serve as a common timing unit for the processing of information in circuits connected to that apical dendrite. Many layer 5 and 6 pyramidal neurons are connected to thalamic neurons in loop circuits. A model of the apical dendrites of these pyramidal neurons has been used to simulate the electric activity of the apical dendrite. The results of that simulation demonstrated that subthreshold electric pulses in these apical dendrites can be tuned to specific frequencies and also can be fine-tuned to narrow bandwidths of less than one Hertz (1 Hz. Synchronous pulse outputs from the circuit loops containing apical dendrites can tune subthreshold membrane oscillations of neurons they contact. When the pulse outputs are finely tuned, they function as a local “clock,” which enables the contacted neurons to synchronously communicate with each other. Thus, a shared tuning frequency can select neurons for membership in a circuit. Unlike layer 6 apical dendrites, layer 5 apical dendrites can produce burst firing in many of their neurons, which increases the amplitude of signals in the neurons they contact. This difference in amplitude of signals serves as basis of selecting a sub-circuit for specialized processing (e.g., sustained attention within the typically larger layer 6-based circuit. After examining the sustaining of oscillations in loop circuits and the processing of spikes in network circuits, we propose that cortical functioning can be globally viewed as two systems: a loop system and a network system. The loop system oscillations influence the network system’s timing and amplitude of pulse signals, both of which can select circuits that are momentarily dominant in cortical activity.

  14. Macrophage polarization differs between apical granulomas, radicular cysts, and dentigerous cysts.

    Science.gov (United States)

    Weber, Manuel; Schlittenbauer, Tilo; Moebius, Patrick; Büttner-Herold, Maike; Ries, Jutta; Preidl, Raimund; Geppert, Carol-Immanuel; Neukam, Friedrich W; Wehrhan, Falk

    2018-01-01

    Apical periodontitis can appear clinically as apical granulomas or radicular cysts. There is evidence that immunologic factors are involved in the pathogenesis of both pathologies. In contrast to radicular cysts, the dentigerous cysts have a developmental origin. Macrophage polarization (M1 vs M2) is a main regulator of tissue homeostasis and differentiation. There are no studies comparing macrophage polarization in apical granulomas, radicular cysts, and dentigerous cysts. Forty-one apical granulomas, 23 radicular cysts, and 23 dentigerous cysts were analyzed in this study. A tissue microarray (TMA) of the 87 consecutive specimens was created, and CD68-, CD11c-, CD163-, and MRC1-positive macrophages were detected by immunohistochemical methods. TMAs were digitized, and the expression of macrophage markers was quantitatively assessed. Radicular cysts are characterized by M1 polarization of macrophages while apical granulomas show a significantly higher degree of M2 polarization. Dentigerous cysts have a significantly lower M1 polarization than both analyzed periapical lesions (apical granulomas and radicular cysts) and accordingly, a significantly higher M2 polarization than radicular cysts. Macrophage cell density in dentigerous cysts is significantly lower than in the periapical lesions. The development of apical periodontitis towards apical granulomas or radicular cysts might be directed by macrophage polarization. Radicular cyst formation is associated with an increased M1 polarization of infiltrating macrophages. In contrast to radicular cysts, dentigerous cysts are characterized by a low macrophage infiltration and a high degree of M2 polarization, possibly reflecting their developmental rather than inflammatory origin. As M1 polarization of macrophages is triggered by bacterial antigens, these results underline the need for sufficient bacterial clearance during endodontic treatment to prevent a possible M1 macrophage-derived stimulus for radicular cyst

  15. Lu-177-PSMA-617 Prostate-Specific Membrane Antigen Inhibitor Therapy in Patients with Castration-Resistant Prostate Cancer: Stability, Bio-distribution and Dosimetry

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    Levent Kabasakal

    2017-06-01

    Full Text Available Objective: The aim of the study was to estimate the radiation-absorbed doses and to study the in vivo and in vitro stability as well as pharmacokinetic characteristics of lutetium-177 (Lu-177 prostate-specific membrane antigen (PSMA-617. Methods: For this purpose, 7 patients who underwent Lu-177-PSMA therapy were included into the study. The injected Lu-177-PSMA-617 activity ranged from 3.6 to 7.4 GBq with a mean of 5.2±1.8 GBq. The stability of radiotracer in saline was calculated up to 48 h. The stability was also calculated in blood and urine samples. Post-therapeutic dosimetry was performed based on whole body and single photon emission computed tomography/computed tomography (SPECT/CT scans on dual-headed SPECT/CT system. Results: The radiochemical yield of Lu-177-PSMA-617 was >99%. It remained stable in saline up to 48 h. Analyses of the blood and urine samples showed a single radioactivity peak even at 24 hours after injection. Half-life of the distribution and elimination phases were calculated to be 0.16±0.09 and 10.8±2.5 hours, respectively. The mean excretion rate was 56.5±8.8% ranging from 41.5% to 65.4% at 24 h. Highest radiation estimated doses were calculated for parotid glands and kidneys (1.90±1.19 and 0.82±0.25 Gy/GBq respectively. Radiation dose given to the bone marrow was significantly lower than those of kidney and parotid glands (p<0.05 (0.030±0.008 Gy/GBq. Conclusion: Lu-177-PSMA-617 is a highly stable compound both in vitro and in vivo. Lu-177-PSMA-617 therapy seems to be a safe method for the treatment of castration-resistant prostate cancer patients. The fractionation regime that enables the longest duration of tumor control and/or survival will have to be developed in further studies.

  16. 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

  17. Intranasal Immunization with Influenza Virus-Like Particles Containing Membrane-Anchored Cholera Toxin B or Ricin Toxin B Enhances Adaptive Immune Responses and Protection against an Antigenically Distinct Virus.

    Science.gov (United States)

    Ji, Xianliang; Ren, Zhiguang; Xu, Na; Meng, Lingnan; Yu, Zhijun; Feng, Na; Sang, Xiaoyu; Li, Shengnan; Li, Yuanguo; Wang, Tiecheng; Zhao, Yongkun; Wang, Hualei; Zheng, Xuexing; Jin, Hongli; Li, Nan; Yang, Songtao; Cao, Jinshan; Liu, Wensen; Gao, Yuwei; Xia, Xianzhu

    2016-04-21

    Vaccination is the most effective means to prevent influenza virus infection, although current approaches are associated with suboptimal efficacy. Here, we generated virus-like particles (VLPs) composed of the hemagglutinin (HA), neuraminidase (NA) and matrix protein (M1) of A/Changchun/01/2009 (H1N1) with or without either membrane-anchored cholera toxin B (CTB) or ricin toxin B (RTB) as molecular adjuvants. The intranasal immunization of mice with VLPs containing membrane-anchored CTB or RTB elicited stronger humoral and cellular immune responses when compared to mice immunized with VLPs alone. Administration of VLPs containing CTB or RTB significantly enhanced virus-specific systemic and mucosal antibody responses, hemagglutination inhibiting antibody titers, virus neutralizing antibody titers, and the frequency of virus-specific IFN-γ and IL-4 secreting splenocytes. VLPs with and without CTB or RTB conferred complete protection against lethal challenge with a mouse-adapted homologous virus. When challenged with an antigenically distinct H1N1 virus, all mice immunized with VLPs containing CTB or RTB survived whereas mice immunized with VLPs alone showed only partial protection (80% survival). Our results suggest that membrane-anchored CTB and RTB possess strong adjuvant properties when incorporated into an intranasally-delivered influenza VLP vaccine. Chimeric influenza VLPs containing CTB or RTB may represent promising vaccine candidates for improved immunological protection against homologous and antigenically distinct influenza viruses.

  18. Intranasal Immunization with Influenza Virus-Like Particles Containing Membrane-Anchored Cholera Toxin B or Ricin Toxin B Enhances Adaptive Immune Responses and Protection against an Antigenically Distinct Virus

    Directory of Open Access Journals (Sweden)

    Xianliang Ji

    2016-04-01

    Full Text Available Vaccination is the most effective means to prevent influenza virus infection, although current approaches are associated with suboptimal efficacy. Here, we generated virus-like particles (VLPs composed of the hemagglutinin (HA, neuraminidase (NA and matrix protein (M1 of A/Changchun/01/2009 (H1N1 with or without either membrane-anchored cholera toxin B (CTB or ricin toxin B (RTB as molecular adjuvants. The intranasal immunization of mice with VLPs containing membrane-anchored CTB or RTB elicited stronger humoral and cellular immune responses when compared to mice immunized with VLPs alone. Administration of VLPs containing CTB or RTB significantly enhanced virus-specific systemic and mucosal antibody responses, hemagglutination inhibiting antibody titers, virus neutralizing antibody titers, and the frequency of virus-specific IFN-γ and IL-4 secreting splenocytes. VLPs with and without CTB or RTB conferred complete protection against lethal challenge with a mouse-adapted homologous virus. When challenged with an antigenically distinct H1N1 virus, all mice immunized with VLPs containing CTB or RTB survived whereas mice immunized with VLPs alone showed only partial protection (80% survival. Our results suggest that membrane-anchored CTB and RTB possess strong adjuvant properties when incorporated into an intranasally-delivered influenza VLP vaccine. Chimeric influenza VLPs containing CTB or RTB may represent promising vaccine candidates for improved immunological protection against homologous and antigenically distinct influenza viruses.

  19. 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.

  20. A Novel ¹¹¹In-Labeled Anti-Prostate-Specific Membrane Antigen Nanobody for Targeted SPECT/CT Imaging of Prostate Cancer.

    Science.gov (United States)

    Chatalic, Kristell L S; Veldhoven-Zweistra, Joke; Bolkestein, Michiel; Hoeben, Sander; Koning, Gerben A; Boerman, Otto C; de Jong, Marion; van Weerden, Wytske M

    2015-07-01

    Prostate-specific membrane antigen (PSMA) is overexpressed in prostate cancer (PCa) and a promising target for molecular imaging and therapy. Nanobodies (single-domain antibodies, VHH) are the smallest antibody-based fragments possessing ideal molecular imaging properties, such as high target specificity and rapid background clearance. We developed a novel anti-PSMA Nanobody (JVZ-007) for targeted imaging and therapy of PCa. Here, we report on the application of the (111)In-radiolabeled Nanobody for SPECT/CT imaging of PCa. A Nanobody library was generated by immunization of a llama with 4 human PCa cell lines. Anti-PSMA Nanobodies were captured by biopanning on PSMA-overexpressing cells. JVZ-007 was selected for evaluation as an imaging probe. JVZ-007 was initially produced with a c-myc-hexahistidine (his) tag allowing purification and detection. The c-myc-his tag was subsequently replaced by a single cysteine at the C terminus, allowing site-specific conjugation of chelates for radiolabeling. JVZ-007-c-myc-his was conjugated to 2-(4-isothiocyanatobenzyl)-diethylenetriaminepentaacetic acid (p-SCN-DTPA) via the lysines, whereas JVZ-007-cys was conjugated to maleimide-DTPA via the C-terminal cysteine. PSMA targeting was analyzed in vitro by cell-binding experiments using flow cytometry, autoradiography, and internalization assays with various PCa cell lines and patient-derived xenografts (PDXs). The targeting properties of radiolabeled Nanobodies were evaluated in vivo in biodistribution and SPECT/CT imaging experiments, using nude mice bearing PSMA-positive PC-310 and PSMA-negative PC-3 tumors. JVZ-007 was successfully conjugated to DTPA for radiolabeling with (111)In at room temperature. (111)In-JVZ007-c-myc-his and (111)In-JVZ007-cys internalized in LNCaP cells and bound to PSMA-expressing PDXs and, importantly, not to PSMA-negative PDXs and human kidneys. Good tumor targeting and fast blood clearance were observed for (111)In-JVZ-007-c-myc-his and (111)In

  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

    -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...... 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...... commonly recognized PfEMP1 proteins (HB3VAR06 and IT4VAR60) implicated in rosetting and not pregnancy restricted. This enabled, for the first time, a direct comparison in the same individuals of immune responses specific for a clinically important parasite antigen expressed only during well-defined periods...

  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 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.

  4. Stratification of Antigen-presenting Cells within the Normal Cornea

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    Jared E. Knickelbein

    2009-11-01

    Full Text Available The composition and location of professional antigen presenting cells (APC varies in different mucosal surfaces. The cornea, long considered an immune-privileged tissue devoid of APCs, is now known to host a heterogeneous network of bone marrow-derived cells. Here, we utilized transgenic mice that express enhanced green fluorescent protein (EGFP from the CD11c promoter (pCD11c in conjunction with immunohistochemical staining to demonstrate an interesting stratification of APCs within non-inflamed murine corneas. pCD11c+ dendritic cells (DCs reside in the basal epithelium, seemingly embedded in the basement membrane. Most DCs express MHC class II on at least some dendrites, which extend up to 50 µm in length and traverse up 20 µm tangentially towards the apical surface of the epithelium. The DC density diminishes from peripheral to central cornea. Beneath the DCs and adjacent to the stromal side of the basement membrane reside pCD11c-CD11b+ putative macrophages that express low levels of MHC class II. Finally, MHC class IIpCD11c-CD11b+ cells form a network throughout the remainder of the stroma. This highly reproducible stratification of bone marrow-derived cells is suggestive of a progression from an APC function at the exposed corneal surface to an innate immune barrier function deeper in the stroma.

  5. 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.

  6. Apical dominance and apical control in multiple flushing of temperate woody species.

    Science.gov (United States)

    M. Cline; C. Harrington

    2007-01-01

    In young plants of many woody species, the first flush of growth in the spring may be followed by one or more flushes of the terminal shoot if growing conditions are favorable. The occurrence of these additional flushes may significantly affect crown form and structure. Apical dominance (AD) and apical control (AC) are thought to be important control mechanisms in this...

  7. 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

  8. Microbiology and Treatment of Acute Apical Abscesses

    Science.gov (United States)

    Rôças, Isabela N.

    2013-01-01

    SUMMARY Acute apical abscess is the most common form of dental abscess and is caused by infection of the root canal of the tooth. It is usually localized intraorally, but in some cases the apical abscess may spread and result in severe complications or even mortality. The reasons why dental root canal infections can become symptomatic and evolve to severe spreading and sometimes life-threatening abscesses remain elusive. Studies using culture and advanced molecular microbiology methods for microbial identification in apical abscesses have demonstrated a multispecies community conspicuously dominated by anaerobic bacteria. Species/phylotypes commonly found in these infections belong to the genera Fusobacterium, Parvimonas, Prevotella, Porphyromonas, Dialister, Streptococcus, and Treponema. Advances in DNA sequencing technologies and computational biology have substantially enhanced the knowledge of the microbiota associated with acute apical abscesses and shed some light on the etiopathogeny of this disease. Species richness and abundance and the resulting network of interactions among community members may affect the collective pathogenicity and contribute to the development of acute infections. Disease modifiers, including transient or permanent host-related factors, may also influence the development and severity of acute abscesses. This review focuses on the current evidence about the etiology and treatment of acute apical abscesses and how the process is influenced by host-related factors and proposes future directions in research, diagnosis, and therapeutic approaches to deal with this disease. PMID:23554416

  9. Incidental apical disease at CT scanning

    International Nuclear Information System (INIS)

    McLoud, T.C.; Satoh, K.; Shepard, J.O.; Moore, E.H.; Kosiuk, J.P.

    1990-01-01

    Apical caps are commonly noted on standard radiographs. This paper determines how often abnormalities in the extreme apex of the lung could be identified on CT scans obtained for other reasons. A total of 158 consecutive CT scans were reviewed prospectively. Excluded were patients with obvious upper lobe pleural or parenchymal disease. Apical abnormalities were identified in 74 (46.8%) of the 158 cases. The prevalence increased with age (19% in the 8-39-year age group and 82% in patients older than 80 years). Opacities were unilateral in 44.5% and bilateral in 55.5%. The most common abnormality was linear opacities (95%)

  10. Relationship between the Apical Preparation Diameter and the Apical Seal: An In Vitro Study

    Directory of Open Access Journals (Sweden)

    Kaoutar Laslami

    2018-01-01

    Full Text Available Objectives. The aim of the study is to define the relationship between the apical preparation diameter and the apical sealing ability to highlight the importance of the preservation of the diameter and the original position of the apical foramen. Materials and Methods. 50 extracted maxillary incisors were randomly allocated into three groups of 15 teeth each (n = 15 according to the apical preparation size: Group 1: finishing file F1 corresponding to size 20 reached the working length (ProTaper Universal system Dentsply®; Group 2: prepared up to size 30 corresponding to finishing file F30; Group 3: prepared up to size 50 corresponding to finishing file F5. Five teeth were assigned to positive and negative control groups. After the filling of the root canals, the teeth were isolated and immersed in a dye solution, then cut longitudinally, photographed, and the dye penetration were calculated using a computer software. Results. Comparison of the three different apical preparation sizes showed no statistically significant differences regarding the apical microleakage. Conclusion. The most important value of the dye penetration was observed in the group with the largest apical diameter.

  11. 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...

  12. 19-DEJ-1, a hemidesmosome-anchoring filament complex-associated monoclonal antibody. Definition of a new skin basement membrane antigenic defect in junctional and dystrophic epidermolysis bullosa

    DEFF Research Database (Denmark)

    Fine, J D; Horiguchi, Y; Couchman, J R

    1989-01-01

    50% of patients with recessive dystrophic EB further suggests, for the first time, that an inherited defect in basement membrane is shared in patients with junctional and some recessive dystrophic EB. The difference in site of skin cleavage in the latter two disorders gives additional support...

  13. 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

    Human Leukocyte Antigen (HLA)-G is an immunosuppressive molecule acting on both the innate and adaptive immune system. A 14 bp insertion/deletion polymorphism (rs66554220) in the 3'-untranslated region (3'UTR) of the HLA-G gene has been associated with a number of diseases, pregnancy complications...... insertion were the most efficient in inhibiting NK cytotoxicity but showed a lower soluble HLA-G1 per mHLA-G1 ratio than the HLA-G1 K562 cells lacking the 14 bp insertion. Our data suggest 3'UTR polymorphism may play an important role in HLA-G regulation with implications on a range of diseases....

  14. Apical Hypertrophic Cardiomyopathy Mimicking Acute Coronary Syndrome

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    Lütfü Bekar

    2013-03-01

    Full Text Available Apical hypertrophic cardiomyopathy is a rare form of hypertrophic cardiomyopathy and it can be mistaken for coronary artery disease due to the symptoms and electrocardiography findings. In this report, we aimed to present a patient referred to our clinic with complaints of chest pain and electrocardiography findings who had been misdiagnosed as non-ST elevation acute coronary syndrome.

  15. The resection angle in apical surgery

    DEFF Research Database (Denmark)

    von Arx, Thomas; Janner, Simone F M; Jensen, Simon S

    2016-01-01

    OBJECTIVES: The primary objective of the present radiographic study was to analyse the resection angle in apical surgery and its correlation with treatment outcome, type of treated tooth, surgical depth and level of root-end filling. MATERIALS AND METHODS: In the context of a prospective clinical...

  16. Pathogenesis of Apical Periodontitis: a Literature Review

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    Indre Graunaite

    2011-09-01

    Full Text Available Objectives: This review article discusses the host response in apical periodontitis with the main focus on cytokines, produced under this pathological condition and contributing to the degradation of periradicular tissues. The pace of research in this field has greatly accelerated in the last decade. Here we provide an analysis of studies published in this area during this period.Material and methods: Literature was selected through a search of PubMed electronic database. The keywords used for search were pathogenesis of apical periodontitis cytokines, periapical granuloma cytokines, inflammatory infiltrate apical periodontitis. The search was restricted to English language articles, published from 1999 to December 2010. Additionally, a manual search in the cytokine production, cytokine functions and periapical tissue destruction in the journals and books was performed.Results: In total, 97 literature sources were obtained and reviewed. The topics covered in this article include cellular composition of an inflammatory infiltrate in the periapical lesions, mechanisms of the formation of the innate and specific immune response. Studies which investigated cytokine secretion and functions were identified and cellular and molecular interactions in the course of apical periodontitis described.Conclusions: The abundance and interactions of various inflammatory and anti-inflammatory molecules can influence and alter the state and progression of the disease. Therefore, periapical inflammatory response offers a model, suited for the study of many facets of pathogenesis, biocompatibility of different materials to periapical tissues and development of novel treatment methods, based on the regulation of cytokines expression.

  17. Pathogenesis of apical periodontitis: a literature review.

    Science.gov (United States)

    Graunaite, Indre; Lodiene, Greta; Maciulskiene, Vita

    2012-01-01

    This review article discusses the host response in apical periodontitis with the main focus on cytokines, produced under this pathological condition and contributing to the degradation of periradicular tissues. The pace of research in this field has greatly accelerated in the last decade. Here we provide an analysis of studies published in this area during this period. Literature was selected through a search of PubMed electronic database. The keywords used for search were pathogenesis of apical periodontitis cytokines, periapical granuloma cytokines, inflammatory infiltrate apical periodontitis. The search was restricted to English language articles, published from 1999 to December 2010. Additionally, a manual search in the cytokine production, cytokine functions and periapical tissue destruction in the journals and books was performed. In total, 97 literature sources were obtained and reviewed. The topics covered in this article include cellular composition of an inflammatory infiltrate in the periapical lesions, mechanisms of the formation of the innate and specific immune response. Studies which investigated cytokine secretion and functions were identified and cellular and molecular interactions in the course of apical periodontitis described. The abundance and interactions of various inflammatory and anti-inflammatory molecules can influence and alter the state and progression of the disease. Therefore, periapical inflammatory response offers a model, suited for the study of many facets of pathogenesis, biocompatibility of different materials to periapical tissues and development of novel treatment methods, based on the regulation of cytokines expression.

  18. Using the AD12-ICT rapid-format test to detect Wuchereria bancrofti circulating antigens in comparison to Og4C3-ELISA and nucleopore membrane filtration and microscopy techniques.

    Science.gov (United States)

    El-Moamly, Amal Abdul-Rasheed; El-Sweify, Mohamed Aly; Hafez, Mohamad Abdul

    2012-09-01

    Lymphatic filariasis (LF) continues to be a major source of permanent disability and an impediment to socio-economic development in 73 countries where more than 1 billion people are at risk and over 120 millions are infected. The global drive to eliminate LF necessitates an increasing demand for valid, reliable and rapid diagnostic tests. This study aimed to assess the performance of the AD12 rapid format immunochromatographic test (ICT) to detect Wuchereria bancrofti circulating antigens, against the combined gold standard: TropBio Og4C3-ELISA (enzyme-linked immunosorbent assay) which detects circulating filarial antigen (CFA) and the nucleopore membrane filtration and microscopic examination. This prospective case-control study involved 647 asymptomatic migrant workers from filariasis-endemic countries. Of these specimens, 32 were positive for microfilaremia using the membrane filtration and microscopy, 142 positive by ELISA (of which 32 had microfilaremia), and 128 positive by the ICT (of which 31 had microfilaremia). The performance of the ICT was calculated against 32 true-positive and 90 true-negative cases. For the detection of CFA, the ICT had a sensitivity of 97% (95% confidence interval [CI] 91-103), specificity 100% (95% CI 100-100), Positive Predictive Value (PPV) 100% (95% CI 100-100), Negative Predictive Value (NPV) 99% (95% CI 97-101); and the total accuracy of the test was 99% (95% CI 98-101). The agreement between ICT and ELISA in detecting W. bancrofti antigens was excellent (kappa = 0.934; p = 0.000). In conclusion, the AD12-ICT test for the detection of W. bancrofti-CFA was sensitive and specific and comparable to the performance of ELISA. The ICT would be a useful additional test to facilitate the proposed strategies for control and elimination of LF. Because it is rapid, simple to perform, and does not require the use of special equipment, the ICT may be most appropriate in screening programs and in monitoring the possible risk of introducing

  19. Apical versus Non-Apical Lead: Is ICD Lead Position Important for Successful Defibrillation?

    Science.gov (United States)

    Amit, Guy; Wang, Jia; Connolly, Stuart J; Glikson, Michael; Hohnloser, Stephan; Wright, David J; Brachmann, Johannes; Defaye, Pascal; Neuzner, Joerg; Mabo, Philippe; Vanerven, Liselot; Vinolas, Xavier; O'Hara, Gilles; Kautzner, Josef; Appl, Ursula; Gadler, Fredrik; Stein, Kenneth; Konstantino, Yuval; Healey, Jeff S

    2016-05-01

    We aim to compare the acute and long-term success of defibrillation between non-apical and apical ICD lead position. The position of the ventricular lead was recorded by the implanting physician for 2,475 of 2,500 subjects in the Shockless IMPLant Evaluation (SIMPLE) trial, and subjects were grouped accordingly as non-apical or apical. The success of intra-operative defibrillation testing and of subsequent clinical shocks were compared. Propensity scoring was used to adjust for the impact of differences in baseline variables between these groups. There were 541 leads that were implanted at a non-apical position (21.9%). Patients implanted with a non-apical lead had a higher rate of secondary prevention indication. Non-apical location resulted in a lower mean R-wave amplitude (14.0 vs. 15.2, P defibrillation was similar between propensity score matched groups (89%). Over a mean follow-up of 3 years, there were no significant differences in the yearly rates of appropriate shock (5.5% vs. 5.4%, P = 0.98), failed appropriate first shock (0.9% vs. 1.0%, P = 0.66), or the composite of failed shock or arrhythmic death (2.8% vs. 2.3% P = 0.35) according to lead location. We did not detect any reduction in the ICD efficacy at the time of implant or during follow-up in patients receiving a non-apical RV lead. © 2016 Wiley Periodicals, Inc.

  20. An internal deletion in the cytoplasmic tail reverses the apical localization of human NGF receptor in transfected MDCK cells.

    Science.gov (United States)

    Le Bivic, A; Sambuy, Y; Patzak, A; Patil, N; Chao, M; Rodriguez-Boulan, E

    1991-11-01

    A cDNA encoding the full-length 75-kD human nerve growth factor receptor was transfected into MDCK cells and its product was found to be expressed predominantly (80%) on the apical membrane, as a result of vectorial targeting from an intracellular site. Apical hNGFR bound NGF with low affinity and internalized it inefficiently (6% of surface bound NGF per hour). Several mutant hNGFRs were analyzed, after transfection in MDCK cells, for polarized surface expression, ligand binding, and endocytosis. Deletionof juxta-membrane attachment sites for a cluster of O-linked sugars did not alter apical localization. A mutant receptor lacking the entire cytoplasmic tail (except for the five proximal amino acids) was also expressed on the apical membrane, suggesting that information for apical sorting was contained in the ectoplasmic or transmembrane domains. However, a 58 amino acid deletion in the hNGFR tail that moved a cytoplasmic tyrosine (Tyr 308) closer to the membrane into a more charged environment resulted in a basolateral distribution of the mutant receptor and reversed vectorial (basolateral) targeting. The basolateral mutant receptor also internalized 125I-NGF rapidly (90% of surface bound NGF per hour), exhibited a larger intracellular fraction and displayed a considerably shortened half-life (approximately 3 h). We suggest that hNGFR with the internal cytoplasmic deletion expresses a basolateral targeting signal, related to endocytic signals, that is dominant over apical targeting information in the ecto/transmembrane domains. These results apparently contradict a current model that postulates that basolateral targeting is a default mechanism.

  1. A modern approach to surgical endodontics: ultrasonic apical preparation.

    Science.gov (United States)

    Kellert, M; Solomon, C; Chalfin, H

    1994-01-01

    Ultrasonic devices are capable of generating better apical preparations than rotary instruments with less sacrifice of root structure. Regardless of the technique used, good lighting and magnification are necessary to insure an adequate apical preparation.

  2. 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. © 2016 Wiley Periodicals, Inc.

  3. Efficacy Against Human Prostate Cancer by Prostate-specific Membrane Antigen-specific, Transforming Growth Factor-β Insensitive Genetically Targeted CD8+T-cells Derived from Patients with Metastatic Castrate-resistant Disease.

    Science.gov (United States)

    Zhang, Qiang; Helfand, Brian T; Carneiro, Benedito A; Qin, Weijun; Yang, Ximing J; Lee, Chung; Zhang, Weipeng; Giles, Francis J; Cristofanilli, Massimo; Kuzel, Timothy M

    2017-12-21

    Current immunotherapy has limited efficacy on metastatic castrate-resistant prostate cancer (mCRPC). We therefore sought to improve the antitumor ability of mCRPC patient-derived CD8 + T-cells by the endowment of specificity to prostate-specific membrane antigen (PSMA) and insensitivity to immunosuppressant molecule transforming growth factor-β (TGF-ß) under the control of herpes simplex virus-1 thymidine kinase. CD8 + T-cells were collected by leukapheresis and cultured in a Food and Drug Administration-approved Cell Processing Work Station. We developed a chimeric antigen receptor retroviral construct using an anti-PSMA chimeric immunoglobulin-T-cell receptor(ζ) gene (PZ1) and dominant negative TGF-ß type II receptor (TßRIIDN), that could induce CD8 + T-cells to be PSMA reactive and insensitive to TGF-ß. Cr 51 release assay was performed on PC-3 and PC-3-PSMA. The further antitumor functions of PSMA-specific, TGF-ß insensitive CD8 + T-cells was evaluated using an immunodeficient RAG-1 -/- mouse model. We found PSMA-specific, TGF-ß insensitive CD8 + T-cells from mCRPC were expanded with strong expression of PZ1 and thymidine kinase genes, and their growth was not suppressed by TGF-ß. The survival of these cells decreased sharply after treatment with ganciclovir. Treatment of PSMA-specific TGF-ß, insensitive CD8 + T-cells was associated with 61.58% specific lysis on PC-3-PSMA, and significantly suppressed PC3-PSMA tumor compared with the PC3 tumor. A large amount of tumor apoptosis and CD8 + T-cell infiltration were found only in the PC3-PSMA tumor. This study verified that PSMA-specific, TGF-ß insensitive CD8 + T-cells derived from mCRPC patients could be successfully expanded and used to overcome the immunosuppressive effects of the tumor microenvironment to control PSMA-expressing PC in vitro and in vivo. This may provide a promising approach for men with mCRPC who fail androgen deprivation therapy. We investigated the role of a novel chimeric antigen

  4. 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

    against placental malaria, VAR2CSA was studied extensively. By the use of SAXS on multiple truncated VAR2CSA constructs it was possible to combine the acquired information and present a structural model for the DBL and CIDR domain arrangement in VAR2CSA. This work was published in Journal of Biological...... symptoms and vaccine research is focused on trying to mimic or advance this immune acquisition. Immunity is primarily caused by acquisition of antibodies directed against a family of Plasmodium protein antigens called PfEMP1s located on the surface of infected erythrocytes. The PfEMP1 proteins are adhesive...... knowledge for these proteins is required. During the time course of this PhD thesis I have studied a number of PfEMP1 proteins using the SAXS technique. This biophysical technique has gained widespread attention during the past decade due to scientific and computational developments. SAXS is X...

  5. Regeneration of okra (Abelmoschus esculentus L.) via apical shoot ...

    African Journals Online (AJOL)

    USER

    2012-10-25

    Oct 25, 2012 ... The present study was undertaken to evaluate the most suitable concentration of growth regulators for regeneration of okra (Abelmoschus esculentus L. Monech) via apical shoot culture system. The study of apical shoot culture system was found effective for regeneration of apical shoots. The okra (A.

  6. Regeneration of okra ( Abelmoschus esculentus L.) via apical shoot ...

    African Journals Online (AJOL)

    The present study was undertaken to evaluate the most suitable concentration of growth regulators for regeneration of okra (Abelmoschus esculentus L. Monech) via apical shoot culture system. The study of apical shoot culture system was found effective for regeneration of apical shoots. The okra (A. esculentus L. Monech) ...

  7. 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 Caco-2 cells thus secrete uteroglobin in a non-sorted manner. It has, however, previously been shown that uteroglobin is secreted exclusively at the apical membrane in primary cell culture of endometrial epithelial cells [S.K. Mani et al. (1991) Endocrinology 128, 1563-1573]. This suggests that either...... 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....

  8. N-Glycosylation instead of cholesterol mediates oligomerization and apical sorting of GPI-APs in FRT cells.

    Science.gov (United States)

    Imjeti, Naga Salaija; Lebreton, Stéphanie; Paladino, Simona; de la Fuente, Erwin; Gonzalez, Alfonso; Zurzolo, Chiara

    2011-12-01

    Sorting of glycosylphosphatidyl-inositol--anchored proteins (GPI-APs) in polarized epithelial cells is not fully understood. Oligomerization in the Golgi complex has emerged as the crucial event driving apical segregation of GPI-APs in two different kind of epithelial cells, Madin-Darby canine kidney (MDCK) and Fisher rat thyroid (FRT) cells, but whether the mechanism is conserved is unknown. In MDCK cells cholesterol promotes GPI-AP oligomerization, as well as apical sorting of GPI-APs. Here we show that FRT cells lack this cholesterol-driven oligomerization as apical sorting mechanism. In these cells both apical and basolateral GPI-APs display restricted diffusion in the Golgi likely due to a cholesterol-enriched membrane environment. It is striking that N-glycosylation is the critical event for oligomerization and apical sorting of GPI-APs in FRT cells but not in MDCK cells. Our data indicate that at least two mechanisms exist to determine oligomerization in the Golgi leading to apical sorting of GPI-APs. One depends on cholesterol, and the other depends on N-glycosylation and is insensitive to cholesterol addition or depletion.

  9. 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.

  10. Evaluation of three instrumentation techniques at the precision of apical stop and apical sealing of obturation

    Directory of Open Access Journals (Sweden)

    Özgür Genç

    2011-08-01

    Full Text Available OBJECTIVE: The aim of this study was to investigate the ability of two NiTi rotary apical preparation techniques used with an electronic apex locator-integrated endodontic motor and a manual technique to create an apical stop at a predetermined level (0.5 mm short of the apical foramen in teeth with disrupted apical constriction, and to evaluate microleakage following obturation in such prepared teeth. MATERIAL AND METHODS: 85 intact human mandibular permanent incisors with single root canal were accessed and the apical constriction was disrupted using a #25 K-file. The teeth were embedded in alginate and instrumented to #40 using rotary Lightspeed or S-Apex techniques or stainless-steel K-files. Distance between the apical foramen and the created apical stop was measured to an accuracy of 0.01 mm. In another set of instrumented teeth, root canals were obturated using gutta-percha and sealer, and leakage was tested at 1 week and 3 months using a fluid filtration device. RESULTS: All techniques performed slightly short of the predetermined level. Closest preparation to the predetermined level was with the manual technique and the farthest was with S-Apex. A significant difference was found between the performances of these two techniques (p<0.05. Lightspeed ranked in between. Leakage was similar for all techniques at either period. However, all groups leaked significantly more at 3 months compared to 1 week (p<0.05. CONCLUSIONS: Despite statistically significant differences found among the techniques, deviations from the predetermined level were small and clinically acceptable for all techniques. Leakage following obturation was comparable in all groups.

  11. Pathogenesis of Apical Periodontitis: a Literature Review

    OpenAIRE

    Graunaite, Indre; Lodiene, Greta; Maciulskiene, Vita

    2012-01-01

    ABSTRACT Objectives This review article discusses the host response in apical periodontitis with the main focus on cytokines, produced under this pathological condition and contributing to the degradation of periradicular tissues. The pace of research in this field has greatly accelerated in the last decade. Here we provide an analysis of studies published in this area during this period. Material and methods Literature was selected through a search of PubMed electronic database. The keywords...

  12. Scanning electron microscopic study of apical and intracanal resorption.

    Science.gov (United States)

    Delzangles, B

    1989-07-01

    Apical radicular and intracanal surfaces of extracted teeth with apical lesions were examined by means of scanning electron microscopy. The distribution of apical and intracanal resorption areas varied with the presence of a granuloma or a cyst. Teeth bearing granulomas showed an apical resorption centered on the main foramina whereas the hard tissue underlying a cyst showed little or no resorption. Intracanal resorption was always marked in the apical third and more scattered in the middle and cervical third. The resorption disrupted the anatomical structures.

  13. A Plasmodium vivax Plasmid DNA- and Adenovirus-Vectored Malaria Vaccine Encoding Blood-Stage Antigens AMA1 and MSP142in a Prime/Boost Heterologous Immunization Regimen Partially Protects Aotus Monkeys against Blood-Stage Challenge.

    Science.gov (United States)

    Obaldia, Nicanor; Stockelman, Michael G; Otero, William; Cockrill, Jennifer A; Ganeshan, Harini; Abot, Esteban N; Zhang, Jianfeng; Limbach, Keith; Charoenvit, Yupin; Doolan, Denise L; Tang, De-Chu C; Richie, Thomas L

    2017-04-01

    Malaria is caused by parasites of the genus Plasmodium , which are transmitted to humans by the bites of Anopheles mosquitoes. After the elimination of Plasmodium falciparum , it is predicted that Plasmodium vivax will remain an important cause of morbidity and mortality outside Africa, stressing the importance of developing a vaccine against P. vivax malaria. In this study, we assessed the immunogenicity and protective efficacy of two P. vivax antigens, apical membrane antigen 1 (AMA1) and the 42-kDa C-terminal fragment of merozoite surface protein 1 (MSP1 42 ) in a plasmid recombinant DNA prime/adenoviral (Ad) vector boost regimen in Aotus monkeys. Groups of 4 to 5 monkeys were immunized with plasmid DNA alone, Ad alone, prime/boost regimens with each antigen, prime/boost regimens with both antigens, and empty vector controls and then subjected to blood-stage challenge. The heterologous immunization regimen with the antigen pair was more protective than either antigen alone or both antigens delivered with a single vaccine platform, on the basis of their ability to induce the longest prepatent period and the longest time to the peak level of parasitemia, the lowest peak and mean levels of parasitemia, the smallest area under the parasitemia curve, and the highest self-cure rate. Overall, prechallenge MSP1 42 antibody titers strongly correlated with a decreased parasite burden. Nevertheless, a significant proportion of immunized animals developed anemia. In conclusion, the P. vivax plasmid DNA/Ad serotype 5 vaccine encoding blood-stage parasite antigens AMA1 and MSP1 42 in a heterologous prime/boost immunization regimen provided significant protection against blood-stage challenge in Aotus monkeys, indicating the suitability of these antigens and this regimen for further development. Copyright © 2017 American Society for Microbiology.

  14. Ubiquitination of human leukocyte antigen (HLA)-DM by different membrane-associated RING-CH (MARCH) protein family E3 ligases targets different endocytic pathways.

    Science.gov (United States)

    Jahnke, Martin; Trowsdale, John; Kelly, Adrian P

    2012-03-02

    HLA-DM plays an essential role in the peptide loading of classical class II molecules and is present both at the cell surface and in late endosomal peptide-loading compartments. Trafficking of DM within antigen-presenting cells is complex and is, in part, controlled by a tyrosine-based targeting signal present in the cytoplasmic tail of DMβ. Here, we show that DM also undergoes post-translational modification through ubiquitination of a single lysine residue present in the cytoplasmic tail of the α chain, DMα. Ubiquitination of DM by MARCH1 and MARCH9 induced loss of DM molecules from the cell surface by a mechanism that cumulatively involved both direct attachment of ubiquitin chains to DMα and a functional tyrosine-based signal on DMβ. In contrast, MARCH8-induced loss of surface DM was entirely dependent upon the tyrosine signal on DMβ. In the absence of this tyrosine residue, levels of DM remained unchanged irrespective of whether DMα was ubiquitinated by MARCH8. The influence of MARCH8 was indirect and may have resulted from modification of components of the endocytic machinery by ubiquitination.

  15. Ubiquitination of Human Leukocyte Antigen (HLA)-DM by Different Membrane-associated RING-CH (MARCH) Protein Family E3 Ligases Targets Different Endocytic Pathways*

    Science.gov (United States)

    Jahnke, Martin; Trowsdale, John; Kelly, Adrian P.

    2012-01-01

    HLA-DM plays an essential role in the peptide loading of classical class II molecules and is present both at the cell surface and in late endosomal peptide-loading compartments. Trafficking of DM within antigen-presenting cells is complex and is, in part, controlled by a tyrosine-based targeting signal present in the cytoplasmic tail of DMβ. Here, we show that DM also undergoes post-translational modification through ubiquitination of a single lysine residue present in the cytoplasmic tail of the α chain, DMα. Ubiquitination of DM by MARCH1 and MARCH9 induced loss of DM molecules from the cell surface by a mechanism that cumulatively involved both direct attachment of ubiquitin chains to DMα and a functional tyrosine-based signal on DMβ. In contrast, MARCH8-induced loss of surface DM was entirely dependent upon the tyrosine signal on DMβ. In the absence of this tyrosine residue, levels of DM remained unchanged irrespective of whether DMα was ubiquitinated by MARCH8. The influence of MARCH8 was indirect and may have resulted from modification of components of the endocytic machinery by ubiquitination. PMID:22247549

  16. Quantitative RT-PCR analysis of PSA and prostate-specific membrane antigen mRNA to detect circulating tumor cells improves recurrence-free survival nomogram prediction after radical prostatectomy.

    Science.gov (United States)

    Yates, David R; Rouprêt, Morgan; Drouin, Sarah J; Comperat, Eva; Ricci, Sylvie; Lacave, Roger; Sèbe, Philippe; Cancel-Tassin, Geraldine; Bitker, Marc-Olivier; Cussenot, Olivier

    2012-09-01

    Circulating tumor cell (CTC) analysis is a potential new biomarker in prostate cancer. We hypothesize that quantitative detection of CTCs in patients pre- and post-radical prostatectomy (RP) using quantitative TaqMan® fluorogenic RT-PCR will improve the accuracy of the Kattan nomogram to predict the probability of recurrence-free survival (RFS) post-RP. Ninty-two patients who underwent RP between 2004 and 2009 had venous blood samples taken pre- (Day - 1) and post-operatively (Day + 7). We performed quantitative Taqman® RT-PCR to detect circulating prostate-specific antigen (PSA) and prostate-specific membrane antigen (PSMA) mRNA. We calculated both the logarithmic ratio of Day + 7/Day - 1 for PSA (PSAr) and PSMA (PSMAr) expression (log(Day+7/Day-1) ) and the Kattan nomogram predicted probability of disease recurrence for each patient. We then analyzed how the AUC-ROC analysis for the Kattan nomogram prediction alone (K) compared to the addition of the PSAr and PSMAr in predicting 5-year RFS. The mean age (years), PSA (ng/ml), and follow-up (mo) was 65.1, 9.13, and 72, respectively. The AUCs for K, PSAr + K, and PSMAr + K were 0.752 (95%CI 0.620-0.860), 0.830 (95%CI 0.740-0.911), and 0.837 (95%CI 0.613-0.923), respectively (P = 0.03). The Kattan 5-year PSA RFS was 75%. The actual 5-year PSA RFS survival rate was 77%. Data from modern quantitative RT-PCR to detect circulating prostate-derived PSA and PSM mRNA pre- and post-RP improves the accuracy of the Kattan nomogram to predict biochemical recurrence. Copyright © 2012 Wiley Periodicals, Inc.

  17. 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

  18. Rab11a is required for apical protein localisation in the intestine

    Directory of Open Access Journals (Sweden)

    Tomoaki Sobajima

    2014-12-01

    Full Text Available The small GTPase Rab11 plays an important role in the recycling of proteins to the plasma membrane as well as in polarised transport in epithelial cells and neurons. We generated conditional knockout mice deficient in Rab11a. Rab11a-deficient mice are embryonic lethal, and brain-specific Rab11a knockout mice show no overt abnormalities in brain architecture. In contrast, intestine-specific Rab11a knockout mice begin dying approximately 1 week after birth. Apical proteins in the intestines of knockout mice accumulate in the cytoplasm and mislocalise to the basolateral plasma membrane, whereas the localisation of basolateral proteins is unaffected. Shorter microvilli and microvillus inclusion bodies are also observed in the knockout mice. Elevation of a serum starvation marker was also observed, likely caused by the mislocalisation of apical proteins and reduced nutrient uptake. In addition, Rab8a is mislocalised in Rab11a knockout mice. Conversely, Rab11a is mislocalised in Rab8a knockout mice and in a microvillus atrophy patient, which has a mutation in the myosin Vb gene. Our data show an essential role for Rab11a in the localisation of apical proteins in the intestine and demonstrate functional relationships between Rab11a, Rab8a and myosin Vb in vivo.

  19. Systemic antibiotics for symptomatic apical periodontitis and acute apical abscess in adults.

    Science.gov (United States)

    Cope, Anwen; Francis, Nick; Wood, Fiona; Mann, Mala K; Chestnutt, Ivor G

    2014-06-26

    Dental pain can have a considerable detrimental effect on an individual's quality of life. Symptomatic apical periodontitis and acute apical abscess are common causes of dental pain and arise from an inflamed or necrotic dental pulp, or infection of the pulpless root canal system. Clinical guidelines recommend that the first-line treatment for teeth with symptomatic apical periodontitis or an acute apical abscess should be removal of the source of inflammation or infection by local, operative measures, and that systemic antibiotics are currently only recommended for situations where there is evidence of spreading infection (cellulitis, lymph node involvement, diffuse swelling) or systemic involvement (fever, malaise). Despite this, there is evidence that dentists continue to prescribe antibiotics for these conditions. There is concern that this could contribute to the development of antibiotic-resistant bacterial colonies both within the individual and within the community as a whole. To evaluate the effects of systemic antibiotics provided with or without surgical intervention (such as extraction, incision and drainage of a swelling or endodontic treatment), with or without analgesics, for symptomatic apical periodontitis or acute apical abscess in adults. We searched the following electronic databases: Cochrane Oral Health Group's Trials Register (to 1 October 2013); Cochrane Central Register of Controlled Trials (The Cochrane Library 2013, Issue 9); MEDLINE via OVID (1946 to 1 October 2013); EMBASE via OVID (1980 to 1 October 2013) and CINAHL via EBSCO (1980 to 1 October 2013). We searched the World Health Organization (WHO) International Trials Registry Platform and the US National Institutes of Health Trials Registry (ClinicalTrials.gov) on 1 October 2013 to identify ongoing trials. We searched for grey literature using OpenGrey (to 1 October 2013) and ZETOC Conference Proceedings (1993 to 1 October 2013). We placed no restrictions on the language or date of

  20. Formulation of an inhibitor radiopharmaceutical of prostatic antigen of {sup 177}Lu-Glu-Nh-CO-Nh-Lys membrane; Formulacion de un radiofarmaco inhibidor del antigeno prostatico de membrana {sup 177}Lu-Glu-NH-CO-NH-Lys

    Energy Technology Data Exchange (ETDEWEB)

    Ortega S, D.

    2015-07-01

    The prostate specific membrane antigen (PSMA) is a zinc metalloenzyme that is expressed on the cell membrane and highly expressed in prostate cancer. Recently, it has been demonstrated that the peptide sequence Glu-Nh-CO-Nh-Lys inhibit PSMA activity through an electrostatic interaction with the Zn. Several theragnostic radiopharmaceuticals with base in {sup 177}Lu have been developed for radiotherapy of specific molecular targets because gamma and beta emissions of the radionuclide (β = 0.498 MeV and γ= 0.133 MeV). However, there is currently no label a formulation for preparing a radiopharmaceutical of {sup 177}Lu-Glu-Nh-CO-Nh-Lys useful treatment of prostate cancer. The aim of this research was to optimize and document the process of production of the radiopharmaceutical {sup 177}Lu-Glu-Nh-CO-Nh-Lys for sanitary registration application before the Comision Federal para la Proteccion contra Riesgos Sanitarios (COFEPRIS). The optimization of the production process was assessed a factorial design of three variables with mixed levels (3 x 3 x 2) where the dependent variable is the radiochemical purity, the analytical method was validated by UV-Vis spectrophotometry. Next, process validation was carried out by labeling 3 lots of the optimized formulation of the radiopharmaceutical (5.55 GBq (2.16 μg) of {sup 177}LuCl{sub 3}, 90 mg peptide PSMA, 50 mg ascorbic acid and 150 μL of acetate buffer 1 M ph 5), long-term stability was performed by high resolution liquid chromatography) to determine its useful shelf life. 3 validation batches were prepared under protocols of Good Manufacturing Practice (GMP) in the Production Plant of Radiopharmaceuticals of the Instituto Nacional de Investigaciones Nucleares (ININ), meet specifications preset by obtaining a sterile and free development of bacterial endotoxin yields of labeled 100% and which retains its quality characteristics radiochemical purity greater than 90% for at least 15 days. (Author)

  1. Multimodality imaging in apical hypertrophic cardiomyopathy

    Science.gov (United States)

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

    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 first-line 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. PMID:25276293

  2. 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.

  3. 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.

  4. Chloroplast-derived vaccine antigens confer dual immunity against cholera and malaria by oral or injectable delivery.

    Science.gov (United States)

    Davoodi-Semiromi, Abdoreza; Schreiber, Melissa; Nalapalli, Samson; Verma, Dheeraj; Singh, Nameirakpam D; Banks, Robert K; Chakrabarti, Debopam; Daniell, Henry

    2010-02-01

    Cholera and malaria are major diseases causing high mortality. The only licensed cholera vaccine is expensive; immunity is lost in children within 3 years and adults are not fully protected. No vaccine is yet available for malaria. Therefore, in this study, the cholera toxin-B subunit (CTB) of Vibrio cholerae fused to malarial vaccine antigens apical membrane antigen-1 (AMA1) and merozoite surface protein-1 (MSP1) was expressed in lettuce and tobacco chloroplasts. Southern blot analysis confirmed homoplasmy and stable integration of transgenes. CTB-AMA1 and CTB-MSP1 fusion proteins accumulated up to 13.17% and 10.11% (total soluble protein, TSP) in tobacco and up to 7.3% and 6.1% (TSP) in lettuce, respectively. Nine groups of mice (n = 10/group) were immunized subcutaneously (SQV) or orally (ORV) with purified antigens or transplastomic tobacco leaves. Significant levels of antigen-specific antibody titres of immunized mice completely inhibited proliferation of the malarial parasite and cross-reacted with the native parasite proteins in immunoblots and immunofluorescence studies. Protection against cholera toxin challenge in both ORV (100%) and SQV (89%) mice correlated with CTB-specific titres of intestinal, serum IgA and IgG1 in ORV and only IgG1 in SQV mice, but no other immunoglobulin. Increasing numbers of interleukin-10(+) T cell but not Foxp3(+) regulatory T cells, suppression of interferon-gamma and absence of interleukin-17 were observed in protected mice, suggesting that immunity is conferred via the Tr1/Th2 immune response. Dual immunity against two major infectious diseases provided by chloroplast-derived vaccine antigens for long-term (>300 days, 50% of mouse life span) offers a realistic platform for low cost vaccines and insight into mucosal and systemic immunity.

  5. Does Increased Expression of the Plasma Membrane Calcium-ATPase Isoform 2 Confer Resistance to Apoptosis on Breast Cancer Cells?

    National Research Council Canada - National Science Library

    VanHouten, Joshua N

    2008-01-01

    The plasma membrane calcium ATPase isoform 2 (PMCA2) is highly expressed on the apical membrane of mammary epithelial cells during lactation, and is the predominant pump responsible for calcium transport into milk...

  6. A Phase 2a Randomized Study to Evaluate the Safety and Immunogenicity of the 1790GAHB Generalized Modules for Membrane Antigen Vaccine against Shigella sonnei Administered Intramuscularly to Adults from a Shigellosis-Endemic Country

    Directory of Open Access Journals (Sweden)

    Christina W. Obiero

    2017-12-01

    Full Text Available Shigellosis is a mild-to-severe diarrheal infection, caused by the genus Shigella, and is responsible for significant morbidity and mortality worldwide. We evaluated the safety and immunogenicity of an investigational Shigella sonnei vaccine (1790GAHB based on generalized modules for membrane antigens (GMMA in Kenya, a Shigella-endemic country. This phase 2a, observer-blind, controlled randomized study (NCT02676895 enrolled 74 healthy adults aged 18–45 years, of whom 72 were vaccinated. Participants received, in a 1:1:1 ratio, two vaccinations with the 1790GAHB vaccine at doses of either 1.5/25 μg of O antigen (OAg/protein (group 1.5/25 μg or 5.9/100 μg (group 5.9/100 μg at day (D 1 and D29, or vaccination with a quadrivalent meningococcal vaccine at D1 and tetanus, diphtheria, and acellular pertussis vaccine at D29 (control group. Solicited and unsolicited adverse events (AEs, serious AEs (SAEs, and AEs of special interest (neutropenia and reactive arthritis were collected. Anti-S. sonnei lipopolysaccharide (LPS serum immunoglobulin G (IgG geometric mean concentrations (GMC were evaluated at D1, D29, and D57 and compared to anti-S. sonnei LPS antibody levels in convalescent patients naturally exposed to S. sonnei. The percentages of participants with seroresponse were also calculated. The most frequently reported solicited local and systemic AEs across all groups were pain and headache, respectively. Only one case of severe systemic reaction was reported (severe headache after first vaccination in group 5.9/100 μg. Seven and three episodes of neutropenia, assessed as probably or possibly related to vaccination respectively, were reported in the investigational and control groups, respectively. No other SAEs were reported. Despite very high baseline anti-S. sonnei LPS serum IgG levels, the 1790GAHB vaccine induced robust antibody responses. At D29, GMC increased 2.10- and 4.43-fold from baseline in groups 1.5/25 and 5.9/100

  7. Seroprevalence of Antibodies against Plasmodium falciparum Sporozoite Antigens as Predictive Disease Transmission Markers in an Area of Ghana with Seasonal Malaria Transmission.

    Directory of Open Access Journals (Sweden)

    Kwadwo A Kusi

    Full Text Available As an increasing number of malaria-endemic countries approach the disease elimination phase, sustenance of control efforts and effective monitoring are necessary to ensure success. Mathematical models that estimate anti-parasite antibody seroconversion rates are gaining relevance as more sensitive transmission intensity estimation tools. Models however estimate yearly seroconversion and seroreversion rates and usually predict long term changes in transmission, occurring years before the time of sampling. Another challenge is the identification of appropriate antigen targets since specific antibody levels must directly reflect changes in transmission patterns. We therefore investigated the potential of antibodies to sporozoite and blood stage antigens for detecting short term differences in malaria transmission in two communities in Northern Ghana with marked, seasonal transmission.Cross-sectional surveys were conducted during the rainy and dry seasons in two communities, one in close proximity to an irrigation dam and the other at least 20 Km away from the dam. Antibodies against the sporozoite-specific antigens circumsporozoite protein (CSP and Cell traversal for ookinetes and sporozoites (CelTOS and the classical blood stage antigen apical membrane antigen 1 (AMA1 were measured by indirect ELISA. Antibody levels and seroprevalence were compared between surveys and between study communities. Antibody seroprevalence data were fitted to a modified reversible catalytic model to estimate the seroconversion and seroreversion rates.Changes in sporozoite-specific antibody levels and seroprevalence directly reflected differences in parasite prevalence between the rainy and dry seasons and hence the extent of malaria transmission. Seroconversion rate estimates from modelled seroprevalence data did not however support the above observation.The data confirms the potential utility of sporozoite-specific antigens as useful markers for monitoring short term

  8. An apical granuloma with epithelial integument.

    Science.gov (United States)

    Nair, P N; Schmid-Meier, E

    1986-12-01

    A chronic periapicalesion--apical granuloma--of the first left maxillary molar, which had rarefied the bony plate and grown into the maxillary sinus, was observed in a 64-year-old white male patient. Light microscopic study of the epon-embedded specimen showed a lesion that had been infiltrated predominantly by mononuclear cells and that had isolated neutrophil-dominated foci. The lesion contained extensive networks and arcades of proliferating epithelium. Unlike the classic granulomas of the periapex, which are generally delimited by a well-developed connective tissue capsule, this particular lesion was lined with epithelial tissue consisting of ciliated columnar and stratified squamous components. While the former seemed to be an extension of the sinus epithelium, the latter appeared to be an outgrowth of the proliferating epithelium encountered within the lesion.

  9. Apical extrusion of debris using reciprocating files and rotary ...

    African Journals Online (AJOL)

    Procedure: Sixty extracted human mandibular premolars were used. The root canals were instrumented using reciprocating (WaveOne, Reciproc, SafeSider) or rotary ... and cross‑sections, and kinematics, and this situation may influence the amount of apically extruded debris through the apical foramen.[15]. The aim of this ...

  10. Association of ST elevation with apical aneurysm in hypertrophic cardiomyopathy

    Directory of Open Access Journals (Sweden)

    Ozcan Ozeke

    2015-09-01

    Conclusions: Clinicians and specifically echocardiographers must pay special attention on the electrocardiography to correctly detect the frequently overlooked apical aneurysm in HCM patients, and should be careful for apical aneurysm particularly in the presence of STE in V4-6 derivations.

  11. Establishing Apical Patency and its Effect on Endodontic Outcomes

    Science.gov (United States)

    2012-06-01

    accurate prognosis and improved treatment planning related to establishing apical patency. The preliminary results of this interim analysis indicate there...canal space and periodontal ligament. Establishing apical patency is controversial with only 50% of dental programs in the United States teaching the...concept. Both sides can cite articles favoring their position. However, the clinical impact of establishing patency on endodontic treatment outcome

  12. Apical Shell Sculpture of some African Freshwater Limpets ...

    African Journals Online (AJOL)

    A Scanning Electron Microscope has been used to describe in detail the apical shell sculpture of sevenAfrican species (three genera) of freshwater limpet snails (Ancylidae). The apical sculpture of Ancylus fluviatilis (? syn. strigatus and ? brondeli) is striate, but quite different in basic pattern from the other three striate ...

  13. Apical Hypertrophic Cardiomyopathy in Association with PulmonaryArtery Hypertension

    Directory of Open Access Journals (Sweden)

    Mehdi Peighambari

    2012-09-01

    Full Text Available Apical Hypertrophic Cardiomyopathy is an uncommon condition constituting 1% -2% of the cases with Hypertrophic Cardiomyopathy (HCM diagnosis. We interestingly report two patients with apical hypertrophic cardiomyopathy in association with significant pulmonary artery hypertension without any other underlying reason for pulmonary hypertension. The patients were assessed by echocardiography, cardiac catheterization and pulmonary function parameters study.

  14. Apical extrusion of debris using reciprocating files and rotary ...

    African Journals Online (AJOL)

    ... the other groups, with the exception of the Typhoon group (P < 0.05). Conclusion: According to our study, all systems caused apical debris extrusion. However, the Reciproc group was associated with less debris extrusion when compared to the other groups. Key words: Apical extrusion, endodontics, single file systems ...

  15. Diagnosis of apical hypertrophic cardiomyopathy: T-wave inversion and relative but not absolute apical left ventricular hypertrophy.

    Science.gov (United States)

    Flett, Andrew S; Maestrini, Viviana; Milliken, Don; Fontana, Mariana; Treibel, Thomas A; Harb, Rami; Sado, Daniel M; Quarta, Giovanni; Herrey, Anna; Sneddon, James; Elliott, Perry; McKenna, William; Moon, James C

    2015-03-15

    Diagnosis of apical HCM utilizes conventional wall thickness criteria. The normal left ventricular wall thins towards the apex such that normal values are lower in the apical versus the basal segments. The impact of this on the diagnosis of apical hypertrophic cardiomyopathy has not been evaluated. We performed a retrospective review of 2662 consecutive CMR referrals, of which 75 patients were identified in whom there was abnormal T-wave inversion on ECG and a clinical suspicion of hypertrophic cardiomyopathy. These were retrospectively analyzed for imaging features consistent with cardiomyopathy, specifically: relative apical hypertrophy, left atrial dilatation, scar, apical cavity obliteration or apical aneurysm. For comparison, the same evaluation was performed in 60 healthy volunteers and 50 hypertensive patients. Of the 75 patients, 48 met conventional HCM diagnostic criteria and went on to act as another comparator group. Twenty-seven did not meet criteria for HCM and of these 5 had no relative apical hypertrophy and were not analyzed further. The remaining 22 patients had relative apical thickening with an apical:basal wall thickness ratio >1 and a higher prevalence of features consistent with a cardiomyopathy than in the control groups with 54% having 2 or more of the 4 features. No individual in the healthy volunteer group had more than one feature and no hypertension patient had more than 2. A cohort of individuals exist with T wave inversion, relative apical hypertrophy and additional imaging features of HCM suggesting an apical HCM phenotype not captured by existing diagnostic criteria. Copyright © 2015 Elsevier Ireland Ltd. All rights reserved.

  16. Carcinoma-associated antigens

    International Nuclear Information System (INIS)

    Bartorelli, A.; Accinni, R.

    1981-01-01

    This invention relates to novel antigens associated with breast carcinoma, anti-sera specific to said antigens, 125 I-labeled forms of said antigens and methods of detecting said antigens in serum or plasma. The invention also relates to a diagnostic kit containing standardised antigens or antisera or marked forms thereof for the detection of said antigens in human blood, serum or plasma. (author)

  17. Efficacy of Biodentine as an Apical Plug in Nonvital Permanent Teeth with Open Apices: An In Vitro Study

    Directory of Open Access Journals (Sweden)

    Mehmet Bani

    2015-01-01

    Full Text Available The aim of this study was to evaluate the apical microleakage of Biodentine and MTA orthograde apical plugs and to compare the effect of thickness of these biomaterials on their sealing ability. A total of eighty maxillary anterior teeth were used. The apices were removed by cutting with a diamond disc (Jota, Germany 2 mm from the apical root end in an attempt to standardize the working length of all specimens to 15 ± 1 mm. Both materials were placed in 1–4 mm thickness as apical plugs root canal. Root canal leakage was evaluated by the fluid filtration technique. One-way ANOVA was used in order to determine normality of dispersal distribution of parameters; thereafter, results were analyzed by Kolmogorov-Smirnov test. Overall, between microleakage values of MTA and Biodentine regardless of apical plug thickness, no difference was observed. In terms of plug thickness, a statistically significant difference was observed between the subgroups of MTA and Biodentine (p<0.05. The apical sealing ability of Biodentine was comparable to MTA at any apical plug thickness.

  18. The V0-ATPase mediates apical secretion of exosomes containing Hedgehog-related proteins in Caenorhabditis elegans

    Science.gov (United States)

    Liégeois, Samuel; Benedetto, Alexandre; Garnier, Jean-Marie; Schwab, Yannick; Labouesse, Michel

    2006-01-01

    Polarized intracellular trafficking in epithelia is critical in development, immunity, and physiology to deliver morphogens, defensins, or ion pumps to the appropriate membrane domain. The mechanisms that control apical trafficking remain poorly defined. Using Caenorhabditis elegans, we characterize a novel apical secretion pathway involving multivesicularbodies and the release of exosomes at the apical plasma membrane. By means of two different genetic approaches, we show that the membrane-bound V0 sector of the vacuolar H+-ATPase (V-ATPase) acts in this pathway, independent of its contribution to the V-ATPase proton pump activity. Specifically, we identified mutations in the V0 “a” subunit VHA-5 that affect either the V0-specific function or the V0+V1 function of the V-ATPase. These mutations allowed us to establish that the V0 sector mediates secretion of Hedgehog-related proteins. Our data raise the possibility that the V0 sector mediates exosome and morphogen release in mammals. PMID:16785323

  19. Alpha-enolase on apical surface of renal tubular epithelial cells serves as a calcium oxalate crystal receptor

    Science.gov (United States)

    Fong-Ngern, Kedsarin; Thongboonkerd, Visith

    2016-10-01

    To search for a strategy to prevent kidney stone formation/recurrence, this study addressed the role of α-enolase on apical membrane of renal tubular cells in mediating calcium oxalate monohydrate (COM) crystal adhesion. Its presence on apical membrane and in COM crystal-bound fraction was confirmed by Western blotting and immunofluorescence staining. Pretreating MDCK cells with anti-α-enolase antibody, not isotype-controlled IgG, dramatically reduced cell-crystal adhesion. Immunofluorescence staining also confirmed the direct binding of purified α-enolase to COM crystals at {121} > {100} > {010} crystal faces. Coating COM crystals with urinary proteins diminished the crystal binding capacity to cells and purified α-enolase. Moreover, α-enolase selectively bound to COM, not other crystals. Chemico-protein interactions analysis revealed that α-enolase interacted directly with Ca2+ and Mg2+. Incubating the cells with Mg2+ prior to cell-crystal adhesion assay significantly reduced crystal binding on the cell surface, whereas preincubation with EDTA, a divalent cation chelator, completely abolished Mg2+ effect, indicating that COM and Mg2+ competitively bind to α-enolase. Taken together, we successfully confirmed the role of α-enolase as a COM crystal receptor to mediate COM crystal adhesion at apical membrane of renal tubular cells. It may also serve as a target for stone prevention by blocking cell-crystal adhesion and stone nidus formation.

  20. Healing of apical periodontitis through modern endodontic retreatment techniques.

    Science.gov (United States)

    Ray, Jarom J; Kirkpatrick, Timothy C

    2013-01-01

    The presence of apical periodontitis in teeth which have undergone initial root canal treatment is largely attributed to bacteria residing in or invading from the apical root canal space. Bacteria-associated apical periodontitis will not heal spontaneously, nor will systemic antibiotics eradicate the infection. Only endodontic retreatment, endodontic surgery, or extraction will control the bacterial etiology. Modern retreatment is an effective means of addressing apical periodontitis. A mandibular premolar with apical periodontitis, apical root resorption, and overfilled gutta percha was retreated with post removal, retrieval of gutta percha from beyond the apex, ultrasonic irrigation and disinfection, and placement of a collagen internal matrix to facilitate a well-controlled MTA apical fill. The magnification and illumination imparted by the operating microscope was integral to achievement of treatment objectives. The patient's symptoms were resolved and complete osseous healing occurred. During treatment planning, clinicians should consider the capability of modern endodontic techniques to overcome technical challenges, often allowing the natural dentition to be preserved and restored to function days after retreatment.

  1. ANTIGENIC PROMOTION

    Science.gov (United States)

    Wu, Chin-Yu; Cinader, Bernard

    1971-01-01

    Rabbits were immunized with p-azobenzene arsonic acid derivatives of human serum albumin (HA-As) or of dissociated keyhole limpet hemocyanin. The IgM response to the hapten was evaluated in terms of the number of hapten-specific plaque-forming cells in the lymph node draining the injection site. In some experiments, antibody was measured by agglutination of tanned and sensitized erythrocytes. The hapten response of animals immunized with HA-As was increased (promoting effect) when the animals were injected with one of several structurally unrelated macromolecules: keyhole limpet hemocyanin (KLH), horse spleen ferritin (HSF), lysozyme (Lys), alum-precipitated human gamma globulin (alum-precipitated HGG). Different macromolecules differed in the magnitude of the promoting effect they induced, e.g., promotion by the associated form of KLH was greater than that by the dissociated form; alum-precipitated HGG was a better promoter than was soluble HGG. The relative magnitude of promotion by different macromolecules (associated vs. dissociated KLH, alum-precipitated vs. soluble HGG) correlated with the relative magnitude of the carrier effect, as judged by the hapten response induced by p-azobenzene arsonic acid conjugated to various proteins. Promotion was detected by agglutination assay of circulating antibody, by plaque assay of cells from the popliteal lymph node draining the site of preinjection, but not by plaque assay of cells from the contralateral lymph node. Promotion was dependent on the dose of the promoting macromolecule and on the dose of the hapten-protein conjugate. It was not observed in animals tolerant to the promoting macromolecule. Inhibition (i.e. antigenic competition), rather than promotion, was observed upon a secondary response to the preinjected macromolecule or when the hapten-protein conjugate was incorporated in Freund's adjuvant. PMID:15776570

  2. 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...

  3. Mineral trioxide aggregate apical plugs in teeth with open apical foramina: a retrospective analysis of treatment outcome.

    Science.gov (United States)

    Mente, Johannes; Hage, Nathalie; Pfefferle, Thorsten; Koch, Martin Jean; Dreyhaupt, Jens; Staehle, Hans Joerg; Friedman, Shimon

    2009-10-01

    Teeth with open apical foramina present a challenge during root canal treatment, and little is known about the clinical outcome of treatment in such teeth. This retrospective study assessed healing of teeth with open apices managed by the placement of mineral trioxide aggregate apical plugs. Seventy-two patients with 78 teeth with apical resorption or excessive apical enlargement, treated between 2000 and 2006, were contacted for follow-up examination 12 to 68 months after treatment (median 30.9 months). Treatments were provided by supervised undergraduate students (27%), general dentists (32%), or dentists who had focused on endodontics (41%). The outcome based on clinical and radiographic criteria was assessed by calibrated examiners and dichotomized as "healed" or "disease." Of 56 teeth examined (72% recall), 84% were healed. Teeth without or with preoperative periapical radiolucency had a healed rate of 100% and 78%, respectively. None of the variables analyzed had a significant effect on the outcome. The results supported the management of open apical foramina with mineral trioxide aggregate apical plugs.

  4. Efficacy of Biodentine as an Apical Plug in Nonvital Permanent Teeth with Open Apices: An In Vitro Study.

    Science.gov (United States)

    Bani, Mehmet; Sungurtekin-Ekçi, Elif; Odabaş, Mesut Enes

    2015-01-01

    The aim of this study was to evaluate the apical microleakage of Biodentine and MTA orthograde apical plugs and to compare the effect of thickness of these biomaterials on their sealing ability. A total of eighty maxillary anterior teeth were used. The apices were removed by cutting with a diamond disc (Jota, Germany) 2 mm from the apical root end in an attempt to standardize the working length of all specimens to 15 ± 1 mm. Both materials were placed in 1-4 mm thickness as apical plugs root canal. Root canal leakage was evaluated by the fluid filtration technique. One-way ANOVA was used in order to determine normality of dispersal distribution of parameters; thereafter, results were analyzed by Kolmogorov-Smirnov test. Overall, between microleakage values of MTA and Biodentine regardless of apical plug thickness, no difference was observed. In terms of plug thickness, a statistically significant difference was observed between the subgroups of MTA and Biodentine (p Biodentine was comparable to MTA at any apical plug thickness.

  5. Polyclonal antibodies for the detection of Trypanosoma cruzi circulating antigens.

    Directory of Open Access Journals (Sweden)

    Edith S Málaga-Machaca

    2017-11-01

    Full Text Available Detection of Trypanosoma cruzi antigens in clinical samples is considered an important diagnostic tool for Chagas disease. The production and use of polyclonal antibodies may contribute to an increase in the sensitivity of immunodiagnosis of Chagas disease.Polyclonal antibodies were raised in alpacas, rabbits, and hens immunized with trypomastigote excreted-secreted antigen, membrane proteins, trypomastigote lysate antigen and recombinant 1F8 to produce polyclonal antibodies. Western blot analysis was performed to determine specificity of the developed antibodies. An antigen capture ELISA of circulating antigens in serum, plasma and urine samples was developed using IgY polyclonal antibodies against T. cruzi membrane antigens (capture antibody and IgG from alpaca raised against TESA. A total of 33 serum, 23 plasma and 9 urine samples were analyzed using the developed test. Among serum samples, compared to serology, the antigen capture ELISA tested positive in 55% of samples. All plasma samples from serology positive subjects were positive in the antigen capture ELISA. All urine positive samples had corresponding plasma samples that were also positive when tested by the antigen capture ELISA.Polyclonal antibodies are useful for detection of circulating antigens in both the plasma and urine of infected individuals. Detection of antigens is direct evidence of the presence of the parasite, and could be a better surrogate of current infection status.

  6. 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

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

    Science.gov (United States)

    Kabasakal, Levent; AbuQbeitah, Mohammad; Aygün, Aslan; Yeyin, Nami; Ocak, Meltem; Demirci, Emre; Toklu, Turkay

    2015-12-01

    (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 (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 (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 method. The dose-limiting organ seems to be the parotid glands rather than kidneys and bone marrow. The lesion radiation doses are within acceptable ranges; however, there is a substantial individual variance so patient dosimetry seems to be mandatory.

  8. Phenotypic overlap in hypertrophic cardiomyopathy: apical hypertrophy, midventricular obstruction, and apical aneurysm.

    Science.gov (United States)

    Minami, Yuichiro; Haruki, Shintaro; Hagiwara, Nobuhisa

    2014-12-01

    Within the diverse phenotypic spectrum of hypertrophic cardiomyopathy (HCM), subgroups of patients with apical hypertrophy (APH), midventricular obstruction (MVO), and apical aneurysm (APA) have emerged. While previous studies have suggested the existence of considerable overlap between APH, MVO, and APA, there are still many unanswered questions. Therefore, we attempted to clarify the relationship of the above three phenotypes of HCM with respect to prevalence, overlap, and outcomes. Among the 544 study HCM patients (mean follow-up period: 11.6±7.4 years), 170 with APH (31.3%), 51 with MVO (9.4%), and 24 with APA (4.4%) were examined. There was phenotypic overlap between APH and MVO in 17 patients, APH and APA in 14 patients, and MVO and APA in 14 patients. Furthermore, a combination of APH, MVO, and APA was observed in eight patients. Detailed analysis of the relationship between overlapping phenotypes and the prognosis showed that APA patients without a history of APH had an extremely poor outcome (probability of the combined endpoint of sudden death and potentially lethal arrhythmic events ≥50%). Conversely, APH patients without MVO had a strikingly good outcome (probability of the combined endpoint <5%). Other patients had an intermediate outcome (probability of the combined endpoint 10-40%). Our results suggest that overlap between these three forms of HCM is substantial, and that detailed classification of the overlapping phenotypes is clinically meaningful. Copyright © 2014 Japanese College of Cardiology. Published by Elsevier Ltd. All rights reserved.

  9. Pathogenesis of apical periodontal cysts: guidelines for diagnosis in palaeopathology

    OpenAIRE

    Dias, G. J.; Prasad, K.; Santos, A. L.

    2007-01-01

    Apical periodontal cysts are benign lesions developing in relation to the apices of non-vital teeth due to inflammatory response from the infective pulp. These are epithelium-lined bony cavities containing fluid. Despite being widely reported in medical/dental literature, this common condition is poorly diagnosed and documented in the archaeological literature. We aim to clarify the correct terminology, demonstrate bony manifestations at different stages of pathogenesis of chronic periapical ...

  10. Hypertrophic cardiomyopathy with mid-ventricular obstruction and apical aneurysm

    Directory of Open Access Journals (Sweden)

    N.D. Oryshchyn

    2016-11-01

    Full Text Available A case report of apical left ventricular aneurysm in patient with hypertrophic cardiomyopathy with mid-ventricular obstruction (diagnosis and surgical treatment is presented. We revealed apical aneurysm and mid-ventricular obstruction during echocardiography and specified anatomical characteristics of aneurysm during computer tomography. There was no evidence of obstructive coronary artery disease during coronary angiography. Taking into consideration multiple cerebral infarcts, aneurysm resection and left ventricular plastics was performed. Electronic microscopy of myocardium confirmed the diagnosis of hypertrophic cardiomyopathy.

  11. Apical Negative Pressure irrigation presents tissue compatibility in immature teeth

    Directory of Open Access Journals (Sweden)

    Carolina Maschietto Pucinelli

    Full Text Available Abstract Aim: To compare the apical negative pressure irrigation (ANP with conventional irrigation in the teeth of immature dogs with apical periodontitis. Methods: Fifty-two immature pre-molar root canals were randomly assigned into 4 groups: ANP (n=15; conventional irrigation (n=17; healthy teeth (control (n = 10; and teeth with untreated apical periodontitis (control (n=10. After induction of apical periodontitis, teeth were instrumented using EndoVac® (apical negative pressure irrigation or conventional irrigation. The animals were euthanized after 90 days. The sections were stained by HE and analyzed under conventional and fluorescence microscopy. TRAP histoenzymology was also performed. Statistical analyses were performed with the significance level set at 5%. Results: There was difference in the histopathological parameters between ANP and conventional groups (p0.05. However, a lower number of osteoclasts was observed in the ANP group (p<0.05. Conclusion: The EndoVac® irrigation system presented better biological results and more advanced repair process in immature teeth with apical periodontitis than the conventional irrigation system, confirming the hypothesis.

  12. Monocyte chemotactic protein-3: possible involvement in apical periodontitis chemotaxis.

    Science.gov (United States)

    Dezerega, A; Osorio, C; Mardones, J; Mundi, V; Dutzan, N; Franco, M; Gamonal, J; Oyarzún, A; Overall, C M; Hernández, M

    2010-10-01

    To study the expression of monocyte chemotactic protein-3 (MCP-3, also known as chemokine CCL-7) in tissue from apical lesions (AL) and to associate MCP-3 expression with symptomatic or asymptomatic apical periodontitis. To determine the expression of MCP-3 in AL, biopsies obtained during tooth extraction procedures were fixed, subjected to routine processing and diagnosed as apical granuloma (AG) (n = 7) or radicular cyst (RC) (n = 5). As controls, apical periodontal ligament (PDL) specimens from healthy premolars extracted for orthodontics reasons were included (n = 7). All specimens were immunostained for MCP-3 and examined under a light microscope. In addition, homogenates from AL (n = 14) and healthy PDL samples (n = 7) were studied through immunowestern blot. Finally, periapical exudates samples were collected from root canals of teeth having diagnosis of symptomatic (n = 14) and asymptomatic apical periodontitis (n = 14) during routine endodontic treatments and analysed by immunowestern blot and densitometry.   MCP-3 was detected in AG and RC and localized mainly to inflammatory leucocytes, whereas no expression was observed in healthy PDLs. MCP-3 was also detected in periapical exudate, and its levels were significantly higher in symptomatic than in asymptomatic apical periodontitis. MCP-3 was expressed in AL and its levels associated with clinical symptoms. MCP-3 might play a role in disease pathogenesis, possibly by stimulating mononuclear chemotaxis. © 2010 International Endodontic Journal.

  13. 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...

  14. Aquaporin-2 membrane targeting

    DEFF Research Database (Denmark)

    Olesen, Emma T B; Fenton, Robert A

    2017-01-01

    The targeting of the water channel aquaporin-2 (AQP2) to the apical plasma membrane of kidney collecting duct principal cells is regulated mainly by the antidiuretic peptide hormone arginine vasopressin (AVP). This process is of crucial importance for the maintenance of body water homeostasis....... In this brief review we assess the role of cyclic adenosine monophosphate (cAMP) and discuss the emerging concept that type 2 AVP receptor (V2R)-mediated AQP2 trafficking is cAMP-independent. the ability of the kidney to concentrate the urine and thereby maintain body water homeostasis depends on targeting....... For example, 1) stimulation with the nonspecific AC activator forskolin increases AQP2 membrane accumulation in a mouse cortical collecting duct cell line [e.g., Norregaard et al. (16)]; 2) cAMP increases CD water permeability (15); 3) the cAMP-activated protein kinase A (PKA) can phosphorylate AQP2 on its...

  15. CT findings in apical versus basal involvement of pulmonary tuberculosis.

    Science.gov (United States)

    Yoon, Ji Young; Lee, In Jae; Im, Hyoung June; Lee, Kwanseop; Lee, Yul; Bae, Sang Hoon

    2013-01-01

    We aimed to compare clinical features and computed tomography (CT) findings of pulmonary tuberculosis (TB) in lower lobe basal segments and upper lobe apical or apicoposterior segments. We retrospectively reviewed medical records and chest CT scans of 986 adults who were diagnosed with active pulmonary TB. Active pulmonary TB confined to the basal segments was found in 21 patients. Sixty patients had disease localized to the apical or apicoposterior segments only. Clinical features and CT abnormalities of the lung parenchyma, airways, mediastinal and hilar lymph nodes, and pleura were compared between these two groups. A significant difference was observed between two groups in terms of underlying disease prevalence associated with an immunocompromised state (basal, 6/21, 28.6%; apical or apico- posterior, 3/60, 5%; P = 0.008). Chest CT findings, including consolidation (P = 0.0016), lymphadenopathy (P = 0.0297), and pleural effusion (P = 0.008), were more common in basal segment TB than in apical or apicoposterior segment TB. Small nodules were less common in basal segment TB than in apical or apicoposterior segment TB (P = 0.0299). The tree-in-bud sign was the most common CT finding in both basal segment TB (17/21, 81%) and apical or apicoposterior segment TB groups (53/60, 88.3%) (P = 0.4633). Lower lobe basal segment TB was more commonly present with common CT findings of primary pulmonary TB including consolidation, mediastinal and hilar lymphadenopathy, and pleural effusion than apical or apicoposterior segment TB.

  16. Comparison of periapical radiography and limited cone-beam tomography in posterior maxillary teeth referred for apical surgery.

    Science.gov (United States)

    Low, Kenneth M T; Dula, Karl; Bürgin, Walter; von Arx, Thomas

    2008-05-01

    This study compared periapical (PA) radiography and cone-beam tomography (CBT) for preoperative diagnosis in posterior maxillary teeth of consecutive patients referred for possible apical surgery. Images were concurrently analyzed by an oral radiologist and an endodontist to reach consensus in interpretation of the radiographic findings. The final material included 37 premolars and 37 molars with a total of 156 roots. CBT showed significantly more lesions (34%, p radiography. Detecting lesions with PA radiography alone was most difficult in second molars or in roots in close proximity to the maxillary sinus floor. Additional findings were seen significantly more frequently in CBT compared with PA radiography including expansion of lesions into the maxillary sinus (p < 0.001), sinus membrane thickening (p < 0.001), and missed canals (p < 0.05). The present study highlights the advantages of using CBT for preoperative treatment planning in maxillary posterior teeth with apical pathology.

  17. Participation of IAA in transduction of gravistimulus in apical cells of moss protonema

    Science.gov (United States)

    Oksyniuk, U. A.; Khorkavtsiv, O. Y.; Lesniak, Y. I.

    carried out experiments it can be suggested that high concentrations of IAA and 1-NAA result in surplus of IAA cells led, probably, to a destruction of the apical-basal gradient in cells. Our results testify that NPA inhibits the gravitropism stronger than the growth of protonema. The peculiarity of moss protonema is that the growth orientation change is a result of a transference of growth zone in the apical cell dome caused by amyloplasts sedimentation inducing lateral asymmetry of Ca2+ and apical-basal IAA flow what in its turn manifests itself in distribution of IAA and/or Ca2+ channels in apical cell dome plasma membrane ( Schwuchow et al., 2001). The transport of IAA in apical cells, probably, functionally polarizes it and just that polarizing function is dominant in cells with tip growth.

  18. Simple, mild, one-step labelling of proteins with gallium-68 using a tris(hydroxypyridinone) bifunctional chelator: a 68Ga-THP-scFv targeting the prostate-specific membrane antigen.

    Science.gov (United States)

    Nawaz, Saima; Mullen, Gregory E D; Sunassee, Kavitha; Bordoloi, Jayanta; Blower, Philip J; Ballinger, James R

    2017-10-25

    Labelling proteins with gallium-68 using bifunctional chelators is often problematic because of unsuitably harsh labelling conditions such as low pH or high temperature and may entail post-labelling purification. To determine whether tris(hydroxypyridinone) (THP) bifunctional chelators offer a potential solution to this problem, we have evaluated the labelling and biodistribution of a THP conjugate with a new single-chain antibody against the prostate-specific membrane antigen (PSMA), an attractive target for staging prostate cancer (PCa). A single-chain variable fragment (scFv) of J591, a monoclonal antibody that recognises an external epitope of PSMA, was prepared in order to achieve biokinetics matched to the half-life of gallium-68. The scFv, J591c-scFv, was engineered with a C-terminal cysteine. J591c-scFv was produced in HEK293T cells and purified by size-exclusion chromatography. A maleimide THP derivative (THP-mal) was coupled site-specifically to the C-terminal cysteine residue. The THP-mal-J591c-scFv conjugate was labelled with ammonium acetate-buffered gallium-68 from a 68 Ge/ 68 Ga generator at room temperature and neutral pH. The labelled conjugate was evaluated in the PCa cell line DU145 and its PSMA-overexpressing variant in vitro and xenografted in SCID mice. J591c-scFv was produced in yields of 4-6 mg/l culture supernatant and efficiently coupled with the THP-mal bifunctional chelator. Labelling yields > 95% were achieved at room temperature following incubation of 5 μg conjugate with gallium-68 for 5 min without post-labelling purification. 68 Ga-THP-mal-J591c-scFv was stable in serum and showed selective binding to the DU145-PSMA cell line, allowing an IC50 value of 31.5 nM to be determined for unmodified J591c-scFv. Serial PET/CT imaging showed rapid, specific tumour uptake and clearance via renal elimination. Accumulation in DU145-PSMA xenografts at 90 min post-injection was 5.4 ± 0.5%ID/g compared with 0.5 ± 0.2%ID/g in DU145

  19. Apical Revascularization after Delayed Tooth Replantation: An Unusual Case

    Directory of Open Access Journals (Sweden)

    Marília Pacífico Lucisano

    2016-01-01

    Full Text Available The aim of this paper is to present the clinical and radiological outcome of the treatment involving a delayed tooth replantation after an avulsed immature permanent incisor, with a follow-up of 1 year and 6 months. An 8-year-old boy was referred after dental trauma that occurred on the previous day. The permanent maxillary right central incisor (tooth 11 had been avulsed. The tooth was hand-held during endodontic therapy and an intracanal medication application with calcium hydroxide-based paste was performed. An apical plug with mineral trioxide aggregate (MTA was introduced into the apical portion of the canal. When the avulsed tooth was replanted with digital pressure, a blood clot had formed within the socket, which moved the MTA apical plug about 2 mm inside of the root canal. These procedures developed apical revascularization, which promoted a successful endodontic outcome, evidenced by apical closure, slight increase in root length, and absence of signs of external root resorption, during a follow-up of 1 year and 6 months.

  20. [Histopathology and etiopathogenesis of chronic apical periodontitis--periapical granuloma].

    Science.gov (United States)

    Kovác, J; Kovác, D

    2011-06-01

    Periapical lesions are among the most frequently diagnosed apical odontogenic pathologies in human teeth. The condition is generally described as apical periodontitis. 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 to the periapical tissue. 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. There are also factors located within the inflamed periapical tissue that can interfere with post-treatment healing of the lesion. The purpose of this article is to provide a comprehensive overview of the etiopathogenesis of apical periodontitis and causes of failed endodontic treatment. This study presents a histopathological analysis through optical microscopy of periapical lesions, commonly referred to as solid dental or periapical granuloma.

  1. [Mineral trioxide aggregate (MTA) a success story in apical surgery].

    Science.gov (United States)

    von Arx, Thomas

    2016-01-01

    The objective of apical surgery is to retain teeth with persistent apical pathosis following orthograde root canal treatment if endodontic non-surgical revision is difficult or associated with risks, or is even declined by the patient. Since the most frequent cause of recurrent apical disease is bacterial reinfection from the (remaining) root canal system, the bacteria-tight root-end filling is the most important step in apical surgery. In the early 1990s, mineral trioxide aggregate (MTA) was developed at the Loma Linda University in California/USA. Preclinical studies clearly showed that MTA has a high sealing capability, a good material stability and an excellent biocompatbility. Multiple experimental studies in animals highlighted the mild tissue reactions observed adjacent to this material. Furthermore, histological analysis of the periapical regions demonstrated a frequent deposition of new cementum not only onto the resection plane (cut dentinal surface), but also directly onto MTA. For these reasons, MTA is considered a bioactive material. In 1997 MTA was cleared for clinical use in patients. Multiple prospective clinical and randomized studies have documented high and constant success rates of MTA-treated teeth in apical surgery. A recently published longitudinal study showed that MTA-treated teeth remained stable over five years; hence the high healed rates documented after one year are maintained during long-term observation.

  2. 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...... in the soluble fraction, suggesting a basolateral secretion into the intercellular space, and both this accumulation and the release to the medium was prevented by culture at 20 degrees C. The specific radioactivity of apo A-1 and apo B-48 released to the medium was significantly higher than...... 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...

  3. The Apical Localization of Na+, K+-ATPase in Cultured Human Retinal Pigment Epithelial Cells Depends on Expression of the β2 Subunit.

    Science.gov (United States)

    Lobato-Álvarez, Jorge A; Roldán, María L; López-Murillo, Teresa Del Carmen; González-Ramírez, Ricardo; Bonilla-Delgado, José; Shoshani, Liora

    2016-01-01

    Na + , K + -ATPase, or the Na + pump, is a key component in the maintenance of the epithelial phenotype. In most epithelia, the pump is located in the basolateral domain. Studies from our laboratory have shown that the β 1 subunit of Na + , K + -ATPase plays an important role in this mechanism because homotypic β 1 -β 1 interactions between neighboring cells stabilize the pump in the lateral membrane. However, in the retinal pigment epithelium (RPE), the Na + pump is located in the apical domain. The mechanism of polarization in this epithelium is unclear. We hypothesized that the apical polarization of the pump in RPE cells depends on the expression of its β 2 subunit. ARPE-19 cells cultured for up to 8 weeks on inserts did not polarize, and Na + , K + -ATPase was expressed in the basolateral membrane. In the presence of insulin, transferrin and selenic acid (ITS), ARPE-19 cells cultured for 4 weeks acquired an RPE phenotype, and the Na + pump was visible in the apical domain. Under these conditions, Western blot analysis was employed to detect the β 2 isoform and immunofluorescence analysis revealed an apparent apical distribution of the β 2 subunit. qPCR results showed a time-dependent increase in the level of β 2 isoform mRNA, suggesting regulation at the transcriptional level. Moreover, silencing the expression of the β 2 isoform in ARPE-19 cells resulted in a decrease in the apical localization of the pump, as assessed by the mislocalization of the α 2 subunit in that domain. Our results demonstrate that the apical polarization of Na + , K + -ATPase in RPE cells depends on the expression of the β 2 subunit.

  4. 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.

  5. Spontaneous coronary artery dissection associated with apical hypertrophic cardiomyopathy

    International Nuclear Information System (INIS)

    Tuncer, M.; Gumrukcuoglu, H.A.; Ekim, H.; Gunes, Y.; Simsek, H.

    2010-01-01

    Apical hypertrophic cardiomyopathy (HCM) is a relatively uncommon inherited disease. Spontaneous coronary artery dissection (SCAD) is also uncommonly observed, which often occurs in pregnant or post partum women but is rare in men. This report describes a 38 years old man with apical hypertrophic cardiomyopathy who developed SCAD leading to acute inferior myocardial infarction. After emergent appendectomy operation at another hospital, he was immediately transferred to the Cardiology Department of our hospital due to acute myocardial infarction. He emergently underwent coronary angiography which showed a long dissection involving the right coronary. He underwent an emergent CABG with cardiopulmonary bypass. Postoperative recovery was uneventful and he was discharged. According to our knowledge, no case of spontaneous coronary artery dissection associated with apical hypertrophic cardiomyopathy unrelated to postpartum period or oral contraceptive use has been reported so far. (author)

  6. Mineral trioxide aggregate as apical plug in teeth with necrotic pulp and immature apices: a 10-year case series.

    Science.gov (United States)

    Pace, Riccardo; Giuliani, Valentina; Nieri, Michele; Di Nasso, Luca; Pagavino, Gabriella

    2014-08-01

    This 10-year study evaluated the clinical and radiologic outcomes of teeth with necrotic pulp, immature apices, and periapical lesions treated with the mineral trioxide aggregate (MTA) apical plug technique. Seventeen single-rooted immature teeth with necrotic pulp and periapical lesion from 17 patients treated between January 2001 and December 2001 were included in this study. Apical obturation on all teeth included in the study was completed in 2 visits: first using calcium hydroxide as an interappointment intracanal medication and a second visit for the creation of the artificial apical barrier with MTA. The outcome, based on clinical and radiographic criteria, was assessed by 2 calibrated investigators using the periapical index (PAI). The Friedman test was used to verify the differences between baseline and the 1-, 5-, and 10-year PAI scores. Of the 17 patients treated, 1 patient dropped out at 5 years. At the 10-year follow-up, 15 teeth were healed (PAI ≤2), and 1 tooth had been extracted because of the presence of a longitudinal root fracture. The PAI score exhibited a significant decrease between baseline and 1 year and between 1 and 5 years. The difference between 5 and 10 years was not significant. The apical plug with MTA was a successful and effective technique for long-term management of this group of teeth with necrotic pulps with immature root development and periapical lesions. Copyright © 2014 American Association of Endodontists. Published by Elsevier Inc. All rights reserved.

  7. Phosphate depletion in opossum kidney cells: apical but not basolateral or transepithelial adaptions of Pi transport.

    Science.gov (United States)

    Barac-Nieto, M; Alfred, M; Spitzer, A

    2001-01-01

    Monolayers of opossum kidney (OK) cells are widely used as models for the renal proximal tubule. OK cells adapt to phosphate (Pi) depletion by increasing their capacity for apical and basolateral Na+-dependent Pi uptake. Because NMR-visible cell Pi was found to be decreased in Pi-deprived kidney cells, we suggested that up-regulation of basolateral Pi efflux also occurs during adaptation to Pi deprivation [American Journal of Physiol 1994;267:C915-919]. In order to test this hypothesis, we measured the cell Pi pool, basolateral Pi efflux and transepithelial Pi fluxes in OK cells grown on permeable plastic filters, exposed overnight to solutions containing either 0.5 mM (deprived) or 2.0 mM (replete) Pi or 32Pi. Following steady state or acute loading with 32Pi, the specific activity (SA) of cell Pi, the cell Pi pool and the basolateral efflux of 32Pi were measured. In the steady state, a 2-fold increase in Pi uptake sustained the intracellular Pi pool at 85% of the control level (30 +/- 5 nmol/mg) in spite of a decrease in extracellular Pi from 2 to 0.5 mM. When the extracellular Pi was acutely (1 h) reduced to 0.1 mM, the cell Pi pool decreased (to 3 +/- 1 nmol/mg) both in cells previously adapted overnight to either 0.5 or to 2 mM Pi (p >0.3). The rates of absolute and fractional basolateral washout of cell 32Pi after 1 h loading with 0.1 mM 32Pi were similar in cells adapted to 0.5 compared to 2 mM Pi. This indicates that Pi depletion did not affect the effective permeability of the basolateral membranes to Pi. Adaptation for 16 h to 0.5 compared to 2 mM Pi did not alter the rate of net transepithelial transport of 0.1 mM Pi from the apical to the basal compartment but reduced (p OK cells grown on plastic support there are no adaptive increases in either basolateral Pi efflux, or in transcellular and paracellular Pi transport, in response to Pi depletion. Adaptations are limited to increases in apical and basolateral sodium-dependent Pi uptakes that can maintain

  8. Membrane fusion triggers rapid degradation of two gamete-specific, fusion-essential proteins in a membrane block to polygamy in Chlamydomonas

    OpenAIRE

    Liu, Yanjie; Misamore, Michael J.; Snell, William J.

    2010-01-01

    The plasma membranes of gametes are specialized for fusion, yet, once fusion occurs, in many organisms the new zygote becomes incapable of further membrane fusion reactions. The molecular mechanisms that underlie this loss of fusion capacity (block to polygamy) remain unknown. During fertilization in the green alga Chlamydomonas, the plus gamete-specific membrane protein FUS1 is required for adhesion between the apically localized sites on the plasma membranes of plus and minus gametes that a...

  9. PrPC Undergoes Basal to Apical Transcytosis in Polarized Epithelial MDCK Cells.

    Directory of Open Access Journals (Sweden)

    Alexander Arkhipenko

    Full Text Available The Prion Protein (PrP is an ubiquitously expressed glycosylated membrane protein attached to the external leaflet of the plasma membrane via a glycosylphosphatidylinositol anchor (GPI. While the misfolded PrPSc scrapie isoform is the infectious agent of prion disease, the cellular isoform (PrPC is an enigmatic protein with unclear function. Of interest, PrP localization in polarized MDCK cells is controversial and its mechanism of trafficking is not clear. Here we investigated PrP traffic in MDCK cells polarized on filters and in three-dimensional MDCK cysts, a more physiological model of polarized epithelia. We found that, unlike other GPI-anchored proteins (GPI-APs, PrP undergoes basolateral-to-apical transcytosis in fully polarized MDCK cells. Following this event full-length PrP and its cleavage fragments are segregated in different domains of the plasma membrane in polarized cells in both 2D and 3D cultures.

  10. 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

    . 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......-week post-sampling, a power calculation revealed that over 900 patients are needed to show a difference of 9% between the two protocols tested. Conclusions. Future trials should be conducted using stringent protocols and as multi-centre trials for reaching the required information size....

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

    International Nuclear Information System (INIS)

    Geibel, M.A.; Schreiber, E.S.; Bracher, A.K.; Rasche, V.; Hell, E.; Ulrici, J.; Sailer, L.K.; Ozpeynirci, Y.

    2015-01-01

    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.

  12. In vitro comparison of apical microleakage following canal ...

    African Journals Online (AJOL)

    hope&shola

    2010-11-03

    Nov 3, 2010 ... The purpose of this study was to compare apical microleakage following canal obturation with lateral .... mm and the maximum was 6.69 mm. Statistical analysis of mean differences for independent groups (Independent samples t-test) showed that there was no ..... Clinical and radiographic evaluation of.

  13. Evaluation of the amount of apically extruded debris during ...

    African Journals Online (AJOL)

    2015-04-06

    Apr 6, 2015 ... (Dentsply Maillefer, Ballaigues, Switzerland). Root canal preparation and obturation. A size 15 K‑file (Dentsply Maillefer, Ballaigues, Switzerland) was placed into the canal until it was visible at the apical foramen. The working length (WL) was established as. 1 mm shorter than this length. The canals were ...

  14. 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.

  15. Shoot apical meristem arrest in brassica and tomato

    NARCIS (Netherlands)

    Jonge, de J.

    2013-01-01

    A pool of cells known as stem cells located in the center of the shoot apical meristem (SAM) are responsible to maintain meristematic activity throughout a plants life in order to produce organs. The maintenance of these stem cells is tightly controlled by a complex genetic and hormonal network. Any

  16. Histology of periapical lesions obtained during apical surgery.

    Science.gov (United States)

    Schulz, Malte; von Arx, Thomas; Altermatt, Hans Jörg; Bosshardt, Dieter

    2009-05-01

    The aim of this was to evaluate the histology of periapical lesions in teeth treated with periapical surgery. After root-end resection, the root tip was removed together with the periapical pathological tissue. Histologic sectioning was performed on calcified specimens embedded in methylmethacrylate (MMA) and on demineralized specimens embedded in LR White (Fluka, Buchs, Switzerland). The samples were evaluated with light and transmission electron microscopy (TEM). The histologic findings were classified into periapical abscesses, granulomas, or cystic lesions (true or pocket cysts). The final material comprised 70% granulomas, 23% cysts and 5% abscesses, 1% scar tissues, and 1% keratocysts. Six of 125 samples could not be used. The cystic lesions could not be subdivided into pocket or true cysts. All cysts had an epithelium-lined cavity, two of them with cilia-lined epithelium. These results show the high incidence of periapical granulomas among periapical lesions obtained during apical surgery. Periapical abscesses were a rare occasion. The histologic findings from samples obtained during apical surgery may differ from findings obtained by teeth extractions. A determination between pocket and true apical cysts is hardly possible when collecting samples by apical surgery.

  17. [Expression of interleukin-17F in human apical periodontitis lesions].

    Science.gov (United States)

    Sun, Hui; Li, Song

    2012-06-01

    To examine the expression of interleukin-17F (IL-17F) in apical periodontitis lesions. Human apical periodontitis lesions were collected after periapical surgery from November 1, 2009 to August 1, 2011. After histological processing, serial sectioning, HE staining, and microscopic analysis, 33 periapical cysts and 18 periapical granulomas were selected for immunohistochemical analysis for IL-17F. Five alveolar bone samples were obtained from a group of impacted third molars recommended for extraction. These bone samples did not show any inflammation and were used as normal samples. Differences in IL-17F expression between the tissues with periapical cyst and periapica granuloma were subsequently analyzed by rank-sum test. IL-17F was expressed in epithelial cells, lymphocytes, fibroblast and endothelial cells of both types of lesions. Both types of lesions presented increased IL-17F expression when compared to healthy periapical alveolar bone. The expression of IL-17F in apical cyst (0.2365 ± 0.0708) and apical granuloma (0.8852 ± 0.0576) was significantly different (Z = 3.735, P granuloma was significantly different.

  18. Phenotypic plasticity to altered apical bud temperature in Cucumis sativus

    NARCIS (Netherlands)

    Savvides, Andreas; Ieperen, van Wim; Dieleman, Janneke A.; Marcelis, Leo F.M.

    2017-01-01

    Many studies investigated temperature effects on leaf initiation and expansion by relating these processes to air temperature or the temperature of a specific organ (e.g. leaf temperature). In reality plant temperature is hardly ever equal to air temperature or spatially uniform. Apical bud

  19. Evaluation of the Amount of Apically Extruded Debris during ...

    African Journals Online (AJOL)

    Objective: To evaluate the amount of apically extruded debris during retreatment (with or without solvent) of root canals filled by two obturation techniques. Materials and Methods: Forty‑eight root canals were prepared using ProTaper Universal F3 and filled with Gutta‑percha and AH 26 sealer using single cone or lateral ...

  20. In vitro comparison of apical microleakage following canal ...

    African Journals Online (AJOL)

    hope&shola

    2010-11-03

    Nov 3, 2010 ... the cause of failing case in which the instructions were not followed and/or the goals of the technique were not achieved. Conclusion. Since the present study did not exhibit any differences in apical microleakage following canal obturation with lateral and thermoplasticized gutta-percha compaction techni-.

  1. [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.

  2. Apical dominance and growth in vitro of Alstroemeria

    NARCIS (Netherlands)

    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

  3. Adenylyl cyclase alpha and cAMP signaling mediate Plasmodium sporozoite apical regulated exocytosis and hepatocyte infection.

    Directory of Open Access Journals (Sweden)

    Takeshi Ono

    2008-02-01

    Full Text Available Malaria starts with the infection of the liver of the host by Plasmodium sporozoites, the parasite form transmitted by infected mosquitoes. Sporozoites migrate through several hepatocytes by breaching their plasma membranes before finally infecting one with the formation of an internalization vacuole. Migration through host cells induces apical regulated exocytosis in sporozoites. Here we show that apical regulated exocytosis is induced by increases in cAMP in sporozoites of rodent (P. yoelii and P. berghei and human (P. falciparum Plasmodium species. We have generated P. berghei parasites deficient in adenylyl cyclase alpha (ACalpha, a gene containing regions with high homology to adenylyl cyclases. PbACalpha-deficient sporozoites do not exocytose in response to migration through host cells and present more than 50% impaired hepatocyte infectivity in vivo. These effects are specific to ACalpha, as re-introduction of ACalpha in deficient parasites resulted in complete recovery of exocytosis and infection. Our findings indicate that ACalpha and increases in cAMP levels are required for sporozoite apical regulated exocytosis, which is involved in sporozoite infection of hepatocytes.

  4. 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.

  5. CLINICAL EFFECTIVENESS OF TREATMENT THE PATIENTS WITH CHRONIC APICAL PERIODONTITIS

    Directory of Open Access Journals (Sweden)

    N. G. Gadzhula

    2017-02-01

    Full Text Available Background. The success of endodontic treatment is provided by a thorough instrumental and antiseptic treatment of infected root canals, and it depends on the composition of filling material, the degree of adhesion to dentin, hermetic obturation of apical foramen, solubility of sealer. Objective. The study was aimed to evaluate the effectiveness of root canal obturation with BioRootTM RCS sealer in the treatment of patients with chronic apical periodontitis. Methods. Endodontic treatment of 23 teeth in 20 patients with chronic apical periodontitis by method of lateral compaction of gutta-percha was carried out. In the main group root canals were obturated with BioRootTM RCS, in the control group the canals were filled with Apexit Plus. The percentage of efficient or non-efficient cases was evaluated on the basis of radiographic comparison of treated chronic apical periodontitis immediately after obturation, in three, six months and one year. Radiographic conditions were defined as existing state, improvement and worsening. Results. In a year of dynamic evaluation the final results were: in the main group – 54.55% of the patients had complete bone healing, in 27.27% of cases the focus of bone destruction was decreased by ½ or more of the initial sizes, 18.18% – resorption lesion was decreased by less than ½; in the control group – 33.33% of improvement, 25.0% of existing state and 41.66% of worsening. Conclusions. BioRootTM RCS using for root canals obturation in the treatment of chronic apical periodontitis we proved the high effectiveness of the treatment undertaken: complete healing or improvement of radiographic conditions of periapical bone destruction with X-ray signs of bone regeneration.

  6. AntigenMap 3D: an online antigenic cartography resource.

    Science.gov (United States)

    Barnett, J Lamar; Yang, Jialiang; Cai, Zhipeng; Zhang, Tong; Wan, Xiu-Feng

    2012-05-01

    Antigenic cartography is a useful technique to visualize and minimize errors in immunological data by projecting antigens to 2D or 3D cartography. However, a 2D cartography may not be sufficient to capture the antigenic relationship from high-dimensional immunological data. AntigenMap 3D presents an online, interactive, and robust 3D antigenic cartography construction and visualization resource. AntigenMap 3D can be applied to identify antigenic variants and vaccine strain candidates for pathogens with rapid antigenic variations, such as influenza A virus. http://sysbio.cvm.msstate.edu/AntigenMap3D

  7. Antigens of Streptococcus sanguis

    Science.gov (United States)

    Rosan, Burton

    1973-01-01

    An antigenic analysis of the alpha-hemolytic streptococci isolated from dental plaque was performed by use of antisera against a strain of Streptococcus sanguis (M-5) which was isolated from dental plaque. Immunoelectrophoretic and Ouchterlony tests of Rantz and Randall extracts of 45 strains gave positive reactions with the M-5 antisera. These strains represented 60% of the strains tested. The number of antigens which could be identified in these extracts varied from one to five and were designated a to e. The a antigen was found in 36 of the strains tested, including reference strains of S. sanguis and the group H streptococci. The strains reacting with the M-5 antisera were divided into two majors types: type I consisted of 23 strains in which the a antigen was found alone or with one or more of the c, d, and e antigens; type II consisted of 13 strains in which both the a and b antigens were found with or without one or more of the c, d, and e antigens. The remaining strains contained, either singly or in combination, the b, c, d, and e antigens but not the a antigen. Biochemical tests of representatives of each serotype and reference strains indicated that strains reacting with M-5 antisera were S. sanguis. These findings suggest that S. sanguis strains share common physiological and serological properties. Images PMID:4633291

  8. An Atypical Case of Apical Hypertrophic Cardiomyopathy: Absence of Giant T Waves in spite of Extreme Apical Wall Hypertrophy

    Directory of Open Access Journals (Sweden)

    Elias Sanidas

    2015-01-01

    Full Text Available Apical hypertrophic cardiomyopathy is an uncommon variant of hypertrophic cardiomyopathy, with hypertrophy mainly affecting the apex of the left ventricle. We hereby describe a case of an octogenarian female patient who was randomly diagnosed with AHCM due to other comorbidities.

  9. [Bacterial localization in apical cementum at the epithelial insertion using scanning electron microscopy].

    Science.gov (United States)

    García Núñez, J A; Herrera, I; Cerezo Lapiedra, R; Santa María, I

    1989-02-01

    Extracted teeth due to consequence of chronic periodontitis of adult are fractured and the apical cementum to junction epithelium is examined under S.E.M. (scanning electron microscopy) being found bacterias forms inside niches of the apical cementum.

  10. The MRP4/ABCC4 gene encodes a novel apical organic anion transporter in human kidney proximal tubules: putative efflux pump for urinary cAMP and cGMP.

    NARCIS (Netherlands)

    Aubel, R.A.M.H. van; Smeets, P.H.E.; Peters, J.G.P.; Bindels, R.J.M.; Russel, F.G.M.

    2002-01-01

    The cyclic nucleotides cAMP and cGMP play key roles in cellular signaling and the extracellular regulation of fluid balance. In the kidney, cAMP is excreted across the apical proximal tubular membrane into urine, where it reduces phosphate reabsorption through a dipyridamole-sensitive mechanism that

  11. Apical organs in echinoderm larvae: insights into larval evolution in the Ambulacraria.

    Science.gov (United States)

    Byrne, Maria; Nakajima, Yoko; Chee, Francis C; Burke, Robert D

    2007-01-01

    The anatomy and cellular organization of serotonergic neurons in the echinoderm apical organ exhibits class-specific features in dipleurula-type (auricularia, bipinnaria) and pluteus-type (ophiopluteus, echinopluteus) larvae. The apical organ forms in association with anterior ciliary structures. Apical organs in dipleurula-type larvae are more similar to each other than to those in either of the pluteus forms. In asteroid bipinnaria and holothuroid auricularia the apical organ spans ciliary band sectors that traverse the anterior-most end of the larvae. The asteroid apical organ also has prominent bilateral ganglia that connect with an apical network of neurites. The simple apical organ of the auricularia is similar to that in the hemichordate tornaria larva. Apical organs in pluteus forms differ markedly. The echinopluteus apical organ is a single structure on the oral hood between the larval arms comprised of two groups of cells joined by a commissure and its cell bodies do not reside in the ciliary band. Ophioplutei have a pair of lateral ganglia associated with the ciliary band of larval arms that may be the ophiuroid apical organ. Comparative anatomy of the serotonergic nervous systems in the dipleurula-type larvae of the Ambulacraria (Echinodermata+Hemichordata) suggests that the apical organ of this deuterostome clade originated as a simple bilaterally symmetric nerve plexus spanning ciliary band sectors at the anterior end of the larva. From this structure, the apical organ has been independently modified in association with the evolution of class-specific larval forms.

  12. 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...

  13. Eosinofil Sel Penyaji Antigen

    Directory of Open Access Journals (Sweden)

    Safari Wahyu Jatmiko

    2015-04-01

    Full Text Available Sel eosinofil merupakan jenis sel lekosit yang terlibat dalam berbagai patogenesis penyakit. Sel eosinofil pada awalnya dikenal sebagai sel efektor  dari sistem imunitas alamiah. Akan tetapi, kemampuan sel eosinofil dalam memfagositosis patogen menimbulkan dugaan bahwa sel eosinofil ikut berperan sebagai sel penyaji antigen. Hal ini dianalogikan dengan sel makrofag dan sel dendritik yang bisa memfagositosis dan menyajikan antigen sebagai hasil dari degradasi patogen yang difagositosis. Untuk menjawab permasalahan ini, penulis melakukan penelusuran artikel tentang eosinofil sebagai sel penyaji antigen melalui US National Library of Medicine National Institute of Healthdengan kata kunci eoshinophil dan antigen presenting cell. Hasil penelusuran adalah ditemukannya 10 artikel yang relevan dengan topik. Hasil dari sintesis kesepuluh jurnal tersebut adalah sel eosinofil mampu berperan sebagai sel penyaji antigen yang profesional (professionalantigenpresentng cell

  14. [Enterobacterial antigen in human peripheral blood lymphocytes].

    Science.gov (United States)

    Faure-Fontenla, M A; García-Tamayo, F

    1989-11-01

    The following study has as prior history the research reports which have shown the existence of an antigenic tissue deposit in gram-negative enterobacteria. The antigens of the enterobacteria have also been found in the lymphocytic membranes and cytoplasm. Since intestinal lymphoid tissue cells can recirculate by means of the thoracic duct to the peripheral venous system, it was proposed that the circulating lymphocytes in healthy people could also contain small amounts of a common enterobacterial antigen. The study was carried out in 15 human venous blood samples, of which the lymphocytic population was separated to later be used in the preparation of 15 alcohol soluble extracts. This material was used for inhibiting the immuno-hemolysis assay in three occasions in order to show the presence of antigens shared by different enterobacterias, using as reference a fraction separated from the LPS of Escherichia coli 08. The results showed that the human lymphocytes also had antigenic determinants common to gram-negative bacteria.

  15. Renoguanylin stimulates apical CFTR translocation and decreases HCO3-secretion through PKA activity in the Gulf toadfish (Opsanus beta).

    Science.gov (United States)

    Ruhr, Ilan M; Schauer, Kevin L; Takei, Yoshio; Grosell, Martin

    2018-03-26

    The guanylin peptides - guanylin, uroguanylin and renoguanylin (RGN) - are endogenously produced hormones in teleost fish enterocytes that are activators of guanylyl cyclase-C (GC-C) and are potent modulators of intestinal physiology, particularly in seawater teleosts. Most notably, they reverse normal net ion-absorbing mechanisms that are vital to water absorption, an important process for seawater teleost survival. The role of guanylin-peptide stimulation of the intestine remains unclear, but it is hypothesized to facilitate the removal of solids from the intestine by providing fluid to enable their removal by peristalsis. The present study used one member of this group of peptides - RGN - to provide evidence for the prominent role that protein kinase A (PKA) plays in mediating the effects of guanylin-peptide stimulation in the posterior intestine of the Gulf toadfish ( Opsanus beta ). Protein kinase G was found to not mediate the intracellular effects of RGN, despite previous evidence showing that GC-C activation leads to higher cyclic guanosine monophosphate formation. RGN reversed the absorptive short-circuit current and increased conductance in the Gulf toadfish intestine. These effects are correlated to increased trafficking of the cystic fibrosis transmembrane conductance regulator (CFTR) Cl - channel to the apical membrane, which is negated by PKA inhibition. Moreover, RGN decreased HCO 3 - secretion, likely by limiting apical HCO 3 - /Cl - exchange (possibly by reducing SLC26a6 activity), a reduction that was enhanced by PKA inhibition. RGN seems to alter PKA activity in the posterior intestine to recruit CFTR to the apical membrane and reduce HCO 3 - secretion. © 2018. Published by The Company of Biologists Ltd.

  16. The Effect of Canal Contamination with Saliva on Apical Sealing

    Directory of Open Access Journals (Sweden)

    S Sabaghi

    2014-08-01

    Methods: In this laboratory study, 58 human uni-root teeth were cleaned and shaped for obturation with gutta percha and sealer AH26. In the case group, specimens were contaminated with human saliva immediately before obturation, whereas the teeth in the control group were kept dry. All canals were filled by lateral condensation technique. Moreover, the teeth were placed in methylene blue dye for 3 days. Dye penetration was measured using a stereomicrosope. As a matter of fact, the study data were analyzed via utilizing t-test. Results: A significant difference was found between the two groups in regard with the apical leakage(P<0.001. The microleakage mean of dye in the dry group was 3/48mm, whereas it was 6/36mm in the saliva contaminated group. Conclusion: The study findings revealed that complete drying of canal can improve apical sealing.

  17. 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.

  18. Characterization of human apical papilla-derived stem cells.

    Science.gov (United States)

    Cantore, S; Ballini, A; De Vito, D; Martelli, F S; Georgakopoulos, I; Almasri, M; Dibello, V; Altini, V; Farronato, G; Dipalma, G; Farronato, D; Inchingolo, F

    2017-01-01

    Dental tissues represent an alternative and promising source of post-natal Mesenchymal stem cells (MSCs) for tissue engineering. Furthermore, dental stem cells from apical papilla (SCAPs) cells can be obtained from the wisdom tooth which is unnecessary for human masticatory function and frequently extracted for orthodontic reasons or dysodontiasis. More precisely, apical papilla is the immature, mostly uncalcified, precursor of the tooth root, therefore is composed of more undifferentiated cells than dental pulp. In addition, tooth extraction, especially by piezosurgery technique, can be considered less invasive in comparison to bone marrow or other tissues biopsy. Our work is aimed to investigate the safety of and predictable procedure on surgical immature third molar extraction and to provide new insight on SCAP research for future biomedical applications. The isolated cells were examined for stem cell properties by analyzing their colony-forming efficiency, differentiation characteristics and the expression of MSC markers.

  19. 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. Copyright © 2014 Sociedade Portuguesa de Cardiologia. Published by Elsevier España. All rights reserved.

  20. Bidirectional apical-basal traffic of the cation-independent mannose-6-phosphate receptor in brain endothelial cells.

    Science.gov (United States)

    Siupka, Piotr; Hersom, Maria Ns; Lykke-Hartmann, Karin; Johnsen, Kasper B; Thomsen, Louiza B; Andresen, Thomas L; Moos, Torben; Abbott, N Joan; Brodin, Birger; Nielsen, Morten S

    2017-07-01

    Brain capillary endothelium mediates the exchange of nutrients between blood and brain parenchyma. This barrier function of the brain capillaries also limits passage of pharmaceuticals from blood to brain, which hinders treatment of several neurological disorders. Receptor-mediated transport has been suggested as a potential pharmaceutical delivery route across the brain endothelium, e.g. reports have shown that the transferrin receptor (TfR) facilitates transcytosis of TfR antibodies, but it is not known whether this recycling receptor itself traffics from apical to basal membrane in the process. Here, we elucidate the endosomal trafficking of the retrograde transported cation-independent mannose-6-phosphate receptor (MPR300) in primary cultures of brain endothelial cells (BECs) of porcine and bovine origin. Receptor expression and localisation of MPR300 in the endo-lysosomal system and trafficking of internalised receptor are analysed. We also demonstrate that MPR300 can undergo bidirectional apical-basal trafficking in primary BECs in co-culture with astrocytes. This is, to our knowledge, the first detailed study of retrograde transported receptor trafficking in BECs, and the study demonstrates that MPR300 can be transported from the luminal to abluminal membrane and reverse. Such trafficking of MPR300 suggests that retrograde transported receptors in general may provide a mechanism for transport of pharmaceuticals into the brain.

  1. Helicobacter pylori dwelling on the apical surface of gastrointestinal epithelium damages the mucosal barrier through direct contact.

    Science.gov (United States)

    Zhang, Chen; Zhang, Hongyu; Yu, Lu; Cao, Yi

    2014-10-01

    Epithelial junctions and mucins compose a major portion of the mucosal barrier. Helicobacter pylori (H. pylori) infections induce alterations of the tight junctions and adherens junctions in epithelial cells, although the precise mechanisms underlying this process are not fully understood. The expression of adhesion molecules and MUC1 was systematically investigated in gastrointestinal epithelial cells infected with H. pylori in vitro and in vivo. Furthermore, we developed several new in vitro methods to study the relationships between the bacterium and the dysfunction of tight junctions using Boyden Chambers. The expression of a series of junctional molecules and MUC1 decreased in the cultured cells that were infected with H. pylori. According to the degree of damage at the tight junctions, direct contact of H. pylori with the apical membrane of the cells resulted in the greatest increase in permeability compared to basal membrane binding or non-binding of H. pylori to the cells. Similarly, we noted that H. pylori infection could reduce the expression and glycosylation of MUC1. Helicobacter pylori dwelling on the apical surface of the gastrointestinal epithelium could directly induce serious injury of the mucosal barrier, and the new methods outlined here, based on the Boyden Chamber system, could be very useful for studying the relationships between bacteria and their target cells. © 2014 John Wiley & Sons Ltd.

  2. Apically Extruded Sealers: Fate and Influence on Treatment Outcome.

    Science.gov (United States)

    Ricucci, Domenico; Rôças, Isabela N; Alves, Flávio R F; Loghin, Simona; Siqueira, José F

    2016-02-01

    This retrospective study evaluated cases of unintentional overfillings for the fate of the extruded sealers and their influence on treatment outcome. One hundred five teeth treated by a single operator and exhibiting overfillings in the postobturation radiograph were included in the study. Seventy-five teeth exhibited apical periodontitis lesions at the time of treatment. Sealers included Pulp Canal Sealer (Sybron Dental, Orange, CA), PCS Extended Working Time-EWT (Sybron Dental), Tubli-Seal (Sybron Endo), Endomethasone (Septodont, Saint-Maur-des-Fossés, France), AH Plus (DeTrey GmbH, Konstanz, Germany), and Apexit (Ivoclar Vivadent, Schaan, Lichtenstein). Recall radiographs were compared with immediate postobturation films for removal of the extruded material and status of the periradicular tissues. Data were grouped as 1-, 2- and >4-year recall and statistically analyzed using the chi-square and Fisher exact tests. As for the sealers' fate, the only statistically significant differences at the 1-year recall were observed when comparing Tubli-Seal with AH Plus, Apexit, and Endomethasone (P .05). Data from the >4-year recall revealed that 79% of the teeth with apical periodontitis lesions at the time of treatment had healed in comparison with 100% of the teeth with no apical periodontitis (P extruded sealers were predictably removed from the periradicular tissues. Treatment outcome was not significantly affected by the type of extruded sealer. A significantly better outcome was observed for teeth with no lesion in comparison with teeth with apical periodontitis. Copyright © 2016 American Association of Endodontists. Published by Elsevier Inc. All rights reserved.

  3. Impact of osmotic dehydration on the encapsulated apices survival ...

    African Journals Online (AJOL)

    L-1 activated charcoal) prior to encapsulation in alginate (3%) beads with calcium chloride (1.32M). Then, the apices were exposed to osmotic dehydration with two concentrations of sucrose (0.75M and 1.25M) at two durations (24h and 40h) before their culture in M2 medium (MS + 2mg.L-1 BAP, 100?g.L-1 d'ANA and 2g.

  4. Bacterial membrane proteomics.

    Science.gov (United States)

    Poetsch, Ansgar; Wolters, Dirk

    2008-10-01

    About one quarter to one third of all bacterial genes encode proteins of the inner or outer bacterial membrane. These proteins perform essential physiological functions, such as the import or export of metabolites, the homeostasis of metal ions, the extrusion of toxic substances or antibiotics, and the generation or conversion of energy. The last years have witnessed completion of a plethora of whole-genome sequences of bacteria important for biotechnology or medicine, which is the foundation for proteome and other functional genome analyses. In this review, we discuss the challenges in membrane proteome analysis, starting from sample preparation and leading to MS-data analysis and quantification. The current state of available proteomics technologies as well as their advantages and disadvantages will be described with a focus on shotgun proteomics. Then, we will briefly introduce the most abundant proteins and protein families present in bacterial membranes before bacterial membrane proteomics studies of the last years will be presented. It will be shown how these works enlarged our knowledge about the physiological adaptations that take place in bacteria during fine chemical production, bioremediation, protein overexpression, and during infections. Furthermore, several examples from literature demonstrate the suitability of membrane proteomics for the identification of antigens and different pathogenic strains, as well as the elucidation of membrane protein structure and function.

  5. The Effect of Canal Contamination with Saliva on Apical Sealing

    Directory of Open Access Journals (Sweden)

    S Sabaghi

    2014-08-01

    Full Text Available Introduction: Root canal obturation aims at sealing the root canal system to prevent re-contamination of canal and periapical space. Presence of moisture in canal before obturation may posit a negative effect on the quality of canal sealing. Therefore, this study was conducted to investigate the effect of canal contamination with saliva on apical microleakage. Methods: In this laboratory study, 58 human uni-root teeth were cleaned and shaped for obturation with gutta percha and sealer AH26. In the case group, specimens were contaminated with human saliva immediately before obturation, whereas the teeth in the control group were kept dry. All canals were filled by lateral condensation technique. Moreover, the teeth were placed in methylene blue dye for 3 days. Dye penetration was measured using a stereomicrosope. As a matter of fact, the study data were analyzed via utilizing t-test. Results: A significant difference was found between the two groups in regard with the apical leakage(P<0.001. The microleakage mean of dye in the dry group was 3/48mm, whereas it was 6/36mm in the saliva contaminated group. Conclusion: The study findings revealed that complete drying of canal can improve apical sealing.

  6. 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

    retained in the endoplasmic reticulum or partially secreted into the medium. Coexpression with AMN led to efficient transport to the apical cell surface of the cubilin constructs, which included the EGF domains, and prevented release into the medium. AMN co-precipitated with cubilin and co-localized......Cubilin is a peripheral apical membrane receptor for multiple ligands that are taken up in several absorptive epithelia. Recently, amnionless (AMN) was identified to form a functional receptor complex with cubilin. By expression in transfected polarized MDCK cells of AMN and several cubilin...... 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...

  7. Endoplasmic reticulum resident protein of 90 kilodaltons associates with the T- and B-cell antigen receptors and major histocompatibility complex antigens during their assembly

    NARCIS (Netherlands)

    Hochstenbach, F.; DAVID, V.; WATKINS, S.; Brenner, M. B.

    1992-01-01

    In the endoplasmic reticulum (ER), newly synthesized subunits of the T-cell antigen receptor (TCR), membrane-bound immunoglobulin (mIg), and major histocompatibility complex (MHC) class I antigens must fold correctly and assemble completely into multimeric protein complexes prior to transport to the

  8. Carcinoembryonic antigen (CEA)

    International Nuclear Information System (INIS)

    Ephraim, K.H.; Cox, P.H.; Hamer, C.J.A. v.d.; Berends, W.; Delhez, H.

    1977-01-01

    The carcinoembryonic antigen (CEA) is a complex of antigen determinants and also the carrier of these determinants. Chemically it is a glycoprotein. Its occurrence in blood serum or urine is correlated with malignant disease. Several radioimmunoassays (RIA) have been developed, one by Hoffmann-Laroche and one by the Rotterdam Radiotherapeutic Institute. Both methods and the Hoffmann assay kit are tested. Specifications are given for isolation of the antigen, preparation of the antiserum, and the execution of the RIA. Biochemical and clinical aspects are discussed

  9. 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....

  10. Biobased Membrane

    NARCIS (Netherlands)

    Koenders, E.A.B.; Zlopasa, J.; Picken, S.J.

    2015-01-01

    The present invention is in the field of a composition for forming a bio-compatible membrane applicable to building material, such as concrete, cement, etc., to a meth od of applying said composition for forming a bio-compatible membrane, a biocompatible membrane, use of said membrane for various

  11. Presentation of phagocytosed antigens by MHC class I and II

    Science.gov (United States)

    Mantegazza, Adriana R.; Magalhaes, Joao G.; Amigorena, Sebastian; Marks, Michael S.

    2012-01-01

    Phagocytosis provides innate immune cells with a mechanism to take up and destroy pathogenic bacteria, apoptotic cells and other large particles. In some cases, however, peptide antigens from these particles are preserved for presentation in association with major histocompatibility complex (MHC) class I or class II molecules in order to stimulate antigen-specific T cells. Processing and presentation of antigens from phagosomes presents a number of distinct challenges relative to antigens internalized by other means; While bacterial antigens were among the first discovered to be presented to T cells, analyses of the cellular mechanisms by which peptides from phagocytosed antigens assemble with MHC molecules and by which these complexes are then expressed at the plasma membrane have lagged behind those of conventional model soluble antigens. In this review, we cover recent advances in our understanding of these processes, including the unique cross-presentation of phagocytosed antigens by MHC class I molecules, and in their control by signaling modalities in phagocytic cells. PMID:23127154

  12. Does apical root resection in endodontic microsurgery jeopardize the prosthodontic prognosis?

    Directory of Open Access Journals (Sweden)

    Sin-Yeon Cho

    2013-05-01

    Full Text Available Apical surgery cuts off the apical root and the crown-to-root ratio becomes unfavorable. Crown-to-root ratio has been applied to periodontally compromised teeth. Apical root resection is a different matter from periodontal bone loss. The purpose of this paper is to review the validity of crown-to-root ratio in the apically resected teeth. Most roots have conical shape and the root surface area of coronal part is wider than apical part of the same length. Therefore loss of alveolar bone support from apical resection is much less than its linear length.The maximum stress from mastication concentrates on the cervical area and the minimum stress was found on the apical 1/3 area. Therefore apical root resection is not so harmful as periodontal bone loss. Osteotomy for apical resection reduces longitudinal width of the buccal bone and increases the risk of endo-perio communication which leads to failure. Endodontic microsurgery is able to realize 0 degree or shallow bevel and precise length of root resection, and minimize the longitudinal width of osteotomy. The crown-to-root ratio is not valid in evaluating the prosthodontic prognosis of the apically resected teeth. Accurate execution of endodontic microsurgery to preserve the buccal bone is essential to avoid endo-perio communication.

  13. Does apical root resection in endodontic microsurgery jeopardize the prosthodontic prognosis?

    Science.gov (United States)

    Cho, Sin-Yeon; Kim, Euiseong

    2013-05-01

    Apical surgery cuts off the apical root and the crown-to-root ratio becomes unfavorable. Crown-to-root ratio has been applied to periodontally compromised teeth. Apical root resection is a different matter from periodontal bone loss. The purpose of this paper is to review the validity of crown-to-root ratio in the apically resected teeth. Most roots have conical shape and the root surface area of coronal part is wider than apical part of the same length. Therefore loss of alveolar bone support from apical resection is much less than its linear length.The maximum stress from mastication concentrates on the cervical area and the minimum stress was found on the apical 1/3 area. Therefore apical root resection is not so harmful as periodontal bone loss. Osteotomy for apical resection reduces longitudinal width of the buccal bone and increases the risk of endo-perio communication which leads to failure. Endodontic microsurgery is able to realize 0 degree or shallow bevel and precise length of root resection, and minimize the longitudinal width of osteotomy. The crown-to-root ratio is not valid in evaluating the prosthodontic prognosis of the apically resected teeth. Accurate execution of endodontic microsurgery to preserve the buccal bone is essential to avoid endo-perio communication.

  14. Bacterial communities associated with apical periodontitis and dental implant failure

    Science.gov (United States)

    Dingsdag, Simon; Nelson, Stephen; Coleman, Nicholas V.

    2016-01-01

    Background Previously, we demonstrated that bacteria reside in apparently healed alveolar bone, using culture and Sanger sequencing techniques. Bacteria in apparently healed alveolar bone may have a role in peri-implantitis and dental implant failure. Objective To compare bacterial communities associated with apical periodontitis, those colonising a failed implant and alveolar bone with reference biofilm samples from healthy teeth. Methods and results The study consisted of 196 samples collected from 40 patients undergoing routine dental implant insertion or rehabilitation. The bacterial 16S ribosomal DNA sequences were amplified. Samples yielding sufficient polymerase chain reaction product for further molecular analyses were subjected to terminal restriction fragment length polymorphism (T-RFLP; 31 samples) and next generation DNA sequencing (454 GS FLX Titanium; 8 samples). T-RFLP analysis revealed that the bacterial communities in diseased tissues were more similar to each other (p<0.049) than those from the healthy reference samples. Next generation sequencing detected 13 bacterial phyla and 373 putative bacterial species, revealing an increased abundance of Gram-negative [Prevotella, Fusobacterium (p<0.004), Treponema, Veillonellaceae, TG5 (Synergistetes)] bacteria and a decreased abundance of Gram-positive [(Actinomyces, Corynebacterium (p<0.008)] bacteria in the diseased tissue samples (n=5) relative to reference supragingival healthy samples (n=3). Conclusion Increased abundances of Prevotella, Fusobacterium and TG5 (Synergistetes) were associated with apical periodontitis and a failed implant. A larger sample set is needed to confirm these trends and to better define the processes of bacterial pathogenesis in implant failure and apical periodontitis. The application of combined culture-based, microscopic and molecular technique-based approaches is suggested for future studies. PMID:27834171

  15. Progressive, generalized, apical idiopathic root resorption and hypercementosis.

    Science.gov (United States)

    Schätzle, Marc; Tanner, Sandro D; Bosshardt, Dieter D

    2005-11-01

    Root resorption is a multifactorial process that results in loss of tooth structure. The causes of root resorption may vary, leading to several types of resorptions. Some factors have been identified and may be categorized into physiological resorption, local factors, systemic conditions, and idiopathic resorptions. The objective of this report was to present a case of a 17-year-old white female with progressive, generalized, apical idiopathic root resorption followed up for 34 months. Two panoramic radiographs, 14 and 34 months after initial clinical and radiological examinations, showed the rapid progression of apical root resorption. Two molars, teeth #15 and #16, which had to be extracted, and a bone sample from the distal aspect of tooth #15 were processed for histologic analysis. Two millimeters apical to the cemento-enamel junction, an abrupt increase in the cementum thickness was noted, amounting to 300 and 800 microm in teeth #15 and #16, respectively. The thickening of the cementum layer was due to an accelerated deposition of cellular intrinsic fiber cementum. An unusually high number of mineralization foci were observed in association with acellular extrinsic fiber cementum, and both free and fused cementicles were seen. In contrast to tooth #16, tooth #15 revealed extensive dentin replacement by a bone-like and a cementum-like tissue. Furthermore, ankylosis was demonstrated in tooth #15 and confirmed in the bone sample. At present, there is no preventive or therapeutic regimen for the type of root resorption seen in this case report. Treatment usually consists of the extraction of teeth with advanced lesions.

  16. 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.

  17. Immunoexpression of interleukin 17 in apical periodontitis lesions.

    Science.gov (United States)

    Ajuz, Natasha C; Antunes, Henrique; Mendonça, Thais A; Pires, Fábio R; Siqueira, José F; Armada, Luciana

    2014-09-01

    Interleukin (IL)-17 expression has been detected in apical periodontitis lesions, but its role in the disease process remains unclear. The present study compared the expression of IL-17 in periradicular cysts and granulomas and evaluated the association of this cytokine with clinical and radiographic findings. Apical periodontitis lesions (18 cysts and 20 granulomas) were obtained from 38 patients subjected to periradicular surgery. Some clinical, radiographic, and cone-beam computed tomographic features were recorded. Silanized slides containing paraffin sections were used for the immunohistochemical reactions using anti-IL-17 antibody. Image analysis was performed using an optical microscope, and each sample was divided into 5 high-power fields, which were evaluated for the expression of IL-17 in the epithelium and connective tissues. Results were evaluated for correlations with the lesion size and the occurrence of symptoms and sinus tract. Expression of IL-17 was significantly higher in cysts than in granulomas (P = .02). Among the periradicular cysts, a thin epithelium showed significantly increased labeling for IL-17 when compared with a hyperplastic epithelium (P = .003). IL-17 expression was usually associated with focal accumulations of polymorphonuclear leukocytes. No association of IL-17 expression with symptoms, sinus tract, or lesion size was observed (P > .05). The present study reinforces the notion that IL-17 may take part in the pathogenesis of apical periodontitis lesions. A role in the exacerbation of chronic inflammation and cyst formation is suspected. Further studies are required to shed light on the specific functions of IL-17 in periradicular inflammatory processes. Copyright © 2014 American Association of Endodontists. Published by Elsevier Inc. All rights reserved.

  18. Bacterial communities associated with apical periodontitis and dental implant failure

    Directory of Open Access Journals (Sweden)

    Simon Dingsdag

    2016-11-01

    Full Text Available Background: Previously, we demonstrated that bacteria reside in apparently healed alveolar bone, using culture and Sanger sequencing techniques. Bacteria in apparently healed alveolar bone may have a role in peri-implantitis and dental implant failure. Objective: To compare bacterial communities associated with apical periodontitis, those colonising a failed implant and alveolar bone with reference biofilm samples from healthy teeth. Methods and results: The study consisted of 196 samples collected from 40 patients undergoing routine dental implant insertion or rehabilitation. The bacterial 16S ribosomal DNA sequences were amplified. Samples yielding sufficient polymerase chain reaction product for further molecular analyses were subjected to terminal restriction fragment length polymorphism (T-RFLP; 31 samples and next generation DNA sequencing (454 GS FLX Titanium; 8 samples. T-RFLP analysis revealed that the bacterial communities in diseased tissues were more similar to each other (p<0.049 than those from the healthy reference samples. Next generation sequencing detected 13 bacterial phyla and 373 putative bacterial species, revealing an increased abundance of Gram-negative [Prevotella, Fusobacterium (p<0.004, Treponema, Veillonellaceae, TG5 (Synergistetes] bacteria and a decreased abundance of Gram-positive [(Actinomyces, Corynebacterium (p<0.008] bacteria in the diseased tissue samples (n=5 relative to reference supragingival healthy samples (n=3. Conclusion: Increased abundances of Prevotella, Fusobacterium and TG5 (Synergistetes were associated with apical periodontitis and a failed implant. A larger sample set is needed to confirm these trends and to better define the processes of bacterial pathogenesis in implant failure and apical periodontitis. The application of combined culture-based, microscopic and molecular technique-based approaches is suggested for future studies.

  19. Locus of the apices of projectile trajectories under constant drag

    Science.gov (United States)

    Hernández-Saldaña, H.

    2017-11-01

    Using the hodograph method, we present an analytical solution for projectile coplanar motion under constant drag, parametrised by the velocity angle. We find the locus formed by the apices of the projectile trajectories, and discuss its implementation for the motion of a particle on an inclined plane in presence of Coulomb friction. The range and time of flight are obtained numerically, and we find that the optimal launching angle is smaller than in the drag-free case. This is a good example of a problem with constant dissipation of energy that includes curvature; it is appropriate for intermediate courses of mechanics.

  20. Pyogenic granuloma subsequent to apical fenestration of a primary tooth.

    Science.gov (United States)

    Aguiló, Luz; Bagán, José V

    2002-05-01

    The authors present two case reports of patients exhibiting pyogenic granulomas in the maxillary labial mucosa, which were related to an apical fenestration of a primary incisor. Several researchers have reported that the gingival wound and surrounding inflammatory tissue typically heal spontaneously after extraction of a fenestrated primary tooth. However, in the cases presented here, the gingival lesion did not heal after the fenestrated teeth were extracted. After extracting fenestrated teeth, clinicians need to examine the labial area at a follow-up appointment to ensure that the gingival hyperplasia heals properly. The authors suggest performing curettage of the surrounding abnormal tissue at the time of the tooth extraction.

  1. Antigenic Variation of TprK Facilitates Development of Secondary Syphilis

    OpenAIRE

    Reid, Tara B.; Molini, Barbara J.; Fernandez, Mark C.; Lukehart, Sheila A.

    2014-01-01

    Although primary syphilis lesions heal spontaneously, the infection is chronic, with subsequent clinical stages. Healing of the primary chancre occurs as antibodies against outer membrane antigens facilitate opsonophagocytosis of the bacteria by activated macrophages. TprK is an outer membrane protein that undergoes antigenic variation at 7 variable regions, and variants are selected by immune pressure. We hypothesized that individual TprK variants escape immune clearance and seed new dissemi...

  2. Foetal antigen 2 (FA2) in the stromal reaction induced by breast carcinoma

    DEFF Research Database (Denmark)

    Rasmussen, H B; Teisner, B; Andersen, J A

    1992-01-01

    An indirect immunoperoxidase technique was used to examine the distribution of foetal antigen 2 (FA2), a recently described basement membrane (BM)-associated antigen, in invasive breast carcinoma (n = 34), fibroadenoma (n = 5) and normal breast tissue (n = 5), and to compare its distribution...

  3. 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...

  4. Saposins utilize two strategies for lipid transfer and CD1 antigen presentation

    NARCIS (Netherlands)

    Leon, Luis; Tatituri, Raju V. V.; Grenha, Rosa; Sun, Ying; Barral, Duarte C.; Minnaard, Adriaan J.; Bhowruth, Veemal; Veerapen, Natacha; Besra, Gurdyal S.; Kasmar, Anne; Peng, Wei; Moody, D. Branch; Grabowski, Gregory A.; Brenner, Michael B.

    2012-01-01

    Transferring lipid antigens from membranes into CD1 antigen-presenting proteins represents a major molecular hurdle necessary for T-cell recognition. Saposins facilitate this process, but the mechanisms used are not well understood. We found that saposin B forms soluble saposin protein-lipid

  5. Molecular typing of human platelet and neutrophil antigens (HPA and HNA)

    NARCIS (Netherlands)

    Veldhuisen, Barbera; Porcelijn, Leendert; Ellen van der Schoot, C.; de Haas, Masja

    2014-01-01

    Genotyping is an important tool in the diagnosis of disorders involving allo-immunisation to antigens present on the membranes of platelets and neutrophils. To date 28 human platelet antigens (HPAs) have been indentified on six polymorphic glycoproteins on the surface of platelets. Antibodies

  6. Virosome-mediated delivery of protein antigens to dendritic cells

    NARCIS (Netherlands)

    Bungener, L; Serre, K; Bijl, L; Leserman, L; Wilschut, J; Daemen, T; Machy, P

    2002-01-01

    Virosomes are reconstituted viral membranes in which protein can be encapsulated. Fusion-active virosomes, fusion-inactive virosomes and liposomes were used to study the conditions needed for delivery of encapsulated protein antigen ovalbumin (OVA) to dendritic cells (DCs) for MHC class I and 11

  7. Histological evaluation of the root apices of failed endodontic cases

    Directory of Open Access Journals (Sweden)

    Camilla Nicole Pecora

    2015-01-01

    Full Text Available Aim: The aim of this histological study was to evaluate the different causes of the failures of the endodontic treatment. Materials and Methods: Adult patients who were referred for endodontic surgery and for whom root-end resection was considered suitable were invited to participate in this study. The inclusion criteria were: (1 Patients aged between 18 and 65 years; (2 teeth with apical periodontitis that was diagnosed radiographically; (3 the tooth could not be adequately and better managed by root-canal retreatment; and (4 the crown of the tooth was adequately restored. One hundred root apices were surgically removed together with the periapical pathological tissue from 92 patients (56 males and 36 women.Histological sectioning was performed on calcified specimens that were embedded in a suitable medium. Results: The causes of endodontic failure identified through histological evaluation were as follows: Presence of bacterial and debris into canals (51%, apex transposition and overfilling (3%, presence of the isthmus (21%, bacterial colonization of root surface (2%, untreated canals (11%, and filling material outside root canal (4%. There was no significant difference between the distribution of teeth and the causes of endodontic failures (P = 0.32. Conclusions: The present study demonstrated that the most common cause of endodontic failure is the insufficient cleaning of the root canal system. It also explores the effectiveness of histological evaluation of the root apex following root-end resection in exploring the causes of endodontic failures.

  8. Apical canal sealing ability of calcium phosphate based cements.

    Science.gov (United States)

    Yoshikawa, M; Inamoto, T; Hakata, T; Toda, T

    1996-12-01

    The apical canal sealing ability of newly developed calcium phosphate cement (TDM-C), an equimolar mixture of tetracalcium and dicalcium phosphate dihydrate kneaded with McIlvain's buffer solution containing sodium carboxymethyl cellulose, was evaluated in a relative comparison with several recognized available products. The latter included BONETRIX (alpha-tricalcium phosphate mixed with citric acid and tannic acid), ARS (alpha-tricalcium phosphate and hydroxyapatite mixed with polyacrylic acid), and zine oxide eugenol sealer (ZOE). Canals of forty extracted human maxillary central incisors were prepared to the file size #70, divided into four groups, and obturated with each of the four above materials. Setting of all sealers took place either in 30% bovine serum solution or in phosphate buffered saline solution. Teeth were then decalcified and cleared to measure linear penetration of India ink from the apex into the canal. Results were statistically analyzed by Student's t-test. Surfaces of all materials, after setting, were also observed by scanning electron microscopy (SEM). We found that TDM-C, BONETRIX, and ZOE all showed excellent apical sealing ability whereas ARS was less effective in this regard. SEM analysis revealed micropores on the surface of every apatitic product, but ZOE was free of such porosity. It was concluded that TDM-C and BONETRIX may prove clinically useful as root canal sealant materials.

  9. Apical versus Basal Neurogenesis Directs Cortical Interneuron Subclass Fate.

    Science.gov (United States)

    Petros, Timothy J; Bultje, Ronald S; Ross, M Elizabeth; Fishell, Gord; Anderson, Stewart A

    2015-11-10

    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. Copyright © 2015 The Authors. Published by Elsevier Inc. All rights reserved.

  10. 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

  11. Unpolarized Release of Vaccinia Virus and HIV Antigen by Colchicine Treatment Enhances Intranasal HIV Antigen Expression and Mucosal Humoral Responses

    Science.gov (United States)

    Zhang, Yan; Yang, Jingyi; Bao, Rong; Chen, Yaoqing; Zhou, Dihan; He, Benxia; Zhong, Maohua; Li, Yaoming; Liu, Fang; Li, Qiaoli; Yang, Yi; Han, Chen; Sun, Ying; Cao, Yuan; Yan, Huimin

    2011-01-01

    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. PMID:21935396

  12. 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.

  13. Membranous nephropathy

    Science.gov (United States)

    ... check for hepatitis B, hepatitis C, and syphilis Complement levels Cryoglobulin test Treatment The goal of treatment ... not as helpful for people with membranous nephropathy. Medicines used treat membranous nephropathy include: Angiotensin-converting enzyme ( ...

  14. 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...

  15. Light Responsive Polymer Membranes: A Review

    Directory of Open Access Journals (Sweden)

    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.

  16. In vivo PTH provokes apical NHE3 and NaPi2 redistribution and Na-K-ATPase inhibition

    DEFF Research Database (Denmark)

    Zhang, Y; Norian, J M; Magyar, C E

    1999-01-01

    The aim of this study was to test the hypothesis that in vivo administration of parathyroid hormone (PTH) provokes diuresis/natriuresis through redistribution of proximal tubule apical sodium cotransporters (NHE3 and NaPi2) to internal stores and inhibition of basolateral Na-K-ATPase activity...... and to determine whether the same cellular signals drive the changes in apical and basolateral transporters. PTH-(1-34) (20 U), which couples to adenylate cyclase (AC), phospholipase C (PLC), and phospholipase A2 (PLA2), or [Nle8,18,Tyr34]PTH-(3-34) (10 U), which couples to PLC and PLA2 but not AC, were given...... to anesthetized rats as an intravenous bolus followed by low-dose infusion (1 U. kg-1. min-1 for 1 h). Renal cortex membranes were fractionated on sorbitol density gradients. PTH-(1-34) increased urinary cAMP excretion 3-fold, urine output (V) 2.0 +/- 0.1-fold, and lithium clearance (CLi) 2.8 +/- 0.3-fold...

  17. Ultrastructure of the harmful unarmored dinoflagellate Cochlodinium polykrikoides (Dinophyceae) with reference to the apical groove and Flagellar apparatus.

    Science.gov (United States)

    Iwataki, Mitsunori; Hansen, Gert; Moestrup, Øjvind; Matsuoka, Kazumi

    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 sulcal extension on the dorsal side of the epicone. The eyespot is located dorsally and composed of two layers of globules situated within the chloroplast. A narrow invagination of the plasma membrane is associated with the eyespot. The nuclear envelope has normal nuclear pores similar to other 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 microm and interconnected directly by a striated basal body connective and indirectly by microtubular and fibrous structures. Characteristic features of the flagellar apparatus are as follows: (1) a nuclear extension projects to the R1 (longitudinal microtubular root) and is connected to the root by thin fibrous material; (2) fibrillar structures are associated with the longitudinal and transverse flagellar canal; and (3) a striated ventral connective extends toward the posterior end of the cell along the longitudinal flagellar canal. We conclude, based on both morphological and molecular evidence, that Cochlodinium is only distantly related to Gymnodinium.

  18. Antigen entrapped in the escheriosomes leads to the generation of CD4(+) helper and CD8(+) cytotoxic T cell response.

    Science.gov (United States)

    Syed, Faisal M; Khan, Masood A; Nasti, Tahseen H; Ahmad, Nadeem; Mohammad, Owais

    2003-06-02

    In previous study, we demonstrated the potential of Escherichia coli (E. coli) lipid liposomes (escheriosomes) to undergo membrane-membrane fusion with cytoplasmic membrane of the target cells including professional antigen presenting cells. Our present study demonstrates that antigen encapsulated in escheriosomes could be successfully delivered simultaneously to the cytosolic as well as endosomal processing pathways of antigen presenting cells, leading to the generation of both CD4(+) T-helper and CD8(+) cytotoxic T cell response. In contrast, encapsulation of same antigen in egg phosphatidyl-choline (egg PC) liposomes, just like antigen-incomplete Freund's adjuvant (IFA) complex, has inefficient access to the cytosolic pathway of MHC I-dependent antigen presentation and failed to generate antigen-specific CD8(+) cytotoxic T cell response. However, both egg PC liposomes as well as escheriosomes-encapsulated antigen elicited strong humoral immune response in immunized animals but antibody titre was significantly higher in the group of animals immunized with escheriosomes-encapsulated antigen. These results imply usage of liposome-based adjuvant as potential candidate vaccine capable of eliciting both cell-mediated as well as humoral immune responses. Furthermore, antigen entrapped in escheriosomes stimulates antigen-specific CD4(+) T cell proliferation and also enhances the level of IL-2, IFN-gamma and IL-4 in the immunized animals.

  19. The C-terminal hypervariable domain targets Aradopsis ROP9 to the invaginated pollen tube plasma membrane

    Science.gov (United States)

    Rop9 is a small GTPase of the Type II class, whereas the often studied type I Rops play roles during pollen tube growth. In pollen, Rop9 is located at the invaginated plasma membrane that surrounds the sperm cells, whereas type I Rops are located at the apical membrane of the pollen tube. The C-ter...

  20. A monkey antigen crossreacting with carcinoembryonic antigen, CEA.

    Science.gov (United States)

    Engvall, E.; Vuento, M.; Ruoslahti, E.

    1976-01-01

    Normal monkey tissues were found to contain an antigen which crossreacts immunologically with the carcinoembryonic antigen (CEA) of the human digestive tract. The monkey antigen reacted with complete or partial identity to the normal crossreacting antigen (NCA) in humans when tested in immunodiffusion against anti-CEA or anti-NCA. Extracts of monkey tissues inhibited in radioimmunoassays measuring human NCA. It is possible that monkey foetuses and colonic tumours contain CEA. Images Fig. 1 PMID:823952

  1. Antigen smuggling in tuberculosis.

    Science.gov (United States)

    Hudrisier, Denis; Neyrolles, Olivier

    2014-06-11

    The importance of CD4 T lymphocytes in immunity to M. tuberculosis is well established; however, how dendritic cells activate T cells in vivo remains obscure. In this issue of Cell Host & Microbe, Srivastava and Ernst (2014) report a mechanism of antigen transfer for efficient activation of antimycobacterial T cells. Copyright © 2014 Elsevier Inc. All rights reserved.

  2. CD antigens 2001

    Czech Academy of Sciences Publication Activity Database

    Mason, D.; Andre, P.; Bensussan, A.; Buckley, C.; Civin, C.; Clark, E.; de Haas, M.; Goyert, S.; Hadam, M.; Hart, D.; Hořejší, Václav; Meuer, S.; Morrissey, J.; Schwartz-Albiez, R.; Shaw, S.; Simmons, D.; Uguccioni, M.; van der Schoot, E.; Vivier, E.; Zola, H.

    2001-01-01

    Roč. 58, č. 6 (2001), s. 425-430 ISSN 0001-2815. [Conference on Human leucocyte differentiation antigens /7./. Harrogate, 20.06.2000-25.06.2000] Institutional research plan: CEZ:AV0Z5052915 Keywords : CD molecules, HLDA Subject RIV: EC - Immunology Impact factor: 2.864, year: 2001

  3. CD antigens 2002

    Czech Academy of Sciences Publication Activity Database

    Mason, D.; Andre, P.; Bensussan, A.; Buckley, C.; Civin, C.; Clark, E.; de Haas, M.; Goyert, S.; Hadam, M.; Hart, D.; Hořejší, Václav; Meuer, S.; Morrissey, J.; Schwartz-Albiez, R.; Shaw, S.; Simmons, D.; Uguccioni, M.; van der Schoot, E.; Vivier, E.; Zola, H.

    2002-01-01

    Roč. 99, č. 10 (2002), s. 3877-3880 ISSN 0006-4971. [Conference on Human leucocyte differentiation antigens /7./. Harrogate, 20.06.2000-25.06.2000] Institutional research plan: CEZ:AV0Z5052915 Keywords : CD molecules, HLDA Subject RIV: EC - Immunology Impact factor: 9.631, year: 2002

  4. CD antigens 2001

    Czech Academy of Sciences Publication Activity Database

    Mason, D.; Andre, P.; Bensussan, A.; Buckley, C.; Civin, C.; Clark, E.; de Haas, M.; Goyert, S.; Hadam, M.; Hart, D.; Hořejší, Václav; Meuer, S.; Morrissey, J.; Schwartz-Albiez, R.; Shaw, S.; Simmons, D.; Uguccioni, M.; van der Schoot, E.; Vivier, E.; Zola, H.

    2002-01-01

    Roč. 168, č. 5 (2002), s. 2083-2086 ISSN 0022-1767. [Conference on Human leucocyte differentiation antigens /7./. Harrogate, 20.06.2000-25.06.2000] Institutional research plan: CEZ:AV0Z5052915 Keywords : CD molecules, HLDA Subject RIV: EC - Immunology Impact factor: 7.014, year: 2002

  5. CD antigens 2001

    Czech Academy of Sciences Publication Activity Database

    Mason, D.; Andre, P.; Bensussan, A.; Buckley, C.; Civin, C.; Clark, E.; de Haas, M.; Goyert, S.; Hadam, M.; Hart, D.; Hořejší, Václav; Meuer, S.; Morrissey, J.; Schwartz-Albiez, R.; Shaw, S.; Simmons, D.; Uguccioni, M.; van der Schoot, E.; Vivier, E.; Zola, H.

    2001-01-01

    Roč. 103, č. 4 (2001), s. 401-406 ISSN 0019-2805 R&D Projects: GA ČR GA310/99/0349 Institutional research plan: CEZ:AV0Z5052915 Keywords : antigen * CD * leukocyte Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 2.656, year: 2001

  6. CD antigens 2001

    Czech Academy of Sciences Publication Activity Database

    Mason, D.; Andre, P.; Bensussan, A.; Buckley, C.; Civin, C.; Clark, E.; de Haas, M.; Goyert, S.; Hadam, M.; Hart, D.; Hořejší, Václav; Meuer, S.; Morrissey, J.; Schwartz-Albiez, R.; Shaw, S.; Simmons, D.; Uguccioni, M.; van der Schoot, E.; Vivier, E.; Zola, H.

    2001-01-01

    Roč. 19, č. 6 (2001), s. 556-562 ISSN 1066-5099 R&D Projects: GA AV ČR IAA7052904 Institutional research plan: CEZ:AV0Z5052915 Keywords : CD * leukocyte antigens Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 2.689, year: 2001

  7. CD antigens 2001

    Czech Academy of Sciences Publication Activity Database

    Mason, D.; Andre, P.; Bensussan, A.; Buckley, C.; Civin, C.; Clark, E.; de Haas, M.; Goyert, S.; Hadam, M.; Hart, D.; Hořejší, Václav; Meuer, S.; Morrissey, J.; Schwartz-Albiez, R.; Shaw, S.; Simmons, D.; Uguccioni, M.; van der Schoot, E.; Vivier, E.; Zola, H.

    2001-01-01

    Roč. 31, č. 10 (2001), s. 2841-2847 ISSN 0014-2980 R&D Projects: GA AV ČR IAA7052904 Keywords : CD * leukocyte antigens Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 4.990, year: 2001

  8. CD antigens 2001

    Czech Academy of Sciences Publication Activity Database

    Mason, D.; Andre, P.; Bensussan, A.; Buckley, C.; Civin, C.; Clark, E.; de Haas, M.; Goyert, S.; Hadam, M.; Hart, D.; Hořejší, Václav; Meuer, S.; Morrissey, J.; Schwartz-Albiez, R.; Shaw, S.; Simmons, D.; Uguccioni, M.; van der Schoot, E.; Vivier, E.; Zola, H.

    2001-01-01

    Roč. 211, č. 2 (2001), s. 81-85 ISSN 0008-8749 R&D Projects: GA ČR GA310/99/0349 Institutional research plan: CEZ:AV0Z5052915 Keywords : antigen * CD * leukocyte Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 2.604, year: 2001

  9. CD antigens 2001

    Czech Academy of Sciences Publication Activity Database

    Mason, D.; Andre, P.; Bensussan, A.; Buckley, C.; Civin, C.; Clark, E.; de Haas, M.; Goyert, S.; Hadam, M.; Hart, D.; Hořejší, Václav; Meuer, S.; Morrissey, J.; Schwartz-Albiez, R.; Shaw, S.; Simmons, D.; Uguccioni, M.; van der Schoot, E.; Vivier, E.; Zola, H.

    2002-01-01

    Roč. 15, č. 1 (2002), s. 71-76 ISSN 0893-3952. [Conference on Human leucocyte differentiation antigens /7./. Harrogate, 20.06.2000-25.06.2000] Institutional research plan: CEZ:AV0Z5052915 Keywords : CD molecules, HLDA Subject RIV: EC - Immunology Impact factor: 3.821, year: 2002

  10. CD antigens 2001

    Czech Academy of Sciences Publication Activity Database

    Mason, D.; Andre, P.; Bensussan, A.; Buckley, C.; Civin, C.; Clark, E.; de Haas, M.; Goyert, S.; Hadam, M.; Hart, D.; Hořejší, Václav; Meuer, S.; Morrissey, J.; Schwartz-Albiez, R.; Shaw, S.; Simmons, D.; Uguccioni, M.; van der Schoot, E.; Vivier, E.; Zola, H.

    2001-01-01

    Roč. 70, č. 5 (2001), s. 685-690 ISSN 0741-5400 R&D Projects: GA AV ČR IAA7052904 Institutional research plan: CEZ:AV0Z5052915 Keywords : CD * leukocyte antigens Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 4.516, year: 2001

  11. CD antigens 2001

    Czech Academy of Sciences Publication Activity Database

    Mason, D.; Andre, P.; Bensussan, A.; Buckley, C.; Civin, C.; Clark, E.; de Haas, M.; Goyert, S.; Hadam, M.; Hart, D.; Hořejší, Václav; Meuer, S.; Morrissey, J.; Schwartz-Albiez, R.; Shaw, S.; Simmons, D.; Uguccioni, M.; van der Schoot, E.; Vivier, E.; Zola, H.

    2001-01-01

    Roč. 13, č. 9 (2001), s. 1095-1098 ISSN 0953-8178 R&D Projects: GA ČR GA310/99/0349 Institutional research plan: CEZ:AV0Z5052915 Keywords : antigen * CD * leukocyte Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 3.611, year: 2001

  12. β-endorphin antigen

    International Nuclear Information System (INIS)

    1981-01-01

    This invention relates to the production of antigens comprising β-endorphin, βsub(h)-endorphin, or βsub(c)-endorphin, in covalent conjugation with human gammaglobulin as immunogenic carrier material, and an antibody having the property of specifically binding β-endorphin or fragments thereof, containing the (6-15) residue sequence. (U.K.)

  13. Disappearance of paternal histocompatibility antigens from hybrid mouse blastocyst at the time of implantation.

    Science.gov (United States)

    Leclipteux, T; Remacle, J

    1983-07-04

    Products of the major histocompatibility complex (H-2) are important in allograph rejection. In view of the close relationship between mother and foetus, we can consider the latter as an allograph which is however not rejected by an immunological reaction. We studied the presence of H-2 antigens on embryo membranes at the time of implantation, by immunochemical labeling using gold particles coupled with protein A. Results showed that the expression of H-2 antigens is different before and after implantation. It seems that after implantation, H-2 antigens disappear from trophoblastic membranes. This could explain the absence of immunological reaction of the mother against the foetus.

  14. 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.

  15. Differential subcellular membrane recruitment of Src may specify its downstream signalling

    International Nuclear Information System (INIS)

    Diesbach, Philippe de; Medts, Thierry; Carpentier, Sarah; D'Auria, Ludovic; Van Der Smissen, Patrick; Platek, Anna; Mettlen, Marcel; Caplanusi, Adrian; Hove, Marie-France van den; Tyteca, Donatienne; Courtoy, Pierre J.

    2008-01-01

    Most Src family members are diacylated and constitutively associate with membrane 'lipid rafts' that coordinate signalling. Whether the monoacylated Src, frequently hyperactive in carcinomas, also localizes at 'rafts' remains controversial. Using polarized MDCK cells expressing the thermosensitive v-Src/tsLA31 variant, we here addressed how Src tyrosine-kinase activation may impact on its (i) membrane recruitment, in particular to 'lipid rafts'; (ii) subcellular localization; and (iii) signalling. The kinetics of Src-kinase thermoactivation correlated with its recruitment from the cytosol to sedimentable membranes where Src largely resisted solubilisation by non-ionic detergents at 4 deg. C and floated into sucrose density gradients like caveolin-1 and flotillin-2, i.e. 'lipid rafts'. By immunofluorescence, activated Src showed a dual localization, at apical endosomes/macropinosomes and at the apical plasma membrane. The plasma membrane Src pool did not colocalize with caveolin-1 and flotillin-2, but extensively overlapped GM1 labelling by cholera toxin. Severe (∼ 70%) cholesterol extraction with methyl-β-cyclodextrin (MβCD) did not abolish 'rafts' floatation, but strongly decreased Src association with floating 'rafts' and abolished its localization at the apical plasma membrane. Src activation independently activated first the MAP-kinase - ERK1/2 pathway, then the PI3-kinase - Akt pathway. MAP-kinase - ERK1/2 activation was insensitive to MβCD, which suppressed Akt phosphorylation and apical endocytosis induced by Src, both depending on the PI3-kinase pathway. We therefore suggest that activated Src is recruited at two membrane compartments, allowing differential signalling, first via ERK1/2 at 'non-raft' domains on endosomes, then via PI3-kinase-Akt on a distinct set of 'rafts' at the apical plasma membrane. Whether this model is applicable to c-Src remains to be examined

  16. Influence of Different Apical Preparations on Root Canal Cleanliness in Human Molars: a SEM Study.

    Science.gov (United States)

    Plotino, Gianluca; Grande, Nicola M; Tocci, Luigi; Testarelli, Luca; Gambarini, Gianluca

    2014-04-01

    To compare the influence of type and dimensions of the apical preparation on the cleanliness of the apical area in molars. A total of 120 root canals (MB and DB root canals from 30 maxillary molars and mesial root canals from 30 mandibular molars) were instrumented with Mtwo NiTi rotary instruments to a size 25/0.06 taper and were equally divided into three different experimental groups depending on the subsequently apical root canal preparation: Group 1: no further apical preparation, Group 2: apical preparation with Mtwo files to a size 40/0.04 taper, Group 3: apical preparation with Mtwo Apical Files. All root canals were observed through scanning electron microscopy (SEM). Presence of superficial debris and smear layer was evaluated using a score system. Data were statistically analysed using the Kruskal-Wallis and Bonferroni tests with a level of significance set at P 0.05), while at the apical level, there was a significant difference for both residual debris and presence of smear layer between Group 1 and both Group 2 (P = 0.003 and P = 0.014) and 3 (P = 0.012 and P = 0.021), while no difference was present between Group 2 and Group 3 (P = 0.871 and P = 0.923). Cleanliness of the apical third in terms of debris and smear layer was statistically better when an apical preparation was performed to a size 40/0.04 taper or with the use of the Mtwo Apical Files.

  17. Influence of Different Apical Preparations on Root Canal Cleanliness in Human Molars: a SEM Study

    Directory of Open Access Journals (Sweden)

    Gianluca Plotino

    2014-07-01

    Full Text Available Objectives: To compare the influence of type and dimensions of the apical preparation on the cleanliness of the apical area in molars. Material and Methods: A total of 120 root canals (MB and DB root canals from 30 maxillary molars and mesial root canals from 30 mandibular molars were instrumented with Mtwo NiTi rotary instruments to a size 25/0.06 taper and were equally divided into three different experimental groups depending on the subsequently apical root canal preparation: Group 1: no further apical preparation, Group 2: apical preparation with Mtwo files to a size 40/0.04 taper, Group 3: apical preparation with Mtwo Apical Files. All root canals were observed through scanning electron microscopy (SEM. Presence of superficial debris and smear layer was evaluated using a score system. Data were statistically analysed using the Kruskal-Wallis and Bonferroni tests with a level of significance set at P 0.05, while at the apical level, there was a significant difference for both residual debris and presence of smear layer between Group 1 and both Group 2 (P = 0.003 and P = 0.014 and 3 (P = 0.012 and P = 0.021, while no difference was present between Group 2 and Group 3 (P = 0.871 and P = 0.923. Conclusions: Cleanliness of the apical third in terms of debris and smear layer was statistically better when an apical preparation was performed to a size 40/0.04 taper or with the use of the Mtwo Apical Files.

  18. Bone Density and Dental External Apical Root Resorption.

    Science.gov (United States)

    Iglesias-Linares, Alejandro; Morford, Lorri Ann; Hartsfield, James Kennedy

    2016-12-01

    When orthodontic patients desire shorter treatment times with aesthetic results and long-term stability, it is important for the orthodontist to understand the potential limitations and problems that may arise during standard and/or technology-assisted accelerated treatment. Bone density plays an important role in facilitating orthodontic tooth movement (OTM), such that reductions in bone density can significantly increase movement velocity. Lifestyle, genetic background, environmental factors, and disease status all can influence a patients' overall health and bone density. In some individuals, these factors may create specific conditions that influence systemic-wide bone metabolism. Both genetic variation and the onset of a bone-related disease can influence systemic bone density and local bone density, such as observed in the mandible and maxilla. These types of localized density changes can affect the rate of OTM and may also influence the risk of unwanted outcomes, i.e., the occurrence of dental external apical root resorption (EARR).

  19. 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)

  20. Traumatic bone cyst suggestive of large apical periodontitis.

    Science.gov (United States)

    Rodrigues, Cleomar Donizeth; Estrela, Carlos

    2008-04-01

    This case report shows the importance of establishing the correct diagnosis to provide the appropriate treatment options The traumatic bone cyst is a pseudocyst, usually asymptomatic and found by a routine radiographic examination. Unicystic radiolucency is almost always observed, which can involve the periradicular area of teeth, simulating an inflammatory lesion of endodontic origin. Differential diagnosis should include other pathologies, such as odontogenic keratocyst, central giant cell granuloma, and unicystic ameloblastoma. Its etiology and pathogenesis are not yet definitely established. In the present study, after review of the medical and dental histories and clinical and radiographic examination of teeth #24-27 (pulpal vitality test showed positive), the primary diagnosis was traumatic bone cyst. The planning was excisional biopsy. After surgical exploration, only one small blood clot was observed in the intraosseous socket, which was carefully curetted and filled with blood. A clinical and radiographic examination after 6 months showed apical formation and pulpal vitality preserved.

  1. Geometry of shoot apical dome and distribution of growth rates

    Directory of Open Access Journals (Sweden)

    Jerzy Nakielski

    2014-01-01

    Full Text Available The distribution of the relative elementary rate of growth (RERG in apical domes of various shapes and patterns of displacement lines can be analytically examined. The geometry of these domes may be described by parabolas of n-th order, the variant of the distribution of linear growth rate should be established along any displacement line (e.g. along the axis and then the RERG can be studied as the function depending on the position coordinates and the parameter n. Such investigations of several aplical domes of various shapes have been performed. The results confirm the occurrence of the minimum of relative, elementary growth rate (in volume in the subapical region of the dome independently of the type of geometry (n parabola order.

  2. The sodium-bicarbonate cotransporter NBCe2 (slc4a5) expressed in human renal proximal tubules shows increased apical expression under high-salt conditions.

    Science.gov (United States)

    Gildea, John J; Xu, Peng; Carlson, Julia M; Gaglione, Robert T; Bigler Wang, Dora; Kemp, Brandon A; Reyes, Camellia M; McGrath, Helen E; Carey, Robert M; Jose, Pedro A; Felder, Robin A

    2015-12-01

    The electrogenic sodium bicarbonate cotransporter (NBCe2) is encoded by SLC4A5, variants of which have been associated with salt sensitivity of blood pressure, which affects 25% of the adult population. NBCe2 is thought to mediate sodium bicarbonate cotransport primarily in the renal collecting duct, but NBCe2 mRNA is also found in the rodent renal proximal tubule (RPT). The protein expression or function of NBCe2 has not been demonstrated in the human RPT. We validated an NBCe2 antibody by shRNA and Western blot analysis, as well as overexpression of an epitope-tagged NBCe2 construct in both RPT cells (RPTCs) and human embryonic kidney 293 (HEK293) cells. Using this validated NBCe2 antibody, we found NBCe2 protein expression in the RPT of fresh and frozen human kidney slices, RPTCs isolated from human urine, and isolated RPTC apical membrane. Under basal conditions, NBCe2 was primarily found in the Golgi, while NBCe1 was primarily found at the basolateral membrane. Following an acute short-term increase in intracellular sodium, NBCe2 expression was increased at the apical membrane in cultured slices of human kidney and polarized, immortalized RPTCs. Sodium bicarbonate transport was increased by monensin and overexpression of NBCe2, decreased by NBCe2 shRNA, but not by NBCe1 shRNA, and blocked by 2,2'-(1,2-ethenediyl)bis[5-isothiocyanato-benzenesulfonic acid]. NBCe2 could be important in apical sodium and bicarbonate cotransport under high-salt conditions; the implication of the ex vivo studies to the in vivo situation when salt intake is increased remains unclear. Therefore, future studies will examine the role of NBCe2 in mediating increased renal sodium transport in humans whose blood pressures are elevated by an increase in sodium intake. Copyright © 2015 the American Physiological Society.

  3. A network of spectrin and plectin surrounds the actin cuffs of apical tubulobulbar complexes in the rat.

    Science.gov (United States)

    Aristaeus de Asis, Marc; Pires, Manuel; Lyon, Kevin; Vogl, A Wayne

    2013-07-01

    Tubulobulbar complexes (TBCs) are actin-related endocytic structures that internalize intercellular junctions in the seminiferous epithelium. The structures consist of elongate tubular projections of the attached plasma membranes of two adjacent cells that project into Sertoli cells. This double membrane core is cuffed by a dentritic actin network and is capped at its end by a clathrin-coated pit. Here we explore the possibility that elements of the spectrin cytoskeleton are associated with clusters of tubulobulbar complexes that develop at adhesion junctions between late spermatids and Sertoli cells at the apex of the epithelium, and extend what is known about the distribution of plectin at the sites. Cryo-sections of perfusion-fixed testes and apical processes of Sertoli cells mechanically dissociated from perfusion-fixed testes were probed for spectrin, EPB41, and actin and analyzed using conventional fluorescence microscopy and confocal microscopy. Data sets from confocal microscopy were analyzed further in three-dimensional reconstructions using computer software. Additional apical Sertoli cell processes were probed for plectin and analyzed using conventional fluorescence microscopy. Antibodies generated against elements of the spectrin cytoskeleton react with material around and between the actin cuffs of tubulobulbar complexes, but appear excluded from the actin cuffs themselves. A similar staining pattern occurs with a probe for plectin. Immunoelectron microscopy confirmed the staining patterns observed by fluourescence microscopy. Based on our results, we suggest that a network of spectrin and plectin forms a scaffold around tubulobulbar complexes that may provide support for the actin network that cuffs each complex and also link adjacent complexes together.

  4. An unusual ST-segment elevation: apical hypertrophic cardiomyopathy shows the ace up its sleeve.

    Science.gov (United States)

    de Santis, Francesco; Pergolini, Amedeo; Zampi, Giordano; Pero, Gaetano; Pino, Paolo Giuseppe; Minardi, Giovanni

    2013-01-01

    Apical hypertrophic cardiomyopathy is part of the broad clinical and morphologic spectrum of hypertrophic cardiomyopathy. We report a patient with electrocardiographic abnormalities in whom acute coronary syndrome was excluded and apical hypertrophic cardiomyopathy was demonstrated by careful differential diagnosis. Copyright © 2012 Sociedade Portuguesa de Cardiologia. Published by Elsevier España. All rights reserved.

  5. Nonsurgical root canal therapy of large cyst-like inflammatory periapical lesions and inflammatory apical cysts.

    Science.gov (United States)

    Lin, Louis M; Ricucci, Domenico; Lin, Jarshen; Rosenberg, Paul A

    2009-05-01

    It is a general belief that large cyst-like periapical lesions and apical true cysts caused by root canal infection are less likely to heal after nonsurgical root canal therapy. Nevertheless, there is no direct evidence to support this assumption. A large cyst-like periapical lesion or an apical true cyst is formed within an area of apical periodontitis and cannot form by itself. Therefore, both large cyst-like periapical lesions and apical true cysts are of inflammatory and not of neoplastic origin. Apical periodontitis lesions, regardless of whether they are granulomas, abscesses, or cysts, fail to heal after nonsurgical root canal therapy for the same reason, intraradicular and/or extraradicular infection. If the microbial etiology of large cyst-like periapical lesions and inflammatory apical true cysts in the root canal is removed by nonsurgical root canal therapy, the lesions might regress by the mechanism of apoptosis in a manner similar to the resolution of inflammatory apical pocket cysts. To achieve satisfactory periapical wound healing, surgical removal of an apical true cyst must include elimination of root canal infection.

  6. 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...

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

    DEFF Research Database (Denmark)

    von Arx, Thomas; Peñarrocha, Miguel; Jensen, Simon 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...

  8. Apical bud toughness tests and tree sway movements to examine crown abrasion: preliminary results

    Science.gov (United States)

    Tyler Brannon; Wayne Clatterbuck

    2012-01-01

    Apical bud toughness differences were examined for several species to determine if crown abrasion affects shoot growth of determinate and indeterminate species during stand development. Determinate buds will set and harden after initial shoot elongation in the spring, while the indeterminate shoots form leaves from the apical meristem continuously based on the...

  9. Plasma membranes from insect midgut cells

    Directory of Open Access Journals (Sweden)

    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

  10. Glyphosate effects on the gene expression of the apical bud in soybean (Glycine max).

    Science.gov (United States)

    Jiang, Ling-Xue; Jin, Long-Guo; Guo, Yong; Tao, Bo; Qiu, Li-Juan

    2013-08-09

    Glyphosate is a broad spectrum, non-selective herbicide which has been widely used for weed control. Much work has focused on elucidating the high accumulation of glyphosate in shoot apical bud (shoot apex). However, to date little is known about the molecular mechanisms of the sensitivity of shoot apical bud to glyphosate. Global gene expression profiling of the soybean apical bud response to glyphosate treatment was performed in this study. The results revealed that the glyphosate inhibited tryptophan biosynthesis of the shikimic acid pathway in the soybean apical bud, which was the target site of glyphosate. Glyphosate inhibited the expression of most of the target herbicide site genes. The promoter sequence analysis of key target genes revealed that light responsive elements were important regulators in glyphosate induction. These results will facilitate further studies of cloning genes and molecular mechanisms of glyphosate on soybean shoot apical bud. Copyright © 2013 Elsevier Inc. All rights reserved.

  11. Endodontic management of open apex using Biodentine as a novel apical matrix.

    Science.gov (United States)

    Khetarpal, Ambica; Chaudhary, Sarika; Talwar, Sangeeta; Verma, Mahesh

    2014-01-01

    Endodontic management of open apex using Biodentine as an apical matrix. Summary : An immature tooth with pulpal necrosis and periapical pathology imposes a great difficulty to the endodontist. Endodontic treatment options for such teeth consist of conventional apexification procedure with and without apical barriers. Biodentine™ is new calcium silicate based cement that exhibits physical and chemical properties similar to those described for certain Portland cement derivatives. This article demonstrates the use of the newer material, Biodentine as an apical matrix barrier in root end apexification procedure. This case reports present apexification and successful healing with the use of Biodentine as an apical barrier matrix. Conclusion : Apexification in one step using an apical plug of Biodentine can be considered a predictable treatment and may be an alternative to mineral trioxide aggregate apexification.

  12. Laparoscopic pectopexy: initial experience of single center with a new technique for apical prolapse surgery

    Directory of Open Access Journals (Sweden)

    Ahmet Kale

    Full Text Available ABSTRACT Objective: To share our first experience with laparoscopic pectopexy, a new technique for apical prolapse surgery, and to evaluate the feasibility of this technique. Materials and Methods: Seven patients with apical prolapse underwent surgery with laparoscopic pectopexy. The lateral parts of the iliopectineal ligament were used for a bilateral mesh fixation of the descended structures. The medical records of the patients were reviewed, and the short-term clinical outcomes were analyzed. Results: The laparoscopic pectopexy procedures were successfully performed, without intraoperative and postoperative complications. De novo apical prolapse, de novo urgency, de novo constipation, stress urinary incontinence, anterior and lateral defect cystoceles, and rectoceles did not occur in any of the patients during a 6-month follow-up period. Conclusion: Although laparoscopic sacrocolpopexy has shown excellent anatomical and functional long-term results, laparoscopic pectopexy offers a feasible, safe, and comfortable alternative for apical prolapse surgery. Pectopexy may increase a surgeon's technical perspective for apical prolapse surgery.

  13. Cytoskeletal Components of an Invasion Machine—The Apical Complex of Toxoplasma gondii

    Science.gov (United States)

    Hu, Ke; Johnson, Jeff; Florens, Laurence; Fraunholz, Martin; Suravajjala, Sapna; DiLullo, Camille; Yates, John; Roos, David S; Murray, John M

    2006-01-01

    The apical complex of Toxoplasma gondii is widely believed to serve essential functions in both invasion of its host cells (including human cells), and in replication of the parasite. The understanding of apical complex function, the basis for its novel structure, and the mechanism for its motility are greatly impeded by lack of knowledge of its molecular composition. We have partially purified the conoid/apical complex, identified ~200 proteins that represent 70% of its cytoskeletal protein components, characterized seven novel proteins, and determined the sequence of recruitment of five of these proteins into the cytoskeleton during cell division. Our results provide new markers for the different subcompartments within the apical complex, and revealed previously unknown cellular compartments, which facilitate our understanding of how the invasion machinery is built. Surprisingly, the extreme apical and extreme basal structures of this highly polarized cell originate in the same location and at the same time very early during parasite replication. PMID:16518471

  14. 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

  15. Human platelet antigens - 2013.

    Science.gov (United States)

    Curtis, B R; McFarland, J G

    2014-02-01

    To date, 33 human platelet alloantigens (HPAs) have been identified on six functionally important platelet glycoprotein (GP) complexes and have been implicated in alloimmune platelet disorders including foetal and neonatal alloimmune thrombocytopenia (FNAIT), posttransfusion purpura (PTP) and multitransfusion platelet refractoriness (MPR). The greatest number of recognized HPA (20 of 33) resides on the GPIIb/IIIa complex, which serves as the receptor for ligands important in mediating haemostasis and inflammation. These include HPA-1a, the most commonly implicated HPA in FNAIT and PTP in Caucasian populations. Other platelet GP complexes, GPIb/V/IX, GPIa/IIa and CD109, express the remaining 13 HPAs. Of the recognized HPAs, 12 occur as six serologically and genetically defined biallelic 'systems' where the -a form designates the higher frequency allele and the -b form, the lower. Twenty-one other HPAs are low-frequency or rare antigens for which postulated higher frequency -a alleles have not yet been identified as antibody specificities. In addition to the HPA markers, platelets also express ABO and human leucocyte antigen (HLA) antigens; antibodies directed at the former are occasionally important in FNAIT, and to the latter, in MPR. © 2013 International Society of Blood Transfusion.

  16. Pro-oxidant status and matrix metalloproteinases in apical lesions and gingival crevicular fluid as potential biomarkers for asymptomatic apical periodontitis and endodontic treatment response

    Directory of Open Access Journals (Sweden)

    Dezerega Andrea

    2012-03-01

    Full Text Available Abstract Background Oxidative stress and matrix metalloproteinases -9 and -2 are involved in periodontal breakdown, whereas gingival crevicular fluid has been reported to reflect apical status. The aim of this study was to characterize oxidant balance and activity levels of MMP -2 and -9 in apical lesions and healthy periodontal ligament; and second, to determine whether potential changes in oxidant balance were reflected in gingival crevicular fluid from asymptomatic apical periodontitis (AAP-affected teeth at baseline and after endodontic treatment. Methods Patients with clinical diagnosis of AAP and healthy volunteers having indication of tooth extraction were recruited. Apical lesions and healthy periodontal ligaments, respectively, were homogenized or processed to obtain histological tissue sections. Matrix metalloproteinase -9 and -2 levels and/or activity were analyzed by Immunowestern blot, zymography and consecutive densitometric analysis, and their tissue localization was confirmed by immunohistochemistry. A second group of patients with AAP and indication of endodontic treatment was recruited. Gingival crevicular fluid was extracted from AAP-affected teeth at baseline, after endodontic treatment and healthy contralateral teeth. Total oxidant and antioxidant status were determined in homogenized tissue and GCF samples. Statistical analysis was performed using STATA v10 software with unpaired t test, Mann-Whitney test and Spearman's correlation. Results Activity of MMP-2 and MMP-9 along with oxidant status were higher in apical lesions (p Conclusions Apical lesions display an oxidant imbalance along with increased activity of matrix metalloproteinase-2 and -9 and might contribute to AAP progression. Oxidant imbalance can also be reflected in GCF from AAP-affected teeth and was restored to normal levels after conservative endodontic treatment. These mediators might be useful as potential biomarkers for chair-side complementary diagnostic

  17. Intraoperative cervix location and apical support stiffness in women with and without pelvic organ prolapse.

    Science.gov (United States)

    Swenson, Carolyn W; Smith, Tovia M; Luo, Jiajia; Kolenic, Giselle E; Ashton-Miller, James A; DeLancey, John O

    2017-02-01

    It is unknown how initial cervix location and cervical support resistance to traction, which we term "apical support stiffness," compare in women with different patterns of pelvic organ support. Defining a normal range of apical support stiffness is important to better understand the pathophysiology of apical support loss. The aims of our study were to determine whether: (1) women with normal apical support on clinic Pelvic Organ Prolapse Quantification, but with vaginal wall prolapse (cystocele and/or rectocele), have the same intraoperative cervix location and apical support stiffness as women with normal pelvic support; and (2) all women with apical prolapse have abnormal intraoperative cervix location and apical support stiffness. A third objective was to identify clinical and biomechanical factors independently associated with clinic Pelvic Organ Prolapse Quantification point C. We conducted an observational study of women with a full spectrum of pelvic organ support scheduled to undergo gynecologic surgery. All women underwent a preoperative clinic examination, including Pelvic Organ Prolapse Quantification. Cervix starting location and the resistance (stiffness) of its supports to being moved steadily in the direction of a traction force that increased from 0-18 N was measured intraoperatively using a computer-controlled servoactuator device. Women were divided into 3 groups for analysis according to their pelvic support as classified using the clinic Pelvic Organ Prolapse Quantification: (1) "normal/normal" was women with normal apical (C -5 cm and Ba and/or Bp ≥ 0 cm). Demographics, intraoperative cervix locations, and apical support stiffness values were then compared. Normal range of cervix location during clinic examination and operative testing was defined by the total range of values observed in the normal/normal group. The proportion of women in each group with cervix locations within and outside the normal range was determined. Linear regression

  18. Membrane paradigm

    International Nuclear Information System (INIS)

    Price, R.H.; Thorne, K.S.

    1986-01-01

    The membrane paradigm is a modified frozen star approach to modeling black holes, with particles and fields assuming a complex, static, boundary-layer type structure (membrane) near the event horizon. The membrane has no effects on the present or future evolution of particles and fields above itself. The mathematical representation is a combination of a formalism containing terms for the shear and bulk viscosity, surface pressure, momentum, temperature, entropy, etc., of the horizon and the 3+1 formalism. The latter model considers a family of three-dimensional spacelike hypersurfaces in one-dimensional time. The membrane model considers a magnetic field threading the hole and undergoing torque from the hole rotation. The field is cleaned by the horizon and distributed over the horizon so that ohmic dissipation is minimized. The membrane paradigm is invalid inside the horizon, but is useful for theoretically probing the properties of slowly evolving black holes

  19. Membrane processes

    Science.gov (United States)

    Staszak, Katarzyna

    2017-11-01

    The membrane processes have played important role in the industrial separation process. These technologies can be found in all industrial areas such as food, beverages, metallurgy, pulp and paper, textile, pharmaceutical, automotive, biotechnology and chemical industry, as well as in water treatment for domestic and industrial application. Although these processes are known since twentieth century, there are still many studies that focus on the testing of new membranes' materials and determining of conditions for optimal selectivity, i. e. the optimum transmembrane pressure (TMP) or permeate flux to minimize fouling. Moreover the researchers proposed some calculation methods to predict the membrane processes properties. In this article, the laboratory scale experiments of membrane separation techniques, as well their validation by calculation methods are presented. Because membrane is the "heart" of the process, experimental and computational methods for its characterization are also described.

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

    International Nuclear Information System (INIS)

    Ezoddini Ardakani, F.; Zangouie Booshehri, M.; Behniafar, B.

    2011-01-01

    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.

  1. 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.

  2. The Apical Complex Provides a Regulated Gateway for Secretion of Invasion Factors in Toxoplasma

    Science.gov (United States)

    Katris, Nicholas J.; van Dooren, Giel G.; McMillan, Paul J.; Hanssen, Eric; Tilley, Leann; Waller, Ross F.

    2014-01-01

    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. PMID:24743791

  3. Iron deficiency stimulates anthocyanin accumulation in grapevine apical leaves.

    Science.gov (United States)

    Caramanico, Leila; Rustioni, Laura; De Lorenzis, Gabriella

    2017-10-01

    Iron chlorosis is a diffuse disorder affecting Mediterranean vineyards. Beside the commonly described symptom of chlorophyll decrease, an apex reddening was recently observed. Secondary metabolites, such as anthocyanins, are often synthetized to cope with stresses in plants. The present work aimed to evaluate grapevine responses to iron deficiency, in terms of anthocyanin metabolism (reflectance spectrum, total anthocyanin content, HPLC profile and gene expression) in apical leaves of Cabernet sauvignon and Sangiovese grown in hydroponic conditions. Iron supply interruption produced after one month an increasing of anthocyanin content associated to a more stable profile in both cultivars. In Cabernet sauvignon, the higher red pigment accumulation was associated to a lower intensity of chlorotic symptoms, while in Sangiovese, despite the activation of the metabolism, the lower anthocyanin accumulation was associated to a stronger decrease in chlorophyll concentration. Gene expression data showed a significant increase of anthocyanin biosynthesis. The effects on the expression of structural and transcription factor genes of phenylpropanoid pathway were cultivar dependent. F3H, F3'H, F3'5'H and LDOX genes, in Cabernet sauvignon, and AOMT1 and AOMT genes, in Sangiovese, were positively affected by the treatment in response to iron deficiency. All data support the hypothesis of an anthocyanin biosynthesis stimulation rather than a decreased degradation of them due to iron chlorosis. Copyright © 2017 Elsevier Masson SAS. All rights reserved.

  4. Apical hypertrophic cardiomyopathy with hemodynamically unstable ventricular arrhythmia - Atypical presentation.

    Science.gov (United States)

    Chaturvedi, Hemant; Pandey, Rudra Dev; Sharma, Krishna Kumar; Makkar, Jitendra Singh; Sharma, Sanjeev K

    2016-09-01

    We present a patient with asymptomatic apical hypertrophic cardiomyopathy (AHCM) who recently developed cardiac arrhythmias, and shortly discuss the diagnostic modalities, differential diagnosis, and treatment strategy for this condition. AHCM is a rare form of hypertrophic cardiomyopathy, which usually involves the apex of the left ventricle. AHCM can occur with varied presentations such as chest pain, palpitations, dyspnea, syncope, atrial fibrillation, myocardial infarction, embolic events, ventricular fibrillation, and congestive heart failure. The most peculiar electrocardiogram findings are giant T-waves inversion in the precordial leads with left ventricular (LV) hypertrophy. A transthoracic echocardiogram is the initial diagnostic modality in the evaluation of AHCM and shows hypertrophy of the LV apex. Other diagnostic modalities, including left ventriculography, multislice spiral computed tomography, and cardiac magnetic resonance imagings, are also valuable tools. Medications used to manage include verapamil, beta-blockers, and antiarrhythmic agents. An implantable cardioverter defibrillator (ICD) is recommended for high-risk patients. Copyright © 2015 Cardiological Society of India. Published by Elsevier B.V. All rights reserved.

  5. Poda apical para uniformizar a colheita de flores de ?tango?

    Directory of Open Access Journals (Sweden)

    Francine Lorena Cuquel

    1999-01-01

    Full Text Available Este trabalho testou a poda apical das hastes de tango 4 e 6 semanas após a roçada, para homogeneizar a colheita que, comercialmente, necessita ser feita por um período de aproximadamente 5 dias numa mesma área. Foram avaliados altura média das plantas no primeiro dia de colheita, número médio de hastes colhidas/planta e a média do peso de matéria seca colhida/haste. Só foi verificada diferença significativa para altura média das plantas no primeiro dia de colheita, não se conseguindo reduzir o número de colheitas necessárias por planta. Existem indicações de que há necessidade de reduzir a variabilidade genética da população para homogeneizar a colheita de inflorescências de tango.

  6. Malignant Arrhythmia in Apical Ballooning Syndrome: Risk Factors and Outcomes

    Directory of Open Access Journals (Sweden)

    Samuel J. Asirvatham

    2008-08-01

    Full Text Available Objectives: We sought to determine the frequency and outcomes with symptomatic arrhythmia in patients with apical ballooning syndrome (ABS. Methods: A retrospective review of the Mayo Clinic Angiography database was conducted to identify patients who met the Mayo criteria for ABS. Patients with documented arrhythmias formed the study group, and 31 randomly selected patients with ABS but without arrhythmia formed the control group.Results: Out of 105 patients identified with ABS, 6 (5.7% women aged 69 +/- 9 years experienced significant arrhythmia (ventricular fibrillation, asystole, 2 patients died, and 1 required permanent pacemaker implantation. When compared with controls, the study group showed no significant difference with respect to ECG characteristics (QT, QRS duration or axis except for R-R interval variability (see comments below (30.6±6 vs 14.5±17 p = 0.0004, QTc, and P-R interval. Patients without arrhythmia were more likely to be on beta-blocker therapy than the study population (33% vs 80.6% p = 0.02. Conclusion: Life-threatening arrhythmia is uncommon (5.7% with ABS despite marked, structural abnormalities. When arrhythmias do occur, the outcome is poor. Prominent variability in R-R intervals appears to be predictive of significant arrhythmias in ABS. The role of beta-blocker therapy in preventing arrhythmia with ABS requires further investigation.

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

    International Nuclear Information System (INIS)

    Montero Aguilar, Mauricio; Zeledon Mayorga, Rodolfo; Ramirez Mora, Tatiana; Monestel Umana, Silvia

    2009-01-01

    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) [es

  8. Radiographic evaluation of apical root resorption following fixed orthodontic treatment

    Directory of Open Access Journals (Sweden)

    Sina Haghanifar

    2012-01-01

    Full Text Available Background and Aims: Apical root resorption is an adverse side effect of fixed orthodontic treatment which cannot be repaired. The aim of this study was to use panoramic radiographs to compare the root resorption before and after the orthodontic treatment with standard edgewise .018 appliance.Materials and Methods: The before and after treatment panoramic views of sixty-three patients needed fixed orthodontic treatment included 1520 teeth were categorized into 3 Grades (G0: without resorption, G1: mild resorption with blunt roots or ≤ 1/4 of root length, G2: moderate to severe resorption or > 1/4 to 1/2 of root length. Relationship between root resorption and sex and treatment duration was analyzed with Mann-whitney and Spearman's correlation coefficient, respectively.Results: The findings showed that 345 teeth were categorized as Grade 1. Grade 2 of root resorption was not found in this study. The highest amount of root resorption was recorded for the mandibular lateral incisor. In both gender, the root resorption of the mandible was more than that of the maxilla. The males showed significantly higher rate of resorption than the females (P0.05.Conclusion: The mandible and male patients showed higher amount of root resorption. In addition, root resorption was not related to the treatment duration and the side of the jaws.

  9. Embryology meets molecular biology: Deciphering the apical ectodermal ridge.

    Science.gov (United States)

    Verheyden, Jamie M; Sun, Xin

    2017-09-15

    More than sixty years ago, while studying feather tracks on the shoulder of the chick embryo, Dr. John Saunders used Nile Blue dye to stain the tissue. There, he noticed a darkly stained line of cells that neatly rims the tip of the growing limb bud. Rather than ignoring this observation, he followed it up by removing this tissue and found that it led to a striking truncation of the limb skeletons. This landmark experiment marks the serendipitous discovery of the apical ectodermal ridge (AER), the quintessential embryonic structure that drives the outgrowth of the limb. Dr. Saunders continued to lead the limb field for the next fifty years, not just through his own work, but also by inspiring the next generation of researchers through his infectious love of science. Together, he and those who followed ushered in the discovery of fibroblast growth factor (FGF) as the AER molecule. The seamless marriage of embryology and molecular biology that led to the decoding of the AER serves as a shining example of how discoveries are made for the rest of the developmental biology field. Copyright © 2017 Elsevier Inc. All rights reserved.

  10. Measurement of autoantibodies against human eye muscle plasma membranes in Graves' ophthalmopathy.

    OpenAIRE

    Faryna, M; Nauman, J; Gardas, A

    1985-01-01

    Antibodies that reacted with plasma membranes of human eye muscle but showed no binding to plasma membranes of human skeletal muscle were identified in serum of patients with Graves' ophthalmopathy. Rabbit antithyroglobulin serum at a dilution of 1 X 10(-3) to 20 X 10(-3) had no effect on the binding of these antibodies to eye muscle membrane antigens. There was no correlation between antihuman eye muscle plasma membrane antibodies and antihuman thyroid membrane antibodies or antibodies again...

  11. Immuno disc assay for screening duck hepatitis B surface antigen in serum, liver tissue and cultured hepatocytes

    NARCIS (Netherlands)

    G.A. de Wilde (G.); R.A. Heijtink

    1993-01-01

    textabstractAn immuno disc assay (IDA) for semi-quantitative analysis of the surface antigen (DHBsAg) of duck hepatitis B virus (DHBV) is described. Unpurified antigen preparations were adsorbed onto punched-out nitrocellulose membrane discs. Rabbit antiserum raised against serum-derived

  12. Prevalence of Weak D Antigen In Western Indian Population

    Directory of Open Access Journals (Sweden)

    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.

  13. 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

  14. Microfluidic squeezing for intracellular antigen loading in polyclonal B-cells as cellular vaccines

    Science.gov (United States)

    Lee Szeto, Gregory; van Egeren, Debra; Worku, Hermoon; Sharei, Armon; Alejandro, Brian; Park, Clara; Frew, Kirubel; Brefo, Mavis; Mao, Shirley; Heimann, Megan; Langer, Robert; Jensen, Klavs; Irvine, Darrell J.

    2015-05-01

    B-cells are promising candidate autologous antigen-presenting cells (APCs) to prime antigen-specific T-cells both in vitro and in vivo. However to date, a significant barrier to utilizing B-cells as APCs is their low capacity for non-specific antigen uptake compared to “professional” APCs such as dendritic cells. Here we utilize a microfluidic device that employs many parallel channels to pass single cells through narrow constrictions in high throughput. This microscale “cell squeezing” process creates transient pores in the plasma membrane, enabling intracellular delivery of whole proteins from the surrounding medium into B-cells via mechano-poration. We demonstrate that both resting and activated B-cells process and present antigens delivered via mechano-poration exclusively to antigen-specific CD8+T-cells, and not CD4+T-cells. Squeezed B-cells primed and expanded large numbers of effector CD8+T-cells in vitro that produced effector cytokines critical to cytolytic function, including granzyme B and interferon-γ. Finally, antigen-loaded B-cells were also able to prime antigen-specific CD8+T-cells in vivo when adoptively transferred into mice. Altogether, these data demonstrate crucial proof-of-concept for mechano-poration as an enabling technology for B-cell antigen loading, priming of antigen-specific CD8+T-cells, and decoupling of antigen uptake from B-cell activation.

  15. Primordial membranes

    DEFF Research Database (Denmark)

    Hanczyc, Martin M; Monnard, Pierre-Alain

    2017-01-01

    Cellular membranes, which are self-assembled bilayer structures mainly composed of lipids, proteins and conjugated polysaccharides, are the defining feature of cell physiology. It is likely that the complexity of contemporary cells was preceded by simpler chemical systems or protocells during...... the various evolutionary stages that led from inanimate to living matter. It is also likely that primitive membranes played a similar role in protocell 'physiology'. The composition of such ancestral membranes has been proposed as mixtures of single hydrocarbon chain amphiphiles, which are simpler versions...

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

    OpenAIRE

    P León; 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...

  17. Root lengthening with apical closure in two maxillary immature permanent central incisors after placement of mineral trioxide aggregate (MTA) as an apical plug.

    Science.gov (United States)

    Masmoudi, F; Bourmeche, I; Sebai, A; Baccouche, Z; Maatouk, F

    2018-02-01

    Necrosis of permanent immature teeth is a common reason for consultation in paediatric dentistry. Apexification is a therapeutic procedure aiming to create an apical calcified barrier in open apex teeth. Two cases are presented in which apexification were completed on immature permanent traumatised incisors using a mineral trioxide aggregate (MTA) plug. At 2 and 5 years showed apical closure and radicular elongation beyond the MTA plug. Unexpectedly, a regeneration of mineral tissues beyond the MTA plug occurred which is an uncommon outcome.

  18. The chicken erythrocyte-specific MHC antigen. Characterization and purification of the B-G antigen by monoclonal antibodies

    DEFF Research Database (Denmark)

    Salomonsen, J; Skjødt, K; Crone, M

    1987-01-01

    -labeled chicken erythrocyte membranes (CEM) followed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and autoradiography. The B-G antigen had an approximate molecular mass of 46-48 kd in reduced samples, depending on the haplotype, and in unreduced samples contained either dimers (85 kd......-G to be synthesized as a monomer, with dimerization taking place after 20-30 min. No change in the monomer's molecular mass due to posttranslational modifications was revealed. The antigen was purified from detergent extract of CEM by affinity chromatography with a monoclonal antibody, and then reduced and alkylated...... and affinity-purified once more. Finally, reverse-phase chromatography resulted in a pure product. The B-G antigen was identified in the various fractions by rocket immunoelectrophoresis. The final product was more than 99% pure, as estimated by SDS-PAGE analysis followed by silver stain of proteins. The yield...

  19. Risk Factors for Apical Periodontitis Sub-Urban Adult Population.

    Science.gov (United States)

    Oginni, A O; Adeleke, A A; Mejabi, M O; Sotunde, O A

    2015-06-01

    To assess the risk factors of apical periodontitis (AP) in a Nigerian sub-urban adult population and to compare the findings with those previously reported for various population groups. The study was based on a full mouth radiographic survey of 285 patients. Patients' age ranged from 18-60 years. All teeth were assessed individually and data recorded for caries, fractured / cracked teeth, root fillings, and tooth restorations. The gender, smoking habit, and frequency of dental visit were also recorded. Multiple logistic regression analyses were performed to identify predictors of AP in the individual. The prevalence of AP was 74.4%. The average number of teeth with AP per patient was 2.8 (range 1-5). AP was found to be more prevalent among people 40- 49 years old (87.2%). Primary carious lesions, fractured / cracked teeth, root fillings and coronal fillings were associated with the incidence of AP in the individual. Fractured teeth had a higher risk of developing AP than carious teeth. The presence of root fillings and coronal restorations were also associated with the development of AP. Smoking (OR=3.82; CI=2.17-6.75) and irregular dental visit (OR=6.73; CI=3.75-12.06) were statistically significant risk factors for developing AP. Gender was not a risk factor for AP (OR=0.86; CI=0.50-1.46). The prevalence of AP among adult Nigerians is slightly higher than reported figures for many Western societies. Fractured/cracked teeth had a higher risk of developing AP than carious teeth; hence patients with fractured / cracked teeth should seek treatment early to prevent the development of AP.

  20. Analysis of Microtubule Mediated Functions of Prostate Specific Membrane Antigen

    Science.gov (United States)

    2006-04-01

    supplemented with 10% fetal bovine serum, MEM nonessential amino acid solution (Invitrogen, Grand Island, NY), and penicillin/streptomycin. A7 and M2 cells were...results were obtained using immunoconjugates of other vaso - active agents, including recombinant interleukin-1h (IL-1h), physalaemin, histamine

  1. 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

    INTRODUCTION: Apical surgery is an important treatment option for teeth with post-treatment periodontitis. Although apical surgery involves root-end resection, no morphometric data are yet available about root-end resection and its impact on the root-to-crown ratio (RCR). The present study assessed...... 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....... The following parameters were assessed for all treated teeth as well as for specific tooth groups: length of root-end resection and percentage change of root length, preoperative and postoperative RCRs, and percentage change of RCR after apical surgery. RESULTS: The mean length of root-end resection was 3...

  2. Comparison of extracted teeth and simulated resin blocks on apical canal transportation.

    Science.gov (United States)

    Khalilak, Zohreh; Fallahdoost, Arjang; Dadresanfar, Bahareh; Rezvani, Gholamreza

    2008-01-01

    We aimed to compare apical canal transportation of extracted teeth and two types of simulated resin blocks. Mesiobuccal root of extracted maxillary molars, high hardness simulated resin blocks (Knoop hardness=40) and low hardness simulated resin blocks (Knoop hardness=22) were prepared with K-files using step-back technique (n=15 canals in each group). Double exposure radiographic technique was used for extracted teeth. Simulated resin blocks were stabilized and scanned before and after preparation. Pre and post-preparation pictures were superimposed and apical transportation was measured. The data were analyzed statistically using ANOVA and Tukey HSD tests. There was no significant difference in apical canal transportation between extracted teeth and high hardness resin blocks (P>0.05). Low hardness resin blocks showed more apical transportation than the other groups (Presin blocks were similar.

  3. Mineral trioxide aggregate-induced apical closure in nonvital immature permanent maxillary incisor

    Directory of Open Access Journals (Sweden)

    Meenu Bhola

    2017-01-01

    Full Text Available Treatment of nonvital immature permanent teeth with calcium hydroxide is associated with few difficulties such as weakened tooth root, root canal reinfection, and long treatment time. Mineral trioxide aggregate (MTA apical plug method is an alternative treatment method for open apices and has gained popularity in the recent times. This case report describes the management of a late-referral case of periapically involved, traumatized immature permanent incisor by endodontic treatment and the use of MTA apical plug. After preparing the access cavity, the working length was determined. The root canals were irrigated with 3% sodium hypochlorite and disinfected with metapex for 2 weeks. MTA was then placed in the apical 3 mm of the root canal. The remaining part of the root canal was filled with thermoplastic gutta-percha, and the coronal restoration was finished with composite resin. After 1-year follow-up, radiograph showed successful healing of periradicular radiolucency.

  4. Distribution and function of HCN channels in the apical dendritic tuft of neocortical pyramidal neurons.

    Science.gov (United States)

    Harnett, Mark T; Magee, Jeffrey C; Williams, Stephen R

    2015-01-21

    The apical tuft is the most remote area of the dendritic tree of neocortical pyramidal neurons. Despite its distal location, the apical dendritic tuft of layer 5 pyramidal neurons receives substantial excitatory synaptic drive and actively processes corticocortical input during behavior. The properties of the voltage-activated ion channels that regulate synaptic integration in tuft dendrites have, however, not been thoroughly investigated. Here, we use electrophysiological and optical approaches to examine the subcellular distribution and function of hyperpolarization-activated cyclic nucleotide-gated nonselective cation (HCN) channels in rat layer 5B pyramidal neurons. Outside-out patch recordings demonstrated that the amplitude and properties of ensemble HCN channel activity were uniform in patches excised from distal apical dendritic trunk and tuft sites. Simultaneous apical dendritic tuft and trunk whole-cell current-clamp recordings revealed that the pharmacological blockade of HCN channels decreased voltage compartmentalization and enhanced the generation and spread of apical dendritic tuft and trunk regenerative activity. Furthermore, multisite two-photon glutamate uncaging demonstrated that HCN channels control the amplitude and duration of synaptically evoked regenerative activity in the distal apical dendritic tuft. In contrast, at proximal apical dendritic trunk and somatic recording sites, the blockade of HCN channels decreased excitability. Dynamic-clamp experiments revealed that these compartment-specific actions of HCN channels were heavily influenced by the local and distributed impact of the high density of HCN channels in the distal apical dendritic arbor. The properties and subcellular distribution pattern of HCN channels are therefore tuned to regulate the interaction between integration compartments in layer 5B pyramidal neurons. Copyright © 2015 the authors 0270-6474/15/351024-14$15.00/0.

  5. Apical atrophy of retinal pigment epithelial detachments in central serous chorioretinopathy

    DEFF Research Database (Denmark)

    Pryds, Anders; Hamann, Steffen; Larsen, Michael

    2012-01-01

    compatible with CSC. RESULTS: A juxtafoveal retinal pigment epithelial detachment with apical atrophy of the retinal pigment epithelium with corresponding severe attenuation of the pigmentation of the retinal pigment epithelium was observed in both patients. One of the patients presented with a serous...... neurosensory retinal detachment with smokestack leakage 7 years after first being seen. The other patient was never seen with a neurosensory detachment. CONCLUSION: Isolated pigment epithelial detachment with apical retinal pigment epithelial atrophy may represent a precursor stage of CSC....

  6. Management of Teeth with Persistent Apical Periodontitis after Root Canal Treatment Using Regenerative Endodontic Therapy.

    Science.gov (United States)

    Saoud, Tarek Mohamed A; Huang, George T-J; Gibbs, Jennifer L; Sigurdsson, Asgeir; Lin, Louis M

    2015-10-01

    Regenerative endodontic therapy (RET) is currently used to treat immature teeth with necrotic pulp and/or apical periodontitis. However, recently RET has been used to treat mature teeth with necrotic pulp and/or apical periodontitis and resulted in regression of clinical signs and/or symptoms and resolution of apical periodontitis. The purpose of this case report was to describe the potential of using RET to treat 2 mature teeth with persistent apical periodontitis after root canal therapy using RET. Two male patients, one 26-year old and another 12-year old, presented for retreatment of persistent apical periodontitis after root canal treatment of 2 mature teeth (#9 and #19). The gutta-percha fillings in the canals of teeth #9 and #19 were removed with Carvene gutta-percha solvent (Prevest DenPro, Jammu, India) and ProTaper Universal rotary retreatment files (Dentsply Maillefer, Ballaigues, Switzerland). The canals of both teeth were further chemomechanically debrided with rotary retreatment files and copious amounts of sodium hypochlorite irrigation and dressed with Metapaste (Meta Biomed, Chungbuk, Korea). RET was performed on teeth #9 and #19. Periapical bleeding was provoked into the disinfected root canals. The blood clots were covered with mineral trioxide aggregate plugs, and the access cavities were restored with intermediate restorative material. Teeth #9 and #19 showed regression of clinical signs and/or symptoms and healing of apical periodontitis after 13-month and 14-month follow-ups, respectively. Tooth #9 revealed narrowing of the canal space and apical closure by deposition of hard tissue. RET has the potential to be used to retreat teeth with persistent apical periodontitis after root canal therapy. Copyright © 2015 American Association of Endodontists. Published by Elsevier Inc. All rights reserved.

  7. Pathogenicity of Exopolysaccharide-Producing Actinomyces oris Isolated from an Apical Abscess Lesion

    Science.gov (United States)

    2013-01-01

    testing and 16S ribosomal ribonucleic acid (rRNA) sequencing (Henssge et al. 2009). Materials and methods Bacteria A clinically isolated A. oris...described in many pathogens (Costerton et al. 1987). Although the bacteria in biofilms are considered to be dormant in general, it is conceivable that Actino...June 2012 Introduction Bacteria remaining in the apical part of the root canal are known to be involved in the development of apical abscesses (Nair

  8. Microscopic observations show invasion of inflammatory cells in the limb blastema and epidermis in pre-metamorphic frog tadpoles which destroy the Apical Epidermal CAP and impede regeneration.

    Science.gov (United States)

    Alibardi, Lorenzo

    2017-03-01

    Some limb regeneration in tadpoles of Rana dalmatina occurs at stages 44-48 when small hind-limbs are present while scarring occurs at stages 51-52 when forelimbs have developed and metamorphosis is approaching. Ultrastructural analysis of cells forming the regenerating blastema detects mesenchymal cells and an Apical Epidermal Cap (AEC) in regenerating limb blastema 5-6 days post-amputation at stages 46-48. In contrast, granulocytes and numerous macrophages and lymphocytes prevail over mesenchymal cells in limb blastema at stages 51-52, which are destined to form scars. An increase in inflammatory cells in limb blastema prior to metamorphosis suggests a negative influence of immune cells on limb regeneration. Inflammatory cells invade the apical wound epidermis where stem keratinocytes are likely destroyed, impeding the formation of an AEC, the microregion which leads to limb regeneration. The invasion of immune cells, however, may also represent a physiological consequence of the death of cell populations in the tadpoles occurring with approaching metamorphosis. The passage from an aquatic to a terrestrial life in this frog elicits the typical amniote scarring reaction after wounding, and the limb cannot regenerate. The present observations sustain the hypothesis that the evolution of the adaptive immunity in tetrapods while efficiently preserving adult self-condition, determined the loss of tissue regeneration since the embryonic antigens evocated in blastema cells are removed by immune cells of the adult. Copyright © 2016 Elsevier GmbH. All rights reserved.

  9. 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

  10. Abnormal apical-to-basal transport of dietary ovalbumin by secretory IgA stimulates a mucosal Th1 response.

    Science.gov (United States)

    Abed, J; Lebreton, C; Champier, G; Cuvillier, A; Cogné, M; Meresse, B; Dugave, C; Garfa-Traoré, M; Corthésy, B; Cerf-Bensussan, N; Heyman, M

    2014-03-01

    In celiac disease, enhanced permeability to gliadin peptides can result from their apico-basal transport by secretory immunoglobulin A1 (SIgA1) binding to the CD71 receptor ectopically expressed at the gut epithelial surface. Herein, we have established a mouse model in which there is apico-basal transport of the model antigen ovalbumin (OVA) by specific SIgA1 and have analyzed local T-cell activation. Transgenic DO11.10 mice were grafted with a hybridoma-secreting OVA-specific humanized IgA1, which could bind mouse CD71 and which were released in the intestinal lumen as SIgA. CD71 expression was induced at the gut apical surface by treating the mice with tyrphostin A8. Following gavage of the mice with OVA, OVA-specific CD4⁺ T cells isolated from the mesenteric lymph nodes displayed higher expression of the activation marker CD69 and produced more interferon gamma in mice bearing the hybridoma-secreting OVA-specific IgA1, than in ungrafted mice or in mice grafted with an irrelevant hybridoma. These results indicate that the protective role of SIgA1 might be jeopardized in human pathological conditions associated with ectopic expression of CD71 at the gut surface.

  11. 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...

  12. Effect of reciprocating systems and working lengths on apical microcrack development: a micro-CT Study

    Energy Technology Data Exchange (ETDEWEB)

    Oliveira, Bruna Paloma de; Câmara, Andréa Cruz; Duarte, Daniel Amancio; Antonino, Antonio Celso Dantas; Aguiar, Carlos Menezes, E-mail: bruna_paloma@msn.com [Universidade Federal de Pernambuco (UFPE), Recife, PE (Brazil); Heck, Richard John [Department of Land Resource Science, University of Guelph (Canada)

    2017-11-15

    The objective of this study was to evaluate the effect of root canal preparation with single-file reciprocating systems at different working lengths on the development of apical microcracks using micro-computed tomographic (micro-CT) imaging. Forty extracted human mandibular incisors were randomly assigned to 4 groups (n=10) according to the systems and working length used to prepare the root canals: Group A - WaveOne Gold at apical foramen (AF), Group B - WaveOne Gold 1 mm short of the AF (AF-1 mm), Group C - Unicone (AF) and Group D - Unicone (AF-1 mm). Micro-CT scanning was performed before and after root canal preparation at an isotropic resolution of 14 μm. Then, three examiners assessed the cross-sectional images generated to detect microcracks in the apical portion of the roots. Apical microcracks were visualized in 3, 1, 1, and 3 specimens in groups A, B, C, and D, respectively. All these microcracks observed after root canal preparation already existed prior to instrumentation, and no new apical microcrack was detected. For all groups, the number of slices presenting microcracks after root canal preparation was the same as before canal preparation. Root canal preparation with WaveOne Gold and Unicone, regardless of the working length, was not associated with apical microcrack formation. (author)

  13. Proliferative effects of apical, but not basal, matrix metalloproteinase-7 activity in polarized MDCK cells

    International Nuclear Information System (INIS)

    Harrell, Permila C.; McCawley, Lisa J.; Fingleton, Barbara; McIntyre, J. Oliver; Matrisian, Lynn M.

    2005-01-01

    Matrix metalloproteinase-7 (MMP-7) is primarily expressed in glandular epithelium. Therefore, its mechanism of action may be influenced by its regulated vectorial release to either the apical and/or basolateral compartments, where it would act on its various substrates. To gain a better understanding of where MMP-7 is released in polarized epithelium, we have analyzed its pattern of secretion in polarized MDCK cells expressing stably transfected human MMP-7 (MDCK-MMP-7), and HCA-7 and Caco2 human colon cancer cell lines. In all cell lines, latent MMP-7 was secreted to both cellular compartments, but was 1.5- to 3-fold more abundant in the basolateral compartment as compared to the apical. However, studies in the MDCK system demonstrated that MMP-7 activity was 2-fold greater in the apical compartment of MDCK-MMP-7 HIGH -polarized monolayers, which suggests the apical co-release of an MMP-7 activator. In functional assays, MMP-7 over-expression increased cell saturation density as a result of increased cell proliferation with no effect on apoptosis. Apical MMP-7 activity was shown to be responsible for the proliferative effect, which occurred, as demonstrated by media transfer experiments, through cleavage of an apical substrate and not through the generation of a soluble factor. Taken together, our findings demonstrate the importance of MMP-7 secretion in relation to its mechanism of action when expressed in a polarized epithelium

  14. Cancer antigen 125 and prognosis

    DEFF Research Database (Denmark)

    Høgdall, Estrid Vilma Solyom

    2008-01-01

    cancer antigen 125 determination may be implemented into clinical practice, cut-off levels must be evaluated and internationally defined. Studies examining serum cancer antigen 125 levels after surgery but before, during, or after treatment confirmed that changes in serum levels are of prognostic value...

  15. Radioimmunoassays of hidden viral antigens

    International Nuclear Information System (INIS)

    Neurath, A.R.; Strick, N.; Baker, L.; Krugman, S.

    1982-01-01

    Antigens corresponding to infectious agents may be present in biological specimens only in a cryptic form bound to antibodies and, thus, may elude detection. We describe a solid-phase technique for separation of antigens from antibodies. Immune complexes are precipitated from serum by polyethylene glycol, dissociated with NaSCN, and adsorbed onto nitrocellulose or polystyrene supports. Antigens remain topographically separated from antibodies after removal of NaSCN and can be detected with radiolabeled antibodies. Genomes from viruses immobilized on nitrocellulose can be identified by nucleic acid hybridization. Nanogram quantities of sequestered hepatitis B surface and core antigens and picogram amounts of hepatitis B virus DNA were detected. Antibody-bound adenovirus, herpesvirus, and measles virus antigens were discerned by the procedure

  16. Four Cation-Selective Transporters Contribute to Apical Uptake and Accumulation of Metformin in Caco-2 Cell Monolayers

    Science.gov (United States)

    Han, Tianxiang (Kevin); Proctor, William R.; Costales, Chester L.; Cai, Hao; Everett, Ruth S.

    2015-01-01

    Metformin is the frontline therapy for type II diabetes mellitus. The oral bioavailability of metformin is unexpectedly high, between 40 and 60%, given its hydrophilicity and positive charge at all physiologic pH values. Previous studies in Caco-2 cell monolayers, a cellular model of the human intestinal epithelium, showed that during absorptive transport metformin is taken up into the cells via transporters in the apical (AP) membrane; however, predominant transport to the basolateral (BL) side occurs via the paracellular route because intracellular metformin cannot egress across the BL membrane. Furthermore, these studies have suggested that the AP transporters can contribute to intestinal accumulation and absorption of metformin. Transporter-specific inhibitors as well as a novel approach involving a cocktail of transporter inhibitors with overlapping selectivity were used to identify the AP transporters that mediate metformin uptake in Caco-2 cell monolayers; furthermore, the relative contributions of these transporters in metformin AP uptake were also determined. The organic cation transporter 1, plasma membrane monoamine transporter (PMAT), serotonin reuptake transporter, and choline high-affinity transporter contributed to approximately 25%, 20%, 20%, and 15%, respectively, of the AP uptake of metformin. PMAT-knockdown Caco-2 cells were constructed to confirm the contribution of PMAT in metformin AP uptake because a PMAT-selective inhibitor is not available. The identification of four intestinal transporters that contribute to AP uptake and potentially intestinal absorption of metformin is a significant novel finding that can influence our understanding of metformin pharmacology and intestinal drug-drug interactions involving this highly prescribed drug. PMID:25563903

  17. LOCALIZATION OF ANTIGEN IN TISSUE CELLS

    Science.gov (United States)

    Coons, Albert H.; Leduc, Elizabeth H.; Kaplan, Melvin H.

    1951-01-01

    The fate of three proteins, crystalline hen's egg albumin, crystalline bovine plasma albumin, and human plasma γ-globulin, was traced after intravenous injection into mice. This was done by preparing frozen sections of quick-frozen tissue, allowing what foreign protein might be present in the section to react with homologous antibody labelled with fluorescein, and examining the section under the fluorescence microscope. By this means, which employs the serological specificity of the protein as a natural "marker," all three of these proteins were found in the cells of the reticulo-endothelial system, the connective tissue, the vascular endothelium, the lymphocytes of spleen and lymph node, and the epithelium of the kidney tubules, the liver, and in very small amounts in the adrenal. The central nervous system was not studied. All three persisted longest in the reticulo-endothelial system and the connective tissue, and in the doses employed egg white (10 mg.) was no longer detectable after 1 day, bovine albumin (10 mg.) after 2 days, and human γ-globulin (4 mg.) after 6 days, although in a somewhat higher dose (10 mg.) human γ-globulin persisted longer than 8 days. Egg albumin differed from the others in not being detectable in the cells of the renal glomerulus. It was found that each of the three proteins was present in the nuclei of each cell type enumerated above, often in higher concentration than in the cytoplasm. Further, some of the nuclei not only contained antigen, soon after injection, but were also surrounded by a bright ring associated with the nuclear membrane. By means of photographic records under the fluorescence microscope of sections stained for antigen, and direct observation under the light microscope of the same field subsequently stained with hematoxylin and eosin, it could be determined that the antigen was not adsorbed to chromatin or nucleoli, but was apparently in solution in the nuclear sap. PMID:14803641

  18. Chemical camouflage of antigenic determinants: stealth erythrocytes.

    Science.gov (United States)

    Scott, M D; Murad, K L; Koumpouras, F; Talbot, M; Eaton, J W

    1997-07-08

    In a number of clinical circumstances it would be desirable to artificially conceal cellular antigenic determinants to permit survival of heterologous donor cells. A case in point is the problem encountered in transfusions of patients with rare blood types or chronically transfused patients who become allosensitized to minor blood group determinants. We have tested the possibility that chemical modification of the red blood cell (RBC) membrane might serve to occlude antigenic determinants, thereby minimizing transfusion reactions. To this end, we have covalently bound methoxy(polyethylene glycol) (mPEG) to the surface of mammalian RBC via cyanuric chloride coupling. Human RBC treated with this technique lose ABO blood group reactivity as assessed by solution-phase antisera agglutination. In accord with this, we also find a profound decrease in anti-blood group antibody binding. Furthermore, whereas human monocytes avidly phagocytose untreated sheep RBC, mPEG-derivatized sheep RBC are ineffectively phagocytosed. Surprisingly, human and mouse RBC appear unaffected by this covalent modification of the cell membrane. Thus, mPEG-treated RBC are morphologically normal, have normal osmotic fragility, and mPEG-derivatized murine RBC have normal in vivo survival, even following repeated infusions. Finally, in preliminary experiments, mPEG-modified sheep RBC intraperitoneally transfused into mice show significantly improved (up to 360-fold) survival when compared with untreated sheep RBC. We speculate that similar chemical camouflage of intact cells may have significant clinical applications in both transfusion (e.g., allosensitization and autoimmune hemolytic disease) and transplantation (e.g., endothelial cells and pancreatic beta cells) medicine.

  19. Mature IgM-expressing plasma cells sense antigen and develop competence for cytokine production upon antigenic challenge

    Science.gov (United States)

    Blanc, Pascal; Moro-Sibilot, Ludovic; Barthly, Lucas; Jagot, Ferdinand; This, Sébastien; de Bernard, Simon; Buffat, Laurent; Dussurgey, Sébastien; Colisson, Renaud; Hobeika, Elias; Fest, Thierry; Taillardet, Morgan; Thaunat, Olivier; Sicard, Antoine; Mondière, Paul; Genestier, Laurent; Nutt, Stephen L.; Defrance, Thierry

    2016-01-01

    Dogma holds that plasma cells, as opposed to B cells, cannot bind antigen because they have switched from expression of membrane-bound immunoglobulins (Ig) that constitute the B-cell receptor (BCR) to production of the secreted form of immunoglobulins. Here we compare the phenotypical and functional attributes of plasma cells generated by the T-cell-dependent and T-cell-independent forms of the hapten NP. We show that the nature of the secreted Ig isotype, rather than the chemical structure of the immunizing antigen, defines two functionally distinct populations of plasma cells. Fully mature IgM-expressing plasma cells resident in the bone marrow retain expression of a functional BCR, whereas their IgG+ counterparts do not. Antigen boost modifies the gene expression profile of IgM+ plasma cells and initiates a cytokine production program, characterized by upregulation of CCL5 and IL-10. Our results demonstrate that IgM-expressing plasma cells can sense antigen and acquire competence for cytokine production upon antigenic challenge. PMID:27924814

  20. Polyclonal Antibody Production for Membrane Proteins via Genetic Immunization.

    Science.gov (United States)

    Hansen, Debra T; Robida, Mark D; Craciunescu, Felicia M; Loskutov, Andrey V; Dörner, Katerina; Rodenberry, John-Charles; Wang, Xiao; Olson, Tien L; Patel, Hetal; Fromme, Petra; Sykes, Kathryn F

    2016-02-24

    Antibodies are essential for structural determinations and functional studies of membrane proteins, but antibody generation is limited by the availability of properly-folded and purified antigen. We describe the first application of genetic immunization to a structurally diverse set of membrane proteins to show that immunization of mice with DNA alone produced antibodies against 71% (n = 17) of the bacterial and viral targets. Antibody production correlated with prior reports of target immunogenicity in host organisms, underscoring the efficiency of this DNA-gold micronanoplex approach. To generate each antigen for antibody characterization, we also developed a simple in vitro membrane protein expression and capture method. Antibody specificity was demonstrated upon identifying, for the first time, membrane-directed heterologous expression of the native sequences of the FopA and FTT1525 virulence determinants from the select agent Francisella tularensis SCHU S4. These approaches will accelerate future structural and functional investigations of therapeutically-relevant membrane proteins.

  1. NovelTreponema pallidumRecombinant Antigens for Syphilis Diagnostics: Current Status and Future Prospects.

    Science.gov (United States)

    Kubanov, Aleksey; Runina, Anastassia; Deryabin, Dmitry

    2017-01-01

    The recombinant protein technology considerably promoted the development of rapid and accurate treponema-specific laboratory diagnostics of syphilis infection. For the last ten years, the immunodominant recombinant inner membrane lipoproteins are proved to be sensitive and specific antigens for syphilis screening. However, the development of an enlarged T. pallidum antigen panel for diagnostics of early and late syphilis and differentiation of syphilis stages or cured syphilis remains as actual goal of multidisciplinary expertise. Current review revealed novel recombinant antigens: surface-exposed proteins, adhesins, and periplasmic and flagellar proteins, which are promising candidates for the improved syphilis serological diagnostics. The opportunities and limitations of diagnostic usage of these antigens are discussed and the criteria for selection of optimal antigens panel summarized.

  2. Novel Treponema pallidum Recombinant Antigens for Syphilis Diagnostics: Current Status and Future Prospects

    Science.gov (United States)

    Kubanov, Aleksey; Runina, Anastassia

    2017-01-01

    The recombinant protein technology considerably promoted the development of rapid and accurate treponema-specific laboratory diagnostics of syphilis infection. For the last ten years, the immunodominant recombinant inner membrane lipoproteins are proved to be sensitive and specific antigens for syphilis screening. However, the development of an enlarged T. pallidum antigen panel for diagnostics of early and late syphilis and differentiation of syphilis stages or cured syphilis remains as actual goal of multidisciplinary expertise. Current review revealed novel recombinant antigens: surface-exposed proteins, adhesins, and periplasmic and flagellar proteins, which are promising candidates for the improved syphilis serological diagnostics. The opportunities and limitations of diagnostic usage of these antigens are discussed and the criteria for selection of optimal antigens panel summarized. PMID:28523273

  3. Monoclonal antibodies to antigens on human neutrophils, activated T lymphocytes, and acute leukemia blast cells

    International Nuclear Information System (INIS)

    Miterev, G.Yu.; Burova, G.F.; Puzhitskaya, M.S.; Danilevich, S.V.; Bulycheva, T.I.

    1987-01-01

    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 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

  4. 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.

  5. 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.

  6. Colonoscopy and carcinoembryonic antigen variations.

    Science.gov (United States)

    Sousa, Rita G; Nunes, Ana; Meira, Tânia; Carreira, Olga; Pires, Ana M; Freitas, João

    2014-01-01

    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. To evaluate the influence of bowel preparation and colonoscopy in carcinoembryonic antigen blood levels. 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. 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). 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.

  7. Apical foramen morphology according to the length of merged canal at the apex

    Directory of Open Access Journals (Sweden)

    Hee-Ho Kim

    2013-02-01

    Full Text Available Objectives The aim of this study was to investigate the relationship between the apical foramen morphology and the length of merged canal at the apex in type II root canal system. Materials and Methods This study included intact extracted maxillary and mandibular human premolars (n = 20 with fully formed roots without any visible signs of external resorption. The root segments were obtained by removing the crown 1 mm beneath the cementum-enamel junction (CEJ using a rotary diamond disk. The distance between the file tip and merged point of joining two canals was defined as Lj. The roots were carefully sectioned at 1 mm from the apex by a slow-speed water-cooled diamond saw. All cross sections were examined under the microscope at ×50 magnification and photographed to estimate the shape of the apical foramen. The longest and the shortest diameter of apical foramen was measured using ImageJ program (1.44p, National Institutes of Health. Correlation coefficient was calculated to identify the link between Lj and the apical foramen shape by Pearson's correlation. Results The average value of Lj was 3.74 mm. The average of proportion (P, estimated by dividing the longest diameter into the shortest diameter of the apical foramen, was 3.64. This study showed a significant negative correlation between P and Lj (p < 0.05. Conclusions As Lj gets longer, the apical foramen becomes more ovally shaped. Likewise, as it gets shorter, the apical foramen becomes more flat shaped.

  8. Clinical significance of left ventricular apical aneurysms in hypertrophic cardiomyopathy patients: the role of diagnostic electrocardiography.

    Science.gov (United States)

    Ichida, Masaru; Nishimura, Yoshioki; Kario, Kazuomi

    2014-10-01

    Some patients with hypertrophic cardiomyopathy (HCM) develop left ventricular apical aneurysm, leading to serious cardiovascular complications. The aims of this study were to identify the incidence and clinical course of HCM patients with apical aneurysms in Japan, and to evaluate the role of electrocardiography (ECG) as a screening test to detect apical aneurysms in HCM patients. In a retrospective, single center analysis of a population of 247 HCM patients, 21 patients (8.5%) had left ventricular apical aneurysms. Their mean age was 60 ± 14 years (range: 23-77 years) at study entry. Over 4.7 ± 3.3 years of follow-up, 10 patients (47.6%) experienced an adverse clinical event (annual event rate: 10.1%/y), including five implantable cardioverter-defibrillator (ICD) implantations for ventricular tachycardia/ventricular fibrillation (VT/VF), an appropriate discharge of ICD for VT/VF, and four nonfatal thromboembolic strokes. Two patients developed systolic dysfunction (ejection fraction <50%). No sudden cardiac death or progressive heart failure was detected. Fourteen patients showed ST-segment elevation (≥ 1 mm) in V3 through V5 of ECG. In four patients, progression of the ST-segment elevation was recognized. When the ST-segment elevation was used to identify apical aneurysms in HCM patients, the sensitivity was 66.7%, and the specificity was 98.7%. Apical aneurysms in HCM patients in Japan are not rare, and are associated with serious cardiovascular complications. The early diagnosis of apical aneurysms can be achieved by serial ECG. Copyright © 2014 Japanese College of Cardiology. Published by Elsevier Ltd. All rights reserved.

  9. 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.

  10. Size and geometry of apical sesamoid fracture fragments as a determinant of prognosis in Thoroughbred racehorses.

    Science.gov (United States)

    Kamm, J L; Bramlage, L R; Schnabel, L V; Ruggles, A J; Embertson, R M; Hopper, S A

    2011-07-01

    Analysis was performed to examine a method for refining the preoperative prognosis for horses that had surgery to remove apical fractures of the proximal sesamoid bones (PSBs). To determine if: 1) there was a difference in size or configuration of apical fractures between the different anatomical locations of the PSBs, which have been shown to affect the prognosis; and 2) the size or configuration could predict the prognosis for racehorses with these fractures. The study included 110 weanlings and yearlings and 56 training racehorses that underwent surgery to remove apical PSB fractures. Radiographs of the fractures were used for measurement of the abaxial and axial proportion and the abaxial to axial ratio, and race records were used to determine average earnings per start (AEPS) and total post operative starts. Analysis of variance and regression statistics were used to compare the fragment sizes between the specific PSBs on each of the limbs and compare size and configuration of the fractures to prognosis. There was a significantly larger abaxial to axial ratio (more transverse fracture) for the forelimb medial sesamoids than for all other sesamoids in untrained racehorses (P = 0.03). There were no other significant differences in size. There was no relationship between fracture size or configuration and AEPS nor total post operative starts. Apical fractures in weanlings and yearlings tend to be more transverse in the forelimb medial PSBs than the other PSBs. Apical fracture size and geometry does not determine prognosis for apical sesamoid fractures. Horses that undergo surgery to remove larger apical fractures of the PSBs do not have a worse outcome than those horses with smaller fractures. © 2010 EVJ Ltd.

  11. Antimicrobial photodynamic therapy for the treatment of teeth with apical periodontitis: a histopathological evaluation.

    Science.gov (United States)

    Silva, Lea Assed Bezerra; Novaes, Arthur B; de Oliveira, Rafael R; Nelson-Filho, Paulo; Santamaria, Milton; Silva, Raquel Assed Bezerra

    2012-03-01

    This study evaluated the in vivo response of apical and periapical tissues of dogs' teeth with apical periodontitis after one-session endodontic treatment with and without antimicrobial photodynamic therapy (aPDT). Sixty root canals with experimentally induced apical periodontitis were instrumented and assigned to 4 groups receiving aPDT and root canal filling (RCF) or not: group aPDT+/RCF+ (n = 20): aPDT (photosensitizer phenothiazine chloride at 10 mg/mL for 3 minutes and diode laser [λ = 660 nm, 60 mW/cm(2)] for 1 minute) and RCF in the same session; group aPDT+/RCF- (n = 10); group aPDT-/RCF+ (n = 20), and group aPDT-/RCF- (n = 10). Teeth were restored, and the animals were killed after 90 days. Sections from the maxillas and mandibles were stained with hematoxylin-eosin and Mallory trichrome and examined under light microscopy. Descriptive (ie, newly formed apical mineralized tissue, periapical inflammatory infiltrate, apical periodontal ligament thickness, and mineralized tissue resorption) and quantitative (ie, periapical lesion size and number of inflammatory cells) microscopic analysis was performed. Quantitative data were analyzed by the Kruskal-Wallis and Dunn tests (α = .05). In the aPDT-treated groups, the periapical region was moderately/severely enlarged with no inflammatory cells, moderate neoangiogenesis and fibrogenesis, and the smallest periapical lesions. Although apical closure by mineralized tissue deposition was not achieved, the absence of inflammatory cells, moderate neoangiogenesis, and fibrogenesis in the periapical region in the groups treated with aPDT indicate that this can be a promising adjunct therapy to cleaning and shaping procedures in teeth with apical periodontitis undergoing one-session endodontic treatment. Copyright © 2012 American Association of Endodontists. Published by Elsevier Inc. All rights reserved.

  12. 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.

  13. 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...

  14. Targeted disruption of core 1 β1,3-galactosyltransferase (C1galt1 induces apical endocytic trafficking in human corneal keratinocytes.

    Directory of Open Access Journals (Sweden)

    Ana Guzman-Aranguez

    Full Text Available BACKGROUND: Exposed mucosal surfaces limit constitutive endocytosis under physiological conditions to prevent uptake of macromolecules and pathogens and, therefore, cellular damage. It is now accepted that cell surface mucins, a group of high molecular weight glycoproteins on the epithelial glycocalyx, defined by their extensive O-glycosylation, play a major role in maintaining barrier function in these surfaces, but the precise mechanisms are unclear. METHODOLOGY/PRINCIPAL FINDINGS: In this work, we utilized a stable tetracycline-inducible RNA interfering system targeting the core 1 ß1,3-galactosyltransferase (C1galt1 or T-synthase, a critical galactosyltransferase required for the synthesis of core 1 O-glycans, to explore the role of mucin-type carbohydrates in apical endocytic trafficking in human corneal keratinocytes. Using cell surface biotinylation and subcellular fractionation, we found increased accumulation of plasma membrane protein in endosomes after C1galt1 depletion. Confocal laser scanning microscopy and fluorometry revealed increased translocation of negatively charged fluorescent nanospheres after C1galt1 knockdown sustained by an active transport process and largely independent of apical intercellular junctions. Internalization of nanospheres could be blocked by dynasore, nocodazole, chlorpromazine, and hyperosmotic sucrose, suggesting a mechanism for clathrin-coated pit budding and vesicular trafficking. This possibility was supported by experiments showing nanosphere colocalization with clathrin heavy chain in the cytoplasm. CONCLUSIONS/SIGNIFICANCE: Together, the data suggest that core 1 O-glycans contribute to maintenance of apical barrier function on exposed mucosal surfaces by preventing clathrin-mediated endocytosis.

  15. 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.

  16. En face optical coherence tomography investigation of apical microleakage after laser-assisted endodontic treatment.

    Science.gov (United States)

    Todea, Carmen; Balabuc, Cosmin; Sinescu, Cosmin; Filip, Laura; Kerezsi, Cristina; Calniceanu, Mircea; Negrutiu, Meda; Bradu, Adrian; Hughes, Michael; Podoleanu, Adrian Gh

    2010-09-01

    The aim of our study was to evaluate the potential of en face optical coherence tomography (OCT) for the detection of apical microleakage after 980 nm and 1,064 nm laser-assisted endodontic treatment. Ninety, human, single-rooted teeth with one straight root canal and closed apices were used. All roots were prepared biomechanically to the working length at an apical size 30 and 0.06 taper. The teeth were divided into three equal groups of 30 samples each, according to the treatment to be applied to the root canal. Group I received 980 nm diode laser (3 W, 0.01 s on time, 0.01 s off time, 5 s per procedure, four procedures); group II received neodymium:yttrium-aluminum-garnet (Nd:YAG) laser (1.5 W, 15 Hz, 5 s per procedure, four procedures). In group III the root canals were approached conventionally only. In all groups the root canal filling was performed with AH Plus endodontic sealer and gutta-percha points. An en face OCT prototype was used for the investigation of apical microleakage. According to one-way analysis of variance (ANOVA) and en face OCT, the number of defects in the laser groups was significantly lower (P laser groups (P = 0.049). En face OCT imaging proved that laser-assisted endodontic treatment improved the prognosis of root canal filling and led to a reduction in apical microleakage.

  17. Apical Dimension of Root Canal Clinically Assessed with and without Periapical Lesions

    Directory of Open Access Journals (Sweden)

    Andrea Gesi

    2014-01-01

    Full Text Available To clinically evaluate the dimension of the more apical extent of the root canal after appropriate preflaring in the case of primary treatment and retreatment with and without the presence of periapical radiolucency, 392 single-rooted teeth with only one canal were evaluated during endodontic therapy. The canals were divided in two groups depending on the presence or absence of periapical radiolucency. After preflaring of the root canal the size of the root canal terminus (apical canal dimension was gauged with hand-held Light Speed LS1 files inserted at the estimated working length and established with the use of an electronic apex locator. The dimension recorded in the computer database was represented by the largest instrument able to reach the electronically established working length. The differences between the treatment groups were assessed using the Mann-Whitney U test and the significance level was set at P<0.05. Teeth with lesions had a significantly greater diameter in the apical region than teeth without lesions (P<0.001. The dimension of the apical portion of the root canal is larger in the case of periapical radiolucency. This involves verifying this parameter in order to use the correct sized instruments and to obtain an efficient cutting action at the apical level.

  18. Effects of apical meristem loss on sylleptic branching and growth of hybrid poplar

    Energy Technology Data Exchange (ETDEWEB)

    Zeleznik, Joseph D. [North Dakota State University, Fargo, ND (United States). Plant Sciences Department

    2007-07-15

    The effects of apical meristem loss on the growth and development of hybrid poplar trees was investigated. This was done by clipping back either the apical meristem alone (dividing cells), or the apical meristem plus a small amount of additional stem tissue (expanding cells, <1 cm), at various times during the first growing season. Two clones (NM6-nonsylleptic habit, and DN34-slightly sylleptic habit) were tested at close spacing (0.6 m) in the nursery. Clipping generally increased the number of sylleptic branches formed. Clipping 69 days after planting resulted in the largest number of sylleptic branches while clipping 4 weeks later gave no increase in syllepsis. Clipping temporarily reduced height growth of both clones but total height at the end of the first growing season was not affected by any treatment. There were some slight differences in growth during the second growing season; despite these differences, total stem biomass and total tree biomass after 2 years were not affected by temporary loss of the apical meristem in the first growing season. Results suggest that death or removal of hybrid poplar apical meristems by tip borers or ungulates has no long-term effects on aboveground growth as measured by height or biomass. (author)

  19. Effects of Different Rotary Files Combined with Different Irrigation Needles on Apically Extruded Debris.

    Science.gov (United States)

    Uzunoglu, Emel; Turker, Sevinc Aktemur; Görduysus, Melahat

    2015-01-01

    The aim of this study was to compare the apically extruded debris associated with ProTaper Next and OneShape Apical files using two different irrigation needles, open-ended and closed-ended (Max-I-Probe). Forty-eight mandibular premolars were assigned to four groups (n=12) as follows: Group PTN-SN: Root canal was prepared with ProTaper Next (PTN) and irrigated with open-ended standard needle; Group PTN-MP: Root canal was prepared with PTN and irrigated with Max-I-Probe; Group OSA-SN: Root canal was prepared with OneShape Apical (OSA) and irrigated with open-ended needle; Group OSA-MP: Root canal was prepared with OSA and irrigated with Max-I-Probe. Debris extruded during instrumentation was collected into pre-weighed Eppendorf tubes. After storage in an incubator at 70 °C for 5 days, the Eppendorf tubes were weighed to obtain the final weight with extruded debris. The difference between pre-and post-debris weights was calculated and statistical analysis was performed using Kruskal-Wallis and Mann-Whitney U tests (pextruded debris (p>0.05). OneShape Apical and ProTaper Next resulted in extrusion of debris beyond the apical foramen, regardless of needle tip.

  20. Apical deficiency triggers JNK-dependent apoptosis in the embryonic epidermis of Drosophila

    Science.gov (United States)

    Kolahgar, Golnar; Bardet, Pierre-Luc; Langton, Paul F.; Alexandre, Cyrille; Vincent, Jean-Paul

    2011-01-01

    Epithelial homeostasis and the avoidance of diseases such as cancer require the elimination of defective cells by apoptosis. Here, we investigate how loss of apical determinants triggers apoptosis in the embryonic epidermis of Drosophila. Transcriptional profiling and in situ hybridisation show that JNK signalling is upregulated in mutants lacking Crumbs or other apical determinants. This leads to transcriptional activation of the pro-apoptotic gene reaper and to apoptosis. Suppression of JNK signalling by overexpression of Puckered, a feedback inhibitor of the pathway, prevents reaper upregulation and apoptosis. Moreover, removal of endogenous Puckered leads to ectopic reaper expression. Importantly, disruption of the basolateral domain in the embryonic epidermis does not trigger JNK signalling or apoptosis. We suggest that apical, not basolateral, integrity could be intrinsically required for the survival of epithelial cells. In apically deficient embryos, JNK signalling is activated throughout the epidermis. Yet, in the dorsal region, reaper expression is not activated and cells survive. One characteristic of these surviving cells is that they retain discernible adherens junctions despite the apical deficit. We suggest that junctional integrity could restrain the pro-apoptotic influence of JNK signalling. PMID:21693518

  1. Antigenic relationships among four herpesviruses.

    Science.gov (United States)

    Blue, W T; Plummer, G

    1973-06-01

    Common viral antigens were detected, by fluorescent-antibody studies, in cells infected with herpes simplex virus 1, squirrel monkey herpesvirus 1, bovine rhinotracheitis, and equine abortion viruses. The two primate viruses showed slight cross-neutralization.

  2. HLA-B27 antigen

    Science.gov (United States)

    Human leukocyte antigen B27; Ankylosing spondylitis-HLA; Psoriatic arthritis-HLA; Reactive arthritis-HLA ... Erythrocyte sedimentation rate ( ESR ) Rheumatoid factor X-rays HLA testing is also used to match donated tissue ...

  3. How far is the root apex of a unilateral impacted canine from the root apices' arch form?

    Science.gov (United States)

    Kim, Sung-Hun; Kim, You-Min; Oh, Sewoong; Kim, Seong-Sik; Park, Soo-Byung; Son, Woo-Sung; Kim, Yong-Il

    2017-02-01

    The purpose of this study was to determine the arch form of the root apices of normally erupting teeth and then determine the differences in the location of the apex of impacted canines relative to normally erupting canines. In addition, we sought to determine whether the labiopalatal position of the impacted canines influences the position of the apices. The study included 21 patients with unerupted canines that subsequently had a normal eruption, 21 patients with palatally impacted canines, 27 patients with labially impacted canines, and 17 patients with midalveolus impacted canines. Images were obtained using cone beam computed tomography, and the x, y, and z coordinates of the root apices were determined using Ondemand3D software (Cybermed Co., Seoul, Korea). Two-dimensional coordinates were converted from acquired 3-dimensional coordinates via projection on a palatal plane, and the Procrustes method was used to process the converted 2-dimensional coordinates and to draw the arch forms of the root apices. Finally, we measured the extent of root apex deviation from the arch forms of the root apices. Normally erupting canines showed that even though calcifications may be immature, their positions were aligned with a normal arch form. The root apices of the impacted canines were an average of 6.572 mm away from the root apices' arch form, whereas those of the contralateral nonimpacted canines were an average distance of 2.221 mm away, a statistically significant difference. The palatally impacted canines' root apices distribution tended toward the first premolar root apices. Incompletely calcified, unerupted teeth with a subsequent normal eruption showed a normal arch form of the root apices. The root apices of impacted canines were farther from the arch forms than were the nonimpacted canines. Also, the root apices of impacted canines in the palatal area showed distributions different from those of the other impacted canine groups. Copyright © 2017 American

  4. 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.

  5. Natural selection promotes antigenic evolvability.

    Science.gov (United States)

    Graves, Christopher J; Ros, Vera I D; Stevenson, Brian; Sniegowski, Paul D; Brisson, Dustin

    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 an experimentally tractable system to test whether natural selection has favored mechanisms that increase evolvability. Many antigenic variation systems consist of paralogous unexpressed 'cassettes' that recombine into an expression site to rapidly alter the expressed protein. Importantly, the magnitude of antigenic change is a function of the genetic diversity among the unexpressed cassettes. Thus, evidence that selection favors among-cassette diversity is direct evidence that natural selection promotes antigenic evolvability. We used the Lyme disease bacterium, Borrelia burgdorferi, as a model to test the prediction that natural selection favors amino acid diversity among unexpressed vls cassettes and thereby promotes evolvability in a primary surface antigen, VlsE. The hypothesis that diversity among vls cassettes is favored by natural selection was supported in each B. burgdorferi strain analyzed using both classical (dN/dS ratios) and Bayesian population genetic analyses of genetic sequence data. This hypothesis was also supported by the conservation of highly mutable tandem-repeat structures across B. burgdorferi strains despite a near complete absence of sequence conservation. Diversification among vls cassettes due to natural selection and mutable repeat structures promotes long-term antigenic evolvability of VlsE. These findings provide a direct demonstration that molecular mechanisms that enhance evolvability of surface antigens are an evolutionary adaptation. The molecular evolutionary processes identified here can serve as a model for the evolution of antigenic evolvability in many pathogens which utilize similar strategies to establish chronic infections.

  6. Epicutaneous sensitization with protein antigen

    Directory of Open Access Journals (Sweden)

    I-Lin Liu

    2012-12-01

    Full Text Available In the past few decades there has been a progressive understanding that epicutaneous sensitization with protein antigen is an important sensitization route in patients with atopic dermatitis. A murine protein-patch model has been established, and an abundance of data has been obtained from experiments using this model. This review discusses the characteristics of epicutaneous sensitization with protein antigen, the induced immune responses, the underlying mechanisms, and the therapeutic potential.

  7. Apical Microleakage in Root Canals Obturated With 2 Different Endodontic Sealer Systems in Canine Teeth of Dogs.

    Science.gov (United States)

    Lothamer, Chad W; Anderson, Ashley; Hetzel, Scott J; Adrian, Alex I; Snyder, Christopher; Goldschmidt, Stephanie; Soukup, Jason W

    2017-06-01

    An apical dye leakage test was utilized to compare 2 endodontic sealer systems commonly used in veterinary endodontic treatment. Rotary instrumentation followed by injection of 2 different sealers (GuttaFlow 2 and AH Plus) and a single master cone obturation technique were compared. Following obturation and restoration, specimens were immersed in India ink for 48 hours. A tooth mineral clearing technique was utilized to render the tooth samples transparent. Subsequently, the apices were evaluated for prevalence and magnitude of apical dye penetration under magnification. Statistical analysis of the results revealed no significant differences in the prevalence or magnitude of apical dye penetration between groups.

  8. Bacillus thuringiensis Cry1A toxin-binding glycoconjugates present on the brush border membrane and in the peritrophic membrane of the Douglas-fir tussock moth are peritrophins

    Science.gov (United States)

    Algimantas P. Valaitis; John D. Podgwaite

    2013-01-01

    Bacillus thuringiensis (Bt) Cry1A toxin-binding sites in the Douglas fir tussock moth (DFTM) larval gut were localized using immunofluorescence microscopy. Cry1Aa, Cry1Ab and Cry1Ac all bound strongly to the DFTM peritrophic membrane (PM); weaker binding of the Cry1A toxins was observed along the apical brush border of the midgut epithelium....

  9. Gibberellin-enhanced elongation of inverted Pharbitis nil shoot prevents the release of apical dominance

    Science.gov (United States)

    Prasad, T. K.; Cline, M. G.

    1987-01-01

    Ethylene evolution resulting from the gravity stress of shoot inversion appears to induce the release of apical dominance in Pharbitis nil (L.) by inhibiting elongation of the inverted shoot. It has been previously demonstrated that this shoot inversion release of apical dominance can be prevented by promoting elongation in the inverted shoot via interference with ethylene synthesis or action. In the present study it was shown that apical dominance release can also be prevented by promoting elongation of the inverted shoot via treatment with gibberellic acid (GA3). A synergistic effect was observed when AgNO3, the ethylene action inhibitor, was applied with GA3. Both GA3 and AgNO3 increased ethylene production in the inverted shoot. These results are consistent with the view that it is ethylene-induced inhibition of elongation and not any direct effect of ethylene per se which is responsible for the outgrowth of the highest lateral bud.

  10. Differential Regulation of Apical-basolateral Dendrite Outgrowth by Activity in Hippocampal Neurons

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    Yang eYuan

    2015-08-01

    Full Text Available Hippocampal pyramidal neurons have characteristic dendrite asymmetry, characterized by structurally and functionally distinct apical and basolateral dendrites. The ability of the neuron to generate and maintain dendrite asymmetry is vital, since synaptic inputs received are critically dependent on dendrite architecture. Little is known about the role of neuronal activity in guiding maintainance of dendrite asymmetry. Our data indicate that dendrite asymmetry is established and maintained early during development. Further, our results indicate that cell intrinsic and global alterations of neuronal activity have differential effects on net extension of apical and basolateral dendrites. Thus, apical and basolateral dendrite extension may be independently regulated by cell intrinsic and network neuronal activity during development, suggesting that individual dendrites may have autonomous control over net extension. We propose that regulated individual dendrite extension in response to cell intrinsic and neuronal network activity may allow temporal control of synapse specificity in the developing hippocampus.

  11. Depth of planting and apical dominance on cuttings of red pitayaProfundidade de plantio e dominância apical na estaquia de pitaia vermelha

    Directory of Open Access Journals (Sweden)

    Virna Braga Marques

    2012-12-01

    Full Text Available This study was conducted to evaluate the effects of planting depth and breaking apical dominance on cutting of red pitaya Hylocereus undatus (Haw. Britton & Rose. The experiment was conducted in a randomized block design with four replications in a 2x3 factorial, with two kinds of cuttings (with or without apical dominance, cutting not sectioned and sectioned at 5 cm from the upper portion, and three planting depths (1.0, 5.0, and 10.0 cm and each plot consisted of ten 20 cm long cuttings. After 60 days of planting, counts of the number of shoots were began weekly and 90 days after planting survival rooting, number and length of shoots, dry weight of shoots and root dry mass were evaluated. Data were subjected to analysis of variance and polynomial regression at 5% significance level. The percentage of rooting and the survival were 100% in all treatments. With increasing depth of planting it was observed linear reduction in numbers of shoots, dry weight of shoot and root dry mass. The cladodes with apical dominance had longer shoots. The planting of red pitaya cuttings at 1 cm of planting depth with apical dominance is more suited to the production of nursery plants. O trabalho foi conduzido com o objetivo de avaliar os efeitos da profundidade de plantio e da quebra de dominância apical na estaquia de pitaia vermelha Hylocereus undatus (Haw. Britton & Rose. O experimento foi instalado no delineamento em blocos casualizados, com quatro repetições no esquema fatorial 2x3, sendo dois tipos de estacas (com e sem dominância apical, estacas não seccionadas e seccionadas a 5 cm da porção superior, e três profundidades de plantio (1,0; 5,0; 10,0 cm e cada parcela foi constituída de dez estacas com 20 cm de comprimento. Após 60 dias do plantio, foram iniciadas as avaliações semanais do número de brotações e aos 90 dias após o plantio foram avaliados sobrevivência, enraizamento, número e comprimento de brotações, massa seca das brota

  12. Carrier-mediated ¿-aminobutyric acid transport across the basolateral membrane of human intestinal Caco-2 cell monolayers

    DEFF Research Database (Denmark)

    Nielsen, Carsten Uhd; Carstensen, Mette; Brodin, Birger

    2012-01-01

    and the anticancer prodrug d-aminolevulinic acid across the apical membrane of small intestinal enterocytes. Little is however known about the basolateral transport of these substances. We investigated basolateral transport of GABA in mature Caco-2 cell monolayers using isotope studies. Here we report that, at least...

  13. The apical actin fringe contributes to localized cell wall deposition and polarized growth in the lily pollen tube.

    Science.gov (United States)

    Rounds, Caleb M; Hepler, Peter K; Winship, Lawrence J

    2014-09-01

    In lily (Lilium formosanum) pollen tubes, pectin, a major component of the cell wall, is delivered through regulated exocytosis. The targeted transport and secretion of the pectin-containing vesicles may be controlled by the cortical actin fringe at the pollen tube apex. Here, we address the role of the actin fringe using three different inhibitors of growth: brefeldin A, latrunculin B, and potassium cyanide. Brefeldin A blocks membrane trafficking and inhibits exocytosis in pollen tubes; it also leads to the degradation of the actin fringe and the formation of an aggregate of filamentous actin at the base of the clear zone. Latrunculin B, which depolymerizes filamentous actin, markedly slows growth but allows focused pectin deposition to continue. Of note, the locus of deposition shifts frequently and correlates with changes in the direction of growth. Finally, potassium cyanide, an electron transport chain inhibitor, briefly stops growth while causing the actin fringe to completely disappear. Pectin deposition continues but lacks focus, instead being delivered in a wide arc across the pollen tube tip. These data support a model in which the actin fringe contributes to the focused secretion of pectin to the apical cell wall and, thus, to the polarized growth of the pollen tube. © 2014 American Society of Plant Biologists. All Rights Reserved.

  14. Anoctamin 1 is Apically Expressed on Thyroid Follicular Cells and Contributes to ATP- and Calcium-Activated Iodide Efflux

    Directory of Open Access Journals (Sweden)

    Carmela Iosco

    2014-08-01

    Full Text Available Background/Aims: Iodide efflux from thyroid cells into the follicular lumen is essential for the synthesis of thyroid hormones, however, the pathways mediating this transport have only been partially identified. A calcium-activated pathway of iodide efflux has long been recognized, but its molecular identity unknown. Anoctamin 1 (ANO1 is a calcium-activated chloride channel (CaCC, and this study aims to investigate its contribution to iodide fluxes in thyroid cells. Methods: RT-PCR, immunohistochemistry, and live cell imaging with the fluorescent halide biosensor YFP-H148Q/I152L were used to study the expression, localization and function of ANO1 in thyroid cells. Results: ANO1 mRNA was detected in human thyroid tissue and FRTL-5 thyrocytes, and ANO1 protein was localized to the apical membrane of follicular cells. ATP induced a transient loss of iodide from FRTL-5 cells that was dependent on the mobilization of intracellular calcium, and was inhibited by CaCC/ANO1 inhibitors and siRNA against ANO1. Calcium-activated iodide efflux was also observed in CHO cells over-expressing the Sodium Iodide Symporter (NIS and ANO1. Conclusion: ANO1 in thyrocytes functions as a calcium-activated channel mediating iodide efflux, and may contribute to the rapid delivery of iodide into the follicular lumen for the synthesis of thyroid hormones following activation by calcium-mobilizing stimuli.

  15. Cell competition with normal epithelial cells promotes apical extrusion of transformed cells through metabolic changes.

    Science.gov (United States)

    Kon, Shunsuke; Ishibashi, Kojiro; Katoh, Hiroto; Kitamoto, Sho; Shirai, Takanobu; Tanaka, Shinya; Kajita, Mihoko; Ishikawa, Susumu; Yamauchi, Hajime; Yako, Yuta; Kamasaki, Tomoko; Matsumoto, Tomohiro; Watanabe, Hirotaka; Egami, Riku; Sasaki, Ayana; Nishikawa, Atsuko; Kameda, Ikumi; Maruyama, Takeshi; Narumi, Rika; Morita, Tomoko; Sasaki, Yoshiteru; Enoki, Ryosuke; Honma, Sato; Imamura, Hiromi; Oshima, Masanobu; Soga, Tomoyoshi; Miyazaki, Jun-Ichi; Duchen, Michael R; Nam, Jin-Min; Onodera, Yasuhito; Yoshioka, Shingo; Kikuta, Junichi; Ishii, Masaru; Imajo, Masamichi; Nishida, Eisuke; Fujioka, Yoichiro; Ohba, Yusuke; Sato, Toshiro; Fujita, Yasuyuki

    2017-05-01

    Recent studies have revealed that newly emerging transformed cells are often apically extruded from epithelial tissues. During this process, normal epithelial cells can recognize and actively eliminate transformed cells, a process called epithelial defence against cancer (EDAC). Here, we show that mitochondrial membrane potential is diminished in RasV12-transformed cells when they are surrounded by normal cells. In addition, glucose uptake is elevated, leading to higher lactate production. The mitochondrial dysfunction is driven by upregulation of pyruvate dehydrogenase kinase 4 (PDK4), which positively regulates elimination of RasV12-transformed cells. Furthermore, EDAC from the surrounding normal cells, involving filamin, drives the Warburg-effect-like metabolic alteration. Moreover, using a cell-competition mouse model, we demonstrate that PDK-mediated metabolic changes promote the elimination of RasV12-transformed cells from intestinal epithelia. These data indicate that non-cell-autonomous metabolic modulation is a crucial regulator for cell competition, shedding light on the unexplored events at the initial stage of carcinogenesis.

  16. Influence of Hero Apical instruments on cleaning ovoid-shaped root canals

    Directory of Open Access Journals (Sweden)

    Renato Interliche

    2011-08-01

    Full Text Available The cleaning capacity of Hero 642 nickel-titanium files, complemented by the Hero Apical instruments in flattened roots, was determined by histological analysis, considering the area of action of the instruments on the coronal walls and the presence of remaining debris. Twenty-four single-canal, human mandibular incisors were divided into three groups and prepared as follows: GI, instrumented with Hero 642 NiTi files 30/.06, 25/.06, 20/.06, 25/.06, and 30/.06; GII, instrumented as GI followed by Hero Apical size 30/.06; GIII, instrumented as GI followed by Hero Apical sizes 30/.06 and 30/.08, then returning to 30/.06 with pendulum movements. The apical thirds were prepared for histological processing, analyzed at 40× magnification and the images were examined morphometrically. Statistical analysis showed that GIII presented the best results for removing debris (5.22% ± 4.13, with more contact between the instruments and the root canal walls (19.31% ± 0.15. This differed statistically from GI (14.04% ± 4.96 debris removal, with 42.96% ± 7.11 instrument contact and GII (12.62% ± 5.76 debris removal, with 35.01% ± 0.15 instrument contact. Root canal preparation with Hero 642, complemented by Hero Apical instruments (30/.06 and 30/.08, then re-instrumented with Hero Apical 30/.06 using pendulum movements, was more efficient for debris removal and allowed more contact of the instruments with the root canal walls. GII presented the worst results.

  17. Biofilms and apical periodontitis: study of prevalence and association with clinical and histopathologic findings.

    Science.gov (United States)

    Ricucci, Domenico; Siqueira, José F

    2010-08-01

    This study evaluated the prevalence of bacterial biofilms in untreated and treated root canals of teeth evincing apical periodontitis. The associations of biofilms with clinical conditions, radiographic size, and the histopathologic type of apical periodontitis were also investigated. The material comprised biopsy specimens from 106 (64 untreated and 42 treated) roots of teeth with apical periodontitis. Specimens were obtained by apical surgery or extraction and were processed for histopathologic and histobacteriologic techniques. Bacteria were found in all but one specimen. Overall, intraradicular biofilm arrangements were observed in the apical segment of 77% of the root canals (untreated canals: 80%; treated canals: 74%). Biofilms were also seen covering the walls of ramifications and isthmuses. Bacterial biofilms were visualized in 62% and 82% of the root canals of teeth with small and large radiographic lesions, respectively. All canals with very large lesions harbored intraradicular biofilms. Biofilms were significantly associated with epithelialized lesions (cysts and epithelialized granulomas or abscesses) (p granulomas was 95%, 83%, and 69.5%, respectively. No correlation was found between biofilms and clinical symptoms or sinus tract presence (p > 0.05). Extraradicular biofilms were observed in only 6% of the cases. The overall findings are consistent with acceptable criteria to include apical periodontitis in the set of biofilm-induced diseases. Biofilm morphologic structure varied from case to case and no unique pattern for endodontic infections was identified. Biofilms are more likely to be present in association with longstanding pathologic processes, including large lesions and cysts. Copyright 2010 American Association of Endodontists. Published by Elsevier Inc. All rights reserved.

  18. Sequential development of apical-basal and planar polarities in aggregating epitheliomuscular cells of Hydra.

    Science.gov (United States)

    Seybold, Anna; Salvenmoser, Willi; Hobmayer, Bert

    2016-04-01

    Apical-basal and planar cell polarities are hallmarks of metazoan epithelia required to separate internal and external environments and to regulate trans- and intracellular transport, cytoskeletal organization, and morphogenesis. Mechanisms of cell polarization have been intensively studied in bilaterian model organisms, particularly in early embryos and cultured cells, while cell polarity in pre-bilaterian tissues is poorly understood. Here, we have studied apical-basal and planar polarization in regenerating (aggregating) clusters of epitheliomuscular cells of Hydra, a simple representative of the ancestral, pre-bilaterian phylum Cnidaria. Immediately after dissociation, single epitheliomuscular cells do not exhibit cellular polarity, but they polarize de novo during aggregation. Reestablishment of the Hydra-specific epithelial bilayer is a result of short-range cell sorting. In the early phase of aggregation, apical-basal polarization starts with an enlargement of the epithelial apical-basal diameter and by the development of belt-like apical septate junctions. Specification of the basal pole of epithelial cells occurs shortly later and is linked to synthesis of mesoglea, development of hemidesmosome-like junctions, and formation of desmosome-like junctions connecting the basal myonemes of neighbouring cells. Planar polarization starts, while apical-basal polarization is already ongoing. It is executed gradually starting with cell-autonomous formation, parallelization, and condensation of myonemes at the basal end of each epithelial cell and continuing with a final planar alignment of epitheliomuscular cells at the tissue level. Our findings reveal that epithelial polarization in Hydra aggregates occurs in defined steps well accessible by histological and ultrastructural techniques and they will provide a basis for future molecular studies. Copyright © 2016 Elsevier Inc. All rights reserved.

  19. Frecuencia de periodontitis apical en tratamientos endodónticos de pregrado

    OpenAIRE

    León, P.; Ilabaca, M.J.; Alcota, M.; González, F.E.

    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...

  20. Criterios Clínicos y Radiológicos de la Periodontitis Apical.

    OpenAIRE

    Jiménez Planas, Amparo

    2017-01-01

    Se realiza un trabajo con 271 casos diagnosticados de periodontitis apical, clínica o radiológicamente, en los que se valoran parámetros clínicos y radiológicos que nos permitan delimitar las diferentes formas clínicas de este proceso. se propone, como consecuencia de los resultados, una clasificación clara, simple, coherente y evolutiva de la periodontitis apical y se hacen diversas consideraciones sobre la nomenclatura, etiología, cl...

  1. Reestablishing the Function and Esthetics in Traumatized Permanent Teeth with Large Apical Lesion

    Directory of Open Access Journals (Sweden)

    Alexandra Rubin Cocco

    2016-01-01

    Full Text Available Dental trauma is a challenge for dental integrity and can lead to pulp necrosis. The clinical case reports the diagnosis of a maxillary right central incisor traumatized and its multidisciplinary treatment. Calcium hydroxide material was used to perform the processing apexification. An apical surgery was carried out to remove the apical periodontitis and to return the aesthetics to the patient; internal and external tooth whitening in maxillary right central incisor was performed. We conclude that surgery associated with the root filling in the central incisor led to a successful completion. Moreover, it is of utmost importance to demonstrate the interaction between the various areas of dentistry.

  2. Periimplantitis apical - presentación de serie de 11 casos clínicos

    OpenAIRE

    Stavaru Marinescu, Bogdan; Naval Gías, Luis; Herrera Calvo, Gonzalo

    2015-01-01

    Objetivos: Evaluar el protocolo quirúrgico y discutir los posibles factores predisponentes de la periimplantitis apical. Material y método: En el presente trabajo, se planteó un estudio descriptivo retrospectivo analizando una serie de 11 casos clínicos de periimplantitis apical diagnosticados y tratados en el ámbito del Hospital de La Princesa (Madrid) y la Clínica Universidad de Navarra (Pamplona) entre 2002 y 2013. Los pacientes sintomáticos fueron tratados mediante legrado de la zona c...

  3. Safety and immunogenicity of multi-antigen AMA1-based vaccines formulated with CoVaccine HT™ and Montanide ISA 51 in rhesus macaques

    Directory of Open Access Journals (Sweden)

    Walraven Vanessa

    2011-07-01

    Full Text Available Abstract Background Increasing the breadth of the functional antibody response through immunization with Plasmodium falciparum apical membrane antigen 1 (PfAMA1 multi-allele vaccine formulations has been demonstrated in several rodent and rabbit studies. This study assesses the safety and immunogenicity of three PfAMA1 Diversity-Covering (DiCo vaccine candidates formulated as an equimolar mixture (DiCo mix in CoVaccine HT™ or Montanide ISA 51, as well as that of a PfAMA1-MSP119 fusion protein formulated in Montanide ISA 51. Methods Vaccine safety in rhesus macaques was monitored by animal behaviour observation and assessment of organ and systemic functions through clinical chemistry and haematology measurements. The immunogenicity of vaccine formulations was assessed by enzyme-linked immunosorbent assays and in vitro parasite growth inhibition assays with three culture-adapted P. falciparum strains. Results These data show that both adjuvants were well tolerated with only transient changes in a few of the chemical and haematological parameters measured. DiCo mix formulated in CoVaccine HT™ proved immunologically and functionally superior to the same candidate formulated in Montanide ISA 51. Immunological data from the fusion protein candidate was however difficult to interpret as four out of six immunized animals were non-responsive for unknown reasons. Conclusions The study highlights the safety and immunological benefits of DiCo mix as a potential human vaccine against blood stage malaria, especially when formulated in CoVaccine HT™, and adds to the accumulating data on the specificity broadening effects of DiCo mix.

  4. Comparison of Milk Fat Globule Membrane (MFGM) Proteins of Chianina and Holstein Cattle Breed Milk Samples Through Proteomics Methods

    OpenAIRE

    Lorraine Pariset; Silvia Bongiorni; Alessio Valentini; Lello Zolla; Leonardo Murgiano; Anna Maria Timperio

    2009-01-01

    Identification of proteins involved in milk production is important to understand the biology of lactation. Many studies have advanced the understanding of mammary function and milk secretion, but the critical molecular mechanisms implicated in milk fat secretion is still incomplete. Milk Fat Globules are secreted from the apical surface of the mammary cells, surrounded by a thin membrane bilayer, the Milk Fat Globule Membrane (MFGM), formed by proteins which have been suggested to be cholest...

  5. Identification and characterization of Neospora caninum tachyzoite antigens useful for diagnosis of neosporosis.

    Science.gov (United States)

    Bjerkas, I; Jenkins, M C; Dubey, J P

    1994-01-01

    The purpose of the present study was to identify antigens of the protozoan Neospora caninum that could be useful for the diagnosis of neosporosis in domestic animals. As revealed by immunoblotting, immune sera from a wide range of animal species exhibited a similar recognition pattern of four major and several minor N. caninum antigens. In contrast to preinoculation sera, all tested immune sera recognized nonreduced immunodominant 17-, 29-, 30-, and 27-kDa antigens. A 46-kDa protein which showed faint recognition by preimmune sera also exhibited a strong response by immune sera. Immunolocalization of the four immunodominant N. caninum antigens was investigated by immunogold electron microscopy using monospecific polyclonal antisera. The 17-kDa antigen appears to be associated with the body part of the rhoptries, while the 29- and 30-kDa antigens were associated with the dense granules, network, and limiting membrane of the parasitophorous vacuole. Studies were also conducted to compare antibody responses to N. caninum and the related protozoan Toxoplasma gondii. Although N. caninum and T. gondii (RH strain) tachyzoites shared a few cross-reacting antigens, the immunodominant antigens of both parasites were not recognized by heterologous sera. Also, immunogold staining with rabbit anti-Neospora hyperimmune serum exhibited almost no labeling of external membranes of Neospora tachyzoites compared with the very marked labeling seen when Toxoplasma tachyzoites (RH strain) were incubated with rabbit anti-Toxoplasma hyperimmune serum. These unique antigenic differences should be useful in developing a diagnostic assay for N. caninum. Images PMID:7496948

  6. 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.

  7. Localization of the terminal steps of O-antigen synthesis in Salmonella typhimurium

    International Nuclear Information System (INIS)

    McGrath, B.C.; Osborn, M.J.

    1991-01-01

    Previous immunoelectron microscopic studies have shown that both the final intermediate in O-antigen synthesis, undecaprenol-linked O polymer, and newly synthesized O-antigenic lipopolysaccharide are localized to the periplasmic face of the inner membrane. In vivo pulse-chase experiments now provide further evidence that attachment of O antigen to core lipopolysaccharide, as well as polymerization of O-specific polysaccharide chains, takes place at the periplasmic face of the membrane. Mutants doubly conditional in lipopolysaccharide synthesis [kdsA(Ts) pmi] were constructed in which synthesis of core lipopolysaccharide and O antigen are temperature sensitive and mannose dependent, respectively. Periplasmic orientation of O antigen:core lipopolysaccharide ligase was established by experiments showing rapid chase of undecaprenol-linked O polymer, previously accumulated at 42 degrees C in the absence of core synthesis, into lipopolysaccharide following resumption of core formation at 30 degrees C. In addition, chase of the monomeric O-specific tetrasaccharide unit into lipopolysaccharide was found in similar experiments in an O-polymerase-negative [rfc kdsA(Ts) pmi] mutant, suggesting that polymerization of O chains also occurs at the external face of the inner membrane

  8. Evaluation and Comparison of the Position of the Apical Constriction in Single-root and Multiple-root Teeth

    Directory of Open Access Journals (Sweden)

    Alireza Farhad

    2017-12-01

    Full Text Available Introduction: Precise knowledge of the location of the apical constriction is essential to root canal treatment and long-term prognosis. Considering the differences in the apical constriction and size of the roots in single- and multiple-root teeth in various races, examination and comparison of the location of the apical constriction in single-root and multiple-root teeth are of paramount importance. The present studies aimed to measure and compare the distance of the apical constriction from the apical foramen and anatomical apex in single-root and multiple-root teeth. Materials and Methods: In this cross-sectional study, 60 roots of single-rooted teeth and 60 roots of multiple-rooted teeth were collected from the patients referring to the health centers in Isfahan, Iran. After cleansing and disinfecting the surface of the roots, the surface of the teeth was washed with hypochlorite. Based on the direction of the apical foramen, a longitudinal cut was made in the same direction, and the roots were examined microscopically at the magnification of 25. Following that, the distance of the apical constriction from the apical foramen and anatomical apex was measured using a digital camera. In addition, mean and standard deviation of the obtained distance values were determined. Distances in the single-root and multiple-root teeth were compared using independent t-test, at the significance level of Results: Mean distance between the apical constriction and apical foramen was 0.86±0.33 mm in the single-root teeth and 0.072±0.27 mm in the multiple-root teeth. Mean distance between the apical constriction and anatomical apex was 1.14±0.36 mm in the single-root teeth and 1.03±0.36 mm in the multiple-root teeth. Moreover, the results of independent t-test showed the distance of the apical constriction from the apical foramen to be significant between single-root and multiple-rooted teeth (P=0.013. However, the distance between the apical constriction

  9. Predicting placebo response in adolescents with major depressive disorder: The Adolescent Placebo Impact Composite Score (APICS).

    Science.gov (United States)

    Nakonezny, Paul A; Mayes, Taryn L; Byerly, Matthew J; Emslie, Graham J

    2015-09-01

    The aim of this study was to construct a composite scoring system to predict the probability of placebo response in adolescents with Major Depressive Disorder (MDD). Participants of the current study were 151 adolescents (aged 12-17 years) who were randomized to the placebo arm (placebo transdermal patches) of a randomized controlled trial (RCT) comparing the selegiline transdermal patch with placebo (DelBello et al., 2014). The primary outcome of response was defined as a CGI-I score of 1 or 2 (very much or much improved) at week 12 (study-end) or exit. As a first step, a multiple logistic mixed model was used to estimate the odds of placebo response from each predictor in the model, including age, CDRS-R total at baseline (depressive symptom severity), history of recurrent depression (yes vs. no), sex (female vs. male), and race (non-Caucasian vs. Caucasian). On the basis of the initial logistic mixed model analysis, we then constructed an Adolescent Placebo Impact Composite Score (APICS) that became the sole predictor in a re-specified Bayesian logistic regression model to estimate the probability of placebo response. Finally, the AUC for the APICS was tested against a nominal area of 0.50 to evaluate how well the APICS discriminated placebo response status. Among the 151 adolescents, with a mean age of 14.6 years (SD = 1.6) and a mean baseline CDRS-R total of 60.6 (SD = 12.1), 68.2% were females, 50.3% was Caucasian, and 39.7% had a history of recurrent depression. Placebo response rate was 58.3%. Based on the logistic mixed model, the re-specified equation with the highest discriminatory ability to estimate the probability of placebo response was APICS = age + (0.32 × CDRS-R Total at baseline) + (-2.85 × if female) + (-5.50 × if history of recurrent depression) + (-5.85 × if non-Caucasian). The AUC for this model was 0.59 (p = .049). Within a Bayesian decision-theoretic framework, in 95.5% of the time, the 10,000 posterior Monte Carlo samples suggested

  10. Nonstripping "Rainbow" and Multiple Antigen Detection (MAD) Western Blotting.

    Science.gov (United States)

    Krajewski, Stan Stanislaw; Tsukamoto, Michelle M; Huang, Xianshu; Krajewski, Sebastian B

    2015-01-01

    A variation of immunoblotting method (the "Rainbow Western"), permits sequential detection of multiple antigens (MAD) on a single protein blot without stripping off prior antibodies. Because no stripping is involved, immobilized proteins are not lost from the membrane, thus allowing for multiple reprobings of the same membrane with different primary antibodies (≥12), retaining strong signal intensities for all sequential antibody probings. The procedure utilizes horseradish peroxidase (HRPase)-based detection with both a chemiluminescent and colorimetric substrate. Initial incubation of the blot with secondary antibody followed by colorimetric development prior to probing the blot with primary antibodies markedly reduces background in ECL-based detection procedures and permits sequential use of antibodies derived from a single species. In the "Rainbow Western," four different HRPase-colorimetric substrates that produce black, brown, red, and green colors are employed sequentially for detection and simultaneous display of four different antigens on the same membrane. By allowing large amounts of data to be obtained from a single blot, the MAD-immunoblotting and Rainbow Western methods have the potential for researchers to compare the expression of several proteins within a single biological sample. Both techniques could be particularly valuable for analysis of cellular populations that are difficult to isolate in large numbers or of clinical specimens where the amounts of protein samples is minute or only available on a one-time basis.

  11. Human sensitization to Ganoderma antigen.

    Science.gov (United States)

    Tarlo, S M; Bell, B; Srinivasan, J; Dolovich, J; Hargreave, F E

    1979-07-01

    Continuous air sampling with a Hirst volumetric spore trap over 3 yr has identified basidiospores of Ganoderma applanatum, a bracket fungus, as the most numerous fungal spores in two southern Ontario locations. The particle size is small and the calculated total spore mass approximates that of the spores of Cladosporium and Alternaria. Extracts of Ganoderma applanatum bracket fungus and spores in w/v, 1:10 concentration were prepared after collection of samples of the fungus from local woods. Skin prick tests with the extracts were performed in 294 consecutive children and adults attending two chest/allergy clinics. Of these patients, 182 (61.9%) reacted to 1 or more of the common inhalant allergen extracts and 24 (8.2%) reacted to Ganoderma antigen. There was no consistent relationship between reactivity to Ganoderma antigen and any of the common inhaled allergens. IgE-dependent sensitization to Ganoderma was confirmed by the radioallergosorbent test (RAST). Rabbit antisera to Ganoderma antigen preparations did not appear to cross-react with preparations of the various clinically important allergens. The findings indicate that Ganoderma antigen is commonly encountered, can induce human sensitization, and has unique antigenicity among common allergens of clinical importance.

  12. Chitosan cushioned phospholipid membrane and its application in imaging ellipsometry based-biosensor

    International Nuclear Information System (INIS)

    Zhang Yibang; Chen Yanyan; Jin Gang

    2011-01-01

    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.

  13. Specific Lipids Modulate the Transporter Associated with Antigen Processing (TAP)*

    Science.gov (United States)

    Schölz, Christian; Parcej, David; Ejsing, Christer S.; Robenek, Horst; Urbatsch, Ina L.; Tampé, Robert

    2011-01-01

    The transporter associated with antigen processing (TAP) plays a key role in adaptive immunity by translocating proteasomal degradation products from the cytosol into the endoplasmic reticulum lumen for subsequent loading onto major histocompatibility (MHC) class I molecules. For functional and structural analysis of this ATP-binding cassette complex, we established the overexpression of TAP in the methylotrophic yeast Pichia pastoris. Screening of optimal solubilization and purification conditions allowed the isolation of the heterodimeric transport complex, yielding 30 mg of TAP/liter of culture. Detailed analysis of TAP function in the membrane, solubilized, purified, and reconstituted states revealed a direct influence of the native lipid environment on activity. TAP-associated phospholipids, essential for function, were profiled by liquid chromatography Fourier transform mass spectrometry. The antigen translocation activity is stimulated by phosphatidylinositol and -ethanolamine, whereas cholesterol has a negative effect on TAP activity. PMID:21357424

  14. 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

  15. Search for Mycobacterium avium Subspecies paratuberculosis Antigens for the Diagnosis of Paratuberculosis

    NARCIS (Netherlands)

    Mon, M.L.; Vale, M.; Baschetti, G.; Alvarado Pinedo, F.; Gioffre, A.; Traveria, G.; Willemsen, P.; Bakker, D.; Romano, M.I.

    2012-01-01

    The aim of this study was to evaluate a wide panel of antigens of Mycobacterium avium subsp. paratuberculosis (MAP) to select candidates for the diagnosis of paratuberculosis (PTB). A total of 54 recombinant proteins were spotted onto nitrocellulose membranes and exposed to sera from animals with

  16. Role of an apical K,Cl cotransporter in urine formation by renal tubules of the yellow fever mosquito (Aedes aegypti).

    Science.gov (United States)

    Piermarini, Peter M; Hine, Rebecca M; Schepel, Matthew; Miyauchi, Jeremy; Beyenbach, Klaus W

    2011-11-01

    The K,Cl cotransporters (KCCs) of the SLC12 superfamily play critical roles in the regulation of cell volume, concentrations of intracellular Cl(-), and epithelial transport in vertebrate tissues. To date, the role(s) of KCCs in the renal functions of mosquitoes and other insects is less clear. In the present study, we sought molecular and functional evidence for the presence of a KCC in renal (Malpighian) tubules of the mosquito Aedes aegypti. Using RT-PCR on Aedes Malpighian tubules, we identified five alternatively spliced partial cDNAs that encode putative SLC12-like KCCs. The majority transcript is AeKCC1-A(1); its full-length cDNA was cloned. After expression of the AeKCC1-A protein in Xenopus oocytes, the Cl(-)-dependent uptake of (86)Rb(+) is 1) activated by 1 mM N-ethylmaleimide and cell swelling, 2) blocked by 100 μM dihydroindenyloxyalkanoic acid (DIOA), and 3) dependent upon N-glycosylation of AeKCC1-A. In Aedes Malpighian tubules, AeKCC1 immunoreactivity localizes to the apical brush border of principal cells, which are the predominant cell type in the epithelium. In vitro physiological assays of Malpighian tubules show that peritubular DIOA (10 μM): 1) significantly reduces both the control and diuretic rates of transepithelial fluid secretion and 2) has negligible effects on the membrane voltage and input resistance of principal cells. Taken together, the above observations indicate the presence of a KCC in the apical membrane of principal cells where it participates in a major electroneutral transport pathway for the transepithelial secretion of fluid in this highly electrogenic epithelium.

  17. Freeze-fracture studies of photoreceptor membranes: new observations bearing upon the distribution of cholesterol.

    Science.gov (United States)

    Andrews, L D; Cohen, A I

    1983-09-01

    We performed electron microscopy of replicas from freeze-fractured retinas exposed during or after fixation to the cholesterol-binding antibiotic, filipin. We observed characteristic filipin-induced perturbations throughout the disk and plasma membranes of retinal rod outer segments of various species. It is evident that a prolonged exposure to filipin in fixative enhances rather than reduces presumptive cholesterol detection in the vertebrate photoreceptor cell. In agreement with the pattern seen in our previous study (Andrews, L.D., and A. I. Cohen, 1979, J. Cell Biol., 81:215-228), filipin-binding in membranes exhibiting particle-free patches seemed largely confined to these patches. Favorably fractured photoreceptors exhibited marked filipin-binding in apical inner segment plasma membrane topologically confluent with and proximate to the outer segment plasma membrane, which was comparatively free of filipin binding. A possible boundary between these differing membrane domains was suggested in a number of replicas exhibiting lower filipin binding to the apical plasma membrane of the inner segment in the area surrounding the cilium. This area contains a structure (Andrews, L. D., 1982, Freeze-fracture studies of vertebrate photoreceptors, In Structure of the Eye, J. G. Hollyfield and E. Acosta Vidrio, editors, Elsevier/North-Holland, New York, 11-23) that resembles the active zones of the nerve terminals for the frog neuromuscular junction. These observations lead us to hypothesize that these structures may function to direct vesicle fusion to occur near them, in a domain of membrane more closely resembling outer than inner segment plasma membrane. The above evidence supports the views that (a) all disk membranes contain cholesterol, but the particle-free patches present in some disks trap cholesterol from contiguous particulate membrane regions; (b) contiguous inner and outer segment membranes may greatly differ in cholesterol content; and (c) the suggested

  18. A unique approach to demonstrating that apical bud temperature specifically determines leaf initiation rate in the dicot Cucumis sativus

    NARCIS (Netherlands)

    Savvides, Andreas; Dieleman, Anja; Ieperen, van Wim; Marcelis, Leo F.M.

    2016-01-01

    Main conclusion: Leaf initiation rate is largely determined by the apical bud temperature even when apical bud temperature largely deviates from the temperature of other plant organs.We have long known that the rate of leaf initiation (LIR) is highly sensitive to temperature, but previous studies

  19. Alterations in apical dendrite bundling in the somatosensory cortex of 5-HT3A receptor knockout mice.

    NARCIS (Netherlands)

    Smit-Rigter, L.A.; Wadman, W.J.; van Hooft, J.A.

    2011-01-01

    In various species and areas of the cerebral cortex, apical dendrites of pyramidal neurons form clusters which extend through several layers of the cortex also known as dendritic bundles. Previously, it has been shown that 5-HT3A receptor knockout mice show hypercomplex apical dendrites of cortical

  20. Characterisation of Sarcoptes scabiei antigens.

    Science.gov (United States)

    Hejduk, Gloria; Hofstätter, Katja; Löwenstein, Michael; Peschke, Roman; Miller, Ingrid; Joachim, Anja

    2011-02-01

    In pig herds, the status of Sarcoptes scabiei infections is routinely monitored by serodiagnosis. Crude antigen for ELISA is usually prepared from S. scabiei var. canis or other variations and may lead to variations in the outcome of different tests, making assay standardisation difficult. This study was performed to investigate the antigen profiles of S. scabiei, including differences between hydrophilic and more hydrophobic protein fractions, by Western blotting with sera from pigs with defined infection status. Potential cross-reactivity among S. scabiei (var. canis, suis and bovis), Dermatophagoides farinae and Tyrophagus putrescentiae was also analysed. Hydrophobic S. scabiei antigens were detectable in the range of 40-50 kDa, whilst the hydrophilic fraction showed no specific antigenicity. In the hydrophobic fractions of D. farinae and T. putrescentiae, two major protein fractions in a similar size range could be identified, but no cross-reactivity with Sarcoptes-positive sera was detectable. However, examination of the hydrophilic fractions revealed cross-reactivity between Sarcoptes-positive sera and both the house dust mite and the storage mite in the range of 115 and 28/38 kDa. Specific bands in the same range (42 and 48 kDa) could be detected in blots from hydrophobic fractions of all three tested variations of S. scabiei (var. canis, bovis and suis). These results show that there are considerable differences in mange antibody reactivity, including reactions with proteins from free-living mites, which may interfere with tests based on hydrophilic antigens. Further refinement of antigen and the use of specific hydrophobic proteins could improve ELISA performance and standardisation.

  1. 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.

  2. Expression of Podoplanin in the Mouse Tooth Germ and Apical Bud Cells

    Science.gov (United States)

    Sawa, Yoshihiko; Iwasawa, Kana; Ishikawa, Hiroyuki

    2008-01-01

    This study was designed to investigate the distribution of cells expressing podoplanin in the mouse tooth bud. Podoplanin expression was detected in enamel epithelia of the cervical loop at cell-cell contacts strongly, and weakly on the loosely aggregated stellate reticulum in the center and the neighboring stratum intermedium. Odontoblasts exhibited intense podoplanin expression at the junction with predentin while no expression was detected in the enamel organ containing ameloblasts. These results suggest that proliferating inner and outer enamel epithelia express podoplanin but that the expression is suppressed in the differentiated epithelia containing ameloblasts. On the other hand the podoplanin expression occurs in the differentiating odontoblasts and the expression is sustained in differentiated odontoblasts, indicating that odontoblasts have the strong ability to express podoplanin. In cultured apical bud cells podoplanin was detected at cell-cell contacts. In real-time PCR analysis the amount of podoplanin mRNA of the apical buds was 2-fold compared with the amount of kidney used as a positive control. These findings indicate that apical bud cells have the strong ability to express the podoplanin gene. Podoplanin is a mucin-type glycoprotein negatively charged by extensive O-glycosylation and a high content of sialic acid, which expresses the adhesive property. The podoplanin may contribute to form odontoblastic fiber or function as the anchorage to the tooth development and in proliferating epithelial cells of cervical loop and apical bud. PMID:18989465

  3. 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.

  4. Electric current affects the rate of development in isolated apical parts of rape in vitro

    Czech Academy of Sciences Publication Activity Database

    Filek, M.; Koscielniak, J.; Marcińska, I.; Krekule, Jan; Macháčková, Ivana; Dubert, F.

    2006-01-01

    Roč. 50, č. 3 (2006), s. 465-468 ISSN 0006-3134 Institutional research plan: CEZ:AV0Z5038910 Keywords : apical meristem differentiation * flowering factors * vernalization Subject RIV: EF - Botanics Impact factor: 1.198, year: 2006

  5. Oxidative Stress in the Local and Systemic Events of Apical Periodontitis

    Directory of Open Access Journals (Sweden)

    Patricia Hernández-Ríos

    2017-11-01

    Full Text Available Oxidative stress is involved in the pathogenesis of a variety of inflammatory disorders. Apical periodontitis (AP usually results in the formation of an osteolytic apical lesion (AL caused by the immune response to endodontic infection. Reactive oxygen species (ROS produced by phagocytic cells in response to bacterial challenge represent an important host defense mechanism, but disturbed redox balance results in tissue injury. This mini review focuses on the role of oxidative stress in the local and associated systemic events in chronic apical periodontitis. During endodontic infection, ligation of Toll-like receptors (TLRs on phagocytes' surface triggers activation, phagocytosis, synthesis of ROS, activation of humoral and cellular responses, and production of inflammatory mediators, such as, cytokines and matrix metalloproteinases (MMPs. The increment in ROS perturbs the normal redox balance and shifts cells into a state of oxidative stress. ROS induce molecular damage and disturbed redox signaling, that result in the loss of bone homeostasis, increased pro-inflammatory mediators, and MMP overexpression and activation, leading to apical tissue breakdown. On the other hand, oxidative stress has been strongly involved in the pathogenesis of atherosclerosis, where a chronic inflammatory process develops in the arterial wall. Chronic AP is associated with an increased risk of cardiovascular diseases (CVD and especially atherogenesis. The potential mechanisms linking these diseases are also discussed.

  6. Clinical sustained uniform ventricular tachycardia in hypertrophic cardiomyopathy: association with left ventricular apical aneurysm.

    OpenAIRE

    Alfonso, F; Frenneaux, M P; McKenna, W J

    1989-01-01

    Of 51 patients with hypertrophic cardiomyopathy who had episodes of ventricular tachycardia detected during ambulatory electrocardiographic monitoring only two had clinical sustained uniform ventricular tachycardia that required medical treatment because of worsening symptoms. In both patients the arrhythmia was associated with the uncommon finding of an apical aneurysm with angiographically normal coronary arteries.

  7. Gingival recession following apical surgery in the esthetic zone: a clinical study with 70 cases

    DEFF Research Database (Denmark)

    von Arx, Thomas; Salvi, Giovanni E; Janner, Simone

    2009-01-01

    The present study evaluated gingival recession 1 year following apical surgery of 70 maxillary anterior teeth (central and lateral incisors, canines, and first premolars). A visual assessment of the mid-facial aspect of the gingival level and of papillary heights of treated teeth was carried out...

  8. 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

  9. 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

  10. Coordination of growth in root and shoot apices by AIL/PLT transcription factors

    NARCIS (Netherlands)

    Scheres, Ben; Krizek, Beth A.

    2018-01-01

    Growth at the root tip and organ generation at the shoot tip depend on the proper functioning of apical meristems and the transitioning of meristematic cell descendants from a proliferating state to cell elongation and differentiation. Members of the AINTEGUMENTA-LIKE/PLETHORA (AIL/PLT)

  11. 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.

  12. RECIPROC VERSUS TWISTED FILE FOR ROOT CANAL FILLING REMOVAL:ASSESSMENT OF APICALLY EXTRUDED DEBRIS*

    Directory of Open Access Journals (Sweden)

    Demet ALTUNBAŞ

    2016-04-01

    Full Text Available Purpose: The aim of this study was to evaluate the amount of apically extruded debris during endodontic retreatment with different file systems. Materials and Methods: Sixty extracted human mandibular premolar teeth were used in this study. Root canals of the teeth were instrumented and filled before being randomly assigned to three groups. Guttapercha was removed using the Reciproc system, the Twisted File system (TF, and Hedström-files (H-file. Apically extruded debris was collected and dried in pre-weighed Eppendorf tubes. The amount of extruded debris was assessed with an electronic balance. Data were statistically analyzed using one-way ANOVA, Kruskal-Wallis, and Mann-Whitney U tests. Results: The Reciproc and TF systems extruded significantly less debris than the H-file (p<0.05. However, no significant difference was found between the Reciproc and TF systems. Conclusion: All tested file systems caused apical extrusion of debris. Both the rotary file (TF and the reciprocating single-file (Reciproc systems were associated with less apical extrusion compared with the H-file.

  13. 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

    2015-10-01

    This study evaluated the frequency of root resorption during the orthodontic treatment with Herbst appliance by Cone Beam Computed Tomography (CBCT). 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%. 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. CBCT three-dimensional evaluation showed association between Herbst appliance and minimal apical root resorption, mostly in the anchoring teeth, without clinical significance.

  14. Evaluation of the amount of apically extruded debris using Mtwo and ...

    African Journals Online (AJOL)

    ajl yemi

    2011-12-26

    Dec 26, 2011 ... Extrusion of debris into the periradicular tissue during endodontic treatment is one of the main causes of postoperative flare-ups. The aim of this study was to compare the amount of apically extruded debris using Mtwo instruments with single length technique and RaCe system using crown-down approach.

  15. Identification of apical vertebra for grading of idiopathic scoliosis using image processing.

    Science.gov (United States)

    Anitha, H; Prabhu, G K

    2012-02-01

    Scoliosis is a 3-D deformity of spinal column, characterized by both lateral curvature and vertebral rotation. The disease can be caused by congenital, developmental, or degenerative problems; but most cases of scoliosis actually have no known cause, and this is known as idiopathic scoliosis. Vertebral rotation has become increasingly prominent in the study of scoliosis and the most deformed vertebra is named as apical vertebra. Apical vertebral deformity demonstrates significance in both preoperative and postoperative assessment, providing better appreciation of the impact of bracing or surgical interventions. Precise measurement of apical vertebral rotation in terms of grading is most valuable for the determination of reference value in normal and pathological conditions for better understanding of scoliosis. Routine quantitative evaluation of vertebral rotation is difficult and error prone due to limitations of observer characteristic and specific imaging property. This paper proposes automatic identification of the apical vertebra and its parameter that depends on the objective criteria of measurement using active contour models. The proposed technique is more accurate and is a reliable measurement compared to manual and computer-assisted system.

  16. Role of apical oxygen in 2-1-4 electron-doped superconductors

    Czech Academy of Sciences Publication Activity Database

    Richard, P.; Riou, G.; Jandl, S.; Poirier, M.; Furnier, P.; Nekvasil, Vladimír; Diviš, M.

    408-410, - (2004), s. 830-831 ISSN 0921-4534 R&D Projects: GA ČR GA202/03/0552 Institutional research plan: CEZ:AV0Z1010914 Keywords : Nd 2-x Ce x CuO 4 * crystal-field * apical oxygen * HTSC Subject RIV: BM - Solid Matter Physics ; Magnetism Impact factor: 1.072, year: 2004

  17. Three Arabidopsis AIL/PLT genes act in combination to regulate shoot apical meristem function.

    Science.gov (United States)

    Mudunkothge, Janaki S; Krizek, Beth A

    2012-07-01

    The shoot apical meristem, a small dome-shaped structure at the shoot apex, is responsible for the initiation of all post-embryonic shoot organs. Pluripotent stem cells within the meristem replenish themselves and provide daughter cells that become incorporated into lateral organ primordia around the meristem periphery. We have identified three novel regulators of shoot apical meristem activity in Arabidopsis thaliana that encode related AIL/PLT transcription factors: AINTEGUMENTA (ANT), AINTEGUMENTA-LIKE6 (AIL6)/PLETHORA3 (PLT3) and AINTEGUMENTA-LIKE7 (AIL7)/PLETHORA7 (PLT7). Loss of these genes results in plants that initiate only a few leaves prior to termination of shoot apical meristem activity. In 7-day-old ant ail6 ail7 seedlings, we observed reduced cell division in the meristem region, differentiation of meristematic cells and altered expression of the meristem regulators WUSCHEL (WUS), CLAVATA3 (CLV3) and SHOOT MERISTEMLESS (STM). Genetic experiments suggest that these three AIL genes do not act specifically in either the WUS/CLV or STM pathway regulating meristem function. Furthermore, these studies indicate that ANT, AIL6 and AIL7 have distinct functions within the meristem rather than acting in a strictly redundant manner. Our study thus identifies three new genes whose distinct functions are together required for continuous shoot apical meristem function. © 2012 The Authors. The Plant Journal © 2012 Blackwell Publishing Ltd.

  18. 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.

  19. 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,

  20. 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 wer...

  1. Apically extruded dentin debris by reciprocating single-file and multi-file rotary system.

    Science.gov (United States)

    De-Deus, Gustavo; Neves, Aline; Silva, Emmanuel João; Mendonça, Thais Accorsi; Lourenço, Caroline; Calixto, Camila; Lima, Edson Jorge Moreira

    2015-03-01

    This study aims to evaluate the apical extrusion of debris by the two reciprocating single-file systems: WaveOne and Reciproc. Conventional multi-file rotary system was used as a reference for comparison. The hypotheses tested were (i) the reciprocating single-file systems extrude more than conventional multi-file rotary system and (ii) the reciprocating single-file systems extrude similar amounts of dentin debris. After solid selection criteria, 80 mesial roots of lower molars were included in the present study. The use of four different instrumentation techniques resulted in four groups (n = 20): G1 (hand-file technique), G2 (ProTaper), G3 (WaveOne), and G4 (Reciproc). The apparatus used to evaluate the collection of apically extruded debris was typical double-chamber collector. Statistical analysis was performed for multiple comparisons. No significant difference was found in the amount of the debris extruded between the two reciprocating systems. In contrast, conventional multi-file rotary system group extruded significantly more debris than both reciprocating groups. Hand instrumentation group extruded significantly more debris than all other groups. The present results yielded favorable input for both reciprocation single-file systems, inasmuch as they showed an improved control of apically extruded debris. Apical extrusion of debris has been studied extensively because of its clinical relevance, particularly since it may cause flare-ups, originated by the introduction of bacteria, pulpal tissue, and irrigating solutions into the periapical tissues.

  2. Reciproc versus Twisted file for root canal filling removal: assessment of apically extruded debris.

    Science.gov (United States)

    Altunbas, Demet; Kutuk, Betul; Toyoglu, Mustafa; Kutlu, Gizem; Kustarci, Alper; Er, Kursat

    2016-01-01

    The aim of this study was to evaluate the amount of apically extruded debris during endodontic retreatment with different file systems. Sixty extracted human mandibular premolar teeth were used in this study. Root canals of the teeth were instrumented and filled before being randomly assigned to three groups. Guttapercha was removed using the Reciproc system, the Twisted File system (TF), and Hedström-files (H-file). Apically extruded debris was collected and dried in pre-weighed Eppendorf tubes. The amount of extruded debris was assessed with an electronic balance. Data were statistically analyzed using one-way ANOVA, Kruskal-Wallis, and Mann-Whitney U tests. The Reciproc and TF systems extruded significantly less debris than the H-file (p<0.05). However, no significant difference was found between the Reciproc and TF systems. All tested file systems caused apical extrusion of debris. Both the rotary file (TF) and the reciprocating single-file (Reciproc) systems were associated with less apical extrusion compared with the H-file.

  3. Evaluation of apically extruded debris and irrigant using different file systems.

    Science.gov (United States)

    Kütük, B; Akpınar, K E; Altunbaş, D

    2017-12-01

    The aim of this study was to evaluate the amount of apically extruded debris and irrigant during cleaning and shaping with different file systems. Seventy-five extracted mandibular premolar and canine teeth were randomly divided into five groups (n = 15). The root canals were instrumented by using Mtwo (VDW, Munich, Germany), Twisted File (TF; Sybron Endo, Orange, California, USA), Reciproc (VDW), Self-Adjusting File (SAF; Re Dent-Nova, Ra'anana, Israel) systems, and K-type stainless steel hand file (Mani Inc., Tochigi, Japan). Apically extruded debris was collected and dried in preweighed Eppendorf tubes. Extruded irrigant was collected from the cannula by using a plastic insulin syringe. After drying, the amount of apically extruded debris was established by subtracting the initial weight from the final weight of the tubes. After performing the Kolmogorov-Smirnov test, data were analyzed by Kruskall-Wallis test at a significance level of P value less than 0.05. There were no statistically significant differences between the file systems in terms of debris and irrigant extrusion. Mtwo group produced the highest mean debris extrusion value and the TF group produced the less compared with the other files. The least mean irrigant extrusion value was associated with the SAF group, whereas the highest mean irrigant extrusion value was associated with the Reciproc group. Under the conditions of the study, all file systems were associated with apical debris and irrigant extrusion.

  4. Absorbable Suture as an Apical Matrix in Single Visit Apexification with Mineral Trioxide Aggregate

    Directory of Open Access Journals (Sweden)

    Ayush Goyal

    2016-01-01

    Full Text Available Several procedures have been recommended to induce the root end barrier formation in teeth with open apices. Conventional treatment for such cases will require many appointments with an average duration of 12.9 months. During this period, the root canal is susceptible to reinfection from around the provisional restoration, which may promote apical periodontitis and arrest of apical repair. Mineral trioxide aggregate (MTA has been successfully used for one visit apexification wherein the root canal can be obturated within 24 hours after placement of MTA. Using a matrix prior to the placement of MTA avoids its extrusion, reduces leakage in the sealing material, and allows favorable response of the periapical tissues. This report presents a case of apexification where an absorbable suture was used as an apical matrix. Use of an absorbable suture circumvents all the problems associated with other conventional materials. Conclusion. Placement of the matrix made from the suture material is predictable and is easily positioned at the apex and the length can be adjusted as required. 10-month follow-up of the case shows resorbed matrix and bone healing in the periapical region. The patient was asymptomatic during the whole follow-up period and tooth exhibited mobility within physiologic limits and was functioning normally.

  5. Single Versus Multi-visit Endodontic Treatment of Teeth with Apical ...

    African Journals Online (AJOL)

    [3] Treatment of AP aims at the removal of cause, i.e., bacteria present in the canal. Single Versus Multi-visit Endodontic Treatment of. Teeth with Apical Periodontitis: An In vivo ... periapical healing of teeth with AP treated in (a) single visit versus (b) two visits, either with or .... The canal was left empty, and access cavity was.

  6. Comparative evaluation of total RNA extraction methods in Theobroma cacao using shoot apical meristems.

    Science.gov (United States)

    Silva, D V; Branco, S M J; Holanda, I S A; Royaert, S; Motamayor, J C; Marelli, J P; Corrêa, R X

    2016-03-04

    Theobroma cacao is a species of great economic importance with its beans used for chocolate production. The tree has been a target of various molecular studies. It contains many polyphenols, which complicate the extraction of nucleic acids with the extraction protocols requiring a large amount of plant material. These issues, therefore, necessitate the optimization of the protocols. The aim of the present study was to evaluate different methods for extraction of total RNA from shoot apical meristems of T. cacao 'CCN 51' and to assess the influence of storage conditions for the meristems on the extraction. The study also aimed to identify the most efficient protocol for RNA extraction using a small amount of plant material. Four different protocols were evaluated for RNA extraction using one shoot apical meristem per sample. Among these protocols, one that was more efficient was then tested to extract RNA using four different numbers of shoot apical meristems, subjected to three different storage conditions. The best protocol was tested for cDNA amplification using reverse transcription-polymerase chain reaction; the cDNA quality was determined to be satisfactory for molecular analyses. The study revealed that with the best RNA extraction protocol, one shoot apical meristem was sufficient for extraction of high-quality total RNA. The results obtained might enable advances in genetic analyses and molecular studies using reduced amount of plant material.

  7. Transmission electron microscopy of apical cells of Sphacelaria spp. (Sphacelariales, Phaeophyceae)

    NARCIS (Netherlands)

    Prud’homme van Reine, W.F.; Star, W.

    1981-01-01

    The ultrastructure of apical cells of six species of Sphacelaria (S. arctica, S. cirrosa, S. nana, S. racemosa, S. radicans, and S. rigidula) is studied here. In most details such as ultrastructure of chloroplasts, mitochondria, microbodies, nuclei and centrioles all Sphacelaria species studied are

  8. Single Versus Multi‑visit Endodontic Treatment of Teeth with Apical ...

    African Journals Online (AJOL)

    Background: Apical periodontitis (AP) is one of the most prevalent diseases of the teeth. Treatment of AP is based on the removal of the cause, i.e., bacteria from the root canals. Achievement of adequate bacterial eradication in one appointment treatment remains a controversy. Aim: This prospective study was conducted ...

  9. [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. © Georg Thieme Verlag KG Stuttgart · New York.

  10. The preparation of Dermatophilus congolensis antigen and its testing by means of immunodiffusion test and electrophoresis

    Directory of Open Access Journals (Sweden)

    Djaenudin Gholib

    1998-10-01

    Full Text Available The filtrate antigen ofDermatophilus congolensis was prepared based on the Makinde method, whereas the whole cell antigen was based on the Bida and Kelley method. Filtrate antigen of Dermatophilus congolensis has been tested with positive serum from experimental animals, guinea pigs and sheep by means of immurrodiffitsion test and electrophoresis . Positive serum was produced by inoculation of whole cell antigen of D. congolensis to the animals . The results showed that the immunodiffusion test resulted in one and two precipitation lines with positive serum of sheep and guinea pigs respectively. Electrophoresis SDS-PAGE presented about 8 bands with molecular weight in the range from above 30 kD to more than 94 kD. The bands were then transferred into nitrocellulose membrane and gave positive reaction with positive serum from sheep.

  11. Molecular typing of human platelet and neutrophil antigens (HPA and HNA).

    Science.gov (United States)

    Veldhuisen, Barbera; Porcelijn, Leendert; Ellen van der Schoot, C; de Haas, Masja

    2014-04-01

    Genotyping is an important tool in the diagnosis of disorders involving allo-immunisation to antigens present on the membranes of platelets and neutrophils. To date 28 human platelet antigens (HPAs) have been indentified on six polymorphic glycoproteins on the surface of platelets. Antibodies against HPAs play a role in foetal and neonatal alloimmune thrombocytopenia (FNAIT), post-transfusion purpura (PTP) and refractoriness to donor platelets. The 11 human neutrophil antigens (HNAs) described to date have been indentified on five polymorphic proteins on the surface of granulocytes. Antibodies to HNAs are implicated with foetal and neonatal alloimmune neutropenia (FNAIN), autoimmune neutropenia (AIN) and transfusion related acute lung injury (TRALI). In this report, we will review the molecular basis and techniques currently available for the genotyping of human platelet and neutrophil antigens. Copyright © 2014 Elsevier Ltd. All rights reserved.

  12. Elucidating the mechanisms of protein antigen adsorption to the CAF/NAF liposomal vaccine adjuvant systems

    DEFF Research Database (Denmark)

    Hamborg, Mette; Rose, Fabrice; Jorgensen, Lene

    2014-01-01

    is generally known about how antigens and adjuvants interact at the molecular level. The aim of this study was to elucidate the mechanisms of interactions between the equally sized, but oppositely charged model protein antigens α-lactalbumin and lysozyme, and i) the clinically tested cationic liposomal......The reverse vaccinology approach has recently resulted in the identification of promising protein antigens, which in combination with appropriate adjuvants can stimulate customized, protective immune responses. Although antigen adsorption to adjuvants influences vaccine efficacy and safety, little...... interaction with the zwitterionic liposomes. In contrast, the net cationic lysozyme showed very little interaction with either types of liposome. Adsorption of α-lactalbumin altered its tertiary structure, affected lipid membrane packing below and above the phase transition temperature, and neutralized...

  13. Effects of proteolytic enzymes and neuraminidase on the I and i erythrocyte antigen sites

    International Nuclear Information System (INIS)

    Doinel, C.; Ropars, C.; Salmon, C.

    1978-01-01

    Homogeneous cold agglutinins, purified and labelled with 125 I, have been used in a study of the effects of neuraminidase and proteolytic enzymes on the I and i reactivities of human adult erythrocytes. Measurements were made of antigen site numbers, equilibrium constants and thermodynamic parameters. There was enhanced reactivity after enzyme treatment as well as after the release of N-acetylneuraminic acid. Steric factors were shown to be of primary importance in the accessibility of the I and i antigenic determinant. After enzyme treatment, the antigenic structures became more homogeneous in their reaction with antibodies. The heterogeneity of binding constants observed with antigenic determinants of non-treated erythrocytes is probably due to the wide range of spatial distribution of these receptors within the membrane. (author)

  14. Apical wiring technique in surgical treatment of adolescent idiopathic scoliosis: the intermediate outcomes between Lenke types.

    Science.gov (United States)

    Chaiyamongkol, Weera; Klineberg, Eric O; Gupta, Munish C

    2013-02-01

    Retrospective cohort study of apical wiring technique for surgical treatment of adolescent idiopathic scoliosis patients. To evaluate the intermediate outcome of adolescent idiopathic scoliosis patients treated with apical wiring technique. Regarding surgical treatment for adolescent idiopathic scoliosis, there are reports of the superiority of coronal plane correction using all-screw constructs compared with all-hook or hybrid constructs. Major limitations of all-screw constructs are thoracic hypokyphosis and increased proximal junctional kyphosis. There are few reports about apical wiring technique outcomes and no reports of this surgical technique comparing Lenke types. Consecutive adolescent idiopathic scoliosis patients treated with apical wiring technique were analyzed. Inclusion criteria were patient 21 years old or younger, who underwent a posterior only surgical correction with a follow-up of at least 2 years. Radiographic data were analyzed according to the criteria given by Lenke and colleagues. The radiographic parameters included global balance, Cobb angle, curve flexibility, apical vertebral translation, tilt angle of lower instrumented vertebra, angle of caudal disk to the lower instrumented vertebra, thoracic kyphosis, lumbar lordosis, and proximal junctional angle preoperatively and postoperatively. There was a significant improvement of Cobb angle in every Lenke type with an average correction of 74.6%. At latest follow-up, the average loss of correction was 3.5% (1.9 degrees). The greatest loss of correction for the minor curve of Lenke 2 was 14.1% (5.8 degrees). There was an improvement of thoracic kyphosis in hypokyphosis group (1.9-21.1 degrees; Pwiring technique for surgical treatment of adolescent idiopathic scoliosis provides an average correction of 71.1% at 3.4 years of average follow-up with harmonious sagittal plane correction even with preexisting abnormal thoracic kyphosis.

  15. Wizard CD Plus and ProTaper Universal: analysis of apical transportation using new software.

    Science.gov (United States)

    Giannastasio, Daiana; Rosa, Ricardo Abreu da; Peres, Bernardo Urbanetto; Barreto, Mirela Sangoi; Dotto, Gustavo Nogara; Kuga, Milton Carlos; Pereira, Jefferson Ricardo; Só, Marcus Vinícius Reis

    2013-01-01

    This study has two aims: 1) to evaluate the apical transportation of the Wizard CD Plus and ProTaper Universal after preparation of simulated root canals; 2) to compare, with Adobe Photoshop, the ability of a new software (Regeemy) in superposing and subtracting images. Twenty five simulated root canals in acrylic-resin blocks (with 20º curvature) underwent cone beam computed tomography before and after preparation with the rotary systems (70 kVp, 4 mA, 10 s and with the 8×8 cm FoV selection). Canals were prepared up to F2 (ProTaper) and 24.04 (Wizard CD Plus) instruments and the working length was established to 15 mm. The tomographic images were imported into iCAT Vision software and CorelDraw for standardization. The superposition of pre- and post-instrumentation images from both systems was performed using Regeemy and Adobe Photoshop. The apical transportation was measured in millimetres using Image J. Five acrylic resin blocks were used to validate the superposition achieved by the software. Student's t-test for independent samples was used to evaluate the apical transportation achieved by the rotary systems using each software individually. Student's t-test for paired samples was used to compare the ability of each software in superposing and subtracting images from one rotary system per time. The values obtained with Regeemy and Adobe Photoshop were similar to rotary systems (P>0.05). ProTaper Universal and Wizard CD Plus promoted similar apical transportation regardless of the software used for image's superposition and subtraction (P>0.05). Wizard CD Plus and ProTaper Universal promoted little apical transportation. Regeemy consists in a feasible software to superpose and subtract images and appears to be an alternative to Adobe Photoshop.

  16. Wizard CD Plus and ProTaper Universal: analysis of apical transportation using new software

    Directory of Open Access Journals (Sweden)

    Daiana Giannastasio

    2013-09-01

    Full Text Available OBJECTIVE: This study has two aims: 1 to evaluate the apical transportation of the Wizard CD Plus and ProTaper Universal after preparation of simulated root canals; 2 to compare, with Adobe Photoshop, the ability of a new software (Regeemy in superposing and subtracting images. MATERIAL AND METHODS: Twenty five simulated root canals in acrylic-resin blocks (with 20º curvature underwent cone beam computed tomography before and after preparation with the rotary systems (70 kVp, 4 mA, 10 s and with the 8×8 cm FoV selection. Canals were prepared up to F2 (ProTaper and 24.04 (Wizard CD Plus instruments and the working length was established to 15 mm. The tomographic images were imported into iCAT Vision software and CorelDraw for standardization. The superposition of pre- and post-instrumentation images from both systems was performed using Regeemy and Adobe Photoshop. The apical transportation was measured in millimetres using Image J. Five acrylic resin blocks were used to validate the superposition achieved by the software. Student's t-test for independent samples was used to evaluate the apical transportation achieved by the rotary systems using each software individually. Student's t-test for paired samples was used to compare the ability of each software in superposing and subtracting images from one rotary system per time. RESULTS: The values obtained with Regeemy and Adobe Photoshop were similar to rotary systems (P>0.05. ProTaper Universal and Wizard CD Plus promoted similar apical transportation regardless of the software used for image's superposition and subtraction (P>0.05. CONCLUSION: Wizard CD Plus and ProTaper Universal promoted little apical transportation. Regeemy consists in a feasible software to superpose and subtract images and appears to be an alternative to Adobe Photoshop.

  17. 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.

  18. Membrane organization determines barrier properties of endothelial cells and short-chain sphingolipid-facilitated doxorubicin influx.

    Science.gov (United States)

    van Hell, A J; Klymchenko, A; Gueth, D M; van Blitterswijk, W J; Koning, G A; Verheij, M

    2014-09-01

    The endothelial lining and its outer lipid membrane are the first major barriers drug molecules encounter upon intravenous administration. Our previous work identified lipid analogs that counteract plasma membrane barrier function for a series of amphiphilic drugs. For example, short-chain sphingolipids (SCS), like N-octanoyl-glucosylceramide, effectively elevated doxorubicin accumulation in tumor cells, both in vitro and in vivo, and in endothelial cells, whereas other (normal) cells remained unaffected. We hypothesize here that local membrane lipid composition and the degree of lipid ordering define SCS efficacy in individual cells. To this end, we study the differential effect of SCS on bovine aortic endothelial cells (BAEC) in its confluent versus proliferative state, as a model system. While their (plasma membrane) lipidome stays remarkably unaltered when BAECs reach confluency, their lipids segregate to form apical and basolateral domains. Using probe NR12S, we reveal that lipids in the apical membrane are more condensed/liquid-ordered. SCS preferentially attenuate the barrier posed by these condensed membranes and facilitate doxorubicin influx in these particular membrane regions. We confirm these findings in MDCK cells and artificial membranes. In conclusion, SCS-facilitated drug traversal acts on condensed membrane domains, elicited by confluency in resting endothelium. Copyright © 2014 Elsevier B.V. All rights reserved.

  19. Removal of GPI-anchored membrane proteins causes clustering of lipid microdomains in the apical head area of porcine sperm

    NARCIS (Netherlands)

    Boerke, Arjan; van der Lit, Joost; Lolicato, Francesca; Stout, Tom A E; Helms, J Bernd; Gadella, Bart M

    2014-01-01

    The release of extracellular proteins is a part of the sperm capacitation process; this allows the sperm surface reorganization that enables the sperm to fertilize an oocyte. Some of the components released are 'decapacitation factors', an uncoordinated or early release of which may cause

  20. The breast cancer resistance protein transporter ABCG2 is expressed in the human kidney proximal tubule apical membrane.

    NARCIS (Netherlands)

    Huls, M.; Brown, C.D.; Windass, A.S.; Sayer, R.; Heuvel, J.J.M.W. van den; Heemskerk, S.; Russel, F.G.M.; Masereeuw, R.

    2008-01-01

    The Breast Cancer Resistance Protein (BCRP/ABCG2) is a transporter restricting absorption and enhancing excretion of many compounds including anticancer drugs. This transporter is highly expressed in many tissues; however, in human kidney, only the mRNA was found in contrast to the mouse kidney,

  1. 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

  2. Associative link of clinical manifestations of the secondary syphilis of skin and mucosa with histocompatibility antigens Class I

    Directory of Open Access Journals (Sweden)

    S. V. Koshkin

    2017-01-01

    Full Text Available Sixty patients with different clinical symptoms of secondary syphilis (ulcer chancres, pustular syphilis, hypertrophic papules, widespread leukoderma and alopecia were examined in order to study the distribution pattern of histocompatibility antigens of the first class in patients with secondary syphilis of the skin and mucous membranes. As a result of the study, the presence of an associative relationship between the distribution pattern of histocompatibility antigens of the first class and various clinical manifestations in patients with secondary syphilis was established.

  3. Expression and distribution of leptospiral outer membrane components during renal infection of hamsters

    NARCIS (Netherlands)

    Barnett, J. K.; Barnett, D.; Bolin, C. A.; Summers, T. A.; Wagar, E. A.; Cheville, N. F.; Hartskeerl, R. A.; Haake, D. A.

    1999-01-01

    The outer membrane of pathogenic Leptospira species grown in culture media contains lipopolysaccharide (LPS), a porin (OmpL1), and several lipoproteins, including LipL36 and LipL41. The purpose of this study was to characterize the expression and distribution of these outer membrane antigens during

  4. 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

  5. From biological membranes to biomimetic model membranes

    Directory of Open Access Journals (Sweden)

    Eeman, M.

    2010-01-01

    Full Text Available Biological membranes play an essential role in the cellular protection as well as in the control and the transport of nutrients. Many mechanisms such as molecular recognition, enzymatic catalysis, cellular adhesion and membrane fusion take place into the biological membranes. In 1972, Singer et al. provided a membrane model, called fluid mosaic model, in which each leaflet of the bilayer is formed by a homogeneous environment of lipids in a fluid state including globular assembling of proteins and glycoproteins. Since its conception in 1972, many developments were brought to this model in terms of composition and molecular organization. The main development of the fluid mosaic model was made by Simons et al. (1997 and Brown et al. (1997 who suggested that membrane lipids are organized into lateral microdomains (or lipid rafts with a specific composition and a molecular dynamic that are different to the composition and the dynamic of the surrounding liquid crystalline phase. The discovery of a phase separation in the plane of the membrane has induced an explosion in the research efforts related to the biology of cell membranes but also in the development of new technologies for the study of these biological systems. Due to the high complexity of biological membranes and in order to investigate the biological processes that occur on the membrane surface or within the membrane lipid bilayer, a large number of studies are performed using biomimicking model membranes. This paper aims at revisiting the fundamental properties of biological membranes in terms of membrane composition, membrane dynamic and molecular organization, as well as at describing the most common biomimicking models that are frequently used for investigating biological processes such as membrane fusion, membrane trafficking, pore formation as well as membrane interactions at a molecular level.

  6. 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.

  7. Elevated cAMP increases aquaporin-3 plasma membrane diffusion

    DEFF Research Database (Denmark)

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

    2014-01-01

    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...

  8. Apical extrusion of sodium hypochlorite using different root canal irrigation systems.

    Science.gov (United States)

    Mitchell, Ross P; Baumgartner, J Craig; Sedgley, Christine M

    2011-12-01

    Root canal irrigation carries a risk of extrusion of irrigant into the periapical tissues. The objective of this study was to compare different irrigation systems in matched pairs of teeth prepared to an apical size of 35.06 and 50.06 by measuring the frequency and extent of apical extrusion of sodium hypochlorite (NaOCl) into a simulated periapical environment. The null hypothesis was tested that there is no difference between systems. Bilaterally matched pairs (n = 10) of single-canal extracted human anterior teeth were instrumented to an apical size of either 35.06 or 50.06. Teeth were embedded in a gel containing the pH-sensitive dye M-cresol purple that changes from yellow at pH 7.4 to purple at pH 9. Root canals were irrigated with 6% NaOCl (pH 11) by using EndoActivator (EA), EndoVac (EV), Rispi-Sonic/MicroMega 1500 (MM), passive ultrasonic irrigation (PUI), and syringe irrigation with a slot-tipped needle (SN), so that each tooth underwent all irrigation procedures in a randomized crossover design. Apical extrusion was evaluated by image analyses. The frequency of extrusion was less in teeth with apical preparation size 35.06 (36%) compared with 50.06 (60%) (P = .014) and was dependent on the irrigation system in 35.06 (P = .039) but not 50.06 groups. In the 35.06 group the frequency of extrusion was less for EV than for MM and SN (both P = .029). The extent of extrusion was less for MM compared with PUI (P = .024) and SN (P = .046) in the 35.06 group and greater for SN compared with all other systems in the 50.06 group (P root canal irrigation system and apical preparation size. The extent of extrusion depended on the irrigation system, with syringe and slotted-needle irrigation resulting in the greatest extent of extrusion. Copyright © 2011 American Association of Endodontists. Published by Elsevier Inc. All rights reserved.

  9. [Clinical characteristics and speech therapy of lingua-apical articulation disorder].

    Science.gov (United States)

    Zhang, Feng-hua; Jin, Xing-ming; Zhang, Yi-wen; Wu, Hong; Jiang, Fan; Shen, Xiao-ming

    2006-03-01

    To explore the clinical characteristics and speech therapy of 62 children with lingua-apical articulation disorder. Peabody Picture Vocabulary Test (PPVT), Gesell development scales (Gesell), Wechsler Intelligence Scale for Preschool Children (WPPSI) and speech test were performed for 62 children at the ages of 3 to 8 years with lingua-apical articulation disorder. PPVT was used to measure receptive vocabulary skills. GESELL and WPPSI were utilized to represent cognitive and non-verbal ability. The speech test was adopted to assess the speech development. The children received speech therapy and auxiliary oral-motor functional training once or twice a week. Firstly the target sound was identified according to the speech development milestone, then the method of speech localization was used to clarify the correct articulation placement and manner. It was needed to change food character and administer oral-motor functional training for children with oral motor dysfunction. The 62 cases with the apical articulation disorder were classified into four groups. The combined pattern of the articulation disorder was the most common (40 cases, 64.5%), the next was apico-dental disorder (15 cases, 24.2%). The third was palatal disorder (4 cases, 6.5%) and the last one was the linguo-alveolar disorder (3 cases, 4.8%). The substitution errors of velar were the most common (95.2%), the next was omission errors (30.6%) and the last was absence of aspiration (12.9%). Oral motor dysfunction was found in some children with problems such as disordered joint movement of tongue and head, unstable jaw, weak tongue strength and poor coordination of tongue movement. Some children had feeding problems such as preference of eating soft food, keeping food in mouths, eating slowly, and poor chewing. After 5 to 18 times of therapy, the effective rate of speech therapy reached 82.3%. The lingua-apical articulation disorders can be classified into four groups. The combined pattern of the

  10. Deviations of Mesial Root Canals of Mandibular First Molar Teeth at the Apical Third: A Micro-computed Tomographic Study.

    Science.gov (United States)

    Keles, Ali; Keskin, Cangül

    2018-04-19

    The present study aimed to quantitatively analyze apical foramen deviations of mesial root canals of mandibular first molar teeth by means of micro-computed tomographic (micro-CT) imaging. Micro-CT images of the mesial roots of 109 mandibular first molar teeth with independent mesiobuccal (MB) and mesiolingual (ML) root canals were analyzed. The deviations of the apical foramina of the MB, ML, and middle mesial root canals from the anatomic apex were measured. The vertical distance between the apical foramina of each mesial root canal in relation to each other was also calculated. The distances from the apical foramina of the MB, ML, and middle mesial root canals to the anatomic apex of the mesial root were up to 2.51 mm, 3.21 mm, and 5.67 mm, respectively. There was no significant difference between the deviations of MB and ML root canals from each other (P > .05). The middle mesial root canal showed the greatest deviation compared with the MB and ML canals (P < .05). The apical foramina of mesial root canals of mandibular first molar teeth showed greater variations from each other and anatomic apices than previously reported. Clinically, the use of electronic apex locators for the detection of minor apical foramen of each mesial root canal is of the utmost important. Copyright © 2018 American Association of Endodontists. Published by Elsevier Inc. All rights reserved.

  11. Adenovirus Entry From the Apical Surface of Polarized Epithelia Is Facilitated by the Host Innate Immune Response

    Science.gov (United States)

    Kotha, Poornima L. N.; Sharma, Priyanka; Kolawole, Abimbola O.; Yan, Ran; Alghamri, Mahmoud S.; Brockman, Trisha L.; Gomez-Cambronero, Julian; Excoffon, Katherine J. D. A.

    2015-01-01

    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. PMID:25768646

  12. Screening Donors for Rare Antigen Constellations.

    Science.gov (United States)

    Wagner, Franz F

    2009-01-01

    SCREENING BLOOD DONORS FOR RARE ANTIGEN CONSTELLATIONS HAS BEEN IMPLEMENTED USING SIMPLE PCR METHODS: PCR with enzyme digestion has been used to type donor cohorts for Dombrock antigens, and PCR with sequence-specific priming to identify donors negative for antigens of high frequency. The advantages and disadvantages of the methods as well as their current state is discussed.

  13. 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

    The Cl- -current through toad skin epithelium depends on the potential in a way consistent with a potential-controlled Cl- permeability. Computer analysis of the Koefoed-Johnsen Ussing two-membrane model provided with constant membrane permeabilities indicates that the voltage- and time-dependent......The Cl- -current through toad skin epithelium depends on the potential in a way consistent with a potential-controlled Cl- permeability. Computer analysis of the Koefoed-Johnsen Ussing two-membrane model provided with constant membrane permeabilities indicates that the voltage- and time...... resemble experimental observations. This extension of the classic frog skin model implies that the Cl- permeability is activated by a voltage change caused by the inward Na+ current through the apical membrane....

  14. Type I collagen gel induces Madin-Darby canine kidney cells to become fusiform in shape and lose apical-basal polarity.

    Science.gov (United States)

    Zuk, A; Matlin, K S; Hay, E D

    1989-03-01

    In the embryo, epithelia give rise to mesenchyme at specific times and places. Recently, it has been reported (Greenburg, G., and E. D. Hay. 1986. Dev. Biol. 115:363-379; Greenberg, G., and E. D. Hay. 1988. Development (Camb.). 102:605-622) that definitive epithelia can give rise to fibroblast-like cells when suspended within type I collagen gels. We wanted to know whether Madin-Darby canine kidney (MDCK) cells, an epithelial line, can form mesenchyme under similar conditions. Small explants of MDCK cells on basement membrane were suspended within or placed on top of extracellular matrix gels. MDCK cells on basement membrane gel are tall, columnar in shape, and ultrastructurally resemble epithelia transporting fluid and ions. MDCK explants cultured on type I collagen gel give rise to isolated fusiform-shaped cells that migrate over the gel surface. The fusiform cells extend pseudopodia and filopodia, lose cell membrane specializations, and develop an actin cortex around the entire cell. Unlike true mesenchymal cells, which express vimentin and type I collagen, fusiform cells produce both keratin and vimentin, continue to express laminin, and do not turn on type I collagen. Fusiform cells are not apically-basally polarized, but show mesenchymal cell polarity. Influenza hemagglutinin and virus budding localize to the front end or entire cell surface. Na,K-ATPase occurs intracellularly and also symmetrically distributes on the cell surface. Fodrin becomes diffusely distributed along the plasma membrane, ZO-1 cannot be detected, and desmoplakins distribute randomly in the cytoplasm. The loss of epithelial polarity and acquisition of mesenchymal cell polarity and shape by fusiform MDCK cells on type I collagen gel was previously unsuspected. The phenomenon may offer new opportunities for studying cytoplasmic and nuclear mechanisms regulating cell shape and polarity.

  15. Traffic pathways of Plasmodium vivax antigens during intraerythrocytic parasite development.

    Science.gov (United States)

    Bracho, Carmen; Dunia, Irene; De, La Rosa Mercedes; Benedetti, Ennio-Lucio; Perez, Hilda A

    2002-03-01

    We investigated the secretory traffic of a Plasmodium vivax antigen (Pv-148) synthesised by the parasite during the blood cycle, exported into the host cell cytosol and then transported to the surface membrane of the infected erythrocyte. Studies of the ultrastructure of erythrocytes infected with P. vivax showed that intracellular schizogony is accompanied by the generation of parasite-induced membrane profiles in the erythrocyte cytoplasm. These structures are detectable soon after the parasite invades the erythrocyte and develop an elaborate organisation, leading to a tubovesicular membrane (TVM) network, in erythrocytes infected with mature trophozoites. Interestingly, the clefts formed stacked, flattened cisternae resembling a classical Golgi apparatus. The TVM network stained with the fluorescent Golgi marker Bodipy-ceramide. Specific immunolabelling showed that Pv-148 was transferred from the parasite to the erythrocyte surface membrane via the clefts and the TVM network. These findings suggest that the TVM network is part of the secretory pathways involved in parasite protein transport across the Plasmodium-infected erythrocyte and that Pv- 148 may represent a marker that links the parasite with the host cell cytoplasm and, in turn, with the extracellular milieu.

  16. Function and Dynamics of Tetraspanins during Antigen Recognition and Immunological Synapse Formation

    Directory of Open Access Journals (Sweden)

    Vera eRocha-Perugini

    2016-01-01

    Full Text Available Tetraspanin-enriched microdomains (TEMs are specialized membrane platforms driven by protein-protein interactions that integrate membrane receptors and adhesion molecules. Tetraspanins participate in antigen recognition and presentation by antigen presenting cells (APCs through the organization of pattern recognition receptors (PRRs and their downstream induced-signaling, as well as the regulation of MHC-II-peptide trafficking. T lymphocyte activation is triggered upon specific recognition of antigens present on the APC surface during immunological synapse (IS formation. This dynamic process is characterized by a defined spatial organization involving the compartmentalization of receptors and adhesion molecules in specialized membrane domains that are connected to the underlying cytoskeleton and signaling molecules. Tetraspanins contribute to the spatial organization and maturation of the IS by controlling receptor clustering and local accumulation of adhesion receptors and integrins, their downstream signaling and linkage to the actin cytoskeleton. This review offers a perspective on the important role of TEMs in the regulation of antigen recognition and presentation, and in the dynamics of IS architectural organization.

  17. Facilitation of Endosomal Recycling by an IRG Protein Homolog Maintains Apical Tubule Structure in Caenorhabditis elegans

    Science.gov (United States)

    Grussendorf, Kelly A.; Trezza, Christopher J.; Salem, Alexander T.; Al-Hashimi, Hikmat; Mattingly, Brendan C.; Kampmeyer, Drew E.; Khan, Liakot A.; Hall, David H.; Göbel, Verena; Ackley, Brian D.; Buechner, Matthew

    2016-01-01

    Determination of luminal diameter is critical to the function of small single-celled tubes. A series of EXC proteins, including EXC-1, prevent swelling of the tubular excretory canals in Caenorhabditis elegans. In this study, cloning of exc-1 reveals it to encode a homolog of mammalian IRG proteins, which play roles in immune response and autophagy and are associated with Crohn’s disease. Mutants in exc-1 accumulate early endosomes, lack recycling endosomes, and exhibit abnormal apical cytoskeletal structure in regions of enlarged tubules. EXC-1 interacts genetically with two other EXC proteins that also affect endosomal trafficking. In yeast two-hybrid assays, wild-type and putative constitutively active EXC-1 binds to the LIM-domain protein EXC-9, whose homolog, cysteine-rich intestinal protein, is enriched in mammalian intestine. These results suggest a model for IRG function in forming and maintaining apical tubule structure via regulation of endosomal recycling. PMID:27334269

  18. 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

  19. One versus two appointment endodontic treatment on teeth with apical periodontitis

    Directory of Open Access Journals (Sweden)

    Juni Jekti Nugroho

    2016-06-01

    Full Text Available The microbiological purpose of endodontic treatment on tooth with apical periodontitis is reducing microbes to achieve periradicular tissue healing and prevent microbes recolonized. This is achieved by antimicrobial measures, including chemomechanical procedures and intracanal medication, with root canal obturation steps. One of the problem in endodontic is the planning consideration to treat with one or two visits. Does intra-appointment medication is an absolute measure to promote desinfection and treatment success. To provide a guideline about  one or two visit endodontic treatment decision making for dental professionals. Root canal system on tooth with apical periodontitis treated in two visit endodontic and calcium hydroxide as the intra-appointment medication achieved better microbiological status than the one treated in one visit endodontic.

  20. Inadvertent Apical Extrusion of Sodium Hypochlorite with Evaluation by Dental Volumetric Tomography

    Science.gov (United States)

    Başer Can, Elif Delve; Karapınar Kazandağ, Meriç; Kaptan, Rabia Figen

    2015-01-01

    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. PMID:25883812