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Cha o 1 gene organization.
No full textThe genomic DNA of the Cha o 1 gene from the plus tree Miho 6 was amplified and sequenced (Genbank accession number #LC586088). (A) Comparison with the cDNA sequence of Cha o 1 reported previously (Genbank accession number #D45404) [14]. The exon and intron structures were determined. The DNA sequence for the exon of Cha o 1 from Miho 6 was identical to reported cDNA and was used as the reference sequence for NGS in this report. Cha o 1 genomic DNA of the Shisui 1 plus tree has a second intron that is ~0.6 kb longer and the 1810th nucleotide (asterisk) is mutated from cytosine to adenine, which gives rise to the L262I substitution. The newly formed SNP afforded the PsiI restriction site, TTATAA. Primer pairs for PCR amplification and detection of the SNP are shown as arrows. (B) Electrophoresis of the restricted PCR product by PsiI. PCR products of Kinada 4, Miho 6 were not restricted by PsiI, which indicates that residue 262 of them are Leucine. The PCR product from Shisui 1 was completely restricted, which indicates that residue 262 is Isoleucine. From a mixture of Kinada 4 and Shisui 1, an artificial heterozygote for the substitution was prepared and its PCR product was partially restricted. A similar band pattern was obtained from Tanzawa 8, which indicates that residue 262 was a mixture of Leu and Ile. (C) PCR-RFLP analysis of the 47 mother trees from the seed orchards of Chiba Prefecture. PCR products of the 3rd exon of Cha o 1 were analyzed for the presence of the PsiI recognition site. Homozygotes (*) and heterozygotes (**) for the substitution of Cha o 1[L262I] are shown. N: negative control, PCR product without template treated with PsiI.</p
Cha o 3, a cypress pollen allergen, does not activate basophils in Japanese cypress pollinosis
No full textBackground: In Japan, pollinosis caused by the Japanese cypress (JCy) Chamaecyparis obtusa is among the very common seasonal allergies. In JCy pollinosis, Cha o 1 is the first major allergen, and Cha o 2 is the second major allergen. Recently, Cha o 3 was identified as a new JCy pollen allergen in JCy pollinosis. However, the relative contribution of Cha o 3 to JCy pollinosis compared with that of Cha o 1 and that of Cha o 2 has not been fully elucidated. Objective: This study aimed to clarify the allergenicity of Cha o 3 compared with that of Cha o 1 and Cha o 2 in JCy pollinosis. Methods: We recruited 27 patients with JCy pollinosis and performed the basophil activation test (BAT) with native (n) Cha o 1, Cha o 2, and Cha o 3 purified from JCy pollen. In addition, we a performed JCy-specific IgE suppression test. Results: In the BAT, 26 of 27 patients (96%) and 18 of 27 patients (67%) showed positive basophil activation in response to n Cha o 1 and n Cha o 2, respectively, as judged by CD203c expression. Little CD203c expression in response to n Cha o 3 was seen. The presence of n Cha o 3 marginally reduced the titer levels of JCy-specific IgE. Conclusion: Cha o 3 showed little ability to activate basophils and suppress JCy-specific IgE titers compared with Cha o 1 or Cha o 2 in patients with JCy pollinosis. Thus, Cha o 3 may not be a major allergen in JCy pollinosis
Comparison of the Cha o 2 and Cry j 2 amino acid sequences.
No full textPolymorphic residues in Cha o 2 (outlined letters), substituted residues (lower letters in the Cha o 2 sequence), IgE epitopes (boxed sequence), and the mouse mAb 9E7 epitope (grey shaded sequence) are shown.</p
Identification of Cha o 3 homolog Cry j 4 from Cryptomeria japonica (Japanese cedar) pollen: Limitation of the present Japanese cedar–specific ASIT
No full textBackground: About one-third of the Japanese population suffers from Japanese cedar pollinosis, which is frequently accompanied by Japanese cypress pollinosis. Recently, a novel major Japanese cypress pollen allergen, Cha o 3, was discovered. However, whether a Cha o 3 homolog is present in Japanese cedar pollen remains to be determined. Methods: Western blot analysis was performed using Cha o 3–specific antiserum. In addition, cloning of the gene encoding Cry j 4 was conducted using total cDNA from the male flower of Japanese cedar trees. Allergen potency and cross-reactivity were investigated using a T-cell proliferation assay, basophil activation test, and ImmunoCAP inhibition assay. Results: A low amount of Cha o 3 homolog protein was detected in Japanese cedar pollen extract. The deduced amino acid sequence of Cry j 4 showed 84% identity to that of Cha o 3. Cross-reactivity between Cry j 4 and Cha o 3 was observed at the T cell and IgE levels. Conclusions: Cry j 4 was discovered as a counterpart allergen of Cha o 3 in Japanese cedar pollen, with a relationship similar to that between Cry j 1–Cha o 1 and Cry j 2–Cha o 2. Our findings also suggest that allergen-specific immunotherapy (ASIT) using Japanese cedar pollen extract does not induce adequate immune tolerance to Cha o 3 due to the low amount of Cry j 4 in Japanese cedar pollen. Therefore, ASIT using Cha o 3 or cypress pollen extract coupled with Japanese cedar pollen extract is required in order to optimally control allergy symptoms during Japanese cypress pollen season. Keywords: Allergen, Cedar pollen, Cha o 3, Cry j 4, Japanese cedar pollinosi
Going Beyond Counting First Authors in Author Co-citation Analysis
Get PDFThe present study examines one of the fundamental aspects of author co-citation analysis (ACA) - the way co-citation
counts are defined. Co-citation counting provides the data on which all subsequent statistical analyses and mappings
are based, and we compare ACA results based on two different types of co-citation counting - the traditional type that
only counts the first one among a cited work's authors on the one hand and a non-traditional type that takes into
account the first 5 authors of a cited work on the other hand. Results indicate that the picture produced through this non-traditional author co-citation counting contains more coherent author groups and is therefore considerably clearer. However, this picture represents fewer specialties in the research field being studied than that produced through the traditional first-author co-citation counting when the same number of top-ranked authors is selected and analyzed. Reasons for these effects are discussed
Cross-reactivity between major IgE core epitopes on Cry j 2 allergen of Japanese cedar pollen and relevant sequences on Cha o 2 allergen of Japanese cypress pollen
No full textBackground: Cry j 2 and Cha o 2 are major allergens in Japanese cedar (Cryptomeria japonica; CJ) and Japanese cypress (Chamaecyparis obtusa; CO) pollen, respectively. Here, we assessed the epitopes related to the cross-reactivity between Cry j 2 and Cha o 2 using in vitro analyses.
Methods: Peptides were synthesized based on Cry j 2 sequential epitopes and relevant Cha o 2 amino acid sequences. Four representative monoclonal antibodies (mAbs) against Cry j 2 were used according to their epitope recognitions. Serum samples were collected from 31 patients with CJ pollinosis. To investigate cross-reactivity between Cry j 2 and Cha o 2, ELISA and inhibition ELISA were performed with mAbs and sera from patients with CJ pollinosis.
Results: Two of four mAbs had reactivity to both Cry j 2 and Cha o 2. Of these two mAbs, one mAb (T27) recognized the amino acid sequence 169KVVNGRTV176 on Cha o 2. This is related to the core epitope 169KWVNGREI176 on Cry j 2, which is an important IgE epitope. In addition, we found that these correlative sequences and purified allergens showed cross-reactivity between Cry j 2 and Cha o 2 in IgE of CJ patients.
Conclusions: We demonstrated the importance of 169KVVNGRTV176 in Cha o 2 for cross-reactivity with the Cry j 2 epitope 169KWVNGREI176, which plays an important role in allergenicity in CJ pollinosis. Our results are useful for the development of safer and more efficient therapeutic strategies for the treatment of CJ and CO pollen allergies
Comparison of amino acid sequences of isoallergen Cha o 2.0101 and those deduced from genomic sequence data of the Cha o 2 gene.
No full textThe N-terminal amino acid of isoallergen Cha o 2.0101 after processing is boxed and numbered as 1 [15, 16]. The three residues that differed in BAF32143 [22] are indicated and other residues are shown as ‘.’. Undetermined residues are shown as ‘–‘. Substitutions discovered in this study are shown below the BAF32143 sequence.</p
Variations on the Author
Get PDF“Variations on the Author” discusses two of Eduardo Coutinho’s recent films (Um Dia na Vida, from 2010, and Últimas Conversas, posthumously released in 2015) and their contribution to the general question of documentary authorship. The director’s filmography is characterized by a consistent yet self-effacing form of authorial self-inscription: Coutinho often features as an interviewer that rather than express opinions propels discourses; an interviewer that is good at listening. This mode of self-inscription characterizes him as an author who is not expressive but who is nonetheless markedly present on the screen. In Um Dia na Vida, however, Coutinho is completely absent form the image, while Últimas Conversas, on the contrary, includes a confessional prologue that moves the director from the margins to the center of his films. This article examines the ways in which these works stand out in the filmography of a director who offers new insights into the notion of cinematic authorship
Appropriate Similarity Measures for Author Cocitation Analysis
Get PDFWe provide a number of new insights into the methodological discussion about author cocitation analysis. We first argue that the use of the Pearson correlation for measuring the similarity between authors’ cocitation profiles is not very satisfactory. We then discuss what kind of similarity measures may be used as an alternative to the Pearson correlation. We consider three similarity measures in particular. One is the well-known cosine. The other two similarity measures have not been used before in the bibliometric literature. Finally, we show by means of an example that our findings have a high practical relevance.information science;Pearson correlation;cosine;similarity measure;author cocitation analysis
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