Life story narrative analysis of female agoraphobia

This dissertation presents theoretical and empirical research on the agoraphobia. The theoretical part outlines the history and diagnosis of agoraphobia since the mid- nineteenth century to the present through selected theories of this disorder. Concept of normality and diagnosis of agoraphobia disorder according to ICD -10, DSM IV and DSM V are also addressed. Space is also devoted to the philosophical concept of anxiety and body in relation to space. In the empirical part of the thesis the life story narrative analysis as well as analysis of narrative interviews with women suffering from agoraphobia was carried out. Ten women represent the sample. Received research data were analysed using the categorial- formal analysis. Following research objectives were set: to analyse importance of the disease in lives of participants, to describe the individual specific form of the disease, to analyse individual meaning and purpose attributed by the participants to their disease, to describe diverse aspects of life depicting. The aim was also to learn about the ways participants manage their disease including the use of therapy. Life story narrative analysis of female agoraphobia proved to be a suitable method for understanding the very meaning of agoraphobia disorder and also the meaning that is being..

Alignment of predicted amino acid sequences from entire cruzipain of <i>T. cruzi</i> (TcI, TcII, TcIII, TcVI and Tcbat) and homologues from <i>T. cruzi</i>-like (<i>T. c. marinkellei</i> and <i>T. dionisii</i>), <i>T. rangeli</i> and <i>T. b. brucei</i>.

<p>Pre, pro, catalytic domain and C-terminal extension amino acid sequences of cruzipain genes from <i>T. cruzi</i> Sylvio X10.6 and G (TcI), TCC1994 (Tcbat), Y and Esmeraldo cl3 (TcII), M6241 cl6 (TcIII), CL Brener (TcVI) Non-Esmeraldo-like (TcIII) and Esmeraldo-like (TcII) haplotypes and homologues from <i>T. c. marinkellei</i> (344), <i>T. dionisii</i> (211), <i>T. rangeli</i> (LDG and AM80) and <i>T. b. brucei</i> (TREU 927). The CATL family signatures of pro-domain motifs ERFININ (ERFN) and GNFD (GTFD) are indicated in bold and underlined, the subsites S1, S2 and S2′ are in bold, and the conserved Trp181 are indicated by (*).The glutamine [Q] of the oxyanion hole, cysteine [C], histidine [H] and asparagine [N] of catalytic triad in the catalytic domain, and 8 cysteines in the C-terminal extension are indicated by arrow heads.</p

Polymorphism and network analyses of catalytic domain sequences of cruzipain genes from <i>T. cruzi</i> isolates of TcI-VI and Tcbat.

<p>(A) Polymorphic nucleotide sites on catalytic domains of cruzipain encoding genes; (B) Network based on polymorphic nucleotides constructed with the Neighbour-Net algorithm excluding all conserved sites and with Uncorrected p-distance. The numbers in nodes correspond to bootstrap values from 100 replicates. CLBrener1-5 are sequences from TriTrypDB: Tc00.1047053509429.320, Tc00.1047053507537.20, Tc00.1047053507603.270, Tc00.1047053507603.260 and Tc00.1047053507537.10; *GenBank accession numbers of all sequences included in these analyses are listed on Table1. Major types of sequences from <i>T. cruzi</i> isolates of different DTUs are indicated by different colors according to the legend.</p

Synteny of a locus containing cruzipain genes in <i>T. cruzi</i>, <i>T. dionisii</i> and <i>T. b. brucei</i>.

<p>Segments from the chromosome 6 of <i>T. cruzi</i> CL Brener non-Esmeraldo-like and Esmeraldo-like haplotypes, corresponding to TcIII and TcII, respectively, showing 3 to 4 cruzipain gene copies (entire or partial) flanked by orthologous genes marked with different colors according to the legend. Data from the draft assembly of <i>T. cruzi</i> G, M6241 cl6 and <i>T. dionisii</i> allowed to place one, three or two cruzipain gene copies, respectively, within the same syntenic region (figure do not reflect their actual position on chromosomes). Syntenic region from the chromosome 6 of <i>T. b. brucei</i> comprising 11 copies in tandem of brucipain genes was included in the alignment. The shades of vertical gray bars indicate the variable degrees of divergence between sequences according to the legend. The accession codes of all contigs/scaffolds and GenBank accession numbers (in bold) are presented below the corresponding sequences.</p

Network genealogies of predicted amino acid sequences from all domains of genes encoding cruzipain in <i>T. cruzi</i> and homologues in other trypanosome species.

<p>Networks produced using the Neighbour-Net algorithm in SplitsTree v4.11.3, excluding all conserved sites and with Uncorrected p-distance. Networks were produced using entire sequences (A), pre- and pro-domains (B) or restricted to catalytic domains (C) of cruzipain encoding genes from the different trypanosomes are indicated by different symbols and colors according to the legend. Numbers in nodes correspond to support values estimated by performing 100 bootstrap replicates using the same parameter optimized for network inferences.</p

Network and polymorphism analyses on catalytic domain of cruzipain genes from different trypanosome species.

<p>Genes from <i>Schizotrypanum</i> species (<i>T. cruzi, T. c. marinkellei</i> and <i>T. dionisii</i>) were compared with homologues from their closest relative species, <i>T. rangeli,</i> and the distant related <i>T. b. brucei</i>. (A) Network of 33 amino acid predicted sequences constructed using the Neighbour-Net algorithm excluding all conserved sites and with Uncorrected p-distance. The numbers in nodes correspond to bootstrap values from 100 replicates. (B) Polymorphism on cruzipain amino acid sequences from the distinct trypanosome species.</p

<i>Trypanosoma cruzi</i> isolates of all DTUs (TcI-TcVI) and other trypanosome species, and their respective sequences of cruzipain and homologous genes determined in this study or retrieved from data banks.

a<p>TCC, Code number of the isolates/strains cryopreserved in the Trypanosomatid Culture Collection (TCC); Sequences from cruzipain and homologous genes:</p>b<p>whole genes,</p>c<p>catalytic domains;</p>d<p>cruzipain sequences obtained by sequencing of PCR-amplified genes or from genome databases: GenBank, TriTrypDB and drafts genomes from our Tree of Life project or from the Washington University School of Medicine project*;</p>e<p>lineages of <i>T. rangeli</i> defined by Maia da Silva et al. <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0038385#pone.0038385-MaiadaSilva2" target="_blank">[22]</a>, <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0038385#pone.0038385-Freitas1" target="_blank">[61]</a>.</p

Comparative expression analysis of cruzipain and homologues in <i>T. cruzi, T. cruzi marinkellei, T. dionisii</i> and <i>T. rangeli.</i>

<p>(A) Northern blotting analysis of cruzipain transcripts from <i>T. cruzi</i> (Y) and cross-hybridization using the probe consisting of PCR-amplified catalytic domains of <i>T. cruzi</i> Y cruzipain labeled with ³²P T. cruzi Y (TcY-CatL probe); agarose gel of RNA used for this analysis was stained with ethidium bromide (EtBr). (B) CATL proteolytic activities detected in epimastigote lysates of <i>T. cruzi</i> G, <i>T. cruzi marinkellei, T. dionisii</i> and <i>T. rangeli</i>. Activity banding profiles detected in gelatin gels, pH 5.0 and 5 mM DTT were inhibited in gel incubated with 10 µM E-64.</p

Additional file 1: Table S1. of New insights into the evolution of the Trypanosoma cruzi clade provided by a new trypanosome species tightly linked to Neotropical Pteronotus bats and related to an Australian lineage of trypanosomes

Prevalence of Trypanosoma wauwau and geographical origin of Pteronotus spp. examined in this study. (DOC 114 kb

Bats, Trypanosomes, and Triatomines in Ecuador: New Insights into the Diversity, Transmission, and Origins of <i>Trypanosoma cruzi</i> and Chagas Disease

<div><p>The generalist parasite <i>Trypanosoma cruzi</i> has two phylogenetic lineages associated almost exclusively with bats—<i>Trypanosoma cruzi</i> Tcbat and the subspecies <i>T</i>. <i>c</i>. <i>marinkellei</i>. We present new information on the genetic variation, geographic distribution, host associations, and potential vectors of these lineages. We conducted field surveys of bats and triatomines in southern Ecuador, a country endemic for Chagas disease, and screened for trypanosomes by microscopy and PCR. We identified parasites at species and genotype levels through phylogenetic approaches based on 18S ribosomal RNA (18S rRNA) and cytochrome b (cytb) genes and conducted a comparison of nucleotide diversity of the cytb gene. We document for the first time <i>T</i>. <i>cruzi</i> Tcbat and <i>T</i>. <i>c</i>. <i>marinkellei</i> in Ecuador, expanding their distribution in South America to the western side of the Andes. In addition, we found the triatomines <i>Cavernicola pilosa</i> and <i>Triatoma dispar</i> sharing shelters with bats. The comparisons of nucleotide diversity revealed a higher diversity for <i>T</i>. <i>c</i>. <i>marinkellei</i> than any of the <i>T</i>. <i>c</i>. <i>cruzi</i> genotypes associated with Chagas disease. Findings from this study increased both the number of host species and known geographical ranges of both parasites and suggest potential vectors for these two trypanosomes associated with bats in rural areas of southern Ecuador. The higher nucleotide diversity of <i>T</i>. <i>c</i>. <i>marinkellei</i> supports a long evolutionary relationship between <i>T</i>. <i>cruzi</i> and bats, implying that bats are the original hosts of this important parasite.</p></div