American legal discourse on child trafficking: the re/production of inequalities and persistence of child criminalization

The criminalization of children commercially-sexually exploited through prostitution persists despite trafficking laws recognizing this as one of the worst forms of exploitation committed against the most vulnerable social group. This thesis examines the re/production of inequalities in American legal discourse on child trafficking, and why child criminalization persists in this context. Employing a child-centered framework built from multi-conscious feminism and the sociologies of law and childhood, it examines mechanisms of othering and criminalization in key legislative debates, statutes and cases of the United States generally as well as two states exemplifying the retributive and child-protective modes of handling child trafficking. It identifies three themes or issues often presented as binaries that structure child trafficking discourse—adult/child, victim/offender and consent/non-consent—and examines how these are deployed to penalize children in general, and minority and immigrant children in particular. First, processes of marginalization related to race, class, gender and immigration have been vital to the construction of childhood (as normative/deviant) in and through trafficking and prostitution laws, which are reproduced through different types of discourses in both states. Second, both retributive and child-protective modes of response preserve child criminalization by maintaining the tension between prostitution and trafficking, and the female culpability associated with prostitution, including through the denial of the victimization of “repeat offenders.” Finally, despite its prohibition, prostitution is conceptualized in contractual terms, which imputes consent to identities constructed through this discourse and renders commercial-sexual exploitation as merely or primarily involving acts of sale, purchase and consumption

Role of Kif15 and its novel mitotic partner KBP in K-fiber dynamics and chromosome alignment

<div><p>Faithful segregation of the genetic material during the cell cycle is key for the continuation of life. Central to this process is the assembly of a bipolar spindle that aligns the chromosomes and segregates them to the two daughter cells. Spindle bipolarity is strongly dependent on the activity of the homotetrameric kinesin Eg5. However, another kinesin, Kif15, also provides forces needed to separate the spindle poles during prometaphase and to maintain spindle bipolarity at metaphase. Here we identify KBP as a specific interaction partner of Kif15 in mitosis. We show that KBP promotes the localization of Kif15 to the spindle equator close to the chromosomes. Both Kif15 and KBP are required for the alignment of all the chromosomes to the metaphase plate and the assembly of stable kinetochore fibers of the correct length. Taken together our data uncover a novel role for Kif15 in complex with KBP during mitosis.</p></div

The current appearance of speedwell (Veronica filiformis) in the southern part of Bohemia

The thesis summarizes the data about distribution area of speedwell (Veronica filiformis) in South Bohemia and provides their comparison with literary sources. These data are summed up, and both the proven and questionable information are verified in the field. After the revision there were 45 identified and mapped areas out of the 54 originally recorded. The study areas were looked up in herbaria, databases, literature and from professional or amateur botanists. The occurence of Veronica filiformis was confirmed in 23 original and 10 new habitats. These habitats provided us with: coordinates and altitude, estimated size of the area, grafical decsription of population and description of biotope. The dominant species close to the target species were recorded. After the verification a new map field of Veronica filiformis was generated. The individuals of Veronica filiformis were examined in terms of basic morphometric characteristic - the ratio of supporting bracts and peduncle belonging to them

Biophysical characterisation of Aurora-A binding to TACC3act and comparison with TPX2^1-43.

<p>(A) Co-precipitation assay between GST-AurA-DN and TACC3act-H6c and His₆-TPX2^1-43. 5 μM TACC3act-H6c was used in reactions with 1, 2, 5, 10, 20 and 50 μM His₆-TPX2^1-43 (black triangle). GST was used as a binding control. (B) ¹H-¹⁵N HSQC spectra of ¹⁵N-labelled TACC3act in the absence (black) and presence (red) of AurA-DN. TACC3act residues are labelled and chemical shift changes observed on interaction with AurA-DN are marked with arrows. (C) Summary of NMR data mapped onto the primary sequence of TACC3act. Changes in backbone chemical shifts (Δδ) upon binding to Aurora-A are shown on a scale from low (white) to high (red), based on <a href="http://www.plosgenetics.org/article/info:doi/10.1371/journal.pgen.1005345#pgen.1005345.s001" target="_blank">S1C Fig</a>. Secondary structure content is derived from <a href="http://www.plosgenetics.org/article/info:doi/10.1371/journal.pgen.1005345#pgen.1005345.s002" target="_blank">S2 Fig</a>. Chemical shift changes in the presence of TPX2^1-43 are shown on a scale from low (white) to high (blue) using data in <a href="http://www.plosgenetics.org/article/info:doi/10.1371/journal.pgen.1005345#pgen.1005345.s001" target="_blank">S1D Fig</a>. No data is available for P528 (coloured grey) because it lacks a backbone N-H. (D) ¹H-¹⁵N HSQC spectra of ¹⁵N-labelled TACC3act in the presence of AurA-DN (red) and on the addition of TPX2^1-43 (blue). TACC3act residues are labelled and chemical shift changes observed on interaction with TPX2^1-43 are marked with arrows. (E) TACC3act chemical shift changes associated with increasing concentrations of AurA-DN were monitored in the absence (left) or presence (right) of TPX2^1-43 and fit to <a href="http://www.plosgenetics.org/article/info:doi/10.1371/journal.pgen.1005345#pgen.1005345.e001" target="_blank">Eq 1</a>. (F) Binding affinity of TACC3act for AurA-DN in the absence and presence of TPX2^1-43 as determined in Fig 2E. ND, not determined.</p

F543 in TACC3 is required for efficient targeting of TACC3 to the mitotic spindle.

<p>(A) Graphical illustration of domain organization, numbering and properties of key residues in <i>Homo sapiens (Hs)</i> and <i>Gallus gallus (Gg)</i> TACC3 proteins. Framed area below shows properties of mutant TACC3 protein products expressed in F543A, S574A and DEL cells, respectively. (B) Growth curves are shown for WT and mutant cell lines. n = 3 technical replicates, error bars represent standard deviation. (C) Measurement of 5-hydroxymethylcytosine (hmC) by tandem liquid–chromatography mass spectrometry in cells. hmC levels are expressed as parts per million (ppm) of total cytosines or ‘C’. Note that hmC levels inversely correlate with proliferation rate [<a href="http://www.plosgenetics.org/article/info:doi/10.1371/journal.pgen.1005345#pgen.1005345.ref056" target="_blank">56</a>]. n = 3 technical replicates. Statistical significance was assessed using t-test (**** P < 0.0001). (D) Western blot shows TACC3 protein levels in the DT40 cell lines with genotypes as indicated. The p150 subunit of dynactin serves as loading control. (E) TACC3 localisation to the mitotic spindle is impaired in all three mutant DT40 cell lines. In merged images α-tubulin is green, TACC3 is red and DNA is blue. Scale bar = 5 μm. Box plot on right depicts intensity of TACC3 staining on the mitotic spindle. TACC3 signal intensity was quantified in mitotic spindle volumes defined by α-tubulin staining. A minimum of 60 cells was scored per genotype. Statistical significance was assessed using t-test (**** P < 0.0001).</p

Spatially distinct residues in TACC3 dictate mitotic duration and fidelity.

<p>(A) Spindle length measured in 3D using Volocity. Number of cells analysed is indicated in graph (n). Student t-test (**** P < 0.0001). (B) Mitotic spindle morphologies (as described in main text) observed during time-lapse microscopy of GFP-tubulin-expressing TACC3 mutant cell lines. Representative still frames are shown on left. Scale bar = 5 μm. (C) Durations of NEBD to anaphase onset obtained from time-lapse microscopy performed on GFP-tubulin-expressing TACC3 mutant cell lines. F543A_1 and F543A_2 cells represent independently derived clones. Note that we observed no correlation between GFP levels and mitotic timing, or between mitotic timing and time of NEBD with respect to duration of filming in these experiments. Box plot shows 10–90 percentiles for each genotype. Number of cells analysed is indicated in graph (n). Mann Whitney nonparametric t-test (**** P < 0.0001, *** P < 0.001 and n.s. stands for ‘no significance’). (D) Durations of NEBD to anaphase onset obtained from time-lapse microscopy performed on EB3-GFP-expressing cells. Box plot shows 10–90 percentiles. Number of cells analysed is indicated in the graph (n). Mann Whitney nonparametric t-test. (**** P < 0.0001). (E) Analysis of chromosome segregation. Top table shows percentage of lagging chromatids seen during anaphase in fixed cells. Bottom table shows results from metaphase chromosome (chr) spreads. Number of autosomes 1, 2, 3, 4 and the sex chromosome, Z was analysed in cells. Frequencies of loss or gain of a single copy of individual chromosomes are indicated. Note that WT DT40 cells are trisomic for chromosome 2, whilst all the TACC3 mutants are diploid. Therefore, cells with two or three copies of chromosome 2’s are marked with ‘*’ or ‘#’, respectively. (F) Plot depicts time between NEBD and bipolar spindle appearance in time-lapse microscopy from Fig 7B. Cells with multipolar spindles and cells in which one of the two spindle poles was outside of the Z stack range for these time points were excluded from analysis. Number of cells analysed is indicated in graph (n). Mann Whitney nonparametric t-test (* P = 0.04, **** P < 0.0001).</p