Commit e74fe8cf authored by mk11g11's avatar mk11g11
Browse files

corrections

parent 9e297c11
fig/results5/png/Hua7_expression.png

241 KB | W: | H:

fig/results5/png/Hua7_expression.png

170 KB | W: | H:

fig/results5/png/Hua7_expression.png
fig/results5/png/Hua7_expression.png
fig/results5/png/Hua7_expression.png
fig/results5/png/Hua7_expression.png
  • 2-up
  • Swipe
  • Onion skin
......@@ -2,7 +2,7 @@
# YAML for thesis template
site: bookdown::bookdown_site
documentclass: book
bibliography: [bib/packages.bib, bib/papers.bib, bib/august.bib] # Changes to reference bib files as needed
bibliography: [bib/packages.bib, bib/papers.bib, bib/march.bib] # Changes to reference bib files as needed
biblio-style: apalike
toc: False # Table of contents supplied from tex files
lof: False # List of figures supplied from tex files
......
\addcontentsline{toc}{chapter}{List of Abbreviations}
\chapter*{List of Abbreviations}
nAChR : nicotinic acetylcholine receptor \\
ECD : extracellular domain \\
BWM: Body Wall Muscle \\
5-HT - 5-hydroxytryptamine \\
ACE - Acetylcholinesterase \\
AChBP - Acetylcholine Binding Protein \\
bgtx - bungarotoxin \\
RIC-3 - Resistant to Inhibitors of Cholinesterase-3 \\
GPCR - G protein coupled receptor \\
Bovine Serum Albumin (BSA) \\
e. coli
polyvinylidene difluoride (PVDF)
C. elegans
GLIC - \textit{Gloebacter} ligand gated ion channels \\
Green Fluorescent Protein (GFP) \\
BWM - Body Wall Muscle \\
Ch - choline \\
ChAt - Choline acetyltransferase \\
ACE - Acetylcholinesterase \\
VAChT - Vesicular acetylcholine transferase \\
\ No newline at end of file
dimethyl sulfoxide (DMSO) \\
ECD - extracellular domain \\
E/R ratio
ELIC - \textit{Erwinia} ligand gated ion channels
GABA - Gamma aminobutyric acid \\
GPCR - G protein coupled receptor \\
nAChR : nicotinic acetylcholine receptor \\
NGM - nematode growth medium \\
RIC-3 - Resistant to Inhibitors of Cholinesterase-3 \\
TM - transmembrane
VAChT - Vesicular acetylcholine transferase \\
MBP - Maltose Binding Protein
\ No newline at end of file
\addcontentsline{toc}{chapter}{Abstract}
\chapter*{Abstract}
An ability of insecticides to selectively target pests without affecting non-target species is a key determinant of success of componds used in agriculture. Neonicotinoids are a major class of insecticide which encompases seven chemical representing three distinct chemical classes, namely cyanoamidines, nitroguanidines and nitromethylenes. Neonicotinoids effectively control a wide range of insect pests and have low toxicity against mammals, however they can also negatively impact on non-target species of bees, threatening food safety. Neonicotinoids act by targeting insect nAChR, which are a major excitatory receptors in the insect central nervous system. Difficulties in heterologous expression of these proteins hinders their pharmacological characterisation and identification of molecular determinants of neonicotinoid-toxicity. This thesis describes the efforts into developing \textit{C. elegans}* as a platfom in which the mode of action and selective toxicity of neonicotinoid-insecticides can be studied.
An ability of insecticides to selectively target pests without affecting non-target species is key determinant of success of compounds used in agriculture. Neonicotinoids which encompass seven different types of chemical representing three distinct chemical classes, namely the cyanoamidines, nitroguanidines and nitromethylenes, are a major class of insecticides. They effectively control a wide range of insect pests and have low toxicity against mammals, however they can also negatively impact on non-target species of bees, threatening food safety. Neonicotinoids act by targeting insect nicotinic acetylcholine receortors (nAChRs), which are major excitatory receptors in the insect central nervous system. Difficulties in heterologous expression of these proteins hinders their pharmacological characterisation and identification of the molecular determinants of neonicotinoid-toxicity. This thesis describes efforts into developing \textit{Caenorhabditis elegans (C. elegans)} as a platform in which the mode of action and selective toxicity of neonicotinoid-insecticides can be studied.
We determined the effects of neonicotinoids on \textit{C. elegans} behaviours governed by the cholinergic neurotransmission. Cyanoamidines clothianidin, nitroguanidine thiacloprid and nitromethylene nitenpyram showed low efficacy on locomotion, pharyngeal pumping, egg-laying and egg-hatching of young adult wild-type \textit{C. elegans}. Exposure of mutant worm with enhanced cuticle permeability showed increased susceptibility of worms to all three neonicotinoids tested, suggesting an adult cuticle limits drug access. The role of the cuticle in neonicotinoids susceptibility was nvestigated in *C. elegans* cut-head preparation, in which the cuticle is removed and the effects of compounds on pharyngeal pumping are scored. Out of the three neonicotinoids applied, clothianidin showed the greatest efficacy; it stimulated pharyngeal pumping at $\geq$ 75 $\mu$M. The concentrations effective against the function of the pharynx are an order or magnitude lower than the residual, average concenntration in the soil, suggesting *C. elegans* is not impacted in the field, and at least 8-fold lower than lethal doses in insect-pests. The difference in neonicotinoid-susceptiility between adult \textit{C. elegans} and insects precludes the use of *C. elegans* pharynx as a platform for the mode of action studies, but highlights its potential as a suitable background for the heterologous expression of insect nAChRs.
We determined the effects of neonicotinoids on \textit{C. elegans} behaviours governed by the cholinergic neurotransmission. The cyanoamidine represented by clothianidin, the nitroguanidine represented by thiacloprid and the nitromethylene represented by nitenpyram showed low efficacy on locomotion, pharyngeal pumping, egg-laying and egg-hatching of young adult wild-type \textit{C. elegans}. Exposure of mutant worm with enhanced cuticular permeability showed increased susceptibility of worms to all three neonicotinoids tested, suggesting an adult cuticle limits drug access. The role of the cuticle in neonicotinoids susceptibility was investigated in \textit{C. elegans} cut-head preparation, in which the cuticle is removed and the effects of compounds on pharyngeal pumping are scored. Out of the three neonicotinoids applied, clothianidin showed the greatest efficacy. It stimulated pharyngeal pumping at $\geq$ 75 $\mu$M (18.75 ppm). Generally, the concentrations effective against the function of the pharynx are an order or magnitude lower than the residual, average concentration of neonicotinoids in the soil, suggesting \textit{C. elegans} is not impacted in the field, and at least several fold lower than lethal doses in insect-pests. The difference in neonicotinoid-susceptibility between adult \textit{C. elegans} and insects precludes the use of \textit{C. elegans} pharynx as a platform for the mode of action studies, but highlights its potential as a suitable background for the heterologous expression of insect nAChRs.
Further experiments show that \textit{C. elegans eat-2} nAChR mutant is a suitable genetic background, in which the expression of heterologous nAChRs can be scored in behavioural and pharmacological assays. Expression of nAChR EAT-2 in the pharyngeal muscle rescued the blunted feeding phenotype and 5-HT insensitivity of the \textit{eat-2} mutant worm. Expression of exogenous receptor, viz. human $\alpha$7 in the pharynx of \textit{eat-2} mutant led to a cell-surface expression, as shown by staining with labelled $\alpha$-bungarotoxin. However the feeding and pharmacological phenotypes of the mutant were not rescued. \textit{C. elegans} strains in which human $\alpha$7 is expressed in the wild-type genetic background was also generated to determine whether the pharmacology of the human receptor can be inprinted on the \textit{C. elegans} pharynx. No difference in the phayngeal response to nAChR agonists cytisine, nicotine or acetylcholine were noted. The lack of apparent functionality of $\alpha$7 receptor could be due to the incorrect cellular localisation of this protein. $\alpha$-bgtx staining showed that the expression of $\alpha$7 receptor is concentrated to the specific cells of the pharyngeal muscle, however this localisation does not overlap with the localisation of native EAT-2 receptors. A transgenic strains in which exogenous proteins are expressed using EAT-2 native promoter should be made.
\ No newline at end of file
Further experiments showed that \textit{C. elegans eat-2} nAChR mutant is a suitable genetic background, in which the expression of heterologous nAChRs can be scored. Expression of \textit{C. elegans} nAChR EAT-2 in the pharyngeal muscle rescued the blunted feeding phenotype and 5-HT insensitivity of the \textit{eat-2} mutant. Expression of the exogenous receptor, human $\alpha$7 in the pharynx of \textit{eat-2} mutant led to a cell-surface expression, as shown by staining with labelled $\alpha$-bungarotoxin. However the feeding and pharmacological phenotypes of the mutant were not rescued. \textit{C. elegans} strain in which human $\alpha$7 is expressed in the wild-type genetic background was also generated to determine whether the pharmacology of the human receptor can be imposed on the \textit{C. elegans} pharynx. No difference in the phayngeal response to nAChR agonists cytisine, nicotine or acetylcholine were noted. The lack of apparent functionality of $\alpha$7 receptor could be due to the incorrect cellular localisation of this protein. $\alpha$-bgtx staining showed that $\alpha$7 receptor is expressed in the specific cells of the pharyngeal muscle, however this localisation does not overlap with the localisation of native EAT-2 receptors. A transgenic strain in which exogenous proteins are expressed using EAT-2 native promoter should be made.
\ No newline at end of file
......@@ -3,12 +3,6 @@
Part of this thesis has been published in the following papers:
Kudelska, M. M., L. Holden-Dye, V. O’Connor, and D. A. Doyle. 2017. “Concentration-Dependent Effects
of Acute and Chronic Neonicotinoid Exposure on the Behaviour and Development of the Nematode
Caenorhabditis Elegans.” Journal Article. Pest Manag Sci 73 (7): 1345–51. https://doi.org/10.1002/ps.
4564.
Kudelska, M. M., L. Holden-Dye, V. O'Connor, and D. A. Doyle. 2017. Concentration-Dependent Effects of Acute and Chronic Neonicotinoid Exposure on the Behaviour and Development of the Nematode \textit{Caenorhabditis elegans}. Journal Article. Pest Manag Sci 73 (7): 1345–51. https://doi.org/10.1002/ps.4564.
Kudelska, M. M., A. Lewis, C. T. Ng, D. A. Doyle, L. Holden-Dye, V. M. O’Connor, and R. J. Walker.
2018. “Investigation of Feeding Behaviour in c. Elegans Reveals Distinct Pharmacological
and Antibacterial Effects of Nicotine.” Invertebrate Neuroscience : IN 18 (4): 14. https:
//doi.org/10.1007/s10158-018-0219-1.
\ No newline at end of file
Kudelska, M. M., A. Lewis, C. T. Ng, D. A. Doyle, L. Holden-Dye, V. M. O'Connor, and R. J. Walker. 2018. Investigation of Feeding Behaviour in \textit{c. elegans} Reveals Distinct Pharmacological and Antibacterial Effects of Nicotine. Invertebrate Neuroscience : IN 18 (4): 14. https://doi.org/10.1007/s10158-018-0219-1.
\ No newline at end of file
......@@ -26,8 +26,8 @@
{\rm\hfill\sl\leftmark}}}
%\def\@oddfoot{\thepage}
%\def\@evenfoot{\thepage}
\def\@evenhead{\protect\underline{\protect\makebox[\textwidth][l]
{\rm\hfill\sl\leftmark}}}
% \def\@evenhead{\protect\underline{\protect\makebox[\textwidth][l]
% {\rm\hfill\sl\leftmark}}}
% end of modifications
\def\chaptermark##1{\markboth {\ifnum \c@secnumdepth >\m@ne
\chapapp2\ \thechapter. \ \fi ##1}{}}%
......
This is XeTeX, Version 3.14159265-2.6-0.999991 (TeX Live 2019/W32TeX) (preloaded format=xelatex 2019.8.18) 6 OCT 2019 15:49
entering extended mode
restricted \write18 enabled.
%&-line parsing enabled.
**thesis-dissertaion.tex
(./thesis-dissertaion.tex
LaTeX2e <2018-12-01>
(c:/Users/monik/AppData/Roaming/TinyTeX/texmf-dist/tex/latex/base/book.cls
Document Class: book 2018/09/03 v1.4i Standard LaTeX document class
(c:/Users/monik/AppData/Roaming/TinyTeX/texmf-dist/tex/latex/base/bk10.clo
File: bk10.clo 2018/09/03 v1.4i Standard LaTeX file (size option)
)
\c@part=\count80
\c@chapter=\count81
\c@section=\count82
\c@subsection=\count83
\c@subsubsection=\count84
\c@paragraph=\count85
\c@subparagraph=\count86
\c@figure=\count87
\c@table=\count88
\abovecaptionskip=\skip41
\belowcaptionskip=\skip42
\bibindent=\dimen102
) (c:/Users/monik/AppData/Roaming/TinyTeX/texmf-dist/tex/latex/lm/lmodern.sty
Package: lmodern 2009/10/30 v1.6 Latin Modern Fonts
LaTeX Font Info: Overwriting symbol font `operators' in version `normal'
(Font) OT1/cmr/m/n --> OT1/lmr/m/n on input line 22.
LaTeX Font Info: Overwriting symbol font `letters' in version `normal'
(Font) OML/cmm/m/it --> OML/lmm/m/it on input line 23.
LaTeX Font Info: Overwriting symbol font `symbols' in version `normal'
(Font) OMS/cmsy/m/n --> OMS/lmsy/m/n on input line 24.
LaTeX Font Info: Overwriting symbol font `largesymbols' in version `normal'
(Font) OMX/cmex/m/n --> OMX/lmex/m/n on input line 25.
LaTeX Font Info: Overwriting symbol font `operators' in version `bold'
(Font) OT1/cmr/bx/n --> OT1/lmr/bx/n on input line 26.
LaTeX Font Info: Overwriting symbol font `letters' in version `bold'
(Font) OML/cmm/b/it --> OML/lmm/b/it on input line 27.
LaTeX Font Info: Overwriting symbol font `symbols' in version `bold'
(Font) OMS/cmsy/b/n --> OMS/lmsy/b/n on input line 28.
LaTeX Font Info: Overwriting symbol font `largesymbols' in version `bold'
(Font) OMX/cmex/m/n --> OMX/lmex/m/n on input line 29.
LaTeX Font Info: Overwriting math alphabet `\mathbf' in version `normal'
(Font) OT1/cmr/bx/n --> OT1/lmr/bx/n on input line 31.
LaTeX Font Info: Overwriting math alphabet `\mathsf' in version `normal'
(Font) OT1/cmss/m/n --> OT1/lmss/m/n on input line 32.
LaTeX Font Info: Overwriting math alphabet `\mathit' in version `normal'
(Font) OT1/cmr/m/it --> OT1/lmr/m/it on input line 33.
LaTeX Font Info: Overwriting math alphabet `\mathtt' in version `normal'
(Font) OT1/cmtt/m/n --> OT1/lmtt/m/n on input line 34.
LaTeX Font Info: Overwriting math alphabet `\mathbf' in version `bold'
(Font) OT1/cmr/bx/n --> OT1/lmr/bx/n on input line 35.
LaTeX Font Info: Overwriting math alphabet `\mathsf' in version `bold'
(Font) OT1/cmss/bx/n --> OT1/lmss/bx/n on input line 36.
LaTeX Font Info: Overwriting math alphabet `\mathit' in version `bold'
(Font) OT1/cmr/bx/it --> OT1/lmr/bx/it on input line 37.
LaTeX Font Info: Overwriting math alphabet `\mathtt' in version `bold'
(Font) OT1/cmtt/m/n --> OT1/lmtt/m/n on input line 38.
)
(c:/Users/monik/AppData/Roaming/TinyTeX/texmf-dist/tex/latex/amsfonts/amssymb.s
ty
Package: amssymb 2013/01/14 v3.01 AMS font symbols
(c:/Users/monik/AppData/Roaming/TinyTeX/texmf-dist/tex/latex/amsfonts/amsfonts.
sty
Package: amsfonts 2013/01/14 v3.01 Basic AMSFonts support
\@emptytoks=\toks14
\symAMSa=\mathgroup4
\symAMSb=\mathgroup5
LaTeX Font Info: Overwriting math alphabet `\mathfrak' in version `bold'
(Font) U/euf/m/n --> U/euf/b/n on input line 106.
))
(c:/Users/monik/AppData/Roaming/TinyTeX/texmf-dist/tex/latex/amsmath/amsmath.st
y
Package: amsmath 2018/12/01 v2.17b AMS math features
\@mathmargin=\skip43
For additional information on amsmath, use the `?' option.
(c:/Users/monik/AppData/Roaming/TinyTeX/texmf-dist/tex/latex/amsmath/amstext.st
y
Package: amstext 2000/06/29 v2.01 AMS text
(c:/Users/monik/AppData/Roaming/TinyTeX/texmf-dist/tex/latex/amsmath/amsgen.sty
File: amsgen.sty 1999/11/30 v2.0 generic functions
\@emptytoks=\toks15
\ex@=\dimen103
))
(c:/Users/monik/AppData/Roaming/TinyTeX/texmf-dist/tex/latex/amsmath/amsbsy.sty
Package: amsbsy 1999/11/29 v1.2d Bold Symbols
\pmbraise@=\dimen104
)
(c:/Users/monik/AppData/Roaming/TinyTeX/texmf-dist/tex/latex/amsmath/amsopn.sty
Package: amsopn 2016/03/08 v2.02 operator names
)
\inf@bad=\count89
LaTeX Info: Redefining \frac on input line 223.
\uproot@=\count90
\leftroot@=\count91
LaTeX Info: Redefining \overline on input line 385.
\classnum@=\count92
\DOTSCASE@=\count93
LaTeX Info: Redefining \ldots on input line 482.
LaTeX Info: Redefining \dots on input line 485.
LaTeX Info: Redefining \cdots on input line 606.
\Mathstrutbox@=\box27
\strutbox@=\box28
\big@size=\dimen105
LaTeX Font Info: Redeclaring font encoding OML on input line 729.
LaTeX Font Info: Redeclaring font encoding OMS on input line 730.
\macc@depth=\count94
\c@MaxMatrixCols=\count95
\dotsspace@=\muskip10
\c@parentequation=\count96
\dspbrk@lvl=\count97
\tag@help=\toks16
\row@=\count98
\column@=\count99
\maxfields@=\count100
\andhelp@=\toks17
\eqnshift@=\dimen106
\alignsep@=\dimen107
\tagshift@=\dimen108
\tagwidth@=\dimen109
\totwidth@=\dimen110
\lineht@=\dimen111
\@envbody=\toks18
\multlinegap=\skip44
\multlinetaggap=\skip45
\mathdisplay@stack=\toks19
LaTeX Info: Redefining \[ on input line 2844.
LaTeX Info: Redefining \] on input line 2845.
)
(c:/Users/monik/AppData/Roaming/TinyTeX/texmf-dist/tex/generic/ifxetex/ifxetex.
sty
Package: ifxetex 2010/09/12 v0.6 Provides ifxetex conditional
)
(c:/Users/monik/AppData/Roaming/TinyTeX/texmf-dist/tex/generic/oberdiek/ifluate
x.sty
Package: ifluatex 2016/05/16 v1.4 Provides the ifluatex switch (HO)
Package ifluatex Info: LuaTeX not detected.
) (c:/Users/monik/AppData/Roaming/TinyTeX/texmf-dist/tex/latex/base/fixltx2e.st
y
Package: fixltx2e 2016/12/29 v2.1a fixes to LaTeX (obsolete)
Applying: [2015/01/01] Old fixltx2e package on input line 46.
Package fixltx2e Warning: fixltx2e is not required with releases after 2015
(fixltx2e) All fixes are now in the LaTeX kernel.
(fixltx2e) See the latexrelease package for details.
Already applied: [0000/00/00] Old fixltx2e package on input line 53.
)
(c:/Users/monik/AppData/Roaming/TinyTeX/texmf-dist/tex/xelatex/mathspec/mathspe
c.sty
Package: mathspec 2016/12/22 v0.2b LaTeX Package (Mathematics font selection fo
r XeLaTeX)
(c:/Users/monik/AppData/Roaming/TinyTeX/texmf-dist/tex/latex/etoolbox/etoolbox.
sty
Package: etoolbox 2018/08/19 v2.5f e-TeX tools for LaTeX (JAW)
\etb@tempcnta=\count101
)
(c:/Users/monik/AppData/Roaming/TinyTeX/texmf-dist/tex/latex/fontspec/fontspec.
sty
(c:/Users/monik/AppData/Roaming/TinyTeX/texmf-dist/tex/latex/l3packages/xparse/
xparse.sty
(c:/Users/monik/AppData/Roaming/TinyTeX/texmf-dist/tex/latex/l3kernel/expl3.sty
Package: expl3 2019-08-14 L3 programming layer (loader)
(c:/Users/monik/AppData/Roaming/TinyTeX/texmf-dist/tex/latex/l3kernel/expl3-cod
e.tex
Package: expl3 2019-08-14 L3 programming layer (code)
\c_max_int=\count102
\l_tmpa_int=\count103
\l_tmpb_int=\count104
\g_tmpa_int=\count105
\g_tmpb_int=\count106
\g__kernel_prg_map_int=\count107
\c__ior_term_noprompt_ior=\count108
\c_log_iow=\count109
\l_iow_line_count_int=\count110
\l__iow_line_target_int=\count111
\l__iow_one_indent_int=\count112
\l__iow_indent_int=\count113
\c_zero_dim=\dimen112
\c_max_dim=\dimen113
\l_tmpa_dim=\dimen114
\l_tmpb_dim=\dimen115
\g_tmpa_dim=\dimen116
\g_tmpb_dim=\dimen117
\c_zero_skip=\skip46
\c_max_skip=\skip47
\l_tmpa_skip=\skip48
\l_tmpb_skip=\skip49
\g_tmpa_skip=\skip50
\g_tmpb_skip=\skip51
\c_zero_muskip=\muskip11
\c_max_muskip=\muskip12
\l_tmpa_muskip=\muskip13
\l_tmpb_muskip=\muskip14
\g_tmpa_muskip=\muskip15
\g_tmpb_muskip=\muskip16
\l_keys_choice_int=\count114
\l__intarray_loop_int=\count115
\c__intarray_sp_dim=\dimen118
\g__intarray_font_int=\count116
\c__fp_leading_shift_int=\count117
\c__fp_middle_shift_int=\count118
\c__fp_trailing_shift_int=\count119
\c__fp_big_leading_shift_int=\count120
\c__fp_big_middle_shift_int=\count121
\c__fp_big_trailing_shift_int=\count122
\c__fp_Bigg_leading_shift_int=\count123
\c__fp_Bigg_middle_shift_int=\count124
\c__fp_Bigg_trailing_shift_int=\count125
\g__fp_array_int=\count126
\l__fp_array_loop_int=\count127
\l__sort_length_int=\count128
\l__sort_min_int=\count129
\l__sort_top_int=\count130
\l__sort_max_int=\count131
\l__sort_true_max_int=\count132
\l__sort_block_int=\count133
\l__sort_begin_int=\count134
\l__sort_end_int=\count135
\l__sort_A_int=\count136
\l__sort_B_int=\count137
\l__sort_C_int=\count138
\l__str_internal_int=\count139
\l__tl_analysis_normal_int=\count140
\l__tl_analysis_index_int=\count141
\l__tl_analysis_nesting_int=\count142
\l__tl_analysis_type_int=\count143
\l__regex_internal_a_int=\count144
\l__regex_internal_b_int=\count145
\l__regex_internal_c_int=\count146
\l__regex_balance_int=\count147
\l__regex_group_level_int=\count148
\l__regex_mode_int=\count149
\c__regex_cs_in_class_mode_int=\count150
\c__regex_cs_mode_int=\count151
\l__regex_catcodes_int=\count152
\l__regex_default_catcodes_int=\count153
\c__regex_catcode_L_int=\count154
\c__regex_catcode_O_int=\count155
\c__regex_catcode_A_int=\count156
\c__regex_all_catcodes_int=\count157
\l__regex_show_lines_int=\count158
\l__regex_min_state_int=\count159
\l__regex_max_state_int=\count160
\l__regex_left_state_int=\count161
\l__regex_right_state_int=\count162
\l__regex_capturing_group_int=\count163
\l__regex_min_pos_int=\count164
\l__regex_max_pos_int=\count165
\l__regex_curr_pos_int=\count166
\l__regex_start_pos_int=\count167
\l__regex_success_pos_int=\count168
\l__regex_curr_char_int=\count169
\l__regex_curr_catcode_int=\count170
\l__regex_last_char_int=\count171
\l__regex_case_changed_char_int=\count172
\l__regex_curr_state_int=\count173
\l__regex_step_int=\count174
\l__regex_min_active_int=\count175
\l__regex_max_active_int=\count176
\l__regex_replacement_csnames_int=\count177
\l__regex_match_count_int=\count178
\l__regex_min_submatch_int=\count179
\l__regex_submatch_int=\count180
\l__regex_zeroth_submatch_int=\count181
\g__regex_trace_regex_int=\count182
\c_empty_box=\box29
\l_tmpa_box=\box30
\l_tmpb_box=\box31
\g_tmpa_box=\box32
\g_tmpb_box=\box33
\l__box_top_dim=\dimen119
\l__box_bottom_dim=\dimen120
\l__box_left_dim=\dimen121
\l__box_right_dim=\dimen122
\l__box_top_new_dim=\dimen123
\l__box_bottom_new_dim=\dimen124
\l__box_left_new_dim=\dimen125
\l__box_right_new_dim=\dimen126
\l__box_internal_box=\box34
\l__coffin_internal_box=\box35
\l__coffin_internal_dim=\dimen127
\l__coffin_offset_x_dim=\dimen128
\l__coffin_offset_y_dim=\dimen129
\l__coffin_x_dim=\dimen130
\l__coffin_y_dim=\dimen131
\l__coffin_x_prime_dim=\dimen132
\l__coffin_y_prime_dim=\dimen133
\c_empty_coffin=\box36
\l__coffin_aligned_coffin=\box37
\l__coffin_aligned_internal_coffin=\box38
\l_tmpa_coffin=\box39
\l_tmpb_coffin=\box40
\g_tmpa_coffin=\box41
\g_tmpb_coffin=\box42
\l__coffin_bounding_shift_dim=\dimen134
\l__coffin_left_corner_dim=\dimen135
\l__coffin_right_corner_dim=\dimen136
\l__coffin_bottom_corner_dim=\dimen137
\l__coffin_top_corner_dim=\dimen138
\l__coffin_scaled_total_height_dim=\dimen139
\l__coffin_scaled_width_dim=\dimen140
\c__coffin_empty_coffin=\box43
\l__coffin_display_coffin=\box44
\l__coffin_display_coord_coffin=\box45
\l__coffin_display_pole_coffin=\box46
\l__coffin_display_offset_dim=\dimen141
\l__coffin_display_x_dim=\dimen142
\l__coffin_display_y_dim=\dimen143
\g__file_internal_ior=\read1
\l__seq_internal_a_int=\count183
\l__seq_internal_b_int=\count184
)
(c:/Users/monik/AppData/Roaming/TinyTeX/texmf-dist/tex/latex/l3backend/l3backen
d-xdvipdfmx.def
File: l3backend-xdvipdfmx.def 2019-04-06 v L3 Experimental backend support: xdv
ipdfmx
\g__graphics_track_int=\count185
\l__pdf_internal_box=\box47
\g__pdf_backend_object_int=\count186
\g__pdf_backend_annotation_int=\count187
))
Package: xparse 2019-05-28 L3 Experimental document command parser
\l__xparse_current_arg_int=\count188
\g__xparse_grabber_int=\count189
\l__xparse_m_args_int=\count190
\l__xparse_v_nesting_int=\count191
)
Package: fontspec 2019/03/15 v2.7c Font selection for XeLaTeX and LuaLaTeX
(c:/Users/monik/AppData/Roaming/TinyTeX/texmf-dist/tex/latex/fontspec/fontspec-
xetex.sty
Package: fontspec-xetex 2019/03/15 v2.7c Font selection for XeLaTeX and LuaLaTe
X
\l__fontspec_script_int=\count192
\l__fontspec_language_int=\count193
\l__fontspec_strnum_int=\count194
\l__fontspec_tmp_int=\count195
\l__fontspec_tmpa_int=\count196
\l__fontspec_tmpb_int=\count197
\l__fontspec_tmpc_int=\count198
\l__fontspec_em_int=\count199
\l__fontspec_emdef_int=\count266
\l__fontspec_strong_int=\count267
\l__fontspec_strongdef_int=\count268
\l__fontspec_tmpa_dim=\dimen144
\l__fontspec_tmpb_dim=\dimen145
\l__fontspec_tmpc_dim=\dimen146
(c:/Users/monik/AppData/Roaming/TinyTeX/texmf-dist/tex/latex/base/fontenc.sty
Package: fontenc 2018/08/11 v2.0j Standard LaTeX package
(c:/Users/monik/AppData/Roaming/TinyTeX/texmf-dist/tex/latex/base/tuenc.def
File: tuenc.def 2018/08/11 v2.0j Standard LaTeX file
LaTeX Font Info: Redeclaring font encoding TU on input line 82.
))
(c:/Users/monik/AppData/Roaming/TinyTeX/texmf-dist/tex/latex/fontspec/fontspec.
cfg)
LaTeX Info: Redefining \itshape on input line 4051.
LaTeX Info: Redefining \slshape on input line 4056.
LaTeX Info: Redefining \scshape on input line 4061.
LaTeX Info: Redefining \upshape on input line 4066.
LaTeX Info: Redefining \em on input line 4096.
LaTeX Info: Redefining \emph on input line 4121.
))
(c:/Users/monik/AppData/Roaming/TinyTeX/texmf-dist/tex/latex/xkeyval/xkeyval.st
y
Package: xkeyval 2014/12/03 v2.7a package option processing (HA)
(c:/Users/monik/AppData/Roaming/TinyTeX/texmf-dist/tex/generic/xkeyval/xkeyval.
tex
(c:/Users/monik/AppData/Roaming/TinyTeX/texmf-dist/tex/generic/xkeyval/xkvutils
.tex
\XKV@toks=\toks20
\XKV@tempa@toks=\toks21
(c:/Users/monik/AppData/Roaming/TinyTeX/texmf-dist/tex/generic/xkeyval/keyval.t
ex))
\XKV@depth=\count269
File: xkeyval.tex 2014/12/03 v2.7a key=value parser (HA)
))
\c@eu@=\count270
\c@eu@i=\count271
\c@mkern=\count272
)
(c:/Users/monik/AppData/Roaming/TinyTeX/texmf-dist/tex/latex/upquote/upquote.st
y
Package: upquote 2012/04/19 v1.3 upright-quote and grave-accent glyphs in verba
tim
(c:/Users/monik/AppData/Roaming/TinyTeX/texmf-dist/tex/latex/base/textcomp.sty
Package: textcomp 2018/08/11 v2.0j Standard LaTeX package
Package textcomp Info: Sub-encoding information:
(textcomp) 5 = only ISO-Adobe without \textcurrency
(textcomp) 4 = 5 + \texteuro
(textcomp) 3 = 4 + \textohm
(textcomp) 2 = 3 + \textestimated + \textcurrency
(textcomp) 1 = TS1 - \textcircled - \t
(textcomp) 0 = TS1 (full)
(textcomp) Font families with sub-encoding setting implement
(textcomp) only a restricted character set as indicated.
(textcomp) Family '?' is the default used for unknown fonts.
(textcomp) See the documentation for details.
Package textcomp Info: Setting ? sub-encoding to TS1/1 on input line 79.
(c:/Users/monik/AppData/Roaming/TinyTeX/texmf-dist/tex/latex/base/ts1enc.def
File: ts1enc.def 2001/06/05 v3.0e (jk/car/fm) Standard LaTeX file
)
LaTeX Info: Redefining \oldstylenums on input line 334.
Package textcomp Info: Setting cmr sub-encoding to TS1/0 on input line 349.
Package textcomp Info: Setting cmss sub-encoding to TS1/0 on input line 350.
Package textcomp Info: Setting cmtt sub-encoding to TS1/0 on input line 351.
Package textcomp Info: Setting cmvtt sub-encoding to TS1/0 on input line 352.
Package textcomp Info: Setting cmbr sub-encoding to TS1/0 on input line 353.
Package textcomp Info: Setting cmtl sub-encoding to TS1/0 on input line 354.
Package textcomp Info: Setting ccr sub-encoding to TS1/0 on input line 355.
Package textcomp Info: Setting ptm sub-encoding to TS1/4 on input line 356.
Package textcomp Info: Setting pcr sub-encoding to TS1/4 on input line 357.
Package textcomp Info: Setting phv sub-encoding to TS1/4 on input line 358.
Package textcomp Info: Setting ppl sub-encoding to TS1/3 on input line 359.
Package textcomp Info: Setting pag sub-encoding to TS1/4 on input line 360.
Package textcomp Info: Setting pbk sub-encoding to TS1/4 on input line 361.
Package textcomp Info: Setting pnc sub-encoding to TS1/4 on input line 362.
Package textcomp Info: Setting pzc sub-encoding to TS1/4 on input line 363.
Package textcomp Info: Setting bch sub-encoding to TS1/4 on input line 364.
Package textcomp Info: Setting put sub-encoding to TS1/5 on input line 365.
Package textcomp Info: Setting uag sub-encoding to TS1/5 on input line 366.
Package textcomp Info: Setting ugq sub-encoding to TS1/5 on input line 367.
Package textcomp Info: Setting ul8 sub-encoding to TS1/4 on input line 368.
Package textcomp Info: Setting ul9 sub-encoding to TS1/4 on input line 369.
Package textcomp Info: Setting augie sub-encoding to TS1/5 on input line 370.
Package textcomp Info: Setting dayrom sub-encoding to TS1/3 on input line 371.
Package textcomp Info: Setting dayroms sub-encoding to TS1/3 on input line 372.
Package textcomp Info: Setting pxr sub-encoding to TS1/0 on input line 373.
Package textcomp Info: Setting pxss sub-encoding to TS1/0 on input line 374.
Package textcomp Info: Setting pxtt sub-encoding to TS1/0 on input line 375.
Package textcomp Info: Setting txr sub-encoding to TS1/0 on input line 376.
Package textcomp Info: Setting txss sub-encoding to TS1/0 on input line 377.
Package textcomp Info: Setting txtt sub-encoding to TS1/0 on input line 378.
Package textcomp Info: Setting lmr sub-encoding to TS1/0 on input line 379.
Package textcomp Info: Setting lmdh sub-encoding to TS1/0 on input line 380.
Package textcomp Info: Setting lmss sub-encoding to TS1/0 on input line 381.
Package textcomp Info: Setting lmssq sub-encoding to TS1/0 on input line 382.
Package textcomp Info: Setting lmvtt sub-encoding to TS1/0 on input line 383.
Package textcomp Info: Setting lmtt sub-encoding to TS1/0 on input line 384.
Package textcomp Info: Setting qhv sub-encoding to TS1/0 on input line 385.
Package textcomp Info: Setting qag sub-encoding to TS1/0 on input line 386.
Package textcomp Info: Setting qbk sub-encoding to TS1/0 on input line 387.
Package textcomp Info: Setting qcr sub-encoding to TS1/0 on input line 388.
Package textcomp Info: Setting qcs sub-encoding to TS1/0 on input line 389.
Package textcomp Info: Setting qpl sub-encoding to TS1/0 on input line 390.
Package textcomp Info: Setting qtm sub-encoding to TS1/0 on input line 391.
Package textcomp Info: Setting qzc sub-encoding to TS1/0 on input line 392.
Package textcomp Info: Setting qhvc sub-encoding to TS1/0 on input line 393.
Package textcomp Info: Setting futs sub-encoding to TS1/4 on input line 394.
Package textcomp Info: Setting futx sub-encoding to TS1/4 on input line 395.
Package textcomp Info: Setting futj sub-encoding to TS1/4 on input line 396.
Package textcomp Info: Setting hlh sub-encoding to TS1/3 on input line 397.
Package textcomp Info: Setting hls sub-encoding to TS1/3 on input line 398.
Package textcomp Info: Setting hlst sub-encoding to TS1/3 on input line 399.
Package textcomp Info: Setting hlct sub-encoding to TS1/5 on input line 400.
Package textcomp Info: Setting hlx sub-encoding to TS1/5 on input line 401.
Package textcomp Info: Setting hlce sub-encoding to TS1/5 on input line 402.
Package textcomp Info: Setting hlcn sub-encoding to TS1/5 on input line 403.
Package textcomp Info: Setting hlcw sub-encoding to TS1/5 on input line 404.
Package textcomp Info: Setting hlcf sub-encoding to TS1/5 on input line 405.
Package textcomp Info: Setting pplx sub-encoding to TS1/3 on input line 406.
Package textcomp Info: Setting pplj sub-encoding to TS1/3 on input line 407.
Package textcomp Info: Setting ptmx sub-encoding to TS1/4 on input line 408.
Package textcomp Info: Setting ptmj sub-encoding to TS1/4 on input line 409.
))
(c:/Users/monik/AppData/Roaming/TinyTeX/texmf-dist/tex/latex/microtype/microtyp
e.sty
Package: microtype 2019/02/28 v2.7b Micro-typographical refinements (RS)
\MT@toks=\toks22
\MT@count=\count273
LaTeX Info: Redefining \textls on input line 790.
\MT@outer@kern=\dimen147
LaTeX Info: Redefining \textmicrotypecontext on input line 1336.
\MT@listname@count=\count274
(c:/Users/monik/AppData/Roaming/TinyTeX/texmf-dist/tex/latex/microtype/microtyp
e-xetex.def
File: microtype-xetex.def 2019/02/28 v2.7b Definitions specific to xetex (RS)
LaTeX Info: Redefining \lsstyle on input line 258.
)
Package microtype Info: Loading configuration file microtype.cfg.
(c:/Users/monik/AppData/Roaming/TinyTeX/texmf-dist/tex/latex/microtype/microtyp
e.cfg
File: microtype.cfg 2019/02/28 v2.7b microtype main configuration file (RS)
))
(c:/Users/monik/AppData/Roaming/TinyTeX/texmf-dist/tex/latex/geometry/geometry.
sty
Package: geometry 2018/04/16 v5.8 Page Geometry
(c:/Users/monik/AppData/Roaming/TinyTeX/texmf-dist/tex/generic/oberdiek/ifpdf.s
ty
Package: ifpdf 2018/09/07 v3.3 Provides the ifpdf switch
)
(c:/Users/monik/AppData/Roaming/TinyTeX/texmf-dist/tex/generic/oberdiek/ifvtex.
sty
Package: ifvtex 2016/05/16 v1.6 Detect VTeX and its facilities (HO)
Package ifvtex Info: VTeX not detected.
)
\Gm@cnth=\count275
\Gm@cntv=\count276
\c@Gm@tempcnt=\count277
\Gm@bindingoffset=\dimen148
\Gm@wd@mp=\dimen149
\Gm@odd@mp=\dimen150
\Gm@even@mp=\dimen151
\Gm@layoutwidth=\dimen152
\Gm@layoutheight=\dimen153
\Gm@layouthoffset=\dimen154
\Gm@layoutvoffset=\dimen155
\Gm@dimlist=\toks23
)