; Alife Project ; Ken Webb ; (C) Copyright 2003 Ken Webb ; All rigghts reserved ; December 18, 2003 ; Sopc_CholSopc_4.nlogo ; Proteins breeds [ proteins ] ; Lipid variables patches-own [ lipidType lipidCurvature lipidThickness ] ; Protein variables proteins-own [ proteinL ; length of protein hydrophobic length (Lundbaek p.2081, 2082) ] ; Global Constants and Variables globals [ ; Types of lipids sopcType ; 1-stearoyl-2-oleoyl-phosphatidylcholine (SOPC) (100%) cholSopcType ; Cholesterol and Sopc together (1:1 50% each) ; Thicknesses of the lipid types in nanometers (nm) sopcThickness ; Sopc lipids cholSopcThickness ; CholSopc lipids meanThickness ; Mean thickness of bilayer when Sopc:CholSopc is 1:1 meanL ; mean protein TMD length ; Colors used to display the lipid types sopcColor ; Sopc cholSopcColor ; CholSopc noColor ; No color thicknessColor ; Base color to use when coloring by thickness only proteinColor ; Base color to use when coloring proteins ; Constant for converting number of amino acids (AA) in protein TMD, to nm convertAA2nm ; Indices into list used in swap routine LTypeIx LThickIx LColIx ; Global flags OldLipidColoringScheme ; Misc. tempTerm ; term to add to proteinL to get a value within proteinThickness range plotInterval ; only refresh plot every plotInterval time steps globalValList ; used for swap generationCount ; shows passage of time (time steps) budInterval ; try budding every budInterval time steps bud_Radius ; radius of the bud region budTotalLipidsInRadius ; total number of lipids within random radius budCompareVal ; number if n% are cholSopcType chromosome ; list of lists of possible Protein & Lipid movement probability values ; Probability matrix of protein (P_) movement ; see seek-optimal-site function ; Gt = greater than ; Eq = equal ; Lt = less than P_ProbGtGt P_ProbGtEq P_ProbGtLt P_ProbEqGt P_ProbEqEq P_ProbEqLt P_ProbLtGt P_ProbLtEq P_ProbLtLt ; Probability matrix of lipid (L_) movement L_ProbIf0 L_ProbIf1 L_ProbIf2 L_ProbIf3 L_ProbIf4 L_ProbIf5 L_ProbIf6 L_ProbIf7 L_ProbIf8 ] ; setup everything including the lipid bilayer and proteins to setup ca clear-output set-globals set-probabilities ChromosomeNum chromosome set budTotalLipidsInRadius value-from (patch 0 0) [count patches in-radius bud_Radius] set budCompareVal budTotalLipidsInRadius * 0.5 ; bud when 50% are cholSopcType setup-lipidConfigurations ask patches [ color-lipids ] report-colors initialize-proteins ; histogram plot setup-optimality-plot plot-optimality plot-sorting end ; Setup plot to setup-optimality-plot set-current-plot "Level of Optimality" set-plot-x-range -1.0 1.0 set-plot-y-range 0 count proteins set-histogram-num-bars 21 end ; Plot Histogram to plot-optimality set-current-plot "Level of Optimality" histogram proteins [proteinL - lipidThickness] end ; Plot Sorting to plot-sorting locals [tmd15 tmd16 tmd17 tmd18 tmd19 tmd20] if count proteins < 100 [stop] ; prevent a divide-by-zero error set-current-plot "Proteins in CholSopc Lipid" ; TMD 15 amino acids set-current-plot-pen "Tmd15" set tmd15 count proteins with [proteinL = 2.85 and lipidType = cholSopcType] / count proteins with [proteinL = 2.85] * 100 plot tmd15 ; TMD 16 amino acids set-current-plot-pen "Tmd16" set tmd16 count proteins with [proteinL = 3.0 and lipidType = cholSopcType] / count proteins with [proteinL = 3.0] * 100 plot tmd16 ; TMD 17 amino acids set-current-plot-pen "Tmd17" set tmd17 count proteins with [proteinL = 3.15 and lipidType = cholSopcType] / count proteins with [proteinL = 3.15] * 100 plot tmd17 ; TMD 18 amino acids set-current-plot-pen "Tmd18" set tmd18 count proteins with [proteinL = 3.3 and lipidType = cholSopcType] / count proteins with [proteinL = 3.3] * 100 plot tmd18 ; TMD 19 amino acids set-current-plot-pen "Tmd19" set tmd19 count proteins with [proteinL = 3.45 and lipidType = cholSopcType] / count proteins with [proteinL = 3.45] * 100 plot tmd19 ; TMD 20 amino acids set-current-plot-pen "Tmd20" set tmd20 count proteins with [proteinL = 3.6 and lipidType = cholSopcType] / count proteins with [proteinL = 3.6] * 100 plot tmd20 end ; Bud and Dock to bud-and-dock locals [x y r lCount pCount] ; create circular regions at random locations set x (random screen-size-x) - screen-edge-x set y (random screen-size-y) - screen-edge-y set r bud_Radius ; not sure what this should be yet ;ask (patch x y) [show count (patches in-radius r) with [lipidType = cholSopcType]] set lCount value-from (patch x y) [count (patches in-radius r) with [lipidType = cholSopcType]] ;show lCount if lCount > budCompareVal [ ; Bud lipids ask (patch x y) [ask (patches in-radius r) [set-all-lipid-variables sopcType sopcThickness lime]] ; Bud and Dock proteins - by random repositioning, rather than die and recreate ;set pCount value-from (patch x y) [count (proteins in-radius r)] ask (patch x y) [ask (proteins in-radius r) [setxy ((random screen-size-x) - screen-edge-x) ((random screen-size-y) - screen-edge-y)]] ; Dock lipids - fill in same number of random locations while [lCount > 0] [ set x (random screen-size-x) - screen-edge-x set y (random screen-size-y) - screen-edge-y if value-from (patch x y) [lipidType] = sopcType [ ask (patch x y) [set-all-lipid-variables cholSopcType cholSopcThickness yellow] ] set lCount lCount - 1 ] ; end while ] ; end if end ; Go to go if LipidColoringScheme != OldLipidColoringScheme [ ask patches [ color-lipids ] set OldLipidColoringScheme LipidColoringScheme ] ask proteins [seek-optimal-site] if InitialLipidConfiguration = "RandomOnce" or InitialLipidConfiguration = "RandomOnceCircles" [ ask patches [adjust-domain-size-continuous] ] if generationCount mod plotInterval = 0 [plot-optimality] if generationCount mod budInterval = 0 [bud-and-dock] plot-sorting set generationCount generationCount + 1 end ; Seek optimal site depending on TMD length ; The protein has two questions: ; 1. What state am I currently in? (compare myVal with 0) ; 2. What state would I be in if I moved? (compare bnVal with myVal) ; to seek-optimal-site ; protein procedure locals [pL bestNeighbor bnVal myVal x y] set pL proteinL set bestNeighbor random-one-of neighbors set bnVal value-from bestNeighbor [pL - lipidThickness] set myVal pL - lipidThickness ; value at current location set x value-from bestNeighbor [pxcor] set y value-from bestNeighbor [pycor] if myVal > 0 ; TMD partly exposed to water [ if bnVal > myVal and random P_ProbGtGt = 1 [setxy x y] ; get worse if bnVal = myVal and random P_ProbGtEq = 1 [setxy x y] ; stay the same if bnVal < myVal and random P_ProbGtLt = 1 [setxy x y] ; get better ] if MyVal = 0 ; TMD optimal [ if bnVal > myVal and random P_ProbEqGt = 1 [setxy x y] ; get much worse if bnVal = myVal and random P_ProbEqEq = 1 [setxy x y] ; stay optimal if bnVal < myVal and random P_ProbEqLt = 1 [setxy x y] ; get somewhat worse ] if myVal < 0 ; TMD embedded in lipid with room to spare [ if bnVal > myVal and random P_ProbLtGt = 1 [setxy x y] ; get better if bnVal = myVal and random P_ProbLtEq = 1 [setxy x y] ; stay the same if bnVal < myVal and random P_ProbLtLt = 1 [setxy x y] ; get somewhat worse ] end ; Give values to all the global variables to set-globals ; Constants set sopcType 1 ; type set cholSopcType 2 set sopcThickness 3.0 ; thickness in nm set cholSopcThickness 3.3 set sopcColor green ; color set cholSopcColor red set noColor white set thicknessColor orange set proteinColor blue set convertAA2nm 20 / 3 ; about 6.667 set tempTerm 0.6 ; not sure if I need this or not, so it's temporary set plotInterval 5 ; plot every plotInterval time steps ; Bud set budInterval 1 ; try budding every budInterval time steps set bud_Radius 7 ; default ; end Bud set LTypeIx 0 ; list indices used in swap routine set LThickIx 1 set LColIx 2 ; Variables set meanThickness mean list sopcThickness cholSopcThickness set OldLipidColoringScheme LipidColoringScheme set globalValList [] set generationCount 0 ; Probability matrix for Protein (P_) movement (values in range from 1 to 16). ; prob = 1 is 100% certain ; prob = 16 is highly unlikely ; default 8 3 2 12 3 4 2 3 4 ; perfect sorting 100000 3 2 100000 3 100000 2 3 100000 set chromosome [ [1 [ 8 3 2 12 3 4 2 3 4 2 16 64 160 320 640 10000 100000 1000000 99]] ; default [2 [100000 3 2 100000 3 100000 2 3 100000 1 2 4 5 10000 10000 10000 100000 1000000 99]] ; perfect sorting ] end ; Set the probability variables from values in chromosome list to set-probabilities [probID pList] ifelse empty? pList [print "Can't find pList"] [ifelse first first pList = probID [set-prob-detail 1 first but-first first pList] [set-probabilities probID but-first pList] ] end to set-prob-detail [itemID pList] ; print pList ; Protein movement (P_) probabilities if itemID = 1 [set P_ProbGtGt first pList] if itemID = 2 [set P_ProbGtEq first pList] if itemID = 3 [set P_ProbGtLt first pList] if itemID = 4 [set P_ProbEqGt first pList] if itemID = 5 [set P_ProbEqEq first pList] if itemID = 6 [set P_ProbEqLt first pList] if itemID = 7 [set P_ProbLtGt first pList] if itemID = 8 [set P_ProbLtEq first pList] if itemID = 9 [set P_ProbLtLt first pList] ; Lipid movement (L_) probabilities if itemID = 10 [set L_ProbIf0 first pList] if itemID = 11 [set L_ProbIf1 first pList] if itemID = 12 [set L_ProbIf2 first pList] if itemID = 13 [set L_ProbIf3 first pList] if itemID = 14 [set L_ProbIf4 first pList] if itemID = 15 [set L_ProbIf5 first pList] if itemID = 16 [set L_ProbIf6 first pList] if itemID = 17 [set L_ProbIf7 first pList] if itemID = 18 [set L_ProbIf8 first pList] ; Bud Radius if itemID = 19 [ set bud_Radius first pList if bud_Radius = 99 [set bud_Radius BudRadius] ; allow it to be set from GUI ] if itemID < 19 [set-prob-detail itemID + 1 but-first pList] end ; Setup initial lipid configuration to setup-lipidConfigurations ; observer procedure if InitialLipidConfiguration = "Random" [ initialize-lipidBilayer-Random set-lipid-gradient 1 ] if InitialLipidConfiguration = "RandomOnce" [ initialize-lipidBilayer-RandomOnce set-lipid-gradient 1 ] if InitialLipidConfiguration = "RandomOnceCircles" [ initialize-lipidBilayer-RandomOnceCircles set-lipid-gradient 1 ] if InitialLipidConfiguration = "ConstantGradientRectangular" [ initialize-lipidBilayer-ConstantGradientRectangle set-lipid-gradient 100 ] end ; Initialize the lipid bilayer to initialize-lipidBilayer-Random ; observer procedure locals [loopCounter] ; to prevent an infinite loop set loopCounter 200 ask patches [set-lipid-type-Random] while [domain-size-ok? = false and loopCounter > 0] [ ;type count patches with [lipidType = cholSopcType] type " " ask patches [adjust-domain-size] set loopCounter loopCounter - 1 ] ask patches [set-lipid-thickness] end ; Initialize the lipid bilayer to initialize-lipidBilayer-RandomOnce ; observer procedure ask patches [set-lipid-type-Random] ;ask (patch 0 0) [ask patches in-radius 10 [set lipidType cholSopcType] ] ask patches [set-lipid-thickness] end ; Initialize the lipid bilayer to initialize-lipidBilayer-RandomOnceCircles ; observer procedure locals [x y r totalPatches cholSopcPatches percentChol] ;ask patches [set-lipid-type-Random] ask patches [set lipidType sopcType] set totalPatches count patches set percentChol PercentCholesterolSopc / 100 set cholSopcPatches count patches with [lipidType = cholSopcType] ;repeat 250 [ while [(cholSopcPatches / totalPatches) < percentChol] [ set x (random screen-size-x) - screen-edge-x set y (random screen-size-y) - screen-edge-y ;set r random 25 ;set r random-normal 10 5 ;set r random-exponential 2 set r random-poisson 1 ask (patch x y) [ask patches in-radius r [set lipidType cholSopcType] ] set cholSopcPatches count patches with [lipidType = cholSopcType] ] ask patches [set-lipid-thickness] end ; Initialize the lipid bilayer to initialize-lipidBilayer-ConstantGradientRectangle ; observer procedure ask patches [set-lipid-type-ConstantGradientRectangle] ask patches [set-lipid-thickness] end ; Set lipid gradient (recursive function) to set-lipid-gradient [n] ; observer procedure if n > 0 [ diffuse lipidThickness DiffusionConstant ; smooth the distribution of thicknesses set-lipid-gradient n - 1 ] end ; Initialize proteins to initialize-proteins ; observer procedure set-default-shape proteins "circle" create-custom-proteins NumberOfProteins [ setxy (random screen-size-x) (random screen-size-y) ; to get a range from 2.25 to 3.00 nm (15 to 20 amino acids in TMD) set proteinL (random 6 + 15) / convertAA2nm + tempTerm ] set meanL mean values-from proteins [proteinL] ask proteins [set color proteinColor - ((proteinL - meanL) * 10)] end ; Set lipid type to set-lipid-type-Random ; patches procedure ifelse random 100 < PercentCholesterolSopc [set lipidType cholSopcType] ; CholSopc [set lipidType sopcType] ; SOPC end ; Set lipid type to set-lipid-type-ConstantGradientRectangle ; patches procedure ifelse pxcor > 0 ; if the patch is right of the center line [set lipidType cholSopcType] ; CholSopc [set lipidType sopcType] ; SOPC end ; Adjust domain size to adjust-domain-size ; patches procedure if lipidType = sopcType [ if count neighbors with [lipidType = sopcType] < 2 [swap-lipids cholSopcType] ] if lipidType = cholSopcType [ if count neighbors with [lipidType = cholSopcType] < MinNeighborhoodSize [swap-lipids sopcType] ] end ; Adjust domain size (continuously) to adjust-domain-size-continuous ; patches procedure locals [neighCount] if lipidType = sopcType [ if count neighbors with [lipidType = sopcType] < 2 [swap-lipids cholSopcType] ] if lipidType = cholSopcType [ set neighCount count neighbors with [lipidType = cholSopcType] if neighCount = 0 and random L_ProbIf0 = 0 [swap-lipids sopcType] if neighCount = 1 and random L_ProbIf1 = 0 [swap-lipids sopcType] if neighCount = 2 and random L_ProbIf2 = 0 [swap-lipids sopcType] if neighCount = 3 and random L_ProbIf3 = 0 [swap-lipids sopcType] if neighCount = 4 and random L_ProbIf4 = 0 [swap-lipids sopcType] if neighCount = 5 and random L_ProbIf5 = 0 [swap-lipids sopcType] if neighCount = 6 and random L_ProbIf6 = 0 [swap-lipids sopcType] if neighCount = 7 and random L_ProbIf7 = 0 [swap-lipids sopcType] if neighCount = 8 and random L_ProbIf8 = 0 [swap-lipids sopcType] ] end ; Swap lipids to swap-lipids [swapeeType] ; patches procedure locals [lType lThick lCol valList somebody] set lType lipidType ;set lCurv lipidCurvature set lThick lipidThickness set lCol pcolor without-interruption [ set somebody random-one-of neighbors with [lipidType = swapeeType] if somebody != nobody [ask somebody [swap-all-lipid-variables lType lThick lCol]] set valList globalValList ] set lipidType item LTypeIx valList ;set lipidCurvature item LCurvIx valList set lipidThickness item LThickIx valList set pcolor item LColIx valList end ; Swap all lipid variables to swap-all-lipid-variables [lType lThick lCol] ; patches procedure locals [oldVals] set oldVals (list lipidType lipidThickness pcolor) set-all-lipid-variables lType lThick lCol ;set lipidType lType ;set lipidThickness lThick ;set pcolor lCol set globalValList oldVals end ; Set all lipid variables to set-all-lipid-variables [lType lThick lCol] ; patches procedure set lipidType lType set lipidThickness lThick set pcolor lCol end ; Is the domain size OK for all patches ? to-report domain-size-ok? ; observer procedure ifelse count patches with [lipidType = cholSopcType and count neighbors with [lipidType = cholSopcType] < MinNeighborhoodSize ] = 0 [report true] [report false] end ; Set default lipid thickness to set-lipid-thickness ; patches procedure ifelse lipidType = sopcType [ set lipidThickness sopcThickness ] [ set lipidThickness cholSopcThickness ] end ; Update lipid coloring scheme from GUI button to update if LipidColoringScheme != OldLipidColoringScheme [ ask patches [ color-lipids ] set OldLipidColoringScheme LipidColoringScheme ] end ; Color lipids based on settings of GUI choosers to color-lipids ; patches procedure if LipidColoringScheme = "ColorByLipidType" [color-lipid-type] if LipidColoringScheme = "ColorByLipidThickness" [color-lipid-thickness] if LipidColoringScheme = "ColorByBoth" [color-lipid-typeAndThickness] if LipidColoringScheme = "ColorByNone" [color-lipid-none] end ; Color lipids based on their type only to color-lipid-type ; patches procedure ifelse lipidType = sopcType [ set pcolor sopcColor ] [ set pcolor cholSopcColor ] end ; color lipids based on their thickness only to color-lipid-thickness ; patches procedure set pcolor thicknessColor - ((lipidThickness - meanThickness) * 10) end ; Color lipids based on both their type and their thickness to color-lipid-typeAndThickness ; patches procedure color-lipid-type diffuse-colors end ; Color lipids using neither type nor thickness to color-lipid-none ; patches procedure set pcolor noColor end ; Diffuse colors based on lipid thickness to diffuse-colors ; patches procedure ifelse lipidType = sopcType [ set pcolor pcolor - ((lipidThickness - sopcThickness) * 10) ] [ set pcolor pcolor - ((lipidThickness - cholSopcThickness) * 10) ] end ; Report the meaning of the different colors in the model to report-colors type "Sopc lipid is " report-color sopcColor type ". CholSopc combination is " report-color cholSopcColor print "." print "For protein TMD colors, see legend in graph at left." end to report-color [lColor] locals [strColor] set strColor lColor if shade-of? red lColor [set strColor "red"] if shade-of? green lColor [set strColor "green"] if shade-of? blue lColor [set strColor "blue"] type strColor end @#$#@#$#@ GRAPHICS-WINDOW 321 10 733 443 100 100 2.0 0 10 0 0 CC-WINDOW 321 453 734 572 Command Center BUTTON 231 62 314 95 NIL setup NIL 1 T OBSERVER SLIDER 7 10 205 43 PercentCholesterolSopc PercentCholesterolSopc 0 100 50 1 1 NIL MONITOR 7 472 179 521 AverageBilayerThickness sum values-from patches [lipidThickness] / count patches 3 1 SLIDER 7 98 205 131 NumberOfProteins NumberOfProteins 10 5000 5000 10 1 NIL BUTTON 231 99 314 132 NIL go T 1 T OBSERVER CHOICE 7 144 202 189 LipidColoringScheme LipidColoringScheme "ColorByLipidType" "ColorByLipidThickness" "ColorByBoth" "ColorByNone" 0 SLIDER 744 309 921 342 MinNeighborhoodSize MinNeighborhoodSize 0 4 3 1 1 NIL BUTTON 202 149 280 182 NIL update NIL 1 T OBSERVER CHOICE 7 48 205 93 InitialLipidConfiguration InitialLipidConfiguration "Random" "RandomOnce" "RandomOnceCircles" "ConstantGradientRectangular" 1 SLIDER 743 229 915 262 DiffusionConstant DiffusionConstant 0.0 1.0 0.0 0.1 1 NIL PLOT 743 10 1048 223 Level of Optimality Protein Optimality Quantity of Proteins -1.0 1.0 0.0 2000.0 false false PENS "proteins" 1.0 1 -8716033 true SLIDER 7 199 145 232 ChromosomeNum ChromosomeNum 1 2 2 1 1 NIL MONITOR 231 10 314 59 Time Step generationCount 0 1 PLOT 7 244 293 467 Proteins in CholSopc Lipid Time Step Concentration (% in rafts) 0.0 100.0 0.0 100.0 true true PENS "Tmd15" 1.0 0 -4210689 true "Tmd16" 1.0 0 -9210881 true "Tmd17" 1.0 0 -14276865 true "Tmd18" 1.0 0 -16776999 true "Tmd19" 1.0 0 -16777076 true "Tmd20" 1.0 0 -16777152 true TEXTBOX 744 342 1036 360 Only useful when InitialLipidConfiguration = Random. TEXTBOX 743 262 988 297 Only useful when InitialLipidConfiguration = Random or ConstantGradientRectangular. SLIDER 150 199 314 232 BudRadius BudRadius 1 20 10 1 1 NIL @#$#@#$#@ WHAT IS IT? ----------- This section could give a general understanding of what the model is trying to show or explain. There are two types of lipids in the model: 1 1-stearoyl-2-oleoyl-phosphatidylcholine (SOPC) 2 Cholesterol Each location on the 2D grid is assumed to contain exactly one pair of lipids with their tails facing each other. HOW IT WORKS ------------ The lipid bilayer is not explicitly modeled. Properties such as the average bilayer thickness emerge out of the local interactions of the individual lipid molecules. The average bilayer thickness is a dependent variable, that depends on the independent variable PercentCholesterol. HOW TO USE IT ------------- Set the PercentCholesterol slider to a value between 0% and 100% cholesterol. The Golgi bilayer is about 50% cholesterol. Press the setup button. THINGS TO NOTICE ---------------- SOPC lipids are shown in green, cholesterol lipids in red. The thicker either type of lipid is, the darker the color. LipidType LipidThickness --------- -------------- 1 SOPC 3.0 nm 2 Cholesterol 3.6 nm (KSW assumption) These are the default thicknesses for each lipid type, when lipid molecules are surrounded by a fairly large number of other molecules of the same type. There are two resulting regions of lipid bilayer: BilayerType Bilayer hydrophobic thickness d0 ------------------- -------------------------------- 1 SOPC Bilayer 3.0 nm (approx.) 2 SOPC:Chol Bilayer (50% cholesterol) 3.3 nm (approx.) "We further assume that the bilayer thickness varies as a linear function of the cholesterol mole fraction." (source: Lundbaek, 2003, p.2082) In the NetLogo model: PercentCholesterol AverageBilayerThickness ------------------ ----------------------- 0% 3.0 50% 3.3 (approx.) 100% 3.6 THINGS TO TRY ------------- This section could give some ideas of things for the user to try to do (move sliders, switches, etc.) with the model. EXTENDING THE MODEL ------------------- This section could give some ideas of things to add or change in the procedures tab to make the model more complicated, detailed, accurate, etc. NETLOGO FEATURES ---------------- This section could point out any especially interesting or unusual features of NetLogo that the model makes use of, particularly in the Procedures tab. It might also point out places where workarounds were needed because of missing features. RELATED MODELS -------------- This section could give the names of models in the NetLogo Models Library or elsewhere which are of related interest. CREDITS AND REFERENCES ---------------------- This section could contain a reference to the model's URL on the web if it has one, as well as any other necessary credits or references. Copyright (C) 2003 by Ken Webb. All rights reserved. @#$#@#$#@ default true 0 Polygon -7566196 true true 150 5 40 250 150 205 260 250 ant true 0 Polygon -7566196 true true 136 61 129 46 144 30 119 45 124 60 114 82 97 37 132 10 93 36 111 84 127 105 172 105 189 84 208 35 171 11 202 35 204 37 186 82 177 60 180 44 159 32 170 44 165 60 Polygon -7566196 true true 150 95 135 103 139 117 125 149 137 180 135 196 150 204 166 195 161 180 174 150 158 116 164 102 Polygon -7566196 true true 149 186 128 197 114 232 134 270 149 282 166 270 185 232 171 195 149 186 149 186 Polygon -7566196 true true 225 66 230 107 159 122 161 127 234 111 236 106 Polygon -7566196 true true 78 58 99 116 139 123 137 128 95 119 Polygon -7566196 true true 48 103 90 147 129 147 130 151 86 151 Polygon -7566196 true true 65 224 92 171 134 160 135 164 95 175 Polygon -7566196 true true 235 222 210 170 163 162 161 166 208 174 Polygon -7566196 true true 249 107 211 147 168 147 168 150 213 150 arrow true 0 Polygon -7566196 true true 150 0 0 150 105 150 105 293 195 293 195 150 300 150 bee true 0 Polygon -256 true false 151 152 137 77 105 67 89 67 66 74 48 85 36 100 24 116 14 134 0 151 15 167 22 182 40 206 58 220 82 226 105 226 134 222 Polygon -16777216 true false 151 150 149 128 149 114 155 98 178 80 197 80 217 81 233 95 242 117 246 141 247 151 245 177 234 195 218 207 206 211 184 211 161 204 151 189 148 171 Polygon -7566196 true true 246 151 241 119 240 96 250 81 261 78 275 87 282 103 277 115 287 121 299 150 286 180 277 189 283 197 281 210 270 222 256 222 243 212 242 192 Polygon -16777216 true false 115 70 129 74 128 223 114 224 Polygon -16777216 true false 89 67 74 71 74 224 89 225 89 67 Polygon -16777216 true false 43 91 31 106 31 195 45 211 Line -1 false 200 144 213 70 Line -1 false 213 70 213 45 Line -1 false 214 45 203 26 Line -1 false 204 26 185 22 Line -1 false 185 22 170 25 Line -1 false 169 26 159 37 Line -1 false 159 37 156 55 Line -1 false 157 55 199 143 Line -1 false 200 141 162 227 Line -1 false 162 227 163 241 Line -1 false 163 241 171 249 Line -1 false 171 249 190 254 Line -1 false 192 253 203 248 Line -1 false 205 249 218 235 Line -1 false 218 235 200 144 bird1 false 0 Polygon -7566196 true true 2 6 2 39 270 298 297 298 299 271 187 160 279 75 276 22 100 67 31 0 bird2 false 0 Polygon -7566196 true true 2 4 33 4 298 270 298 298 272 298 155 184 117 289 61 295 61 105 0 43 boat1 false 0 Polygon -1 true false 63 162 90 207 223 207 290 162 Rectangle -6524078 true false 150 32 157 162 Polygon -16776961 true false 150 34 131 49 145 47 147 48 149 49 Polygon -7566196 true true 158 33 230 157 182 150 169 151 157 156 Polygon -7566196 true true 149 55 88 143 103 139 111 136 117 139 126 145 130 147 139 147 146 146 149 55 boat2 false 0 Polygon -1 true false 63 162 90 207 223 207 290 162 Rectangle -6524078 true false 150 32 157 162 Polygon -16776961 true false 150 34 131 49 145 47 147 48 149 49 Polygon -7566196 true true 157 54 175 79 174 96 185 102 178 112 194 124 196 131 190 139 192 146 211 151 216 154 157 154 Polygon -7566196 true true 150 74 146 91 139 99 143 114 141 123 137 126 131 129 132 139 142 136 126 142 119 147 148 147 boat3 false 0 Polygon -1 true false 63 162 90 207 223 207 290 162 Rectangle -6524078 true false 150 32 157 162 Polygon -16776961 true false 150 34 131 49 145 47 147 48 149 49 Polygon -7566196 true true 158 37 172 45 188 59 202 79 217 109 220 130 218 147 204 156 158 156 161 142 170 123 170 102 169 88 165 62 Polygon -7566196 true true 149 66 142 78 139 96 141 111 146 139 148 147 110 147 113 131 118 106 126 71 box true 0 Polygon -7566196 true true 45 255 255 255 255 45 45 45 butterfly1 true 0 Polygon -16777216 true false 151 76 138 91 138 284 150 296 162 286 162 91 Polygon -7566196 true true 164 106 184 79 205 61 236 48 259 53 279 86 287 119 289 158 278 177 256 182 164 181 Polygon -7566196 true true 136 110 119 82 110 71 85 61 59 48 36 56 17 88 6 115 2 147 15 178 134 178 Polygon -7566196 true true 46 181 28 227 50 255 77 273 112 283 135 274 135 180 Polygon -7566196 true true 165 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