UBC Theses and Dissertations

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UBC Theses and Dissertations

Physiological effects of radiant energy Ashton, Emily Miriam Ruth 1932

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U ! B . C . LIBRARY A(£. NO. mBioioaxcAL  wwmm  OF RAPIAIT  B i l l y Miriam Rath Ashton.  A ffeesis s u b m i t t e d f o r t h e degree o f  l a the  M P A l f s I l i f OF -BOf A l l .  the Snipers it y of B r i t i s h G e l a a l i a April*  1932.  7f ^7 c  0  0 ft f  R  I f S.  Introduction  ....  Aekn owled gment s  Page  ...........  1 2  Experimental f o r k - Part A. I n t r o d u c t i o n and Review of l i t e r a t u r e ....  3  Experimental WorkPart 1.......  8  Part 2.......  14  Comparison o f Results  21  Soanary of Results  24  E x p e r i m e n t a l Work- Part 3. Introd uct ion  ....  Review o f L i t e r a t u r e  £6 28  Experimental f o r k Part 1. .....  29  Part 2. .....  43  Snsmary ©f Results  61  L i t e r a t u r e c i t e d ...........  53  H H 3 I 0 L 0 6 I C A L EFFECTS OF RAPIAHf ENERGY  INTRQPOeflOH.  Since the  very e a r l i e s t t i n e s l i g h t has  worshipped as t h e g i v e r o f h e a l t h and o f death, but  darkness and recent  enemy  of  i t i s o n l y i n comparatively  y e a r s t h a t s c i e n t i f i c e n q n i r y has  as t o i t s a c t u a l b i o l o g i c a l and The  as the  been  been c a r r i e s ' on  physiological effects.  amount of l i t e r a t u r e on t h e subject i s immense and  v a r i e d , so t h a t i t i s i m p o s s i b l e account o f any  bat the  to p i v e a comprehensive  r e s u l t s 010st p e r t i n e n t t o  the  work described i n t h i s d i s c u s s i o n . For  the  p a s t f o u r y e a r s work on t h e e f f e c t  of  R a d i a n t Energy on P r o t o p l a s m has been c a r r i e d on under  the  d i r e c t i o n of Dr. Hutchinson and  the  i n o r d e r to understand  r e a s o n s l e a d i n g up to the i n v e s t i g a t i o n o f ensyae a c t i o n oatlined  i n the l a t t e r p o r t i o n of t h i s paper, i t i s n e c -  essary to d e s c r i b e the I t was t h i s paper i n t o two  p r e l i m i n a r y work.  t h e r e f o r e thought a d v i s a b l e t o d i v i d e p a r t s , A and  B; h to consist of a  d e s c r i p t i o n o f t h e work on P r o t o p l a s m and  3 to c o n s i s t of a  description of the effect of R a d i a n t Rnerpy on ensyae action i n p a r t i e a l a r .  ACMOlOTKMlSfS G r a t e f u l acknowledgment i s mace t o D r . A. H» H u t c h i n s o n , under whose d i r e c t i o n t h i s work has been done, f o r h i s u n f a i l i n g h e l p and encouragement. f h e a u t h o r i s a l s o i n d e b t e d t o the N a t i o n a l R e s e a r c h C o u n c i l o f Canada f o r t h e nee o f l i g h t s o u r c e s and  instruaents i n their possession.  3 PARS A. f E l EFFECT OF RABIAIf KfTEROY OH PROTOPLASM. IHTROB0CTIOH.  Man's c o n t r o l of the e f f e c t s of l i g h t and more e s p e c i a l l y u l t r a v i o l e t l i g h t i s at present incomplete and although the e f f e c t s nay d i f f e r w i t h wave l e n g t h and i n t e n s i t y and w i t h the subject, i r r a d i a t e d l i t t l e I s known a© to the why or the wherefore o f the r e s u l t s obtained. I t was thought that a comparison between the e f f e c t o f Radiant Energy and more p a r t i c u l a r l y Monochromatic Light on animal and plant protoplasm might help towards the understanding of these phenomena and towards t h e i r more h e l p f u l u t i l i s a t i o n .  REVIEW OF LITERATURE. 1.  E f f e c t of Light on Animal Protoplasm* L i g h t can have no e f f e c t unless I t i s absorbed  and according t o Boyle (3) "rays a f f e c t protoplasm at the place where they are absorbed, and observed  physiological  disturbances a r e the responses on the part of the organism to i t s injured protoplasm". statement Bovie  In a m p l i f i c a t i o n of t h i s  and h i s co-workers ( 4 ) have demonstrated  the e f f e c t of various l i g h t rays on Protoplasm. moeoiuos c a n i a t n a was exposed  Para-  to u l t r a v i o l e t l i g h t i n a  4  f l u e r i t e chamber and the organism was rendered extremely s e n s i t i v e t o heat, so much so that i t was k i l l e d  by an  amount o f heat which would not a f f e c t i t normally.  The  Sehoaaan r e g i o n (5) i s a region o f general absorption f o r n e a r l y a l l substances and has an immediate e f f e c t on i n f u s o r i a , causing marked s t i m u l a t i o n followed by c y t o l y s i s and With s u f f i c i e n t exposure, death-  Packard (25) f i n d s that  Paramecium i s s u s c e p t i b l e t o Badina r a d i a t i o n s and t h a t the s u s c e p t i b i l i t y to these r a d i a t i o n s v a r i e s w i t h the p e r m e a b i l i t y of the surface l a y e r o f the c e l l .  He suggests  that the increased p e r m e a b i l i t y i s the cause o f the a c c e l e r a t i o n i n the d i v i s i o n r a t e observed.  Hutchinson and Ashton  (18) found that moderate increase i n r a t e of p l a s n o l y s i s accompanied increase In c e l l d i v i s i o n i n the same organism and that an excessive Increase  i n r a t e of p l a s a o l y s i s accom-  panied decrease i n growth r a t e , but whether t h i s change i n plasmolytic r a t e was due t o a changed p e r m e a b i l i t y or to the change i n the osmotic content of the c e l l has not been determined. According  to Hughes and Bovie (16) c y t o l y s i s i n  Paraaoeciura can be caused by exposure to u l t r a v i o l e t l i g h t , an exposure s u f f i c i e n t t o division.  o&nae  cytolysis inhibiting c e l l  With short exposures the i n h i b i t i o n i s t r a n s i t o r y  and i s followed by an increase i n c e l l d i v i s i o n over that ©f the c o n t r o l .  5  *  &. L i g h t responses - A n i m a l . f o r k on m u l t i c e l l u l n r organisms shows t h a t r a d i a n t energy has a pr of o and  e f f e c t on growth and  Loeb (23) found t h a t u n f e r t i l i s e d  eggs of A r b a c i a , when t r e a t e d  w i t h r a y s from a q u a r t s mercury lamp formed a membrane and  development*  fertilisation  i n some oases developed i n t o l a r v a e , though not  p r o c e e d i n g beyond t h e g a s t r a l a s t a g e -  Higgins  and  Sheard  found t h a t u l t r a v i o l e t l i g h t e f f e c t e d the development o f eggs o f Rana p i p i e n s and  t h a t the r e g i o n s of h i g h e s t  a c t i v i t y were most r e s p o n s i v e even more f a r r e a c h i n g obtained  to radiation.  effect,  i l n l l u r and  a n t a t l o n s , p r o d u c i n g v i s i b l e and  (15) the  metabolic  X - r a y s have an  Weinstein  (37)  lethal effects  as w e l l as g e n e t i c m o d i f i c a t i o n s of f r e q u e n c y o f c r o s s i n g over i n B r o s o p h i l a exposed t o t h e s e r a y s and attachment  between  genes o f d i f f e r e n t c h r o a o s o n e s , p o i n t i n g to a profound e f f e c t on the r e p r o d u c t i v e  c e l l s and  Reaction recorded.  B i g g i n s and  t h e chromosomes the as e l v e s .  t o s p e c i f i c wave l e n g t h s has a l s o been Sheard (14)  found t h a t i n the absence  of u l t r a v i o l e t l i g h t , as when b l u e or amber s c r e e n s were used, h y p e r p l a s i a of the parathyroid and  glands of chickens  resulted,  t h a t t h e best development t o o k p l a c e i n d i r e c t s u n l i g h t .  Abbot t l ) s t a t e s t h a t Formi -a d a k o t e n s i s  s p e c u l a r i s responded  t o l i g h t o f the same i n t e n s i t y I n the f o l l o w i n g o r d e r ,  yellow,  w h i t e , r e d . V i s a c h e r and Luce (36) found t h a t 6300 A°o 5450 A were most e f f e c t i v e i n c a u s i n g r e a c t i o n on the l a r v a e o of barnacles. There i s a d e c r e a s e i n e f f e c t i v e n e s s t o 4200 A  6 and  a d i s t i n c t s t i m u l a t i n g e f f e c t i n the u l t r a v i o l e t . For the cure o f r i c k e t s In animals u l t r a v i o l e t  l i g h t o f v a r i o u s wave l e n g t h s  i s efficacious.  6. B.  Manghan (£4} showed t h a t t h e wave l e n g t h s between 3132 Ao o and  2650 A  a r e most e f f e c t i v e i n t h i s r e s p e c t .  e f f e c t of wave l e n g t h s s h o r t e r t h a n 2896 A s m a l l and t h a t o f wave l e n g t h 3132 A  0  0  The c u r a t i v e  seeded t o he  I t s e l f t o he n i l .  r e s u l t o f c a r e f n l a n a l y s i s showed t h a t 2968 A  0  The  was t h e most o  e f f e c t i v e f o r the c a r e o f r i c k e t s and t h a t 3024 A  was  per-  haps a f o u r t h as p o t e n t . 3. Llftht responses - P l a n t s . P r i e s t l e y (29) has performed e x p e r i m e n t s on t h e growth o f s e e d l i n g s t h a t show t h a t b r i e f expos are t o l i g h t has v e r y marked e f f e c t s .  A l i g h t exposure o f one t o two  minutes p e r day t o a r e l a t i v e l y weak a r t i f i c i a l l i g h t w i l l remove e f f e c t i v e l y  t h e most c h a r a c t e r i s t i c  morphological  f e a t u r e s o f e t i o l a t i o n , w h i l e t h e expanded l e a f w i l l f a i l t o produce any c h l o r o p h y l l . E l o g i n s and Sheard (14) found t h a t t h e l e s s e r wave l e n g t h s used, 3200 A - 2700 A , 0  and  a c c e l e r a t e d the t i m e o f g e r m i n a t i o n  0  stimulated  w h i l e g r e a t e r wave  l e n g t h s i n h i b i t e d g e r m i n a t i o n , g r o w t h b e i n g a c c e l e r a t e d by I r r a d i a t i o n i n t h e n e a r u l t r a - v i o l e t , 4000A°-3000 A ; 2900 A 0  and  0  below b e i n g d e f i n i t e l y l e t h a l . l o t a b l e r e s u l t s have been obtained  Goodspeed and 01sen ( 1 1 ) .  i n p l a n t s by  t o b a c c o i n f u l l f l o w e r was exposed  to X-rays.  P l a n t s grown from seeds I r r a d i a t e d at t h i s early  stage Ineladed many v a r i a t i o n s .  A cytologieai  investigation  showed t h a t chromosome d i s t u r b a n c e s had occurred and t h a t t h e s e mod i f l e s t ions were i n a l l p r o b a b i l i t y p r i m a r y to t h e external  variations  i n form.  Popp (£8) removed d e f i n i t e regions o f the spectrum In t h e blue-violet  and noted  a t i n g a l l l i n e s below 5890 A etiolated  appearance.  t h e e f f e c t on p l a n t s . 0  resulted  Eliminating  below 4720 A  t h e same e f f e c t , bat a f t e r e l i m i n a t i o n vary l i t t l e difference  was s e e n .  i n plants 0  Elimin-  h a v i n g an produced  of a l l ultra violet  From t h i s result  i t may  be i n f e r r e d that u l t r a v i o l e t r a y s a r e not i n d i s p e n s a b l e but t h a t t h e b i n e v i o l e t end o f t h e s p e c t r u m i s n e c c e e s a r y f o r normal v i g o r o u s g r o w t h .  8.  sxPSsx-iEHfAL IOBK. B a r t 1.  E f f e c t o f monochromatic l i g h t on ParamoecIran  f h e f i r s t p a r t o f t h e e x p e r i m e n t a l work was t o a s c e r t a i n t h e e f f e c t o f Monochromatic L i g h t on a n i m a l  proto-  plasm, P a r a a o e c l n a e a o l a t o a b e i n g t h e organise! need. f h e apparatus  used throughout  a l l the following  work was a Cooper H e w i t t '\L#*b-are" f o r t h e s o u r c e o f l i g h t . I t was operated  on an a l t e r n a t i n g o n r r e n t o f 110 v o l t s w i t h  a r e s i s t a n c e of I S ohms i n s e r i e s and fnnet ioned trader t h e s e c o n d i t i o n s as a low pressor© m e r c u r y a r c . A Monochromatic I l l u m i n a t o r o f Adam F i l g e r , Ltd.., fitted  w i t h q a a r t z p r i s m and l e n s e s and r e a d i n g d i r e c t l y t o  wave l e n g t h s was used t o r e s o l v e t h e l i g h t  t  f h e instrument  was s t a n d a r d i s e d fey r e a d i n g s on t h e s o d i n a 33 l i n e . Par&aoeeia fitted  were p l a c e d i n a q u a r t z c e l l which  I n t o the s l o t o f t h e l o n o c h r o a a t l c i l l u m i n a t o r , t h e  c o n t r o l b e i n g kept i n a s i m i l a r c e l l i n t h e d a r k . f h e e f f e c t of v a r i o u s monochromatic l i n e s o f t h e l l e r e a r y A r c Spectrum on t h e r a t e o f d i v i s i o n o f t h e organism was a s c e r t a i n e d and a l s o t h e e f f e c t on t h e r a t e o f p l a s m o l y s i s .  PIBCOBSIQH OP FPBULTS. 1.  Rate o f growth.  { fable  1.)  from t h e accompanying g r a p h f P l a t e 1.)  i t will  PMfB I  Stimulation  and R e t a r d a t i o n o f Growth i n Paranaoeeirea expressed as  namher o f Paramoeeia g r e a t e r o r l e s s than i n t h e c o n t r o l .  9. fABLB I B f f e e l o f various wave- lengths of • l i R h t on t h e r a t e of d i t r i l ion ox J?araaoeeinffl oaadatna. lave I•gngth in &. •  Somber o f Para»eeaina a.iter, © boars.  I r r a a iated  Control  Biff.  Hamber of Paraaoeeinm a f t e r 24 h o u r s . Irradiated  Control  Biff  6152  ' 13.4  18*8  0.6  15. &  14.35  0.85  68195769  12.85  12.85  0.0  17.0  15.4  1.6  49684916  9.1  13.0  -3.9  15.0  16»8  —1.0  43594348  14.1  13.0  1.1  19.1  17.0  2.1  40783984  13.2  13.4  -0.8  12.9  14.7  -1.8  3881  11. 4  11.3  0.1  12.7  13*3  -0.6  36633666  18.1  11* 3  0.8  13.6  12.4  1.2  33523348  11 • §  10.5  1.0  12.9  12.2  0.?  3138  12* 3  12 .1  0.2  13.4  12. 9  0.5  3088  12.0  13.0  -1.0  14.2  15.5  -1.3  8967  10.4  11*6  -1.8  10.3  12* 2  -l.f  8804  9.9  11» 9  —2 .0  9.6  13*5  -3.9  8700  10.4  12 . &  -1.8  9*6  14.3  -4.7  £535  10.0  14*8  -4.2  7.8  14.8  -7.0  2054  9.9  13.8  —3*9  8.6  14.0  -5.5  10. fee seen that there are two regions which d e f i n i t e l y and c o n s i s t e n t l y i n h i b i t the growth r a t e of Paramoeeina; that region areand 4960 A , the indigo bine end o f the s p e c t r n a , o 0  and  that beyond 3132 A , the u l t r a v i o l e t and f a r u l t r a  violet portion,  there a l s o seem to fee three regions o f  s t i m u l a t i o n , the y e l l o w orange, 5916 A  and the v i o l e t i n -  0  d i g o , 4359 A , and a s l i g h t s t i m u l a t o r y e f f e c t a t t h e be0  g i n n i n g o f the u l t r a v i o l e t 3821 A°-3348 A , feat at no time 0  does the magnitude o f s t i m u l a t i o n approach that o f the most extreme r e s u l t s o f r e t a r d a t i o n . l a v e length a i d not i n t e n s i t y i s the determining  factor,  f o r the greatest r e s u l t s a r e obtained w i t h many o f the l i n e s of l e a s t i n t e n s i t y ( ex. 2054 A°, 4960 A ) .  In cases o f  0  s l i g h t and t r a n s i t o r y  i n h i b i t i o n of growth r a t e , a period  ©f a c c e l e r a t i o n f o l l o w s ( 5819 A , 3821 A 0  0  and 3132 A ° ) ,  s u f f i c i e n t to r e s u l t i n an a c t u a l increase i n the number of Irradiated  i n d i v i d u a l s as compared t o non-irrad iated  control;  shoald the period of i n h i b i t i o n be prolonged the organism does not recover i t s normal growth, r a t e and i n extreme cases t o t a l inhibition results  { 3022 A  0  onwards J .  these f i n d i n g s correspond i n general w i t h the r e s u l t s of Bovie and Hughes (16) f o r r a y 2800 A°. where c y t o l y s i s o f Paramoeciua was caused by prolonged i r r a d i a t i o n and exposure Insufficient  t o produce c y t o l y s i s caused i n h i b i t i o n o f c e l l  d i v i s i o n followed fey s t i m u l a t i o n .  Ho e f f e c t w i t h s l i g h t  11. i r r a d i a t i o n s a y be dne penetrate  and  t o the i n a b i l i t y o f the r a y s  e f f e e t the  2.  to  mieleus.  I f f e a t on r a t e o f Plasmo l y s i s . ( f a b l e  f h e e f f e c t s of monochromatic l i g h t on the of plasotolysis i n Paraaoeciua, using varying of BaCl.show t h a t t h e r e a r e a g a i n two regions  of greatest  effect.  I n no case was  8.) rate  concentrations  o r perhaps t h r e e  ( P l a t e s £ and  3.)  a d e c r e a s e i n r a t e n o t i c e d , even  w i t h t h e l i n e s c a u s i n g most e f f e c t e i t h e r i n s t i m u l a t i o n o r i n h i b i t i o n o f growth;  a l l wave l e n g t h s s e e m i n g l y h a v i n g some  s t i m u l a t o r y e f f e c t on p l a s m o l y t i c r a t e .  A f t e r s i x hours i r -  r a d i a t i o n t h e r e s u l t s a r e a l i t t l e e r r a t i c , t h o s e l i n e s around 365GA -3Q£2 A° b e i n g c o n t r a d i c t o r y , but even a f t e r t h i s e  short  t i m e t h r e e r e g i o n s ©anaing i n c r e a s e i n r a t e can be n o t e d . A f t e r t w e n t y - f o u r h o a r s t h e r e g i o n s 4960 A , 0  4078 A , 0  and  3022 A° down to t h e f a r u l t r a v i o l e t cause the g r e a t e s t i n crease i n r a t e . These r e s u l t s when compared w i t h the e f f e c t on the r a t e o f growth o f Paramoeelurn show a marked ( see p l a t e s 2 and  3.)  the r e g i o n s c a u s i n g most I n h i b i t i o n i n  growth c o r r e s p o n d i n g t o t h o s e c a u s i n g r a t e of p l a s m o l y s i s .  similarity,  greatest  Those r e g i o n s c a u s i n g  increase i n  little  effeet  on g r o w t h , or s t i m u l a t i n g s l i g h t l y , correspond w i t h t h o s e c a u s i n g the l e a s t i n c r e a s e i n the r a t e o f  plasmolysis.  12  TAMIM 2/  E f f e c t o f v a r i o a s wave-lengths of l i g h t on t h e r a t e o f p l a s a o l v s i s o f garastoeclnja eamdatoa* Increase i n r a t a of increase" over c o n t r o l rate.' lav  a expressed as percentage  Soilnffl C h l o r i d e S o l u t i o n s 0.7 Molar 0.3 Molar  th  6 hoars  6 hours  24  6152  30  29  §8195769  19  21  49604916  41  39  33  48  4359 4348  80  34  26  33  40783984  39  39  32  46  3881  26  40  35  45  36633656  88  27  40  25  33583342  9  11  34  31  37  20  hours  37 26  24 hours 41 27  3132  86  3022  44  18  35  24  296?  34  36  37  39  2804  40  51  34  43  2054  51  51  58  57  81  Comparison o f the e f f e c t oa growth o f P a r a a o e e i n a ano  on p l a s m o i y s l s In sod iota c h l o r i d e solutions.  P l a s r a o l y s i s expressed as percent i n c r e a s e i n r a t e  Comparison  of the effeet  and on p l a s m o l y s i s  on growth of Pftramoeciam i n sodlnm  chloride  eolations.  P l a s a o l y s i s expressed as p e r c e n t i n c r e a s e  i n rate  13  f h a s i t would  seem t h a t t h e r e i e a c o r r e l a t i o n  Between d i v i s i o n r a t e and p l a s m o l y t I c r a t e I n Pararaoec iua. Packard  (E6j w o r k i n g on t h e same organism  t h a t t h e p e r m e a b i l i t y o f t h e c e l l s exposed  t o l i g h t was  g r e a t e r than t h e c o n t r o l s kept i n d a r k n e s s , p e r m e a b i l i t y was a l s o demonstrated  found  f h e change i n  i n c e l l s exposed t o  monochromatic red l i f c h t , becoming g r e a t e r as t h e wave l e n g t h s h o r t e n e d and r e a c h i n g a maximum i n t h e near  ultra-violet.  A change i n t h e r a t e o f p l a s a a o l y s i s may be caused by v a r i o u s f a c t o r s .  Blacknan and  P a i n e (2) r e c o r d  th«t l i g h t has a marked e f f e c t on t h e p e r m e a b i l i t y o f t h e c e l l s o f Mimosa pud i c a t o e l e c t r o l y t e s , p r o b a b l y due t o t h e " d i s a p p e a r a n c e o r i n a c t i v a t i o n o f t h e osmotic s u b s t a n c e s o f the c e l l " .  On the o t h e r hand l i g h t b e i n g absorbed  by the  c e l l membrane, i t i s p r o b a b l e t h a t t h e membrane I t s e l f changes I t s p e r m e a b i l i t y , perhaps due t o t h e a c t i o n o f l i g h t on t h e l i p o i d s u b s t a n c e s w h i c h a r e b e l i e v e d t o be one o f i t s constituents. I t c a n be d e f i n i t e l y s t a t e d , however, t h a t c o n d i t i o n s accompanying moderate i n c r e a s e i n r a t e of p l a s m o l y s i s a l s o accompany i n c r e a s e i n r a t e o f c e l l d i v i s i o n and t h a t condition© accompanying extreme i n c r e a s e i n r a t e o f p l a s m o l y s i s r e s u l t i n decrease I n growth.  14.  Experimental Berk. 2.  f h e e f f e e t o f B a f l i a n t Energy oa the growth  and s.poralatlon o f G o l l e t o t r i c h a m ghomoides. fhe  experiment8 to he d e s c r i b e d d e a l w i t h t h e  e f f e e t o f r a d i a n t energy on 1) growth 2) s p o r n l a t i o n , o f G o l l e t o t r i c h t t m p b o o o i d e s , a common cause o f r i p e r o t o f tomatoest19). P o t a t o d e x t r o s e a g a r was used a s t h e n u t r i e n t medium, s i n g l e p l a n t i n g s b e i n g made on t h e agar and a l l o w e d to grow f o r t h r e e days b e f o r e i r r a d i a t i o n . g x p e r l a e n t 1. I r r a d i a t i o n w i t h f a l l l i g h t from a Mercury arc lamp. 1.  S i n g l e s p o r e c u l t u r e s o f t h r e e days growth  were exposed t o t h e f u l l l i g h t , from a q u a r t s mercury lamp at  a d i s t a n c e o f 12 cms. from the s o u r c e , p a r t o f the c u l t u r e  b e i n g p r o t e c t e d by a h a l f c i r c l e o f b l a c k c a r d b o a r d . The l i n e o f s e p a r a t i o n was e v i d e n t w i t h i n a few minutes o f i r r a d i a t i o n , caused by t h e c o l l a p s e o f the a e r i a l mycelium i n t h e exposed p a r t ; i n a l l c a s e s growth was r e t a r d e d and s p o r u l a t i o n accelerated i n the area i r r a d i a t e d . ( P l a t e 4 ) . 2.  S i n g l e s p o r e c u l t u r e s o f t h r e e days growth  were i r r a d i a t e d f o r p e r i o d s o f t h r e e minutes t o t h i r t y seconds w i t h t h e l i g h t from a q u a r t s mercury lamp. I t was found t h a t I r r a d i a t i o n f o r t h r e e minutes caused s t u n t i n g  flATM 4 »  I f f e e t on a p o r u l a t l o n ana growth o f I r r a d i a t l n g " h a i f o f three-aay c u l t u r e s w i t h t h e a e r e t i r y vapour a r c s p e c t r u m , lower s i d e exposed. npper s i d e n o r m a l .  A.  C u l t u r e , 14 days a f t e r 4 rain, e x p o s u r e .  3.  Culture,  3 days a f t e r 15 m i a . exposure.  15. from w h i c h the c o l t ore d i d not a f t e r w a r d s r e g a i n the normal growth r a t e .  ( f a b l e 3.)  I r r a d i a t i o n o f from f i f t e e n  seconds  to  two minutes caused s t u n t i n g i n d i r e c t r e l a t i o n to the t i m e  of  e x p o s u r e , but f r o m w h i c h t h e c u l t u r e r e c o v e r e d and  a t e l y exceeded  the c o n t r o l i n growth*  ultim-  D i r e c t i r r a d i a t i o n of  the  t h r e e day c u l t u r e r e s u l t s a f t e r about t w e n t y - f o u r hours i n  the  f o r m a t i o n o f numerous a c e r v u l i , m o s t l y i n clumps, t h e max-  imum development to  of acervuli taking place after t h i r t y  one minute e x p o s u r e , l o n g e r p e r i o d s o f i r r a d i a t i o n  e r a t i n g development  but i n s m a l l e r q u a n t i t i e s .  seconds accel-  Development of  a c e r v u l i i n t h e c o n t r o l d i d not. u s u a l l y t a k e p l a c e u n t i l t h e e i g t h da# o f growth. 3.  I r r a d i a t i o n o f a s u s p e n s i o n of s p o r e s .  A s u s p e n s i o n o f s p o r e s i n s t e r i l e w a t e r was full  i r r a d i a t e d w i t h the  i l l u m i n a t i o n from a q u a r t s mercury lamp and t h e s p o r e s  t h e n p l a t e d out on b a c t o p o t a t o d e x t r o s e a g a r . I r r a d i a t i o n o f s p o r e s f o r p e r i o d s of more t h a n t h i r t y seconds caused an i n t e n s i v e i n i t i a l s t u n t i n g from which the  c u l t u r e d id not r e g a i n normal growth r a t e .  Irradiation for  f i f t e e n seconds a l s o caused i n i t i a l s t r m t i n g but the r a t e g r a d u a l l y approached norma}..  I t would  growth  seem from t h e s e  r e s u l t s t h a t t h e e f f e c t of i r r a d i a t i o n i s more d r a s t i c i n the c a s e o f the s p o r e s t h a n the growing rayeeliwffi, but i t should be born i n mind t h a t many of t h e s p o r e s i r r a d i a t e d may  have  g e r m i n a t e d , and t h a t t h e g e r m i n a t i o n t u b e was exposed t o t h e r a y s w h i c h proved more h a r m f u l t o i t t h a n t o the o l d e r mycelium  16  msm  a.  B f f e e t o f f a l l l l l a m i n a t i o n w i t h the mercer? a r c lamp npon  Exposure  Control 3 minutes 2 minutes 1 alnnt© 30 seconds 15 seconds  Bays 4th#  S i?,e o f c o l t o r e I n  5th.  6th  7th.  3.23 2.5 2.15 2.4 . 2.1 2.9 2.25 3.0 P. 3 . 3.1  4.1  4.3 4.1 . 4.3 4.7 4*9 4.8  * « »* *  «  a.4  2.1  TA3LE  -  —  _ V  .  ems *  8th.  9th.  1 0 t h . 11th.  5.6  6.3 5.6  7.1 6.6 7.1 7.3 7.5 7.4  5.1 5.35 6.6  5.7 5.7  6.1  6.3 6.5 6.5  7.8 7.2 8.2 7.9 8.3 7.9  a  4.  E f f e c t o f f a i l I l l u m i n a t i o n w i t h t h e mercury a r e lamp anon C o I I e t o t r t c h q g i B P ore' "s^epani'lona. Expos o r e  S l a e o f c u l t u r e I n cms. Bays ¥4h.  Control 2 a i n n t ee 1 minute 30 seconds 16 seconds  £.4 2.0 1.95 2.1 2.1  5th. 3.28 2.7 2.7 2.9 3* X  6th.  -*  4.1  -  7th.  8th.  9 t h . 10th  4.8 ft.O 4.4 4.6 4.7  5.6 5.1 5.1 5.2 5.5  6.3 5.9 5.9 6.1 6.2  7.1 6.5 6.6 6.7 7.0  11th 7.8 7.2 7.4 7.5 7.8  17  of t h e t h r e e day c u l t u r e s . I n a l l cases development o f a c e r v n l i was h a s t e n e d , b e i n g noted i n t h e i r r a d i a t e d c u l t u r e s e v e r a l days b e f o r e t h e i r development i n t h e c o n t r o l ,  iiaximum development  seemed to t a k e p l a c e i n c u l t u r e s i r r a d i a t e d f o r not more than t h i r t y seconds.  Experiment £» I r r a d i a t i o n o f s p o r e s I n s u s p e n s i o n by fflonoo.hr oaat l e l l ^ h t . A s u s p e n s i o n o f s p o r e s i n s t e r i l e w a t e r was placet i n a quarts container  i n the s l i t  of t h e Mono-  c h r o m a t i c I l l u m i n a t o r and was i r r a d i a t e d w i t h v a r i o u s monoc h r o m a t i c l i n e s of t h e mercury a r c s p e c t r u m . The e f f e c t of i r r a d i a t i o n o f t h e s p o r e s by s p e c i f i c wave l e n g t h s was s e e m i n g l y c o n f u s i n g u n t i l into four classes. C l a s s 1.  divided  ( f a b l e 5.J  3ti.anls.tion continuous. 6819 A ° , 4078 A ° , 3656 A° and 3348 A° cause  s t i m u l a t i o n of growth c o n t i n u i n g p e r i o d , except i n the c a s e o f  throughout the e l e v e n day  3342  A  0  where t h e maximum  s t i m u l a t i o n i s reached a t t h e f o u r t h d a y . G l a s s £.  Retardation  followed  by s t i m u l a t i o n .  I r r a d i a t i o n by l i n e s i n t h i s c l a s s r e s u l t s i n a p r i m a r y r e t a r d a t i o n o f s m a l l degree f o l l o w e d  by s t i m -  u l a t i o n w h i c h i s g e n e r a l l y i n inverse? p r o p o r t i o n i n i t i a l retardation.  t o the  T h i s e f f e c t i s caused by wave l e n g t h s  18 fABX.S 5. S p e o i f i o e f f e c t s o f Renoohreaatio l i g h t apon the growth o f C o l l e t 6 trlafaqg, s p o r e s u s p e n s i o n s * l a v e length, Jfccp. S i s e m A ** " In hrs." 4th. ©oat r o i 2.4 C l a s s 1. Wave l e n g t h s 1 2.4 £ 2.6 A  i  o f c o l t ore i n cms. " ' 5 t h . 6 t h . 7 t h . 8 t h . 9th« 3.28 4.1 4.8 5.6 6.3 7.1 causing consistent stimulation 3.2 4.0 5*0 5.7 6.3 7.2 3.5 4.5 5.7 6.4 7.0 7.4  7.8 8.0 8.1  40783984  1 2  8.6 2. 6  3.2 3.1  4.0 4.1  4.9 5.2  5.6 6.6  6.9 7.6  7.5 8.2  8.1  36563660  1 2  3.0 £.8  3.6 3.5  4.3 4.2  4.7 5.3  5.6 6*0  6.3 7.3  7.1 7.8  7.8 8.4  11  2*9  3.4  3.9  4.8  5,8  6.3  7.1  7.8  3342  C l a s s 2. 'aye ^ 2 6  leg~  3881  l e n g t h s causing, r e t a r d a t i o n f o l l o w e d STB 3.1 4.1 4.8 a , 6'673" 2.3 3.0 4.1 5.0 5.8 7.2  av s t i m u l a t i o n , 7.1 "~ ' 8.0 8.5  1 2  2.3 2* 3  3.1 3.1  4.5 4.2  4.9 5.0  5.4 6.0  5.8 6.8  7.0 7.6  7.8 8.5  31323126  I 2  2.1 2.3  2*8 3.2  4.0 3.7  4.9 4.8  5.9 6.1  6.7 6.8  7.6 7.6  8*1 8.6  2967  1 2  2*2 1.9  3.1 2.8  3.9 4.0  4.8 5.0  5.7 5.5  6.7 6.6  7.4 7.4  8.1 8.4  2700  1 8  3.2 3.2  4.0 4.0  4.7 4.7  6.1 6.1  6*4 6.5  7.2 7.3  8.3 8.5  C l a s s 3*  Wave l e n g t h s c a u s i n g complete r e t a r d a t i o n .  49604916  1 2  1.9 1.9  8*9 2*7  3.8 3.6  4.8 4.8  5.6 5.5  6.3 6.1  7.0 6.9  7.7 7.6  3028  1 2  1.5 1.4  2.1 2. 1  3.6 2.8  4*2 3.5  5.2 4.2  5.5 5.7  6*0 6.3  7.0 7.2  2804  1 £  1.6 1* £  2.2 £.1  2.8 2.6  3*3 3.4  4.4 4.2  5.4 4.7  6.1 §.7  7.7 6.5  2064  1 2  1.5 1*5  2.5 2.8  3*4 8.8  4.0 3.5  5.0 4.6  5.9 5.1  7.0 6.0  7.7  1849  1 £  2.0 1.8  3.1 2.5  3.8 3.3  4.4 4.8  5*5 5.1  6.1 6.1  7.1 7.0  7.8 7.7  -  19. 6152 A , 0  3JL32 A , 0  G l a s s 3.  3821 A ° , 2700 A  0  and 8967 A ° .  Continued r e t a r d a t i o n . Continued r e t a r d a t i o n i s caused by w a v e - l e n f l t b s : -  4960 A , 0  3028 A ° , 8804 A , ft  o f the mycelium i s stunted  2054 A  0  and 1849 A ° .  fhe growth  d u r i n g the f i r s t few days  after  I r r a d i a t i o n , and t h e normal growth r a t e i s n e v e r reached t h e p e r i o d measuredthe  over  I t w i l l be n o t e d , however, t h a t where  i n i t i a l s t u n t i n g i s not so i n t e n s e the growth r a t e ap-  p r o x i a a t e s more c l o s e l y t o n o r n a i (2804 A C l a s s 4.  go  0  and  1849  A }. 0  effect. 4359 A  appreciabie  does no a f f e c t the r a t e o f growth t o any  eztent.  ( See P i a t e 5.).  DISCUSSIQW OF  R^DXffS.  f h e e f f e c t of d i r e c t i r r a d i a t i o n of g r o w i n g c u l t u r e s and  of s u s p e n s i o n o f s p o r e s In s t e r i l e w a t e r , on the  • r a t e o f growth o f the o r p a n l o n can i n both eases be seen t o be p r o f o u n d l y a f f e c t e d .  I t i s apparent t h a t the i r r a d i a t i o n  o f s p o r e s has a a c r e narked e f f e e t t h a n the i r r a d i a t i o n o f t h e t h r e e day c u l t u r e s , e m p h a s i s i n g the f a c t t h a t t h e e f f e c t must be p u r e l y p r o t o p l a s m i c , nnd not due t o changes i n t h e c u l t u r e ned i n n causes by  irradiation.  I t w i l l be seen fro© P l a t e 6 t h a t when r e t a r d a t i o n of growth extends e v e r t h e t e n day p e r i o d , s p o r u l a t i o n  B l A f B S.  f h e p r o g r e s s i v e e f f e c t o f v a r i o u s linmm o f the mercury are speetrnm on t h e growth o f OoTle'r"-t o t r l c h o s i phomoides a t t i n e i n t e r v a l s n-p "WZZ* and t e n d a y s . ' • —= =•  B f f e e t expressed as cms. i n c r e a s e or decrease over c o n t r o l c u l t u r e growth.  PXAgB 6  The r e l a t i o n s between t h e e f f e c t of monoohronat 1c i l f l h t on the g r o w t h and s p o r n l a t l o n o f C o l l e t o t r i e h n s i . t h e c o n t r o l r e p r e s e n t e d as zero.  80 i s hastened.  I t may  he n o t e d , however, t h a t when the  mercury a r c spectrum Is used f o r i r r a d i a t i o n ,  full  sporulation  i n c r e a s e * w i t h exposure's o f from f i f t e e n seconds t o two  min  u t e s i n d u r a t i o n , hut d e c r e a s e d when exposures were f o r longer time  than two  minutes,  of s p o r u l a t i o n  f h l s seems to i n d i c a t e t h a t  i s an  Inverse expression  o f growth  r a t e o n l y w i t h i n c e r t a i n l i m i t s and  that there  timum amount o f I r r a d i a t i o n c a u s i n g  sporulation.  i s an  I t w i l l a l s o he noted f r o m P l a t e 6 t h a t t a r d a t i o n and  the  op-  re-  extreme s t i m u l a t i o n o f growth a r e accompanied  by e a r l y s p o r u l a t i o n , w h i l e  i r r a d i a t i o n causing  Inter-  mediate s t i m u l a t i o n o f growth has no a p p r e c i a b l e  effect  on the time o f development of a c e r v u l i . S t e v e n s (34) w o r k i n g on v a r i o u s f u n g i found t h a t i r r a d i a t i o n "almost i n s t a n t l y i n i t i a t e s the opment of r e p r o d u c t i v e  stnuctures  devel-  i n g r e a t numbers where  t h e y would not have o c c u r r e d w i t h o u t i r r a d i a t i o n " . g e s t s t h a t t h e s e r e s u l t s may of g r o w t h caused by the r a y s . and  B a i l e y (30)  be due  He  sug-  t o a sudden I n h i b i t i o n  On t h e o t h e r hand Ramsey  I r r a d i a t e d M a e r o s p o r l n a and  Pnsariura  and  s t a t e t h a t t h e r e i s a d e f i n i t e s t i m u l a t i o n of s p o r u l a t i o n caused by u l t r a - v i o l e t l i g h t , w h i c h s e e i s to be a d i r e c t r e s u l t of s t i m u l a t i o n r a t h e r than the  indirect r e s u l t of o  inhibition.  obtained  2535 A . 0  G r e a t e s t s p o r u l a t i o n was  S t i m u l a t i o n o f spore p r o d u c t i o n  8535 A° but t h e r e was  at 8800 A -  o c c u r r e d below  some l e t h a l e f f e c t a l s o and  re-  21. t a r d a t i o n o f m y c e l i a l development,  f h e s e a u t h o r s a l s o found  t h a t l o n g exposure t o d i r e c t s u n l i g h t through f i l t e r s m i t t i n g no l o w e r t h a n 3132 both i n , iCacrosporium  and  F u l t o n and  A°  induced abundant  trans-  sporulatlon  Fusarium. Coblent?, (8)  i r r a d i a t e d the  spores  o f eiany s p e c i e s of f u n g i w i t h u l t r a v i o l e t l i g h t and that the l e t h a l e f f e c t r e s u l t e d a f t e r comparatively  found short  exposures, although the time required t o V i l l varied  with  the t y p e o f f u n d u s . D i l l o n Weston (6) a l s o r e p o r t s t h a t  thw  p r i m a r y r e a c t i o n on F u s a r i u m i s t h a t of r e t a r d a t i o n , subsequently,  however, i r r a d i a t e d c u l t u r e s s p o r u l a t e d  a b u n d a n t l y , whereas the n o n - i r r a d i a t e d c u l t uses  very  sporulated  sparsely. From t h e s e v a r i o u s e x p e r i m e n t s i t #111  be  seen t h a t l i g h t has a profound e f f e c t on the p r o d u c t i o n spores,  of  but whether t h i s a c c e l e r a t i o n of s p o r e p r o d u c t i o n  is  t h e d i r e c t r e s u l t o f i r r a d i a t i o n or the r e s u l t o f r e t a r d a t i o n o f the m y c e l i a l g r o w t h c a u s i n g a s t i m u l a t i o n of s p o r e p r o d u c t i o n has not  yet been d e t e r m i n e d .  U p a p a r l s o n of the e f f e c t s o f Monochromatic L i g h t on a n i m a l protoplasts ( Pararaoecina) and  on p l a n t p r e t o p l a s a ( G o l l e t o t r l o f a i p s )  In o r d e r t o f a c i l i t a t e comparison we w i l l d e a l w i t h t h e r e s u l t s i n e l a s s e s , i . e . , C l a s s 1, t h o s e l i n e s c a u s ing  s t i m u l a t i o n i n C o l l e t o t r i e h n m ; C l a s s 2, those l i n e s  that give r i s e t o r e t a r d a t i o n followed C o l l e t otrichnm  by s t i m u l a t i o n i n  and C l a s s 3, l i n e s t h a t cause constant  retard-  ation i n Colletotriohu i . { See P l a t e 7 ) . C l a s s 1.  5819 A ° , 3656 A ° , 3342 A  continued  s t i m u l a t i o n i n C o l l e t otrichnm.  and  0  and 4078 A  0  show  Of t h e s e 5819 A  0  3656 A° show s i m i l a r s t i m u l a t i o n i n Paramoeoiuta, 4078 A  g i v i n g s t i m u l a t i o n followed C l a s s 2.  by r e t a r d a t i o n .  6158 A ° , 3132 A ° , 3821 A  to r e t a r d a t i o n followed  0  0  and 8967 A  0  give  rise  by s t i m u l a t i o n i n C o l l e t o t r i c h u m .  In  Paramo©oiurn t h e e f f e c t s o f t h e f i r s t t h r e e l i n e s a r e n o t t h a t o f r e t a r d a t i o n but i t w i l l be n o t i c e d t h a t a f t e r twentyf o u r hours t h e growth i s more t h a n t h a t r e p o r t e d  f o r the  c o n t r o l , showing t h a t t h e e f f e c t i s one o f l e s s to more s t i m u l a t i o n . 2967A0 C l a s s 3.  t  however, caused r e t a r d a t i o n i n Paramoeciurn.  4960 A , 3022 A , 2804 A , 2535 A° and 2054 A 0  0  0  0  a r e found t o r e t a r d , growth i n both Paramoecium and C o l l e t otrichum.  S i m i l a r r e s u l t 8 were r e p o r t e d  by H u t c h i n s o n and  l e w t o n (20) i n t h e i r work on Y e a s t . I t can be seen from t h e s e r e s u l t s t h a t i s a decided ( P l a t e 7}  there  c o r r e l a t i o n between t h e two s e t s o f r e s u l t s . I n C l a s s 3, where i n h i b i t i o n i s r e p o r t e d  in a l l  c a s e s i t would seem t h a t the f a r u l t r a v i o l e t has a p r o found e f f e c t on b o t h a n i m a l and p l a n t p r o t o p l a s m and t h a t o o c e r t a i n r e g i o n s i n t h e v i s i b l e ppectrum, 4960 A -4916 A ,  P M E  7.  Diagram o f the o o a n a r a t l T a e f f e c t s o f monochromatic l i g h t on C o l l e t e t r i c h n m ana. Paraaoeclna, the c o n t r o l representee" a s a e r o .  r CO  esq  to  S3 X*X-X»X  i to  [,-XXXXX,  tn <  "US !/>/yyy/  -Q  to  3 1  be  a: CO  2 CJ Lu  a LU CD  H •<  —CJ  ii<  i —  Jm  23 a l s o e x e r t an W e n t l e a l e f f e e t . Where the e f f e c t s a r e not. so f a r - r e a c h i n g , d i f f e r e n c e s may he found Between t h e two r e s u l t s , p r o b a b l y due t o t h e v a r i e d s u s c e p t i b i l i t i e s o f t h e organisms used and t h e d i f f e r e n c e i n amount o f l i g h t absorbed by the p r o t o p l a s m .  24  SOMtARY Of R*:SgLTS. 1.  There a r e two r e g i o n e i n t h e M e r c u r y A r e Spectrum  that  d e f i n i t e l y i n h i b i t the growth r a t e of Paramoecina caudatnm:~ 4960 A , 3132 A 0  2.  0  and a l l s h o r t e r wave l e n g t h s  t h e r e a r e two r e g i o n s  used*  o f the d e r o u r y A r c Spectrum  that  cause s t i m u l a t i o n i n Paramoecium c a u d a t u a growth r a t e : 5819 A  0  and 4359 A . 0  3.  Under the c o n d i t i o n s  not  i n t e n s i t y i s the d e t e r m i n i n g f a c t o r .  4.  S l i g h t t r a n s i t o r y i n h i b i t i o n i n growth r a t e i s u s u a l l y  followed  B?- a p e r i o d  o f t h e s e e x p e r i m e n t s wave l e n g t h and  o f a c c e l e r a t i o n both In Paramoecium and  Colletotriohma. 5.  A l l l i n e s of t h e Mercury A r c Spectrum t& ed i n t h e s e  e x p e r i m e n t s caused an I n c r e a s e i n t h e r a t e o f p l a s m o l y s i s of Paramoecium can.datua i n Sodium C h l o r i d e  solutions.  6.  fhere  and  p asmolytic  7.  I r r a d i a t i o n o f three day c u l t u r e s o f C o l l e t o t r i c h u r n  1  i s a d e f i n i t e c o r r e l a t i o n between d i v i s i o n r a t e r a t e i n Paramoecium caudatum.  phomoides caused r e t a r d a t i o n o f growth and a c c e l e r a t i o n o f s p o r u l a t i o n when t h e f u l l  i l l u m i n a t i o n from a Quarts  M e r c u r y A r c lamp was used. 8.  S u s p e n s i o n s of s p o r e s of O o l i e t o t K i e h n m phomoid es  i r r a d i a t e d w i t h t h e M e r c u r y A r e Spectrum haa growth r a t e retarded  and s p o r u l a t i o n  acceleration.  £5  9.  fhe e f f e c t of monochromatic l i g h t on s p o r e s u s p e n s i o n s  i s of t h r e e k i n d s , followed 10.  i . e . . s t i m u l a t i o n o f growth, r e t a r d a t i o n  by s t i m u l a t i o n and  Retardation  r e t a r d a t i o n o f growth.  or extreme s t i m u l a t i o n of g r c r t h i s  accompanied by e a r l y s p o r u l a t i o n , i r r a d i a t i o n c a u s i n g s l i g h t s t i m u l a t i o n o f g r o w t h h a v i n g no a p p a r e n t e f f e c t  on  development of a c e r v u l i . 11.  A c o r r e l a t i o n can  on P a r a m o e c i u i and  be seen between t h e e f f e c t of  on C o l l e t o t r i o h u m ,  but  light  i t i s Impossible  to say a t t h i s stage whether t h i s holds f o r a l l animal p r o t o p l a s m as compared w i t h p l a n t  protoplasm.  £6. PAR? B. fm  OF RAPlAKf MBRQY OS M.^YMR ACf 101  ISTBODPCflOH I t would of t h i s work and  seem from r e s u l t s obtained  i n e x p e r i m e n t s undertaken by  a u t h o r s , t h a t l i g h t has a profound  In P a r t A other  e f f e c t on b o t h p l a n t •  mi  a n i m a l p r o t o p l a s m and t h a t t h i s e f f e c t must be  on ©ne o r more f a c t o r s . to  be d e t e r m i n e d .  \  based  What t h e s e f a c t o r s a r e has y e t  S e v e r a l t h e o r i e s , however, have been  propounded. According  t o F. I». S a t e s (9) t h e e f f e c t o f  measured monochromatic u l t r a v i o l e t energy used t o k i l l b a c t e r i a shows c h a r a c t e r i s t i c and s i m i l a r c u r v e s a t each wave l e n g t h , but an a p p r e c i a b l e amount o f energy must be i n c i d e n t on the b a c t e r i a b e f o r e any o f them succumb, and w i d e l y d i f f e r e n t i n t e r m i t i e a of energy a r e r e q u i r e d t o produce t h e s e c u r v e s a t d i f f e r e n t wave l e n g t h s ,  fhe re-  c i p r o c a l s of these curves are s i m i l a r to the a b s o r p t i o n curves o f c e r t a i n d e r i v a t i v e s of the n u c l e o - p r o t e i n s a r e r e l a t e d t o c e l l growth and  reproduction.  which  T h i s con-  c l u s i o n i s supported by t h e known f a c t t h a t t h e a c t i v e agent o f c h i c k e n t umours i s a s s o c i a t e d w i t h a n n c l e o - p r o t e i n , w h i c h , i n a l l c a s e s of sarcomatous Feulgen r e a c t i o n .  c e l l s , gives a p o s i t i v e  I t i s t h e r e f o r e p o s s i b l e t h a t the  e f f e c t and the i n h i b i t i o n o f c e l l d i v i s i o n caused  lethal  by u l t r a  2?  v i o l e t l i g h t may he r e l a t e d t o I t s a c t i o n on t h e n u c l e o proteins of the nucleus. I. I . H a r r i s and R. S. E o y t e {13} prepared a pure c u l t u r e of Pararaoecinm and a second i n a s o l u t i o n o f a r o m a t i c amino a c i d s ,  culture  suspended  f h e suspension  was i r r a d i a t e d d i r e c t l y and t h e pure c u l t u r e through a s h i e l d formed  by a l a y e r o f t h e a c i d s o l u t i o n and i n both  c a s e s t h e t o x i c i t y was d e c r e a s e d .  T h i s r e s u l t suggested  the p o s s i b i l i t y t h a t the s u s c e p t i b i l i t y o f p r o t o p l a s m t o ultra-violet light  i s conditioned  by t h e s e l e c t i v e ab-  s o r p t i o n o f t h e t o x i c r a y s by t h e a r o m a t i c amino a c i d r a d i c a l s o f the p r o t e i n s . H u t c h i n s o n and A s h t o n (17} have suggested  that  resonance phenomena may be i n v o l v e d , and t h a t c e r t a i n p a r t i c l e s i n a c o l l o i d a l o r o t h e r s t a t e possess v i b r a t i o n p e r i o d s such t h a t t h e i r movements a r e a f f e c t e d by the s p e c i f i c f r e q u e n c e s o f t h e r a d i a n t energy* subsequent  On t h i s  basis  e f f e c t s may d i f f e r from t h e i n i t i a l ones owing  to associated  changes i n t h e c o l l o i d a l c o n d i t i o n .  As t h e r e i s a c e r t a i n d ependance o f ensyme a c t i v i t y on t h e c o l l o i d a l s t a t e i t was thought t h a t t h e e f f e c t o f r a d i a n t energy on enzyme a c t i o n might throw some l i g h t on t h e s e b a s i c f a c t o r s c a u s i n g l e t h a l or s t i m u l a t o r y e f f e c t s on p r o t o p l a s m .  28. BETiBw OF Lxyrmggas.  Light  i n f l u e n c e s enzyme a c t i o n s by  t h e ensyme i t s e l f or fey m o d i f y i n g f h i e s and d iffused  i t s activity.  Wiekern (82) found t h a t the l i g h t on p e r o x i d a s e and  inhibitory  blood  f o l l o w i n g o r d e r , white>blne> red > d a r t .  destroying  Lockman, e f f e c t of  c a t a l a s e was  i n the  H e i n l e (31)  fonnd  t h a t t h e methylene bine formaldehyde r e d u c t a s e r e a c t i o n of cow's m i l k was G e t c h e l l and  reduced by the a c t i o n o f u l t r a v i o l e t  W a l t e r (10) s t a t e d t h a t u l t r a v i o l e t  light.  light  more p o t e n t i n the d e s t r u c t i o n o f p e r o x i d a s e a c t i v i t y e i t h e r X - r a y s , radium o r v i s i b l e P i n c u a s e n (87) l i g h t on d i a s t a s e and  reaction.  than  light.  found t h a t the a c t i o n o f  urease was  o f the ensyme, the v a r i o u s  was  dependent on the  dilution  i m p u r i t i e s p r e s e n t and  upon the  Radium r a d i a t i o n s were a l s o found t o be i n j u r i o u s  t o r e n n e t but t o have a s t i m u l a t i v e e f f e c t on d i a s t a s e . X - r a y s a l s o have v a r i e d e f f e c t s on enzymes, t h e d i a s t a t i c a c t i v i t y o f u r i n e , serum and w h i l e t h a t o f p e p s i n and H.  various f l u i d s being  amylase was  3. F u l l e r  considerably a l t e r e d .  (7) experimented w i t h  t i s s u e i n j u r e d by u l t r a v i o l e t c a t a l a s e anc  plant  r a d i a t i o n In respect  d i a s t a s e a c t i v i t y and  o f t h e s e enssyaes a c t u a l l y i n c r e a s e d  found t h a t t h e  to activity  to a considerable  i n t i s s u e s exposed t o d e s t r u c t i v e wave l e n g t h s and t h e r e was  unchanged  extent  that  no e v i d e n c e whatsoever of a d i m i n n i t i o n o f ensyme  B9.  a c t i v i t y , a c o n d i t i o n c o n t r a r y to t h e r e s u l t s o f p r e v i o u s Investigators.  This anthor b e l i e v e s t h a t t h e i n j u r i o u s  effects of ultra violet  l i g h t must be t r a c e a b l e t o some  p h y s i o l o g i c a l A«; angeaent o t h e r t h a t t h e i n a o t i v a t i o n o f enaymea. I t would s e e s t h a t v a r i o u s forms o f r a d i a n t energy have v a r y i n g e f f e c t s on enaymes from d iff©rent s o u r c e s and the work r e p o r t e d h e r e i s an attempt t o f i n d some c o r r e l a t i o n between t h e e f f e c t o f l i g h t on p l a n t emsymes and on a n i m a l ensymes i n t h e hope t h a t sons d e f i n i t e c l a s s i f i c a t i o n might be found and t h a t some conn e c t i o n might be e s t a b l i s h e d between t h e s e r e s u l t s and t h e r e s u l t s p r e v i o u s l y reported  on t h e e f f e c t on g r o w t h and r e -  production.  W2?MmmTAi WORK;. PART 3».  E f f e c t o f R a d i a n t Energy on D i e s t a s e .  PARS I I . E f f e c t o f R a d i a n t Bnergy on Oxidase A c t i v i t y o f Potato e x t r a c t . ^  X* The  most w i d e l y d i s t r i b u t e d ensymes t h a t c o n t r o l  t h e b r e a k i n g down Of t h e h i g h e r c a r b o h y d r a t e s a r e t h e Diastases ( Amylases).  These ensymes break down c a r b o h y d r a t e s  t o an end product o f m a l t o s e w i t h t h e i n t e r m e d i a t e o f e r y t h r o and a o h r o o - d e x t r l n s .  production  I n t h e s e experiments t h e  3©  a n i m a l amylase need was P t y a l i n , t h e d i a a t a t i c enayme p r e s ent i n human s a l i v a ,  t h e p l a n t enzyme used was rfe.lt  D i a s t a s e , fro® g e m i n a t e d  barley.  f h e d i v e r g e n c e I n r e s u l t s o b t a i n e d d u r i n g the b r e a k i n g down o f s t a r c h by d i a s t a s e o f v a r i o u s o r i g i n s has l e d t o t h e b e l i e f I n the p r e s e n c e o f two enzymes, a s a c c h a r o g e n i o enzyme and a s t a r c h l i q u i f y i n g ©n&yae.  Sherman  and So h i ©singer ( 3 3 ) and K e n d a l and Sherman (?.l) have demonstrated t h a t t h e l i q u i f y i n g a c t i o n i s predominant i n p a n c r e a t i c amylase and t h e s a c c h a r i f y i n g i n malt d i a s t a s e , and have a l s o shown t h a t t h e l i q u i f y i n g enzyme i s predominant i n s a l i v a r y d i a s t a s e .  These workers found t h a t t h e  p r o p o r t i o n o f t h e two a c t i o n s was u s u a l l y c o n s t a n t f o r each p a r t i c u l a r ensyme a l t h o u g h g r e a t d i f f e r e n c e s i n p r o p o r t i o n were n o t i c e a b l e i n en^ymea o f v a r i e d s o u r c e s , t h i s d i f f e r e n c e being ascribed  " t o the presence of complements  or a c t i v a t o r s which are required  f o r s p l i t t i n g up o f c e r t a i n  d e f i n i t e a t o m i c groups'*. Sow i t was thought t h a t l i g h t , h a v i n g a d e f i n i t e e f f e c t on ensyme a c t i o n , might have a v a r y i n g e f f e c t on t h e two enzymes p r e s e n t I n D i a s t a s e , a c t i n g as an i n h i b i t o r o r a c t i v a t o r o f the d i f f e r e n t g r o u p s , described  f h e e x p e r i m e n t s here  were t h e r e f o r e c a r r i e d out on the a n x i o l y t i c  a c t i o n o f d i a s t a s e as d i s t i n c t from t h e s a c c h a r o g e n i o a c t i o n , hut i t I s hoped i n t h e near f u t u r e t o a s c e r t a i n t h e e f f e e t on t h e s u g a r f o r m i n g ensyme and t o f i n d whether t h e d i f f e r e n c e s  31 are. l i m i t e d t o e i t h e r p l a n t o r a n i m a l amylase o r possessed  by b o t h .  HAfSRlAja 1. riialt D i a s t a s e .  .76 grms. of jAalt D i a s t a s e were  shaken f o r 16 minates i n £50 c e . d i s t i l l e d water and then f i l t e r e d .  S h i e s o l u t i o n p l u s two d r o p s o f t o l u e n e as  a p r e s e r v a t i v e was kept i n t h e d a r k i n a s t o p p e r e d b o t t l e . £.  Salivary diastase.  thus:40 °F.  Samples o f s a l i v a were o b t a i n e d  t h e mouth was washed o u t w i t h d i s t i l l e d w a t e r a t 80 c o . o f water was then h e l d i n t h e month f o r one  minute and ^ c o l l e c t e d •  f h i e was r e p e a t e d t w i c e ,  40 c c were t h e n shaken, f i l t e r e d u t i o n kept i n t h e d a r k ,  fh®  and. t h e r e s u l t i n g  sol-  i t was found t h a t t h e a c t i v i t y  o f t h e s o l u t i o n became l e s s on t h e t h i r d day so t h a t a f r e s h s o l u t i o n was made up e v e r y t h r e e days and was found t o be r e m a r k a b l y c o n s t a n t i n i t s a c t i v i t y ,  f h e achromio  p o i n t o f t h e s a l i v a was a s c e r t a i n e d and i f i t v a r i e d g r e a t l y from f o u r minutes adjustment was made o f t h e c o n c e n t r a t i o n o f the s o l u t i o n  t o o n t a i n an a chromic p o i n t  of constant value. 3.  S u b s t r a t e used.  C. P. s o l u b l e s t a r c h was used  throughout a l l t h e experiments. S o l u t i o n s o f s t a r c h were made up o f t h e f o l l o w i n g concentrations:-  0.1$, 0.8 ?4, 0.3 $  r  0.* ft, 0.75  1.0^,  32. 2.0$,  2.5f§, 3.0^, 4.0$, 5.0fl, I n t h e ease o f S a l i v a r y D i a s t a s e  £6 o.o. o f a  phosphate h n f f e r o f 6.7 pH were naed t o m a i n t a i n t h e pE maximum a t 6.7, w h i c h I s t h e opt imam pE f o r p t y a i i n a c t i v i t y . £5 c c .  o f Ifk N a C l were a l s o added as t h e a c t i o n  o f a n i m a l amylase has been proved t o be c o n d i t i o n e d  by  t h e presence o f n e u t r a l s a l t s i n q u a n t i t i e s v a r y i n g fro® .02^  t o 8.0 p. t h e G . l $ s t a r c h s o l u t i o n would t h e r e f o r e be  made up a s f o l l o w s : 4 25 25 146  c.o. 5 $ s t a r c h s o l u t i o n . c c . Suffer c c . 1£ l a C l c c D i s t i l l e d water.  f h e o t h e r s o l u t i o n s b e i n g made up w i t h t h e r e q u i r e d proportions  o f s t a r c h and d i s t i l l e d  water.  I n t h e ease o f t h e M a l t D i a s t a s e concentrations  t h e same  o f s t a r c h s o l u t i o n were lade up, but  t h e pH was 3 ept between 4.8 and 5.2 by means o f a p t h a l a t e hnffer.  The 0.1^ s t a r c h s o l u t i o n b e i n g made up as f o l l o w s 4 c c . 0> s t a r c h s o l u t i o n 25 c c B u f f e r 171 c c d i s t i l l e d w a t e r .  PROCTORS 3 c c  o f enzyme s o l u t i o n were p l a c e d  o f two q u a r t s t u b e s *  One tube was f i t t e d  i n each  into the s l i t  o f t h e monochromatic i l l u m i n a t o r o r i r r a d i a t e d d i r e c t l y i n f r o n t o f t h e q u a r t s mercury lamp and t h e o t h e r was  33. k e p t i n t h e d a r k as & c o n t r o l . In the  the case of s a l i v a r y diastase  i r r a d i a t e d so l o t i o n  1 c.c. o f  was d i l u t e d t o 100 c.c. w i t h d i s t -  i l l e d w a t e r and 1 c . c . o f t h i s s o l u t i o n p l a c e d i n each o f the  s e r i e s o f t e s t tubes c o n t a i n i n g 5 c . c . o f each o f t h e  starch solutions.  ?hus 0.01 c . c . o f ensyme was p r e s e n t i n  each t e s t t u b e p l u s 5 c-.c# s t a r c h s o l u t i o n , c e s s was r e p e a t e d f o r t h e c o n t r o l .  f h e same p r o -  One d r o p o f t o l u e n e  was added t o each t e s t t u b e , w h i c h was t h e n shal en, and p l a c e d i n an oven a t 38°-40°F. 3 d r o p s o f t h e i r r a d i a t e d malt d i a s t a s e were added by means o f a d r o p p i n g p i p p e t t t o t h e s t a r c h s e r i e s w i t h 8 d r o p s o f t o l u e n e p e r t e s t t u b e , shaken, and p l a c e d i n t h e oven at 38°-40°F.  Three drops o f t h e n o n - i r r a d i a t e d  c o n t r o l a l s o being added t o a n o t h e r s e r i e s and t h a t i n t u r n p l a c e d i n t h e oven. At  stated  i n t e r v a l s o f t i m e one d r o p was t a k e n  f r o m each o f t h e s e r i e s o f t e s t t u b e s , i r r a d i a t e d and c o n t r o l , p l a c e d on a g l a s s p l a t e , u s i n g a d r o p p i n g p i p e t t e , and one d r o p o f 1/10 I o d i n e added,  fhe f i r s t  appearance  of r e d c o l o u r a t i o n , d e n o t i n g c o n v e r s i o n o f s t a r c h t o d e x t r i n , was noted and t h e number o f t h e t e s t tube r e corded .  34.  BSSTOta. Sailvary 1«  3?oo e f f e c t o f f a l l  Maatase. i l l u m i n a t i o n from a q u a r t s  1««*P on t h e a m g l o l y t i c a c t i o n o f  a) ,  mercury  saliva.  f b r e e e.e. o f ensyme were i r r a d i a t e d  at v a r i o u s  d i s t a n c e s f r o m a q u a r t s mercury lamp f o r a p e r i o d o f one hour and t h e a e h r o a i c { f a b l e 6.)  point of the s o l u t i o n ascertained.  These r e s u l t s show t h a t t h e a c t i v i t y o f t h e  ensyme i s g r e a t l y reduced by t h e f i c t i o n o f l i g h t i n d i r e c t p r o p o r t i o n t o t h e d i s t a n c e from t h e b) .  source.  f h r e e c . c . samples o f s a l i v a r y d i a s t a s e were i r -  r a d i a t e d f o r one hour at d i s t a n c e s o f 10, 7, 5 and 3 erne, from the source of l i g h t . radiated  The  e f f e c t of t h e v a r i o u s i r -  ensyme s o l u t i o n s on t h e s e r i e s of s t a r c h t u b e s a s  p r e v i o u s l y d e s c r i b e d was a s c e r t a i n e d a t varj-ing p e r i o d s t i m e , and a c o m p a r i s o n made -vith t h e c o n t r o l .  of  ( f a b l e 7,  P l a t e 8). I t w i l l be seen t h a t the f u l l l i g h t mercury a r e lamp has a d e c i d e d  from a  i n h i b i t o r y a c t i o n on the  a c t i v i t y of s a l i v a r y d i a s t a s e , e v i d e n t l y i n proportion t o the i n t e n s i t y of t h e l i g h t ,  fhe r i s e  i n t e m p e r a t u r e o f the ex-  t r a c t s so i r r a d i a t e d was n e g l i g a b l e as compered w i t h t h e c o n t r o l except i n the case o f i r r a d i a t i o n a t a d i s t a n c e o f 3 cms.  from t h e s o u r f e o f l i g h t when the t e m p e r a t u r e was  r a i s e d about 3°C. above room t e m p e r a t u r e ,  f h e almost t o t a l  35 ffA3&3 6. fhe  e f f e c t o f I r r a d i a t i o n on t h e a c h r o a l o p o i n t of s a l i v a . ~" D i s t a n c e from L i g h t Soarce 30 80 10 7 5 3  Ac hr p a l e  cms. mm* cms. cms. ems. cms.  4 6 IB 36 64 Ho  Point.  tains. mine. rains. mine. tains. end p o i n t .  fASLE'-7. f K l e f f e c t o f f u l l I l l u m i n a t i o n from a g j i a r t u mercury lamp on t h e a c t i v i t y o f S a l i v a r y D i a s t a s e . A c t i v i t y o f ensyrae expressed as amount o f star«fh reduced* ( P e r c e n t a g e c o n c e n t r a t i o n of s o l u t i o n J . Hours.  I S  3  4  6  6  7  8  9  10  84  48  96  -  -  E.O  -  -  - 8.5 3.0 4.0 5.0  .75 1.0 -  -  1.5  -  - E.O 8.5 3.0 4.0  -  - 1.5 2.0 £.6 3.0  -  - .75 1.0 1.5 8.0  Control .5 10 cms..5  1.0 1.5  7 cms. *£  -  .6 »  -  1.0  5 cms. ..£  -  .3 -  -  .5  -  PIAfB 8  B f f e c t of f n l l i l l u m i n a t i o n f r o g a g n a r t s aero o rj lamp on t h e a m y l o l y T i c a c t i o n o f s a l i v a ana o f malt d i a s t a s e .  Q r d i n a t e s . Ski^yme a c t i v i t y expressed percentage concentration o f s t a r c h s o l at i o n reduced. A b s c i s s a e , SDime a f t e r c e s s a t i o n o f Irradiation. A.  B f f e o t on S a l i v a r y D i a s t a s e .  B.  S f f e c t on M a l t  Diastase.  V  36. i n a c t i v a t i o n o f the mnjme so i r r a d i a t e d may p a r t l y due  he t h e r e f o r e  t o t h e d e s t r u c t i v e a c t i o n of the wave l e n g t h s  o f the l o n g e r p a r t o f t h e It  spectrum.  i s o f i n t e r e s t t o n o t e t h a t an experiment  c a r r i e d out t o a s c e r t a i n the approximate e f f e c t of  was  full  i l l u m i n a t i o n on t h e amount o f r e d u c i n g sugar formed and e f f e c t was  proved t o he d e c i d e d l y i n h i b i t o r y i n n a t u r e *  B. f h e s p e c i fl e i f as t as®. '  e f f e c t s o f Monochromatic L i g h t on S a l i v a r y  f a k i n g t h e s e r e s u l t s i n c l a s s e s we C1*8B i«  the  l a v e l e n g t h s h a v i n g no  have:-  effect.  t h e s e i n c l u d e 6152 A°, 5819  A° and  3821 A ° , w h i c h  e x e r t no measurable e f f e c t on the a c t i v i t y o f t h e enzyme. C l a s s 2*  l a v e lengths having a s t i m u l a t i n g e f f e c t . 4359 A  0  and 4078 A  0  a p p e a r to have a  slight  s t i m u l a t o r y e f f e c t , which i s l o s t , however, a f t e r some h o u r s . 3663 A  on the o t h e r hand e x e r t s a s l i g h t i n h i b i t o r y e f f e c t  0  at f i r s t ,  f o l l o w e d by a s t i m u l a t i o n w h i c h i s maintained  through-  out the experiment. C l a s s .3.  Save l e n g t h s c a u s i n g a r e t a r d a t i o n In a c t i o n . 5461 A°* 4916  A°, 3132A w i t h a l l l i n e s of 0  s h o r t e r wave l e n g t h possess a decided which i s maintained 3022 A  0  inhibitory  throughout t h e p e r i o d .  the i n h i b i t i o n i s - n l y i n i t i a l and  t a k e s p l a c e by t h e c o m p l e t i o n  effect,  In the c a s e of fall  of the experiment.  recovery  37. g a b l e 8. The s p e c i f i c e f f e c t s o f Monochromatic L i g h t on P a l l y Diastase. A c t i v i t y o f t h e enayme expressed as amount o f s t a r c h reduced, p e r c e n t a g e c o n c e n t r a t i o n o f s o l u t i o n * Readings  a f t e r t w e n t y - f o u r hours  Hours.  1  Control 616EA  0  •5  3 1.0  4  1*5 -  So e f f e c t .. - * » » * #  5819AO f o e f f e c t ..  Aft*  *  5461A° .3  . 5 .75 -  4916A .3  •5  0  1.5  4078A .75 1.0  1.5  0  3821A  1.0  3132A .3  .3  3G22A .3  .5  2967A  0  .3  .5  2804A  0  .3  .5  8700A .5  .5  2535A  .5  0  0  0  0  .3  7  8  9  10  2.0  -  -  _  2/S 5.0  * «  1.0  1.5  -  -  -  1.5  —  - -  - -  2.0 » ** •  - .5 - .75 - 1.0 .75 - - 1.0 - - .75 - - 1.0 - .75 1.0 .75 1.0 .75 1.0 — 1.5 1*5  84  48  96  4.0 5.0  * * • *••••»«••• * * * • « • * • «  - £.0  l o e f f e c t ..  3663AO .5  6  * * * **• ***  .75 1.0 -  4359A .75 1.0 0  0  5  Irradiation.  - 2.0  _  -  2.5  3.0 4.0  2.0 2.5  3.0 4.0  8.0 3.0  4.0 5.0  2.0 3.0  4.0 5.0  4 ***** a**  3.0 4.0  6.0  -  8.0  3.0 4.0  -  3.0  4.0 4.0  -  2.6  3.0 4.0  -  £.5  3.0 4.0  -  8.0  3.0 4.0  -  2.0  8.5 3.0  38 • Besalls 1*  fhe  M a l t Diastase*. effeet of f u l l Illumination  lamp on t h e a c t i v i t y of malt  from a g u a r t a mercury  diastase*  A decided I n h i b i t o r y e f f e c t i s recorded.,  increas-  i n g w i t h t h e i n t e n s i t y of t h e l i g h t , the g r e a t e s t I n h i b i t i o n being found when the e o l a t i o n s were i r r a d i a t e d a t 3 ems. d i s t a n t from the s o u r c e o f 2.  The  5461  f Plate  8)  s p e c i f i c e f f e c t s o f monochromatic l i g h t on  a c t i v i t y o f malt o  ight.  diastase*  i t was foand t h a t o n l y f o u r wave l e n g t h s , i * e . , _  A , 4916  A  , 31'2  A  0  and  3022 A  had  0  any  t h e d l a s t a t i c a c t i o n o f the malt s o l u t i o n . e f f e c t i n the s h o r t e r the  f a c t t h a t the  wave l e n g t h s may  i n t e n s i t y o f the  s u f f i c i e n t t o e f f e c t any i t must be noted t h a t 3132 l i n e s o f v e r y low 5461  A, 0  the  and  on  lack  of  The  be due  incident  effect  largely  l i g h t was  change i n ensyae a c t i o n , A , 0  3022 A  0  and  4916  A  not  although are a l l  0  i n t e n s i t y , e s p e c i a l l y as compared  to  a l l exert a decidedly i n h i b i t o r y a c t i o n ,  i n g to the f a c t t h a t i n t e n s i t y Is  not  the  to  point-  controlling  f a c t o r i n t h i s o ase*  COfeCPARISOy OF HKBgXffS» 1. diastase  I n both c a s e s , th«t of s a l i v a r y and  malt  f u l l i l l u m i n a t i o n from a q u a r t s mercury lamp  has  a d e c i d e d i n h i b i t o r y e f f e c t on the a c t i v i t y o f t h e s e ensymee. I n the c a s e o f s a l i v a the most i n t e n s e i l l u m i n a t i o n used  39. TARLK 9. g f f e o t o f f a l l I l l u m i n a t i o n from a q u a r t * mercury lamp on the a c t i v i t y of H a l t  Piastaae.  Kusyme a c t i v i t y expressed a s amount o f s t a r c h r e d u c e d . Headings a f t e r t w e n t y - f o u r hours B^ys.  1 8  irradiation.  3  4  6  6  ?  8  9  10  1.6 8.0 10 ems.  -  8.5  -  -  3.0 -  -  4.0  1.0 1.6  -  -  -  8«6  -  -  3.0  7 cms 1.0 1.6  -  -  8.0  -  -  - 8.5 3.0  5 cms .75 1.0  -  -  1.5  -  -  -  -  2.5  fl cms•.3  -  .76  -  1.0  -  -  -  1.5  11  IE  dontreX  .5  -  3.0  10. S p e c i f i c e f f e c t s o f i*ioaocbroaatlo l i g h t on t h e a c t i v i t y of M a l t  Diastase.  Control 1.5  8.0 -  2.5  -  -  3.0  4.0  5461A  0  1.5  -  -  2.0  -  -  8.5 - -  3.0  4916A  0  1.5  -  -  -  8.0  -  8.5 - -  3.0  -  4.0  3138A  0  1.6  - 8.0  -  -  -  8.6 - -  3.0  -  4.0  2.0  -  -  3.0  -  4.0  3088A° 1.6  -  -  -  2.5  40. ( Seas, d i s t a n t ) has an a l m o s t c o m p l e t e l y  paralysing effect  i n c o n t r a s t to m a l t d i a s t a s e , which shows g r a d u a l from t h e e f f e c t s of i r r a d i a t i o n a t t h a t d i s t a n c e  recovery (Plate 8).  I t must be rembered, however, t h a t t h e normal a c t i v i t y of t h e two amylases i s v e r y d i f f e r e n t , the s t a r c h l i q u i f y i n g ensyme i n s a l i v a b e i n g many times more r a p i d i n I t s a c t i o n t h a n t h e l i q u i f y i n g p o r t i o n i n malt d i a s t a s e , f h e s e r e s u l t s may be i n p a r t a t t r i b u t a b l e t o t h i s d i f f e r e n c e i n a c t i v i t y , t h e l e a s t a c t i v e ensyme being 2.  the l e a s t a f f e c t e d .  fhe r e s u l t s from I l l u m i n a t i o n w i t h mono-  c h r o m a t i c l i g h t a r e i n t e r e s t i n g i n t h a t f o u r l i n e s , 5461 A , 0  4916 A , 3132 A 0  0  and 3022 A  0  i n a l l cases exert a decidedly  I n h i b i t o r y a c t i o n on the a c t i v i t y o f t h e ensymes.  fhat t h i s  i s due s o l e l y t o wave l e n g t h and not t o i n t e n s i t y can be o c l e a r l y seen by comparing the r e l a t i v e i n t e n s i t i e s o f 8461 A and  4916 A , l i n e s w h i c h e x e r t , e s p e c i a l l y i n t h e c a s e o f 0  M a l t d i a s t a s e , p r a c t i c a l l y t h e same amount o f . I n h i b i t i o n w i t h very unequal i n t e n s i t i e s . fhewave l e n g t h s t h a t cause s t i m u l a t i o n i n s a l i v a r y d i a s t a s e have no e f f e c t on M a l t d i a s t a s e , i . e . , 4359 A , 0  4078 A° and 3663 A ° . I n s a l i v a r y d i a s t a s e a l l wave l e n g t h s t h a n 3022 A  0  exert a decidedly  shorter  i n h i b i t o r y e f f e c t on the  ensyme a c t i v i t y but have no e f f e c t on t h a t of malt d i a s t a s e .  PMfE  9  Comparison o f the e f f e c t o f aonochrotaat lo l i g h t on t h e growth o f Baramoeoinm oaudatnm ana on t h e a c t i v i t y o f a n i m a l diastase":  Paramoecium - i n c r e a s e o r d e c r e a s e i n nranher o f e e l l s a f t e r t w e n t y - f o u r hours i r r a d i a t i o n over the c o n t r o l count. Ensyme a c t i v i t y * - expressed as p e r c e n t a g e conc e n t r a t i o n o f s t a r c h s o l u t i o n reduced a t p e r i o d s o f t h r e e and t w e n t y - f o u r hours a f t e r i r r a d i a t i o n .  utt siii  3t5b  mi mi M M  «7z  mi mi i%i  WK  mn uis zos'tK  41. Comparison o f t h e e f f e c t of n i t r e v i o l e t 1ight on growth w i t h the  e f f e c t on ensyme a c t i o n . 1.  Comparison o f the  e f f e c t of Monochromatic  l i g h t on growth of a n i m a l p r o t o p l a s m end o f animal i i a a t a s e  . ( Plate  en t h e  activity  9.)  f h e r e appears to be some c o r r e l a t i o n between r e s u l t s o f t h e s e e x p e r i m e n t s . f%e  l i n e c a u s i n g the  the  greatest o  stimulation  o f growth i n Paramoeniurn, i . e . ,  t h e maximum s t i m u l a t o r y diastase.  , shows  e f f e c t on the a c t i v i t y of s a l i v a r y  Those l i n e s t h a t  only s l i g h t l y stimulate  appear t o hr-ve no e f f e c t on the a c t i v i t y of the She  A  s h o r t e r wave l e n g t h s , c a u s i n g an  growth  enzyme.  i n h i b i t i o n of growth  i n Paramoeoina, e x e r t a decided i n a c t i v a t i n g e f f e c t on eawyme. 4078 A , 0  e f f e c t the  c a u s i n g I n h i b i t i o n i n growth does not  ensyme a c t i v i t y and  g r o w t h , has  an  31?»E A ,  having no  effect  l i n ^ s g i v i n g divergent r e s u l t s  a c o r r e l a t i o n between growth r a t e and  a t i c a c t i v i t y under the c o n d i t i o n s { Plate  0  i n a c t i v a t i n g e f f e c t on ensyme a c t i o n ,  a p a r t from t h e s e two appears to be  the  on  but there d iast-  of t h e s e e x p e r i m e n t s .  9.) 2.  Comparison o f the  on the growth of p l a n t  e f f e c t of monochromatic l i g f c t  protoplasm and  on the a c t i v i t y of  plant  d iastase. There seems to be no e v i d e n t c o r r e l a t i o n between the r e s u l t s obtained f o r malt d i a s t a s e  and  t h o s e on  the  42. growth mi 4916  sporulation of Colletotrichraa,  although  A° and 3022 A® cause i n h i b i t i o n i n a l l o a s e s .  This  l a c k o f c o r r e l a t i o n i s perhaps due t o t h e f a c t that malt d i a s t a s e i s an ensyme from a p l a n t c o n t a i n i n g c h l o r p h y l l and C o l l e t o t r i e h u m i s a f u n g u s ,  f h e r e may be a d i f f e r e n c e  between t h e s e n s i t i v i t y of enzymes i n p h o t o s y n t h e t i c and  plants  i n t h o s e l a c k i n g c h l o r o p h y l l . E x p e r i m e n t s a r e t o be  undertaken on enaymes obtained  from f u n g i i n o r d e r t o  c l a r i f y this point. S u f f i c i e n t d a t a has not y e t been obtained t o e n a b l e any d e f i n i t e s t a t e m e n t t o be mode, but t h e r e s u l t s so f a r p o i n t t o t h e f a c t t h a t p l a n t d i a s t a s e i s l e s s s e n s i t i v e t o l i g h t o f v a r i o u s wave l e n g t h s than a n i m a l d i a s t a s e .  43 f h e e f f e c t o f R a d i a n t Energy OR t h e Oxidase  B&Rf, B.  a c t i v i t y o f Potato J u i c e * P o t a t o e x t r a c t was used i n t h e f o l l o w i n g experiments t o determine  the e f f e c t of n l t r a v i o l e t  light  and monochromatic l i g h t on o x i d a s e a c t i v i t y . f h e method o f measuring  the oxidase a c t i v i t y  was based on t h a t o u t l i n e d by G u t h r i e ( 1 2 . )  Be noted  t h a t p o t a t o j u i c e c o n t a i n e d a substance t h a t might be t i t r a t e d with iodine i n acid s o l u t i o n ( t r i c h l o r a c e t i c a c i d ) and t h a t t h i s t i t r a t i o n v a l u e decreased  on exposure  t o a i r . However, i n some cases t h e t i t r a t i o n was found Jre be v e r y much Increased and i t was b e l i e v e d t o be due t o a low c o n t e n t o f o x i d a s e , t h e "substance r e s p o n s i b l e f o r the i o d i n e r e a c t i o n not being oxidised cess o f e x t r a c t i o n f o r t h i s reason".  In the pro-  G u t h r i e found  t h a t g l u c o s e warmed w i t h d i l u t e HaOE c o n t a i n e d a c a r b o hydrate d e r i v a t i v e capable solution.  f reducing Iodine In acid  T h i s t i t r a t i o n decreased  on exposure  and t h e o x i d a t i o n was c a t a l y s e d by p o t a t o j u i c e .  to a i r I t would  t h e r e f o r e be p o s s i b l e t o measure t h e a c t i v i t y o f t h e oxi d a s e i n t h e p o t a t o j u i c e -y t h e amount o f i o d i n e r e duced by j u i c e and s u b s t r a t e . :HB?P0PS. 1.  Preparation of substrate  40 g r a s . o f g l u c o s e were d i s s o l v e d  v  i n 400 c , c . o f Normal  sodium h y d r o x i d e , placed i n a 500 c.e. f l a s k and immersed  44. i n a water-bath was  a t 80°C f o r IS m i n u t e s .  The  solution  removed and n e u t r a l i s e d a t once by adding 10 c . c . of  85$> p h o s p h o r i c was added and m i x t u r e was  acid.  2b grins, o f d e c o l o u r i z i n g c h a r c o a l  t h e m i x t u r e a l l o w e d t o stand o v e r n i g h t ,  t h e n f i l t e r e d and  a n o t h e r 25 grms. of c h a r -  c o a l added and  a l l o w e d t o stand f o r 15 minutes and  filtered,  pE was  H/10  fhe  fhe  then a d j u s t e d t o 6.5  again  by means o f  sodium h y d r t x i d e o r h y d r o c h l o r i c a c i d . 2.  P r e p a r a t i o n and  i r r a d i a t i o n o f ensyme e x t r a c t *  An e x t r a c t o f p o t a t o j u i c e c o n t a i n i n g the ensyme was  ob-  t a i n e d by g r i n d i n g up a measured amount o f p o t a t o and s q u e e z i n g the ground m a t e r i a l through cheese c l o t h w i t h a p o t a t o masher.  3 c c . o f the e x t r a c t so obtained  p l a c e d i n a q u a r t s c o n t a i n e r , stoppered and  either irradiated  was  w i t h cotton wool,  by t h e f u l l i l l u m i n a t i o n from a  q u a r t s mercury lamp a r p l a c e d i n the s l o t of the monochromatic  i l l u m i n a t o r , f o r v a r y i n g p e r i o d s o f time. 3.  80 c c  of oxidase a c t i v i t y .  o f the s u b s t r a t e d i l u t e d  w a t e r was B.  Meaanreaerit  1:1 w i t h d i s t i l ' ed  p l a c e d i n each o f two a e r a t i o n t u b e s , A and  fO A was added 3 c c  o f p o t a t o e x t r a c t which had  been kept In the dark d u r i n g t h e i r r a d i a t i o n p e r i o d t o a c t as a c o n t r o l ,  f o B. was added 3 c . c . of i r r a d i a t e d  e x t r a c t . 5 drops of p a r a f f i n o i l were added t o each t o a c t as a foam b r e a k e r , and both A and  3 were a e r a t e d  f o r one hour w i t h <?qual c u r r e n t s of a i r . At t h e end  45. o f one h o u r 5 c.o.  from each A and  3 were washed i n t o  each o f f o u r f l a s k s c o n t a i n i n g 5 c c . a c e t i c a c i d and  10 c c .  of 10%  o f I o d i n e . { N/50  trichlor-  I i n 1/10  KI).  A f t e r f i v e and t e n m i n u t e s , two f l a i l s , c o n t r o l and i r r a d i a t e d r e s p e c t i v e l y were t i t r a t e d w i t h S/100  Sodium  t h i o s o l p h a t e u s i n g 1 c.e.  an  indicator, and  B was  o f 1 % s t a r c h p a s t e as  f h e d i f f e r e n c e between the t i t r a t i o n s of A used as a measure o f t h e d i f f e r e n c e between  the o x i d a s e a c t i v i t y o f t h e c o n t r o l ana  the  irradiated  sample, a l a r g e r t i t r a t i o n i n B ( i r r a d i a t e d ) d e n o t i n g a g r e a t e r o x i d a s e a c t i v i t y as compared t o t h e c o n t r o l ( A ) , and  vica versa.  TOKHUKSf  I  I r r a d i a t i o n w i t h the f u l l  I l l u m i n a t i o n from a quart?, mercury  lamp. 3 cc*  p o r t i o n s o f p o t a t o e x t r a c t were i r r a d i a t e d :1*  i n glass containers.  E.  i n quartz  containers,  a t a d i s t a n c e o f 10 cms.  f r o . i t h e .source o f  1  ight, for  v a r y i n g p e r i o d s o f t i m e , t h e t e m p e r a t u r e being £2°C, room t e m p e r a t u r e Results. ( fable  1-  I r r a d i a t i o n In g l a s s c o n t a i n e r s .  11.) Ip I s p r o b a b l e t h a t the g l a s s of t h e s e c o n t a i n e r s o  a b s o r b s a l l wave l e n g t h s any  1  ess  t h a n about 3300 A , so t h a t  e f f e c t on the o x i d a s e a c t i v i t y would be caused by  the  46 TABLE I I  KFFKCg of i l l u m i n a t i o n from t h e rays of & q u a r t g merc u r y lamp on o x i d a s e a c t i v i t y of potatoIWlceT D i s t a n c e from s o u r c e 10 cms. O x i d a s e a c t i v i t y expressed a s eonfcr©1 t i t r a t i ons. A.  Irradiated  i n glass c o n t a i n e r s *  Time o f Irrad iation 1 hour  i n c r e a s e or d e c r e a s e over  Readings a f t e r 6 minutes  Readings a f t e r 10 m i n u t e s .  -5.7  —4.6  £ hours  1.5  5.7  3 hears  10.3  10.6  4 hours  13.6  13. 1  5 hours  5.i  4.3  6 hours  4.6  7 hours  3.6  4.0 3.5  I r r a d i a t e d I n Quart s cent a i n e r s . 1 hour  -4.£  - £«1  £ hours  -13.8  -10.8  3 hours  - 6.£  - 3.9  4 hours  - 4.8  - 3.7  5 hours  - 3.5  - 3.8  6 hours  - 3.4  - 3.8  7 hours  - 3.£  -3.6  PMfB  10.  B f f e c t o f i l l u m i n a t i o n from a raercnrv are lamp on the o x i d a s e a c t i v i t y o f potato extract.  A.  I l l u m i n a t i o n through a g l a s s  3.  I l l u m i n a t i o n through, a q u a r t s  container. container.  Oxidase a c t i v i t y expressed as p e r c e n t a g e i n c r e a s e o r d e c r e a s e over the c o n t r o l t i t rat ions.  47  r i s i b l e and some o f t h e n e a r u l t r a v i o l e t wave l e n g t h s . I t would seem from P l a t e 10 t h a t t h e ensyme a c t i v i t y was decreased a t f i r s t , but a f ~ e r one hour's I r r a d i a t i o n I t was d i s t i n c t l y i n c r e a s e d a s compared t o t h e c o n t r o l a t t h a t t i m e , and t h a t a f t e r seven hours i r r a d i a t i o n was s t i l l a c t i v e than t h e c o n t r o l e x t r a c t .  more  She maximum a c t i v i t y  b e i n g shown a f t e r a p p r o x i m a t e l y t h r e e hours i r r a d i a t i o n . £.  I r r a d i a t i o n i n quarts containers.  These q u a r t s v e s s e l s do not absorb any o f t h e f a r u l t r a v i o l e t l i n e s u n t i l around 2500 A . 0  I t w i l l be seen t h a t  at a l l p e r i o d s o f i r r a d i a t i o n t h e o x i d a s e a c t i v i t y was d e c r e a s e d , r e a c h i n g a minimum a t around t h e two hours and then g r a d u a l l y r e t u r n i n g nearer t o the c o n t r o l .  But a f t e r  a p e r i o d o f e i g h t e e n hours exposure the d e c r e a s e was v e r y marked*  I t would see i from t h e above r e s u l t s t h a t t h e f a r  u l t r a v i o l e t l i n e s have an i n h i b i t o r y e f f e c t on o x i d a s e a c t i v i t y s u f f i c i e n t t o c o u n t e r a c t any s t i m u l a t i o n caused by t h e wave l e n g t h s o f t h e v i s i b l e and near u l t r a fhe  violet,  p o i n t o f maximum e f f e c t seemed t o be around 3-4 hours  w i t h t h e g l a s s and two t o t h r e e hours w i t h the q u a r t s containers. KEP?SRM5H? 2.  E f f e c t o f monochromatic l i g h t on o x i d a s e  a c t i v i t y of potato e x t r a c t . 3 c . c . o f p o t a t o e x t r a c t were p l a c e d  i n a quart*  c o n t a i n e r i n t h e s l o t o f a monochromatic i l l u m i n a t o r f o r  48  p e r i o d s o f t h r e e and i n some eases e i g h t e e n hours and s u b j e c t e d t o t h e v a r i o u s monochromatic l i n e s o f t h e mercury a r c s p e c t r u m .  3 c.e. of t h e same e x t r a c t were  p l a c e d I n t h e d a r k , a t t h e same t e m p e r a t u r e , t e a c t as a control. E M S . C l a s s 1. o 6152 A  Stimulation causes a c o n s i d e r a b l e i n c r e a s e i n a c t i v i t y w h i c h ,  i s however, decreased a f t e r 18 hour a i r r a d i a t i o n due proba b l y t o t h e e q u i l i b r i u m b e i n g reached m r e q u i c i l y t h a n in the c o n t r o l .  5819 A  0  causes an I n c r e a s e i n a c t i v i t y  m a i n t a i n e d throughout t h e e i g h t e e n h o u r s , and 5461 A o o and 4960 A r e s u l t i n t h e same i n c r e a s e i n a c t i v i t y . 4359 A 0  and 3881 A  0  cause o n l y a s l i g h t  increase i n a c t i v i t y  main-  t a i n e d throughout t h e whole p e r i o d . C l a s s 11. 4359 A , 4078 A 0  0  seen t o have but a l i g h t e f f e c t , w h i c h may  be covered by e x p e r i m e n t a l e r r o r , but I n no c a s e a r e t h e effects entirely negative. g l a s s 111. 3132 A ° , 3022 A ° , 2804 A  0  and 2700 A  0  cause decided and  c o n t i n u e d d e c r e a s e i n a c t i v i t y . 2535 A , 2064 A ° , both 0  approaching the l i m i t s of t r a n s i t i o n o f q u i r t s g l a s s , cause s l i g h t but c o n t i n u e d d e c r e a s e i n a c t i v i t y .  49 fABLE I B . Specifio  e f f e c t s of monochromatic 1 i g h t •  O x i d a s e a c t i v i t y expressed as $> i n c r e a s e or d e c r e a s e over control titrations. Wave l e n g t h in A 0  Time Irrad.  Headings a f t e r 5 minutes.  Read i i igs a f t e r 10 m i n u t e s .  6158  3 hrs. 18 hours  3.3 - 2.2  1.5 -1.4  5819  3 hours 18 hours  1.4 .6  2.1  5461  3 hours 18 hours  1. 3 8.9  0 5.2  4960  3 hours 18 hours  1.3 8.6  2.0  4359  3 hours  .6  0  4078  3 hours  .6  .6  3881  3 hours  3132 3082  1.  1.4  .6  8  .6  3 hours 18 hours  -1.3 -2.4  -1.3 -8.4  3 hours 18 h o u r s  -3.7  .65 —11.3  2804  3 hours  -2.4  -2*5  2700  3 hours  -3.0  -3.0  2535  3 hours  -1.8  - .6  £054  3 hours  -1.6  - .5  59. .•piscpssioiF OF  nmmxa.  From t h e r e s u l t s j u s t r e c o r d e d i t would s e e n t h a t r i s i b l e and u l t r a v i o l e t l i g h t have an a p p r e c i a b l e e f f e c t on the o x i d a s e a c t i v i t y o f p o t a t o e x t r a c t under t h e cond i t i o n s o f these experiments. V i s i b l e l i g h t , especially the yellow portion o f t h e s p e c t r u m , c a u s e s a s t i m u l a t i o n , a l t h o u g h the t r a n s i t i o n l i n e s around 4078 A activity,  0  show a s l i g h t d e c r e a s e i n  f h e near u l t r a v i o l e t l i n e s cause a d e c r e a s e  i n a c t i v i t y w h i c h becomes g r e a t e r a s t h e f a r u l t r a  violet  i s approached. f h e t o t a l e f f e c t o f t h e l i n e s o f t h e -alercury A r c Speotrma c a u s i n g d e c r e a s e i n a c t i v i t y seems t o be g r e a t e r t h a n t h a t caused by t h e s t i m u l a t i n g l i n e s , as i s shown when i r r a d i a t i o n i s c a r r i e d out i n g l a s s as compared t o quarts containers.  I n t h e case, o f t h e q u a r t s c o n t a i n e r s ,  w h i c h do not a b s o r b l i g h t l o n g e r t h a n about 8 6 0 C iA°, cont i n u o u s i n h i b i t i o n i s r e p o r t e d , whereas when g l a s s cont a i n e r s , c u t t i n g out some o f t h n e a r and a l l o f the f a r u l t r a v i o l e t , a r e used, s t i m u l a t i o n i s found t o o c c u r i n p r a c t i c a l l y a i l cases.  SlftMABT OK BKaOLfS* 1. by  f h e a c t i v i t y o f s a l i v a r y and malt d i a s t a s e i s r e i a o M the a c t i o n o f l i g h t from a mercury aro lamp i n p r o p -  ortion to the intensity. 2.  f h e e f f e c t s of monochromatic l i g h t on t h e a c t i v i t y of  s a l i v a r y d i a s t a s e arc? o f two k i n d s , s t i m u l a t o r y and i n hibitory.  4359 A , 0  4078 A  0  and 3663 A  0  exerting stim-  u l a t i o n and 5461 A°, 4916 A ° , 3132 A° and 3022 A° e r t i n g an i n h i b i t o r y 3.  ex-  effect.  F o u r l i n e s o n l y o f the mercury a r c spectrum e x e r t  any e f f e c t on t h e a c t i v i t y o f malt d i a s t a s e , i . e . , 5461 A , 0  4916 A ° , 3132 A  0  and  3022 A . 0  This  e f f e c t i s one  of i n h i b i t i o n . 4.  Under the c o n d i t i o n s o f these e x p e r i m e n t s t h e r e  appears t o be some c o r r e l a t i o n between the e f f e c t o f t h e monochromatic 3 in.-js o f the merenry a r c spectrum on t h e growth o f Paramoeclorn and on the a c t i v i t y o f s a l i v a r y d i a s t a s e ; t h o s e l i n e s c a u s i n g s t i m u l a t i o n i n growth i n most c a s e s c a u s i n g Inerea> e i n ensyme a c t i o n and  those  l i n e s c a u s i n g i n h i b i t i o n o f growth d e c r e a s i n g the a c t i v i t y o f the ensyme. 6.  f h e r e appears t o be no c o r r e l a t i o n between the/results  o b t a i n e d f o r malt d i a s t a s e and f o r growth o f Col? e t o t r i c h u m . 6.  f h e r e a r e i n d i c a t i o n s th- t under the c o n d i t i o n s o f  these experiments plant d i a s t a s e i s l e s s s e n s i t i v e t o l i g h t from the mercury a r c spectrum than i s a n i m a l d i a s t a s e .  52. 7.  f h e v i s i b l e and near u l t r a v i o l e t l i n e s of the mer-  c u r y a r c spectrum c a u s e an i n c r e a s e i n t h e o x i d a s e a c t i v i t y of potato e x t r a c t . 8.  f h e f a r n l t r a v i o l e t cause a d e c r e a s e  i n oxidase  a c t i v i t y of potato e x t r a c t . 9.  ftie t o t a l e f f e e t o f the l i n e s o f the mercury a r c  spectrum c a u s i n g i n h i b i t i o n o f o x i d a s e a c t i v i t y see;is t o be g r e a t e r than t h a t o f the l i n e s c a u s i n g i n c r e a s e in  activity  53.  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