University of Southern California Dissertations and Theses

A study of the histological distribution of glycogen in tumors of hypo-physectomized and non-hypophysectomized rats

(USC Thesis Other)

A study of the histological distribution of glycogen in tumors of hypo-physectomized and non-hypophysectomized rats

PDF

Download

Open document

Flip pages

Download a page range

Download transcript

Copy asset link

Request this asset

Transcript (if available)

Content A STUDY OF THE HISTOLOGICAL DISTRIBUTION OF GLYCOGEN IN TUM ORS OF HYPOPHYSECTOMIZED AND NON-HYPOPHYSECTOMIZED RATS A T h e s is P r e s e n te d to th e F a c u lty o f th e D epartm ent o f Zoology U n iv e rs ity o f S o u th e rn C a lif o r n ia I n P a r t i a l F u lf illm e n t o f th e R eq u irem en ts f o r th e D egree M a ste r o f A r ts by W a lte r Lew is H a ll A ugust 1938 UMI Number; EP67126 All rights reserved INFORMATION TO ALL USERS The quality, of this reproduction is dependent upon the quality of the copy submitted. In the unlikely event that the author did not send a complete manuscript and there are missing pages, these will be noted. Also, if material had to be removed, a note will indicate the deletion. iDisssartatfen R ib ltsW n g UMI EP67126 Published by ProQuest LLC (2014). Copyright in the Dissertation held by the Author. Microform Edition © ProQuest LLC. All rights reserved. This work is protected against unauthorized copying under Title 17, United States Code ProQuest LLC. 789 East Eisenhower Parkway P.O. Box 1346 Ann Arbor, Ml 48106 - 1346 This thesis, written by ....................MLTKR..MIS..HAI^^.................................. under the direction of h.l& Faculty Committee, and a p p ro ved by all its m em bers, has been presented to and accepted by the Council on Graduate Study and Research in partial fulfill ment of the requirements for the degree of MASTER OF ARTS Secretary 1 9 3 8 Date. Dean Faculty Committee Chairman A C K N O Y ilLED G EM EM PS The T f r ite r i s in d e b te d t o D r. C. E. T h ien es f o r h is in v a lu a b le a id a s ch airm an o f h i s co m m ittee; t o D o cto rs P a u l G reeley and F . M. B a ld w in .fo r t h e i r numerous s u g g e s tio n s and a id s ; to D r. Hermann S c h o tt f o r h i s k in d n e s s w h ile a s s i s t i n g him in t h i s w ork. The v jr i te r a ls o w ish e s to acknow ledge th e c o u r te s ie s ex ten d ed by D r. E . M. H a ll o f th e P a th o lo g y D epartm ent and e s p e c ia ll y t o Mr. Jo se p h M ard iro s o f t h a t d e p artm en t f o r h i s te c h n i c a l a s s i s t a n c e and to M iss G race Mas c h e r f o r h e r p a tie n c e and a id i n th e p r e p a r a t io n o f th e m a n u s c rip t. TABLE OP CO NTENTS , CHAPTER PAGE I . INTRODUCTION........................................' ................................................... 1 . I I . PE V im OF LITERA TU RE.......................................................................... 3 I I I . MATERIALS AND METHODS.......................................................................... 7 IV . EXPERIMENT.................................................................................................. 15 V. DISCUSSION OF F IN D IN G S ................................................. 30 V I. SU M M A R Y . .................................................................................................. 38 BIBLIOGRAPHY......................................................................................... 40 CHAPTER I imRODUCTION The r o l e o f g ly c o g e n i n c e l l u l a r p h y sio lo g y re m a in s to d a y a r a t h e r d e b a ta b le p ro b lem . I t i s g e n e r a lly acced ed t h a t th e p re s e n c e o f g ly c o g e n w ith in a c e l l i s accom panied by v i t a l a c t i v i t y and r a p id grow th e s p e c i a l l y d u rin g m u l t i p l i c a t i o n o f c e l l s . B ern ard (1855) d is c o v e re d g ly c o g e n i n th e l i v e r and b e lie v e d t h a t th e l a r g e amount found th e r e w as due t o th e em bryonic s t a t e o f t h a t o rg a n . P f liig e r (1905) and G lin k a (1911) w orking on em bryonic t i s s u e have shown g ly c o g e n a s a d e f i n i t e c o n s ti tu e n t o f em bryonic t i s s u e . As th e o rg an ism a g e s and th e t i s s u e p r o l i f e r a t i o n s become l e s s a c t i v e , th e q u a n tity o f g ly c o g e n d im in is h e s . Not much work h a s b e en done on g ly c o g en d e p o s its i n n e o p la s tic t i s s u e i n co m p ariso n t o th e v a s t amount o f l i t e r a t u r e t o be found u n d er tu m o r h e a d in g s . B ra u lt (1 8 9 8 ), one o f th e e a r l i e s t w o rk ers i n th e g ly c o g en o f m a lig n a n c ie s , found su ch d e p o s its i n a l l m a lig n a n t tum ors and i n a r e a s o f g r e a t p r o l i f e r a t i o n o f c e l l s . B est (1 9 0 5 ), on th e c o n tr a r y , found g ly c o g e n more p ro m in en t i n t h e slow grow ing tum ors and hence d oes n o t a g re e t h a t th e p re s e n c e o f g ly c o g e n , i n la r g e am ounts, i s c o r r e la te d w ith e x c e s s iv e c e l l a c t i v i t y . L u b a rsc h (1907) found g ly c o g e n i n 29 p e rc e n t o f a l l tum ors exam ined and o b serv ed t h a t th e sarco m ata c o n ta in e d more th a n th e c a rc in o m a ta . G ie rk e (1905) R ic k e r (1905) and F ic h e ra (1904) w ere among th e few o th e r s who worked a lo n g th e s e l i n e s , i n th e e a r ly p e r io d . The l a t e r w o rk e rs, d is c u s s e d i n th e re v ie w o f l i t e r a t u r e , b r in g th e work up t o d a te . E x p erim en ts w ith th e W alker Mamoiaiy R at C an cer have b e en done by McEuen and Thomson (1 9 3 3 -3 4 ) i n r e l a t i o n t o hypophysectom y and fo rc e d fe e d in g ; by Sam uels, B a ll and Sim pson (1932 and 1936) o n th e e f f e c t o f hypophysectom y on t h i s tum or and upon th e h o s t r a t . E a rle (1935) made a s tu d y o f th e s t r u c t u r e o f th e W alker R at Mammary C an c e r. S c h o tt, Sam uels and B a ll (1937) d id th e f i r s t work i n re g a rd t o g ly c o g e n d i s t r i b u t i o n i n tu m o r-b e a rin g r a t s . T h e ir r a t s w ere hyppphysectom ized and W alker R at C ancer No. 256 was u s e d . I t was t h i s ex p erim en t w hich su g g e ste d th e im p o rtan c e o f c o r r e l a t i n g th e ch em ical a n a ly s e s w ith th e h i s t o l o g i c a l d e m o n s tra tio n o f g ly c o g e n i n a n ad en o carcin o m a. The p r e s e n t r e p o r t , t h e r e f o r e i s a n a tte m p t t o make such c o r r e l a t i o n . The o n ly s tu d y o f t h i s s o r t found i n th e l i t e r a t u r e i s t h a t o f B orghi (1928) who in v e s t ig a te d th e g ly c o g e n o f a u t e r i n e adenocarcinom a by h i s t o l o g i c a l m eans. CHAPTER I I REVIEW OF LITERATURE The p re s e n c e o f g ly c o g e n i n m a lig n a n t tum ors was d e m o n strated by J . M u lle r by ch em ical means a s e a r ly a s 1838. L e b e rt and W alsh (1839) made f u r t h e r ch em ical, a n a ly s is o f c an c e ro u s g ro w th s, f in d in g a n abundance o f glycogen* B ra u lt (1845) showed t h a t th e r a p i d i t y o f grow th o f m a lig n a n t tu m ors was p r o p o r tio n a l t o th e amount o f g ly c o g e n th e y c o n ta in e d . De B âcher (1892) found t h a t s u b je c tin g a group o f g u in e a p ig s t o i n j e c t i o n s o f p u re y e a s t cau sed them t o become e x c e p tio n a lly f a t w ith accom panying s t e r i l i t y . He co n ce iv e d t h a t t h i s was due to a d im in ish e d p ro d u c tio n o f g ly c o g en i n th e u te r u s e t c . , co n seq u en t on y e a s t in j e c t i o n s . W ithout e x p e rim e n ta l p ro o f De B âcher co n clu d ed t h a t b la s to m y c e te s in tro d u c e d in t o a body, by u s in g up i t s g ly c o g en f o r th e m s e lv e s , m ight check p a th o lo g ic a l grow th by s ta r v i n g th e young c a n c e r c e l l s . G u llan d (1904) claim ed t h a t th e b lood o f c a n c e r p a t i e n t s i n t h e e a r ly s ta g e s o f th e d is e a s e i s f r e e from g ly c o g e n b u t a s th e m alady a d v a n c e s, * ’t h i s s u b sta n c e abounds i n it.** Lus c h e r (1907) h a s shown t h a t p a t i e n t s a r e so d e b i l i t a t e d from t h i s d is e a s e t h a t " th e g ly c o g en s and c a rb o h y d ra te s " a r e used up t o a n e x te n t w hich r e n d e rs c e l l p r o l i f e r a t i o n im p o s s ib le , c a u sin g a seem ing r e t r o g r e s s i o n o f th e tu m o r. L u b a rsc h (1907) found th e fo llo w in g c o n d itio n s f o r th e a p p e a r a n ce o f g ly c o g en i n tu m o rs: (1 ) O rig in from em bryonal t i s s u e s d u rin g u t e r i n e d ev elo p m en t. (2) O r ig in from c e l l s w hich n o rm a lly c o n ta in g ly c o g e n . (3 ) The a b se n c e o f m ucous and c o l l o i d a l tr a n s f o r m a tio n w hereby g ly c o g en and f a t d e p o s its o f te n a p p e a r to g e th e r . (4 ) The p re s e n c e o f numerous f i n e b lo o d v e s s e ls i n tum or stro m a and in n e r r e l a t i o n s h i p b etw een th e s e and tum or parenchym a, w hereby c i r c u l a t o r y d i s tu rb a n c e s a r e b ro u g h t a b o u t. R o llo (1913) found t h e r e was g ly c o g en i n th e m a lig n a n t, b u t n o t i n th e b e n ig n tu m o r. He th o u g h t t h i s t o be o f. d i f f e r e h t i a l d in g h o s tic 'a n d p r o g n o s tic v a lu e . ' B ern h ard (1928) found t h a t th e g ly c o g en c o n te n t o f m a lig n a n t tu m o rs was t e n tim e s g r e a t e r th a n i n u t e r i n e nyom ata. However mammaiy carcin o m a was e x d e p tio n a l i n h a v in g a low g ly co g en c o n te n t, p resu m ab ly due to i t s h ig h c o n n e c tiv e t i s s u e s t r u c t u r e , i t b e in g a s c i r r h o u s ' tu m o r. R oussy and C ra c iu n (1928) w o rk in g on th e Je n s e n R at Sarcom a co n clu d ed t h a t th e g ly c o g en c o n te n t was " r a t h e r d i s c r e t e , l i t t l e abund a n t , d i s t i n c t l y l o c a l iz e d and c e r t a i n l y l i m i t e d ." T h e ir work showed them t h a t th e g ly c o g en m a n if e s ts i t s e l f i n th e c e l l s o f th e tum or a s w e ll a s th o s e o f th e stro m a and t h a t th e s e c e l l s a r e s m a ll i n number i n p r o p o r tio n t o th e t o t a l m ass o f th e tu m o r. They found t h a t i n th o s e a n im a ls in o c u la te d w ith a s in g le g r a f t th e g ly c o g e n c o n te n t i s d i s t i n c t l y lo c a l i z e d a t th e l e v e l o f th e t h i r d zone (th e y found t h a t th e J e n s e n Sarcom a c o n ta in s f i v e z o n e s ) . They a l s o found t h a t th e g ly c o g e n b e a r in g - in g c e l l s w ere o f te n lo c a te d n e a r a fo c u s o f n e c r o s is and t h a t th e p a r t i c l e s found v a r ie d from f i n e re d g ra n u le s t o "some r a t h e r la r g e d r o p l e t s " . F u r th e r c o n c lu s io n s by th e s e men were t h a t th e g ly c o g e n ev o lv ed i n norm al c e l l s d u rin g grow th f o r a b r i e f p e rio d and t h a t i n p a th o lo g ic a l t i s s u e g ly c o g en a p p e a re d d u rin g d e g e n e ra tiv e c h a n g e s. Lund (1932) o b ta in e d g ly c o g en by p e p tic d i g e s t io n from l i v e r , embryo and tu m o r (sarco m a) t i s s u e and showed t h a t e a c h k in d o f g ly c o g en had a d i f f e r e n t r e f r a c t i o n . B abes and L a z a re s c u -P a n tz u (1930) i n a s tu d y o f a tum or o f th e c e r v ix show ed, i n agreem ent w ith L u b arsch (1907) t h a t th e amount o f g ly c o g e n had l i t t l e r e l a t i o n to th e p ro to p la s m ic d i f f e r e n t i a t i o n o f th e c e l l s a s claim ed by Lahm; a ls o t h a t th e g ly c o g en i s d i s t r i b u t e d a s th e c y to p la sm o f t h e c e l l . They a ls o assum ed t h a t th e p re s e n c e o f g ly c o g e n was a s s o c ia te d w ith th e a c t i v i t y o f th e c e l l . P e tro w a (1932) s tu d y in g myomatous t i s s u e o f th e womb d is c e rn e d t h a t th e g ly c o g en i s s to r e d i n th e m u scle c e l l s and a ls o i n th e c o n n e c tiv e t i s s u e and th e w a lls o f b lo o d v e s s e l s . He say s t h e r e i s a n e s p e c ia l abun d ance i n th e c o n n e c tiv e t i s s u e . He found g ly c o g en i n th e form o f sm all g ra in e d m asses a s w e ll a s i n th e sh ap e o f la r g e g r a in s and clum ps. McEuen and Thompson (1933) found t h a t hypophysectom y r e t a r d s t h e g ro w th o f tu m o rs i n r a t s , and t h a t m é ta s ta s é s a r e l e s s fr e q u e n t pro b a b ly due t o r a r e n e s s o f l a r g e tu m o rs. Body w eig h t i s a f f e c t e d . The same w o rk ers i n 1934 d e te rm in e d t h a t th e W alker R at tum or grow s a t same a v e ra g e r a t e i n r a t s o f a l l a g es (2 2 -9 0 0 d a y s ) . As th e r a t a g e s th e in c re a s e d gro w th o f th e tum or i s a t th e ex p ense o f th e e x i s t i n g so m atic t i s s u e s . S c h o tt, Sam uels and B a ll (1937) found t h a t th e slo w -g ro w in g tu m o rs o f t h e h y p o physectom ized r a t s c o n ta in e d l a r g e r am ounts o f g ly c o g en th a n th e tu m o rs o f th e i n t a c t a n im a ls . The g ly c o g e n c o n te n t o f th e l i v e r 6 and m uscle o f th e two g ro u p s showed l i t t l e d i f f e r e n c e . T hese d e te rm in a t i o n s w ere c h e m ic a l. The q u e s tio n s o f w h e th er th e g ly c o g en i n th e tum ors i s a s to r a g e phenomenon; why th e g ly c o g en v a lu e s w ere h ig h e r i n th e hypophysectom ized a n im a ls , and th e r e l a t i o n t h i s g ly c o g en had t o tum or m etab o lism a r o s e . T h ere i s a ls o th e q u e s tio n o f th e l o c a t i o n o f th e g ly c o g e n i n th e tum or and th e r e l a t i o n o f th e amount o f g ly c o g e n t o s i z e o f tu m o r. I n t h i s s tu d y a n a tte m p t i s made t o s o lv e some o f th e s e q u es t i o n s . CHAPTER I I I MATERIAL AND METHODS MATERIALS; The C arcinom as u sed i n t h i s r e s e a r c h w ere o f th e W alker R at Mammary C arcinom a No. 256, and w ere in o c u la te d i n t o some 300 m ale a lb in o r a t s by D r. H. A. B a ll o f th e San D iego County H o s p i ta l . The a v e ra g e age o f t h e r a t s was b etw een 1 2 -14 w eek s. A f te r in o c u la tio n , c e r t a i n o f t h e r a t s (a b o u t tw o - th ir d s ) w ere hypophysectom ized by D r. B a l l . A ll w ere in o c u la te d w ith th e tum or and w ere sh ip p e d t o u s h e re a t th e U n iv e r s ity o f S o u th e rn C a l i f o r n i a . Owing t o p o s t- o p e r a tiv e sh o ck , pneum onia and s ic k n e s s , a number o f th e a n im a ls w ere l o s t . T h is was e s p e c ia ll y t r u e o f h ypophysectom ized r a t s . METHODS; Upon r e c e i p t h e r e , th e a n im a ls w ere c a r e f u l l y w eig h ed , t h e i r tu m o rs m e asu re d . They w ere th e n s e g re g a te d a s t o hypophysectom ized tum or b e a r in g r a t s and norm al ( c o n tr o l) tum or b e a rin g r a t s . A ll a n im a ls w ere sto m a ch -fed tw ic e d a ily w ith a fo o d m ix tu re c o n ta in in g a h ig h s u g a r c o n te n t G lucose 100 p a r t s . Y e a s t..................... 15 p a r t s . P o W ered M ilk .................... . . . 1 0 0 p a r t s . W ater......................; . . . . . . . . . 3 0 0 p a r t s . Each r a t was w eighed and i t s tum or was m easured by c a l l i p e r s e v e ry d ay . The d a ta f o r each r a t was re c o rd e d on a s e p a r a te c a r d . When th e tum or re a c h e d a n ap p ro x im ate s i z e . o f f o u r t o f i v e 8 gram s i t was removed s u r g i c a l l y . Sodium am y tal was u sed a s a n a n e s t h e t i c b ecau se o f i t s s l i g h t e f f e c t on g ly c o g e n . I f th e o p e r a tio n was t o be on a hypophysectom ized an im al th e a d r e n a ls , gonads and head w ere r e moved p o s t- o p e r a t i v e l y . I n t h e c ase o f c o n tr o ls th e o p e r a tiv e i n c i s i o n was c lo se d and re c o v e ry o f th e an im al was a llo w e d . The e x tir p a te d tum or was c a r e f u l ly s e c tio n e d and a s e c t i o n a s n e a r ly th ro u g h th e m id d le a s p o s s ib le was re s e rv e d f o r h i s t o l o g i c a l p u r p o s e s . The re m a in in g m ass was used f o r q u a n t i t a t i v e e s tim a tio n o f th e g ly c o g e n p r e s e n t . B oth sam p les w ere w eighed t o d e te rm in e th e t o t a l w eig h t o f th e tu m o r. The p r e p a r a t io n o f th e t i s s u e f o r h i s t o l o g i c a l stu d y p ro ceed ed a lo n g th e l i n e s p r e s c r ib e d by B est (1903) f o r h i s carm ine m ethod: (1 ) Im m ed iately upon rem oval o f t i s s u e from th e an im al i t was p la c e d i n a b s o lu te a lc o h o l, and l e f t f o r 1 2 -1 4 h o u rs , d ep en d in g on s i z e o f sam p le. Two changes o f a lc o h o l w ere u sed d u rin g t h i s p e r io d . (2 ) The t i s s u e was imm ersed i n l / 2 x y lo l and l / 2 a b s o lu te a lc o h o l f o r 1 -2 h o u rs . T h is s te p i s n o t r e q u ir e d and was om m itted i n l a t e r w ork. (3 ) C le a rin g was done i n x y l o l . I f th e sam ple was sm a ll t h i s u s u a lly r e q u ir e d two h o u rs . L a rg e r sam ples to o k lo n g e r p e r io d s . Owing to th e d e n se n e ss o f c an c e ro u s t i s s u e , i t was n e c e s sa ry t o change th e s o l u t i o n a t l e a s t tw ic e . (4 ) I n f i l t r a t i n g i n p a r a f f i n (tw o ch an g es) r e q u ir e d 4 -6 h o u rs a t a te m p e ra tu re o f . a b o u t 55-60 d e g re e s . (5 ) T is s u e was p la c e d i n em bedding p a r a f f i n f o r a b o u t two h o u rs (5 8 -6 2 d e g r e e s ) . (6 ) A b lo c k was made and p la c e d i n th e ic e b o x o v e r n ig h t. (7 ) S e c tio n s o f 1 0 -12 mi e ra th ic k n e s s w ere c u t . (8 ) S e c tio n s w ere f l o a t e d on s l i d e s i n warm 90 p e r c e n t a lc o h o l a s g ly c o g en i s v e ry s o lu b le i n w a te r . (9 ) The s e c tio n w as f ix e d on th e s l i d e w ith egg album en and d r ie d f o r a t l e a s t f i v e d ay s i n a i r . Oven d ry in g te n d s t o d is s o lv e o u t g ly c o g e n . (10) S lid e s w ere s ta in e d i n f l a t c o p lin j a r s i n th e fo llo w in g m anner: (a ) P a r a f f i n was removed from th e s e c tio n s by p la c in g i n x y lo l f o r 1 -3 m in u te s . (b ) ]Qylol was rem oved from th e s e c tio n s by p la c in g them i n a b s o lu te a lc o h o l 1 -2 m in u te s . (c ) S lid e s w ere im m ersed i n 1 p e rc e n t c e l l o i d i n o v e r n ig h t. (d ) The s l i d e s w ere rem oved, d r a in e d , and allo w ed t o p a r t i a l l y d ry . ( I f th e y become to o d ry th e c e l l o i d i n te n d s t o f la k e and a f a l s e g ly c o g en a p p e a ra n c e i s g iv e n ) . (e ) S e c tio n s w ere d e e p ly s ta in e d i n H a r r i s ’ s h e m a to x y lin (a b o u t 5 m in u te s ) . 10 ( f ) D e s ta in in g was i n a c id a lc o h o l (1 -2 m in u te s ) , (g ) Washed i n ta p w a te r , (T h is was o m itte d i n l a t e r w ork a s th e ammonia i n th e carm ine s e r v e s th e same p u rp o s e , (h ) S ta in e d i n B e s t’ s carm ine ( d i l u t e d ) 2 -5 h o u rs , (L o n g er le n g th o f tim e i s p r e f e r r e d i n t h i s w o rk ), ( i ) D i f f e r e n t i a t e d i n B e s t’ s d i f f e r e n t i a t i o n s o lu tio n (tw o c h a n g e s ). The tim e elem en t h e re w as v a r i a b l e . K r a jia n (1936) s a y s t o l e t s l i d e s rem ain u n t i l re d c o lo r d is a p p e a r s . However t h i s seems a r a t h e r lo n g p e r io d . 1 -2 m in u te s i s recommend e d . ( j ) Washed i n 90 p e r c e n t a lc o h o l. (k ) D ehydrated i n a b s o lu te a lc o h o l (2 m in u te s ) . (1 ) C le a re d i n x y lo l o r x y lo l- a c e to n e . (P u re x y lo l p r e f e r r e d ) . (m) M ounted i n gum dam ar. S li d e s p la c e d on t r a y s , c o v e re d , and d r ie d f o r two w eek s. (Do n o t u se < d ry in g o v e n ). I n c a s e s w here a number o f tu m o rs w ere t o be ru n th ro u g h sim u l ta n e o u s ly th e p ro c e d u re was th e same e x c e p t t h a t th e tim e elem en t in s te p s 2 , 3 , 4 , 5, and 6 was d o u b le d . Gauze b ag s w ere n e c e s s a ry t o keep tu m o rs s e p a r a te . An a l t e r n a t e te c h n iq u e u sed i n t h i s p ro b lem was t h a t o f K in o s ita and D aikoku (1 9 3 6 ). The fo llo w in g p ro c e d u re i s t h e i r s : 11 (1 ) Im m ed iately upon rem oving tum or from a n im a l, p la c e i n f i x a t i v e c o n s is tin g o f 50 p e r c e n t a lc o h o l s a t u r a t e d w ith p o ta ssiu m c h lo r id e f o r from 9-14 h o u rs . (2 ) Remove to 70 p e rc e n t a lc o h o l. 1 -2 h o u rs . (5 ) Remove to 80 p e rc e n t a lc o h o l. l / 2 h o u r. (4 ) Remove t o 90 p e rc e n t a lc o h o l . l / 2 h o u r. (5 ) Remove to 95 p e rc e n t a lc o h o l . 1 /2 h o u r. (6 ) Remove to a b s o lu te a lc o h o l. 1 /2 h o u r. (Two c h a n g e s ). (7 ) P la c e i n x y lo l u n t i l c l e a r . (8 ) P la c e i n i n f i l t r a t i n g p a r a f f i n s . The re m a in d e r o f p ro c e d u re i s same a s i n B e s t’ s m ethod. The Langham’ s I o d in e S ta in f o r d e te rm in in g g ly c o g en was a ls o u s e d . T h is m ethod c o n s is ts o f th e fo llo w in g ; (1 ) D is s o lv e p a r a f f i n from s e c tio n i n x y l o l . (2 ) Remove t o a b s o lu te a lc o h o l. (3 ) S t a i n i n L u g o l’ s s o l u t i o n . 5-10 m in u te s . (4 ) D eh y d rate i n 1 p a r t t i n c t u r e o f io d in e t o 4 p a r t s a b s o lu te a lc o h o l. 5 s e c o n d s. (5 ) C le a r and d i f f e r e n t i a t e i n o i l origanum ( c r e t i c ) . (6 ) Mount i n gum dam ar. M o d ific a tio n s o f t h i s m ethod u se d in t h i s w ork: (1 ) A f te r p a r a f f i n i n (1 ) was d is s o lv e d o f f , th e s l i d e s w ere p la c e d i n 1 -2 p e r c e n t c e l l o i d i n 1 2 -14 h o u rs . (O v e rn ig h t p r e f e r r e d ) . (2 ) I n (3), m ix L u g o l’ s s o l u t i o n w ith p u re g ly c e r in e . 5 0 -5 0 . 12 SOLUTIONS: B e s t’ s G a m ic ë S t a i n f o r G lycogen; ' Carm ine ...................................... . . . 2 gram s P o ta ssiu m C a r b o n a t e .1 gram P o ta ssiu m C h lo rid e ....................... .5 grams D i s t i l l e d w a t e r 60 gram s Warm above m ix tu re f o r a few m in u te s , c o o l and add 20 c c . o f c o n c e n tra te d Ammonium l^ d r o x id e . Do n o t b o i l . Keep i n d a rk b o t t l e and c o o l p la c e . F i l t e r b e fo re u s in ^ . D ilu te w ith : B e s t’ s .2 6 c. C o n c e n tra te d Am. h y d ro x id e . . . . 3 c c . M ethyl a lc o h o l ...........................3 c c . B e s t’ s D i f f e r e n t i a t i o n : S o lu tio n : A b so lu te a l c o h o l ............................. 40 c c . M ethyl a l c o h o l ................................ .2 0 c c . D i s t i l l e d w a te r 50 c c . C e l lo id in S o lu tio n : P a r lo d io n (M a llin c k r o d t) . . . . . 1 gram . A b so lu te a lc o h o l . . . . . . . . . . . . . 5 0 c c . E th e r 50 c c . To d is s o lv e p a r lo d io n , s t i r b r i s k l y w ith heavy g la s s s t i r r i n g ro d and sh ak e c o n ta in e r o f t e n . T h is p ro c e d u re u s u a lly ta k e s a l l d ay . K in o s ita and D aikoku’ s S o lu tio n s ; (1 ) F i f t y p e rc e n t a lc o h o l s a tu r a te d w ith p o ta ssiu m c h lo r id e a s a f i x a t i v e s o lu t i o n . (2 ) F our p e r c e n t fo r m a lin s a tu r a te d w ith p o ta ssiu m c h lo r id e a s a f i x a t i v e s o lu t i o n . 13 L u g o l’ s S o lu tio n : I o d in e ( s o l i d c r y s t a l s ) 1 gram P o ta ssiu m io d id e . . . . . . . . . 2 gram s D i s t i l l e d w a te r .100 c c . DISCUSSION: Owing t o a few d i f f i c u l t i e s e n c o u n te re d d u rin g th e ex p erim en t and t o c l a r i f y some p o i n t s , a s h o r t d is c u s s io n i s n e c e s s a ry . The f i r s t group o f r a t s r e c e iv e d , h av in g num bers from 1 to 74, w ere u n d o u b te d ly n o t t y p i c a l o f th e p i c t u r e we w ish t o i l l u s t r a t e i n t h i s r e p o r t . O p e ra tiv e sh o ck , pneum onia and im p ro p er fe e d in g caused th e d e a th o f ^rnany. T h is was e s p e c i a l l y t r u e among th e h y p o p h y secto m ized . The re m a in in g l i v i n g a n im a ls w ith th e e x c e p tio n o f f o u r w ere c o n tr o ls . Removal o f th e tum ors was s u c c e s s f u l b u t te c h n iq u e was im proved on th e l a t e r a n im a ls . R e s u lts o b ta in e d from th e f i r s t group can n o t be*, t h e r e f o r e , a b s o lu te ly d e p e n d a b le . G roups I I , 1 1 1 , IV and V, how ever, w ere c a r r ie d th ro u g h a s a c c u r a te ly a s p o s s i b l e . The r a t s ch o sen from Group I w ere th o s e w h ich we th o u g h t would m ost p ro b a b ly y i e l d t r u e r e s u l t s . B ern h ard (1928) fo und t h a t mammary carcin o m a, o f a l l m a lig n a n t tu m o rs s tu d ie d , c o n ta in e d t h e l e a s t g ly c o g e n . T h is was b o rn e o u t i n t h i s r e s e a r c h an d due to th e d i f f i c u l t y i n o b ta in in g s u i t a b l e s l i d e s and to f u r t h e r m in im ize th e chance o f lo s in g w hat l i t t l e g ly c o g e n was p r e s e n t, s te p s ( f ) and (g ) i n th e B est M ethod w ere f i n a l l y e lim in a te d . The ammonia i n th e s t a i n i t s e l f w i l l com pensate f o r (g). I n s te p (h ) o f th e B e s t’ s m ethod th e s l i d e s w ere n o t allo w ed to re m a in u n t i l th e re d c o lo r d i s a p p e a re d a s recommended by K r a jia n (1 9 3 6 ). I t was found t h a t i f su ch was done th e d i f f e r e n t i a t i o n te n d e d to become d i f f u s e and t h e g ra n u le s o f 14 g ly c o g en w ere fa d e d . Due t o th e sm all.tam ouhts o f g ly c o g en found i n th e f i r s t and second g ro u p s o f t i s s u e by th e B est m ethod, th e te c h n iq u e o f K in o s ita and D aikoku (1936) was u s e d . T h is c o n s is te d o f s u b s t i t u t i n g f o r f i x a t i v e a s o lu t i o n o f 50 p e rc e n t a lc o h o l s a tu r a te d w ith p o ta ssiu m c h lo r id e , i n s te a d o f a b s o lu te a lc o h o l. When e v id en c e was o b ta in e d t h a t o n ly sm all am ounts o f g ly c o g e n w ere g o in g to be found th e fo rm er f i x a t i v e o f a b s o lu te a lc o h o l was re -e m p lo y e d . Langham’ s io d in e s t a i n p ro d u ces q u ic k e r r e s u l t s b u t th e s t a i n i s n o t p erm an en t, fa d in g o u t w ith in a p e rio d o f from t h r e e t o f o u r hours* N u c lea r d i f f e r e n t i a t i o n seem s t o be c l e a r e r when h e m ato x y lin i s u se d . CHAPTER IV EXPERIMENT DESCRIPTION OF TUMOR: U sing th e W alker R at Mammary C arcinom a No. 256 E a r l (1935) found t h a t th e o r i g i n a l p a r e n t tu m o r showed a t y p i c a l ad en o carcin o m ato u s s t r u c t u r e , r e t a i n i n g a g la n d u la r a r c h i t e c t u r e . The c e l l s o f e p ith e liu m c o n ta in e d la r g e rounded o r o v a l n u c le i w hich s ta in e d d e e p ly . The stroiM i o f th e tum or c o n ta in e d q u ite a n amount o f lo o s e f ib r o u s c o n n e c tiv e t i s s u e h a v in g many c e l l s . The m ost c h a r a c t e r i s t i c c e l l i s a r a t h e r e lo n g a te d s le n d e r f i b r o b l a s t - l i k e c e l l h a v in g n u c li o f v a ry in g s i z e . S li d e s o f t i s s u e from l a t e r g e n e r a tio n s showed a l o s s o f th e g r a n d u la r s t r u c t u r e b u t a r e t e n t i o n o f th e la r g e e p i t h e l i a l c e l l s a p p a re n t i n th e o r i g i n a l tu m o r. The stro m a o f th e su b se q u e n t tum ors v a r ie d from l a r g e am ounts t o s c a n ty , w id e ly d i s t r i b u t e d th r e a d s . C ro ss s e c tio n s o f su b cu ta n e o u s tu m o rs s i m i l a r i n ty p e t o th o s e u sed i n t h i s e x p e rim e n t, showed t h r e e z o n e s . The m ost p e r ip h e r a l one i s th e c a p s u le , a r e l a t i v e l y t h i n l a y e r su rro u n d in g th e tu m o r. Below t h i s i s th e r in d a r e a composed o f h e a lth y grow ing c e l l s . T h is zone i s in d e te rm in a te i n th ic k n e s s v a ry in g w ith d i f f e r e n t tu m o rs. The c e n t r a l zone E a r le found t o be alw ays n e c r o tic b u t g e n e r a lly q u ite s o l i d i n s t r u c t u r e . The c a p s u le was made up o f c o lla g e n f i b r e s and alm o st e n t i r e l y la c k e d r e t i c u l i n . Our e x p erim en t show ed, i n many c a s e s , a d e p o s it io n o f f a t i n th e c a p s u le a r e a s b u t maximow (1937) showed t h a t ro d e n ts have h ig h ly d ev elo p ed d e p o s its o f 16 brow n f a t t i s s u e i n th e s c a p u la r a r e a . Many o f th e tu m o rs w ere ta k e n from n e a r t h i s a r e a so t h a t th e f a t found may have b e en due t o an i n f i l t r a t i o n o f th e tum or i t s e l f r a t h e r th a n to a d e p o s it io n . The r in d zone i s e x tre m e ly r e t i c u l a r and d e n se ly f ib r o u s and u s u a lly h as a w h ite a p p e a ra n c e . HISTOLOGY: (C h a rt No. I ) Amount : ++ R at No. 1 . - i^ p o p h y s e etern ize d : I Tumor Wt: 5 .4 8 grams S iz e : 1 8 .5 Chemi s t r y : .0774$ N e c ro tic c e n te r ; s i c k l e shaped due t o l i q u i d c e n te r ; heavy stro m a c o n ta in in g narrow e lo n g a te d c e l l s . E vid en ce o f g ly co g en i n f in e f la k y g ra n u le s i n and n e a r n e c r o s is . Amount ; ++ R at No. 30 - C o n tro l: I Tumor Wt: 5 .5 5 grams S iz e : 1 7 .4 C h em istry : .081$ T y p ic a l heavy stro m a s t r u c t u r e ; many medium s iz e e p i t h e l i a l c e l l s ; few la r g e e p i t h e l i a l c e l l s and num erous s p in d le shaped c o n n e c tiv e t i s s u e c e l l s . G lycogen f a i r l y w e ll d i s t r i b u t e d th ro u g h many s e c tio n s i n s m a ll d r o p l e t s . L o c a ted f o r th e m ost p a r t i n th e r in d a r e a , c lo s e t o n u c le i o f l a r g e e p i t h e l i a l c e l l s . S m all n e c r o tic a r e a s o b serv ed i n c e n te r o f s l i c e o r s e c t i o n . 17 Amount : +++ R a t No. 56 - C o n tro l; I Tumor Wt: 1 .0 4 gram s S iz e : . S m all e x tr a Chemi s t r y : None. S tr u c tu r e s i m i l a r t o No. 3 0 . O c c a sio n a l sm a ll g ra n u le o f g ly c o g e n , seem in g ly i n no s e t a r e a . Amount : + R at No. 37 - C o n tro l: I Tumor Wt: 1 1 .3 8 gram s S iz e : 2 2 .0 C h em istry : .041$ Strom a s c a r c e ; p re v a le n c e o f la r g e p o ly g o n a l-sh a p e d e p ith e liu m c e l l s ; clum ps o f s m a lle r e p i t h e l i a l c e l l s ; n e c r o tic a r e a b e g in n in g i n c e n te r ; g ly c o g en s c a r c e , seem t o be i n n u c le i o f c e l l s in R ind zone; few s in g le d r o p le ts o r g r a n u le s . Amount : + R at No. 38 - C o n tro l: I Tumor Wt: 3 .8 0 grams S iz e : 1 4 .2 C h em istry ; .04 9 $ T y p ic a l s t r u c t u r e ; no n e c r o s is ; g ly c o g en , none o b s e r v a b le . Amount ; R at No. 40 - C o n tro l: I Tumor Wt : 4 .9 9 gram s S iz e : 2 4 .8 C h em istry : .0 6 2 $ L iq u id n e c r o tic c e n t e r . Tumor seemed t o have grown i n c o n c e n tr ic r i n g s ; b a r o r r i n g o f la r g e e p i t h e l i a l c e l l s s e p a ra te d by r i n g o f connec t i v e t i s s u e ; v e ry v a s c u la r on p e r ip h e r a l la y e r ; l a r g e d e p o s it o f g ly c o g en fo u n d i n s l i d e (2), e s p e c ia ll y i n f i r s t and second s e c t i o n s . R est o f 18 s e c tio n s seem in g ly v o id o f g ly co g en ; o c c a s io n a l la r g e g ra n u le o b serv ed i n o u te r p a r t o f r in d a r e a w hich l a y e r was d i f f i c u l t t o p e r c e iv e . Amount : R at No. 46 - C o n tro l; I Tumor Wt: 4 .7 1 gram s S iz e : 1 8 .4 C h em istry : .042$ L lm m ellar s t r u c t u r e a s i n No. 4 0 , a lth o u g h n o t a s la r g e a c e n t r a l l i q u i d n e c r o tic a r e a ; many la r g e e p i t h e l i a l c e l l s w ith c o rre sp o n d in g la r g e n u c le i . G lycogen: f a i r mount a s a w h o le. I n second s e c tio n o f s l i d e No. 1 th e r e a p p ea re d q u ite a p o c k et o f la r g e g r a n u le s . R e st o f s e c tio n s show o c c a s io n a l g ra n u le s a lth o u g h due t o p e c u l i a r s t r u c t u r e o f tum or i t i s h a rd t o say i n j u s t what p a r t i c u l a r l a y e r th e y make t h e i r a p p e a ra n c e . I n two o r t h r e e p la c e s g ra n u le s seemed t o be in s id e o f n u c le i , o th e rw is e , re m a in d e r w ere ly in g c lo s e t o n u c le i o f th e la r g e e p i t h e l i a l c e l l s . S m a lle r a r e a o f n e c r o s is seemed e v id e n t th ro u g h o u t s e c tio n o f tu m o r. Amount: +++ R at No. 55 - Ifypophysectom ized: I Tumor Wt: 3 .6 8 grams S iz e : 1 9 .9 C h em istry : .083$ S ic k le -s h a p e d s e c tio n s due t o la r g e n e c r o tic c e n te r w hich con ta in e d l i q u i d . U sual ty p e o f c e l l s a s d e s c rib e d by E a r le (1 9 3 5 ), b u t n o t c o n ta in in g a s heavy a stro m a a s p re c e d in g tu m o rs. G lycogen d e p o s its h eav i e r th a n any tum or th u s f a r exam ined. G ra n u le s o b serv ed w ere w e ll d i s t r i b u t e d th ro u g h o u t s e c t i o n s . Ntmierous p o c k e ts o r g ro u p s o f g r a n u le s i n r in d a r e a . Some w ere s i t u a t e d around n e c r o tic l i n e n e a r e s t c e n te r o f tu m o r. S m all g ra n u le s w ere o b serv ed in s id e n u c l e i , b u t t h i s o n ly 19 o c c a s io n a ll y . Amount: +t++ R at No. 60 - H ypophysectom ized: I Tumor Wt: 3 .9 5 gram s S iz e : 1 9 .4 C h em istry : .041$ T h is tum or had a heavy stro m a o f c o n n e c tiv e t i s s u e . The con ta in e d g ly c o g en seemed m a in ly c o n fin e d to clum ps o r p o c k e ts i n r i n d a r e a , s i t u a t e d a s u s u a l n e a r th e n u c le i o f th e la r g e e p i t h e l i a l c e l l s . One o r two clum ps w ere o b serv ed n e a r n e c r o tic b o rd e r. Amount : ++ R at No. 65 - % pophy s e c t omi zed : I Tumor Wt: 4 .3 6 gram s S iz e : 1 8 :1 Chemi s t r y : .042$ S e c tio n s from t h i s tum or show o c c a s io n a l la r g e g ra n u le s o f g ly co g en . One s e c tio n showed num erous c r y s t a l s o f v a r io u s s i z e s i n th e r in d a r e a , b u t c lo s e ly a s s o c ia te d w ith a n e c r o tic p a tc h . A few n u c le i seemed to c o n ta in sm a ll g r a n u le s . T h is tum or had a heavy stro m a , v e ry v a s c u la r n e a r c a p s u la r l a y e r and c o n ta in e d num erous n e c r o tic p a tc h e s . Amount : ++ R at No. 67 - C o n tro l: I Tumor Wt: 2 .9 4 gram s S iz e : 1 6 .0 C h em istry : .02 3 $ S ic k le -s h a p e d s l i c e due t o la r g e n e c r o tic c e n te r c o n ta in in g l i q u i d ; s m a ll amount o f stro m a ; c e l l s o f m ixed la r g e and sm all e p ith e liu m , to g e th e r ; g ly c o g e n seem in g ly s c a t t e r e d i n la r g e g ra n u le s o v e r tum or a r e a , b u t s c a r c e ; g ra n u le s c lo s e t o n u c le i o f la r g e e p i t h e l i a l c e l l s . 20 Amount; + R at No. 68 - C o n tro l: I Tumor Wt: 1 6 .2 2 gram s S iz e ; 2 7 .2 C h em istry : .024$ L a rg e ly e p i t h e l i a l i n s t r u c t u r e ; had a stro m a o f many th r e a d s o f c o n n e c tiv e t i s s u e ; l i q u i d n e c r o s is d e c id e d ly o f f c e n te r ; r in d zone c o n se q u e n tly d i s t o r t e d , c o n ta in in g many n e c r o tic p a tc h e s ; No g ly c o g en g ra n u le s o b se rv e d . Amount : - E a t No. 70 - C o n tro l: I Tumor Wt: 2 .8 5 graBis S iz e : 1 8 .0 C h em istry : .077$ c o lla g a n o u s f i b r e s th ro u g h o u t, e s p e c ia ll y n e a r to p ( s k in s id e ) o f tu m o r. E x trem ely l a r g e e p i t h e l i a l c e l l s and a few sm a ll o n e s. Tumor l a r g e l y n e c r o tic g iv in g r i s e to b a d ly chopped up r in d a r e a . Few c r y s t a l s o f g ly c o g e n found i n th e s c a t t e r e d p a tc h e s o f r in d m a t e r i a l . Amount: ++ R at No. 73 - C o n tro l: I Tumor Wt: 4 .4 0 gram s S iz e : 1 6 .9 C h em istry : .056$ L a rg e ly composed o f e p i t h e l i a l s tr a n d s ; no o b s e rv a b le n e c r o s is ; few c o n n e c tiv e t i s s u e s tr a n d s ; r in d a r e a seem in g ly t o compose m ost o f tum or w ith e x c e p tio n o f c a p s u le . G lycogen was o b s e rv e d , b u t lim it e d to s i n g l e sm all g ra n u le i n w id e ly d i s t r i b u t e d s e c t o r s . Amount : + R at No. 74 - C o n tro l: I Tumor W t: 8 .7 8 gram s S iz e : 2 3 .8 C h em istry : .056$ 21 Tim or composed o f l a r g e e p i t h e l i a l c e l l s , f i n e stro m a th ro u g h o u t; num erous p a tc h e s o f n e c r o s i s . G lycogen g ra n u le s s c a rc e h u t some o b serv ed i n r in d a r e a , e s p e c ia ll y t h a t r in d su rro u n d in g c e r t a i n n e c ro t i c p a tc h e s . Amount : + R at No. A - C o n tro l: I I Tumor Wt; 6 .6 0 gram s S iz e : 1 7 .9 C h em istry : .0759% T h is tum or had a n ev en m ix tu re o f c o n n e c tiv e t i s s u e stro m a and e p i t h e l i a l s tr a n d s . M oderate amount o f medium s iz e d g ly c o g en g r a n u le s , c lo s e ly a s s o c ia te d w ith t h e n u c le i o f th e la r g e e p i t h e l i a l c e l l s i n th e r in d a r e a . O c c a sio n a l g ra n u le s found i n t i s s u e s u rro u n d in g n e c r o tic p a tc h e s o f w hich th e r e w ere many. Amount : ++ R at No. B - C o n tro l; I I Tumor Wt: 5 .0 2 gram s S iz e : 1 8 .3 C h em istry : .0 5 5 ^ I r r e g u l a r shaped, tu m o r, due t o l a r g e n e c r o tic a r e a c o n ta in in g l i q u i d and lo c a te d n e a r s k in th ro u g h w hich i t b ro k e . S im ila r i n s t r u c t u r e t o tum or ”A” . G lycogen co m parable to tum or **A^. Amount : ++ R at No. 80 - % -pophysectom ized: " I I Tumor Wt: 4 .2 3 gram s S iz e : 1 6 .9 C h em istry : .9 5 ^ M ainly e p i t h e l i a l c e l l s t r u c t u r e w ith few in te rw o v e n s tr a n d s o f c o n n e c tiv e t i s s u e . One o r two r a t h e r la r g e n e c r o tic p a tc h e s . W idespread e v id e n c e o f g ly c o g e n g ra n u le s o f a l l s i z e s , g e n e r a lly in te r s p e r s e d 22 th ro u g h o u t s e c t i o n s . Amount: ++++ R at No. 82 - i^ p o p h y s e ato m ized : I I Tuiiior Wt: 4 .7 9 gram s S iz e : 1 8 ,3 C hem istry : •114^ S m all tu m o r, l a r g e l y composed o f c o n n e c tiv e t i s s u e s tr a n d s . Many g ly c o g e n g ra n u le s d i s t r i b u t e d th ro u g h o u t s e c t i o n s . Few n e c r o tic p a tc h e s s t a r t i n g . Amount: ++++ R at No. 84 - H ypophyseato m ized : I I Tumor Wt: 4 .6 7 gram s S iz e : 1 8 .4 C h em istry : .082^ E x trem ely heavy c o n n e c tiv e t i s s u e s t r u c t u r e c o n s is tin g o f r i v e r - l i k e s tr a n d s o f t h i s m a te r ia l th ro u g h o u t s e c t i o n s . Many c o llo id is la n d s o b serv ed n e a r s k in s id e o f tu m o r. Many n e c r o tic p a tc h e s , v a ry in g i n s i z e ; s tr a n d s o f e p ith e liu m c o n s is te d , f o r th e m ost p a r t to be o f th e s m a lle r c e l l ty p e . G lycogen g ra n u le s o f medium s i z e , se e n i n g ro u p s i n v a r io u s p a r t s o f s e c t i o n s . Some g ra n u le s seemed t o a p p e a r i n th e con n e c tiv e t i s s u e s tr a n d s , and some seem in g ly e x t r a - c e l l u l a r . Amount : +++ R at No. 88 - ^y p o p h y secto m ized : I I Tumor Wt: 4 .0 6 gram s S iz e : 1 7 .9 C h em istry : .114$ S ic k le -s h a p e d s e c t i o n due t o la r g e l i q u i d n e c r o tic c e n te r ; heavy c o n n e c tiv e t i s s u e stro m a . U su al l a r g e e p i t h e l i a l c e l l s in te rm ix e d . S eem ingly heavy g ly c o g en g ra n u le d i s t r i b u t i o n , n o t p a rtic u la ris m c o n fin e d t o one l a y e r . 23 Amount: ++++ R at No. 95 - I^ypopliy s e c t om ized: I I I Tumor Wt: 1 .0 6 gram s S iz e : 1 8 ,9 C h em istry : . 0 0 ^ L arg e amount o f c o n n e c tiv e t i s s u e la c k in g i n q u a n tity o f th e u s u a l f i b r o b l a s t - l i k e c e l l s . Many la r g e e p i t h e l i a l c e l l s . G lycogen g ra n u le s sc a n ty and v e ry s m a ll. No d e f i n i t e e v id en c e o f n e c r o s is and r in d zone seems to occupy e n t i r e c e n t r a l a r e a . Amount : + R at No. 97 - C o n tro l: I I I .Tumor A Tumor Wt: 2 .2 3 gram s S iz e : 1 8 .2 C h em istry : .029$ T h is tum or was d a rk i n a p p e a ra n c e . F a i r amount o f c o n n e c tiv e t i s s u e stro m a p lu s l a r g e e p i t h e l i a l c e l l s . I r r e g u l a r shaped s e c tio n s due t o l a r g e l i q u i d n e c r o s is to w a rd s s k in s id e o f tum or ms-king la y e r s d i f f i c u l t t o p e r c e iv e . G lycogen c o n fin e d t o n u c le i o f e p i t h e l i a l c e l l s . G ra n u le s la r g e b u t n o t p r o f u s e . Amount: ++ R at No. 97 - C o n tro l: I I I Tumor B Tumor Wt; 3 .3 2 gram s S iz e ; 1 2 .6 C h em istry : .084$ S can ty stro m a ; l a r g e l y composed o f e p i t h e l i a l s tr a n d s . Sm all n e c r o tic c e n te r . G lycogen made up o f medium s iz e d g ra n u le s and s c a tte r e d g e n e r a lly th ro u g h o u t s e c t i o n s . S eem ingly to be i n g r e a t e r number th a n i n s i s t e r tum or No, 97, A. Amount : +++ 24 R at No, 98 - C o n tro l: I I I Tuiaor Wt: 5 .1 0 grams S iz e : 1 8 .6 C h em istry : .019$ F a i r amount o f stro m a . Numerous la r g e e p i t h e l i a l c e l l s and o c c a s io n a l p a tc h e s o f sm a ll o n e s. R ind a r e a i r r e g u l a r and w here i t i s sp re a d o u t, n e a r b ase o f s e c tio n , t h e r e a p p e a r q u ite a few s in g le medium s i z e g ly c o g e n c r y s t a l s . L arg e n e c r o tic a r e a b e g in n in g o f f c e n te r o f tum or n e a r to p o r s k in s id e . Amount : ++ R at No. 105 - H ypophysect om ized: IV Tumor Wt: 8 .8 0 gram s S iz e : 1 9 .5 C h em istry : .022$ U sual d i s t r i b u t i o n o f lo o s e f ib r o u s c o n n e c tiv e t i s s u e and la r g e e p i t h e l i a l c e l l s . Few sm a ll e p i t h e l i a l c e l l s . G lycogen lim ite d t o oc c a s io n a l s in g le sm a ll g ra n u le s lo c a te d n e a r n u c le i o f th e la r g e e p i t h e l i a l c e lls * T h is tum or was d a rk i n a p p e a ra n c e . Amount : + R at N o.109 - Bfcrpophysectomized: IV Tumor Wt: 2 .5 3 gram s S iz e : 1 9 .4 C h em istry ; .012$ A d a rk a p p e a rin g tum or c o n ta in in g a p ro fu s e d i s t r i b u t i o n o f f ib r o u s c o n n e c tiv e t i s s u e . Many la r g e e p i t h e l i a l c e l l s in te m ix e d w ith sm a ll o nes i n te r s p e r s e d th ro u g h o u t. Numerous n e c r o tic p a tc h e s . G lycogen g r a n u le s r a r e and c o n fin e d t o n u c le i o f th e la r g e e p i t h e l i a l c e l l s . Amount : + R at N o.112 - C o n tro l: IV Tumor Wt: 4 .9 2 gram s S iz e : 1 1 .8 C h em istry .072$ 25 S iia ila r i n s t r u c t u r e t o No. 109 e x ce p t t h a t th e la r g e , e p ith e l i a l c e l l s w ere more c lo s e ly packed and tum or was n o t o f d a rk a p p e a ra n c e . C e n te r o f tum or had n e cro se d le a v in g a l i q u i d . G lycogen co u ld be com p a r a b le t o amount s e e n i n No. 109 e x c e p t t h a t g r a n u le s a p p e a re d , more o f te n , s i n g l e . Amount : + R at No. 247 - I^ p o p h y se cto m ize d : V Tumor Wt: 4 .4 4 gram s S iz e : 1 7 .2 Chemi s t l y : . 100$ L a rg e ly f ib r o u s c o n n e c tiv e t i s s u e s t r u c t u r e w ith u s u a l numerous la r g e e p i t h e l i a l c e l l s . B eg in n in g o f n e c r o s is i n c e n te r o f tum or b u t t h a t a r e a s t i l l f ir r a . F in e g r a in s o f g ly c o g en w e ll d i s t r i b u t e d o v e r r in d a r e a b u t none in g r e a t am ounts. Amount : ++ R at No. 253 - C o n tro l: V Tumor Wt: 4 .0 5 gram s S iz e : 1 7 .2 C h em istry : .056$ V ery lo o s e s t r u c t u r e a s i f w hole tum or was b re a k in g down. Many s tr a n d s o f f ib r o u s c o n n e c tiv e t i s s u e w ith clum ps o f la r g e e p i t h e l i a l c e l l s . G lycogen g ra n u le s e v id e n t i n .s p o t s b u t no d e f i n i t e a re a can be a s s ig n e d owing t o tum or s t r u c t u r e . Amount : R at No. 300 - % pophy s e c t om ized: V Tumor Wt: 5 .1 3 gram s S iz e : 1 8 .0 C h em istry : .08 9 $ S tr u c t u r e s im ila r t o No. 255 e x ce p t f o r a h e a v ie r p a c k in g o f th e e p i t h e l i a l c e l l s and a p p e a ra n c e o f many c o llo id d e p o s it s . G lycogen 25 g ra n u le s w ere r a t h e r la r g e and lo c a te d i n th e e p i t h e l i a l clum ps. One la r g e d e p o s it o b se rv e d n e a r c o llo id m a t e r i a l . Amount ; +++ R at Kb. 349 - J^ p o p h y secto m ized : V Tumor Wt: 4 .3 6 gram s S iz e : 1 7 .0 C h em istry : 0 .4 0 $ S tr u c t u r e more t y p i c a l . M a jo rity o f la r g e e p i t h e l i a l c e l l s w ith clum ps o f s m a lle r ty p e . N e c ro tic p a tc h e s a p p e a r i n d i f f e r e n t a r e a s th ro u g h p u t s e c t i o n s . G lycogen i n f i n e g r a i n s . W ell d i s t r i b u t e d i n r in d a r e a . Amount: +++ R at No. 374 - H ypophysectom ized: V Tumor Wt: 5 .4 3 gram s S iz e : 1 7 .4 C h em istry : .080$ T y p ic a l f ib r o u s stro m a , b u t w ith a seem ing predom inance o f th e s m a lle r e p i t h e l i a l c e l l s . L arg e l i q u e f a c t i o n o f c e n te r o f tum or due to n e c r o s i s . G lycogen com parable t o Kb. 349. + 1 o o o o o o o o I I 88 s f % c - 0 0 to s‘ >C O si' C D o o C D o C D C D C D + + + + 1 * « P - I - + + + + - 4 - 4 + + + + + 1 •r i E Q p P C D < H rP 1 — 1 0 5 r H + 3 P 1 g r a O C D « r i 6 0 5 P « r i r - 4 4i O A o « r i p m Ü O P P 03 03 o ( — 1 C D P 1 O p c d P i C d « r i P Pi 4i 0 3 p -P P Pi T d P C D Ü P C D« r i P O r H p • H P Ü fi P P i C D P i r-i f H r H p P Pi P % P c d fl ( d O P -P « r i C D p • H P P p P P P P Ü 1 A ri fp P i T d « + = 03 r H > p P 'P Pi * P P i C d P • H O r H p •P P P P « H C D« r i o A JQ P « r i P « r i P P • H « P Pi + a Q ) a Î S Î « r i p :i M Pi P < t - i Pi W C s 2 O O ' sji O C Q A S G ) O O +> O C O a) . ® Ü 0) A » H fy C d 4* < D O -P •a S 'S •H 0 ) O « « to 8 '3 •H o to t o m m o o + + 0 5 CO ( D * H X S -P O CD + 3 P > + 5 ^ I 'd c d (D C D f H A CD CO c o a O -H C Q K % ^ C' l O o o § s ■ a « c O ( D 5 % *r4 « Pi p P - p « r i C Q p S’ # H p P T d C Q o P p O P 03 P W P i T d P P p Q ) P o ri iJ rP P P -P Ü -p Ü * r 4 -P A P ■ P P ■P b O P « r i Ü P C D P P P h 4 C Q A C Q APQ o m ^ ^ a fl C D C D C D o o o n d 'T O -O •H * M « H E 5. c o < C 3 ^ e r r * «ri «ri « H h4 1 - 4 A •H g a A C D " S o C D C D pq o TO C D ë -p "S o C Q S I o «ri h4 (0 < P P H £ A < P P P T d T d o P O P p o P rP P P P P 'O P % W ) P P p P T d P « r i P p 1 P « P p « P * H P P -p o P Ü p Ü ■ S c P P K 7 p P P P P c r * a •ri P p X i p ( d Ü P « r i p h 4 Ü > •p C Q A I Q A 1 - 4 o I + + + + « * « + + + + + + * + + + + + + + + + + H - + + + + - ► + « * " + « K + + . . p P P P p P P P P P P P P p p P P 00 00 Î 5 Û bO H D H O HO HO HO HO HO H O H O HO HO H O H O 00 lO 00 O < T > r H 00 C Q to Hc C M Q C O o C M ID o C O 00 CH CS to C Q <n C M 00 ^ IS to O m lO 1 — 1 rH C O Hc sj* C Q C Q C M to C M 00 to to rH r H , . p , , , , , , • • • ' p o • p O o P P P P P P P O ' • p p p p o o o o P P P P P P P o P P p p C O M O C M 00 CJ) . r H O C M O < T > 00 C T > C Q 00 IS C M sM Hc 00 c n Cft 00 to o - 00 to C Q IS CO r H r H O j C M rH C M r H r H rH r H H cM 1 — 1 rH C M r H rH o to 'IS t o C O O to lO lO ^ lO o to lO to IS C O to to g P Q 'g N •H § « C D C Q o ^ Pi "ë P i o Î& I I I k O ÎIÎ + 1 + I ta w ta ta ta o o o ta ta o ta % % % % H œ r-4 0 oi c o rH 0 2 rH <T > r-4 a r4 0 0 0 0 0 C D S 3 Si O O % I T d - « d (d 0) c d < D P ( D p A P ( d ta C C S C D T d Td -p cî • H 0 ) - H ^ C Q p d g p i •H 1 i ( D P a -H ta c d C Q 4.4 • H I 1 -P td o C Q a 0 o C Q 0 C Q a - p • H 1 § + + + « * « 4- 4 4 4 4 + 4- 4- 4 4 4 4- + ta < — 1 w 1 — 1 a a 1 — ! « P * P r H A •ri H O A a a a 0 r H p a .a « r i P a a p P « r i si r H r H a 4 3 r H Td a a A a Td a r H •r i A a a a a «H a 0 « r i a a P A 0 •ri a a p 0 r H p P a a 1 — 1 P p a t > > b d « r i a a a a « r i a a a « p H a 4 3 a ta a si 4 3 0 a « a r H 4 3 4 3 ' p p H O 1 — ! r H 43 ■ a a * f 4 a a P « r i a a a P r H P •ri a a p C Ü 0 •ri A p • r-4 a a A A « r i •ri C Q a N P 4 S § a c Q « • 2 5 r H p 5 a A P i ta a g p a sa •r i 43 P p a a A 43 a T d a A 43 a •r i a p 0 A H O a P a P a a a a T d T d 0 a si 43 .a « ri 1 « H H O p 43 p a a a p > 43 a p c d tj* a a a a p a •ri « r i 0 r H A C Q A pj S 0 } o i ë H O "ë < D C D < P < p O ^ a a>£ •H a m H O o m p a C D P "ë a 0 ) r4 p a « r i a T d a a a P ü a p p a p A a p g C Q C Q ë -p + 3 a p ( Q ë " ë p p C Q ,S O +3 Td a * H & a T d " H s ( D K - q 1 ë * S ô " ë C D a p q p g E - t & C Q 0 . 7 « s t » S S) c r > A œ 4 • 4 4 4 4 4 ' 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4- + 4 4 p P P P P P P P p p H O H O H O H O H O H O H O H O H O H O £S tO C O C O 02 0 Q IQ 0 2 tO 0 lO 02 C O r H 00 10 03 « s t » H » tj» H » C O 10 c o I D - s j » - s t » P P P P P P P p P P P P P P P P P P p P P P 03 C O 03 02 tO tO 00 07 C D 00 00 C D C O 03 00 03 C J 3 A I S r — \ rH A A A A rH A A A A s 1 tn es c j i c r > p q I es C D 1 0 03 02 es 0 » C T > 0 0 A ^ A A A W T d a N « H g -P P a ta o è - p A "ë A O t& I l I I o ta + I +1 o td td td to t i ' J c o o o SI o o + + + + 4* + + + 4 4 4 S i 3 8 O Ü O " a a P a * 2 a A a a •ri a 4 > O a • H a A a P a a A A a a •ri p O a p A a O •ri a A a P a T d A P a A •ri a a A a C d A • H a a •ri S « a 6 td ë •p a A a a o A 1 — 1 P a a a A p p A A a a A a a a a C d * a a ü •ri O C i J C Q h d i C Q 4 4 4 4 4 4 4 4 4 P P P P H O H O H O H O æ \0 C Q t O D O C 5 O A C O A A a H T t » l O « S f i l O N • r i S o A P O p P P a o p P P ta M A t S 3 03 O O -tf p Hd A P a, C Q O * 00 I S O •p o A A A A a P 4 O t& H I I A O td O g D O O l O o 03 « s j » •sji IS 03 C O to to + 1 A 8 CHAPTER V DISCUSSION OF FINDINGS The d e p o s itio n o f g ly c o g en i n th e W alker R at Mammary C arcinom a seem s to be g r e a t e s t i n th e second o r r in d l a y e r . T h is a re a r e p r e s e n ts th e fo c u s o f h e a lth y c e l l s and th e m ost p r o l i f e r a t i n g . B ra u lt (1898) L u b arech (1907) and Babes and L a z a re s c u -P a n tz u (1950) a l l c laim ed t h a t th e g r e a t e s t amount o f g ly c o g e n was found w here th e c e l l s w ere m ost a c ti v e and p r o l i f e r a t i n g . B est (1903) c o n tr a r y t o o th e r w o rk e rs i n t h i s f i e l d claim ed t h a t he found g ly c o g e n m ore p ro m in en t i n th e slo w ly grow ing tu m o rs. (F ig u re 1 ) . T h is e x p erim en t gave s u p p o rt t o B est *s c la im b u t b e ca u se o f v a r i a t i o n s among th e tu m o rs, a la r g e number w i l l have to be s tu d ie d t o p ro v e th e p o i n t . The amount o f g ly c o g e n found was e x tre m e ly sm all i n co m parison t o g ro s s s i z e and w eig h t o f tu m o rs, s u p p o rtin g th e a s s e r t i o n s o f B ernhard (1928) t h a t i n mammary carcin o m a one f in d s th e l e a s t d e p o s its o f t h i s s u b s ta n c e . (F ig u re 2 ) . B ern h ard claim ed t h a t t h i s was p ro b a b ly due to mammary carcin o m a h av in g a heavy c o n n e c tiv e t i s s u e s t r u c t u r e . I t i s t r u e t h a t i n t h i s tum or lo o s e f ib r o u s c o n n e c tiv e t i s s u e i s abu n d an t b u t a s i l l u s t r a t e d by tum ors No. 82 and No. 55, th e g ly c o g e n c o n te n t was g r e a t e r th a n i n some, n o t e x h ib itin g so much s tro m a . • The c r y s t a l s o r g r a n u le s o f g ly c o g en te n d t o b e i n th e cy to p lasm o f th e l a r g e e p i t h e l i a l c e l l s , ly in g c lo s e t o th e n u c le u s . (F ig u re 3 ) . F ig u re 1. MicroT)hotograph o f Tumor Ko* 84 I^pophysectoraized showing glycogen d i s t r i b u t i o n . T his wes a slow growing tumor, (M a g n ific a tio n 440X). F ig u re 2, M icrophotograph of Timor No. 95 C o n tro l showing sca n ty d i s t r i b u t i o n of glycogen in com parison to s iz e and w e ig h t. T his tumor- weighed 4.06 grams and was 18.9 cc, ill s iz e , (M a g n ific atio n 440X}. ï ’ ig u re 5* M icrophotograpli o f Tumor No* 56 C o n tro l showing A la r g e g ran u le A SL iB 1 ],e r g ra n u le g ra n u le s o f glycogen i n v a rio u s lo c a t io n s , i s shown in th e strom a which i s very r a r e , may be observed c lo se to th e n u cleu s o f a la r g e e p i t h e l i a l c e l l . T his i s th e u s u a l p o s i t i o n o f th e g ra n u le s , h n o tijer, s t i l l sm a lle r g ra n u le , may be seen c lo se to th e n u cleu s of a n o th e r la rg e e p i t h e l i a l c e l l , alm ost in s id e of n u c leu s, (M a g n ific a tio n 440X)• 54 O c c a s io n a lly sm a ll g ra n u le s w ere found i n th e n u c le i and r a r e l y i n th e c o n n e c tiv e t i s s u e stro m a . R oussy and C ra c iu n (1928) i n t h e i r work on t h e J e n s e n Sarcom a found c a s e s w here th e n u c le i w ere lo a d ed w ith g ly c o g e n . They a l s o found m a n if e s ta tio n s o f g ly c o g e n m ore f r e q u e n tly i n th e stro m a t h a t t h i s ex p erim en t d id n o t b r in g out* However th e r e i s a n agreem ent betvfeen th e tw o r e s e a r c h e s a s t o th e amount- fo u n d and l o c a l i t y o f th e s e g r a n u le s . A lso th e y c la im t h a t g ly c o g en b e a rin g c e l l s w ere o f t e n lo c a te d n e a r a fo c u s o f n e c r o s is . T h is ex p erim en t showed t h a t g ly c o g e n g ra n u le s w ere p r e s e n t i n n e c r o tic a r e a s b u t th e s e a r e a s w ere u s u a lly i n th e r e g io n o f sm all n e c r o tic p a tc h e s b e g in n in g i n th e r in d a r e a and w ere n o t f r e q u e n t l y s e e n . A lth o u g h h i s t o l o g i c a l r e s u l t s c o r r e la te d w ith th e ch em ical a n a ly s e s , w ith th e e x c e p tio n o f Kb. 70 C o n tro l and No. 247 Hypophy s e c t om ized, t h e r e was a p p a r e n tly n o t a s much g ly c o g e n d e m o n stra b le by h i s t o l o g i c a l m eans a s was found by c h em ica l a s s a y . L iq u e f a c tio n o f th e c e n te r o f th e tum or a s th e r e s u l t o f n e c r o s is may e f f e c t th e amount o f g ly c o g e n found h i s t o l o g i c a l l y . E a r le (1935) c o n te n d s - th a t a lth o u g h th e c e n t r a l zone was alw ays fo und t o be n e c r o tic th e s t r u c t u r e was g e n e r a lly q u i t e s o l i d . F u r th e r ex p e rim e n t seems t o show t h a t t h i s i s n o t alw ay s th e c a s e . O ver o n e - h a lf th e tu m o rs o b ta in e d i n t h i s p r e s e n t work showed d e f i n i t e l i q u e f a c t i o n o f ;bhe tum or c e n te r due t o n e c r o s is , v' *. The ch em ical e s tim a tio n o f th e tum or g ly c o g e n showed t h a t th e r e was no m arked d if f e r e n c e i n am ount o f t h i s s u b s ta n c e betw een th e bypoplQrsec- to m iz e d and c o n t r o l s . T h e re was a te n d e n c y , how ever, on th e p a r t o f th e l^ p o p h y se cto m ize d t o have a s l i g h t amount g r e a t e r th a n th e c o n t r o l s . T h is 35 was b o rn e o u t, a l s o , i n th e h i s t o l o g i c a l s e c t i o n s . (F ig u re 4 ) . S c h o tt, Sam uels and B a ll (1937) found t h a t th e a v e ra g e g ly c o g en c o n te n t o f th e h y p o p ty sec to m iz e d r a n h ig h e r th a n d id t h a t o f th e c o n t r o l s . I n t h e i r work .the tum ors w ere not a s w e ll c o n tr o lle d a s t o s i z e a s was done i n th e p r e s e n t w ork. F u r th e r , th e s e w o rk ers w ere com paring th e d i s t r i b u t i o n g ly c o g e n i n th e tu m o rs w ith th e d i s t r i b u t i o n o f t h i s s u b sta n c e i n l i v e r and m u sc le . I n th e p re s e n t w ork s e p a r a te e s tim a tio n o f g ly c o g e n was made f o r th e o u ts id e l a y e r o f tum or and f o r th e in s id e o r m ost n e c r o tic l a y e r . T h is was n o t done i n th e p re v io u s e x p e rim e n t. C h em ically th e r e s u l t s o f d iv id in g th e tum or i n t o two p a r t s showed t h a t a s a n a v e ra g e th e o u ts id e p o r t i o n (w hich e v id e n tly c o n ta in e d much o f th e r in d l a y e r ) con^ t a i n e d th e m ost g ly c o g e n . O c c a s io n a lly th e n e c r o tic p o r ti o n was h ig h i n th e c o n te n t o f t h i s s u b s ta n c e . T h is ex p erim en t showed t h a t hypophysectom y d id r e t a r d , t o a c e r t a i n e x te n t , th e g row th o f th e tu m o rs and th o s e r a t s so o p e ra te d on w ere s u b je c t t o c o ld s , d i g e s t iv e d is tu r b a n c e s and w ere p ro n e t o a t t a c k s by m ite s . T h is a g re e s w ith t h e work o f McEuen and Thompson (1 9 3 3 ). C e r ta in o f th e tu m o rs ( F ig u re 55) showed a darfc a p p e a ra n c e o f th e t o t a l m a s s .' T h is was c o n tr a r y t o th e u s u a l l i g h t a p p e a ra n c e o f th e g ro s s tum or and th e u s u a l tr a n s l u s c e n t w h ite c o lo r on s e c tio n in g . T hese tu m o rs w ere n o t a s fir m i n c o n s is te n c y b u t c o n ta in e d th e u s u a l heavy s tro m a . The g ly c o g en c o n te n t seemed t o be lo w er th a n o th e r tu m o rs com parable i n s i z e and w e ig h t. O b se rv a tio n s seemed t o show t h a t th e s m a lle r tu m o rs c o n ta in e d th e g r e a t e s t amount o f g ly c o g e n on a co m p arativ e b a s i s . F ig u re 4 , Two m icrophotographe showing glycogen d i s t r i b u t i o n in two ty p e s of tu m o rs. The upper p ic tu r e shows d i s t r i b u t i o n of glycogen i n a tumor from a hypophysectom ized an im al. T his tumor was ra te d a t 4+ h i s t o l o g i c a l l y . The low er p ic tu r e i s th a t o f a c o n tro l tum or, r a te d 3+ h i s t o l o g i c a l l y . (M a g n ific atio n 440X). « F ig u re 5. M icrophotograpli o f turrior No. 109 Hypopiiysectoiiiized. T his tumor had a dark appearance of th e t o t a l mass and con ta in e d a sm all amount o f glycogen, (M a g n ific a tio n 440X). CHAPTER VI _ SUM M APy 1 . L i t t l e glycogen i s found i n maimnaiy-type carcinom a. 2. Evidence te n d s to show th a t th e glycogen i s more prom inent i n th e slow growing tum ors, 3. Glycogen te n d s to be c o n c e n tra te d i n th e rin d o r growing a re a o f th e tum or. 4 . E vidence d id not I n d ic a te t h a t th e heavy co n n ectiv e t i s s u e strom a o f t h i s tum or bore a r e l a t i o n s h i p to th e s c a r c i t y o f glycogen found. 5. The g ra n u le s o f glycogen vary i n s i z e from sm all s in g le g r a in s t o la r g e heavy d r o p l e t - l i k e g ra n u le s and a re u s u a lly lo c a te d c lo se to th e n u c le i o f th e la r g e e p i t h e l i a l c e l l s ; o c c a s io n a lly i n n e c r o tic a r e a s e s p e c ia lly th o s e p a tc h e s b eg in n in g i n th e r in d a re a and r a r e l y i n th e c o n n ec tiv e t i s s u e strom a. 6. Many tum ors c o n tain ed a l i q u i d c e n te r due to n e c r o s is , which i s c o n tra ry to th e f in d in g s o f E a rle (1935) who claim ed t h a t a lth o u g h th e c e n te r of th e tumor was n e c r o tic , i t was firm . T h is le a d s to th e h y p o th e sis t h a t th e v a r i a t i o n i n glycogen fin d in g s between th e chem ical and h i s t o l o g i c a l m ight be due to t h i s l i q u e f a c t i o n . 7. Chemical e s tim a tio n , w h ile showing a s l i g h t in c re a s e i n g ly cogen f o r th e hypophysectom ized, does not show t h a t hypophysectomy allo w s f o r a g r e a te r c o n c e n tra tio n o f glycogen i n th e o p e ra ted as a g a in s t th e in - 39 t a c t a n im a ls . T h is i s c o n tra ry t o th e f in d in g s o f S c h o tt, Sam uels and B a ll (1 9 3 7 ). 8 . C e r ta in o f th e tu m o rs w hich showed a d a rk a p p e a ra n c e th ro u g h o u t th e m ass seemed t o he lo w e r i n g ly c o g e n c o n te n t th a n th e u s u a l w h ite , tr a n s l u c e n t tu m o rs o f a p p ro a c h in g s i z e and w e ig h t. 9 . The s m a lle r tu m o rs, a s a g ro u p , seemed t o c o n ta in more g ly co g en th a n th e l a r g e r o n e s . BIBLIOGRAPHT Books B a rk e r, E l l i s J . , C a n c e r. The S u rg eo n and th e R e s e a r c h e r. London: Jo h n M urray, 1928. Ew ing, J a m e s ., C a u s a tio n . D ia g n o sis and T rea tm e n t o f C an c e r. . B a ltim o re : W illia m s and W ilk in s Co. 1931. H a d fie ld , G e o f f r e y ., and Law rence P . G e r r o d ., R ecen t A dvances i n P a th o lo jô r. P h il a d e lp h ia : P . B la k is to n ’ s Son and Co. I n c . 1952. M acCallum , W. G ., A T extbook o f P a th o lo g y . P h ila d e lp h ia and London: W. B. S a u n d e rs Co. 1937. Maximow, A le x an d e r A ., and W illia m B loom ., A T extbook o f H is to lo g y . P h ila d e lp h ia and London: W. B. S au n d ers Co. 1937. W illia m s , W. R o g e r ., The N a tu ra l H is to ry o f C a n c e r. W illia m Wood and Co. 1908. A r t i c l e s B abes, A ., and L a z a r e s c u - P a n tz u ., ’’C ancer c e l l s r i c h i n g ly c o g e n , i n c e r v i x - u t e r i . ” V irchow s A rc h . F . p a th . O n s t ., 2 7 5 :7 0 5 -8 4 , 1930. B a l l , H. A ., L . T . Sam uels and W. Sim pson, " E f f e c t o f hypophysectom y on t r a n s p l a n t a b l e mammary carcin o m a i n th e w h ite r a t . " Am. 2 . C an c e r. 16: 3 5 1 -9 , 1932. ■ and L . T . S am u els, " E f f e c t o f i^pophysectom y on au to g en o u s tu m o rs ." Am. %. C a n c e r. 26: 5 4 7 -5 1 , 1936. B ern h ard , F . , " D e te rm in a tio n o f g ly c o g e n i n b e n ig n and m a lig n a n t tum or c e l l s . " K lin M nhnschr, I : 1 1 8 4 -8 5 , 1928. B o rg h i, B ., "Amount o f g ly c o g en found i n aden o carcin o m a i n r a t . " B o ll. 2* S o c. i t a l d i b l o l . s p a r . -3: 5 7 8 -8 2 , 1928. E a r le , W. R . , "S tu d y o f W alker R at Mammary C a n c e r." Am. 2* C an cer, 2 4 : 256, 1935. Emge, L . A ., and K . M. M urphy, " E f f e c t o f I^pophysectom y and p i t u i t a r y grow th hormone on tr a n s p la n t e d r a t sarco m a ." Am. 2» O b s te t. and G ynec; . 32: 5 9 3 -6 0 9 , 1936. F a ro y , G ., "G lycogen in developm ent o f tu m o rs, norm al t i s s u e and v a r io u s ty p e s o f o rg a n is m s ." R ev, c r l t . de p a th , e t de th e r a p . . 2 / 9 4 3 -4 7 , 1931. G o ld fe d o r, A ., "G lycogen i n tum or t i s s u e . " Am. J . C a n c e r., 18: 6 4 4 , 1 9 3 3 . Lund, H ., " E x is te n c e o f s p e c i f i c g ly c o g e n p e c u l i a r t o tum or and em bryonic t i s s u e . " Compte, re n d u , s e e , de b l o l . . 110, 1 1 2 1 -2 4 , 1932. McEuen, C. S . , and D. L . Thompson, " E f f e c t o f hypophysectom y on groivth o f W alker r a t tu m o r." B r i t . 2* E x p e r. P a th . . 14: 3 8 4 -9 1 , 1955. P r o c . s e e . E x p er. B io l, and Med. . 30, 9 2 8 -2 9 , 1933. McEuen, C. S . , and D. L . Thompson, "G row th o f W alker r a t tu m o r i n young and o ld r a t s . " B r i t . 2* ]Exper. P a th . . 15, 1933. P a s t e l s , S . , and G. L eo n ard , "T ech n ic o f p re p a rin g h i s t o l o g i c s e c tio n s ^ t o d e t e c t g ly co g en i n t i s s u e s . " B u l l , d ’ h i s t o l . a p p l i g . ^ l a p h y s io l. , 1 2 :2 9 3 -9 9 , 1935. P e tro w a , E . , "G lycogen c o n te n t o f myomatous tum ors o f u t e r u s . " Z e n t r a l b l . 2* Gypok. , 56: 1 0 4 5 -4 8 , 193 2 . R ap p in , G ., "R o le o f g ly c o g e n s is i n o r i g i n end e v o lu tio n o f c a n c e r ." B u ll. A sso c , f r a n c . 1 ’ e tu d e du c a n c e r. .2 1 : 4 8 9 -9 1 , 1932 Ryoj u i n , K in o s ita and D aikoku, K o s iro , "A new m ethod f o r h i s t o l o g i c a l d e te r m in a tio n o f g ly c o g e n ." T ra n . S o c. P a th . J a p . . 26: 1 2 7 -3 0 , 1936.

Linked assets

University of Southern California Dissertations and Theses

Conceptually similar

PDF

Some histological considerations of the aortic paraganglionic bodies (of Zuckerkandl) of the guinea pig

PDF

A comparative histological study of the kidneys of the lower vertebrates

PDF

A comparative study of the histological structure of oral glands on certain mammals

PDF

Histological variations in the walls of the digestive tract of salamanders

PDF

Classification and observation of Formicidae in the Los Angeles vicinity

PDF

The microscopic anatomy of seven genera of Ascaroidea and one genus of Spiruroidea

PDF

Embryologic and histologic studies of the human thymus

PDF

Comparative study: Swedish neutrality and Egyptian non-alignment

Cytological and genetical study of certain regions of the chromosomes in <italic>Drosophila pseudoobscura</italic>

PDF

Cytological and genetical study of certain regions of the chromosomes in Drosophila pseudoobscura

PDF

Histological changes induced in the harderian gland of the mouse by transplantation in the anterior chamber of the guinea pig eye

A seasonal study of the interstitial cells of the testes of the Brewer Blackbird (<italic>Euphagus cyanocephalus</italic>)

PDF

A seasonal study of the interstitial cells of the testes of the Brewer Blackbird (Euphagus cyanocephalus)

PDF

A study of the Massasauga as found in Trumbull County, Ohio

PDF

A neurochemical and behavioral analysis of the basal ganglia

A study of metabolic exchanges in <italic>Citellus tridecemlineatus</italic> (Mitchell) at different temperature

PDF

A study of metabolic exchanges in Citellus tridecemlineatus (Mitchell) at different temperature

PDF

Influence of a liquid diet on water and electrolyte equilibrium and nitrogen excretion in the albino rat

PDF

A study of the fish of Anaheim Slough with reference to classification and nature of their food

PDF

Studies in metabolism: Oxygen consumption in certain lizards

A microscopical study of the reproductive organs of <italic>Moniezia expansa</italic>, a cestode

PDF

A microscopical study of the reproductive organs of Moniezia expansa, a cestode

PDF

Mites parasitic in the lungs and air passages of seals, with special emphasis on their occurence in the California sea lion

PDF

A quantitative study of marine phytoplankton from the Hancock Pacific expeditions of 1933 and 1937

Asset Metadata

Creator Hall, W. L. (author)

Core Title A study of the histological distribution of glycogen in tumors of hypo-physectomized and non-hypophysectomized rats

Contributor Digitized by ProQuest (provenance)

Degree Master of Arts

Publisher University of Southern California (original), University of Southern California. Libraries (digital)

Tag Biological Sciences,OAI-PMH Harvest

Format application/pdf (imt)

Language English

Permanent Link (DOI) https://doi.org/10.25549/usctheses-c39-256310

Unique identifier UC11315893

Identifier EP67126.pdf (filename),usctheses-c39-256310 (legacy record id)

Legacy Identifier EP67126.pdf

Dmrecord 256310

Document Type Thesis

Format application/pdf (imt)

Rights Hall, W. L.

Type texts

Source University of Southern California (contributing entity), University of Southern California Dissertations and Theses (collection)

Access Conditions The author retains rights to his/her dissertation, thesis or other graduate work according to U.S. copyright law. Electronic access is being provided by the USC Libraries in agreement with the au...

Repository Name University of Southern California Digital Library

Repository Location USC Digital Library, University of Southern California, University Park Campus, Los Angeles, California 90089, USA