ENVIRONMENTAL
MUTAGENESIS
Off icial Journal of the
Environmental
Mutagen
Society
Volume 2,1980
Seymour Abrahamson
EDITOR-IN-CHIEF
Alan R. Liss, Inc., New York
© 1 9 8 0 A l a n R . Liss, Inc.
150 F i f t h A v e n u e
New York, N Y
10011
ENVIRONMENTAL
M UTAGENESIS
Official Journal of the Environmental Mutagen Society
PRESENT OFFICERS, ENVIRONMENTAL
President
M U T A G E N SOCIETY, 1979-1980
President-El ect
Secretary
Treasurer
S. Wolff
S. Green
A . D . Mitchell
M . L . Mendelsohn
Councillors
R. Albertini
R . H . Haynes
M . J . Prival
M. Shelby
A . D . Bloom
M. Hite
V . A . Ray
V . Simmon
D J . Brusick
R. Kimball
H.S. Rosenkranz
D. Stoltz
V . C . Dunkel
J . McCann
L.B. Russell
R. Valencia
W.G. Flamm
D. Matheson
G . Sega
EDITOR-IN-CHIEF
Seymour Abrahamson, University of Wisconsin, Madison, Wisconsin
ASSOCIATE EDITOR
Carter Denniston, University of Wisconsin, Madison, Wisconsin
BOOK REVIEW EDITOR
Benjamin P. Sonnenblick, Rutgers University, Newark, New Jersey
R E F E R E N C E EDITORS
Elizabeth S. Von Halle, Oak Ridge National Laboratory, Oak Ridge, Tennessee
John S. Wassom, Oak Ridge National Laboratory, Oak Ridge, Tennessee
Bradford L. Whitfield, Oak Ridge National Laboratory, Oak Ridge, Tennessee
CONTRIBUTING EDITORS
Bruce N. Arnes, University of California, Berkeley, California
Verne A. Ray, Pfizer Pharmaceuticals, G r o t o n , Connecticut
Frederick J . de Serres, National Institute of Environmental Health Sciences, Research Triangle
Park, North Carolina
EDITORIAL B O A R D
J. Grant Brewen, A l l i e d Chemical C o r p o r a t i o n , M o r r i s t o w n , New Jersey
Herman E. Brockman, Illinois State University, N o r m a l , Illinois
David Brusick, Litton Bionetics, Inc., Kensington, Maryland
Anthony V . Carrano, Lawrence Livermore Laboratory, Livermore, California
Donald Clive, Burroughs Wellcome C o . , Research Triangle Park, North Carolina
John W. Drake, National Institute of Environmental Health Sciences, Research Triangle Park,
North Carolina
Philip E. Hartman, Johns Hopkins University, Baltimore, Maryland
Robert H. Haynes, Y o r k University, T o r o n t o , Ontarlo
William R. Lee, Louisiana State University, Baton Rouge, Louisiana
Marvin S. Legator, University of Texas MedicahBranch, Galveston, Texas
Michael W. Lieberman, Washington University School of Medicine, St. Louis, Missouri
Joyce McCann, University of California, Berkeley, California
Michael Plewa, University of Illinois, Urbana, Illinois
Louise Prakash, University of Rochester School of Medicine, Rochester, New Y o r k
R. Julian Preston, Oak Ridge National Laboratory, Oak Ridge, Tennessee
Liane B. Russell, Oak Ridge National Laboratory, Oak Ridge, Tennessee
Robert Shapiro, New Y o r k University, New Y o r k , New Y o r k
Bernard Strauss, University of Chicago, Chicago, Illinois
Sheldon Wolff, University of California, San Francisco, California
Stanley Zimmering, B r o w n University, Providence, Rhode Island
T h e code at t h e b o t t o m of t h e first page of each article in this Journal indicates t h e C o p y r i g h t o w n e r ' s c o n s e n t t h a t
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Contents
Volume 2, Number 1
1980
EDITORI A L
Editor's Prologue
Seymour Abrahamson
INVITED
1
REVIEW
B a c t e r i a l M u t a g e n e s i s : R e v i e w o f N e w Insights
Philip E . Hartman
RESEARCH
3
PAPERS
Tissue-Specific I n d u c t i o n o f Sister C h r o m a t i d Exchanges by E t h y l
Carbamate in Mice
G l a d w i n T. R o b e r t s and J a m e s W . A l l e n
17
Effects of E p o x i d e Hydratase Inhibitors in F o r w a r d and Reversion
Bacterial Mutagenesis A s s a y S y s t e m s
D e r e k G u e s t and J o h n G . D e n t
27
S i s t e r C h r o m a t i d E x c h a n g e I n V i v o i n M i c e : I. T h e I n f l u e n c e o f I n c r e a s i n g
Doses o f B r o m o d e o x y u r i d i n e
J a m e s L . W i l m e r a n d E . R . S o a r e s . . .• . . . *.\ . . . ^ .
.,
35
S e x - R e l a t e d D i f f e r e n c e s i n C y t o g e n e t i c E f f e c t s o f B e n z e n e i n the B o n e
M a r r o w of Swiss M i c e
Julianne Meyne and M . S . Legator
43
R e g u l a t i o n of D i m e t h y l n i t r o s a m i n e M e t a b o l i s m by A n d r o g e n i c
Hormones
K. Bilkshi, D . B r u s i c k , L . P . B u l l o c k , and C . W . B a r d i n
51
M u t a g e n i c ü y o f 2- a n d 3 - C a r b o n H a l o g e n a t e d C o m p o u n d s i n the
S a l m o n e l l a / M a m m a l i a n - M i c r o s o m e Test
S.J. Stolzenberg and C . H . H i n e
59
M i t o t i c Arrest by B e n z i m i d a z o l e A n a l o g s in H u m a n L y m p h o c y t e
Cultures
H e n r y E. Holden, Paula A . C r i d e r , and Margitta G , Wahrenburg
67
I. B a c t e r i a l M u t a g e n i c i t y o f P a r t i c u l a t e s F r o m H o u s t o n A i r
B a r b a r a Lee B o r n s P r e i d e c k e r
75
II. C o m p a r a t i v e E x t r a c t i o n o f H o u s t o n A i r P a r t i c u l a t e s W i t h C y c l o h e x a n e
or a M i x t u r e of B e n z e n e , M e t h a n o l , a n d
Dichloromethane
B a r b a r a Lee B o r n s P r e i d e c k e r
85
M u t a g e n i c Effects o f B l e o m y c i n i n D r o s o p h i l a m e l a n o g a s t e r
H. Traut
MEETING
89
REPORT
S e c o n d European W o r k s h o p o n Bacterial In V i t r o M u t a g e n i c i t y Test Systems
J . P . S e i l e r , I.E. M a t t e r n , M . H . L . G r e e n , a n d D . A n d e r s o n
97
Volume 2, Number 2
RESEARCH
1980
PAPERS
Mutagenicity of Effluents F r o m an E x p e r i m e n t a l F l u i d i z e d Bed Coal C o m b u s t o r
Charles R. Clark and Charles H . H o b b s
101
L a c k o f an I n d i c a t i o n o f M u t a g e n i c Effects o f D i n i t r o t o l u e n e s and
Diaminotoluenes in Mice
E . R . Soares a n d L . F . L o c k
111
I n c r e a s e s i n M o r p h o l o g i c a l l y A b n o r m a l S p e r m i n R a t s E x p o s e d to
Co
6 0
Irradiation
L . F . L o c k a n d E . R . Soares
125
The Mutagenic Effect of P l a t i n u m C o m p o u n d s in D r o s o p h i l a melanogaster
R . C . Woodruff, R. V a l e n c i a , R . F . L y m a n , B . A . Earle, and J.T. B o y c e
133
In V i t r o Induction o f Segregational Errors o f Chromosomes by Natural
Cannabinoids in Normal H u m a n L y m p h o c y t e s
Richard T. H e n r i c h , T a k a y u k i N o g a w a , and A k i r a M o r i s h i m a
139
X I r r a d i a t i o n a n d Sister C h r o m a t i d E x c h a n g e i n C u l t u r e d H u m a n L y m p h o c y t e s
W i l l i a m F . Morgan and Peter E . Crossen
149
Sister C h r o m a t i d E x c h a n g e S t u d i e s i n H u m a n F i b r o b l a s t - R a t
Hepatocyte
C o - C u l t u r e s : A N e w I n V i t r o S y s t e m to S t u d y S C E s
A n d r e w D . K l i g e r m a n , Stephen C . S t r o m , and George M i c h a l o p o u l o s
157
E n z y m e Mutants Induced by L o w - D o s e - R a t e 7-Irradiation in D r o s o p h i l a :
Frequency and Characterization
R o b e r t R. Racine, Charles H . L a n g l e y , and Robert A . V o e l k e r
167
Genetic Effects of Strong Magnetic Fields i n D r o s o p h i l a melanogaster:
II. L a c k o f I n t e r a c t i o n B e t w e e n H o m o g e n e o u s F i e l d s a n d F i s s i o n
Neutron-Plus-Gamma Radiation
P . G . Kaie and J.W. B a u m
179
a
E v i d e n c e T h a t the R e p a i r D e f i c i e n t mei-9
Female in D r o s o p h i l a melanogaster
Is a S t r o n g P o t e n t i a t o r o f C h r o m o s o m e L o s s I n d u c e d i n t h e P a t e r n a l
Genome by Dimethylnitrosamine
S. Z i m m e r i n g , A . W . H a r t m a n n , a n d S . F . C o o p e r
187
T h e A c t i o n o f T h r e e A n t i c l a s t o g e n s 011 the I n d u c t i o n o f S i s t e r C h r o m a t i d
Exchange by T r e n i m o n and 8 - H y d r o x y q u i n o l i n e Sulfate in H u m a n
L y m p h o c y t e Cultures
E. Gebhart and H . K a p p a u f
BOOK AND ARTICLE
REVIEWS
B.P. Sonnenblick
MEETING
191
201
REPORT
E l e v e n t h A n n u a l M e e t i n g o f the E n v i r o n m e n t a l Mutagen S o c i e t y
O f f i c e r s o f the S o c i e t y
203
Councilors
203
Program Committee
204
Registration Fees and H o u r s
204
General Information
204
Program
206
Abstracts
230
A u t h o r I n d e x to A b s t r a c t s
317
Announcement
321
1980
Volume 2, Number 3
EDITORIAL
L a b o r a t o r y Safety and H a n d l i n g Procedures for C h e m i c a l Mutagens
David J. Brusick
RESEARCH
323
PAPERS
V a r i a t i o n i n the B a s e l i n e Sister C h r o m a t i d E x c h a n g e
Frequency
in H u m a n L y m p h o c y t e s
A . V . C a r r a n o , J . L . M i n k l e r , D . G . S t e t k a , a n d D . H . M o o r e II
325
I n d u c t i o n of C h r o m o s o m e Shattering and M i c r o n u c l e i b y Ultraviolet L i g h t
a n d C a f f e i n e . I. T e m p o r a l R e l a t i o n s h i p a n d A n t a g o n i s t i c E f f e c t s o f t h e
Four
Deoxyribonucleosides
C. C r e m e r , T . C r e m e r , a n d M . S i m i c k o v a
339
T h e Anaerobe-Mediated Mutagenicity of 2-Nitrofluorene and 2-Aminofluorene
for Salmonella t y p h i m u r i u m
G e o r g e E . K a r p i n s k y a n d H e r b e r t S. R o s e n k r a n z
353
M u t a g e n i c i t y o f Pesticides Evaluated by Means o f G e n e - C o n v e r s i o n i n
S a c c h a r o m y c e s cerevisiae a n d i n A s p e r g i l l u s n i d u l a n s
M . de B e r t o l d i , M . G r i s e l l i , M . G i o v a n n e t t i , a n d R . B a r a l e
359
Absence of Arsenite Mutagenicity in E coli and Chinese Hamster Cells
T . G . Rossman, D . Stone, M . Molina, and W. T r o l l
371
U s e o f H y d r o x y u r e a i n the M e a s u r e m e n t o f D N A R e p a i r b y the
B N D Cellulose M e t h o d
J a m e s I r w i n and B e r n a r d Strauss
381
Sunlight-Induced Mutagenesis and T o x i c i t y in L 5 1 7 8 Y Mouse Cells:
Determination and Comparison With Other Light Sources
K e n n e t h K r e l l and E l i z a b e t h D . Jacobson
389
E f f e c t o f N - A l k y l C h a i n L e n g t h o n the M u t a g e n i c i t y o f N - N i t r o s a t e d
1-Naphthyl N-Alkylcarbamates
B r y a n K . E y a and R o n a l d E . Talcott
395
Mutagenicity Tests o f Fabric-Finishing Agents i n S a l m o n e l l a t y p h i m u r i u m :
Fiber-Reactive W o o l Dyes and C o t t o n F l a m e R e t a r d a n t s
James T. M a c G r e g o r , M a r t i n J . D i a m o n d , L a u r e n c e W. M a z z e n o , Jr.,
and Mendel F r i e d m a n
405
E s t i m a t i o n of the Weight-Dependent P r o b a b i l i t y o f D e t e c t i n g a M u t a g e n
W i t h the A r n e s A s s a y
James B. Johnston and Philip K . H o p k e
BRIEF
419
COMMUNICATION
Genes C o n t r o l l i n g Sensitivity to A l k y l a t i o n and X - R a y Damage o n
C h r o m o s o m e 3 o f D r o s o p h i l a melanogaster
A . K . Beck, R . R . Racine, and F . E . Würgler
MEETING
425
REPORT
M e e t i n g R e p o r t o n the S e c o n d E M S W o r k s h o p
David J. Brusick
Program Announcement
431
433
1980
Volume 2, Nuniber 4
RESEARCH
PAPERS
Differential I n d u c t i o n of Sister C h r o m a t i d E x c h a n g e s by I n d i r e c t - A c t i n g MutagenC a r c i n o g e n s at E a r l y a n d L a t e Stages o f E m b r y o n i c D e v e l o p m e n t
L o r i A . T o d d and Stephen E. B l o o m
How
435
M a n y L o c i o n the X - C h r o m o s o m e o f D r o s o p h i l a m e l a n o g a s t e r C a n M u t a t e
to R e c e s s i v e L e t h a i s ?
S. A b r a h a m s o n , F . E . W ü r g l e r , C . D e J o n g h , a n d H . U n g e r M e y e r
Short-Term Cytogenetic Assays of Nine Cancer Chemotherapeutic
447
Drugs With
Metabolie Activation
William W. A u , Dennis A . J o h n s t o n , C h e r y l C o l l i e - B r u y e r e , and T . C . Hsu
455
H y p e r t h e r m i a I n d u c e d D i s s o c i a t i o n o f the X - Y B i v a l e n t i n M i c e
M.L.
Garriott and C . L . Chris man
465
T h e R e l a t i v e C o n t r i b u t i o n s o f B a n d T L y m p h o c y t e s i n the H u m a n P e r i p h e r a l
B l o o d M u t a g e n T e s t S y s t e m as D e t e r m i n e d b y C e l l S u r v i v a l , M i t o g e n i c S t i m u l a t i o n ,
and Induction of Chromosome Aberrations by Radiation
Jeffrey L . S c h w a r t z and Mary Esther G a u l d e n
473
A C o m p a r i s o n o f the A b i l i t y o f F r o g a n d R a t S-9 t o A c t i v a t e P r o m u t a g e n s
in the A r n e s T e s t
Albert M . Cheh, Alan B. Hooper, Jill Skochdopole, Craig A . Henke,
and Robert G . M c K i n n e l l
487
Clastogen-Induced Micronuclei in Peripheral B l o o d Erythrocytes:
T h e Basis o f
an I m p r o v e d M i c r o n u c l e u s T e s t
James T . M a c G r e g o r , Carol M . Wehr, and Daniel H . G o u l d
509
P o t e n t i a t i o n o f C h r o m o s o m e L o s s I n d u c e d i n the P a t e r n a l G e n o m e b y M e t h y l
Methanesulfonate and Procarbazine F o l l o w i n g Matings With Repair
Deficient me/-9
a
Females of Drosophila
S. Z i m m e r i n g a n d K . L .
Kammermeyer
515
T h e E v a l u a t i o n o f the E p o x i d e D i l u e n t , n - B u t y l g l y c i d y l E t h e r , i n a Series o f
Mutagenicity Assays
T.H.
C o n n o r , J . B . W a r d , Jr., J . M e y n e , T . G . P u l l i n , a n d M . S . L e g a t o r
G e n o t o x i c A c t i v i t y in Microorganisms of T e t r y l , 1,3-Dinitrobenzene
521
and
1,3,5-Trinitrobenzene
D o u g l a s B . M c G r e g o r , C o l i n G . R i a c h , R o w a n M . Hast w e l l , a n d J a c k C . D a c r e
BRIEF
531
COMMUNICATION
d
A M a t e r n a l E f f e c t i n H o m o z y g o u s mei'9'
mei-4p^
Repair Deficient Drosophila
m e l a n o g a s t e r F e m a l e s I n f l u e n c i n g the R e c o v e r y R a t e o f P r o g e n y B e a r i n g
a Y Chromosome
S. Z i m m e r i n g a n d S . F . C o o p e r
BOOK A N D ARTICLE
B.P.
Sonnenblick
543
REVIEWS
547
A u t h o r Index
549
Subject Index
551
E n v i r o n m e n t a l Mutagenesis 2 : 3 3 9 - 3 5 1
(1980)
Induction of Chromosome Shattering and
Micronuclei by Ultraviolet Light and
Caffeine. I. Temporal Relationship and
Antagonistic Effects of the Four
Deoxyribonucleosides
C. Cremer, T. Cremer, and M. Simickova
I n s t i t u t e o f Human Genetics,
University
o f Freiburg,
A i b e r t s t r . 11, D - 7 8 0 0 Freiburg
i.
B r . (C.C., M . S . ) , and I n s t i t u t e o f A n t h r o p o l o g y a n d Human Genetics,
University
of
Heidelberg,
I m Neuenheimer
F e l d 3 2 8 , D - 6 9 0 0 Heidelberg
( T . C J , Federal R e p u b l i c o f
German y
It is k n o w n t h a t n u c l e o s i d e s m a y have a n t i m u t a g e n i c a n d a n t i c l a s t o g e n i c effects.
H e r e , w e have i n v e s t i g a t e d t h e i n f l u e n c e o f n u c l e o s i d e s o n t h e i n d u c t i o n o f
shattered c h r o m o s o m e s (fragmentation a n d / o r pulverization of c h r ö m o s o m e s of
a m i t o t i c cell) a n d o f m i c r o n u c l e i b y u l t r a v i o l e t ( U V ) l i g h t a n d c a f f e i n e . A s y n chronous cell cultures o f a V 7 9 subline o f the Chinese hamster were irradiated
at w a v e l e n g t h 2 5 4 n m u s i n g f l u e n c e s u p t o 5.2 j o u l e s / n r . F o l i o w i n g Irradiat i o n , t h e cells w e r e p o s t i n c u b a t e d e i t h e r w i t h 1.0 m M o r 2 . 0 m M c a f f e i n e a l o n e
or w i t h c a f f e i n e p l u s t h e f o u r d e o x y r i b o n u c l e o s i d e s ( d X s ) ( c o n c e n t r a t i o n 0.1
m M each). A f t e r different i n c u b a t i o n times (three to 2 4 hours), c h r o m o s o m e
p r e p a r a t i o n s w e r e p e r f o r m e d . I n o t h e r e x p e r i m e n t s , s y n c h r o n i z e d cells w e r e
used. T h e percentage o f metaphase spreads w i t h shattered c h r o m o s o m e s and the
p e r c e n t a g e o f cells w i t h m i c r o n u c l e i w e r e d e t e r m i n e d . P o s t - t r e a t m e n t w i t h
caffeine a l o n e r e s u l t e d i n s h a t t e r e d c h r o m o s o m e s i n a h i g h p e r c e n t a g e o f cells
at t h e first p o s t - i r r a d i a t i o n m i t o s i s as d e s c r i b e d p r e v i o u s l y . F o r m a t i o n o f cells
w i t h m i c r o n u c l e i was o b s e r v e d o n l y after t h e a p p e a r a n c e o f m i t o t i c cells w i t h
s h a t t e r e d c h r o m o s o m e s , t h e m a x i m u m p e r c e n t a g e o f cells w i t h m i c r o n u c l e i
being smaller t h a n the m a x i m u m percentage o f cells w i t h shattered c h r o m o s o m e s . T h e s t r o n g p o t e n t i a t i n g effect o f U V - l i g h t p l u s c a f f e i n e was s i g n i f i cantly r e d u c e d , h o w e v e r , if the post-treatment was p e r f o r m e d w i t h caffeine plus
n u c l e o s i d e s . A s i g n i f i c a n t r e d u c t i o n was a l s o o b s e r v e d i n t h e p e r c e n t a g e o f
m i c r o n u c l e i . A n evaluation o f the m i t o t i c indices and o f cell-cycle Parameters
i n d i c a t e s t h a t t h e effect o f n u c l e o s i d e s w a s n o t d u e t o e n h a n c e d i n t e r p h a s e
death.
2
Key
words: chromosome shattering, ultraviolet (UV)
micronuclei
light, caffeine, nucleosides, antimutagens,
INTRODUCTION
In a n u m b e r o f c e l l s t r a i n s , e s p e c i a l l y i n r o d e n t c e l l s , c a f f e i n e is k n o w n t o p o t e n t i ate the c h r o m o s o m e d a m a g i n g effects o f u l t r a v i o l e t ( U V ) l i g h t a n d a n u m b e r o f c h e m i c a l
mutagens [ K i h l m a n , 1 9 7 4 ; N i l s s o n a n d L e h m a n n , 1 9 7 5 ; H a r t l e y - A s p , 1 9 7 6 ; K i h l m a n , 1 9 7 7 ;
Received January 2, 1980; accepted M a y 8 , 1 9 8 0 .
Paits o f this investigation w i l l be presented i n the doctoral dissertation of M . Simickova to be submitted
to the F a c u l t y of Medicine, University o f Freiburg i . B r .
0 1 9 2 - 2 5 2 1 / 8 0 / 0 2 0 3 - 0 3 3 9 $ 0 2 . 6 0 © 1980 A l a n R . Liss, Inc.
340
Cremer, Cremer, and Simickova
Roberts, 1978]. A striking phenomenon observed after UV irradiation (X = 254 nm) and
caffeine post-treatment is the frequent occurrence of cells with generalized chromosome
shattering (GCS) (fragmentation and/or pulverization of all chromosomes of a mitotic cell)
[Nilsson and Lehmann, 1975; T. Cremer et al, 1979].
In the present investigation, we have studied the influence of nucleosides on this
phenomenon. These substances are known to have antimutagenic and anticlastogenic effects
[Novick and Szilard, 1952; Kihlman, 1977; Gebhart, 1977]. Here, it is shown that the addition of deoxyribonucleosides to the postirradiation medium exerts a strong antagonistic
effect on the induction of GCS. In addition, we examined the influence of nucleosides on
the production of cells with micronuclei [Boller and Schmid, 1970; Countryman and Heddle,
1976]. The presence of nucleosides also reduced the percentage of micronuclei produced by
UV light and caffeine.
MATERIAL AND METHODS
Cell Material and Culture Conditions
Cells of a subline of V79 Chinese hamster cells [Cremer et al, 1976] were used. This
cell line has a modal chromosome number of 21 and a mean generation time of 13—14
hours. The cells were grown in Eagle's minimum essential medium (MEM) supplemented
with 10% fetal calf serum (FCS), nonessential amino acids, and antibiotics (100 units/ml
penicillin and 100 Mg/ml streptomycin) in a humidified atmosphere with 5% C 0 . MEM
(Flow Laboratories, Germany) was free from deoxyribonucleosides.
For experiments, cells were grown in 6-cm plastic petri dishes (Nunc/Denmark).
Asynchronous cultures were inoculated after trypsination and grown two days before use.
Synchronous populations were obtained by the mitotic shake-off procedure which resulted in at least 90% mitotic cells as revealed by direct chromosome preparations. The
percentage of S-phase cells in these populations was small (< 2%) as shown by pulse labeling with H-thymidine prior to detachment.
2
3
U V Irradiation and Post-treatment
3
Asynchronous cells grown in petri dishes were labeled with H-thymidine (0.1
/uCi/ml, 5 juCi/mmol; Amersham Buchler) for 30 minutes prior to irradiation. Synchronized cells were used without prelabeling. Medium was removed before irradiation and
the cells were washed twice with Hanks' Solution (without phenol red). Thereafter, cells
were irradiated from above, while covered with a 1 mm layer of Hanks' Solution. A
germicidal lamp (Sterisol 5143, Original Hanau) was used emitting predominantly at 254
nm wavelength. The fluence rate was determined to be 3.5 W/m by means of a calibrated
photodiode (United Detector Technology, Santa Monica, California). The duration of irradiation (0.75 seconds, 1.5 seconds) was controlled by openings in rotating masks above
the cells which were moved by the wheel of a record player. This procedure allowed a precise adjustment of the irradiation time. Immediately after the irradiation, the cells were
postincubated at 37°C in MEM with 10% FCS, either with caffeine (1-2 mM) alone or with
caffeine plus the four deoxyribonucleosides (dXs) (deoxyadenosine, deoxycytidine, deooxyguanosine, and thymidine; concentration 0.1 mM each). Except for the addition of dXs,
all treatments were identical and made in duplicate. After different incubation times (3—
27 hours), in situ chromosome preparation was performed [Zorn et al, 1976]. Colchicine
2
Anticlastogenic Effects o f Nucleosides
341
(2 Mg/ml) was added 3 hours before preparation. The dishes were air-dried and stained with
aceto-orcein. The scoring of dishes was performed "blind," ie, in the absence of Information
about the treatment regimen. To examine chromosome damage and micronucleus formation , at least 50 metaphases and 500 interphase cells, respectively, were analyzed per dish;
ie, at least 100 metaphasefiguresand 1,000 interphase cells were scored for each treatment
and fixation time.
For Statistical evaluation the binomial assumption was made, and the confidence limits
Pi ^ p ^ P2 for proportions were determined [Beyer, 1974]. The observed frequency is p
and Pi and p are the lower and upper limits of the 95% confidence interval, respectively.
Throughout the text, the 95% confidence intervals are given as decimals. The actual data
are expressed as a percent. The term "nonsignificant difference" between two observed frequencies pj, pjj means that the 95% confidence intervals of pj and p overlapped; if the
term "signficant difference" is used, the 95% confidence intervals of pj and pjj were clearly
separated from each other.
2
T1
Autoradiography
After microscopic evaluation as described above, pulse-labeled cells were covered with
Ilford nuclear emulsion K2 and processed following Standard procedures [Zorn, 1978]. Exposure time at 4°C was two weeks. In autoradiographs, the percentage of labeled metaphases
was determined.
RESULTS
1. Classification of Metaphase Figures
The metaphase figures obtained following whole-cell irradiation (X = 254 nm) of V79
cells and post-treatment with caffeine in the presence or absence of deoxyribonucleosides
were classified in the following way [Zorn et al, 1977; Zorn, 1978; T. Cremer et al, 1980a]:
class A: No recognizable alterations of chromosome morphology; class B: Metaphase plates
with one, occasionally two alterations. In most cases, these alterations were achromatic
lesions ("gaps") or chromatid breaks. Occasionally, chromatid exchange figures were found.
Deviations from the modal chromosome number were not classified as aberrations; class C:
All metaphase spreads containing more than two aberrations with the majority of chromosomes (11 and more) remaining intact, were classified as class C. In the whole-cell irradiation
experiments presented here, class C figures were a very rare event (< 1%), in contrast to experiments in which only a small part of the cell nucleus was UV- microirradiated [Zorn et al,
1977; Zorn, 1978]; class D: This class contains metaphase spreads in which the majority of
chromsomes (11 and more) showed an aberrant morphology, only one or several chromosomes remaining intact (Fig. la); class E: In metaphase spreads of class E, all chromosomes
were affected (GCS), appearing fragmented and/or pulverized (Fig. lb, c). In the majority,
figures as shown in Figure lc were observed. The term GCS is introduced as a descriptive
one to avoid any prejudice concerning the continuity or discontinuity of the DNA Strand.
Following UV- irradiation alone (up to 7 joules/m ) or caffeine post-treatment alone (1—2
mM), metaphase figures of classes C-E were rare events (< 1% each) [Zorn, 1978].
UV-irradiation plus caffeine post-treatment, however, induced GCS in a synergistic
(potentiating) way up to almost 100% of all metaphase figures obtained.
2
342
Cremer, Cremer, and Simickova
a
b
c
F i g . 1. Chromosome shattering following U V irradiation ( \ = 254 nm) o f V 7 9 cells and postincubation
with caffeine. a) Aberrant morphology i n the majority o f chromosomes, only one or several chromosomes
remaining intact: Class D ; b, c) A l l chromosomes affected (generalized chromosome shattering, G C S ) :
Class E ; fragmentation o f all chromosomes (b); pulverization o f all chromosomes (c).
2. Percentage of Cells with G C S
To investigate the influence of the addition of the dXs on the amount of GCS, cells
were treated under identical conditions except addition of dXs.
Figure 2 shows the result for asynchronous cells irradiated with 2.6 joules/m or
5.2 joules/m and post-treated with 1 mM caffeine with or without dXs. At 2.6 joules/m ,
the percentage of cells with GCS was low (<3%) in both cases (Fig. 2a, b), and no significant
difference was observed.
Irradiation with 5.2 joules/m and postincubation with 1 mM caffeine alone induced
a maximum of 33% (0.23 < p < 0.42) of cells with GCS (Fig. 2c). This percentage was
signficantly reduced, however, if dXs were added. In this case, a maximum of 12% (0.06
< p < 0.20) cells with GCS was obtained (Fig. 2d).
In Figure 3, the frequencies of metaphase figures classes A through E are shown for
cells post-treated with 2 mM caffeine. At this concentration, irradiation with 2.6 joules/m
and post-treatment with caffeine alone had a considerable effect: Up to 40% (0.30 < p
< 0.50) of cells with GCS were found (Fig. 3a). Again, the percentage of cells with GCS
was significantly reduced if dXs were added (Fig. 3b), the maximum being 12% (0.06 <
p < 0.20). The antagonistic effect of the addition of dXs was also observed following
irradiation with 5.2 joules / m . While a maximum of 97% (0.92 < p < 0.99) of cells with
GCS was observed without addition of dXs (Fig. 3c), the maximum was 78% (0.69 <
p < 0.86) in the presence of dXs.
In Figure 4, the results of experiments with synchronized cells are shown. These
cells were irradiated 3—5 hours after mitotic detachment and post-treated with 1 mM
caffeine in the presence or absence of dXs. Again, a significant reduction of cells with GCS
was obtained if nucleosides were added (Fig. 4a, b). After 15 hours incubation with 1 mM
caffeine in the absence of nucleosides, the amount of cells with GCS was 15% (0.08 <
p < 0.24) after irradiation with 2.6 joules/m and 50% (0.4 < p < 0.8) after 5.2 joules/m .
In the presence of dXs, the maximum percentages were 1.5% (0 < p < 0.07) and 3% (0
< p < 0.08) for the two doses, respectively. It is interesting to note that, in all cases with
a significant reduction of the percentage of cells with GCS, the presence of nucleosides also
significantly reduced the total aberration frequency (sum of percentages of cells with aberrations classes B through E, achromatic lesions excluded).
2
2
2
2
2
2
2
2
(%)
(%)
100
100
uv : 2.6 J/m2
uv
2.6 J / m
2
caffeine : 1 mM
caffeine : 1 mM
no
plus
dX.
50
dX
s
50
b)
Ja.
JBBL
18
2L
(hr)
(%)
o'
uv
100
V>
5.2 J / m *
O
Coffein* : 1 n
no
0Q
dX,
s.
o
50
2
c
c)
Fig. 2. U V irradiation (X = 254 nm) o f asynchronous V 7 9 cells and caffeine post-treatment (1 m M ) with
or without the four deoxyribonucleosides ( d X s ; concentration 0.1 m M each); Induction o f chromosome
alterations. Abscissa: Incubation time following irradiation (hr); ordinate: Percentage o f metaphase figures class A - E - • , no chromosomal alterations: Class A ; 3 , single defects (one, occasionally two aberrations) : Class B ; Q , aberrant morphology i n the majority o f chromosomes, only one or several chromosomes remaining intact: Class D ; B , all chromosomes affected (fragmentation and/or pulverization): generalized chromosome shattering ( G C S ) : Class E . (a) 2.6 j o u l e s / m , no d X s ; (b) 2.6 j o u l e s / m , plus
dXs. (c) 5.2 j o u l e s / m , no d X s , (d) 5.2 j o u l e s / m , plus d X s , F o r each value, at least 100 mitotic cells
were scored.
2
2
2
2
o
CO
Fig. 3. UV-irradiation (X = 254 nm) of asynchronous V 7 9 cells and caffeine post-treatment (2 m M )
with or without the four deoxyribonucleosides ( d X s ; concentration 0.1 m M each): Induction o f
chromosome alterations. Abscissa: Incubation time following irradiation (hr); Ordinate: Percentage
of metaphase figures classes A - E (see legend to F i g . 2) - • , class A ; 0 , class B ; 0 , class D ; B , class E .
(a) 2.6 j o u l e s / m , no d X s ; (b) 2.6 j o u l e s / m , plus d X s ; (c) 5.2 j o u l e s / m , no d X s ; (d) 5.2 j o u l e s / m ,
plus d X s . F o r each value, at least 100 mitotic cells were scored.
2
2
2
2
uv:
5.2 J /
(hr)
Fig. 4. UV-irradiation ( \ = 254 nm) o f synchronized V 7 9 cells ( G l and early S) and caffeine post-treatment (1 m M ) w i t h or
without the four deoxyribonucleosides (dXs; concentration 0.1 m M each): Induction o f generalized chromosome shattering
and formation o f micronucleated cells. a,b: Induction o f generalized chromosome shattering (GCS). Abscissa: Incubation time
following irradiation (hr); Ordinate: Percentage o f metaphase figures; classes A - E - o, no chromosomal alterations (class A ) ,
caffeine post-treatment without d X s ; • , no chromosomal alterations (class A ) , caffeine post-treatment in the presence o f d X s ;
• , all chromosomes affected (GCS), caffeine post-treatment without d X s ; • , all chromosomes affected (GCS), caffeine posttreatment in the presence of dXs. (a) 2.6 j o u l e s / m ; (b) 5.2 j o u l e s / m . F o r each value, at least 100 mitoses were scored. (c, d)
Induction o f cells with micronuclei: Abscissa: Incubation time following irradiation (hr); Ordinate: Percentage o f cells w i t h
micronuclei; o, no d X s ; • , plus d X s . (c) 2.6 j o u l e s / m ; (d) 5.2 j o u l e s / m . F o r each value, at least 1000 cells were scored.
2
2
2
2
(V.)
30
(%)
Coffeine
c ä f f t in«: 1mM
no
12
15
18
21
2k
dX
$
20
plus
12
(hr)
15
1«
21
24
dX
1 r
s
(hr)
3
o
7?
O
<
(%)
90
2mH
Coffeine :
20
plus
dX
(hr)
Fig. 5. UV-irradiation (X - 254 nm) of asynchronous V 7 9 cells and caffeine post-treatment with or
without the four deoxyribonucleosides (dXs; concentration 0.1 m M each): Induction of cells with
micronuclei. Abscissa: Incubation time following irradiation (hr); ordinale: Percentage o f cells with
m i c r o n u c l e i ; x , 0 j o u l e s / m ; 0 , 2.6 j o u l e s / m ;
5.2 j o u l e s / m . (a) 1 m M caffeine, no d X s ; (b) 1 m M
caffeine, plus d X s ; (c) 2 m M caffeine, no d X s ; (d) 2 m M caffeine, plus d X s . F o r each value, at least
1000 cells were scored. In the majority o f cases (70%), micronucleated cells had one to three micronuclei.
2
2
2
2 mM
s
Anticlastogenic Effects of Nucleosides
347
3. Formation of Micronuclei
The percentage of cells with micronuclei obtained after UV-irradiation (2.6 joules/
m and 5.2 joules/m ) without caffeine post-treatment or after caffeine treatment of unirradiated cultures was low (< 2%, 0.01 < p < 0.03), the differences being not significant.
A significant increase (up to 14%, 0.11 ^ p ^ 0.16), however, was observed following the
combined treatment with 5.2 joules/m and 1 mM caffeine (Fig. 5a). While 2.6 joules/m
and 1 mM caffeine post-treatment did not result in a significant increase of cells with
micronuclei (Fig. 5a), 2 mM caffeine post-treatment produced a considerable effect (maximum 16%, 0.14 < p < 0.18) even at this lower UV-dose (Fig. 5c). Addition of dXs significantly reduced the percentage of cells with micronuclei (Fig. 5b, d). The maxima of the
percentage of micronucleated cells obtained in the presence of dXs were 8% (0.06 < p <
0.10) for 5.2 joules/m plus 1 mM caffeine; 3.5% (0.02 < p < 0.045) for 2.6 joules/m
plus 2 mM caffeine; 13% (0.10 < p < 0.15) for 5.2 joules/m plus 2 mM caffeine (without dXs - 24% (0.21 < p < 0.26)).
A comparison of the data presented for the induction of GCS (Figs. 2 and 3) and
the results obtained for the production of micronuclei (Fig. 5) clearly shows that cells
with micronuclei appeared only after a significant increase of cells with GCS was observed.
For example, irradiation with 5.2 joules/m and 2 mM caffeine post-treatment resulted
in 60% mitotic cells with GCS after 9 hours incubation (Fig. 3c). At this time, the percentage of cells with micronuclei did not exceed the level of unirradiated controls.
A significant reduction of the percentage of cells with micronuclei by the addition
of dXs was observed also in case of synchronized cultures (Fig. 4c, d). A comparison
with the data presented in Fig. 4a, b shows that the formation of cells with GCS again
preceded the appearance of cells with micronuclei.
2
2
2
2
2
2
2
2
4. M i t o t i c Index and Cell-Cycle Parameters
For all incubation times, the mitotic indices were determined. It was found that the
addition of nucleosides did not significantly reduce the mitotic indices if compared with
cells treated in the absence of dXs. In some cases, the mitotic index was even slightly higher
when nucleosides were added. Furthermore, at 5.2 joules/m plus caffeine post-treatment
a reduction of the mitotic delay of approximately 3 hours was observed in the presence of
dXs.
The duration of S-phase and of G + prophase was estimated from the metaphase labeling index (MLI) curves according to the method of Evans and Scott [1964]. The results of
these estimates are presented in Table I. Up to UV fluences of 2.6 joules/m , the differences
in the duration of S-phase and G + prophase were small to nonexistant whether nucleosides were added or not. At 5.2 joules/m and postincubation with 1 mM and 2 mM caffeine,
the duration of S-phase was observed to be reduced by approximately three hours in the
presence of dXs. This fits well to the reduction of the mitotic delay derived from the mitotic index curves. No influence of dXs on the duration of G + prophase was found at this UV
fluence. It should be emphasized, however, that these measurements of cell-cycle parameters
are only rough estimates since sampling times of 3 hours with colchicine were used to collect
cells in metaphase. In any case, the mitotic indices and the cell-cycle estimates obtained from
the MLI curves indicate that the reduction of the percentage of GCS and of miconucleated
cells by the addition of the four deoxyribonucleosides cannot be explained by enhanced
interphase death.
2
2
2
2
2
2
T A B L E I. Duration o f S-phase and G
U V fluence
+ Prophase as Estimated from M L I Curves*
2
Ojoule/m
S-phase
Post-treatment
0*>
O m M Caffeine,
0 m M dXs
7
-°
G
2
2
2.6joule/m
+ prophase
f t r
(
3
-
)
8
S-phase
<
n
h
r
G
2
5.2joule/m
+ prophase
)
h
(
d
2
n
r
S-phase
>
d
h
(
n
r
G
>
2
+ prophase
(
d
2
n
h
r
>
d
O m M Caffeine,
0.1 m M d X s
6.0
3.8
nd
nd
nd
nd
1.0 m M Caffeine,
0 m M dXs
7.0
3.5
8.0
3.0
9.5
4.1
1.0 m M Caffeine,
0.1 m M d X s
6.5
4.2
6.5
4.2
6.8
4.3
2.0 m M Caffeine,
0 mM dXs
6.3
4.5
6.7
4.5
10.0
5.0
2.0 m M Caffeine,
0.1 m M d X s
7.0
4.5
6.1
4.7
7.0
5.0
*The cell-cycle parameters given were calculated from metaphase labeling index ( M L I ) curves according to the method
of Evans and Scott [ 1 9 6 4 ] .
S-phase (hr): Estimated from the time interval between half o f the m a x i m u m labeling index on the ascending l i m b to
half the m a x i m u m labeling index on the descending limb of the first peak, minus the duration o f the H - t h y m i d i n e treatment (30 minutes).
G + prophase: Estimated from the time interval between the beginning o f H - t h y m i d i n e treatment to the time when
the labeling index reached half o f the m a x i m u m value.
d X s : deoxyadenosine + deoxycytidine + deoxyguanosine + thymidine; concentration 0.1 m M each.
nd = not determined.
F r o m growth curves of exponentially growing cells, a generation time o f 1 3 - 1 4 hours was estimated.
3
3
2
Anticlastogenic Effects of Nucleosides
349
DISCUSSION
1. G C S and Premature Chromosome Condensation
GCS (class E) was the most frequently observed class of altered chromosome
morphology in the present experiments using whole-cell irradiation (X = 254 nm) and
post-treatment with caffeine. Two types of GCS were obtained: 1) Metaphase figures with
chromatid breaks and/or gaps in all chromosomes (Fig. lb); 2) metaphase figures with
"pulverization" [Zorn et al, 1976; Vogel and Bauknecht, 1978] of the chromosomes (Fig.
lc). We have seen many examples of GCS where chromosomes with numerous chromatid
breaks were still present besides pulverized chromosomes. This suggests that types 1 and 2
are closely related to each other. Type 2 was obtained in the large majority of metaphase
figures with GCS. These figures resemble prematurely Condensed chromosomes (PCC)
during S-phase (S-PCC) [Johnson and Rao, 1970; Sperling and Rao, 1974] or Gl-PCC
following UV irradiation [Schor et al, 1975]. Therefore, we have considered the possibility that type 2 figures might be due to micronucleus-derived premature chromosome
condensation [Obe and Beek, 1975]. The temporal relationship between the Observation of shattered chromosomes and the production of micronuclei observed in our experiments, however, rules out such a possibility: Enhanced formation of micronuclei was
observed after mitotic cells with shattered chromosomes appeared in the cultures (compare
Figs. 2, 3, and 5).
Different mechanisms for the induction of GCS and PCC, however, do not exclude
the possibility that pulverized chromosomes in metaphase plates with GCS indicate a
failure of chromosome condensation. Whether the pulverized appearance of chromosomes
in GCS figures is mainly due to a large number of DNA breaks or to a failure of chromosome condensation, remains to be investigated. A model developed by us previously to explain the induction of GCS by UV light and caffeine [Zorn et al, 1977; T. Cremer et al,
1980a, b] is compatible with both possibilities.
2. Antagonistic Effects of Nucleosides
The antimutagenic and anticlastogenic action of deoxyribonucleosides is well known
[Novick and Szilard, 1952; Kihlman, 1977; Gebhart, 1977]. In the present investigation, it
is shown that the synergistic effect of UV light plus caffeine on the induction of GCS and
of micronucleated cells is significantly reduced by the addition of the four dXs to the postirradiation medium. The evaluation of mitotic indices and cell-cycle parameters indicates
that the effect is not due to enhanced interphase death.
A possible explanation of the antagonistic effects of nucleosides on the induction
of chromosome shattering by UV light and caffeine may be obtained by findings of Collins
and Johnson [1979] that the addition of nucleosides may enhance DNA repair synthesis
in UV-irradiated Microtus agrestis cells. Unscheduled DNA synthesis following UV irradiation was observed also in the V79 line used in the present investigation [Zorn, 1978]. This
suggests the following line of reasoning: First, it seems to be generally assumed that the
synergistic action of UV light plus caffeine on the induction of chromosome aberrations
is due to the inhibition of daughter Strand repair (postreplication repair) of DNA photolesions [Nilsson and Lehmann, 1975; Kihlman, 1977; Roberts, 1978]. Here it is assumed
that the inhibition of daughter Strand repair by caffeine plays a decisive role also in the
induction of generalized chromosome shattering: GCS is induced if the number of daughter Strand repair sites exceeds a certain threshold which may vary from cell to cell [T.
350
Cremer, Cremer, and Simickova
Cremer et al, 1980a, b]. Second, addition of nucleosides to the postirradiation medium
may enhance the excision of UV-induced pyrimidine dimers.
If so, the number of remaining DNA photolesions and, hence, the number of
daughter-strand-repair Sites becomes smaller in nucleoside-treated cells than in untreated
ones. If this number falls below the threshold value characteristic for a given cell as first
described, no GCS is induced. Although the model outlined above is far from proven, it
offers a plausible mechanism for the antagonistic effect of nucleosides on GCS and fits
well with the data available. It is also consistent with evidence obtained by Nakano and
co-workers [1979] that the adverse effect of caffeine on the survival of UV-irradiated V79
cells is strongly diminished by an enhancement of the period of time available for excision
repair. While the anticlastogenic effect of nucleosides may be due to their effect on excision-repair capacity, possible effects of an improved supply of cells with nucleosides on
daughter-strand-repair capacity may also be considered.
Caffeine may have an inhibiting effect on the uptake and on the metabolism of DNAprecursors [Lehmann and Kirk-Bell, 1974] (CA. Waldren, personal communication, 1980).
The addition of nucleosides might overcome the adverse effect of a reduced supply of nucleosides. Furthermore, some interference of nucleosides with uptake and/or metabolism
of caffeine, thus reducing the effective intracellular concentration of caffeine, should not
be omitted from consideration.
CONCLUSIONS
Deoxyribonucleosides exert an antagonistic effect on the induction of GCS and micronucleus production by UV light and caffeine in V79 cells. It is suggested that by the addition
of nucleosides, the excision repair capacity may be promoted, thus reducing the number of
caffeine-sensitive Sites of daughter Strand repair (postreplication repair).
ACKNOWLEDGMENTS
This work was supported by grants from the Deutsche Forschungsgemeinschaft
(Wo 148/16 and SFB 46). We thank Dipl Biol Hella Baumann, Brigitte Lechner, and
Ghassan Jabbur for their excellent technical assistance in part of the experiments. We
are greatly indebted to Dr. Beryl Hartley-Asp (Heisingborg, Sweden) for stimulating discussions.
REFERENCES
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Author Index
Abrahamson, Seymour,
1, 4 4 7
Hartman, Philip E., 3
A l l e n , James W., 1 7
H a r t m a n n , A . W . , 187
Anderson, D . , 97
H a s t w e l l , R o w a n M . , 531
A u , William W., 455
Henke, Craig A . , 487
H e n r i c h , R i c h a r d T . , 139
H i n e , C . H . , 59
B a k s h i , K . , 51
Barale, R., 359
H o b b s , C h a r l e s H . , 101
B a r d i n , C . W . , 51
H o l d e n , H e n r y E . , 67
B a u m , J.W., 179
Hooper, A l a n B . , 487
Beck, A . K . , 425
Hopke, Philip K . , 4 1 9
B l o o m , Stephen E . , 435
Hsu, T . C . , 455
Boyce, J.T.,
133
B r u s i c k , D a v i d J . , 5 1 , 3 2 3 , 431
I r w i n , J a m e s , 381
B u l l o c k , L . P . , 51
J a c o b s o n , E l i z a b e t h D . , 389
Carrano, A . V . ,
325
Johnston, Dennis A . , 455
Cheh, Albert M., 487
Johnston, James B., 419
Chrisman, C . L . , 465
C l a r k , C h a r l e s R . , 101
Collie-Bruyere, Cheryl, 455
Kammermeyer,
K . L . , 515
K a p p a u f , H . , 191
C o n n o r , T . H . , 521
Cooper, S.F., 187,
K a i e , P . G . , 179
543
K a r p i n s k y , George E . , 353
K l i g e r m a n , A n d r e w D . , 157
Cremer, C , 339
Krell, Kenneth,
Cremer, T., 339
C r i d e r , P a u l a A . , 67
C r o s s e n , Peter E . , 149
389
L a n g l e y , Charles H . , 167
Legator, M . S . , 43,
L o c k , L . F . , 111,
D a c r e , J a c k C , 531
521
125
L y m a n , R . F . , 133
de B e r t o l d ! , M . , 3 5 9
DeJongh, C , 447
M a c G r e g o r , James T., 405,
Dent, J o h n G . ,
M a t t e r n , L P . , 97
27
Diamond, Martin J., 405
M a z z e n o , L a u r e n c e W., Jr., 4 0 5
M c G r e g o r , D o u g l a s B . , 531
Barle, B . A . , 133
E y a , Bryan K., 395
M c K i n n e l l , Robert G . , 487
Meyer, H . Unger,
447
M e y n e , J u l i a n n e , 4 3 , 521
Friedman, Mendel, 405
M i c h a l o p o u l o s , George,
M i n k l e r , J . L . , 325
Garriott, M . L . , 465
M o l i n a , M . , 371
G a u l d e n , Mary Esther, 473
M o o r e , D . H . , II, 3 2 5
G e b h a r t , E . , 191
M o r g a n , W i l l i a m F . , 149
Giovannetti, M . , 359
Morishima, Akira,
139
G o u l d , Daniel H . , 509
Green, M . H . L . , 97
Nogawa, Takayuki,
Griselli, M . , 359
Guest, Derek,
27
509
P u l l i n , T . G . , 521
549
139
157
550
A u t h o r Index
Preidecker, Barbara Lee Borns,
75,
S t r o m , Stephen C , 157
85
T a l c o t t , R o n a l d E . , 395
Racine, Robert R., 167, 425
T o d d , L o r i A . , 435
R i a c h , C o l i n G . , 531
Traut, H . , 89
R o b e r t s , G l a d w i n T . , 17
T r o l l , W . , 371
R o s e n c r a n z , H e r b e r t S., 3 5 3
R o s s m a n , T . G . , 371
V a l e n c i a , R . , 133
V o e l k e r , R o b e r t A . , 167
S c h w a r t z , Jeffrey L . , 4 7 3
Seiler, J . P . , 97
W a h r e n b ü r g , M a r g i t t a G . , 67
Simickova, M . , 339
W a r d , J . B . , J r . , 521
Skochdopole, Jill, 487
Wehr, C a r o l M . , 509
Soares, E . R . , 3 5 , 1 1 1 , 125
W i l m e r , J a m e s L , , 35
Sonnenblick, B . P . , 201, 547
W o o d r u f f , R . C . , 133
Stetka, D . G . , 325
Würgler, F . E . , 425, 447
S t o l z e n b e r g , S . J . , 59
S t o i i e , D . , 371
Strauss, B e r n a r d , 381
Z i m m e r i n g , S., 1 8 7 , 51 5, 5 4 3
Subject Index
A b s t r a c t s t o the E l e v e n t h A n n u a l
Delta-9-tetrahydrocannabinol,
M e e t i n g o f the E n v i r o n m e n t a l
Development,
Mutagen Society, 230
Diaminotoluene,
A u t h o r index to, 317
1 39
435
111
D i m e t h y l n i t r o s a m i n e , 51
A c t i v a t i o n , 353
Dinitrotoluene,
Aflatoxin B i ( A F - B j ) , 435
DMN,
A i r particulates,
D N A r e p a i r , 7 5 , 3 7 1 , 3 8 1 , 531
75
111
187
A l l o z y m e d e f i c i e n c i e s , 1 67
D o m i n a n t l e t h a l , 111
Anaerobes,
Drosophila
353
females,
Anaphase preparation of
chromosomes,
543
m e l a n o g a s t e r , 8 9 , 1 3 3 , 1 7 9 , 187
139
Dyes, 405
A n e u p l o i d y , 89
Anticlastogens,
191
Antimutagens,
E c o l i , 3 7 1 , 531
339
A r s e n i c , 371
8 - h y d r o x y q u i n o l i n e sulfate,
Aspergillus, 359
E n z y m e i n h i b i t i o n , 27
191
E p o x i d e h y d r a t a s e , 27
B a c t e r i a l m u t a g e n e s i s , 27
Baseline frequency,
E t h y l carbamate, 1 7
325
Benzene, 43
5-Bromodeoxyuridine,
B e n z i m i d a z o l e , 67
Flame retardants, 405
149
B l e o m y c i n , 89
F l u i d i z e d bed c o m b u s t i o n ,
101
Fluorescent lamp, 389
B r o m o d e o x y u r i d i n e , 35, 325
4-(p-nitrobenzyl) Pyridine, 395
F r a c t i o n a t i o n , 75
Caffeine, 339
C a n c e r c h e m o t h e r a p e u t i c drugs, 4 5 5
G a m m a radiation, 473
C a n n a b i d i o l , 139
Gene-conversion,
C a n n a b i n o l , 1 39
Genetic
Cascade impactors,
101
effects,
Cell survival, 473
359
179
f a c t o r s , 51
C h e m i c a l m u t a g e n s , 381
Chick embryo, 435
Halogenated hydrocarbons,
C h i n e s e h a m s t e r cells, 371
Hepatocytes,
Chromosome
H u m a n , 325
aberrations, 473
B lymphocytes,
breakage, 455
l y m p h o c y t e cultures,
loss, 1 8 7 , 5 1 5
T lymphocytes,
shattering,
339
473
139
473
H y d r o x y u r e a , 381
C h r o n i c i r r a d i a t i o n , 167
Ciastogens,
59
157
Hyperthermia, 465
509
G o a l f l y ash, 101
In v i t r o m u t a g e n e s i s assay, 51
C o - c u l t u r e , 157
In v i v o , 1 7 , 35
C y c l o h e x e n e o x i d e , 27
Cyclophosphamide,
Cytogenetics, 43, 465
assays, 4 5 5
lonizing radiation,
157
Lethais, 133
L 5 1 7 8 Y cells, 3 8 9
Screening, 5 0 9
551
179
552
Subject
L o c i number,
Index
Replicative synthesis,
447
chromosomes,
cultures,
381
R F / J m i c e , 51
L y m p h o c y t e , 325
149
191
S a c c h a r o m y c e s cerevisiae, 359,
3 9 5 , 531
Salmonella t y p h i m u r i u m , 353,
M a g n e t i c fields, 179
M a t e r n a l effect,
543
3 9 5 , 4 0 5 , 531
M e d r o x y p r o g e s t e r o n e a c e t a t e , 51
a
mei-9 ,
515
Segregational errors of
Metabolie activation, 435
Metaphase,
m u t a g e n i c i t y test, 101
43
chromosomes,
Mice, 35, I I I , 465
35, 149, 157, 191, 325, 435
Micronucleus, 43, 339, 509
Size-dependent
Mitotic
Spermatogenesis,
arrest, 6 7
MMS,
mutagenicity,
101
465
S p e r m m o r p h o l o g y , 1 1 1 , 125
S p i n d l e d i s r u p t i o n , 67
index, 473
recombination,
139
Sister c h r o m a t i d e x c h a n g e , 1 7,
S p o n t a n e o u s m u t a t i o n rates for
531
lethals and specific loci, 4 4 7
515
Mutagenesis, 59, 3 7 1 , 395
S p o t test, 111
Mutagenicity, 75, 353, 405
Sterility, 133
M u t a t i o n s , 133, 179, 389, 531
S t r u c t u r e - a c t i v i t y r e l a t i o n s h i p , 59
Nitrosocarbamates,
Sunlight, 389
Sun l a m p , 389
395
N-methyl-N'-nitro-Nnitroguanidine,
381
T e s t o s t e r o n e , 51
N O - c a r b a r y l , 395
T e t r y l , 531
Nucleosides, 339
T i s s u e s p e c i f i c , 17
N u l l s , 1 67
T o x i c i t y , 133, 389
Translocations, 89
1,3-dinitrobenzene,
531
1,3,5-trinitrobenzene,
531
Trenimon,
191
T r i c h l o r o p r o p e n e o x i d e , 27
2-acetylaminofluorene
P e l l e t Implantation, 35
(2-AAF),
435
Peripheral blood, 509
2-aminofluorene,
Pesticides, 359
2-nitrofluorene,
353
353
P l a t i n u m , 133
P o t e n t i a t i o n , 51 5
Ultraviolet light, 339
P r o c a r b a z i n e , 51 5
R a d i a t i o n , 125
Rats, 125
Recessive s e x - l i n k e d lethals, 89
Repair deficiency, 1 87, 543
V a p o r phase hydrocarbons,
101
X - l i n k e d recessive l e t h a l s , 4 4 7
X i r r a d i a t i o n , 149