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Transformation
of
Human Epithelial Cells:
Molecular and Oncogenetic

Mechanisms
Editors
George E. Milo, M.S., Ph.D.

Professor, Department of Medical Biochemistry
Director of Carcinogenesis and Molecular Toxicology
Comprehensive Cancer Center
The Ohio State University
Columbus, Ohio
Bruce C. Casto, M.S., Sc.D.

Director of Research
Environmental Health and Research Testing, Inc.
Research Triangle Park, North Carolina
Charles F. Shuler, D.M.D., Ph.D.

Assistant Professor, Center for Craniofacial Molecular Biology
University of Southern California
Los Angeles, California
CRC
\CP* J Taylor & Francis Group
^***^/ Boca Raton London New York
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First published 1992 by CRC Press
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PREFACE
Several years ago, in the 1960s and 1970s, there were but a few human
cell lines available to study human cell carcinogenesis. At that time the
"Hayflick" hypothesis suggested that human cells cultured in vitro have a
finite life-span of approximately 60 PDs; little was known of the "real"
relationship between the limited life-span of the human fibroblast in vitro and
aging in vivo. This area of research piqued the interest of the scientific
community because the expression of cancer was thought to be an escape of
the neoplastic phenotype from the limited proliferative potential, i.e., the
finite life-span. However, over the years many human cancers were deter-
mined to be of an epithelial origin, and it was not possible to isolate, except
on a rare occurrence, stable tumor phenotypes in vitro that exhibited infinite
life-spans. Consistently heterogenous human tumor phenotypes either ceased
to proliferate or terminally differentiated in vitro. Rarely did we observe in
a routine fashion an escape from a limited life-span to a phenotype with an
unrestricted proliferative potential. With the advent of collagen-coated plastic
substratum, feeder layers, "quasi"-chemically defined growth media con-
taining the likes of pituitary extracts, and growth factors, epithelial cells from
different human tissues could be isolated and cultured for limited defined
periods in vitro. Still the concept of the limited life-span of normal cells in
vitro persisted and the cancer cell was thought to be an escape from a limited
life-span. This loss of growth control and extension of life-span are discussed
in Chapter 5 by Johng S. Rhim, one of the pioneers in the field of SV-40T-
induced immortalization of human cells. We have included also a chapter on
another developing field that will dramatically impact the field of growth
control and life-span. Chapter 2 examines how autocrine and paracrine factors
elicit proliferative growth responses in normal and transformed cells. In the
1970s, Heppner recognized along with others that epithelial tumors were
heterogenous in cellular composition. Since that time we have learned how
to identify different phenotypes from the epithelial tumors. Because of the
recent achievements by Bert Vogelstein in the late 1980s of identifying, in
colon carcinoma tumors, cells that occupy different stages of progression and
the discovery by Patricia Steggs and Lance Liotta in the late 1980s of the
molecular events that were associated with expression of the metastatic stage,
we can now follow the events of metabolism of putative human carcinogens,
DNA-adduct binding, early, middle and late stages of progression of initiated
cells, anchorage-independent growth, identification of expression of altered
phenotypes, and identification of expression of tumorigenic and metastatic
phenotypes. If we agree that tumors are clonal in origin, then we need to
understand that tumor heterogeneity may be a function of the composition of
mixed phenotypes. Some of these questions comprise the reasons this book
and the contributors were assembled to address the stages from metabolism
to metastasis of epithelial cells associated with epithelial tumors.
In Chapter 1, James R. Smith addresses the role immortalization plays
in cancer and how resistant human cells are to spontaneous immortalization.
There has been an attempt to link the loss of expression of a finite life-span
to the change in genetic program of either the 1st or the 4th chromosome. A
defect in these chromosomes putatively can give rise to the expression of a
cellular phenotype that exhibits immortalization. However, at present (in these
authors' opinion) the question of quiescence vs. senescence in human cells
has not been answered by these experiments. The loss of senescent control
can lead to immortalization; the loss of control of expression of quiescence
cannot. However, it is recognized that the transient down-regulation of the
cyclin gene or the permanent interruption of this gene function may play a
pivotal role in the two processes. The next several chapters discuss the stages
of multi-stage carcinogenesis in different epithelial cell systems. Moreover,
correlative stages of progression in different carcinogen-transformed epithelial
cell systems are compared and contrasted. The last few chapters — 10, 11,
and 12 — discuss molecular controlling mechanisms that are involved in the
control of expression of stages of progression, e.g., the role of oncogenes
and their interactiveness with the suppressor genes. In particular, the role of
ras gene mutations and suppressor gene interaction with these activated on-
cogenes in tumorigenic cells in controlling the progression of initiated cells
into tumor cells is presented. The promotion stage in human cell carcino-
genesis is a silent stage and has not been observed experimentally, and many
attempts to discover this stage have led to failure.
In Chapter 13 we compare and contrast the stages from metabolism, DNA-
adduct formation, anchorage-independent tumor growth, and regression be-
tween epithelial cells and fibroblast (see also Transformation of Human Dip-
loid Fibroblast: Molecular and Genetic Mechanisms, Milo, G. E. and Casto,
B . C . , Eds., CRC Press, 1990). Lastly, an invitation was extended to Juergen
R. Vielkind to contribute a guest chapter discussion on the nature of growth
control in osteichthyes. Our reasons for including this chapter are to develop
and understand the biological significances of the conservation of suppressor
gene function phylogenetically from lower animals up to humans and to
understand the role of some of these genes in more primitive animal systems.
It was our intention to present to the scientific community a forum for
focusing on the utility of how human epithelial cells as systems can be used
to examine how environmental xenobiotics can be metabolized to reactive
metabolites that can react with critical sites on the genomic DNA that lead
to the expression of an early stage of a transformed phenotype. Later in the
book we have focused on how current dogma in suppressor gene-oncogene
interaction is insufficient to totally explain the existence of many diverse
phenotypes in a heterogenous spontaneous epithelial cell tumor. Moreover,
the concept of plasticity — the transient expression of a tumorigenic phen-
otype — is not explained only by the presence of mutations in critical sites
of activated oncogenes or mutations in suppressor genes. This treatise is an
attempt to collate many of the scientific results, significant scientific concepts,
and laboratory efforts from active investigators in this field of environmentally
induced human epithelial cell cancer.
George E. Milo
Bruce C. Casto
Charles F. Shuler
THE EDITORS
George E. Milo, B.A., M.S., Ph.D., is Professor of Medical Biochem-

istry in the College of Medicine and Program Director of Molecular Envi-
ronmental Health in the Comprehensive Cancer Center at The Ohio State
University in Columbus.
Dr. Milo has actively pursued research in the discipline of human cell
carcinogenesis since 1970. He published the first report on the transformation
of human fibroblast cells in 1978 in Nature and the first report on the trans-
formation of human epithelial cells in Cancer Research in 1981. Again in
1990, along with Dr. Charles Shuler, he published a new concept on the
isolation and identification of a plastic anchorage-independent growth of a
nontumorigenic phenotype that can be transiently converted to a tumorigenic
and metastatic phenotype. This article was published in the Proceedings of
the National Academy of Science (U.S.A.). His National Institutes of Health
(NIH)-supported postdoctoral training at the Roswell Park Memorial Institute
in Cancer Research (Buffalo, NY) served him well as a stepping stone to
begin his career in human cell carcinogenesis.
Dr. Milo is a member of the Society for Toxicology — Molecular Tox-
icology Division, the American Association for Cancer Research, the Amer-
ican Society for Biochemistry and Molecular Biology, the American Society
for Cell Biology, and the International Society for the Study of Xenobiotics.
He has published in excess of 130 publications in the discipline of human
cell carcinogenesis. He has written several chapters for different books on
the subject and has served as an ad hoc reviewer for the National Institutes
of Health — National Cancer Institute (NIH-NCI) in the discipline, as a
reviewer and Chairperson for the U.S. Environmental Protection Agency
Extramural Health Effects Research Review Panel, as a Chairperson for the
U.S. Environmental Protection Agency ''Health" Research Centers program,
and as a reviewer on the NIH-NCI Parent Preclinical Pharmacology Program
Panel. He has received many grants from the National Institutes of Health
— National Cancer Institute and National Institute of Environmental Health
Science — and from the U.S. Environmental Protection Agency — Health
Effects Research. He also edited, along with Dr. Bruce C. Casto, Transfor-
mation of Human Fibroblasts: Molecular and Genetic Mechanisms, published
by CRC Press in 1990.
His present area of interest is to investigate how exposure to environmental
xenobiotics alters human gene function.
Bruce C. Casto, M.S., Sc.D., is Director of Research for Environmental

Health and Research Testing, Inc. in Research Triangle Park, North Carolina.
Dr. Casto received training in microbiology and virology at The Ohio State
University and the University of Pittsburgh. During this time, research was
conducted in the areas of viral receptors, oncolytic viruses, and adeno-as-
sociated viruses. While at the University of Pittsburgh, Dr. Casto discovered
the defective nature of AAV-1 and its dependence on adenovirus for repli-
cation. He was an assistant member at the Institute for Biomedical Research
— American Medical Association, professor of microbiology at Rush Medical
School, senior scientist at BioLabs, Inc., and research director for health
effects at Northrop Environmental Sciences. Dr. Casto's major area of re-
search is chemical carcinogenesis, especially the enhancement of viral trans-
formation by chemical carcinogens and the chemical transformation of mam-
malian cells in vitro.
Charles F. Shuler, D.M.D., Ph.D., is Assistant Professor in the Center

for Craniofacial Molecular Biology at the University of Southern California
School of Dentistry.
Dr. Shuler received his dental degree from the Harvard University School
of Dental Medicine, his Oral Pathology training at the University of Min-
nesota, and his Ph.D. in Experimental Pathology from the University of
Chicago.
Dr. Shuler is a member of the American Society for Cell Biology, the
American Association for Dental Research, the American Association for
Advancement of Science, and the American Academy of Oral Pathology. He
has served as an Associate Editor of the Journal of Oral Pathology.
His current areas of research interest include mechanisms of epithelial
differentiation, especially during the development of the secondary palate in
utero, and human cell transformation and tumorigenesis.
CONTRIBUTORS
William M. Baird, Ph.D. Frank C. Cuttitta, Ph.D.

Glenn L. Jenkins Professor of Biomarker and Prevention
Medicinal Chemistry Research Branch
and Purdue Cancer Center NCI-Navy Medical Oncology
Purdue University Branch
West Lafayette, Indiana Biotherapy Section
National Naval Medical Center
Linda L. Barrett, M.S. National Cancer Institute
School of Medicine and Department of Medicine
East Carolina University Uniformed Services University of
Greenville, North Carolina the Health Sciences
Bethesda, Maryland
Michael J. Birrer, M.D., Ph.D.
NCI-Navy Medical Oncology
Curtis C. Harris, M.D.
Branch
Laboratory of Human
Clinical Oncology Program
Carcinogenesis
Division of Cancer Treatment
National Cancer Institute
National Cancer Institute
Bethesda, Maryland
Bethesda, Maryland
David G. Kaufman, M.D.,
Tammela Butler, B.S.
Ph.D.
Department of Toxicology
Department of Pathology
University of North Carolina
University of North Carolina
Chapel Hill, North Carolina
School of Medicine
Charleata A. Carter, Ph.D.
Experimental Carcinogenesis and
Mutagenesis Branch Hudson H. S. Lau, Ph.D.
National Institutes of Department of Medicinal
Environmental Health Services Chemistry Pharmacognosy
Research Triangle Park, North Purdue University
Carolina West Lafayette, Indiana
Bruce C. Casto, M.S., Sc.D.

Caroline H. Laundon, Ph.D.
Environmental Health & Research
GeneCare
Testing, Inc.
Research Triangle Park, North
Carolina
Teresa A. Lehman, Ph.D.
Dharam P. Chopra, Ph.D. Laboratory of Human
Institute of Chemical Toxicology Carcinogenesis
Wayne State University National Cancer Institute
Detroit, Michigan Bethesda, Maryland
George E. Milo, Ph.D. James R. Smith, Ph.D.
Department of Medical Roy M. & Phyllis Gough
Biochemistry Huffington Center on Aging
and Department of Molecular and Division of Molecular
Environmental Health of the Virology
Comprehensive Cancer Center Baylor College of Medicine
The Ohio State University Houston, Texas
Columbus, Ohio
Martha R. Stampfer, Ph.D.
Zenya Naito, M.D., Ph.D. Department of Cell and Molecular
Medical Technology Biology
Yokosuka National Hospital Lawrence Berkeley Laboratory
Yokosuka, Kanagawa, Japan University of California
Berkeley, California
Johng Sik Rhim, M.D.
Department of Radiation Medicine Gary M. Stoner, Ph.D.
Georgetown University School of Experimental Pathology
Medicine Department of Pathology
Washington, D.C. Medical College of Ohio
and Laboratory of Cellular and Toledo, Ohio
Molecular Biology
National Cancer Institute Juergen R. Vielkind, Ph.D.
Bethesda, Maryland Department of Cancer
Endocrinology
Clifford A. Rinehart, Ph.D. British Columbia Cancer Agency
Department of Pathology and Department of Pathology
University of North Carolina University of British Columbia
Chapel Hill, North Carolina Vancouver, British Columbia,
Canada
Charles F. Shuler, D.M.D.,
Ph.D. Li Hui Xu, M.D.
Center for Craniofacial Molecular Department of Pathology
Biology University of North Carolina
School of Dentistry Chapel Hill, North Carolina
University of Southern California
Los Angeles, California Paul Yaswen
Department of Cell and Molecular
Jill Siegfried, Ph.D. Biology
Department of Pharmacology Lawrence Berkeley Laboratory
University of Pittsburgh University of California
Pittsburgh, Pennsylvania Berkeley, California
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TABLE OF CONTENTS
Chapter 1
In Vitro Cellular Aging and Immortalization 1
James R. Smith
Chapter 2
Detection of Growth Factor Effects and Expression in Normal and
Neoplastic Human Bronchial Epithelial Cells 13
Jill M. Siegfried, Michael J. Birrer, and Frank C. Cuttitta
Chapter 3
Human Cell Metabolism and DNA Adduction of Polycyclic Aromatic
Hydrocarbons 31
Hudson H. S. Lau and William M. Baird
Chapter 4
Human Esophageal Epithelial Cells: Immortalization and In Vitro
Transformation 67
Gary D. Stoner, Zenya Naito, and George E. Milo
Chapter 5
Transformation of Human Endometrial Stromal Cells In Vitro 85
Clifford A. Rinehart, Charleata A. Carter, Li Hui Xu, Linda L. Barrett,
Tammela Butler, Caroline H. Laundon, and David G. Kaufman
Chapter 6
Factors Influencing Growth and Differentiation of Normal and
Transformed Human Mammary Epithelial Cells in Culture 117
Martha R. Stampfer and Paul Yaswen
Chapter 7
Transformation of Colon Epithelial Cells 141
Dharam P. Chopra
Chapter 8
Multistep Carcinogenesis and Human Epithelial Cells 169
Johng S. Rhim
Chapter 9
Morphologic and Molecular Characterizations of Plastic Tumor Cell
Phenotypes 211
Charles F. Shuler and George E. Milo
Chapter 10
Oncogene and Tumor Suppressor Gene Involvement in Human Lung
Carcinogenesis 235
Teresa S. Lehman and Curtis C. Harris
Chapter 11
Events of Tumor Progression Associated with Carcinogen Treatment
of Epithelial and Fibroblast Compared with Mutagenic Events 261
George E. Milo and Bruce C. Casto
Chapter 12
Progression from Pigment Cell Patterns to Melanomas in Platyfish-
Swordtail Hybrids — Multiple Genetic Changes and a Theme for
Tumorigenesis 285
Juergen R. Vielkind
Index 303
Smith 1
Chapter 1
IN VITRO CELLULAR AGING AND IMMORTALIZATION
James R. Smith
TABLE OF CONTENTS
I. Introduction 2
II. In Vitro Cellular Aging is Dominant in Somatic Cell Hybrids 3

A. Limited In Vitro Life-Span of Normal Cells 3
B. Hybrids between Normal and Immortal Cells 3
C. Fusion of Immortal Cells with Immortal Cell Lines 4
D. Microcell Hybrid Experiments 6
III. Cellular Aging Is an Active Process 6

A. Heterokaryon Experiments 6
B. Membrane-Associated DNA Synthesis Inhibitors of
Senescent Cells 7
C. Inhibition of DNA Synthesis by Poly (A) + RNA 8
IV. Discussion 9
References 10
2 Transformation of Human Epithelial Cells
I. INTRODUCTION
Carcinogenesis has become widely accepted as a multistep process2 (see

References 1 and 2 for recent reviews). A number of events have to occur
in order for cells to become cancerous. In many cases, if not all, cellular
immortalization is one of these steps and is an obligatory process.3 Normal
human diploid cells go through various numbers of population doublings
(PDs), depending on the age of the donor and the origin of the tissue from
which the cells are derived.4"6 In most cases, cells from adult donors go
through fewer PDs than cells from young or embryonic donors.6 The number
of PDs that a culture can go through when derived from adult tissue is typically
20 to 30.5 Cancers generally are of clonal origin, and for a single cell to
produce a tumor 1 g in size requires approximately 30 cell divisions. Primary
tumors are not the major cause of problems in carcinogenesis because of the
possibility of surgical removal of the primary tumor and, hence, the threat
to the individual by that tumor. Indeed, metastasis is the crucial step that
causes carcinogenesis to be a life-threatening phenomenon. Metastases are
also generally of clonal origin and require a cell that has already gone through
a number of doublings in the primary tumor to undergo further doublings as
a metastatic growth in order to be significant. Cell growth, tumor regression,
and cell death are all normal parts of the processes of carcinogenesis. There-
fore, the number of PDs that cells have to go through in order to become
life-threatening may be more than 100 to 200. This range is clearly greater
than normal cells are able to go through, as evidenced by experiences with
human fibroblasts in tissue culture. 7 Other cells in the body may normally be
able to go through more doublings in vivo. However, at this time, the doubling
limit for most epithelial cells in situ is unknown. Therefore, it is reasonable
to assume that as part of the multistep process of carcinogenesis, cellular
immortalization is required for tumor progression and metastasis. The spon-
taneous immortalization of human cells in culture has never been observed.
However, this can be contrasted with the situation that we see in rodent cells,
particularly mouse and rat cells, in which spontaneous immortalization is the
rule rather than the exception. 810 When considering whether immortalization
may be necessary for tumor formation and metastasis, it is interesting to
compare the rates of tumor formation in rodents with those in humans. A
mouse weighs on the order of 10 g while humans weigh on the order of 100 kg,
and the mouse's life-span is approximately 1/30 that of a human, yet mice
very often have tumors during their 2V2- to 3-year life-span. Therefore, on
a per cell unit time basis, the rate of tumor formation in mice is 105 to 106
times the rate of tumor formation in humans. It seems likely that this incredibly
higher rate of tumor formation seen in mice compared to humans is due to
the much higher incidence of spontaneous immortalization of mouse cells
compared with human cells. Therefore, the study of cellular immortalization
and of mechanisms that limit the proliferative potential of normal human cells
in culture is of paramount importance in understanding the mechanisms of
carcinogenesis in humans.
Smith
II. IN VITRO CELLULAR AGING IS DOMINANT IN

SOMATIC CELL HYBRIDS
A. LIMITED IN VITRO LIFE-SPAN OF NORMAL CELLS

Swim and Parker were the first to show that human fibroblasts derived
from biopsies had a limited proliferative potential in culture. 11 Hay flick and
Moorehead in 1961 showed that these cells were karyotypically normal and
that normal cells derived from a large number of different individuals all had
a finite proliferative potential.4 They also showed that a major characteristic
of cells that were able to divide indefinitely, i.e., transformed immortal cells,
was an abnormal karyotype. In 1965, Hayflick proposed that the limited in
vitro proliferative potential of normal human fibroblasts in culture was a
manifestation of aging at the cellular level.5 More recently, it has been pro-
posed by O'Brien et al.3 that limited proliferative potential of normal cells
in vitro and also in vivo is a powerful tumor suppressor mechanism. The
observation of limited proliferative potential of normal cells in culture has
been repeated in hundreds of labs and thousands of cultures over the last 30
years.7 The proliferative potential of the cells depends on the age of the
donor,6 species of the donor,12 and the site of biopsy.6 Typically, human
embryonic cells will undergo 50 to 80 PDs before growth cessation, although
it has been reported that some cells are capable of going through approximately
100 PDs before proliferation stops.13 Cells from other species go through
fewer PDs than those from humans, the exception being the Galapagos turtle.14
The number of PDs that the cells are able to undergo is correlated with the
maximum life-span of the species.
Cells spontaneously immortalize at various rates, depending on the species
of origin of the cells. Human cells and chick cells have never been observed
to immortalize spontaneously, while rodent cells routinely immortalize in
culture and species such as bovine immortalize spontaneously only rarely.15
The mechanisms that lead to limited in vitro proliferative potential of normal
cells in culture is not understood. A number of investigations have been carried
out over the past 30 years to measure various biochemical, metabolic, and
structural parameters of these cells as they age in culture, and with very few
exceptions, which will be discussed later, no changes have been observed
that could account for the irreversible division cessation. Likewise, the process
by which cells escape the finite proliferative potential and become able to
divide without limit (immortalization) is not understood. In order to try to
understand the mechanisms operating in these processes, we and others have
undertaken a series of experiments discussed below.
B. HYBRIDS BETWEEN NORMAL AND IMMORTAL CELLS

The early work of Littlefield suggested that the limited proliferative po-
tential (the senescence phenotype) might be dominant in somatic cell hybrids
between senescent cells and young proliferating cells.16 However, the evi-
dence was not conclusive and the prevailing belief at that time was that cellular
immortalization was due to dominant changes in the cellular genome. Many

different ideas have been presented to try to explain the limited proliferation
of normal cells in culture. These can be broken into two main categories.
One category proposes that cells stop dividing because they accumulate dam-
age of various sorts, e.g., somatic cell mutations or errors in protein synthesis,
so that the error burden becomes so large that the cells are no longer able to
divide. The other category proposes some sort of genetic program that limits
the in vitro life-span of cells in culture. We thought that we might be able
to differentiate between these two broad categories of hypothesis by fusing
normal cells with immortal cells and determining whether the hybrids resulting
from that fusion had a limited in vitro life-span or were immortal. If normal
cells stopped dividing because they had accumulated a large amount of dam-
age, then one could argue that cells that are immortal have escaped from
limited proliferative potential because either they don't accumulate damage
at the same rate or they have evolved a mechanism to better cope with the
damage. Therefore, one might expect in hybrids that the phenotype of cellular
immortality would be dominant.
In the first set of fusions, we fused an immortal SV40-transformed cell
line with a normal cell line that was at the end of its in vitro life-span.17 We
observed that the hybrid colonies proliferated for various numbers of PDs and
then stopped dividing. About 70% of the colonies were able to go through
fewer than 8 PDs, while the other 30% were able to go through a range of
PDs varying from 30 to 60, but they all stopped dividing. We also showed
that all of the clones expressed the S V40 large T-antigen which is thought to
be the immortalizing agent for normal human diploid fibroblasts infected with
SV40 virus. In order to investigate the generality of this phenomenon, we
fused a number of different cell lines with normal human diploid fibroblasts
and observed the same results in all cases.18 The hybrids had finite proliferative
potential. In all the fusion experiments, we found that immortal variants arose
in the culture at a frequency of approximately 1 per 105 to 106 cells. This is
a much greater frequency of immortalization than that observed in normal
diploid fibroblasts. The tentative explanation for this is that in hybrids, chro-
mosomal segregation takes place and the hybrids lose a chromosome which
encodes a gene that causes the finite proliferative potential. Conclusions from
these experiments are that the limited life-span of normal cells in culture is
dominant over the phenotype of cellular immortality and that cells become
immortal because they lose some of the program that is necessary to impose
a limited proliferative potential on normal cells in culture.
C. FUSION OF IMMORTAL CELLS WITH OTHER IMMORTAL

CELL LINES
If cellular immortality results from recessive changes in the cellular ge-
nome, then we might expect that different defects could occur to render a
cell immortal. If that is the case, then fusion of cell lines having one defect
with cell lines having another defect could result in complementation, giving
Smith
a hybrid that has finite proliferative potential. On the other hand, fusion of
cell lines having the same defect would not result in complementation and
would give rise to hybrids that could divide indefinitely. Therefore, we would
predict that hybrids resulting from fusions among different immortal cell lines
would give two different kinds of results. In one case, some hybrids would
have a finite life-span and the other hybrids would have an indefinite life-
span. In a series of experiments, Pereira-Smith and Smith fused different cell
lines with each other and observed the proliferative phenotype (either finite
or indefinite),19 and assigned more than 30 different cell lines to four different
complementation groups. In order to begin the process of complementation
group assignment, one SV40-transformed cell line was chosen at random to
be representative of complementation group A. Other cell lines were fused
with it. Those that had an indefinite proliferative potential also assigned to
complementation group A; those that had a finite proliferative potential as-
signed to a different complementation group. Using HeLa as a prototypic cell
line for complementation group B, we repeated the process and assigned cell
lines to complementation group B. Cell lines were assigned to other com-
plementation groups in a similar fashion. This process involved numerous
cell fusions, and in no case did we find a cell line that assigned to more than
one complementation group. This indicated that the processes resulting in
cellular immortality were very rare, with no cell lines carrying two different
defects. We looked at a large number of different cell lines resulting from
different kinds of tumors, different cell types of origin, cell lines derived
from different embryonic layers, and cell lines that contained activated on-
cogenes, and in no cases did these parameters affect complementation group
assignment. The only parameter that did affect assignment was immortali-
zation by the SV40 large T-antigen. Seven out of eight of the SV40-immor-
talized cell lines assigned to complementation group A. One of the SV40 cell
lines failed to assign to complementation group A. The reason for this is not
known. We can speculate that the SV40 T-antigen was not the actual im-
mortalizing agent in this case, but was only coincidental in the transformation
process. The assignment of cell lines to different complementation groups
allows us to take a systematic approach to trying to understand what processes
might have occurred to result in cellular immortalization. We speculate at the
present time that those cell lines which assign to the same complementation
group have become immortalized by the same genetic defect. The case for
this interpretation is strengthened by the fact that almost all of the SV40-
transformed immortalized cell lines fall into the same complementation group.
However, it appears that not all DNA tumor viruses immortalize cells by the
same mechanism, because we found that cell lines immortalized by adeno,
papilloma, and herpes virus fell into different complementation groups. Efforts
are currently underway to find the genetic defect that leads to immortalization
in the case of SV40 T-antigen.
D. MICROCELL HYBRID EXPERIMENTS

The introduction of single normal human chromosomes into immortal
cell lines represents a considerable refinement over the techniques of somatic
cell hybridization involving whole cells discussed above. The use of microcell
hybrid techniques has allowed us to assign genes coding for normal cellular
aging processes to one particular human chromosome.
Ning et al.20'21 introduced chromosome 11 from a normal human cell line
into immortal cell lines representative of all four complementation groups.
They observed no effect on growth rate or the immortal phenotype of these
cells. There was some minor and variable effect on tumorigenicity when cells
carrying the intact human chromosome 11 were injected into nude mice.
There was, in some cases, suppression of tumorigenicity and, in other cases,
a delay in the formation of tumors.
Ning et al.22 further showed that introduction of a normal human chro-
mosome 4 into cell lines assigned to complementation group B restored the
phenotype of limited proliferative potential. However, when the human chro-
mosome 4 was introduced into cell lines assigning to the other complemen-
tation groups (A, C, and D), there was no decrease in proliferation potential
and the phenotype of immortality was retained. Thus, it seems clear that genes
on chromosome 4 code for some part of the genetic program that limits the
division potential of normal cells in culture. Disruption of these genes leads
to cells with an immortal phenotype. Sugawara et al.23 found a similar result
in studies in which they introduced the normal human chromosome 1 into
Chinese hamster cells. Human chromosome 1 was able to restore the cellular
aging phenotype in these immortal hamster cells. It remains to be seen whether
chromosome 1 plays a role in the immortalization of human cells.
III. CELLULAR AGING IS AN ACTIVE PROCESS
A. HETEROKARYON EXPERIMENTS
One of the first experiments that gave us an idea of the kinds of processes
that might be responsible for cellular senescence was performed by Norwood
et al.24 and independently by Stein and Yanishevsky. 25 They fused senescent
cells that had reached the end of their in vitro life-span with normal cells that
were still able to proliferate and asked whether the nuclei contained in the
heterokaryon were able to synthesize DNA. When senescent cells were fused
with young cells, it appeared that the senescent cell was able to suppress the
initiation of DNA synthesis in the young cell nucleus, i.e., neither the young
cell nucleus nor the senescent cell nucleus synthesized DNA in the hetero-
karyons up to 72 h after fusion. However, if young cells were fused with
each other, there was no decrease in the ability of the young cell nuclei to
synthesize DNA in the homodikaryon. From these results, it was concluded
that senescent cells produce an inhibitor of DNA synthesis which is able to
act in trans to inhibit the initiation of DNA synthesis in the young nucleus.
Furthermore, it has been shown that senescent cells, when fused with various
Smith
immortal cell lines,25 suppress DNA synthesis in the nucleus of the immortal
cell. This indicates that the inhibitor produced by senescent cells is able to
also inhibit the initiation of DNA synthesis in certain immortal cell lines.
However, other immortal cell lines,26 in particular those that have been im-
mortalized by DNA tumor viruses, e.g., SV40-transformed cells of HeLa
cells (HeLa is known now to have part of the herpes virus DNA integrated
into its genome), are able to induce DNA synthesis, in the short term, in the
senescent cell nucleus. This indicates that although senescent cells produce
an inhibitor of DNA synthesis, it is possible, through the intervention of DNA
tumor viruses, to temporarily override this inhibitor.27
Yanishevsky and Stein showed that the inhibitor of DNA synthesis present
in senescent cells could not interrupt ongoing DNA synthesis, but acted to
block the initiation of DNA synthesis. They found that if senescent cells were
fused to young cells that were more than 3 or 4 h from the S-phase, then
initiation of DNA synthesis was blocked. If they were closer to the S-phase,
then initiation of DNA synthesis was not blocked.28
B. MEMBRANE-ASSOCIATED DNA SYNTHESIS INHIBITORS OF

SENESCENT CELLS
Although the production of an inhibitor by senescent cells is a simple
and attractive explanation for the above results, there may be other expla-
nations. For example, if senescent nuclei were depleted of some factors that
were needed for induction of DNA synthesis and competed with the young
cell nucleus for those factors, then the concentration of positive regulatory
factors could fall below a critical threshold in the heterokaryons. In order
to rule out that possibility, we initiated experiments in which we prepared
enucleated cytoplasms from senescent cells and fused them to whole young
cells, and then asked whether the senescent cytoplasts could cause inhibition
of the initiation of DNA synthesis in the resulting cybrids. We found that
senescent cytoplasts were indeed capable of inhibiting the initiation of DNA
synthesis in young-cell cybrids.29"31 There was an approximately 50% decrease
in the number of young-cell nuclei synthesizing DNA in the senescent-young
cell cybrids compared to cybrids from young-cell cytoplasts fused with young
whole cells. We next asked the location of the inhibitor of DNA synthesis in
senescent cytoplasts. By treating the senescent cytoplasts with trypsin under
conditions that would limit the penetration of the trypsin into the cell and
limit intracellular damage by trypsin (4°C for 1 min), we were able to show
that the inhibitory activity resided on the outside surface of the membrane.
Further evidence for this conclusion was obtained by preparing membrane-
enriched fractions from senescent cells and adding them to young-cell cultures.
These membrane-enriched fractions were very effective in inhibiting the ini-
tiation of DNA synthesis in young-cell cultures.31-32 Furthermore, proteins
extracted from the membrane preparations and added directly to cultures of
young cells were also effective in inhibiting the initiation of DNA synthesis.31
We next examined the role of protein synthesis in the production of this
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inhibitor by treating the cells with cyclohexamine or puromycin at concen-

trations which would inhibit protein synthesis by at least 90% . We found that
a relatively short treatment, approximately 2 h, with cyclohexamine or pu-
romycin was sufficient to eliminate the inhibitory activity from senescent
cells. Upon removal of cyclohexamine from the culture and incubation of the
cytoplasts in the absence of cyclohexamine, inhibitory activity was regained
in about 4 h,31 indicating that cytoplasts were still active and able to synthesize
DNA. Further, this indicated that the messenger RNA coding for the inhibitor
was relatively long-lived.
C. INHIBITION OF DNA SYNTHESIS BY POLY(A)+

In order to explore the feasibility of searching for cDNA clones coding
for the inhibitor, we microinjected poly(A)^ RNA from senescent cells into
young proliferation-competent cells to determine whether inhibitory activity
could be conferred by the messenger RNA. We found that it was strongly
inhibitory.33 By microinjecting different amounts of RNA into the cells, we
were able to calculate that the inhibitor RNA was present in relatively large
abundance (0. 1 to 1 % of the total messenger RNA consisted of inhibitor RNA
according to our calculations).
We also studied the possibility that nongrowing tissues, e.g., rat liver,
might produce an inhibitor. Rat liver was chosen because it can exist either
in a state of nonproliferation or in a state of proliferation. Lumpkin et al.34
found that nonregenerating liver RNA was able to block the initiation of DNA
synthesis when microinjected into young human fibroblasts, whereas the RNA
isolated from regenerating rat liver had no inhibitory activity; indeed, it had
a stimulatory activity. This raised the possibility of using rat liver as a source
of RNA to carry out syntheses and screening of a cDNA library to isolate
genes that were expressed in nonregenerating liver but not expressed in re-
generating liver. Inhibitory RNA has been hybrid selected by cDNA clones
isolated by differential screening of a cDNA library made from nonregener-
ating rat liver poly(A) + RNA. 35 However, these clones do not code for a
messenger RNA that is upregulated in senescent cells (unpublished data).
Other investigators have confirmed these results using RNA from human
liver36 and have extended them to show that RNA isolated from resting human
T-lymphocytes also will inhibit initiation of DNA synthesis when injected
into proliferation-competent human fibroblast cells.37 The results from these
microinjection experiments indicate that it would be feasible to search for
cDNAs coding for the inhibitory messenger RNA and the inhibitory protein.
Construction of cDNA libraries and screening by differential screening using
probes made from young cells and senescent cells is now underway in several
laboratories. Using this approach, various cDNA clones have been isolated;
however, as of yet, no clones have been isolated that have been shown to
code for the inhibitory activity expressed in senescent cells. Although the
cDNA cloning of inhibitor genes has not been fruitful to date, investigators
are still optimistic that straightforward ( + )/( — ) screening of cDNA libraries
Smith
produced from senescent cells or cells from premature aging syndrome patients
will yield the inhibitor genes that are associated with cellular senescence.
Another approach to this problem is to produce monoclonal antibodies
by immunizing mice with surface membrane preparations from senescent
cells. We have isolated monoclonal antibodies in this way.38 Of approximately
6000 hybridoma cultures screened, three yielded antibodies that reacted pref-
erentially with senescent cells but not with young cells. All the antibodies
react with an epitope on the fibronectin molecule. Although they react with
fibronectin from various sources, when it is denatured, they only react with
fibronectin distributed on the surface of senescent cells, not with fibronectin
distributed on the surface of young cells. This indicates that senescent cells
are producing either a fibronectin that is altered in its primary structure or
posttranslationally modified in a way different from that of young cells. The
senescent cell-produced fibronectin would then have a different conformation
than that of young cells. Another possibility is that the fibronectin protein
produced by senescent cells and young cells is the same, but the interaction
of fibronectin with other molecules produced by senescent cells and young
cells is different, thus leading to an altered confirmation of fibronectin as-
sociated with senescent cells, exposing an epitope which is sequestered in
young cells.
IV. DISCUSSION
The experimental results reviewed here indicate that cellular aging is an

active process, perhaps part of a genetic program, and that this genetic program
can be disrupted in various ways, giving rise to cellular immortality. We have
shown that there are at least four different ways that the normal cell processes
can be disrupted to lead to cellular immortality. One of the processes that
takes place in normal cells seems to be production of an inhibitor of the
initiation of DNA synthesis. This inhibitor is produced in quiescent cells, but
is reversible by the addition of growth factors or serum mitogens. The inhibitor
produced by senescent cells is not reversible. We do not know at the present
time whether the inhibitor produced by quiescent cells is the same as the
inhibitor produced by senescent cells. It is possible that they are the same,
but the control of expression of the inhibitors is different between young cells
and senescent cells. In senescent cells, the inhibitor is being produced con-
stitutively, whereas in young cells it is modulated in different parts of the
cell cycle and by growth factors.
Recently, changes in a number of cell cycle genes as cells become se-
nescent have been reported. These include failure to express c-fos or cdc2
when the cells are mitogenically stimulated,39'40 and the failure to phospho-
rylate the Rb gene.41 It may be that the defects in the regulation of these cell
cycle-related genes are responsible for senescent cells being unable to enter
into the S-phase. However, according to the data presented above, these
changes would be secondary to the expression of a cell surface inhibitor of
DNA synthesis by senescent cells. The mechanism by which this inhibitor

changes the expression of cell cycle genes remains to be elucidated. One
possibility is that the cell cycle genes are controlled by other events which
occur during the cell cycle, and since senescent cells are not cycling, these
genes are not triggered by the proper series of events.
The full significance of cellular aging in vivo is not known. However, it
may be that small decrements in various systems act synergistically. For
example, a decline in lung capacity coupled with a decline in cardiac output,
a decline in hemoglobin content, or the oxygen-carrying capacity of the blood
could lead to a significant decline in the total oxygen available. It is clear
that loss of cell proliferative capacity in some organ systems can have serious
consequences for the organism as a whole. For example, if the cells lining
the vascular system were not able to proliferate in response to injury, den-
udation of the vascular system could result. This could cause thrombosis or
atherosclerosis.
On the other hand, it seems likely that the limited proliferative potential
of normal cells in vivo is a powerful inhibitor of tumorigenesis. Even when
some of the changes leading to tumor formation have occurred, the limited
proliferative potential of nonimmortalized cells severely limits the damage
caused by these potentially tumorigenic cells.
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Title: Sexual ethics
Author: Auguste Forel
Author of introduction, etc.: C. W. Saleeby
Translator: Ashley Dukes
Release date: October 18, 2023 [eBook #71898]
Language: English
Original publication: United Kingdom: The New Age Press, 1908
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*** START OF THE PROJECT GUTENBERG EBOOK SEXUAL

ETHICS ***
SEXUAL ETHICS
SEXUAL ETHICS
BY
AUGUST FOREL, M.D., PH.D., LL.D.

FORMERLY PROFESSOR OF PSYCHIATRY AT
AND DIRECTOR OF
THE INSANE ASYLUM IN ZURICH (SWITZERLAND)
WITH INTRODUCTION
BY
Dr. C. W. SALEEBY, F.R.S. Edin.
LONDON
THE NEW AGE PRESS
140 FLEET STREET
1908
Translated from the German by Ashley Dukes
INTRODUCTION
By Dr. C. W. SALEEBY, F.R.S. Edin.
T here is something absurd, as such, in a request for an

introduction by any one to the work of one of the greatest of
living thinkers, and something still more absurd in the fact that
Professor Forel should, at this date, need an introduction to any
intelligent audience in any civilised country, as it seems he does to
English readers; but if compliance with that request is at all likely to
increase, even by one, the number of his readers, it is a duty to
comply with it.
Not to consider his treatises on philosophy and psychology, nor his
long series of original and important researches on the senses and
lives of the social insects, Professor Forel has already given to the
world a volume entitled Die Sexuelle Frage—this has now been
published in English[A]—which is by far the best work on the sex
question in any language, and has actually received on the
Continent something like the recognition which is its due. The gist of
its teaching is to be found in this little treatise on Sexual Ethics, and
the reader who may find himself or herself unconvinced, or even
repelled, by the brief and dogmatic theses of the following pages,
may be earnestly counselled to read the larger work. Here, and in
that, Professor Forel deals—always from the loftiest moral
standpoint, the interests of human life at its highest—with the
question which must remain fundamental for man so long as he is
mortal, and with which the statesmen of the future will primarily
concern themselves, realising as they will, and as the “blind mouths”
called statesmen to-day cannot, that there is no wealth but life, that
the culture of the racial life is the vital industry of any people, and
must so remain so long as three times in every century the only
wealth of nations is reduced to dust and raised again from helpless
infancy. Professor Forel sees this question from the only standpoint
that is worthy of it. The sexual question is concerned with nothing
less than the life of this world to come. It is for this reason that every
productive sexual union should be a sacrament; it involves nothing
less than the creation of a human life—the most tremendous act of
which man or woman can be capable. It is the no less than sacred
cause of Eugenics or Race-Culture that gives the sexual life its
meaning and the dignity which it may rightly claim, and it is just
because the Swiss thinker sees this and never loses sight of it that
his work is so immeasurably raised above the ordinary discussions
of marriage, prostitution, venereal disease, and the like. His claim for
posterity on the ground of our debt to the past may be amplified by
the reflection that, in serving the racial life, and in making its welfare
the criterion of our sexual ethics, we are serving human beings as
real as we are ourselves, and tens or hundreds for units whom we
can serve to-day. There is always an interval—nine months at least
—and no one expects babies or politicians to associate cause and
effect over such abysses of time; but there are others who are
learning to think in generations, and Professor Forel will yet add to
their number.
[A] The Sexual Question. Rebman, Ltd.
In his criticisms of alcohol and the abuse of capital, Professor
Forel opposes himself to the most powerful of vested interests. Well,
if you invest your interests in any other bank than that of the laws of
life, you or your heirs will find that theirs is but a rotten concern. The
history of organic evolution is proof enough that the higher life and
the things which buttress it, “sagging but pertinacious,” will always
win through in the long run. As a direct enemy of human life, and
notably through its influence upon the sexual instinct, alcohol is
certainly doomed. If life is the only wealth, the manufacture of illth is
a process too cannibal to be permitted for ever.
Professor Forel speaks of subduing the sexual instinct. I would
rather speak of transmuting it. The direct method of attack is often
futile, always necessitous of effort, but it is possible for us to
transmute our sex-energy into higher forms in our individual lives,
thus justifying the evolutionary and psychological contention that it is
the source of the higher activities of man, of moral indignation and of
the “restless energy” which has changed the surface of the earth. As
directly interfering with this transmutation, the extent of which
probably constitutes the essential difference between civilised and
savage man, alcohol is the more to be condemned.
In what Professor Forel has to say regarding prostitution and the
ideal of marriage, he will win assent from all except the profligate
and those medical men who, in hideous alliance with the protozoon
of syphilis and the coccus of gonorrhœa, defend prostitution and
even acclaim it as the necessary complement to marriage. If there is
a stronger phrase than most damnable of lies to apply to such
teaching, here is certainly the time for its employment. On this
subject of prostitution, Professor Forel has said the last word in a
masterly chapter of Die Sexuelle Frage. In his praise of monogamy,
he is only echoing the stern verdict of the ages—delivered a
thousand æons before any existing religion was born or thought of,
and likely to outlast a whole wilderness of their dogmas. The
essence of marriage I would define as common parental care of
offspring, and its survival-value as consisting in the addition of the
father’s to the mother’s care. In the absence of parenthood, a sexual
association between man and woman is on the same plane as any
other human association; it means neither more nor less, and must
be judged as they are judged. It is when the life of the world to come
is involved that new questions arise—questions as momentous as is
the difference between the production of human life at its best and of
a child rotten with syphilis, or permanently blinded to the light as it
opens its eyes for the first time, or doomed to intelligence less than a
dog’s.
I, for one, have no shadow of doubt that the ideal of sexual ethics
will some day be realised, that pre-eminently preventable—because
contagious—diseases like syphilis and gonorrhœa will be made an
end of, that prostitution will disappear with its economic cause, that
we shall make parenthood the privilege of the worthy alone, and thus
create on earth a better heaven than ever theologians dreamed of in
the sky. “There are many events in the womb of Time which will be
delivered.” Individuals are mortal, and churches, and creeds, but Life
is not. Already the gap between moss or microbe and man is no
small one, and the time to come is very nearly “unending long.”
Uranium and radium will see to that.
C. W. SALEEBY.
SEXUAL ETHICS
T he two conceptions of morality and sexual life are frequently

confounded and expressed by the same term in the popular
usages of speech. The word “moral” is commonly used to mean
sexually pure, that is to say, continent; while the word “immoral”
suggests the idea of sexual incontinence and debauch. This is a
misuse of words, and rests upon a confusion of ideas, for sexuality
has in itself nothing to do with morality. It points, however, to the
undoubted fact that the sexual impulse, since it has other human
beings as its object, easily leads to moral conflicts within the breast
of the individual.
It will be convenient to discuss our subject under the two heads: I.
Of ethics in general; and II. Of sexual ethics in particular.
I. Ethics
Ethics is the science of morals. Morals may be said to consist of
two very distinct factors, which we will attempt to analyse:—
1. An instinctive sense, the conscience, sense of duty, or ethical
impulse, which says to us: “This shalt thou do, and that shalt thou
leave undone.” A person in whom it is highly developed experiences
satisfaction if he obeys the “voice of conscience,” and remorse if he
fails to do so.
2. The second factor of morals includes the objects of conscience,
that is, the things which conscience commands or forbids.
The great philosopher Kant founded upon the instinct of
conscience his Categorical Imperative, and held the further
investigation of its causes to be unnecessary. If the conscience says
“Thou shalt,” one must simply act accordingly. This is, in Kant’s
opinion, the absolute moral law, which bids or forbids an action
independently of any other consideration.
The further they progress, however, the more do reason and
science rebel against the conception of the Categorical Imperative.
Kant, great as he was, was not infallible. The imperative of the
conscience is in itself no more categorical and absolute than that of
the sexual impulse, of fear, of maternal love, or of other emotions
and instincts.
In the first place daily observation shows us the existence of
people born conscienceless, in whom the sense of duty is lacking,
who are aware of no “Thou shalt,” and in whose eyes other
individuals are merely welcome objects for plunder or inconvenient
hindrances. For these “ethically defective” persons there can be no
categorical imperative, because they have no conception of duty.
The ethical sense may exist in varying degrees of intensity. In
some persons the conscience is weak, in others strong; and there
are cases in which it is developed to an exaggerated and morbid
extent. People of this type suffer pangs of conscience over the
merest trifle, reproach themselves for “sins” which they have never
committed, or which are no sins at all, and make themselves and
others miserable. How can all this be reconciled with the absolute
moral law as stated by Kant?
The theory of the Categorical Imperative becomes even more
absurd when we consider the actions to which men are guided by
their consciences. The same habit—the drinking of wine, for instance
—may be for one man a matter of duty (for a Christian at the
Eucharist or for an officer at the toast of the King); for another (the
Mohammedan) it may be forbidden as a deadly sin. Murder, which is
certainly almost universally prohibited by conscience, is a “duty” in
time of war, and even for certain persons in the duel. Such instances
could be multiplied indefinitely.
We will presently state the profounder reasons which prove Kant’s
error; but we must first mention another source of pretended ethical
commandments. The religions exhibit a remarkable medley of
various products of human mystical phantasy and human emotions
which have crystallised and formed themselves into legends and
dogmas, and these latter have become interwoven with human
morals in such a fashion that they seem at first inextricable.
The instinct of fear and the lust for power, the hypertrophy of the
Ego and the ethical sentiments have here intermingled in a thousand
different ways. More especially we may mention the fear of the
unknown, of darker powers, and of death; the expansion of the
beloved Ego, which becomes idealised in the conception of
godhead, and then immortalised; the feelings of sympathy, antipathy
and duty towards other individuals, and so forth. The mysterious
powers which move the universe are then conceived as
anthropomorphic (personal) gods, or as one such God.
The next stage is the attribution of godlike qualities to man, which
flatters his vanity considerably, and gives him a sense of satisfaction.
As a result of this habit of thought, and assisted by the
hallucinations of highly imaginative, hysterical, or insane individuals,
there have developed the various conceptions of a direct intercourse
between the Godhead and man. Hypnotism and psychiatry, in the
respective cases of the sane and the insane, teach us how
extraordinarily sensitive the human brain is to such impressions.
In this way the legendary revelations, according to which God has
manifested himself directly and personally to certain individuals, and
dictated to them commandments for the guidance of Humanity, have
resulted.
In this, and in no other way, has come into existence the social
tyranny of religious dogmas. Certain men have made God in their
own image, and have, in the course of centuries, imposed their own
handiwork upon whole nations, mainly by means of the organising
ability of their more ambitious successors. Even to-day such
prophets frequently arise, both within and without the walls of lunatic
asylums. Each one declares that he alone possesses the true
revelation.
The divine injunctions vary considerably according to the different
religions, and are often mutually contradictory. Among them are
commandments relating to the Godhead which have nothing to do
with natural moral law, and yet are amalgamated with it. Some of
these are from the human point of view frankly immoral. Many, on
the other hand, represent the precepts of a more or less suitable
moral code, which varies according to the personal views of the
founder of the religion.
The Koran ordains polygamy and forbids the use of wine, while
modern Christianity allows the latter and ordains monogamy. Both
Moses and Mohammed, however, regard woman as subordinate to
man, and as his private property; a view which contradicts a higher
and at the same time a more natural moral law.
Mental science has now the hardihood to maintain, Kant and the
religious dogmas notwithstanding, that the moral law is completely
accessible to its investigations; that true human ethics can be
founded upon human nature alone; that the dogmas and
commandments of pretended revelation serve only to check a
progressively higher development of morals; and that the dogma
which holds out promises of heaven or threats of hell in the hereafter
is in its effect actually immoral, inasmuch as it seeks to regulate the
moral conduct of men by purely selfish motives—by the aid of a bill
of exchange upon the future life, so to speak.
In order to understand natural human ethics we must consider its

natural source, that is to say, the origin of the sense of duty or social
conscience.
The sense of duty is, as an inclination, inborn, and therefore
hereditary. It can indeed be developed or dulled by education, but it
cannot be acquired; and only diseases of the brain can destroy it
where it once clearly exists. What is actually inculcated or acquired,
as the case may be, is not the conscience, but the object towards
which it is directed, as is the case with the feeling of shame or
modesty. Just as the European woman is ashamed to exhibit her
bare legs, but not her face, while with the Turkish woman the reverse
holds true, so the objects of the conscience, according to acquired
local customs, can be absolutely opposed to one another, or at least
very different in their nature. They have, however, for the most part
certain features in common, which are suited to the requirements of
human nature. The reason for this we shall see below.
From what does conscience, or the sense of duty, arise? First of

all from a conflict between two groups of instinctive emotions allied
with instinctive impulses: (1) the group of so-called egoistic feelings
and impulses, directed towards self-preservation and self-
gratification; and (2) the group of sympathetic or altruistic impulses
directed towards the preservation and well-being of others.
If I feel sympathy or love for a person, an animal, or an object, I
suffer personally and feel displeasure as soon as the object of my
sympathy suffers or is endangered. Hence the words compassion
and sympathy (suffering with). I therefore seek to help the object of
my sympathy, to save him even at the risk of personal injury; and
thence the conflict arises. If my egotism triumphs I do not come to
his aid, or at most only do so if I risk nothing thereby. If, on the other
hand, my sense of sympathy is victorious, I sacrifice myself.
In the former instance I experience a feeling of dissatisfaction, the
feeling of neglected duty and of remorse; in the latter I have the
pleasurable sensation of duty fulfilled. And yet the nature of the
object matters little. Only the intensity of the sympathy, together with
the individual development of the conscience, determine the intensity
of the sense of duty in any given case. An insane person can feel the
most vehement sense of duty or remorse without any real object, or
as the result of entirely perverted conceptions.
As every living creature, particularly if it possesses a separate
nervous system, has the instinct of self-preservation, the conscience
therefore results directly from the conflict between this instinct and
the secondary emotions of altruistic sympathy. These latter are of
later origin, and have for the most part been evolved from the
attraction between the sexes (sexual love), or from the relationship
of parents to the offspring dependent upon them (parental love).
The first feelings of duty and of sympathy in the animal kingdom
are therefore confined to the family, and adapted to the preservation
of the species. They are also exclusive, and may only persist for a
short time (as in the case of cats), but frequently they are of lifelong
duration. The conjugal fidelity of certain apes and parrots is
exemplary.
But the necessity of protection against common foes brought
about in the case of many animals a ripening of the sense of
sympathy, and it became extended to whole groups, so that here
and there free communities (swallows, buffaloes, monkeys) have
resulted. Finally certain species have developed the senses of
sympathy and duty to such an extent that they have led to a
complete anarchistic Socialism, as is the case among wasps, bees,
and ants. Here the social sense has so far overcome both egotism
and altruism limited to a few individuals that it wholly dominates
them. The individual devotes his whole energy and labour to the
communal existence, and even sacrifices his life for this object. He
never, however, sacrifices his life for another single member of the
community, unless the latter is of primary importance for the
maintenance of the species. One worker-bee does not immolate
itself for another, but does so without hesitation for the queen and
the hive. It will even empty the whole contents of its stomach into the
queen bee’s mouth and starve in order to save her. The altruism of
the ants and the bees knows nothing of family affection or sexual
love; it is confined absolutely to the hive or nest. Different beehives
or ants’ nests are either inimical or indifferent to one another.
Nearer to man stand the higher mammals. Every one is aware of
the sentiments of sympathy and duty in the dog, for instance. In man
himself these affections are pre-eminently domestic, as may be seen
in the love of mother and child, husband and wife, father and son,
and in all the obligations thus contracted. But they also have a
considerable tendency to extend to other intimate objects or persons
with whom the individual frequently comes into contact—to friends,
animals, etc.
We can also observe this inclination among bees and ants, where
strangers are received into the hive or nest after a short period of
familiarisation. But among mankind the tendency always maintains a
strongly individual character. The result is on the one hand a
grouping into communities, such as castes, tribes, and nations; and
on the other a host of individual friendships and enmities.
This fundamentally individual character of the human sense of
sympathy rests primarily upon the fact that our nearest ancestors in
the animal world, the parents of the existing anthropoid apes, were
domestic and solitary, while our primeval ancestors lived in
numberless tiny communities, inimical to one another.
In this way there appeared among mankind instinctive and
exclusive impulses of sympathy and of duty, combined with intensely
selfish predatory desires. The extraordinary complexity of the human
brain is responsible for the strange many-sidedness of character
which resulted. For example, crime and heroism developed side by
side; child murder, parricide, rapine and robbery, slavery, war, and in
particular the vilest subjugation of woman as an article of commerce
or a beast of burden—these represent the fruits of egotism and its
attendant cunning and meanness. On the other hand we see self-
sacrifice, valour, heroic martyrdom, patriotism, sense of justice,
asceticism, pity for the weak, and persistent labour for the family and
the State, resulting as the fruits of the instinct of sympathy and the
social sense.
The primitive sense of duty, which arose from direct assistance
rendered to the object of sympathy, is now being enlarged by a
higher racial and individual development, and is, indeed, resolving
itself into a universal inclination to subdue egoistic instincts and
passions.
If from a sense of duty I do something which is wearisome or
dangerous, it is for the most part no longer out of direct sympathy
with the particular object. The primeval impulse (which led to conflict)
is becoming independent, and is taking the form of a higher and
secondary instinct, tending towards the suppression of baser desires
and weaknesses. And yet it is necessary, in order to prevent the
degeneration of this instinct, that the objects towards which it is
directed shall be ever more adequately and better suited to the
social welfare of the community.
From the above brief sketch, which is based upon the theory of
evolution and the researches of science, it is clear as the day that
moral laws can only be relative. They were always relative to the
family, to the tribe, to the fatherland; they must become relative to
mankind. The racial (that is, inherited and instinctive) social sense in
man is unfortunately very variable in individual cases. In the average
it is extremely weak and chiefly directed towards a few individuals.
Moreover, as the result of centuries of bad habits and ancient
prejudices, its objects are falsely or unsuitably taught in process of
educating children. Instead of the child’s sense of duty being
directed to the necessity of labour and social sacrifice for mankind as
a whole and posterity in particular, it is directed towards false codes
of honour, local patriotism, family exclusiveness, private property,
pretended divine commandments, and so forth.
The Earth is small, and human intercourse becomes more
extensive every year; the union of all civilised peoples into a single
great civilised community is inevitable. Ethics must, therefore, as far
as reason permits, be directed towards this object. We require

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Molecular and Oncogenetic

George E. Milo, M.S., Ph.D.

Bruce C. Casto, M.S., Sc.D.

Charles F. Shuler, D.M.D., Ph.D.

CRC Press is an imprint of the

Reissued 2018 by CRC Press

© 1992 by Taylor & Francis

No claim to original U.S. Government works

Trademark Notice: Product or corporate names may be trademarks or registered trademarks,

ISBN 13: 978-1-138-10503-4 (hbk)

George E. Milo, B.A., M.S., Ph.D., is Professor of Medical Biochem-

Bruce C. Casto, M.S., Sc.D., is Director of Research for Environmental

Charles F. Shuler, D.M.D., Ph.D., is Assistant Professor in the Center

William M. Baird, Ph.D. Frank C. Cuttitta, Ph.D.

Bruce C. Casto, M.S., Sc.D.

IN VITRO CELLULAR AGING AND IMMORTALIZATION

II. In Vitro Cellular Aging is Dominant in Somatic Cell Hybrids 3

III. Cellular Aging Is an Active Process 6

Carcinogenesis has become widely accepted as a multistep process2 (see

II. IN VITRO CELLULAR AGING IS DOMINANT IN

A. LIMITED IN VITRO LIFE-SPAN OF NORMAL CELLS

B. HYBRIDS BETWEEN NORMAL AND IMMORTAL CELLS

immortalization was due to dominant changes in the cellular genome. Many

C. FUSION OF IMMORTAL CELLS WITH OTHER IMMORTAL

D. MICROCELL HYBRID EXPERIMENTS

III. CELLULAR AGING IS AN ACTIVE PROCESS

B. MEMBRANE-ASSOCIATED DNA SYNTHESIS INHIBITORS OF

inhibitor by treating the cells with cyclohexamine or puromycin at concen-

C. INHIBITION OF DNA SYNTHESIS BY POLY(A)+

The experimental results reviewed here indicate that cellular aging is an

DNA synthesis by senescent cells. The mechanism by which this inhibitor

1 1 . Swim, H. E. and Parker, R. F., Culture characteristics of human fibroblasts propagated

Title: Sexual ethics

Author: Auguste Forel

Author of introduction, etc.: C. W. Saleeby

Translator: Ashley Dukes

Release date: October 18, 2023 [eBook #71898]

Original publication: United Kingdom: The New Age Press, 1908

Credits: Produced by Tim Lindell, Donald Cummings, and the Online

*** START OF THE PROJECT GUTENBERG EBOOK SEXUAL

AUGUST FOREL, M.D., PH.D., LL.D.

Dr. C. W. SALEEBY, F.R.S. Edin.

T here is something absurd, as such, in a request for an

T he two conceptions of morality and sexual life are frequently

In order to understand natural human ethics we must consider its

From what does conscience, or the sense of duty, arise? First of

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