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books you buy, genic and parasitic organisms, understanding their complex life
cycles and their modes of transmission is critical to understand-
the greater your
discount!
ing their effects on individuals and how disease outbreaks occur
in ecological systems. Communication between the pathogen
and the host organism occurs in the course of infection and
Genetic and
THE CONTENT
Energy Physics
involves the disruption of normal cell function. Finally, epide-
miology is briefly discussed, using the case of severe acute Pathogenic
Diseases
respiratory syndrome (SARS). Data are used to describe how
Engineering
the disease may have originated and evolved to infect humans,
Biotechnology
and how it spread relatively quickly and almost caused a global
onCells
Biology pandemic. Understanding how disease outbreaks occur in eco-
Mathematics logical systems is critical to controlling the spread of disease.
Chemistry
Christopher J. Paradiseis professor of biology and environ-
mental studies at Davidson College. He teaches introductory
THE TERMS biology, ecology, entomology, and topical seminars on ecotoxi-
Perpetual access cology and renewable natural resources. He also occasionally
for a one time fee leads a study abroad program in India. His research evaluates
anthropogenic factors that influence insect biodiversity at a
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A. Malcolm Campbell
Effects of Genetic and
Pathogenic Diseases
onCells
Effects of Genetic and
Pathogenic Diseases
onCells
10 9 8 7 6 5 4 3 2 1
Keywords
mutations, disease, point mutation, motor neuron disease, familial disease,
normality, emerging disease, cytokines, rice blast fungus, pandemics, sickle-
cell disease, epidemiology, perfection, pathogen, vector-borne disease, Lyme
disease, macrophage, neutrophil, parasite, Giardia, outbreak, SARS, disease
reservoir
Contents
Preface...................................................................................................ix
Acknowledgments....................................................................................xi
Introduction.........................................................................................xiii
Chapter 1 Genetic Diseases Affect Cells and Organisms.....................1
Sickle-Cell Disease is Caused by a Single Point
Mutation........................................................................1
Multiple Mutations may Cause a Single Disease.................9
Ethical, Legal, Social Implications: Normality
andPerfection...............................................................12
Chapter 2 Pathogens Affect Cells and Organisms.............................15
Lyme Disease Is Caused by a Vector-Borne Bacterium......15
Ethical, Legal, Social Implications: There are Several
Issues with Using Animals in Research..........................22
Fungi can be Pathogens of Plants.....................................24
Chapter 3 Parasites can Survive in More than One Host Species.......29
Chapter 4 Diseases Spread Through Populations of Susceptible
Hosts...............................................................................41
Conclusion............................................................................................51
Glossary................................................................................................53
Index....................................................................................................55
Preface
This book about the effects of genetic and pathogenic diseases on cells is
part of a thirty book series that collectively surveys all of the major themes
in biology. Rather than just present information as a collection of facts,
the reader is treated more like a scientist, which means the data behind the
major themes are presented. Reading any of the thirty books by Paradise
and Campbell provides readers with biological context and comprehen
sive perspective so that readers can learn important information from a
single book with the potential to see how the major themes span all size
scales: molecular, cellular, organismal, population and ecologic systems.
The major themes of biology encapsulate the entire discipline: informa
tion, evolution, cells, homeostasis and emergent properties.
In the twentieth century, biology was taught with a heavy emphasis
on long lists of terms and many specific details. All of these details were
presented in a way that obscured a more comprehensive understanding.
In this book, readers will learn about the effects of genetic and patho
genic diseases on cells and some of the supporting evidence behind our
understanding. The historic and more recent experiments and data will
be explored. Instead of believing or simply accepting information, read
ers of this book will learn about the science behind the effects of genetic
and pathogenic diseases on cells the way professional scientists dowith
experimentation and data analysis. In short, data are put back into the
teaching of biological sciences.
Readers of this book who wish to see the textbook version of this
content can go to www.bio.davidson.edu/icb where they will find
pedagogically-designed and interactive Integrating Concepts in Biology
for introductory biology college courses or a high school AP Biology
course.
Acknowledgments
Publishing this book would not have been possible without the generous
gift of Dr. David Botstein who shared some of his Breakthrough Prize
with co-author AMC. Davids gift allowed us to hire talented artists (Tom
Webster and his staff at Lineworks, Inc.) and copyeditor Laura Loveall.
Thanks go to Kristen Mandava of Mandava Editorial Services for project
management and guidance. In particular, we are indebted to Katie Noble
and Melissa Hayban for their many hours and attention to detail.
Kristen Eshleman, Paul Brantley, Bill Hatfield and Olivia Booker
helped us with technology at Davidson College. We are grateful to ad
ministrators Tom Ross, Clark Ross, Carol Quillen, Wendy Raymond,
Verna Case, and Barbara Lom who had confidence in us and encouraged
us to persist despite setbacks along the way.
Thanks to my wife Amy Brooks for her constant support during the
development of this textbook, and my daughter Evelyn for her endless
energy. Thanks to Malcolm Campbell for his steadfast resolve and opti
mism. Without him, this book would not exist. Thanks to collaborator
Laurie Heyer for taking my sometimes half-baked math ideas and turn
ing them into powerful and elegant Bio-Math Explorations. I learned
a lot from both of them. While the math is largely absent from this
book, our collaboration with her made this a better book. Nancy Stamp
at Binghamton University, and Bill Dunson and Richard Cyr at The
Pennsylvania State University influenced me greatly in how I think as
a scientist and approach my teaching. Finally, I thank my students in
Integrated Concepts in Biology II, who enthusiastically participated
in our experiment to redesign introductory biology, starting with the
text and ending with a new approach to teaching biology.
Introduction
In the latter half of the seventeenth century Anton van Leeuwenhoek
used a simple microscope to become one of the first people to view liv
ing cells in the form of blood, rainwater, and algae. He viewed microbes
and cells of multicellular creatures. Much later, in the mid-nineteenth
century, other scientists observed plant and animal tissues under micro
scopes. These scientists discovered that the parts are made of cells and
that cells could be considered individual units within multicellular organ
isms. This led Henri Dutrochet to state that the cell is the fundamental
element of organization and Rudolph Virchow, a German physician, in
1855 to state that all living cells come only from other living cells. The
observations of these scientists led to the development of Cell Theory,
which states that all organisms are made from one or more cells, cells
only arise from preexisting cells, and the cell is the smallest form of life.
Themes of the cell concept also include: maintenance of an internal en
vironment, structure defines function, and communication. Here how
diseases can have a variety of effects in organisms will be examined. A
single population of a unicellular pathogenic organism can interact with
cells or tissues in one or more host species, or genetic diseases can affect
cells and tissues in multicellular organisms.
CHAPTER 1
sixty to forty. Pauling and his colleagues concluded that individuals with
mild sickling have both types of hemoglobin and are heterozygous for the
gene that produces hemoglobin. Sickle-cell anemia and normal individu
als are both homozygous.
In the late 1950s Vernon Ingram began the process of trying to deter
mine the exact nature of the differences between normal and sickle-cell
disease hemoglobins. He predicted that the sickle-cell hemoglobin had
one less carboxylic acid group in its protein structure, which would ac
count for the difference in charge observed by Paulings group.
Ingram denatured purified hemoglobin. He then digested it with
trypsin, an enzyme that breaks down proteins, predominantly wherever
the amino acids lysine and arginine are found, except when either is fol
lowed by proline. The digestion produced shorter peptide chains that
were reproducible, because each type of hemoglobin would always be
broken into chains of particular amino acid composition when exposed
to trypsin. If there were differences between the sets of chains, then those
proteins could be identified as having different amino acid sequences.
Ingram performed both electrophoresis and chromatography on
these samples to separate the peptide chains in two dimensions. Peptides
are separated by both their charges in an electric current and their mobil
ity in a solvent gradient. Small peptides move faster with the leading edge
of the solvent. The proteins are then dyed, and the resulting image is the
peptide fingerprint of a protein. Ingram carefully traced the blotches of
peptides from the filter paper and numbered equivalent pairs.
The two hemoglobin fingerprints could be almost completely super
imposed on each other. One peptide (labelled #4) was in different posi
tions in normal and sickle-cell hemoglobin. It appears along the set of
peptides that are slightly positively charged. The altered peptide has a
stronger positive charge in sickle-cell hemoglobin relative to normal he
moglobin, which caused it to migrate toward the negatively charged elec
trode during electrophoresis. Superimposition of other peptides indicates
that the amino acid sequences were the same.
Ingram isolated peptide #4 from the two hemoglobins. He broke it
down into smaller chains and performed his two dimensional fingerprint
ing technique on those peptides. He knew the amino acids that were
present in these peptides, based on other analyses, and that there was
Genetic Diseases Affect Cells and Organisms 5
more glutamic acid in the normal hemoglobin chain and more valine in
the sickle-cell hemoglobin chain. In determining the amino acid chain for
this peptide, recall that trypsin cuts proteins wherever it finds a lysine or
an arginine. Because arginine was not found in this peptide, lysine must
be at the end of each chain. Using empirical data and knowledge of the
structures and charges of each amino acid, Ingram deduced the peptide
structure of each spot on the fingerprints.
The two longest non-overlapping sequences for either peptide chain
are four and five amino acids in length, making the minimum chain
length nine. Taking into account the overlap in sequences that he found,
Ingram lined up the sequences as in Table 2.
Ingram showed that in this particular peptide chain, the seventh posi
tion was altered, such that in sickle-cell hemoglobin a valine was substi
tuted for a glutamic acid. Glutamic acid has a carboxylic acid on its side
chain, which would make it more negatively charged than the valine, or
in the case of the full nine amino acid sequence, the negative charge on
the glutamic acid balances positive charges in the other amino acids to
make it more neutral than the peptide from sickle-cell hemoglobin.
Ingram correctly deduced that the sickle-cell peptide had one less car
boxylic acid group than normal hemoglobin. Scientists later determined
that Ingram misinterpreted the fingerprintthe H-V-L-L sequences
turned out to be V-H-L. Scientists now know that the amino acid substi
tution occurs at the sixth position, not the seventh; the corrected normal
sequence is V-H-L-T-P-E-E-K, and the corrected sickle-cell sequence is
V-H-L-T-P-V-E-K. A point mutation in the DNA sequence of individu
als with sickle-cell anemia leads to the altered hemoglobin. Sickle-cell ane
mia became the first genetic disorder whose molecular basis was known.
Recall that Pauling and his colleagues were aware that sickling occurred
primarily when oxygen content is low. Perutz and Mitchison (1950) ex
amined red blood cells from sickle-cell disease and normal patients. When
hemoglobin was deoxygenated, they found that sickled cells exhibited
physical properties similar to crystals, but normal cells did not. The re
searchers concluded that sickle-cell hemoglobin crystallizes in the absence
of oxygen. The scientists further went on to test the solubility of oxygen
ated and deoxygenated hemoglobins in solutions of varying ionic concen
trations (Table 3). The scientists determined the maximum concentration
of each hemoglobin in each condition that could be dissolved in solution.
The point mutation that produces sickle-cell hemoglobin creates a
protein with different physical properties from normal hemoglobin. When
saturated with oxygen, it behaves like normal hemoglobin, having the
same solubility. But when it has lost some or all of its oxygen, as it does
in venous blood, it begins to precipitate and crystallize and its solubility
declines. At any ion concentration, oxygenated hemoglobin has higher
solubility than deoxygenated hemoglobin. At ionic strength of 5.0, the
solubility of either type of oxygenated hemoglobin is 6.3 grams per liter,
dull and boring. In some countries, like the United States, people value
individuality of thought and expression, and this could be lost in a world
where everyone thought and looked alike.
These examples illustrate the extreme. With more than 7 billion peo
ple on the planet, it is safe to say that we will remain diverse in looks and
thought for many years to come. As long as that diversity of thought
remains, then what it means to be perfect will vary, and humanity may
never converge on a single way of thinking and looking.
Bibliography
Allison AC: Protection afforded by the sickle-cell trait against subtertian
malaria infection, Br Med J 1(4857):290294, 1954.
Ingram VM: Gene mutations in human haemoglobin: the chemi
cal difference between normal and sickle-cell haemoglobin, Nature
180(4581):326328, 1957.
Ingram VM: Abnormal human haemoglobins. I. The comparison of nor
mal human and sickle-cell haemoglobins by fingerprinting, Biochim
Biophys Acta 28(3):539545, 1958.
Ingram VM: Sickle-cell anemia hemoglobin: the molecular biology of
the first molecular diseasethe crucial importance of serendipity,
Genetics 167(1):17, 2004.
Kwiatkowski TJ Jr, Bosco DA, Leclerc AL, et al.: Mutations in the FUS/
TLS gene on chromosome 16 cause familial amyotrophic lateral scle
rosis, Science 323(5918):12051208, 2009.
Pauling L, Itano HA, Singer SJ, et al.: Sickle-cell anemia, a molecular dis
ease, Science 110(2865):543548, 1949.
Perutz MF, Mitchison JM: State of haemoglobin in sickle-cell anemia,
Nature 166(4225):677679, 1950.
CHAPTER 2
Much can be learned about how cells function by learning how vari
ous environmental factors disrupt cell function and homeostasis. In this
chapter, how some pathogens cause disease by disrupting cell functions
will be examined. Pathogens can have similar effects to those of genetic
diseases, in that they can disrupt cell function, which disrupts function
at the organismal level, depending upon what specific cells pathogens are
attacking. One pathogen that affects humans and another one that affects
plants will be used to illustrate general effects of pathogens.
Although the exact role of OSP-C in the infection cycle is still unknown,
it may be required for this pathogenic bacterial cell to evade the immune
system during the early phase of infection and spread through the blood
stream of the host.
The immune system of mammals consists of millions of cells of several
different types that play various roles in protecting the individual from
pathogens and other foreign substances. These cells move about the body,
protecting the individual. Their development, movement, and adhesion
to foreign invaders are determined by chemical signaling. Molecules used
in chemical signaling in the immune and other systems are cytokines,
small proteins, peptides, or glycoproteins released by cells involved in cell-
cell interaction and communication.
Chemokines are small protein cytokines that have receptors on spe
cific cells. When the chemokine binds to the receptor, it elicits a response
from the cell. For instance, leucocytes, or white blood cells, may respond
to a gradient of chemokine molecules by moving toward areas of higher
concentration. The release of the chemokine from tissues during bacte
rial infection or inflammation attracts leucocytes to the area of infection,
where they perform their function of attacking the invaders. Neutrophils
and monocytes are phagocytic leucocytes involved in the inflammatory
response. Phagocytes are white blood cells that ingest microbes, other
cells, and foreign particles.
In an experiment on early infection of mammalian hosts, Xu and his
colleagues investigated the ability of B. burgdorferi to evade early detec
tion, and thus establish a chronic infection. Recall the bullseye target
shaped swollen area commonly associated with the early phase of Lyme
disease and B. burgdorferi infection. This is an inflammatory response,
meaning the immune system is responding to the infection; lymphocytes
infiltrate infected tissues after being attracted to the tissues by chemo
kine signals. Several types of white blood cells are attracted to the site of
infection, yet there is a noted lack of neutrophils in B. burgdorferi infec
tions. This is unusual because neutrophils are the most common type of
lymphocyte and are often the first to arrive at sites of bacterial infection.
Clearly there is an inflammatory process in B. burgdorferi infection, yet
the immune response does not effectively clear the pathogen from the
body of the host as it should during inflammation. Xu and colleagues
Pathogens Affect Cells andOrganisms 19
= neutrophils
= monocytes
80
60
40
20
0
Bb ( control) Ec w/KC (+ control) Bb w/KC
Figure 1 KC activity of cell culture supernatants. The first
200leucocytes observed in each sample were identified as either
neutrophils or monocytes. The percentage shown is the overall
percentage for all mice sampled. Bb - B. burgdorferi; Ec - E. coli;
KC- chemokine.
Source: From Xu et al., 2007, Table 3.
were all found within the first 6 hours, but by 16 hours the latter two cell
types had disappeared and eosinophils appeared and stayed for 72 hours.
In the genetically engineered B. burgdorferi injection sites, the researchers
found large numbers of neutrophils within the first 6 hours whose num
bers peaked at 16 hours and were still present at 24 hours. By 72 hours
the neutrophils had disappeared.
Other mice that received only one high dosage injection of either KC-
producing B. burgdorferi or normal B. burgdorferi were killed and exam
ined 30 days post-injection for B. burgdorferi in the heart, joints, and skin
(Table 5A). Finally, one last set of mice was given one injection each, but
the dose of B. burgdorferi cells varied. Xu and his colleagues performed
this test to determine the lowest dose that infected mice. After 30 days,
mice were killed and inspected for infection in the heart, joints, and skin,
as with the other set of mice (see Table 5B).
Pathogens Affect Cells andOrganisms 21
engineered B. burgdorferi below 103 cells, because no mice below 106 cells
showed any evidence of successful invasion. By testing the mice in order
from high to low dose, they were able to reduce the total number of mice
tested because it would be unlikely that any mouse given a dose lower
than 103 would react.
It is unknown how B. burgdorferi normally evades the hosts immune
system, but the mechanism results in lower attraction of neutrophils to
the site of infection. Tick saliva has been shown to reduce neutrophil
function and attraction to a tick bite. Perhaps B. burgdorferi takes advan
tage of this mechanism that ticks use to evade the immune system. There
is a normal inflammatory response that causes neutrophils to be recruited
early on in the infection, but neutrophils disappeared quickly in mice
infected with normal B. burgdorferi, allowing the bacteria to establish
within the host. The high numbers of macrophages, which kill bacteria,
suggest that the bacterial infection would be eliminated, but for unknown
reasons, the macrophages were not effective against B. burgdorferi.
After the initial infective population of B. burgdorferi is established,
the bacteria migrate to joints, the heart, the nervous system, and other
tissues. Once they establish populations in other tissues, Lyme disease
manifests itself in many ways. Researchers have found that these bacteria
evade further immune responses by decreasing expression of OSPs. These
OSPs may be targeted by antibodies, and antibodies cannot recognize
invaders without the protein present. The bacteria can hide in the extra
cellular matrix and tissues where the immune system may not be able to
respond effectively. Disruption of the immune system by the pathogen
causes chronic inflammation even after the pathogen is gone from the
system, and may be an example of pathogen-induced autoimmunity.
ranging in the millions around the world, is much more than is needed
and is unacceptable from the animal rights advocates perspective. The
more animals held in captivity, the greater the chance that abuse, mis
treatment, or cruelty will occur. Advances in growing cell and tissue cul
tures will allow experiments to be done without the use of animals. Even
if we argue that mice are fairly closely related to humans (and they are
in the sense that both are mammals), the results from studies such as the
Lyme disease experiments may not be a useful guide to what is happening
when B. burgdorferi infects a human. If humans used only animals that
were very closely related to them (such as, chimpanzees), they are being
even more unethical, because chimpanzees are intelligent and sentient be
ings. If the humans dosed with malaria were not experimented on unless
they gave their consent and humans and other animals have equal value,
then humans cannot justify experimenting on an animal that is not able
to consent to the procedure.
Once the external concentration got too high, the cells collapsed and
lost their internal pressure. At that point cells were no longer able to
penetrate the surface on which they adhered, and that told the researchers
that external was close to internal pressure. Because not all cells behave
the same when exposed to a particular environmental condition, the sci
entists used the point at which 50% of the cells collapsed.
The scientists found that internal pressure increased prior to polymer
penetration. The longer the cells incubated prior to exposure to a PEG
solution, the greater the external pressure needed to be in order to cause
a high degree of cell collapse. In other words, over time the internal pres
sure builds up in these infection cells and thus in order to cause collapse
a higher external pressure is needed.
The scientists next let infection cells attach to and attempt to penetrate
the six different polymer sheets for different lengths of time, and then 100
cells were tested for their ability to penetrate the substrate. High pressure
in infection cells allowed penetration of even the hardest substrate, but
the harder the substrate, the more incubation time was required.
Howard and his also colleagues determined the pressure that inhib
ited penetration of substrates; if penetration occurs at a particular external
pressure, this indicates that internal pressure is high enough to penetrate
the particular substrate. To test this, infection cells were grown in water
for 18 hours, after which the water was replaced with various concentra
tions of a very large PEG. When the external osmotic pressure increased,
penetration was inhibited severely even for hard surfaces.
From these clever experiments, Howard and his colleagues determined
that pressure built up in infection cells and that the longer pressure built
up, the harder the surface was that could be penetrated. The enormous
pressure was shown to allow penetration of plant cuticles. Once inside the
plant cell, the invading fungus swells and fills the cell. As the outside struc
ture grows, neighboring epidermal cells of the rice plant are penetrated by
more cells. A few days after multiple penetration events, visible symptoms
of infection can be observed. As the infection grows, the fungus disrupts
cells in the epidermis of the rice leaf, and as multiple areas grow together a
gray lesion develops where fungal spores are produced.
Individual hosts can be infected by pathogens and that pathogens have
unique structures that function to disrupt host cells and allow invasion of
28 EFFECTS OF GENETIC AND PATHOGENIC DISEASES ON CELLS
the host. Host cells may no longer be able to maintain homeostasis and
normal function when invaded by pathogens, which can cause disease or
death. Pathogenic cells may communicate with host cells, causing them
to malfunction, or the pathogens can invade host cells. Either way, cell
function is often disrupted, leading to problems for the entire organism.
In the next chapter, more about parasites will be explored, in particular,
how it is that parasites can survive in multiple host species.
Bibliography
Grimm D, Tilly K, Byram R, et al.: Outer-surface protein C of the Lyme
disease spirochete: a protein induced in ticks for infection of mam
mals, Proc Nat Acad Sci 101(9):31423147, 2004.
Howard R, Ferrari MA, Roach DH, et al.: Penetration of hard substrates
by a fungus employing enormous turgor pressures, Proc Nat Acad Sci
88(24):1128111284, 1991.
Xu Q, Seemanapalli SV, Reif KE, et al.: Increasing the recruitment of
neutrophils to the site of infection dramatically attenuates Borrelia
burgdorferi infectivity, J Immunology 178(8):51095115, 2007.
CHAPTER 3
flagella
had experienced diarrhea in the past 2 months and, for a subset of each
of the three groups of children, obtained fecal samples on three separate
occasions to test for Giardia (Figure 3A).
Part of each fecal sample was preserved in 5% formalin and exam
ined microscopically for Giardia cysts, capsules, or resistant covers formed
around microorganisms in resting stages. Another part of each sample was
preserved, stained, and examined microscopically. The staining method they
used is rapid; it preserves the microscopic organisms and allows researchers
to distinguish among different amoebae and flagellates. The scientists also
interviewed the parents and collected fecal samples from any 1- to 2-year-
old siblings of children enrolled in the day care center (see Figure 3B).
Giardia was much more prevalent in the children who attended the
day care center with the diarrheal outbreak, even though there were signifi
cant percentages of children at the other day care centers with both diar
rhea and Giardia. This suggests that Giardia is at least a partial cause of the
outbreak at the day care center. The lower percentages of Giardia-free chil
dren with various clinical symptoms suggest a strong relationship between
this pathogen and the diarrheal outbreak. However, other pathogens that
Parasites can Survive inMore than One Host Species 31
70
50
40
30
20
10
0
A day-care day-care surrounding
with outbreak without outbreak community
20
= Giardia (+) family members
= Giardia () family members
percentage in children
15
10
0
B Giardia (+) child Giardia () child
Figure 3 Prevalence of diarrhea and Giardia in a human population.
A, Percentages in children from day care centers and surrounding
community. B, Giardia positive and negative family members with
children in day care center with diarrheal outbreak.
Source: From Black et al., 1977, Table 1 and text.
60 p < 0.04
50
= percent Giardia (+) children
percentage of children
20 NS
10 NS NS NS
NS
NS
0
ea
ys
ng
ps
ea
ou th
ve
os
nc
uc i
da
am
m w
s
rh
iti
us
fe
tl
le
m
ar
d/ ea
na
gh
10
tu
cr
vo
di
oo rrh
fla
ei
r>
bl ia
fo
d
ea
rh
ar
di
30 Giardia prevalence
25
= first survey
percentage of children
= second survey
20
15
10
0
percentage with cysts percentage with trophozoites
Figure 5 Percentage of children in day care centers with Giardia
cysts and trophozoites.
Source: From Pickering et al., 1984, Table 1.
34 EFFECTS OF GENETIC AND PATHOGENIC DISEASES ON CELLS
and weight loss (55.9). Many symptoms observed in children in day care
centers experiencing Giardia outbreaks appeared in the campers. The
onset of new cases (especially the spike at the end of the camping trip)
strongly suggests that the outbreak was caused by a parasite that infected
the campers while on the trip.
The researchers retraced the steps of the campers and searched for
Giardia cysts in stream water from which the campers drank. All campers
drank from the same water source, but they broke up into four groups to
search for food, with each group consuming what they had collected. The
scientists filtered stream water to concentrate any cysts on the filters. They
found no Giardia cysts. The only common source that all campers were
exposed to was the water, because groups of campers ate food that was not
shared among them, and campers from all four food-collecting groups
got sick. However, the researchers did not find cysts in the stream or in
beavers or muskrats living in streams. Thus there is no direct evidence
of Giardia in the water or in wild animals using the stream as habitat.
However, cysts are difficult to find in contaminated water, because they
are strongly diluted in such sources. Cysts can live outside of a host for up
to a year as shown previously, and the stream water was the only source
of water.
The scientists also examined the small and large intestines of ten
beavers and seven muskrats trapped in the drainage area, and found no
Giardia in the animals. Although evidence of Giardia in wild animals living
in the area might have been expected, the sampled beavers and muskrats
were a small proportion of these populations, and these individuals might
happen to not be infected or the species are not suitable hosts for Giardia.
Brett Dunlap and Monte Thies trapped and killed beaver (Castor ca-
nadensis) and nutria (Myocastor coypus), two semi-aquatic rodents, to pre
vent them from causing damage to human property. The beaver is native
to North America, whereas the nutria is an exotic species indigenous to
South America. The scientists collected a small intestinal and a fecal sam
ple from the large intestine from 100 beavers and 30 nutria. The samples
were preserved and later examined for the presence of Giardia using stain
ing and microscopy techniques similar to those used earlier (Figure 6).
Giardia is common in many mammal species, so it is not surprising
to find it in beavers and nutria. Dunlap and Thies did not distinguish
38 EFFECTS OF GENETIC AND PATHOGENIC DISEASES ON CELLS
100
80
sampled with Giardia
percent of animals
60
40
20
0
beaver nutria
Figure 6 Prevalence of Giardia in beaver and nutria captured
ineastTexas.
Source: Data from Dunlap and Thies, 2002, Tables 1 and 2.
Bibliography
Barbour AG, Nichols CR, Fukushima T: An outbreak of giardiasis in a
group of campers, Am J Trop Med Hyg 25(3):384389, 1976.
Parasites can Survive inMore than One Host Species 39
Black RE, Dykes AC, Sinclair SP, et al.: Giardiasis in day-care centers:
evidence of person-to-person transmission, Pediatrics 60(4):486491,
1977.
Campbell JD, Faubert GM: Comparative studies on Giardia lamblia
encystation in vitro and in vivo, J Parasitol 80(1):3644, 1994.
Craft JC: Experimental infection with Giardia lamblia in rats, J Infect Dis
145(4):495498, 1982.
Dunlap BG, Thies ML: Giardia in beaver (Castor canadensis) and nutria
(Myocastor coypus) from east Texas, J Parasitol 88(6):12541258, 2002.
Pickering LK, Woodward WE, DuPont HL, et al.: Occurrence of Giardia
lamblia in children in day care centers, J Pediatr 104(4):522526, 1984.
CHAPTER 4
envelope
ssRNA + nucleocapsid
matrix
spike
brought from rural outlying cities to Guangzhou for better medical care.
Many of these initial patients were known to be related to one another.
One SARS patient, admitted on December 17, 2002, was a chef who
worked at a restaurant in Shenzhen. This chef came into regular contact
with live caged animals used as exotic game food. He was the second
confirmed case of SARS.
The scientists also studied 55 patients admitted to a hospital in
Guangzhou, Guangdong Province, in early 2003 with atypical pneumonia.
These patients ran fevers for an average of 11.4 days ( 6.8 days, standard
deviation) and developed pneumonia within 4 days of being admitted.
Forty-one of 55 were known to have had definitive contact with another
SARS patient, and 27 were healthcare workers. Guan and his colleagues
examined mucus and blood serum samples for the SARS coronavirus
antibodies. For 22 of the patients, the scientists tested paired samples,
the first either 3 to 5 or 7 to 10 days after disease onset, and the second
15days after onset. Twenty (91%) of these patients tested positive for
SARS antibodies, and 28 (85%) of the remaining 33 patients tested only
once also were found positive for SARS antibodies. Four of the pneumo
nia patients who did not test positive for the SARS coronavirus antibod
ies were later discovered to have influenza. The researchers also tested
60healthy adults; none tested positive for SARS coronavirus antibodies.
The study of the initial outbreak is an important part of epidemiology
and can pinpoint the origin of an epidemic, identify modes of transmis
sion, and identify possible origins of the pathogen. In the southern China
SARS outbreak of 2002, several observations were made by epidemiolo
gists that were clues to origins and modes of transmission. One observa
tion was that the second known SARS patient was a chef who came into
regular contact with live caged animals. The epidemiological investigation
discovered that his wife, two sisters, and seven healthcare workers all be
came infected from him.
In fact, 34% of all patients in the outbreaks in the initial six cities were
healthcare workers. A patient in Guangzhou who spent only 18 hours in a
hospital in Zhongshan infected 30 healthcare workers in Zhongshan and
Guangzhou. This patient also infected 19 family members or relatives in
Guangzhou. This large number of infections in relatives and healthcare
workers gave rise to the Guangzhou outbreak. One physician infected
44 EFFECTS OF GENETIC AND PATHOGENIC DISEASES ON CELLS
during this time traveled to Hong Kong and became the source of the
SARS outbreak that occurred later in Hong Kong. Because the outbreaks
were characterized by infections in hospitals and family units, Guan and
his colleagues concluded that transmission occurs through close contact
with infected patients. The spread from city-to-city occurred because of
transport of patients to better hospital facilities and movement of workers
from city-to-city. This is comparable to a parasite like Giardia, which is
transmitted through fecal-oral contact or contaminated water.
Close person-to-person contact appears to be the mode of transmis
sion for SARS, where coughing or sneezing spreads small droplets up to
about a meter from infected persons, and disperses the virus. The virus
only needs to land on the mouth, nose, or eyes of a nearby person for
them to become infected. If droplets land on another persons hands, or
if an uninfected person touches a surface contaminated with droplets
and then touches their mouth, nose, or eyes, they can become infected.
Close contact may also mean kissing or hugging between an infected
and uninfected person. These are thought to be the most likely routes of
transmission.
Most patients with atypical pneumonia that were tested for the SARS
coronavirus had it as shown earlier. Several were infected with a strain of
the influenza virus. Given the large percentage of patients in this study
with the SARS coronavirus, Guan and his colleagues concluded that the
outbreaks were caused by this newly emerging viral pathogen. The scien
tists were able to conclude that the pathogen was emerging because none
of the healthy patients had antibodies to the SARS coronavirus, which
suggests that the virus was not previously present in this human popula
tion. The researchers, in attempting to track down the origin of the SARS
coronavirus, focused on the second known SARS patient, the chef, and
the observation that he came into contact with exotic game animals. In
China and many other countries, wild animals are caught or raised in cap
tivity to be sold live at markets for human consumption. Some wild game
animals are considered a delicacy in southern China and are bought by
chefs for preparation at their restaurants. Other diseases are also known
to spread from animals to humans.
Guan and his colleagues set out to test the hypothesis that the SARS
coronavirus was present in wild animals sold at southern Chinese markets.
Diseases Spread Through Populations of Susceptible Hosts 45
The researchers tested a variety of both domestic and wild mammals that
might be carrying the SARS coronavirus in Guangdong Province. The 25
animals sampled were obtained from a live animal retail market in Shen
zhen and included seven wild and one domestic mammal species.
The animals came from different areas of Guangdong and had been
kept in separate storehouses prior to coming to market. Only a few ani
mals of each species were kept by any one stall owner. Guan and his
colleagues obtained nasal, fecal, and blood (when possible) samples from
animals housed in different stalls. The sampled animals were confirmed
to have no overt symptoms of disease by a veterinarian. The researchers
used reverse transcription polymerase chain reaction (RT-PCR) to test
for the SARS coronavirus. In RT-PCR RNA is reverse transcribed into
its DNA complement and detected by binding it to a primer specific to
the human SARS coronavirus. In addition, the scientists also tested for
presence of the virus by inoculating the samples into cell cultures. If the
virus was present, the cells displayed degenerative changes associated with
the multiplication of viruses.
The beaver (Castor fiber), Chinese ferret-badger (Melogale moschata),
Chinese hare (Lepus sinensis), deer (Chinese muntjac, Muntiacus reevesi),
domestic cat (Felis catus) and hog-badger (Arctonyx collaris) never tested
positive in 18 total samples. However, all of the Himalayan palm civet
(Paguma larvata), raccoon dog (Nyctereutes procyonoides), 7 out of 7 ani
mals, tested positive. The Himalayan palm civet is a small carnivore found
throughout Southeast Asia and the raccoon-dog is a type of dog found in
eastern Asia.
Guan and his colleagues also collected blood samples from 20 wild
animal traders, 15 butchers, and 20 vegetable retailers. They tested these
samples for antibodies to the virus isolated from one of the palm civ
ets (Paguma larvata) using an immunofluorescence assay. Prior to being
tested, none of these individuals reported SARS-like symptoms. Higher
percentages of wild animal traders and butchers, individuals handling an
imal products, tested positive for the SARS coronavirus antibody (40%
and 20%, respectively) than vegetable retailers (5%).
Although Guan and his colleagues could not conclude that either the
civet or the raccoon-dog is the reservoir for the SARS coronavirus, its
presence in these market animals suggests a connection between animals
46 EFFECTS OF GENETIC AND PATHOGENIC DISEASES ON CELLS
amino acid sequence of the resulting protein. Among the humans, twenty
or more of the spike gene nucleotides varied among the patients with
many altering the spike protein. This suggests that the jump from civet
to humans occurred further in the past, because the coronavirus DNA
sequences in humans contained many alterations not observed in the
coronavirus isolated from civets and raccoon-dogs.
Animal viruses isolated from one market should be more similar to
each other than the human viruses isolated from Hong Kong, Guangdong,
Canada, and Vietnam, but this was not the case. The viruses isolated from
humans from five geographically separate sites were very closely related
genetically. Even though the total number of mutations was greater, many
of the same mutations occurred in all human virus samples. This close
relationship among the coronavirus isolates from humans indicates that
the virus had spread very rapidly from China to as far away as Toronto,
Canada, after it mutated and jumped from civets, but faster than it
evolved major new differences. The scientists also concluded that the
Chinese live animal markets may have played a role in amplifying the
SARS coronavirus and transmitting this disease to humans, as suggested
by the evidence reviewed earlier.
Recall that a physician infected by a patient in Guangdong later trav
eled to Hong Kong. With a pathogen like the SARS coronavirus, which
has such effective dispersal abilities, SARS could quickly spread beyond
Guangdong province, which is exactly what it did. The World Health
Organization (WHO) keeps statistics on many diseases and epidemics
and compiled data on cases of SARS from China and beyond (Figure 8).
In addition to the cases shown, over 2,500 probable cases of SARS from
Beijing, China, are not shown because date of onset was not available to
WHO. The date of onset for the initial case in countries that had more
than ten cases of SARS is shown in the figure, along with the number
of cases that ultimately occurred in those countries. Several countries
had outbreak onset on the same date, and the number of cases shown is
pooled for those countries.
As global pandemics go, SARS was short-lived but widespread. A
pandemic is an epidemic occurring over a wide geographic area and af
fecting a high proportion of the population. Within just a few months of
its appearance in Guangdong Province, SARS spread to more than two
48 EFFECTS OF GENETIC AND PATHOGENIC DISEASES ON CELLS
1 = China, 5,328
2 = Hong Kong, 1,755
160 3 = Canada, United States,
Vietnam, 341
140 4 = Philippines, Taiwan,
120 Singapore, 598
numner of cases
100 Guangdong
Provice outbreaks
80 3
60 2 4
40 1
20
0
2
03
03
0
l
r
ov
ov
ec
ar
ar
ay
ay
Ju
Ap
Ja
Ju
Ja
Fe
M
N
11
18
3
20
24
20
14
1
22
13
30
date of onset
Figure 8 SARS cases by week of onset. Total number of cases shown
is 5,910 out of 8,096 total cases. Not all of Chinas 5,328 cases are
shown in the graph (see text). The time frame for the Guangdong
Province outbreaks described in text is shown.
Source: Data from World Health Organization. http://www.who.int/csr/sars/ country/
table2004_04_21/en/ index.html.
and eventually broke the chain of transmission. However, SARS has not
been eradicated, and it could reemerge.
The SARS outbreak illustrates how diseases spread. In the case of
SARS, the emergence of a disease transmitted quickly from one host to
another through close contact led to an epidemic in southern China and
then to a pandemic. A global response was necessary to curb the spread
of the disease. Epidemiologists worked quickly and collaboratively to
study this new disease, which was caused by a pathogen that did not pre
viously infect humans. Pathogens are part of ecological systems and they
can infect multiple hosts. Pathogens can evolve to switch to new hosts,
and this often leads to disease outbreaks. As ecological systems change,
opportunities for new interactions among species arise.
Bibliography
Dandekar AA, Perlman S: Immunopathogenesis of coronavirus infec
tions: implications for SARS, Nat Rev Immunol 5(12):917927, 2005.
Guan Y, Zheng BJ, He YQ, et al.: Isolation and characterization of viruses
related to the SARS coronavirus from animals in Southern China,
Science 302(5643):276278, 2003.
World Health Organization: Global alert and response (GAR): severe
acute respiratory syndrome (SARS), World Health Organization (web
site): http://www.who.int/csr/sars/en/. Accessed July 3, 2014.
Zhong NS, Zheng BJ, Li YM, et al.: Epidemiology and cause of severe
acute respiratory syndrome (SARS) in Guangdong, Peoples Republic
of China, in February, 2003, Lancet 362(9393):13531358, 2003.
Conclusion
The themes of internal environments, communication, structure and
function, and cells coming from preexisting cells have all been illus
trated in this chapter. The focus has been on diseases and their effects
at the cellular and organismal levels. Cell function can be disrupted by
altered DNA, proteins, and by pathogenic organisms. The disruption
affects internal environments or ability to maintain a constant internal
environment. Similarly, cell-to-cell communication may be disrupted or
manipulated by pathogens. A single population of a pathogenic organism
can interact with multiple host species, affecting their ability to interact
with other species. Pathogens can have complex life cycles and cause dis
eases to spread. The study of cells can provide insights into how diseases
affect multicellular organisms. The cell links other levels of the biological
hierarchyeffects at the molecular level are manifested at the cellular
level, which can then affect the entire organism.
Glossary
antibodies. Any of a large number of proteins produced by specialized B cells
after stimulation by an antigen and acting against the antigen in an immune
response, and that typically consist of four subunits.
blast. Term used to describe a syndrome of plant diseases where afflicted leaves
suddenly die.
cells. The smallest structural and functional unit of an organism.
chromatography. A process in which chemicals carried by a liquid or gas are
separated into components as a result of differential distribution of solutes as they
flow over a stationary liquid or solid substrate.
cytokines. Cytokines are small proteins, peptides, or glycoproteins released by
cells involved in cell-cell interaction and communication.
denatured. To modify the molecular structure of a protein or DNA by heat, acid,
alkali, or ultraviolet radiation, and thus destroy or diminish biological activity.
disease. A disease is a condition of an organism that impairs normal functioning,
manifested by distinguishing signs and symptoms.
dispersal. Dispersal means to spread from the place of birth.
ectothermic. Of or relating to an animal that cannot regulate its own body tem
perature, so its body temperature fluctuates according to its surroundings.
electrophoresis. The process in which large molecules can be separated according
to size and electrical charge by applying an electric current to them in a gel.
emerging disease. Emerging diseases are infectious diseases that are new in a
population or are rapidly increasing in incidence or geographic range.
endothermic. Endotherms are animals that use metabolic processes to maintain
a relatively constant body temperature.
eukaryote. An organism whose cell or cells contains a distinct, membrane-bound
nucleus.
exon. Portions of eukaryotic mRNA that are retained during mRNA splicing.
familial ALS. An inherited form of amyotrophic lateral sclerosis (ALS), an incur
able and common form of motor neuron disease, a progressive neurodegenerative
disease that affects nerve cells in the brain and spinal cord.
hemoglobin. Hemoglobin is a protein within red blood cells that carries oxygen
and releases it to cells lacking oxygen.
immunofluorescence. The labeling of antibodies or antigens with fluorescent
dyes for the purpose of demonstrating their presence.
leucocytes. Leucocytes are white blood cells that help the body fight infections
and other diseases.
54 GLOSSARY
life cycles. The stages through which an organism passes from the beginning of
its life until its death.
motor neuron disease. A progressive and often fatal neurodegenerative disease.
motor neurons. Motor neurons convey electrical impulses from the nervous sys
tem to a muscle or gland.
mutations. Mutations are changes in a DNA sequence that may or may not alter
the phenotype.
normal. Conforming to a type or a standard, for instance, of a behavior.
osmotic pressure. Osmotic pressure is the pressure exerted by water through
a semi-permeable membrane separating two solutions with different solute
concentrations.
outbreak. An outbreak is an occurrence of disease greater than would otherwise
be expected in a particular time and place.
pandemics. A global outbreak, or epidemic.
pathogenic. Disease causing.
phagocytic. Describing the function of white blood cells that ingest microbes,
other cells, and foreign particles.
phylum. Phylum is the term used to denote the primary division of a kingdom,
ranked above class in size.
plasmolysis. Plasmolysis is the bursting of a cell due to influx of a large abun
dance of molecules.
pneumonia. A disease that affects the lungs and makes it difficult to breathe.
point mutation. A point mutation is a change in a single nucleotide in a DNA
sequence.
population. A population is a group of individuals of the same species living in
the same place at the same time
proteins. Macromolecules with have shapes that determine their function and are
formed by the assembly amino acids.
reservoir. An epidemiological reservoir is a species that possesses a pathogen and
can pass it to other species.
RNA. Chemically similar to DNA except the ribose retains the 2 oxygen.
ssRNA. single-stranded RNA
tissues. Collections of more than one cell type of a single species that work to
gether to perform a complex function.
trypsin. An enzyme that breaks down proteins.
vector-borne disease. Vector-borne diseases are diseases in which the pathogen is
transmitted to an individual by some other agent.
Index
Allison, Anthony, 78 Fused in sarcoma/translated in
Amyotrophic lateral sclerosis (ALS), 9 liposarcoma (FUS/TLS) gene,
mutations that co-inherited 910
with, 10 mutations affect function of, 11
Anemia, 7 reason for MND, 12
Animals in research, 2224
Arginine, 5 Genetic diseases
ethical, legal, social implications,
Bacteria both, 15 1214
Bacterial plasmid, 19 sickle-cell disease
Barbour, Alan, 36 by multiple mutations, 912
Black, Robert, 29 by single point mutation, 18
Blast, 2425 Gerbils, male, experiment on, 35
Borrelia burgdorferi, 1517 Giardia lamblia, 29, 30, 41, 44
culture of genetically engineered, 19 campers to live off land, 3638
hosts immune system, 22 in children, 3033
cysts in feces, 33
Campbell, J. D., 35 experiment on male gerbils, 35
Castor canadensis, 37 Glutamic acid, 5
Cells burst (plasmolyze), 26 Grimm, Dorothee, 16
Centers for Disease Control and Guan, Y., 41
Prevention (CDC), 29
Chemokines, 18 Hemoglobin, 2
Coronaviruses, 41 in carbon monoxide (CO), 2
animal, 4546 fingerprints, 45
human, 46 Himalayan palm civet, 45
severe acute respiratory syndrome. Host cells, 2728
See Severe acute respiratory Howard, Richard, 25
syndrome (SARS) Human coronavirus, 46
Craft, J. Carl, 34 Human fecal, infectivity of, 3435
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