Epidemiology Microbiology-242N

Time to put on your epidemiology hat! Investigate and describe one disease outbreak (epidemic, endemic, sporadic, pandemic, etc.) in the last 200 years. Explain why the outbreak was classified as such. As an epidemiologist, how would you prevent this type of outbreak in the future?

For the first post make sure to start with the reservoirs of diseases and your understanding in details and then describe the epidemiology with the important terms and what they mean in terms that we learned in lecture for chap 13.

Chapter 13

Microbe–Human Interactions

Lecture Outline

See separate PowerPoint slides for all figures and tables pre-inserted into PowerPoint without notes.

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1

Colonization, Infection, Disease

Infection: A condition in which pathogenic microorganisms penetrate host defenses, enter the tissues, and multiply

Pathologic state: Cumulative effects of infection damage, disruption of tissues and organs, results in disease

Disease: Any deviation from health. Factors –Infections, Diet, Genetics, Aging.

Infectious disease: Disruption of tissues or organs caused by microbes or their products.

Exogenous: originating from outside body- environment, another person, or animal

Endogenous: already existing on or in the body-Normal biota or a previously silent infection

 

 

 

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Importance of Gut Biota-Differences in gut microbiome have been associated with risk for: Crohn’s disease, Obesity, Heart disease, Asthma, Autism, Diabetes, Moods

Include an array of bacteria, fungi, protozoa, and viruses

have a profound effect on human biology

Human cells contain 22,000 protein encoding genes; microbes that inhabit humans contain 8 million (HUMAN MICROBIOME PROJECT)

 

Preliminary results:

Human cells contain 22,000 protein encoding genes; microbes that inhabit humans contain 8 million

We have a lot of microbes in places we used to think were sterile

All healthy people harbor potentially dangerous pathogens, but in low numbers

The makeup of your intestinal biota can influence many facets of your overall health

Differences in the gut microbiome have preliminarily been associated with differences in the risk for:

Crohn’s disease

Obesity

Heart disease

Asthma

Autism

Diabetes

Moods

 

 

Disease:

Any deviation from health

Factors that cause disease:

Infections

Diet

Genetics

Aging

 

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Relationships Among Resident, Transient, and Disease-Causing Microbes, and Human Host

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Resident Biota: Normal biota/ resident or indigenous biota or normal flora Large and diverse collection of microbes living on and in the body. Include an array of bacteria, fungi, protozoa, and viruses

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Acquiring Resident Biota

Benefits of normal biota: Prevent the overgrowth of harmful microorganisms. Development/ sustain immune system

Microbial antagonism: antagonistic effect “good” microbes have against intruder microorganisms

Skin and adjacent mucous membranes, Upper respiratory tract, Gastrointestinal tract, including mouth, Outer portion of urethra, External genitalia, Vagina, External ear canal, External eye (lids, conjunctiva)

Endogenous/Opportunistic Infections: Caused by biota already in the body.

Can occur when normal biota is introduced to a site that was previously sterile.

Weak Immune system-Age: very young/old, Genetic defects in immunity and acquired defects in immunity (AIDS), Pregnancy, Surgery and organ transplant, Chemotherapy/ immunosuppressive drugs , Physical and mental stress

 

Broad spectrum antibiotics

 

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Opportunistic pathogens: Cause disease when:

The host’s defenses are compromised

When they become established in a part of the body that is not natural to them

Endogenous infections: Caused by biota already in the body

Can occur when normal biota is introduced to a site that was previously sterile

Example: Escherichia coli entering the bladder, resulting in a UTI

 

4

Continued Colonization of the Baby Through Diet

Fetuses are seeded with normal microbiota in utero??

Exposure occurs during birth when the baby becomes colonized with the mother’s vaginal biota

Breast milk contains around 600 species of bacteria

Bottle feeding

Skin contact with people- staff, parents, grandparents

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A growing number of doctors and scientists believe fetuses are seeded with normal microbiota in utero

These microbes are important for healthy full-term pregnancies and healthy newborns

 

Sugars used by healthy gut bacteria

Breast milk may be necessary for maintaining a healthy gut microbiome in the baby

 

5

Will Disease Result From an Encounter Between a (Human) Host and a Microorganism?

Pathogen: A microbe whose relationship with its host is parasitic. Results in infection and disease.

True pathogens: Capable of causing disease in healthy persons with normal immune systems

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6

Primary Biosafety Levels and Agents of Disease

Biosafety Level Facilities and Practices Risk of Infection and Class of Pathogens
1 Standard, open bench, no special facilities needed; typical of most microbiology teaching labs; access may be restricted. Low infection hazard; microbes not generally considered pathogens and will not colonize the bodies of healthy persons; Micrococcus luteus, Bacillus megaterium, Lactobacillus, Saccharomyces.
2 At least level 1 facilities and practices; plus personnel must be trained in handling pathogens; lab coats and gloves required; safety cabinets may be needed; biohazard signs posted; access restricted. Agents with moderate potential to infect; class 2 pathogens can cause disease in healthy people but can be contained with proper facilities; most pathogens belong to class 2; includes Staphylococcus aureus, Escherichia coli, Salmonella spp., Corynebacterium diphtheriae; pathogenic helminths; hepatitis A, B, and rabies viruses; Cryptococcus and Blastomyces.
3 Minimum of level 2 facilities and practices; plus all manipulation performed in safety cabinets; lab designed with special containment features; only personnel with special clothing can enter; no unsterilized materials can leave the lab; personnel warned, monitored, and vaccinated against infection dangers. Agents can cause severe or lethal disease especially when inhaled; class 3 microbes include Mycobacterium tuberculosis, Francisella tularensis, Yersinia pestis, Brucella spp., Coxiella burnetii, Coccidioides immitis, and yellow fever, WEE, and HIV.
4 Minimum of level 3 facilities and practices; plus facilities must be isolated with very controlled access; clothing changes and showers required for all people entering and leaving; materials must be autoclaved or fumigated prior to entering and leaving lab. Agents are highly virulent microbes that pose extreme risk for morbidity and mortality when inhaled in droplet or aerosol form; most are exotic flaviviruses; arenaviruses, including Lassa fever virus; or filoviruses, including Ebola and Marburg viruses.

A system of biosafety categories adopted by the Centers for Disease Control and Prevention (CDC)-degree of pathogenicity , relative danger in handling these pathogens

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7

Virulence

Virulence: Degree of pathogenicity. Indicated by a microbe’s ability to: Establish itself in the host & Cause damage

Virulence factor: Any characteristic or structure of the microbe that contributes to toxin production or induction of an injurious host response.

Examples of Virulence factors

Slime layer- attachment to surface

Fimbriae- attachment to surface

Viral spikes- attachment

Capsules- hides from immune system (antiphagocytic)

Leukocidins- substances toxic to white blood cells (antiphagocytic)

Hemolysin- breaking down RBC

Coagulase- formation of clots of blood and plasma

 

 

 

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Infectious dose (ID): a minimum number of microbes required for an infection to proceed

Determined experimentally for many microbes

Microbes with a smaller infectious dose have greater virulence

Structures, products, or capabilities that allow a pathogen to cause infection in the host

Adaptations that a microbe uses to invade and establish itself in a host

 

 

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Steps Involved When a Microbe Causes Disease in a Host

Portal of entry: Exogenous / endogenous

Exogenous- A characteristic route taken by a microbe to initiate infection

Skin (Nicks, abrasions, punctures, exoenzyme)or mucous membranes- digestive (food/drink), respiratory (largest number of pathogens enter through nasal, oral), urogenital (STIs all genitals)

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Pathogens That Infect During Pregnancy and Birth

A few microbes cross placenta and are spread by umbilical vein into fetal tissues

TORCH: common infections of fetus and neonate

Toxoplasmosis

Other diseases: syphilis, coxsackievirus, varicella-zoster virus, AIDS, chlamydia

Rubella

Cytomegalovirus

Herpes simplex virus

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The placenta is an exchange organ:

Formed by maternal and fetal tissues

Separates the blood of the developing fetus from that of the mother

Permits diffusion of dissolved nutrients and gases to the fetus

A few microbes cross the placenta and are spread by the umbilical vein into the fetal tissues

Other infections are transmitted perinatally as the child passes through the birth canal

 

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Becoming Established: Step Two—Attaching to the Host

Adhesion:

binding between specific molecules on host and pathogen

Pathogen limited to only those cells/ organisms to which it can bind.

Klebsiella uses capsule

 

Viruses use spike to enter host cells

 

Escherichia coli, Neisseria gonorrhoeae- use fimbriae

Slime layer used to attach to surfaces

 

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Quorum sensing: Chemical communication between nearby bacteria critical to establishment of infection

 

Adhesion:

A process by which microbes gain a more stable foothold on host tissues

Dependent on binding between specific molecules on both the host and pathogen

A particular pathogen is limited to only those cells and organisms to which it can bind

Once attached, a pathogen can invade body compartments

 

11

Becoming Established: Step 3—Surviving Host Defenses

Phagocytes:

White blood cells that engulf and destroy pathogens by means of enzymes and antimicrobial chemicals- phagocytosis

Antiphagocytic factors:

Virulence factor used by pathogens to avoid phagocytes-capsules, Leukocidins

Circumvent some part of the phagocytic process

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12

Step Four: Causing Disease

Virulence factors:

Structures, products, or capabilities that allow a pathogen to cause infection in the host

Adaptations that a microbe uses to invade and establish itself in a host

Indirect damage-Determine the degree of tissue damage that occurs-interplay between microbe and host

 

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Many cases of microbial diseases are the result of indirect damage or the host’s excessive or inappropriate response to a microorganism

Pathogenicity is a trait not solely determined by microorganisms

Pathogenicity is a consequence of an interplay between microbe and host

 

13

Direct Damage via Enzymes

Exoenzymes:

Secreted by pathogenic bacteria, fungi, protozoa, and worms

Break down and inflict damage on tissues

Dissolve host’s defense barriers and promote the spread of microbes into deeper tissues

Examples:

Mucinase

Keratinase

Hyaluronidase

Hemolysin

Coagulase

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14

Origins and Effects of Circulating Exotoxins and Endotoxin

*A toxoid is an inactivated toxin used in vaccines.

**An antitoxin is an antibody that reacts specifically with a toxin.

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Toxin: A specific chemical product of microbes, plants, and some animals that is poisonous to other organisms

Exotoxin:

Secreted by a living bacterial cell to the infected tissues

Many types

Endotoxin: Lipopolysaccharide of outer membrane

Not actively secreted

Shed from the outer membrane

Only gram-negative bacteria

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15

Localized Infection

Microbe enters the body and remains confined to a specific tissue:

Boils, Fungal skin infections, Warts

Systemic Infection

When an infection spreads to several sites and tissue fluids, usually in the bloodstream, nerves or cerebrospinal fluid

 

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When an infection spreads to several sites and tissue fluids, usually in the bloodstream

Viral: measles, rubella, chicken pox, AIDS

Bacterial: brucellosis, anthrax, typhoid fever, syphilis

Fungal: histoplasmosis, cryptococcosis

 

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Focal Infection

Exists when the infectious agent breaks loose from a local infection and is carried to other tissues

Mixed Infection

Several agents establish themselves simultaneously at infection site

Polymicrobial diseases: gas gangrene, wound infections, dental caries, human bite infections

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Examples:

Tuberculosis

Streptococcal pharyngitis: scarlet fever

Toxemia: infection remains localized, toxins are carried through the blood to the target tissue

 

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Primary, Secondary Infections

Primary infection: Initial infection

Secondary infection: Occurs when a primary infection is complicated by another infection caused by a different microbe

Acute infections: Come on rapidly, have short-lived effects.

Chronic infections: Progress and persist over a long period of time

 

 

Acute versus Chronic Infections

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18

Signs and Symptoms: Warning Signals of Disease

Sign: Any objective evidence of disease as noted by an observer

More precise than symptoms-fever, chest sounds, Tachycardia, Abscesses, swollen lymph nodes, Septicemia, Microbes in tissue fluids, Skin eruptions, Leukocytosis Leukopenia

 

 

Symptom: Subjective evidence of disease as sensed by the patient.

Chills, Pain, ache, soreness, irritation, Malaise, Fatigue, Chest tightness, Itching , Headache, Nausea , Abdominal cramps, Anorexia (lack of appetite)

 

Syndrome: A disease identified or defined by a certain complex of signs and symptoms. AIDS

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19

Infections That Go Unnoticed

Asymptomatic, subclinical, or inapparent infections:

Host is infected but does not manifest the disease

Patient experiences no symptoms or disease and does not seek medical attention

Latency: A dormant state of an infectious agent

During this state, a microbe can periodically become active and produce a recurrent disease

Sequelae: Long-term or permanent damage to organs and tissues

Persistence of Microbes

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20

Vacating the Host: Step Five—Portals of Exit

Portal of exit:

Avenue for pathogens to exit the host

Secretion- coughing, sneezing

Excretion- urogenital tract, feces

Discharge

Sloughed tissue-skin.

Removal of blood

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Some intestinal pathogens cause irritation in the intestinal mucosa that increases the motility of the bowel

Resulting diarrhea provides a rapid exit for the pathogen

Helminth worms release eggs and cysts through the feces

Feces containing pathogens are a public health problem when allowed to contaminate drinking water or when used to fertilize crops

Respiratory and Salivary PortalsEscape media for pathogens that infect the upper and lower respiratory tract: Mucus, Sputum, Nasal drainage

The outer layer of skin and scalp is constantly being shed into the environment

Household dust is composed of skin cells

A single person can shed several billion skin cells a day

Blood has a portal of exit when it is removed or released through vascular puncture

Blood-feeding animals are common transmitters of pathogens:Ticks, Fleas, Mosquitoes

 

 

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Stages in the Course of Infection and Disease

Incubation period -time from initial contact with infectious agent to appearance of first symptoms.

Prodromal stage- notable symptoms start

Period of invasion- Infectious agent multiplies at high levels, exhibits its greatest virulence, and becomes well established in its target tissue

Convalescent- symptoms decline, recovery.

Continuation- only some infections, chronic Lyme disease, typhoid

 

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The incubation period is the time from initial contact with the infectious agent (at the portal of entry) to the appearance of the first symptoms. During the incubation period, the agent is multiplying at the portal of entry but has not yet caused enough damage to elicit symptoms. Although this period is relatively well defined and predictable for each microorganism, it does vary according to host resistance, degree of virulence, and distance between the target organ and the portal of entry (the farther apart, the longer the incubation period). Overall, an incubation period can range from several hours in pneumonic plague to several years in leprosy. The majority of infections, however, have incubation periods ranging between 2 and 30 days. The earliest notable symptoms of infection usually appear as a vague feeling of discomfort, such as head and muscle aches, fatigue, upset stomach, and general malaise. This short period (1 to 2 days) is known as the prodromal stage. Some diseases have very specific prodromal symptoms. Other diseases have an imperceptible prodromal phase. Next, the infectious agent enters a period of invasion, during which it multiplies at high levels, exhibits its greatest virulence, and becomes well established in its target tissue. This period is often marked by fever

 

22

Living Reservoirs

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Reservoir:

Primary habitat in the natural world from which a pathogen originates

 

Human or animal carrier; soil, water, or plants

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23

Animal reservoirs- Zoonoses

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An infection indigenous to animals but also transmissible to humans

Human is the dead-end host and does not contribute to the natural persistence of the microbe

Spread of disease is promoted by close associations of humans with animals

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24

Carrier States

Carrier:

An individual who inconspicuously shelters a pathogen (asymptomatic) and can spread it to others without knowing

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Vectors

In epidemiology, a live animal that transmits an infectious agent from one host to another

Majority of vectors are arthropods

Biological vector: Actively participates in a pathogen’s life cycle. Serves as a site in which the pathogen can multiply or complete its life cycle

Mechanical vectors: Not necessary to the life cycle of an infectious agent, Merely transport the pathogen without being infected

 

Biological vectors are infected

Mechanical vectors are not infected.

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Nonliving Reservoirs

Microbes have adapted to nearly every habitat in biosphere

Soil, water, and air

Most are saprobic and cause little harm to humans

Some are opportunists

A few are regular pathogens

© Christopher Kerrigan/McGraw-Hill Education

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27

Acquisition and Transmission of Infectious Agents

Communicable disease: infected host can transmit infectious agent to another host and establish infection in new host.

Contagious: agent is highly communicable, especially through direct contact.

Noncommunicable: Does not arise through transmission of the infectious agent from host to host.

Horizontal transmission: Disease is spread through a population from one infected individual to another.

Vertical transmission: Transmission from parent to offspring via ovum, sperm, placenta, or milk

 

 

 

 

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28

Patterns of Transmission in Communicable Diseases

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Indirect Spread

Vehicle: Any inanimate material commonly used by humans that can transmit infectious agents.

Fomite: An inanimate object that harbors and transmits pathogens.

Oral-fecal route: Fecal carrier with inadequate personal hygiene contaminates food during handling, unsuspecting person ingests it.

Water and soil : can be temporarily contaminated with pathogens that come from humans.

Air: Indoor air can serve as a support medium for the suspension and dispersal of respiratory pathogens via droplet nuclei (pellets of mucus and saliva are ejected from mouth nose)

and aerosols (Suspensions of fine dust/ moisture particles in air with live pathogens)

 

 

 

 

 

 

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Droplet nuclei:

Dried microscopic residues created when microscopic pellets of mucus and saliva are ejected from the mouth and nose

 

Fomite:

An inanimate object that harbors and transmits pathogens

Not a continuous source of infection

 

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Healthcare-Associated Infections

Infectious diseases that are acquired or develop during a hospital stay or stay in another health-care facility

Rates of HAIs can range from 0.1 to 20% of all admitted patients.

Medical asepsis: Practices that lower microbial load in patients, caregivers, hospital environment.

Surgical asepsis: Ensuring all surgical procedures are conducted under sterile conditions

 

 

*2014 data indicate these have started to decline.

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Infection control officer:

Implements proper practices and procedures throughout the hospital

Charged with:

Tracking potential outbreaks

Identifying breaches in asepsis

Training other health-care workers in aseptic technique

 

31

Using Koch’s Postulates to Determine Etiology

Essential aim of study of infection and disease is determining etiologic agent (causative agent)

Robert Koch:

Developed a standard for determining causation of disease that stood the test of scientific scrutiny

Determined the causative agent of anthrax

Koch’s postulates:

A series of proofs that established classic criteria for etiologic studies

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32

Epidemiology: Study of Disease in Populations

Epidemiology:

Effects of diseases on the community

Study of frequency and distribution of disease and distribution of disease and other health-related factors in defined populations

Reportable diseases: diseases considered to be of great public health importance- malaria, hepatitis, anthrax. (https://wwwn.cdc.gov/nndss/conditions/notifiable/2019/)

Notifiable diseases

By law, some diseases must be reported to authorities

Other diseases are reported on a voluntary basis

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33

Centers for Diseases Control and Prevention

Responsible for keeping track of infectious diseases nationwide

Part of the U.S. Public Health Service, incidence and prevalence can be tracked.

Disease trends and areas can be identified

High-risk epidemics can be identified and dealt with as quickly as possible.

The CDC shares its statistics on disease with the World Health Organization (WHO) for worldwide tabulation and control

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34

Epidemiological Statistics

Prevalence of disease: Total number of existing cases with respect to the entire population

Incidence of disease: Measures the number of new cases over a certain time period

Also known as case or morbidity rate

Mortality rate: Measures the number of deaths in a population due to a certain disease

Index case: The first patient found in an epidemiological investigation

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35

More Epidemiological Terms

Endemic: An infectious disease that exhibits a relatively steady frequency over a long time period in a particular geographic locale.

Epidemic: Whenever a disease occurs at a greater frequency than normal for a population or area

Sporadic disease: Occasional cases are reported at irregular intervals in random locales

Pandemic: Spread of an epidemic across continents

 

 

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Propagated epidemic:

Results from an infectious agent that is communicable from person to person and is sustained over time in a population

 

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Golgi Maturation

The way that proteins move through the golgi has long been a source of contention. Two major models have been proposed. One is a cisternal maturation model, and the other is a model of stable cisternae and mostly vesicular transport. In this paper they set out to try and differentiate these models using a modified Golgi resident protein.

 

In this paper they talk a bit about Golgi Tubules, these are like vesicles but form as long tubes rather then little circles… don’t worry too much about this issue.

 

 

Intro Questions (from last weeks panopto’s and the intro of the paper)

1) What is the main role of the golgi? How do the processes that are occurring differ from one stack to another (in other words, why have different stacks) – what is happening to proteins in these stacks?

2) What are the two main models of Golgi maturation? What is the different between Golgi resident proteins, and proteins moving through the endomembrane system? How would you expect the paths of these two protein types to be different?

 

Figure Questions:

 

Figure 1: What is Mani? How is Mani-FM different, and what is happening to Mani-FM when they add AP what about when AP is absent? What is the pellet vs supernatant test telling them?

 

Figure 2: What is Figure 2 showing them? Why did they do this test?

 

Figure 3: What are the main findings of figure 3?

 

Figure 4 a-i : What do they find about the localization of MANI-FM with and without AP within the golgi? Which of the two mechanisms of golgi maturation does this support?

 

Figure 4 j&k – what does this tell them about Mani-Fm with and without AP?

 

Figure 5: How is what they are doing in this figure different from what they did in Figure 4? What is this showing them? What did the washout tell them that the initial application did not? Given your knowledge of golgi trafficking, what kind of vesicles would you expect to fine Mani-FM in?

 

Conclusion Questions: What are the overall conclusions from the paper? Which model of Golgi trafficking does this support? Why is this interesting to us?

 

More Challenging questions: How are there findings different from previous work? What did they show that no-one has showed about this model before?

 

Be sure to include a bibliography of your sources.

 
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Name of enzyme you will use: Catalase

Created an outline.

 

Include the following in your outline:

 

Name of enzyme you will use

Name of organism (if applicable)

The substrate and products in the chemical reaction

Method for measuring enzyme activity

Treatment: acidic fluid(s), pH, length of exposure, how you will treat your samples

The control(s) in the experiment

Hypothesis

How you will present your data (table and/or type of graph)

Anything else you would like to get feedback on before you start your experiment

 

****Here is the outline*****

 

Outline to Final Applied Lab Project:

Enzymes Reaction to Hydrogen Peroxide and Temperatures

 

Name of enzyme you will use:  Catalase

 

Substrate and products in the chemical reaction:  Hydrogen Peroxide

 

Method for measuring enzyme activity:  Balloons

 

Controls:  Independent: Temperature

Dependent:  Balloon

 

Length of exposure:  10 minutes

 

Hypothesis:  The response of enzyme to Hydrogen Peroxide and a specific        temperature

 

How you will present your data: Bar Graph

 

Summary of findings

 

*************************************

 

 

 

Instructions:

 

Demonstration of your knowledge of basic laboratory skills, experimental design, and/or data evaluation will be assessed by the submission of an applied final lab project.

 

Based on the outline, and the Lab submitted, the 4 outcomes need to be addressed

Laboratory Assignments

Addresses course outcomes 1-4:

·       recognize and explain how the scientific method is used to solve problems

·       make observations and discriminate between scientific and pseudoscientific explanations

·       weigh evidence and make decisions based on strengths and limitations of scientific knowledge and the scientific method

·       use knowledge of biological principles, the scientific method, and appropriate technologies to ask relevant questions, develop hypotheses, design and conduct experiments, interpret results, and draw conclusions

 

 

 

Experiment 2: Effect of Temperature on Enzyme Activity

Pre-Lab Questions

1. What reaction is being used to study the effect of temperature on enzyme activity in this experiment? Temperature can affect the enzyme catalytic reaction.

 

2. What does an increase in balloon diameter mean with respect to enzyme activity? The activity increases as well

 

 

3. Why is important to know this information before starting Experiment #1? The diameter of the balloon will show the activity of the enzyme based upon temperature.

 

Table 2: Balloon Circumference vs. Temperature
Tube Temperature (°C) Uninflated Balloon Circumference (cm) Final Balloon Circumference (cm) Difference in Balloon Circumference (cm)
1 – (Cold) 1  4 inches  6.5 2.5
2 – (RT)  2.3    6.75 2.75
3 – (Hot)  4    7.25 3.25

 

Insert labeled photos of your initial and final balloons here. Photos must be labeled! Include a note with your name and date on an index card in the pictures.

 

Post-Lab Questions

 

1. What is the enzyme in this experiment? The catalase. What is the substrate? The Hydrogen Peroxide

 

2. What is the independent variable in this experiment? Temperature. What is the dependent variable? Balloon diameter

 

3. How did temperature affect enzyme function? Hot temperature causes high enzyme activity, the cold has less activity, but the room temperature’s activity was slower to react.

 

 

4. Is there a negative control in this experiment? No negative controls. To revise we would use water, therefore nothing would be expected to happen.

 

 

 

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Experiment 4 Exercise 1 – Mitosis in a Plant Cell

How to Proceed

  • Read through the introductory materials below.
  • Open the Unit 4 Experiment Answer Sheet and complete the following Experiment exercises this unit:
    • Experiment 4 Exercise 1 – Mitosis in a Plant Cell (~45 min)
    • Experiment 4 Exercise 2 – Meiosis (~30 min)
    • Experiment 4 Exercise 3 – Karyotyping (~60 min)
  • Save your completed Unit 4 Experiment Answer Sheet and submit it no later than Sunday midnight (CT).

Mitosis – Introduction

The life cycle of a typical cell is roughly divided into two phases, Interphase and Cell Division. Interphase is a time of growth and synthesis and is further divided in to G1, S and G2 stages. G stands for “growth”. During G1, the cell grows and makes materials needed to replicate its DNA. DNA is then replicated during the S or “synthesis” phase. Additional growth occurs during G2 as the cell prepares for cell division. Cell division encompasses both nuclear division (Mitosis) and cytoplasm division (Cytokinesis). See Fig 8.6 on p 125 in your book.

Mitosis allows cells in our body to regenerate (e.g., replacement of shed skin cells) and to repair damaged tissue, since mitosis produces daughter cells that are identical to the parent cell. Before beginning this exercise, you will want to review Mitosis in our online lecture and examine the diagrams in Fig 8.7 on pp 126-127 and Fig 8.8 on p 128, which illustrates some differences between animal and plant cells. As you can see, each phase of the cell cycle can be recognized based on the appearance the chromosomes and nucleus. In this first exercise, you will need to be able to identify the various phases of the cell in onion cells. Below is an example of what you will be looking at:

Phases of the cell in onion cells

From: Ackroyd, Miller and Sauriol. No date. Mitotic cell division in onion root tip, Allium cepa.

Slide # Phase Description
1-2 Interphase Chromosomes are not visible in the nucleus (gray circle).
3-5 Prophase Chromosomes have become visible; although not apparent, the nuclear membrane is breaking down.
6 Metaphase Chromosomes have aligned along the equator (mid-line).
7-8 Anaphase Sister chromatids have separated and are being pulled to opposite poles.
9-10 Telophase Chromosomes are clumped at opposite poles, the nucleus has reformed and a cell plate is forming.

Note that is hard to determine exactly when one phase ends and the next one starts. Slide #7, for example is early anaphase and could also be considered late metaphase. Slide #9 is early telophase, but might also be considered late anaphase.

See your Unit 4 Experiment Answer Sheet for the instructions and the photographs you will use to complete this exercise.


Meiosis – Introduction

Meiosis is the process by which gametes (eggs and sperm) are produced from a germ cell. This type of cell division is unlike mitosis in that the cells produced are genetically different than the parent cell and the resulting cells contain only half the number of chromosomes (haploid). In this exercise, you will need to demonstrate your understanding of Meiosis and the role it plays in sexually reproducing organisms. Review this unit’s reading in your book (pp 131-141) and this unit’s online lecture before completing this exercise. You will need to review the following animation, so be sure that you are able to open and view it:

McGraw-Hill Higher Education. 2006. How Meiosis Works
http://highered.mcgraw-hill.com/sites/0072495855/student_view0/chapter28/animation__how_meiosis_works.html (Links to an external site.)

When you are ready, open the Unit 4 Experiment Answer Sheet and answer the questions for this exercise.


Karyotyping – Introduction

Review pp 130-121, pp 138-140 and our online lecture this unit before beginning this exercise. Many genetic disorders arise as a result of errors during Meiosis and the formation of gametes (= egg and sperm). Frequently, these errors are due to non-disjunction, in which chromosomes fail to separate correctly during Anaphase. When this happens, a daughter cell ends up with either an extra chromosome or it is missing a chromosome. These genetic disorders can be detecting using a screening process called a karyotype. In this exercise, you will learn how a karyotype is generated and used to diagnose the presence or absence of a disorder.

You will use the following website to complete this exercise:

The Biology Project. 1998. Karyotyping Activity
http://www.biology.arizona.edu/human_bio/activities/karyotyping/karyotyping.html  V

WEEK 4 EXPERIMENT ANSWER SHEET Please submit to the Week 4 Experiment dropbox no later than Sunday midnight.

SUMMARY OF ACTIVITIES FOR WEEK 4 EXPERIMENT ASSIGNMENT

· Experiment 4 Exercise 1 –Mitosis in a Plant Cell

· Experiment 4 Exercise 2 – Meiosis

· Experiment 4 Exercise 3 – Karyotyping

Experiment 4 Exercise 1: Mitosis in a Plant Cell

Read through the Experiment 4 Introduction material before starting. In this exercise we will look at the different stages of mitosis in onion cells. The length of the cell cycle in the onion root tip is about 24 hours, but mitosis only occupies only one to two hours.

Procedure

A. Review the information on p 125 in your book and record your predictions as to the percentage of cells you expect to see in each of the phases of the cell cycle in Table 1 (after the photographs).

B. In the four photographs below, examine each cell indicated by a red dot. There are a total of 65 cells that need to be examined. Determine how many cells are in the various stages of the Cell Cycle. Refer to your book and the Experiment 4 Introduction information for help determining the cell cycle stages.

C. Enter your data in Table 1. Note that the “Calculated %” is equal to the Number of cells in stage / Total cells counted x 100.

D. Answer the questions that follow.

image1.png

image2.png

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Table 1: Predictions and actual number of cells observed in each stage of the cell cycle (2 pts)

Stage Predicted % Number of Cells Total Cells Counted Calculated %
Interphase     65  
Prophase     65  
Metaphase     65  
Anaphase     65  
Telophase     65  
Total 100% 65 65 100%

Questions

1. What stage of the cell cycle were most of the onion root tip cells in (1 pts)?

Is this what you would expect based on what you’ve read this week; why or why not? Cite source(s) used (1 pts).

2. How accurate were your predictions, based on your data, for each stage of the cell cycle (1 pts)?

3. What is the function of mitosis in an organism such as a human? Cite source(s) used (2 pts).

Experiment 4 Exercise 2: Meiosis

Review this week’s reading in your book (pp 131-141) and this week’s online lecture. Then view the following animation before answering the questions below; be sure your audio is on:

McGraw-Hill Higher Education. 2006. How Meiosis Works http://highered.mcgraw-hill.com/sites/0072495855/student_view0/chapter28/animation__how_meiosis_works.html

Questions

1. Why is it necessary to reduce the chromosome number in gamete cells (egg and sperm), but not other cells of an organism? Cite source(s) used (2 pts).

2. Describe at least two differences between Meiosis I and Meiosis II in terms of what is produced and what occurs with the chromosomes. Cite source(s) used (2 pts).

3. If humans have 46 chromosomes in each of their body cells, determine how many chromosomes you would expect to find in the following cells (4 pts):

Sperm _________

Liver cell _________

Daughter cell from mitosis _________

Daughter cell from Meiosis II _________

4. _____ are separated during Meiosis I, while _____ are separated during Meiosis II (2 pts).

a. Sister chromatids, sex chromosomes

b. Sister chromatids, homologous chromosomes

c. Homologous chromosomes, sister chromatids

d. Sex chromosomes, sister chromatids

5. At the beginning of meiosis I, cells are ____ and at the beginning of meiosis II, they are ____ (2 pts).

a. Diploid, diploid

b. Diploid, haploid

c. Haploid, haploid

d. Haploid, diploid

Experiment 4 Exercise 3: Karyotyping

Go to the following website:

The Biology Project. 1998. Karyotyping Activity

http://www.biology.arizona.edu/human_bio/activities/karyotyping/karyotyping.html

Procedure

A. Read over the material on the first page and then click on Patient Histories (at the bottom of the page).

B. Click on Complete Patient A’s Karyotype. Note a single chromosome located to the left of the paragraph at the top. You need to match this chromosome with the appropriate pair.

a. Some of the chromosomes arranged beneath the paragraph are hyperlinked (have a blue line beneath them).

b. Click on the hyperlink associated with the matching chromosome.

C. Record in Table 2 below the correct match for this first chromosome (a).

D. Continue until you have matched all chromosomes required. Note that the letters (a-e) correspond to the order that the chromosomes are presented.

Table 2. Patient A karyotype data (1 pts).

Chromosome Presented Matches #
Chromosome a  
Chromosome b  
Chromosome c  
Chromosome d  
Chromosome e  

Questions

1. What notation would you use to characterize Patient A’s karyotype? Be sure to read over the information presented so you know the correct notation format (1 pts).

2. Based on the table of possible chromosomal disorders displayed on the website, what diagnosis would you give patient A (1 pts)?

3. When during Meiosis does the error occur that results in this karyotype; be specific (1 pts)?

 

Procedure (continued)

E. Click on the Patient History button (bottom of the page) and then click on Complete Patient B’s Karyotype.

F. Match the chromosomes presented as done in the previous exercise.

G. Fill in the Table 3 below, indicating how you matched up the chromosomes presented with the actual chromosomes in the karyotype.

Table 3. Patient B karyotype data (1 pts).

Chromosome Presented Matches #
Chromosome a  
Chromosome b  
Chromosome c  
Chromosome d  
Chromosome e  
Chromosome f  
Chromosome g  
Chromosome h (This is a tough one)  

Questions

4. What notation would you use to characterize Patient B’s karyotype (1 pts)?

5. What diagnosis would you give patient B (1 pts)?

6. Why does this individual exhibit male characteristics and not female? Cite your source (2 pts).

 

Procedure (continued)

H. Click on the Patient History button (at the bottom) and then click on Complete Patient C’s Karyotype.

I. Match the chromosomes presented as done in the previous exercise.

J. Fill in Table 4 below, indicating how you matched up the chromosomes presented with the actual chromosomes in the karyotype.

Table 4. Patient C karyotype data (2 pts).

Chromosome Presented Matches #
Chromosome a  
Chromosome b  
Chromosome c  
Chromosome d  
Chromosome e  
Chromosome f  
Chromosome g  
Chromosome h  
Chromosome i  

Questions

7. What notation would you use to characterize Patient B’s karyotype (1 pts)?

8. What diagnosis would you give patient C (1 pts)?

9. Why might the risk of this disorder increase as a woman gets older? Cite any sources used (3 pts).

 

Week 4 Experiment Grading Rubric

Component Expectation Points
Experiment 4 Exercise 1 Distinguishes among the various stages of the cell cycle and correctly answers the questions (Table 1, Questions 1-3). 7 pts
Experiment 4 Exercise 2 Demonstrates an understanding of meiosis and how it relates to sexual reproduction (Questions 1-6). 12 pts
Experiment 4 Exercise 3 Creates and uses a karyotype to diagnose genetic disorders (Tables 2, 3, and 4; Questions 1-9). 16 pts
TOTAL  
 
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