An Inquiry into the Effects of Tobacco Smoke on the Lungs Using a Hooka

To begin with, the experiment that was conducted in the laboratory was on the effect of cigarette smoke on mollusc gill cilia. The cigarette smoke is composed of toxins, and oxidative chemicals that poses a major stress on the airway epithelium. “Each puff of cigarette smoke contains >1014 oxidant molecules and >1000 xenobiotics , and exposure to cigarette smoke evokes significant biologic changes in the airway epithelium, even though many smokers are phenotypically normal” (Leopald, 2009). Smokers are subjected to about 4,000 toxic chemicals in cigarette smoke, including arsenic, methane, and carbon monoxide. The continuation of smoke exposure on the lungs effect the cilia because the dirt, environmental pollutants, and toxins from cigarette smoke accumulate in the cilia causing the toxins to remain in the lungs. Furthermore, these toxins migrate from the lungs by the means of blood transportation to other organs. The smoke from all cigarettes is not necessarily equivalent, with the amounts of toxic substances varying from one brand to another. It is possible to specify the amount of tar, nicotine, and carbon monoxide that is delivered by a cigarette under experimental conditions, which is what we will be observing in this experiment. The source of the tobacco, the length of the cigarette, the type of paper and filter used, how densely the tobacco is packed, the temperature at which it burns, and the rate at which the smoke is drawn are just some of the factors that will influence what reaches an individuals respiratory system.
Cancers of the lung are mainly the result of the poisonous exposure of the cigarette smoke to the lungs. The purpose of this experiment is to examine the effects of the compounds in cigarette smoke on the active cilia and to test whether varying particular parameters associated with smoking will influence the magnitude of the effect. We will do this by making an aqueous extract of smoke, by drawing it through a hooka by using the cilia from a mollusc gill. We will be studying the effects on the beating of cilia from seawater, and from a filtered and non filtered cigarette. The brand of cigarette that will be used is Export A (Heavy) which contains roughly 15-33 mg of tar; 12-26 mg of nicotine; and 14-28 mg of carbon monoxide. Our hypothesis states that the contamination will lead to faster cilia beating. The rationale behind the hypothesis is that the more the water is contaminated from the different types of treatment in the hooka the faster the beating of the cilia will be because the harmful effects of smoking will negatively effect the cilia and cause them to beat at an abnormally fast rate which will in turn result in negative impacts on the body and respiratory system. My prediction is that the non filtered cigarettes will be more damaging to the respiratory system and therefore more tar and other residuals will be formed on the hooka, which is to mock the respiratory system.

Methods
An Export A (heavy) cigarette is taken and will be used to examine the effects on the cilia. Smoke the cigarette in a hooka, to do this you must insert the cigarette into tubing. Squeeze the bulb and force air out, then immediately put your thumb over the open end while the bulb is still held squeezed. Hold a lit match to the cigarette, and with your thumb over bulb opening realse pressure on bulb so air is drawn through the cigarette opening into the hooka and into the bulb. Release thumb from end of bulb and squeeze the smoke out. The rate at which the hooka will be smoked is 10 puffs per minute and the whole cigarette smoked. This will be done all in the fumehood.
The experiment will be repeated again this time however using an Export A (heavy) cigarette with no filter. The appearances of the filtered and non filtered cigarette are then recorded.
Obtain a small section, about 1 x 5 mm, that has been cut from a mollusk gill and mount it in saline solution on a microscope slide. Leave the tissue to recover for approximately five minutes.
Under the lowest power of the microscope observe the action of the cilia and the speed of the cilia in beats per minute.
Obtain a fresh sample of mollusc gill. Follow the same procedure as for the control experiment, except using water from the hooka of the filtered and non filtered cigarettes instead of clean saline.
Observe the cilia in hooka water under the microscope and compare their action with cilia of the control cells. After letting the tissue recover, count the number of beats per minute as before.
All the results will be recorded from each student, which will be later used to examine the impacts of smoking on the cilia.
Afterwards, obtain a microscope slide with a section of a normal mammalian lung. Find amongst the alveoli, a cross section of a lung bronchus.
Under the microscope (low power), it will look like a convoluted tube.
Focus at high power (40X) on the epithelial layer of the bronchus. Once you have located draw and label a high power view of the bronchial epithelium. Label the ciliated columnar, epithelial cells and nucleus, mucous layer, bascal cells and goblet cells. Obtain a microscope slide with a section of bronchiogenic carcinoma of the lung. Black masses in the tissues will be present. Draw a typical region from the cancerous tissue.

Results:
Hooka Description:
The filtered and non filtered cigarette was smoked fully through a hooka, which symbolizes the respiratory track was conducted to show the different effects of each cigarette type. The results showed that the filtered cigarette beaker and tube were cleaner than that of the non filtered cigarette. The filtered hooka had a lighter yellow residual substance that was formed on the tube, this makes sense because filters have the ability to capture and retain detrimental toxins from the cigarette such as nicotine and tar. In comparison, the non filtered hooka there is an excessive amount of build up resulting in a yellow brownish red colour in the tube. The increase in pollutants shown on the non filtered hooka indicate that the effects of the non filtered cigarettes are more harmful to the cilia and the respiratory system.

Graph:

In the graph comparing the cilia beating of seawater, filtered and non filtered cigarettes, the seawater treatment resulted in the highest cilia beating, afterwards the non filtered cigarette following by the filtered cigarette. The vertical line on the top of each bar indicates the standard deviation for each set of data recorded.

Discussion
Cilia are small found on the surface of cells of a wide variety of organisms. Single celled organisms may propel themselves through water with cilia, and multicellular organsims use the beat of cilia to move material along. Cilia are tiny hair-like organelles that reside on the surface of cells for the purpose of sweeping debris away from lung and nasal cavities. The cilia are in constant motion, sweeping the mucus and particles mixed in it toward the throat where they are swallowed. The epithelial lining of our respiratory tract is covered with cilia (Sisson, 1994). The cilia are constantly beating, movie dust, particles and dead cells trapped in mucus out of our lungs. The mucous traps the little bits of dirt and pollution you breathe in, and the cilia move together like a wave to push the dirt-filled mucous out of your lungs. Then you cough, swallow, or spit up the mucous, and the dirt is out of your lungs. The most common reason to suffer damaged or paralyzed cilia, however, is smoking. Normally, healthy cilia in the bronchial tubes work with specialized cells that produce mucous to capture and remove impurities out of the lungs (Campbell, 2000). There is a thin layer of mucous and thousands of these cilia lining the insides of your breathing tubes. “One of the consequences of smoking is the associated reduction in mucociliary clearance, the process by which the coordinated action of cilia on the airway epithelium moves the airway surface fluid and mucus in a cephalad fashion, providing continuous cleansing of the airway surface. In the context that mucociliary clearance is a vital defense against inhaled pathogens and particulates, a reduction in mucociliary clearance contributes to the increased susceptibility of cigarette smokers to respiratory tract infection, and to the increased risk for the development of chronic obstructive lung disease and bronchogenic carcinoma” (Stanley, 1986). As the cigarette smoke moves through the bronchi, mucus layer tends to trap the cancer-producing tars along with other particles. In response to the presence of excess irritants, mucous cells in the lungs produce more mucous than they normally would. In Figure 2 in the appendix the sections of the cancerous lung that we examined, darker spots are seen. These are likely deposits of tar from cigarette smoking. Of course, with damaged cilia, the lungs have no means of moving the mucous out. These harmful agents cause mutations, chromosomal aberrations and other damage. In the experiment the seawater is the control variable because it contain an excessive amount of harmful pollutants which is why the seawater is 219.65 beats per minute as compared to that of a filtered cigarette which is composed of volatile toxins is 178.25 beats per minute. When your lungs' natural cleaning and repair system is damaged, germs, dirt and chemicals from cigarette smoke stay inside your lungs. This puts you at risk for bad cough and chest infections. With the cilia cells destroyed they become displaced by an excessive number of basal cells. Over time, these cells may become cancerous. They may also become extremely unsafe, escaping through lung tissue and invading other parts of the body. Nicotine and tar quantities were 25% and 30% less, respectively, for filter cigarettes than with non filtered cigarettes. In theory, the purpose of the filter is to reduce the amount of tar and nicotine that a smoker inhales with each breath through a series of holes that combine fresh air with the cigarette smoke, making the cigarettes ‘safer’. (Lungerella, 2007). This does not correspond to our results in the experiment because the non filtered cigarette is 185 beats per minute while the filtered cigarette has a beating of 178 beats per minute. We know that these values are inaccurate and therefore inconclusive because the non filtered cigarette should have a slower beating as more tar and other toxins are entered into the respiratory system. The filters reduced the amount of tar and nicotine inhaled when smoking the filtered cigarette as compared to the non-filtered cigarette. On major flaw that anti-tobacco enthusiasts find in this testing method is that each human being smokes differently than a programmed machine.

References:
Campbell, Neil. Reece, Jane. 2002. Biology: Sixth Edition. Pearson Education, Inc. San Francisco, CA
Leopold PL, O'Mahony MJ, Lian XJ, Tilley AE, Harvey B-G, et al. (2009) Smoking Is Associated with Shortened Airway Cilia. PLoS ONE 4(12): e8157. doi:10.1371/journal.pone.0008157

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