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Hi, I need someone to help me turn my rough cell biology draft into the final draft. Down below I will update both lab experiments I performed to complete this paper. Both my experiment proposals. I will upload my draft and someone to explain my tables in my draft. Fix the fonts- and add page numbers- follow the rubric and make sure I achieve a high score Please and thank you. FYI: image 9317 and 9318 are the only tables neccsariy.
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BIOS 312 Cell Biology Lab Research Project: Effects of E-cigarette Liquid on
Tetrahymena Phagocytosis
Student Name
Course and Semester: BIOS 312 – Fall 20XX
Introduction
Background Information
Due to health concerns, the global surge in e-cigarette use has piqued scientific interest.
The early 2000s Chinese invention of e-cigarettes (Schraufnagel et al., 2014) has become
popular worldwide, especially in China, the US, and Europe. E-cigarettes, marketed as a safer
alternative to tobacco cigarettes, vaporize e-liquids or “juice,” which contain various ingredients.
Most e-liquid ingredients are “generally recognized as safe” (GRAS) by the FDA, however
aerosol inhalation is controversial.
Nicotine, propylene glycol, glycerol, and flavorings can be found in e-liquids (Blount et
al., 2020). E-liquids, especially handmade ones, may include THC and vitamin E acetate. These
additives may harm health. Studies have shown that inhaling flavorings like diacetyl can cause
respiratory tract damage like “popcorn lung” (American Lung Association, 2016).
To address these concerns, we tested Tetrahymena phagocytosis and ciliary function with
25 mg/mL nicotine e-liquid. Our study used Tetrahymena as a model organism since its ciliary
structure and function are similar to human ciliated cells.
Purpose
This experiment investigates how nicotine-based e-liquid affects Tetrahymena
phagocytosis and ciliary function. We examine Tetrahymena’s ability to swallow ink particles
after e-liquid exposure to determine if it affects respiratory health. The data will help us
understand e-cigarette effects.
Hypothesis
We hypothesize that Tetrahymena’s ciliary motility will be severely impacted by 25
mg/mL nicotine e-liquid, resulting in a reduced rate of ink vacuole intake. We expect
Tetrahymena exposed to e-liquid to have fewer ink vacuoles, indicating decreased ciliary
function.
Methods
Experimental Design
Two groups were tested: one with e-liquid and one with deionized water. The BIOS 312
“Working with Tetrahymena” handout’s modified methodology was used. Both groups have the
following makeup:
Experimental Group (E-Liquid Exposure):
•
10 μL of e-liquid containing 25 mg/mL nicotine
•
990 μL of India ink
•
1000 μL of Tetrahymena
•
Time intervals: 0, 5, 10, 15, 20 minutes
Control Group (Deionized Water):
•
10 μL of deionized water
•
990 μL of India ink
•
1000 μL of Tetrahymena
•
Time intervals: 0, 5, 10, 15, 20 minutes
Data Collection
In both experimental and control groups, we counted ink vacuoles in 20 Tetrahymena
cells at each time interval. We then determined the average ink vacuoles per cell for each group
and time point. This extensive data collection allowed us to examine e-liquid and nicotineinduced phagocytosis and ciliary function.
Results
Figure 1. a. Real Experiment 1
Time
0
5
10
15
20
# of
Average
Vacuoles Cells counted Vacuoles/Cell
20
20 1
Could
not view
20 Could
not view
20 Could
not view
20 Could
not view
20 –
Figure 1. b. Control Experiment 1
Time
0
5
10
15
20
# of
Vacuoles Cells counted Average
260
20 13
273
20 13.65
282
20 14.1
300
20 15
351
20 17.55
Description of Data:
Figure 2.a Real Experiment 2
Time
0
5
10
15
20
# of
Average
Vacuoles Cells counted Vacuoles/Cell
144
20 7.2
116
20 5.8
86
20 4.3
29
20 1.45
0
20 0
Figure 2.b Control Experiemtn 2
Time
0
5
10
15
20
# of
Average
Vacuoles Cells counted Vacuoles/Cell
263
20 13.15
276
20 13.80
280
20 14
307
20 15.35
374
20 18.7
Description of Data:
Table 1 – Experimental Group Results:
In this table, the experimental group was given 25 mg/mL nicotine e-liquid. It delivers
data at 0, 5, 10, 15, and 20 minutes and has two key sets. The first set counts ink vacuoles in 20
cells at each time interval. This data indicates e-liquid-induced Tetrahymena phagocytosis. The
second set shows the average number of ink vacuoles per cell at each time point, giving a more
generalized picture of phagocytosis effectiveness. The number of ink vacuoles changes over
time, indicating Tetrahymena’s ciliary activity in nicotine-containing e-liquid.
Table 2 – Control Group Results:
This table shows the control group’s results with deionized water instead of nicotine eliquid. It also provides statistics from 0, 5, 10, 15, and 20 minutes. The data is divided into two
sets, like Table 1. The first set shows Tetrahymena phagocytosis rates under normal conditions
by counting ink vacuoles in 20 cells at each time point. The second set shows the average
number of ink vacuoles per cell at each time point, revealing phagocytosis efficiency without
nicotine e-liquid.
Discussion
Analysis of results
Our experiment showed that nicotine-containing e-liquid reduced Tetrahymena ink
vacuoles significantly and consistently. It validates our theory and is consistent across time
intervals. The control group, exposed to deionized water, had greater vacuole numbers. This
difference between the experimental and control groups suggests that nicotine in e-liquid can
impair Tetrahymena’s ciliary motility. The decrease in phagocytosis activity suggests e-liquid
exposure may damage ciliary function. This study supports prior findings that nicotine in ecigarettes may cause ciliary dysfunction (Hahn et al., 1992).
Conclusions
According to our study, Tetrahymena ciliary motility and phagocytosis are negatively
impacted by nicotine-containing e-liquid. Reduced ink vacuoles suggests ciliary dysfunction.
More research is needed to confirm these findings and examine how e-liquid components affect
human respiratory cilia (Lerner et al., 2015; Simet et al., 2010).
The results of our study highlight the significance of critically studying the impact of ecigarette liquids on ciliary function, especially when considering their effects on respiratory
health. Our results were consistent across e-liquid components such nicotine, flavouring agents,
and THC, demonstrating the broad breadth of the detrimental effects. Our research illuminates
the mechanics of ciliary dysfunction and e-cigarette dangers. The negative impact on
Tetrahymena urges more research into the processes via which these effects occur.
Explanation of Results
Nicotine’s effects on ciliary beat frequency may explain Tetrahymena phagocytosis’
reduction after exposure to e-liquid with nicotine (Hahn et al., 1992). Nicotine’s action on cilia,
like tobacco smoke, may reduce Tetrahymena’s ink absorption (Cohen et al., 2009).
Diverse Effects of E-cigarette Components
E-cigarette liquid and its components significantly reduce Tetrahymena phagocytosis,
which is crucial to our investigation. Given Tetrahymena’s phagocytosis relies on ciliary
function, this observation is crucial. These results show amazing stability among nicotine,
flavouring agents, and THC e-liquid components. This consistency shows that Tetrahymena
phagocytosis is negatively affected by all elements (Allen et al., 2016; Azzopardi, 2016).
Nicotine’s Role in Ciliary Dysfunction
The negative impact of nicotine on Tetrahymena phagocytosis was a finding from our
investigation that was unexpected. This contradicts nicotine’s ability to increase respiratory
ciliary beat frequency (Hahn et al., 1992). Nicotine exposure reduces Tetrahymena phagocytosis,
raising issues about its processes. E-cigarette liquid composition is complex and may include
complex interactions between components (Chaffee et al., 2018).
The Complexity of E-cigarette Liquid Composition
E-cigarette liquids are complex, despite their appearance. They contain nicotine,
flavourings, THC, and other ingredients. The observed negative effects on Tetrahymena ciliary
function are likely caused by their complex interaction. Understanding these interactions is
crucial to understanding how they impact ciliary function (Fine Maron, 2014).
Potential Mechanisms of Impairment
Future research should examine oxidative stress, inflammatory pathways, cellular
damage, and interactions between e-liquid components in an effort to understand the
mechanisms underlying Tetrahymena’s reduced ciliary function:
1. Oxidative Stress: Oxidative stress markers in Tetrahymena exposed to e-cigarette liquid
can reveal cellular damage and ciliary malfunction (Lerner et al., 2015).
2. Inflammatory Pathways: Understanding whether e-cigarette liquid exposure causes
inflammation and ciliary dysfunction requires studying inflammatory responses (Leopold
et al., 2009).
3. Cellular Damage: To further understand how e-cigarette components affect
Tetrahymena, DNA damage and membrane integrity can be assessed. Navarrette et al.
(2012).
4. Interactions Between Components: The effects of nicotine, flavouring agents, THC,
and other e-liquid components can be better understood by studying their interactions
(Hiemstra & Bals, 2016).
5. Time-Dependent Effects: The temporal effects of e-cigarette liquid exposure on ciliary
function must be studied. Understanding if prolonged exposure changes findings can
illuminate chronic respiratory health issues. (Sisson 2007).
Flaws and Limitations
Our experiment was flawed because cell observations at 5, 10, 15, and 20 minutes were
unavailable. The early use of too much e-liquid caused observation issues. Future tests should
use a reduced e-liquid volume (10 L) and adjusted microscopy settings to address this. These
enhancements will save time and ensure complete data collecting.
Implications
This investigation suggests that e-liquid, especially nicotine-containing liquid, may impair
ciliary motility. These findings raise concerns regarding the respiratory health impacts of ecigarettes due to the structural similarities between Tetrahymena and human respiratory cilia.
This exploratory experiment shows the need for more research to confirm these findings and
understand the mechanisms.
The experiment’s shortcomings must be noted. We measured ciliary function indirectly
using Tetrahymena phagocytosis. Our findings indicate a considerable impact on Tetrahymena
phagocytosis, however the effect on human respiratory cell ciliary function needs more study.
The complexity of the phagocytic pathway and non-ciliary components must be addressed when
interpreting results.
Future Research Directions
Our study lays the groundwork for future research on the complex link between e-cigarette usage
and respiratory health:
1. Mechanistic Studies: Priority should be given to studying how e-cigarette liquid
components affect Tetrahymena ciliary function. This requires studying these
components’ intricate interactions (Simet et al., 2010).
2. Individual Component Analysis: Isolating the effects of various e-liquid components
can assist determine the key contributors to ciliary impairment. Such concentrated
investigations can provide vital insights into the exact compounds responsible for the
reported effects (Cohen et al., 2009).
3. Complex In Vitro Models: The impact of e-cigarette liquids on ciliary function can be
assessed more directly by moving from Tetrahymena to more complicated in vitro
models that mimic human respiratory cilia. These models link in vitro and clinical data
(Panitz et al., 2015).
4. Animal Models: Animal models, especially those with human-like respiratory cilia, can
reveal the physiological and pathological effects of e-cigarette liquid exposure. These
models can provide better human data and inform future study (Alzahrani et al., 2018).
5. Long-term Effects: Investigating the long-term effects of e-cigarette liquid on ciliary
function is crucial. Continuous exposure investigations can reveal e-cigarette-related
chronic respiratory health hazards (Chaffee et al., 2018).
6. Clinical Relevance: Our findings must be linked to clinical and epidemiological data.
Understanding the translational relevance of our research requires determining if
Tetrahymena effects match human health outcomes (Lerner et al., 2015).
Our findings show that e-cigarette liquid and its components negatively impact Tetrahymena
phagocytosis, suggesting ciliary dysfunction. This study sets the stage for future research on the
complex link between e-cigarette usage, respiratory health, and ciliary function. Future research
will inform public health measures to reduce e-cigarette dangers.
References
Allen, J. G., Flanigan, S. S., LeBlanc, M., Vallarino, J., MacNaughton, P., Stewart, J. H., &
Christiani, D. C. (2016). Flavoring chemicals in e-cigarettes: diacetyl, 2, 3-pentanedione,
and acetoin in a sample of 51 products, including fruit-, candy-, and cocktail-flavored ecigarettes. Environmental health perspectives, 124(6), 733-739.
Alzahrani, T., Pena, I., Temesgen, N., & Glantz, S. A. (2018). Association between electronic
cigarette use and myocardial infarction. American journal of preventive medicine, 55(4),
455-461.
Azzopardi, D., Patel, K., Jaunky, T., Santopietro, S., Camacho, O. M., McAughey, J., & Gaça,
M. (2016). Electronic cigarette aerosol induces significantly less cytotoxicity than
tobacco smoke. Toxicology mechanisms and methods, 26(6), 477-491.
Chaffee, B. W., Watkins, S. L., & Glantz, S. A. (2018). Electronic cigarette use and progression
from experimentation to established smoking. Pediatrics, 141(4), e20173594.
Cohen, N. A., Zhang, S., Sharp, D. B., Tamashiro, E., Chen, B., Sorscher, E. J., & Woodworth,
B. A. (2009). Cigarette smoke condensate inhibits transepithelial chloride transport and
ciliary beat frequency. The Laryngoscope, 119(11), 2269-2274.
Fine Maron, D. (2014, May 1). Smoke Screen: Are E-Cigarettes Safe? Even without tobacco, the
poorly regulated devices may pose unique dangers.
Hahn, H. L., Kleinschrot, D., & Hansen, D. (1992). Nicotine increases ciliary beat frequency by
a direct effect on respiratory cilia. The clinical investigator, 70, 244-251.
Hiemstra, P. S., & Bals, R. (2016). Basic science of electronic cigarettes: assessment in cell
culture and in vivo models. Respiratory research, 17(1), 1-5.
Leopold, P. L., O’Mahony, M. J., Lian, X. J., Tilley, A. E., Harvey, B. G., & Crystal, R. G.
(2009). Smoking is associated with shortened airway cilia. PloS one, 4(12), e8157.
Lerner, C. A., Sundar, I. K., Yao, H., Gerloff, J., Ossip, D. J., McIntosh, S., … & Rahman, I.
(2015). Vapors produced by electronic cigarettes and e-juices with flavorings induce
toxicity, oxidative stress, and inflammatory response in lung epithelial cells and in mouse
lung. PloS one, 10(2), e0116732.
Navarrette, C. R., Sisson, J. H., Nance, E., Allen-Gipson, D., Hanes, J., & Wyatt, T. A. (2012).
Particulate matter in cigarette smoke increases ciliary axoneme beating through
mechanical stimulation. Journal of aerosol medicine and pulmonary drug delivery, 25(3),
159-168.
Panitz, D., Swamy, H., & Nehrke, K. (2015). A C. elegans model of electronic cigarette use:
physiological effects of e-liquids in nematodes. BMC Pharmacology and
Toxicology, 16(1), 1-10.
Simet, S. M., Sisson, J. H., Pavlik, J. A., DeVasure, J. M., Boyer, C., Liu, X., … & Wyatt, T. A.
(2010). Long-term cigarette smoke exposure in a mouse model of ciliated epithelial cell
function. American journal of respiratory cell and molecular biology, 43(6), 635-640.
Sisson, J. H. (2007). Alcohol and airways function in health and disease. Alcohol, 41(5), 293307.
BIOS 312, Cell Biology Laboratory
Group Research Project Report Grading Rubric
SECTION
Excellent
Good
Fair
Poor
Introduction
(20 points)
Well‐researched
context for current
study, including
the question to be
addressed.
Hypothesis is
stated clearly in
If‐Then format and
is explained well.
(10 pts)
Well‐researched
context for current
study, including
the question to be
addressed.
Hypothesis is
stated in If‐Then
format but is not
explained well.
(7‐9 pts)
Contextual
information is
vague or not
sufficiently
detailed.
Hypothesis is
stated but is not
explained well or
is not in correct
format. (4‐6 pts)
Contextual
information is
vague or not
sufficiently
detailed, or
statement of
hypothesis is
missing. (2‐3 pts)
(2 pts) 2‐3 pages in length _____
(2 pts) Relevant background information on Tetrahymena _____
(2 pts) Relevant background information on cigarette/nicotine/e‐cigarette use _____
(2 pts) Variables being tested are defined _____
(2 pts) Existing research on the variable being studied is summarized _____
Subtotal:
/20
SECTION
Excellent
Good
Fair
Poor
Methods
(10 points)
Step‐by‐step
description of
procedure that
could be repeated
by another scientist
is included. (5 pts)
Description of
procedure is
included but some
details are vague or
unclear. (4 pts)
General description
of methods is
included but
important details
are missing. (3 pts)
Would be difficult
for another
scientist to repeat
procedures based
on the description
provided. (2 pts)
(1 pt) 1‐2 pages in length _____
(2 pts) Descriptions of procedures are written mostly in third‐person passive voice _____
(2 pts) Explains precisely how data were collected _____
Subtotal:
/10
SECTION
Excellent
Good
Fair
Poor
Results
(25 points)
Data are clearly
summarized in text
and are presented
visually as tables or
graphs that are
easily interpretable
by readers. (10 pts)
Data are clearly
summarized in text
and are presented
visually as tables
or graphs but
there are minor
organizational
errors. (7‐9 pts)
Data are not clearly
summarized in text,
disorganized or not
presented visually
in tables or graphs
that are easily
interpretable by
readers. (4‐6 pts)
Data are
disorganized or
incompletely
recorded and not
presented in a form
that is easily
interpretable by
readers. (2‐3 pts)
BIOS 312 Independent Experiment Report Grading Rubric
(2 pts) Numbers shown in text or tables have appropriate units _____
(2 pts) Tables and figures are labeled with Table/Figure number and title _____
(1 pt) Table/figure numbering is sequential and follows order in which Results are presented _____
(2 pts) Rows/columns (Tables) and axes (Graphs) are clearly labeled _____
(2 pts) Written mostly in third‐person passive voice (statements like ‘we observed’ are okay) _____
(3 pts) Trends and important points found in Discussion are described in Results section _____
(3 pts) No interpretation of data is included in Results section _____
Subtotal:
/25
SECTION
Excellent
Good
Fair
Poor
Discussion
(25 points)
Clearly summarizes
the data and the
rationale for all
conclusions drawn.
Discusses
applications or links
to other studies in
the literature.
Hypothesis is
accepted or
rejected based on
the data. (10 pts)
Clearly summarizes
the data and the
rationale for all
conclusions drawn.
Discussion of
applications or links
to other studies in
the literature is
weak or limited.
Hypothesis is
accepted or
rejected based on
the data. (7‐9 pts)
Does not clearly
summarize the
data and the
rationale for all
conclusions drawn.
Discussion of
applications or links
to other studies in
the literature is
weak or limited.
Hypothesis is
accepted or
rejected based on
the data. (4‐6 pts)
Does not clearly
summarize the
data and the
rationale for
conclusions drawn.
Discussion of
applications or links
to other studies in
the literature is
weak or limited.
Hypothesis is not
accepted or
rejected based on
the data. (2‐3 pts)
(2 pts) 2‐3 pages in length _____
(3 pts) Discussion points are supported by reference to data, observations, and/or literature. _____
(2 pts) Errors, potential biases, or limitations are explained thoroughly _____
(2 pts) A future experiment is proposed that builds on the Conclusions _____
(2 pts) Purpose of the future experiment is explained clearly _____
(2 pts) Variables to be studied in future experiment are described _____
(2 pts) Future experiment is explained briefly without descriptions of specific protocols _____
Subtotal:
/25
Literature Citations (10 points)
(4 pts) At least 5 primary literature sources are referenced
(2 pts) Sources listed in the reference section are cited in text
(2 pts) Sources cited in text are listed in the reference section
(2 pts) Format of in‐text citations follows (Author, Year, Page)
Subtotal:
_____
_____
_____
_____
/10
Participation in Writing Clinics (10 points)
(5 pts) Two copies of appropriate drafts (5 pages in WC1, 8 pages in WC2, all required sections) _____
(5 pts) Appropriate review of two student’s papers in WC1 _____
Subtotal:
/10
Total Score:
/ 100
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