The only students who ever tried to defend me against harassment at Bunker Hill Community College were a black male and a black female. That was in my Chinese class, last year.
The only reason that it was a Chinese-American student who instigated the harassment in that class was that he already spoke Mandarin Chinese. He already knew everything that the professor was trying to teach to other people, and he didn't care that he was disrupting the class for everyone else who didn't already speak Chinese. It wasn't about race; it was about gender, and the conglomerate having ruined my life by targeting me for bullying. I'm sure that minority girls and women are being targeted for harassment as much, if not more, than white girls and women everywhere.
The reason that the Chinese, female professor for the class failed to stop the harassment and didn't know what to do was that the school had failed to educate her about the issue.
This is a homework assignment from the Biology class that I had this year, before the summer:
_________________________________________________________________________________
Name: Lena Kochman
Article Assignment 2. Remember to cite your sources INLINE.
Part 1: Biology definitions. Fully understand what each of these terms
mean. Most of these concepts can be found in your textbook and shouldn’t take
too much time.
1. Endogenous gene: An endogenous gene is produced by the
organism in which it is found. (https://en.oxforddictionaries.com/definition/endogenous)
2. Exogenous gene: An exogenous gene is introduced into an
organism from a foreign agent.
3. Locus/loci:
Another word for “location”; in this context, it describes where a
gene or a mutation is on a chromosome.
“Loci” is the plural form. (https://en.oxforddictionaries.com/definition/locus)
4. Nuclease:
An enzyme that breaks bonds of nucleotide chains. (https://en.oxforddictionaries.com/definition/endogenous)
5. Ribonuclease: An enzyme that breaks apart RNA. (https://en.oxforddictionaries.com/definition/rnase)
6. Heterologous gene expression: The expression of genes that were
introduced into a cell and that are not native to the cell.
7. Non-homologous end joining: A method by which the cell repairs broken
DNA. Also called “NHEJ.” (https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3758668)
8.
Cell
transfection: When specifically
genetic material, such as DNA or RNA, are introduced into mammalian cells.
(https://www.jove.com/science-education/5068/an-introduction-to-transfection)
9. Double stranded break: When both strands of DNA are broken at
the same place.
10.
Nicked
DNA: A break in one strand of DNA
(https://www.neb.com/applications/dna-modification/dna-nicking)
11.
in-vitro: A process that happens in a test tube or
otherwise not in a living organism (https://en.oxforddictionaries.com/definition/in_vitro)
12. in-vivo:
A process that happens in a living organism
Part 2: Experimental techniques and reagents. Explain WHAT these are
and WHY they are used in research.
1.
DAPI
nuclear stain: A fluorescent stain
used to locate DNA in cells. (http://medical-dictionary.thefreedictionary.com/DAPI+stain)
2.
Green
fluorescent protein and mCherry: These
are marker proteins that are produced when the nucleotide sequences that code
for them are introduced into host cells.
One is green and one is red. (https://www.thermofisher.com/us/en/life-science/cell-analysis/fluorophores/green-fluorescent-protein.html), (www.clontech.com/US/Products/Fluorescent_Proteins_and_Reporters/Fluorescent_Proteins_by_Name/mCherry_Fluorescent_Protein)
3. 293FT cells: These are one of several types of
mammalian cells that are commercially produced for experimentation. They are cloned from human, embryonic kidney
cells. (https://www.thermofisher.com/order/catalog/product/R70007)
4. Expression vector: A carrier used for the expression of
cloned genes. (http://groups.molbiosci.northwestern.edu/holmgren/Glossary/Definitions/Def-E/expression_vector.html)
5. DNA construct: A sequence of DNA that has been created
or modified in the lab for introduction into and transcription by a cell. (https://definedterm.com/dna_construct)
6.
Sanger
sequencing: A method of determining
the sequence of the bases in DNA. (https://definedterm.com/dna_construct)
Why these techniques and reagents are used in research:
If you’re working with DNA, you
need to know where it is in the cell. If
you want to know how a gene is expressed, using a reporter gene that results in
some or all of the cell or organism being turned red or green can help
you. To experiment with biological
processes, you need to have biological substances to study, so you need a
standardized supply of cells. A vector
occurs in nature and its gene-delivery process is recreated or imitated in the
lab to observe how genes work or to carry out an experiment of which the goal
is to change a cell’s genome and gene expression. There are scientists who study subjects
because they think they’re interesting, but many scientists also study things
because they want to be able to control the processes that they’re studying. Some people study bacteria because they think
bacteria are fascinating; others because they want to develop antibiotics. A DNA construct was an inevitable result of
genetic research, as was a method of identifying naturally occurring DNA sequences.
Part 3: Understanding the paper.
1. What does “genome editing” mean?
Genome editing is a process that can take a number of
forms. Essentially, it’s the removal
and/or addition of base-pair sequences from/to DNA to create or remove a
mutation.
2. What are the potential benefits and
downsides of genome editing?
A potential benefit of genome editing is the curing of
diseases that have genetic causes. (http://www.yourgenome.org/facts/what-is-genome-editing)
The potential downsides are pretty much everything else. Something can go wrong and cause a serious
mutation that kills someone or that is passed to that person’s children and
causes problems for future generations. There
is also the question of life occurrences that some people think are detrimental
and some don’t. Is it bad to get
old? Is it bad that everyone has to die
eventually? For diseases that have
subjective diagnoses, what are the ethical boundaries for treating people who
are said to have them? What about
genetically engineering fetuses at the request of their parents? What about genetically engineering children
who are already born, at the request of their parents? What about people who will want genome
editing to serve the frequently questionable function of plastic surgery? Do they deserve to have elbows growing out of
their ears? It’s all really a scientific
and moral conundrum.
Introduction:
3. What does the acronym CRISPR stand for?
“Clustered regularly interspaced short palindromic repeats.” (www.yourgenome.org/facts/what-is-genome-editing)
4. In order to cut DNA both in-vitro and in
prokaryotic cells- what are the 4 genes you need to express?
“TracrRNA,
pre-crRNA, host factor ribonuclease (RNase) III, and Cas9 nuclease.” (Cong, 2 (820))
5. Why is it important to add an NLS when
doing these experiments in mammalian cells? (Why wasn’t it necessary in-vitro
or in prokaryotes?)
You need a nuclear localization signal, or NLS, for
experiments with mammalian cells because the genetic material in a mammalian
cell is in the nucleus and separated from the rest of the cytoplasm by the
nuclear envelope. In-vitro, the only
things that you’re working with are whatever you added to the test tube or
other container, and in a prokaryotic cell, the genetic material is accessible,
being in the cytosol with everything else that’s in the cell. (http://www.jbc.org/content/282/8/5101/.long)
FIGURE 1:
Panel A. Testing DNA constructs:
1. What does NLS stand for and what is its
function?
Nuclear localization signal.
It guides the CRISPR into the cell’s nucleus. (http://www.jbc.org/content/282/8/5101/.long)
2. Which SpCas9 construct worked the best?
It looks as if locusSpRNase III mCherryNLS worked the best.
3. What is the evidence that shows it?
There is the most color expressed from that construct.
Panel B. Experimental setup:
1. Which of the 4 pictures represents the
pre-crRNA?
The one that has this sequence: U6 DR EMX1(1) DR
Panel C. Experimental setup:
1. What does PAM stand for?
Protospacer adjacent motif.
(https://genomebiology.biomedcentral.com/articles/10.1186/s13059-015-0818-7)
2. What is the PAM nucleotide sequence used in
this paper?
GGG
Panel D. Testing the system:
1. Look up surveyor nuclease assay on
Wikipedia.
a. Smaller bands are [higher/lower] on the
gel?
Lower.
b. Smaller bands indicate [digestion/no
digestion] of the DNA?
Digestion.
c. For this paper, digested DNA indicates
[success/failure] of experiment? Why?
Success. The
experiment was about the success of CRISPRS at breaking DNA at targeted
sites.
2. What is the positive control in this
experiment? Explain.
SpCas9. It’s present
in every well.
3. What does this experiment tell us about the
need for SpRNaseIII? What is the data that shows this?
“SpRNase III was not necessary for cleavage of the
protospacer (Fig. 1D).” (Cong, 2
(820)). In the absence of SpRNase III,
DNA cleavage occurs, as shown by the separated bands in the 4th well
in Figure D. The 4th and 5th
wells are almost identical.
Panel E. Sequencing results:
1. What does indel mean?
“Small insertions and deletions.” (https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2953750)
2. Did this experiment show insertions,
deletions, or both? Explain.
Both. The goal of the experiment was to demonstrate
the versatility of a CRISPR system to create change at several loci at one
time. The diagram shows that there were
both insertions and deletions.
This is a quote
from the article which seems to indicate that the experiment was successful
both at creating deletions and insertions:
However, nicked DNA can in rare cases be processed via a DSB
intermediate and result in a NHEJ event (27). We then tested Cas9mediated HDR
at the same EMX1 locus with a homology repair template to introduce a pair of
restriction sites near the protospacer (Fig. 4C). SpCas9 and SpCas9n catalyzed
integration of the repair template into EMX1 locus at similar levels (Fig. 4D),
which we further verified via Sanger sequencing (Fig. 4E). These results
demonstrate the utility of CRISPR for facilitating targeted genomic insertions. (Cong, 4 (822))
_________________________________________________________________________________
August 2, 2017
The Biology class was taught by a female, minority professor. It was the most academically difficult class that any of the students had taken at the college. I don't know if she'll be teaching there again this year; the conglomerate's penchant for hacking my school email and website accounts and attacking everyone who doesn't abuse me might have prompted her to take employment elsewhere.
The Precalculus class in which I was harassed by three male students, before the summer, was taught by a white female. She'd taught there for 11 years and never had an evaluation; the school has no idea who's a capable teacher and who isn't, because it doesn't evaluate anyone other than asking students to do multiple choice surveys at the end of each semester. She was one of the best math teachers that I've ever had. She had already decided to leave Bunker Hill Community College before the semester started; she'd taught there while also being a full-time teacher at a high school. The Spring 2017 semester was to be her last semester before being a mother. The three male students who harassed me in that class were not the best students; they were students who didn't care about learning the subject, and they didn't care about disrupting the classroom. They weren't the only students in the class who weren't white; most of the students in that class weren't white, and most of them didn't harass me. It doesn't take more than one loud, persistent cougher in a classroom to be disruptive, particularly when the person doesn't stop no matter how much he or she is ignored. This is a harassment by people who want to provoke a reaction; they think it's funny, and the conglomerate has told the world to think it's funny for 7 years. They think it is the funniest thing to provoke someone until she says "Please stop" and then to act as if they're not doing anything and their target is crazy. They don't stop when they're ignored, and they don't stop when they're asked to stop by their target; they stop when they're told to stop by someone whom they have to respect.
The French class from which I was forced to withdraw by the school's administration this summer was taught by a black female. SHE WAS FROM PARIS; THE REAL PARIS! I'D NEVER HAD A FRENCH PROFESSOR WHO WAS FROM FRANCE BEFORE! NOBODY CAN TELL ME THAT THE CONGLOMERATE ISN'T DESTROYING EQUAL OPPORTUNITY FOR ME! Obviously, nobody had told her that it wasn't my fault that the Chinese professor had left the school the previous year, which is probably why she was nervous about having me in class. It could also be that her mood about working at the school wasn't optimal because she would have had a higher salary working for minimum wage at employment that doesn't even require a high school degree. She has an advanced degree and has worked as a translator; she speaks French, English and German.
Ultimately, it wasn't the French professor who was the final catalyst for my being suspended from school. It was the school advisor whose inaccurate depiction of me to the Dean's office facilitated the Dean of Students and the Director of Student Services exploiting the opportunity of my reporting being harassed for the third semester in a row of attending the school to say that I was lying and that I was the one who was trying to intimidate other people. They did this to try to excuse their negligence and inappropriate punitiveness toward me from last year, which was the first time that I was falsely counteraccused for reporting being harassed. All I had asked was that the student who harassed me in the Spring 2016 semester be told to stop; it took the school 6 months to do that, and the school only finally did it because the school's failure to do it in April 2016 when I first reported the harassment emboldened him to stalk and harass me online in September 2016. When I provided the irrefutable proof that he had sought me online to harass me and that he had used his first and last name, there was no doubt who he was and that his behavior toward me was unprovoked; that's when the school said that his behavior would be stopped. There was no apology to me from the school. There's also been no apology to me from the conglomerate, which treated him like a hero and whose praise of him encouraged him to stalk and harass me online in September 2016.
The school advisor is a professor of psychology and a minority female. It could be that her mood also isn't optimal most of the time when she's at school; many of the people who work there have to work in tiny cubicles that have no windows and no privacy from anyone around them. When I met her last year, and until the summer of this year, she was working in an office that only had a partial ceiling. Although she also counsels students, everything that she or a student said in her office could be heard by people in the hallway and the person on the other side of the partial wall between her office and the office of the person in the next cubicle. It was also my impression that she was targeted by at least one corporation because she had provided support to me over the past year. She probably didn't want her life and career ruined by the powers that persecute me.
This is a link to the demographics page at the Bunker Hill Community College website:
http://www.bhcc.mass.edu/about/institutionaleffectiveness/fastfacts/
These are pictures of consecutive sections of that page:
The page says that 56% of the students are women, 43% are male, and 61% are students of color.
All you have to do is spend a few days in a classroom at that college to know that the public school system that is supposed to educate students from kindergarten to the end of 12th grade is a national disgrace. If there are more women and students of color than white men at the school, that is because they are denied access to other colleges in higher numbers than white men, even white men who are the products of that same disgraceful public school system.
Unfortunately, access to a privileged education isn't a guarantee that someone's subsequent contributions to the world will be positive.
These are pictures of sections of the first page of the New York Times' website, both consective and nonconsecutive:
That's the New York Times hacking my phone, email and laptop. I had to spend the rest of my financial aid for the Spring 2017 semester on a laptop from the school's bookstore. All of my classes required a computer and I couldn't be at the school's computer labs without being harassed.
That's the New York Times hacking my phone, email and laptop. I had to spend the rest of my financial aid for the Spring 2017 semester on a laptop from the school's bookstore. All of my classes required a computer and I couldn't be at the school's computer labs without being harassed.
___________________________________
That's the New York Times excusing all of the illiterate celebrities who have spent 7 years promoting crime and making my life hell.
___________________________________
That's the New York Times attacking two of my female professors at Bunker Hill Community College, and promoting voyeurism, involuntary pornography and child molestation.
The New York Times probably hacked my phone and my laptop this morning and watched me write all of this page before I started to read the first page of the New York Times' website.
___________________________________
That's the New York Times calling me stupid for being homeless, even though the New York Times is one of the many media sources that told the world to target me for voyeurism, that tapped into the hidden, illegal cameras in both of my last two apartments and that watched and heard live, illegal video of me in those apartments, and that ridiculed me every day from January 2017 to May 2017, while I begged that someone who knew that the voyeurism wasn't a false accusation corroborate that it wasn't a false accusation so that I would stop being treated as if I were a crazy liar by the property manager, the property management's lawyers, the Cambridge police who did nothing to investigate, the court system, the Cambridge Inclusionary Housing Program, Cambridge and Somerville Legal Services, Boston Medical Center, On The Rise, The Pine Street Inn, HomeStart, and every other agency that I approached for help, including the Boston Area Rape Crisis Center, where the white, twenty-something female advocate who was not a lawyer and who hadn't read most of the information that I had given to that office the day before the meeting did more harm than good when she called the property manager and the property management's lawyer to try to prevent my having to be homeless for the third time since 2010.
There is not one person whom I approached for help in the Boston area about the voyeurism in my last apartment who wouldn't have helped me if he or she had had corroboration that I was telling the truth. While they all did what they could to try to help me without that corroboration, the conglomerate amused itself by watching and hearing me not only from the hidden, illegal cameras in my apartment, but also at my phone and email where I called and wrote and read messages to every nonprofit agency for months. I spent the entire Spring 2017 semester futilely fighting an illegal eviction in which I was treated as if I were a dangerous, psychotic criminal instead of the victim of a crime, while the conglomerate laughed.
I have never cared about the conglomerate's opinion of me; it is a worthless opinion. Unfortunately, the worthless opinion of powerful people can ruin lives.
That's another of the vagina jokes that I have told the conglomerate to stop making since 2010.
___________________________________
That's the New York Times promoting voyeurism, involuntary adult pornography, child pornography, and raping children in parks.
The New York Times is also attacking and ridiculing my old friends, which is what the conglomerate began doing when perpetrating the same rumors about me every day began to be dull, even for the conglomerate.
I don't choose excerpts or supporting information for code purposes.
Copyright, with noted exceptions, L. Kochman, August 2, 2017 @ 10:24 a.m.
