In Medicine

Presentation by Howard and Ryan

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The Heart

• The heart is a muscular cone-shaped organ about the size of a clenched fist of the same person.It is located in the upper body (chest area) between the lungs, and with its pointed end (called the apex) downwards, forwards, and pointing towards the left.The main purpose of the heart is to pump blood around the body.

• The basic structure of the heart (illustrated above) may be described as follows:• The Heart is divided into separate right and left sections by theinterventricular septum, or

"septum" when the context is clearly that of the heart. Each of these (right and left) sections is also divided into upper and lower compartments known as atriaand ventricles, respectively.

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Biosensors• Based on this pathway:

• Analyte: Sensed particulate (cancer protein, molecule, or antigen)

• Sensing element: a means of determining presence of particulate (binding usually)

• Transducer: sensed particulate concentration (or just existance) is converted into a recordable energy (electrical is the ultimate output, every transduction ends there)

• Data processor: Crude analysis of energy to filter out unnecessary elements (filters and sometimes averaging)

... ...

Amplification Application

Usually involves redox, conformational change

of enzymes, etc

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Biosensors: Examples

• Ex’s of biosensors:– Urea biosensors, blood glucose biosensors, etc.

• Essentially detects associated chemicals with metabolism (i.e. H2O2)

– DNA FETs, lectin-based biosensors, etc.• More ‘biological’ of a sensor – detection based on specific

base pair complements

• Ex’s of biological recognition elements:– Enzymes, proteins, antibodies, DNA, cells, organelles,

etc.• Ex’s of transducers:

– Electrochemical, electrical, optical, mass, thermal,etc.

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Biosensors: Cantilevers

• Intro. Antigens/proteins of cancer bind to receptors to change the way conduction occurs

• Multi-tier cantilever with different receptor types gives specificity to cancer types

(i.e. You see so much of one protein, you can apply it to multiple cancers, since a cancer will

have proteins in different proportions)

Weight or steric hindrance 5

Biosensors: Pipette Nanosensor• Tiny device shaped like a pipette

– Essentially a nanoscale needle with a hollow section running through its interior

• Conductimetric / potentiometric– Based on application, may be either; flow of ions on conduction-recording based

• Comprised of a narrow tube, this device normally directs fluid flow– Point detection usually. Tube allows for free flow and increased conductivity.

• Quartz– Stronger, higher melting point, less electrical noise

• Tip etching of dimensions for specific electrochemical effects– Pore geometry is analogous to the way semiconductor channels work, atomic flow action 6

Pipette Sensing Element• Electrode coating

– In fabrication inside is coated with a layer; sometimes a complentary charge substance is put to bind to electrode, Au, ITO, Ti

• Temporary Binding locations– Nanoparticles or other analytes bind/interact to continuously provide current flow (E cell)

or they bind to block the channel, which gives electronic characteristics changes

• Ex: Lone particle, Oglio-tail, Particle+Oglio tail, and bound segment– Each has own characterisic respopnse for the transduction; bound DNA complentry

segment also very specific

• Bindings and blockage– Attraction of analyte, repulsion (and thus stopping analyte), and channel length effects give

current and voltage responses 7

Pipette Transducer

• Transduction based on changes to baseline

• Application if this has to be specific– Detect from background or quantify (pure-ish sample)

• Amplification and analysis– Signal too small to work with

– Sorting out and identifying substance near-raw data

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Why Pipettes?• Taking advantage of small opening

– Nm scale, relatively small amount of particles trapped

• Piercing membranes

– Nondestructive, get in there, get your measurements while still allowing the normal biology to carry out (even if in vitro)

• Fine, precision detection

– Targetting molecules or particulate for

instance at a pore in the nuclear envelope

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Main Focus:Cancer

• Definition: “a malignant tumor of potentially unlimited growth that expands locally by invasion and systemically by metastasis” ("cancer."

Merriam-Webster's Medical Dictionary. Merriam-Webster, Inc. 05 Nov. 2010. <Dictionary.com http://dictionary.reference.com/browse/cancer>.)

• Statistics Canada:

– 163 529 new cases in Canada in 2007

– 62 439 incidences in Ontario in 2008

– most common (out of primary sites) in 2007: prostate cancer

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Cancer

• How it is caused in general:– By “…alterations of intracellular signal

pathways…”*

– By “any aberration of the function of [protein synthesis] genes might lead…to abnormalities of cellular growth, and…to the uncontrolled growth and structural transformations typical of cancer”*

– By certain genes called oncogenes

• ∴ can be anywhere on the body

* “Cell Communication in Health and Disease.” Howard Rasmussen. W. H. Freeman and Company. Scientific American, Inc. 1991.

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Cancer

• What can cause it or increase risk of it:

–Chemical agents, some bacteria and viruses, UV, radiation, tobacco smoke, genetics, age, immune system problems

• Ex.: can be an issue for organ transplants if immune system suppressants used

– Increased risk: being overweight, eating certain types of food, drinking alcohol

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Microfluidic Devices

• Like many similar bio sensors (to be presented) except a continuous fluid flow will be present

• Nanosensor wire essentially has the same sort of antibodies/proteins/etc present

• Bindings produce change in conductivity, or conductance 13

Microfluidic Purification Chips• Used to capture biomarkers specific to cancer

• Blood samples used for testing: suspected cancer and confirmed clean

• The antigen is picked up when the blood

Passes through. Buffer solution runs through

to clean and perform its role in photo cleaving

MPC

Blood

‘Waste’

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MPC Cont’d

• What ensures that only cancer antigens bind?

• MPC binds capture antibodies to surface

• Specific to groups of cancers

• Sample added

• Contains antigen present in cancers

• Buffer run through

• Allow antigens to bind and provide surface for substrate to interact

• Photocleaving

• Detaches complex to be ‘sensed’15

MPC Cont’d

• UV is used to photocleve the antigens– Or specific wavelength that corresponds to the

complex

• Solution run through the biosensor– Charge on floating complex will give, again, specific

I-V characterisics

Sensor

UV

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MPC Cont’d...

• Concentration of this complex (having negative charge) correlates to increased voltage/current (in the Ids Vds plot)

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MPC Cont’d....• Accomplishments:

• 0.33ng/mL detection minimum

– Well within required minuteness, comparable to other methods

• Label-free– Meaning the sensor element can be applied to any

type of detection, its just the MPC that changes

• Reason for label-free

– Physiogic solutions contain irrlevant chemicals that degrade the

minute active surface areas that are used to detect18

Biosensors: Field Effect Transistor in Detection of P53 Gene

• P53 is the gene most commonly found to have mutated when a cancer is detected

• Goal was to detect the presence of mutated p53 vs p53 wild type

• P53 mutated is the transcription factor responsible for 50% of all cancers

This alone is enough to detect a lot of cancers

• p53 wild is to bind to target binding sequence causes more charge to accumulate at that region on the conductor surface, connected to current differences

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FET and Detection System Creation

• Very noticible electrochemical difference when correct binding occurs

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Biosensors: Optical Nanosensor

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Lectin-Based Biosensors

• Detect cells that over-express sialic acid (a glycan), i.e. cancer cells (lung, liver, prostate)

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Lectin-Based Cancer Biosensors

• Glycans are polysaccharides that can be found connected to proteins on the exterior surface of cellular membranes

• Lectins are proteins that bind to specific carbohydrates, and can also be found on the exterior surface of cellular membranes

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Lectin-Based Cancer Biosensors

• How it works:

– 1) Target glycans of (cancer) cells bind to lectins of biosensor

– 2) {Lectin-Au-Th} biconjugates are used as labels because they bind only to cells that have their glycans binded to the lectins

– 3) Using electrochemical detection, the peak current shows how much the target glycans are expressed, and how many cells express these glycans

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Lectin-Based Cancer Biosensors

• Sialic acid is a good indicator of cancer and cancer progresssion,

• But an increase in sialic acid can also be caused by inflammation

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• Carbon nanotube,gold electrodes, &DNA strand

• DNA sequence found conductance changes

DNA FET Biosensors

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Nanotechnology Can Help

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What is Nanotechnology?

• “a branch of technology dealing with the manufacture of objects with dimensions of less than 100 nanometres and the manipulation of individual molecules and atoms” (Collins

English Dictionary - Complete & Unabridged 10th Edition)

• Includes nanomaterials and nanomedicine

• Largely used for the production of products for children, cosmetics, food, packaging, clothes, etc.

• 800 manufacturers idientified products that are publicly available; new ones hitting market at 3-4 a week

• 1015 products or product lines 28

Nanotech Application Examples

• Molecular imaging

• Self-assembling functional membranes with designed properties, such as catalysis, photoactivity, electrical activity, electrochemical and water-purifying activity

• Live-cell imaging of the movements of viruses, proteins, prions and drugs

• Harnessing biological motors, such as muscle and other motile proteins, for mechanical or electrical energy production

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Imaging and Treating Cancer with Nanotech

• Ex.: use dendrimers (branched molecules) to target just the cancer cells by using the fact that they over-express folic acid (vitamin) receptors– Some branches with folic acid, some branches with

cancer drug (could use for melting cells)– Absorbed by cancer cells– Can use for imaging and for treatment– A study found it to be non-toxic, stable, and easily

made to discriminate between cells

• Can use DNA to speed up process of putting dendrimer complexes together

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Nanotechnology: Quantum Dots & Imaging Cancer

• Definition: a quantum dot is “a nano-scale crystalline structure…that absorbs…light and then reemits it a couple of nanoseconds later in a specific color.” (http://www.termiumplus.gc.ca/tpv2alpha/alpha-

fra.html?lang=fra&i=1&index=ent&__index=ent&srchtxt=quantum+dot&comencsrch.x=0&comencsrch.y=0)

• Can use to light up cancer cells and track their movement very early, and can check if all gone

• Lets you light up multiple groups of cells using the same light source (absorbance frequencies are multiples of each other)

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Nanotechnology: Quantum Dots & Imaging Cancer

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Nanotechnology: Nanorobots

• Microscopic machines that autonomously perform actions that are programmed into them

• Usually individuals

Being individual greatly increases risk of failure and general resource costs.. Benefits to delivering over free floating

• Delivered

Packaged in membranes to protect, cut down on energy, prevent dispersement before reaching site where they’re needed

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Nanotechnology: Nanorobots Cont’d• Power

• How will robots get their power?Can have batteries but those are hard to miniaturize

• An easy way to generate power

is to simply use what resources

are already available

(For example: oxygen be taken

from blood, along with glucose

and produce energy for what the robots need)

• Making use of thermal energy available from nearby reactions

• Electrochemical gradients (cells and ions floating in bod)

• Incorporated batteries not a good idea for the time being

• An example of how this all can be carried out is to have the robots sit on the side of a vessel wall, do their sensing, chemical production while grabbing what they need from passing blood

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• Nanoshell is a nanoparticle made of a thin outer metallic shell

• Core is diaeletric

• Quasiparticles called plasmons are involved which are a collective excitation or quantum plasma oscillation where the electrons simultaneously oscillate with respect to all the ions

• Collective excitation is caused by exposure to specific wavelenth of liight (IR usually)

• What this means is that the cells where the nanoshells are will be destroyed (thermal energy and membrane cracking

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Nanotechnology: Using DNA to MakeCarbon Nanotube FETs

DNA “scaffolds” for self-assembly

DNA nanotechnology

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Issues in Nanotech

• Buckyballs and carbon nanotubes

• Nanotech usually just miniturizing existing engineering and putting those things out in mass amounts

• No accounting for the large scale effects... Connotations such as ‘grey goo’ arise

• Possibility of unavoidable surveillance by tiny devices

• Weapons

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Issues in Nanotech cont’d

• Government regulations on size, mass, toxicity, etc.

• Possible lack of understanding in the general public

– need research to evaluate risks

– nanotech has already been around for years, ex.: silica & clay nanoparticles in food and food packaging

• Sometimes onflicting definitions of nanotechnology – complicates policy-making

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Future: Nanotechnology and Biosensors

• Efficient: smaller, cheaper, faster

• Targeting

• More extensive use of self-assembly

• Self-powered: nanogenerators

– Vibrations, temperature gradients, biochemistry, ultrasonic waves, noise, hydraulic energy, etc.

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Future Alternative:Dog Biosensors

• Detect biochemical markers in urine/breath

• Can be pretty 88~97% accurate – some mixed results (dog type)

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Future: Goals and Focus of Nanorobots

•Biomimicry•Ex. Based off bacterial flagellum•Membrane protection

•Push towards miniutrization•Power issues, some circuit components can’t be scaled down•Immune responses

The key aspects of what lie ahead in nanorobots consist mainly, theoretically, in biomimicry. What we want to do is take a human approach to what nature already has produced. So an example would be this motor here, which is inspirationally derived from the bacterial flagellum. The capacitor here can be charged by internal body potentials or ion flow, or its own power source from fuel it collects as it passes through.Just to explain the rest of this diagram here: you have a camera here to direct the robot, or detect the payload location, and the payload is the chemical, for instance to destroy the cancer.What we essentially want to do, since its easiest to conceptually design, is miniturize existing technology, such as circuitry, robotics, and processing systems (such as how the robot will decide to move).

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Too large! Clogs body systems!

Future: Limitations

The limitations in all this consist of the fact we're appyingconcepts we're already familiar with in our big macroscopic world, hoping it will work on the microscopic world. However, there are issues concerning the damage these machines will have on the body: depection of nutrients, colliding with vessels, disrupting the body's natural processes, and even causing blockage. This picture here shows the naive view of how it will be... a robot , for instance the size of a cell trying to do away with it, but in reality, this is a big bulky stupid idea. We want lots of little things that won't interfere with the body at large.

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The End!

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