training program 2012-2013 - Istituto Pasteur

importantpsittacosisBiotechnology

Feb 20, 2013 (4 years and 6 months ago)

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ISTITUTO PASTEUR
-
FONDAZIONE CENCI BOLOGNETTI

Science at school

PROJECT 2012
-
2013



Program for students

Each l
aboratory experience

normally
lasts
one day, with the possibility to
return

after 15 or 30 days

if a further

observation is required. The experiences are focused on topics within the following
educational research lines:

1)

Microbiology and immunology

One hour lesson about the history of pasteurian sciences and their importance


Experience 1: Spontaneous
generation

Objective: to demonstrate that living organisms are generated only by other living organisms

Experience:
disprove the theory of
spontaneous generation

through

pasteurization


Experience 2: Bacteria world


An introduction

M
icrobial
c
ulture

techniques.

Microscope observation of bacteria and
analysis of
b
acterial
resistance to a
ntibiotics




2) T
he cell cycle


One hou
r lesson
on

how cells divide:

mitosis and meiosis. C
h
romosomes preparation and
observation.

Experience 1: Mitosis

Objective:
to
allow students to pr
actice some of the up
-
to
-
date t
ec
h
niques used in cell biology
such
as

in vivo

fluorescent
imaging
.


Experience: mounting of microscope slides through air drying
technique

using

drosophila

cells
that express tubulin and histon
e
proteins

conjugated

with fluorescent

markers

(Green Fluorescent
Protein,

Red Fluorescent Protein). The
samples

will

then

be observed
under

the microscope to
show

dynamic behavior of the mitotic spindle

and chromosomes during cell division.




Experience 2:

Chromosomes

Objective:
To explain

and

show students

chromosome


morphology

in

the
different

phases

of the
cell cycle
.

Experience:

preparation and


observation

under

the

microscope
of

mitotic,
interphase

and

meiotic

Drosophila

chromosomes
.

Experience 3: B
lood and bone marrow

Objective: Explain
ing

blood and hematopoiesis
,
formation of blood cells (red cells, white cells and
platelets) from the bone marrow. In
troduction to progenitor cells (
stem
cells) and to their site
s

of
origin

(
i.e.
,

the umbilical cord). Introductio
n to blood groups.

Experience: m
icroscopic observation of blood
cells
and bone marrow
.


3)

G
enetics and heredity

One hour lesson about
Mendel's l
aws, the concept of gene and

the

mechanisms of
heredity
.

Experience1: M
utation

Objective: to show that genetic variability in population can


resu
lt
from


DNA

replication
errors
(
spontaneous or induc
ed
)
.

Experience: observation of the different
development
stages and

phenotypic characteristics of
the
fruit flyes
,
observation and analysis of several different mutants.


Experience 2: Mendel’s l
aws and sex
-
linked inheritance.

Objective: to
explain

Mendel’s Laws

through the u
se of drosophila instead of pea

plants.

Experience: Cross homozigous flies exhibiting a phenotypic mut
ation, wait for two generations,
observe

and select

the progeny
with different phenotype and analize the results.


Experience 3:

R
ecombination

Objective: To

show students

why


children

are
never identical

to their parents.

Experience:

cross

flies

aristaless

(without

antennas)
,
dumpy

(
short side) and
black

(
black body)
with

wild
-
type

flies
, wait

for

two
generations
,

identify

offspring

with a different

phenotype and

analyze the results.


Experience 4
:
Genetic

variability

Objective: To

show students

an example

of genetic variation

between individuals

referring to the

genetics of taste

in humans.

Experience
: To provide
students

with

some

harmless substances
, such as sodium
benzoate
, aloin,
s
alicin

or

prop,

the taste of which

varies

depending on
one’s

genetic make
-
up
,
make a


population

assay

and analyze the results
.




4) B
iotechnologies

One h
our lecture

on the discovery of

DNA
,
molecular biology


techniques

a
nd
their

applications

in
basic

and applied research
.


Experience 1:

DNA

extraction

from mouth

mucosa cells.

Objective: To

show students

the

filamentous

nature

of the

nucleic acid molecules
.

Experience:

DNA extraction

from cells

contained

in the saliva

or

hair follicles.

Each
student is

involved in the collection

of

saliva and

in

all the steps

that lead

to the extraction

of DNA

from the
cells

contained therein
.

The

DNA

thus produced

can be

taken away and

stored for

a long time

or
further analyzed
.

Experience 2:

The

PCR

Objective:

t
o show students

how to perform

the technique that

has revolutionized

molecular biology

and biotechnology
:
the
PCR

(Polymerase

Chain Reaction)
.

Experience:

Each student will be

involved in the collection

of
her/his own

saliva sample
, extraction
of the


DNA

from the

mouth mucosa

cells


and

its

"amplification
" using the
PCR reaction.

A

particular stretch

of chromosome 16
,


which is

very

"variable"

in the human population
,

will be
amplified.

In

this way
students

will

become familiar with some

fundamen
tal

aspects

of genetics

(
i.e.
,

homozygosity

and

heterozygosity
)
and

population genetics

(
i.e.
,

the gene frequency
).
Furthermore

they will be
taught to perform

one of the

most
used

molecular biology techniques
:
the
DNA fragments

separation

by

gel
electrophoresis
.

Experience 3:

The

DNA

"fingerprint"

Objective: To

show students how to

get maps

of

DNA

that can identify

a single among a
population
, as does
a fingerprint

at a macroscopic level

(
hence the term
"fingerprint")
.

Experience:

DNA samples

(one

hypothetically

taken from the

crime scene

and five

belonging to

potential suspects
)
are digested

with

restriction enzymes
,
that

recognize and cut
specific

DNA
sequences

that

differ


the

individuals

of

a population
.
The

"view" of

DNA

and its diversity

among



people

will be

obtained

through one of the

most used techniques

in molecular biology
:
the

DNA

gel
electrophoresis.

In

this way, students

will be able to

directly

identify the potential

"murder
er
"

among

the five

"
suspects
"
.

Experience 4
: Production of
transgenic organisms

(1)

Objective: To

show students

how to produce

a

Genetically Modified

Organism

(GMO).

Discuss
how the

use of

genetically modified

animal models

has

allowed us to understand

the function of
genes

(
during

embryonic develo
pment and

in the maintenance

of tissue homeostasis
).

Experience:

observation

of genetically modified mice
;
visit

to the
animal department

and

associated
structures
;
observation of

cell cultures

obtained from

wild
-
type

and transgenic

mice
.

Experience

5:
Production

of transgenic organisms

(2)

Objective: To

show students

how to obtain

genetically modified plants

which
are able to

resist
disease.

Experience:

Students will

wittness

some of the

main steps

of the procedure

used to generate

a

genetically
modified plant.

E
xperiments

will

demonstrate

that


transgenically modified plants


can

activate

better

defense responses

and are

more resistant to infection
,

while keeping their

original
features.


5) B
iology of evolution

One h
our lecture

on the
structure,

function and evolution

of living organisms.

Introduction to

model
organisms.


Experience 1:

Mutation

Objective: To

show students that

the variability

of individuals within

a population,

and between

populations
, may be due
to errors

in DNA,

both
spontaneous and

induced.

Experience:

observation

of developmental stages

of flies
, identification
of mutants

at

the

stereoscope.

Experience 2:

Model organisms

Objective: To

show
students that

despite
striking

morphological differences
,
different organisms

share

genetic and

molecular

features
evolved

from a common ancestor
.

Experience:

observation of some

model organisms
:
the

Enterobacter

Escherichia coli
,

the yeast

Saccharomyces

cerevisiae
,

the nematode worm

Caenorabditis

elegans
, the plant
Arabidopsis

thal
iana,

the fruit fly

Drosophila

melanogaster

and

the mouse.
Introduction

to bioinformatics.

Experience 3:

Living Organisms

Objective: To

illustrate

mechanisms of evolution

focusing

on adaptations

related to

locomotion.

Experience:

visit the

Animal Park

of
Piano

dell'Abatino

(
www.parcoabatino.org)
to observe the
animals

in relation to their

gait

(
biped and

quadruped)
,

brachiation

and other specificities

(i.e.
,

kangaroo
, bat,
emu and

more).


It will be explained how

natural selection

has favo
u
red

different

forms of

ambulation
, referring to the
phylogeny
.

Experience 4
: Evolution

of

mechanisms of

regeneration

and repair.

Stem cells
from

the myth

of Prometheus

to regenerative medicine.

Objective: To

discuss the

cellular and molecular mechanisms

that underline

regeneration

of

damaged tissue

and repair
, trying to
answer these questions
: What
distinguishes a

tissue

able to
regenerate

from another

with regenerative limitations
?
Why do some

species (
Hydra,
amphibians,

urodeles
)
regenerate

better than others
?

Stem
cells

biology,
therapeutic potential

and


limitations.

Discussion about how the

use of

genetically modified

animal models

has

allowed us to understand gene

functions

(
during

embryonic
development and maintenance

of tissue homeostasis
).

Experience:

observation

of genetically modified mice
;
visit

to the animal
department

and

associated structures
;
observation of

cell cultures

obtained from


wild
-
type

and transgenic

mice
.

6) Neurophysiology

One hour lesson on

the

mechanisms

of communication between

cells

of the nervous system
.


Experience 1:

Use of

frog

oocytes

of

Xenopus

Laevis

to study the function

of

receptors

for

neurotransmitters
.

Students can

assist to the isolation of

frog

oocytes
,
the

microinjection

of

cerebral membranes
, the
isolation
of
membranes,

the recording of

ionic currents

induced

by stimulation

of

GABA receptors


"

microinjected
"
in oocytes.


Experience 2:


Measuring

the

concentration of

intracellular calcium in

neural in vitro culture

methods
.

It will be

possible to observe

neurons

derived

from the hippocampus

of mice

that have been
coloured with

immunohistochemical

methods

in order

to recognize

different cell
types

(
neurons,
astrocytes,

microglia)
.
Students will attend

experiments

stimulating

metabotropic

and

ionotropic

n
eurotransmitters receptors

and
observe kinetics

differences

in the

in
crease

of

intracellular calcium
concentration

depending on whether

this

comes

from outside or inside

the cell
.





Experience 3:

Fluorescent

observation of transgenic mice

brain cells movement.

Brain tissues

from

transgenic animals

expressing the

fluorescent protein

GFP (green

fluorescent
protein)

in


specific cell types

will be used
to lead

experiments

of
microstimulation

with

ATP

(
adenosine triphosphate)
and

observe the
movement

of

microglial cells
.






7)

Ecology

Objectives: to illustrate the fate of a dead leaf. Which organisms are active in the process of
decomposition? (each student can conduct an experiment with microbes and
animals
)

Experience 1: Decomposition

-

Observation in capsules of what happens to leaves that have just fallen off a branch.
Focus

on (i)
the process of demolition of organic matter in its two principal phases (ii) the biological operators
of the process o
f decomposition (micro fungi and invertebrate detritiv
ores).

-

Simplified calculation of the decomposition rate for biological materials of different origin
(vegetable and animal). The student will be able to calculate the time necessary
for

the

complete

biological demolition of complex

substances (for example lignin
)
.

Experience 2: Ecological balance

Preparation of micro
cosme

with primary producers (micro algae) and consumers, for researching
numerical relations between species that maintain a balance b
etween production and consumption
in their ecosystem.








8
)

F
ood and

nutrition

One hour lecture

on the origin of

food science

and human nutrition
:


Food and

natural history of humanity
.

Food that

originates

calories and

non
-
caloric

foods

(
dietary fiber)
.

Foo
d

obtained by

fermentation
.

Health

food
.

Functional food
.

Intolerances

and food allergies
.

Foodborne

diseases
.



Experience 1:

F
ood and the processes of aging

Objective: t
o

show students

that

food is

in fact

a tissue

of biological origin and

for this reason is

subjected to

metabolic processes

of aging,

some of which

are in
conflict with

their function

/

nutritional value.

Experience:

half a day
experimental activity



on


Maillard reaction
,

AGE

products

and

other
enzymatic reactions

involved during the

food
transformation
process
es.


Experience

2.

Food design and preparation

Objective: To

explain how nowadays


knowledge of the principles

of eating and nutrition

is

lead
ing

towards the d
esign of

novel food.

Experience:

Production of

a soft drink

(i.e.
,

coca
-
cola

similar)
.


In addition to the

training courses

the following seminars


will be organize
d
.


These will be

especially

dedicated to the
students

attending

the last two

years of

high school.


Depending on the number

of the participants
,
the

conference

can

also take place

at school.

------------------------------------------------------


SEMINARS

Immunology

-

The immune response

and

immunological

based diseases



Microbiology

-



How

bacteria

can

take the wrong way

The acquisition of virulence genes

by

horizontal gene transfer

via

plasmids, transposons
,
integrons
and

PAI

to trace the

evolution

of pathogens

from

nonpathogenic

ancestors
.



Bacteria in society: the biofilm
s


How a microbial community grows, how
bacteria
communicate and what the advantages of living
in biofilms

are
.




The terrible

story of Dr.

Semmelweis


Paradigm

of an innovative scientific discovery



During the conference the following topics will be
developed
:



The basic method

of
a
discovery,

description

and explanation

of the causes

of the phenomena
.



The Phenomenology

and molecular mechanisms
.



The impact of technology
.



Microbes

before

Pasteur.



Experimental models
.

• Communication

of a disco
very.

• Discovery

and common sense.

• Why

the "
Academy" sometimes
reacts badly
.

• Tools

and

methods of communication.



How to

react

to a "
new
culture."

• Trust

and

distrust
: Error,
Fraud and / or

Great discovery
.

Teachers

program

• Seven day

courses with

formal

lectures

on

Genetics
, Genetics and
Developmental Biology
,
Human Genetics
, Molecular biology
and

evolution,

Ecology and

Neurobiology,

accompanied by

simple

experimental procedures

to be proposed

in the classroom.




One day courses on topics

of
social interest
:
genetically modified organisms
, stem cells,
cloning,

genetic diseases
, cancer,
neurobiological

effects

of drugs and alcohol
, nutrition and
metabolism,
etc.
.



Formal genetics
:


Mendel's laws

and heredity
.

The

concept of
gene

and

the

chromosomal theory
.

Model organisms

to study


heredity mechanisms
.

Mutations
, recombination
and

genetic mechanisms

in the evolution

of genomes
.

Developmental genetics
:


Basic concepts

of development
.

Model organisms
for

development
al studies
.

Construction
of the

body
pattern
.


Evolutionary developmental

biology
.


Human genetics:


Genetic variability

in humans
.

Identification

of a

genetic polymorphism
.

Evolutionary mechanisms

in humans
.

Molecular biology:


DNA:
everybody talks about it
, but what is it like
?

DNA structure and

its

functional implications
.

Structure and

function of

genes
.

From gene to protein
.

DNA manipulation
.


Biotechnology:


Biotechnology:

diagnostic forensic and

paleobiological applications.


The

PCR

(Polymerase

Chain Reaction
)
,

a
Nobel
Prize technique
.

Polymorphism of

Alu sequences

and

DNA fingerprinting
.




G
enomics:


Structure and complexity

of the genome
.

Genome sequencing
.

A function for

each gene:

functional genomics
.




Evolutionary biology:


Theories of evolution and

the origin of

species
.

M
olecular evolution
.

Origin of life and

the genetic code
.

E
cology:


The ecosystem

(why

can’t any

species


live in isolation
?)

Productivity (
from the sun
and nutrients to

biomass)
.

Decompisition

(from
biomass
to nutrients
)
.

Food and trophic levels

(
we are
all very similar
)
.


Ne
uroscience:


Psychobiology

of emotions
.

Psychobiology

of

memory
.