The Stubborn Life of PCBs

Tania Rabesandratana

What is the link between hermaphrodite polar bears in Norway, mass poisoning in Japan, and
GM tobacco in Prague? PCBs, a family of ubiquitous toxic molecules that have been banned a
long time ago, but still cause silent damage.

In the late 1990s, scientists discovered hermaphrodite polar bears near the island of Svalbard, in
Norway. Genetically, they were female, but had little penises in front of their vagina. Some
scientists put the blame on a family of man-made pollutants called Polychlorinated Biphenyls, or
PCBs. These molecules could mimic sexual hormones, hence messing with the bears’ sexual
organs.

PCBs have been manufactured and used since 1929 in many countries, e.g. under the market
name “Apirolio” in Italy. Because of their isolating properties, they became common ingredients
in many products, such as electrical transformers or paint. Unfortunately, PCBs are highly toxic.
In 1968 in Japan, kilos of rice bran oil were contaminated by the oil factory’s machines, causing
a PCB mass poisoning; thousands of people suffered from skin problems, menstrual troubles or a
weak immune system. Three decades later, in Belgium, poultry and pigs contaminated with
dioxin caused a memorable outcry. The public generally remembers this food crisis as “the
dioxin scandal” – but toxic PCBs were added to the feed mix, too. Animal studies confirmed that
PCBs are highly toxic for the liver and can cause cancer, or damage the reproductive and
immune systems. Most countries stopped producing them in the 1970s.

What does that have to do with hermaphrodite polar bears? Well, PCBs are very resistant and
can travel through water or air across thousands of kilometres, far away from polluting fabrics,
and decades after PCBs disappeared from the market. The water that reaches the Arctic has
accumulated PCBs along its way and the plankton is contaminated, too. Fish eat plankton, seals
eat fish, and polar bears eat seals… At every step of this simple food chain, PCBs accumulate in
the fat tissues to reach disruptive levels.

In 2001, the Stockholm Convention on Persistent Organic Pollutants blacklisted PCBs forever.
This international treaty requires that signatories do their best to prevent further damage.
Chemists and biologists are busy trying to clean up the mess, but PCBs are tough to get rid of.

Searching the hideout

PCBs have no particular smell or taste, and are too small to see with the naked eye. So how do
we know that they are here at all? Enter Kamila Kalachova, PhD student at the Institute of
Chemical Technology Prague. Kalachova is part of a sprawling EU-funded research project
called CONffIDENCE. Researchers in 10 countries are joining forces to establish cheap, reliable
methods to track contaminants in various types of food.
Kalachova hunts down PCBs in fish, such as smoked trout. From the initial fish sample, she
extracts a clear liquid and finds out if it contains PCBs, and how much. Part of her bench work
looks like kitchen experiments – adding a bit more of this solvent? Shaking the mixture a bit
longer? After about six months of trial and errors, Kalachova perfected a protocol that can now
be used routinely in her lab or in another. Her fast-track method can detect several types of toxic
molecules in one go and in a few hours, whilst the conventional method needed a few days.

Benches and corridors are bustling with dozens of young chemists searching fish, breakfast
cereal or beer for dozens of harmful contaminants. Yet, working in food safety research has not
affected Kalachova’s diet and habits. “I look at the ingredients, but I eat everything,” she says
confidently. Organic chemistry will keep inventing new contaminants, and analytical methods
can only do their best to keep pace. “If they do something that is free (of one contaminant), one
or two years later they’ll find something else. Everything is contaminated, anyway,” Kalachova
asserts. She does not seem worried, though: “the key is to eat small amounts of everything.”

Killing the villains

Three blocks down from Kalachova’s office, at the Czech Academy of Sciences, Jan Rezek
strives to eliminate PCBs from the surface of the Earth. The brutal, traditional method is burning:
at around 1000°C, PCBs are destroyed. Unfortunately, burning is unpractical and the by-products
themselves can be toxic. The newer trend is bioremediation, i.e. finding plants and microbes that
could do the job. It would be cheaper than digging out tons of earth to set them on fire, but more
delicate.

Rezek cultivated tobacco in the lab to confirm that plant tissues can take up PCBs and use them
for their own growth. He also exposes himself to toxic PCBs in dumpsites, looking for real-life
plants who can grow on contaminated soil, and searching for the best cleaning species. “Of
course it’s dangerous, but someone has to do it,” Rezek says simply. Then comes meticulous
detective work to reconstruct the chain of chemical reactions: understand how plants degrade
PCBs and what they turn them into.

Scientists hunt not only for plants who can degrade PCBs, but also for micro-organisms who live
comfortably in symbiosis around those plants’ roots. “We know various micro-organisms who
can (grow on PCB) in the lab, but under most natural conditions, they don’t survive well,” Rezek
explains. The metagenomic approach allows biologists to screen all genes of all bacteria living
around a plant’s roots at once. The big idea is to identify the genes responsible for the
degradation, and use them to create transgenic plants. Both transgenic plants and microbes would
work together to clean up the soil.

However, in the EU, strict regulations mean that GM plants hardly ever leave the lab. Such
transgenic tobacco could only be planted in the wild after a major change of policy. A fortiori,
we wouldn’t ever be able to sow such cleaning plants on the Norwegian ice floes, in an attempt
to save the next generations of polar bears from weird sexual symptoms.
Bohemian quest for bio remedy

Grażyna Zawada

How to harness bacteria and plants to work for people? Bohemian scientists are working on
improving cheap and ecological method of bacterial and plant degradation of toxic compounds
called polychlorinated biphenyls (PCBs)

- Our research can bring revolution in knowledge of total connections between microorganisms,
although its main task is bioremediation, achieved especially by cooperation between plants and
microorganisms – says Prof. Tomas Macek from Institute of Chemical Technology (VSCHT) in
Prague, Czech Republic. He and his team are working in a project called METAGENOM trying
to develop an easy and cheap way to degrade pollutants, mainly polychlorinated biphenyls
(PCBs).
These are organic compounds invented in late 1920’s and used widely in industry, especially as
dielectric fluids and coolants. Since their invention, ca. 1.5 million tons of PCBs have been
produced all over the world, even though they were known to be somewhat toxic. It was when
emaciated seabird corpses with very high PCB body burdens were washed up on beaches in the
1970s, that PCB toxicity was ultimately noticed. Further research showed that in highly
contaminated areas, both humans and animals were suffering mostly from skin conditions,
mainly cloracne, liver damage and deterioration of overall immune response. In addition,
although carcinogenic effects have not been explicitly confirmed, they are also suspected. PCBs
were officially banned from production in 1970s in most Western countries, however due to the
rule of the communist regime members of the Eastern Block member states, such as former
Czechoslovakia or Poland, did not stop PCB production and usage until several years later. In the
meantime ca. one third of PCBs had penetrated the environment by fires or leaks from faulty or
improperly stored equipment.
- PCBs volatilize very easily and migrate through the atmosphere along with air masses,
condensing in cold areas of the world, especially in North and South Poles. In animals, PCBs are
stored in fatty tissues and can accumulate in the food chain. The level of PCB in systems
increase matches the food chain dependence, with predators and people being most contaminated
– Ondrej Uhlik, PhD, adding that there even might be a correlation between PCB concentration
and some polar bears being born hermaphroditic.
- Since fats are the most representative for PCB concentration a test was carried out using butter
from each country of the world, as a representative for a large environment area – says Prof.
Macek. - New Zealand turned out to have least amounts of PCBs and Czech Republic had the
highest. Even our agriculture ministers’ blood was shown to have the highest level of PCBs of all
respective EU officials – says Macek.
Traditional method of destroying PCBs is e.g. burning, but the process is costly and sometimes
brings more environmental disturbance than the pollution itself. There is however another way:
- Using plants that are able to survive toxicity from soil is a cheap, solar driven phytoremediation
technique– says Jan Rezek, PhD, responsible for research on PCB metabolites in plants.
Plants have earned a name of green liver of the Earth, as they take in pollutants with water, then
detoxify them and use the compounds for storage or building their tissues. Soil reclamation can
be done by cutting the plants, drying them and sending to be burned, which is much cheaper than
other methods.
- The point is to select plants that deal well with PCBs – says Rezek. – My research is focused
on searching for plant species that take in as much PCB as possible, and done by extracting
metabolism compounds by gas chromatography and mass spectrometry. I select potential
candidates from plants growing in PCB dumpsites –says Rezek.
Another way of neutralizing PCBs is using bacteria that metabolize biphenyl. The end result of
this chemical process is just water and CO2 in case of biphenyl, structural analogue of PCBs. But
if so, why is there so much of it still in the environment? – Some more complex PCBs require
long lasting processes to degrade, and degradation of the more simple ones is not as efficient
under natural conditions as it is in the laboratory, as in most cases if efficiently degrading
bacteria are added to contaminated environment they die, being pushed out by original bacteria.
My part of research is to identify natural working microflora and find proper conditions for
indigenous bacteria to work – says Uhlik, PhD, responsible for bacteria research for
METAGENOM project.
Finding really active microorganisms is done by stable isotope probing, a technique used only by
a few laboratories all over the world apart from VSCHT in Prague. Bacteria in PCB
contaminated soil are provided labeled or heavy carbon biphenyl and then observe how they
utilize it and also integrate the label in their DNA when they multiply. After due time the
metagenome, meaning all DNA sets of whole bacteria population from certain sample is
centrifuged from the sample and Dr. Uhlik and his team can isolate heavy DNA of organisms
that have utilized biphenyl, clone it and thus, by multigenomic sequencing identify their name
and specific properties in degrading particular PCB types.
No more than 5 years ago scientists could work with identifiable, laboratory cultivable bacteria
only, knowing only ca. 30 percent of bacterial DNA, but not being able to recognize function of
particular genes. - By this method we are able to recognize the role of every gene, contributing
also to the study of all bacterial metagenome of the Earth and of terragenome, genome set of all
living organisms on Earth. It is a tremendous task, but it will slowly contribute to identifying all
existing genes and can be further used in biotechnology – says Prof. Macek.
It takes a warrior to fight toxins

Grażyna Zawada

When I first saw Jan Rezek I thought ‘Is that a scientist or a rock star’? Indeed, with his huge
biceps, over a dozen earrings and a small plate at the back of a skin-shaven skull he contradict
stereotypes about scholars
Jan Rezek, PhD is currently doing biochemical research focused on removal of toxic substances
called PCBs from the environment. He works in Joint laboratory of IOCB (Czech Academy of
Sciences) and Institute of Chemical Technology, Prague. His interests have shaped not only his
appearance, but also personality.
Grażyna Zawada: Judging by your looks, you must have an unusual hobby.
Jan Rezek, PhD: Well, nowadays my main hobby is Musado, a martial art of Korean origin. It
is basically a very efficient self defense street fighting style; there are no special rules, but it is all
about defending with minimum effort and maximum results. It was developed by a German who
went to Korea to learn old Korean martial art and then modernized it by removing elements
useless in street fights, e.g. acrobatics, and nowadays even Czech army uses it as their combat
style.

Have you ever had to defend yourself in the street?

I’d say in some situations it’s better just to give the money to the guy and save your life, or just
run away if there’s a chance, as fighting can always end up with losing a life. But I think people
avoid fighting me when they see me, even in really dangerous places.
Any other hobbies besides Musado?
Some time ago I played bass guitar in a band. I also like to travel when I have the opportunity.
Where have you been so far?
I spent a year in Colombia, where I met my wife, Alba. I’ve also been to Africa and I hitch-hiked
all over Europe.
Well, why becoming a scientist then?
I’ve always been attracted to nature, and when I was a child my father would always explain
things to me as both of my parents are teachers. I guess this combination was what shaped my
future career. When you are a scientist you in a way always remain a child, because you
constantly learn something new and discover things.
What do you plan to do in next ten years?
There are basically two options: I’m here in Czech Republic or I’m in Colombia, as Alba and I
want to stay close to one of our grandparents who would help us with babysitting (smiles).
Currently, if my wife decides on a place to complete her specialization in medicine I will try to
follow her and find an appropriate place for a PostDoc there. The research I’m doing now is a
kind of a second line fight, as PCBs do not kill you instantly. Perhaps in the future I’ll move to
the front line of combat.
What do you mean?
In Colombia I worked in a lab connected to tropical diseases, where I worked e.g. with multi-
resistant tuberculosis. In that type of research you have to work fast to determinate specific
bacteria or viruses, and if not, the infected men will die in several days. Perhaps in the future I
would like to do something similar, even complementing my wife in her medical work. That’s a
first line combat.
Revealing a person, revealing a dream

Elena Vulpe

When was last time you helped your mother/grandmother prepare Christmas Dinner? Spending
few days in the Chobotix Laboratory I felt like a child that did some magic in Granny’s kitchen,
the only difference was that the final product wasn’t for eating, at least not at this stage. But first
what is Chobotix and who is the Chef?

The Man: A Young Czech Scientist that leads an enthusiastic team to the chemical robot’s era.
Dr. Frantisek Stepanek has gained respect from his colleagues due to his multitasking ability. He
is Senior Lecturer at the Imperial College of London since 2006, has received several awards
including the prestigious Junior Moulton Medal from the Institution of Chemical Engineering,
the Philip Leverhulme Prize for Engineering and the Friedrich Wilhelm Bessel Award. He has
four children and likes to spend his free time with his team and to go skiing with family. As one
of his colleagues said, he seems to have discovered a way to make the day 36 hours long because
he manages to be everywhere where he is needed.

The Dream: Chobotix is the acronym for Chemical Processing by Swarm Robotics and its main
idea is to create chemical systems that will be able to target a specific region in an environment
and selectively fix problems in that specific region. It should be a micrometer scale chemical
laboratory that will work independently and act locally in response to some external factors. It is
a FP7 project that will be financed from 2008 until 2013.

How did they meet?: It all started with Science Fiction and Dr. Stepanek’s post-doctoral
experience at Unilever’s R&D Center. There he was working with solid particles that were too
rigid and didn’t leave much room for creativity. So he thought about some particles that can
change their shape in response to different parameters such as: temperature, concentration, pH
(acidity or alkalinity). More than that, he thought that it would be interesting if particles could
transport some active substances and deliver them where they are needed.

And here starts the magic!

As visitors in the ICT Prague laboratories we followed the same steps that were made by the
scientists. First of all they wanted to obtain solid particles which could act as carriers. For that
we used a Teflon glass that we filled with few milliliters of octylamine and stirred it on a special
magnetic hotplate at normal temperature. After few minutes we added a few milliliters of
tetraethyl silicate and stirred it for another few minutes. As spices we added a fluorescent dye
(FITC*) which permits the visualization of the particles under a microscope. Then we added
nitric acid which acts as a catalyst. We left the mixture for 10 minutes on the plate and observed
the formation of a turbid solution. We washed this solution with water and watched the
deposition of some little particles. When it is ready (you have the turbid solution with some
precipitate) you put it in oven for few hours at 550⁰ C. When this step is done you can observe
your particles on a microscope so you can describe their surface properties. The particles will
look like the ones in the images below.

SEM images of silica particles Particles on optical microscope

The second step is to fill silica particles with a substance that will be able to change their
properties under the influence of external factors. For this scientists have chosen a polymer for
encapsulation, the most suitable were alginate and N-isopropylacrylamide (PNIPAM). Both are
hydrogels with interesting properties, alginate is widely distributed in cell walls of brown algae
and is used for detoxifying from heavy metal poisoning. PNIPAM has a beautiful property of
shrinking at higher temperatures and expanding back when the temperature is reduced. The
question was whether the particles filled with polymer could be preserved in a dry state and
without losing their ability to change volume in response to temperature variations.
Experimentally it was shown that particles preserve their properties and after re-dispersion their
internal structure and properties were restored.

Shrinking / swelling behavior of PNIPAM capsules coated with silica nanoparticles

Simultaneously iron nanoparticles were synthesized in the laboratory. They can induce local
heating under the influence of external factors. This property of iron nanoparticles will be used
for transferring the heat to the hydrogel and under this effect “chobots” will release active
substances or drugs preserved inside capsule only at the injured parts, organs or tissues. Other
experiments have to be done for solving all the problems that could appear during the process.
The mobility problem is the next concern. When looking for some models Dr. Stepanek’s team
found the answer in nature. It was observed that some bacteria under lack of nutrients form a
pluricellular body. The mechanism of this process is not precisely known but there are
microbiology research centers which are trying to establish it.
That is a brief presentation of the steps the team made until this moment, during these two years
articles were published in materials science journals about their work and important steps in
creating a prototype of chemical robots were made. As Dr. Stepanek said, they are at the
beginning and it’s important that the European Council and ICT Prague finance such
revolutionary projects and offer the possibility for independent scientist to apply their ideas in
research.

Textbox:

FITC- is a fluorescent compound that in water has yellow color and under UV-light is green. It is
used for observing particles under the microscope which are usually not seen with the naked eye.

Hydrogel-is a network of polymer chains that are insoluble in water and posses a high level of
flexibility. Hydrogels are highly absorbent and under the increase of temperature they shrink due
to loss of the absorbed water, the process is reversible.
A new weapon in the fight against MRSA

Elles Lalieu

Czech researchers are developing special layers as a weapon in the fight against ‘the hospital
bug’ MRSA. These layers are now being tested in the laboratory. In the future the layers must
prevent the spread of harmful bacteria in both stables and hospitals.

Have you ever heard of MRSA? This bacterium is causing major health problems in hospitals
throughout Europe. And there was no way to fight it, until now. Czech researchers are
developing solutions that can destroy harmful bacteria in no time. You just have to spray it on
the walls and turn on ultraviolet (UV) light. Two of the researchers show us exactly how it
works.

Staphylococcus aureus is a common bacterium frequently found in your nose and on your skin. It
can grow fast on surfaces like tiles, floors and windows. When your immune system is weak, for
example when you are sick or just had surgery, this bacterium can cause nasty infections. And
once you are infected it is hard to find a good cure. Since the introduction of penicillin in 1943
almost all strains of Staphylococcus aureus became resistant to widely used antibiotics. MRSA,
also known as ‘the hospital bug’, is the most common form of these resistant bacteria.

In 2005 researchers discovered an animal derived clone of MRSA that causes disease in pigs and
piglets, but also in humans. This clone is used in the European FP7 research project PILGRIM to
study MRSA both in veterinary settings and in public health. Josef Krýsa and Jiří Zita from the
Institute of Chemical Technology, Prague (ICT Prague) are participating in PILGRIM. In the
battle against MRSA they are developing surfaces coated with the mineral titanium dioxide
(TiO2). Their hope is that, in the future, walls and floors of hospitals can be cleaned with a
solution of TiO2 that destroys harmful bacteria in no time when illuminated with UV-light.

Preparing layers with the ‘dipping’ device

“TiO2 is a photocatalyst, which means it uses UV-light to speed up a chemical reaction between
water and oxygen.” Zita explains. “Reactive particles, or free radicals, are produced as a result.
These particles can attack bacteria by first destroying their cell wall and later also their cell
membrane.” Several different solutions, containing TiO 2 particles, are prepared and tested in the
laboratory of ICT Prague at this moment. Zita shows two of them: an orange solution that has to
be heated before it can be used and a transparent solution that can be used directly on surfaces.
“The transparent solution would be ideal to use, but it is in our laboratory for only a few months
now. So it still needs extensive testing.”
The reactivity of solutions is tested under UV-light (behind), but also under visible light (at the forefront). The range of
visible light in the solar spectrum is much greater than that of UV-light. Therefore, a TiO 2 layer that reacts under visible
light is more practical.

With a mechanical ‘dipping’ device, the solutions are put on small glass slides. In this way TiO 2
layers of different thickness are produced. Zita already discovered that a thin layer of TiO 2
solution is the most stable: “It is difficult to make scratches on a thin layer or remove complete
parts of it from the glass slide. But on the other hand, the chemical reaction of a thick layer is
much stronger because it absorbs a higher amount of light. In other words, the activity of a thick
layer is higher. Finding of an optimal layer thickness is necessary.”

“The activity of the layers is tested by dipping the glass slide coated with TiO 2 into blue ink.”
Zita continues. “Under UV-light produced reactive particles attack the color pigment of the ink.
When the layer is active the ink will become transparent after just a few minutes of UV-
radiation. The ink of layers that are not active will stay blue, even after more than several hours
under UV-light.” Layers that pass the test of activity are transferred to the microbiology
laboratory of ICT Prague. Here it is investigated if those layers are really useful for the
destruction of living bacteria.

Don’t be concerned about reactive particles

Zita: “Our microbiology laboratory is not equipped to work with MRSA so the layers are tested
on Escherichia coli (E. coli), another bacterium.” E. coli grows really well on glass plates when
the right culture medium is provided. You can measure this growth by counting the bacteria
colonies from time to time. “At ICT Prague we build a special testing chamber to irradiate E.
coli bacteria, placed on TiO2 layers, with UV-light. If a layer is working well the number of
bacterial colonies should go down.” TiO2 layers that are successful in these test will be send to
the Royal Veterinary College in London. Here the layers are tested on living strains of MRSA.
The testing chamber at ICT Prague where E.coli bacteria, placed on TiO2 layers, can be irradiated with UV-light.

While all these laboratory tests are still in progress Krýsa and Zita already thought of the
practical implications of their cleaning method. Is it safe to clean hospitals using UV-light and
reactive particles? According to Zita it is. “To start the chemical reaction we use UVA-light with
a wavelength of 330 to 400 nanometers. That kind of UV-light is part of the sunlight that reaches
your skin every time you take a walk outside. It is no problem to turn on the UV-light, for say
half an hour, to clean a hospital room full of patients. Some people are concerned about the use
of reactive particles, but there is no need to be. These particles have a very short life time. Once
they have killed a bacterium, they are gone.”
Before we can benefit from the effects of TiO 2, pigs will. In collaboration with Danish
researchers, the ICT Prague will develop specially equipped stable boxes. The air in these stables
is constantly cleaned. Walls and floors will be covered with white paint containing sufficient
amounts of TiO2. During half of the day UV-lights will be on to remove MRSA out of the pig’s
environment. If this works Krýsa and Zita have developed a method that can not only keep
MRSA out of the stables, but also away from the farmers and even far from the hospitals.
WELTEMP Project helps to protect the environment

Elmas Işık Yenerer

Hydrogen as a 21st century cheap and reliable source of energy carrier can be produced by
electrolysis of water. This was a challenge but expensive way since 1927’s. WELTEMP opens
the gate of producing cheap hydrogen and gives a hope in terms of protecting the environment.

Hydrogen is a good energy carrier in order to provide a reliable, and clean source of power.It can
be produced from conventional fossil fuels, from biomass, and by electrolysis of water.
Electrolysis has been used for approximately 100 years for hydrogen production as a cleanist
way. The first large hydrogen production installation was founded by Norsk Hydro in 1927 in
Norway..
There is an experiments about water electrolysis in Instute Chemical Technology Prague ‘s
(ICTP) laboratories which I was invited for the workshop. I had an oportunity to work with
Prof.Karel Bouzek’s team. Bouzek is the head of the Departmant of Inorganic Tecnology at the
Instute of ICTP and Vice Dean of the Faculty of Chemical technology.His areas of interest are
full cells,water electrolysis,technical electrochemistry and electrochemical engineegiring with
particular focus on the treatment of waste drinking water.His team is ICTP’s representive on the
research group of the Joint Technology Initiative(JTI)on fuel cells and hydrogen(FCH) This team
is working on WELTEMPT (WATER ELECTROLYSIS AT ELEVATED TEMPERATURES )
Hydrogen by means of water electrolysers can be produced in several ways. When renewable
energy sources such as hydropower, windmills, solar cells, etc. are considered, electrolysis is the
practical way of converting the surplus electrical energy into chemical energy to be used when
the power is needed.
This way is the more secure and clean way of using hydrogen production.But the biggest barrier
to the hydrogen economy is the challenge of getting hydrogen economically to the point of use
like process of production, storage and transportation
The WELTEMP project funded by the European Union is a collaborative project under the
Seventh Framework Programme (FP7). The project involves partners Denmark, Norway, Czech
Republic, Switzerland and Italy and has started in 2008 and will go on till the end of 2010.
PEM (PEM electrolyser-membrane electrolysis (acidic pH))process is more cheap than alkaline
process
They are making a research on production of cheap hydrogen by water electryolsis.. Everything
is available in the future with hydrogen. For example In recent years some car companies have
released the passenger vehicles use hydrogen power. BMW (LH2), Renault (LH2) and ZEVCO
(CGH2); van-type vehicles of Daimler-Benz (CGH2), Hamburg Hydrogen Association (CGH2),
PSA (CGH2) and ZEVCO (CGH2) and city buses; Ansaldo (LH2 ), Daimler-Benz(CGH2).
These university researches are going to light our future with clean energy sources for industry,
areas of daily needs.
Basta un poco di zucchero…

Irene Gabrielli

Molti cuochi italiani si trasferiscono all’estero per esportare le tipicità nostrane, dai canederli ai
malloreddus. Altri, un po’ speciali, emigrano per preparare piatti altrettanto speciali. Zuccheri.
Che cucinati a dovere, possono prevenire infezioni da parte di alcuni dei virus più aggressivi per
l’uomo come Ebola, hiv o tbc. È il caso di Benedetta Bertolotti, trentenne cremonese, che dopo
la laurea in chimica industriale all’Università di Milano ha scelto un dottorato all’Istituto di
Tecnologie Chimiche di Praga (ITC) per lavorare al progetto Carmusys.

Cosa significa la parola Carmusys e quali sono le finalità di questo progetto?

Bertolotti: Carmusys è un progetto che riunisce enti di ricerca operanti in diversi ambiti e in
diversi paesi europei. Il nome è un acronimo inglese che in italiano suona più o meno come:
sistema di carboidrati polivalenti per studiare interazioni patogene con dc-sign. Il nome può
spaventare ma, in realtà, nasconde un concetto piuttosto semplice: si tratta di studiare le
interazioni della proteina dc-sign, solitamente presente in tutti gli esseri umani, con alcuni agenti
portatori di malattie, principalmente virus. Sappiamo che questa proteina si trova sulla parete di
particolari cellule del sistema immunitario: quando un virus penetra nell’organismo, si attacca ad
essa e la sfrutta per veicolare l’infezione dentro alla cellula, e da lì a tutto il resto del corpo. Il
progetto Carmusys cerca di costruire chimicamente delle molecole, nella fattispecie zuccheri,
che siano in grado di legarsi alla proteina prima e meglio di quanto non faccia il virus, in modo
da impedirne l’attacco e quindi l’infezione.

Qual è il suo ruolo all’interno del gruppo di ricerca?

Una volta che i colleghi del modelling hanno progettato al computer la struttura che sembra la
più adatta a legarsi a dc-sign, entro in gioco io per cercare di riprodurla nella realtà. Il mio lavoro
parte da uno zucchero, il mannosio, che in natura si trova abitualmente nelle palme o nelle
carrube e che già di per sé mostra un’alta affinità con dc-sign: attraverso una serie di reazioni che
possono essere anche venti o quaranta, ognuna delle quali può richiedere dai cinque minuti ai
cinque giorni, io cerco di dare allo zucchero la forma più adatta per renderlo ancora più
“appetibile”, irresistibile, per la nostra proteina. Nell’impostare queste reazioni mi affido a
esperienze note, riportate in letteratura, ma come succede in cucina, a volte le ricette devono
essere un po’ modificate a seconda di quello che uno ha a disposizione e di quello che vuole
ottenere. Capita che per una sequenza di quaranta reazioni mi trovi a dover consultare sessanta
“ricettari” diversi. Una volta riuscita a sintetizzare uno zucchero della struttura che cerchiamo,
toccherà ai laboratori di biologia e virologia testarne l’efficacia. Ad oggi il nostro gruppo ha già
trovato, per alcuni di questi zuccheri modificati, un buon effetto antivirale contro i virus
dell’HIV e quello dell’Ebola.
Quali sono gli enti coinvolti nel progetto Carmusys?
Si tratta di un consorzio di ricerca finanziato dalla Comunità Europea nell’ambito del VII
Programma Quadro, che coinvolge dieci centri di ricerca universitari e due privati provenienti da
sette paesi diversi: Spagna, Francia, Italia, Germania, Olanda, Regno Unito e, ovviamente,
Repubblica Ceca. Oltre a me ci sono altri quattro italiani: una è Anna Bernardi dell’Università di
Milano; gli altri sono due dottorandi e un PostDoc, anch’essi emigrati, in Olanda e in Spagna. È
un progetto triennale che si concluderà nel 2012, per il quale la Comunità Europea ha già erogato
i fondi, stabilito la tempistica e ora monitora sui risultati richiedendo frequenti questionari e
rapporti sullo stato di avanzamento della ricerca. Per questo ogni sei mesi ci ritroviamo tutti in
“gita” in uno dei laboratori coinvolti per discutere i progressi del lavoro, vedere cosa funziona ed
eventualmente correggere il tiro.
Cercare un dottorato all’estero: più un desiderio o una necessità?
Direi entrambi: già durante la tesi di laurea mi ero scontrata con i ben noti problemi
dell’Università italiana. La mancanza di finanziamenti, i tagli , il precariato… L’alternativa era
orientarsi al settore privato ma in questi anni anche le grandi multinazionali tendono a chiudere
molte delle loro sedi, ad accorpare tutto in pochi centri di ricerca grandi e a trasferirli in paesi
dove hanno maggiori sgravi fiscali e incentivi. Diciamo che “costretta” a cercare qualcosa mi
sono orientata verso un dottorato, perché rimane un titolo molto appetibile per un ricercatore, e al
di fuori dell’Italia dove spesso è più semplice trovare spazio o comunque ci sono maggiori
possibilità. Tra le tante domande inviate, mi hanno risposto dall’ITC di Praga per lavorare a
Carmusys: mi è sembrato subito molto interessante, il paese mi ispirava, e mi sono detta, perché
no?
Lei che ha lavorato in entrambi, quali differenze nota tra un laboratorio italiano e uno
ceco?
Per quella che è la mia esperienza direi che rispetto all’Italia, i miei attuali colleghi hanno, in
generale, un’età media più bassa; e questo vale sia per i ricercatori che per i professori. Per
quanto riguarda il laboratorio chimico, invece, non ci sono grandi differenze di strumentazione di
base. Però qui in Repubblica Ceca la ricerca gode di maggiori finanziamenti e quindi ci sono
meno problemi per ordinare gli ingredienti per le reazioni, si investe nell’acquisto di nuovi
apparecchi,… Questo è importante per un laboratorio perché consente di avere analisi più rapide
e più precise. Ma senza andare sulle grandi spese, basti pensare che quando lavoravo in Italia mi
sono dovuta comprare da sola il camice di laboratorio, mentre qui considerano equipaggiamento
base, non solo camice e pantaloni, ma anche le chiavette USB!
PER SCINDERE I GAS CI VUOLE IL FISICO. ANZI, IL CHIMICO

Sara Ficocelli

Il dottor Karel Friess parla un buon italiano e ci accoglie con gentilezza, scortandoci fino al
laboratorio n.137 dell'ICT un po' preoccupato che al ritorno non si riesca a trovare la via d'uscita.
"Ho lavorato al Cnr di Reggio Calabria per tre anni - spiega - lì c'è l'Institute on Membrane
Technology. Ricordo con nostalgia il mare e soprattutto il cibo, mi manca molto l'Italia". Il
ricercatore del dipartimento di Physical Chemistry dell'Institute of Chemical Technology di
Praga un anno fa è tornato nella capitale ceca per portare avanti il progetto DoubleNanoMem,
per il quale la UE ha stanziato più di 4 milioni di euro e che entro il 2011 dovrebbe portare a
capire come fare a scindere i gas, controllando il potere di assorbimento delle membrane che
fanno da filtro. La cucina calabrese non è riuscito a tenerlo lontano da un lavoro che racchiude le
conclusioni di dieci anni di studi, e che potrebbe ad esempio aiutare gli ospedali a purificare
l'aria dalle sostanze nocive invisibili, quelle che finora i normali sistemi di filtraggio non
riescono a bloccare. "Il nostro compito è quello di testare il potere di assorbimento delle
membrane ai polimeri, versatili per definizione ma proprio per questo aperte ad ogni possibile
reazione fisico-chimica. Una volta capito il margine di resistenza, potremo sviluppare una
membrana capace di intrappolare le microparticelle nocive presenti nell'aria, lasciando andare le
altre". Detta così sembra facile ma dando un'occhiata al laboratorio gestito da Friess e dal
professor Milan Šípek ci si accorge subito che in questo progetto di ricerca non c'è nulla di
semplice. A cominciare dal funzionamento dei macchinari, tutti progettati dagli stessi ricercatori
e da loro assemblati, dei capolavori del fai-da-te che dietro ad ogni bullone nascondono un
mondo di equilibri incomprensibili, per lo meno per noi digiuni della materia. Il ricercatore, che
ha da poco avuto il secondo figlio, ci mostra al pc le immagini dell'ultimo arrivato con un
briciolo di emozione, e poi ci spiega che la realizzazione di questo progetto è costituita da due
momenti fondamentali, uno di creazione delle membrane e uno di test di resistenza sulle stesse.
Tutto comincia la mattina alle 9 con una serie di esperimenti a catena che si ripetono per tutto il
giorno, conservando i dati prima a penna, su un registro tenuto con precisione, e poi su un
database elettronico. Per sperimentare la capacità di assorbimento delle membrane ai polimeri i
ricercatori hanno sviluppato un sistema di lavorazione dell'acqua attraverso il nitrogeno, un gas
liquido che la trasforma in ghiaccio in pochi secondi. Questo passaggio permette di lavorare
sull'aria privandola dell'ossigeno e dell'ossido di carbonio, e di capire fino a che punto le
membrane sono capaci di assorbire le sostanze che restano. "Per il momento i risultati sono
promettenti, tutto sta andando nel verso giusto - continua Friess - ma nel nostro lavoro è tutta
un'incognita, i dati possono riservare sorprese inaspettate. Personalmente, amo questo lavoro
perché mi permette di mettere alla prova i limiti della materia ogni giorno". Con lui lavora anche
Kristoff, un ragazzo di 23 anni che maneggia le apparecchiature del laboratorio con la destrezza
di un veterano, e Angelika, che si occupa della preparazione delle membrane. Il professor Šípek
ha ormai una certa età e supervisiona il lavoro con il sorriso bonario di un nonno con i nipoti.
"Ho dedicato la mia vita a questo tipo di ricerca - racconta - Se il progetto dovesse concludersi
con successo si potrebbe passare alla fase due, quella della trasformazione dei dati teorici in
qualcosa di pratico". La fase tre è quella più interessante dal punto di vista economico, perché
permetterebbe all'Università di Praga di ricevere altri fondi per ricerche più specifiche, magari
sulla base delle esigenze di mercato. Ospedali, scuole, industrie automobilistiche, altri laboratori
di ricerca: il panorama delle realtà che potrebbero essere interessate ad un sistema di filtraggio
dei gas ad altissimi precisione è infinito. Un po' come infinite sono le possibilità di
sperimentazione della materia, per dei chimici in gamba come quelli dell'ICT.
Watch out: dangerous pollutant lurks on our lives!

Zuzanna Kaczmarska

Dr Jan Rezek from Czech Republic warns that polar bears are exposed to a threat of giving birth
to offspring having both female and male sexual organs (also known as hermaphroditism). The
cause of this phenomenon is certain toxic chemical compounds, which accumulate in the animal
fat tissue, e.g. tissue of bears or fish. These animals, just like people, are the final link of the food
chain, therefore, they are subject to assimilation of dangerous compounds contained in food of
both animal and plant origins.
How have I come across dr Rezek? When I first saw this well-built guy with a characteristic eye-
catching hairstyle and with piercings in his ears, who was staring at workshop’s participants in a
bizarre manner, I couldn’t stop wondering who on earth he was, and what he was doing in a
chemical laboratory. This freshly defended PhD student of biochemistry at the Institute of
Organic Chemistry and Biochemistry in Prague, Czech Republic was one of our lecturers who
was supposed to introduce us to the world of modern chemistry and biology, which he focuses
on in his everyday work. He began our session by introducing himself. I was not surprised to
hear that in his free time he trained martial arts such as musalo. Moreover, he loves travelling. It
was when he was on an internship in Colombia that he met his wife Alba.
For a few years now dr Rezek has been engaged in scientific research whose purpose is to
identify the plants having capacity to decompose PCBs (PolyChlorinated Biphenyls), a
dangerous pollutant which can cause serious hazard to human health.
Commercial production of PCBs was firstly commenced in late 1920s in the US. Due to its poor
flammability they are good insulation material. They have been used widely as coolants,
lubricants in transformers and in other electrical equipment. With increasing knowledge of their
toxicity and ability to transfer to food chains and accumulate in living organisms, their
production ended in most countries in the 1970s and in 1984 in the former Czechslovakia (Rezek
et al. Chemosphere, 2007, 69, 1221-1227) . In particular, it was shown that people exposed to
these chemical substances had skin conditions problems, liver damage, lower immune response
and poor cognitive development. Additionally, babies born to women who ate PCB-
contaminated fish had problems with motor skills, short-term memory and also their immune
system was affected. Finally, the studies have shown that PCBs contribute to reproduction
problems, such as hermaphroditism.
PCBs are very stable compounds and do not degrade readily, therefore they need to
be annihilated with the use of special methods. Their destruction by chemical, thermal and
biochemical processes is extremely difficult and presents the risk of generating exceptionally
toxic derivatives. Intentional degradation as a treatment of unwanted PCBs generally requires
very high temperatures or catalysis (Mackova et al. Environ. Sci. Pollut. Res. 2009, 16, 817-
829).
Volatile PCBs evaporate and contaminate vast areas, even at very distant parts of the world, for
instance polar regions. In the environment they mainly accumulate in the hydrosphere. Despite
their hydrophobicity, the great amount of water in oceans can dissolve a fair amount of PCBs,
but they are also found in the organic fraction of soil and in living organisms.
Small organisms absorb PCBs in the water and are then eaten by small fish. Large fish eat
smaller fish, and in this way, the contaminants accumulate and become transferred up the food
chain. The top level predators such us walleyes, polar bears or humans can absorb great amounts
of these dangerous chemicals. Since PCBs are stored in fat, they are not easily digested and
cannot be removed from the body. Accumulated PCBs can cause many health problems
including, as mentioned above, hermaphroditism or failure in sexual organ development. For
these reasons, PCBs represent a global threat.
At present, the main method of PCB removal from contaminated environment is an expansive
incineration in high temperatures. With the aim of finding effective alternative approaches, dr
Jan Rezek is studying biological methods for PCBs removal from the soil. In his research he uses
the soil from dumpsite in South Bohemia, Czech Republic and tries to investigate the most
effective plants, which are able to absorb or/and transform the PCBs to the non toxic compounds.
“This method is called phytoremediation” – dr Rezek explains.
„First step is to pick up the plants which grow on the contaminated area” – says dr Rezek – “then
we extract all the compounds from them and analyze them by gas chromatography (GC) and
mass spectrometry (MS)”. The GC facility which dr Rezek employs in the research allows him to
separate the chemicals extracted from the plant. “It can be compared to the race in a tube which
are lined with the proper chemical compounds with special properties allowing them to catch and
slow down the competitors” – dr Rezek sums up. It means than the compounds can finish the
race in different time according to their properties. In other words, they can be separated. The
second step is to identify the participants of the race. In order to do this, dr Rezek analyzes their
masses by use of mass spectrometry method. “I am mainly interested in PCBs and their
metabolites. Each PCB has its own fragmentation pattern by which can be identified. The
fragmentation pattern is like a fingerprint of a molecule. You cannot mistake it for another." – dr
Rezek clarifies. The presence of the PCBs and their derivatives in a plant suggests that this
particular plant can be used for the detoxification of the contaminated soil. Moreover, these
plants support microorganisms which can also have the ability to degrade toxins.
However, at this very moment dr Rezek cannot be sure whether he will continue the research. He
and his Columbian wife are planning to move to Spain to live there. Time will show us who will
solve the problem of PCBs in the Czech Republic assumed it actually can be done whatsoever.
 Destruction of the plant cells by the liguid nitrogen.

Extraction of the compounds in the separator. How the gas chromatography looks like inside?
 Here we have samples ready to be ananlysed by (gas chromatography) GC and MS (mass
spectrometry).

Preparation for the experiment.

Let's pour a little bit of liguid nitrogen.
 Let's see if the compound have been extracted!
It is really important to destroy all
the plant cells to release all the compounds.

This machine allow us to separate and analyse the compounds from the plant.