Team:USP-Brazil/Problem

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<p style="padding-top: 50px; margin-left: -40px;"><img src="https://static.igem.org/mediawiki/2013/f/f1/USPBrazilProblem.png" width="246" height="101" alt="Problem" /></p>
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<h4>The Challenge</h4>
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<p> The consumption of alcoholic beverages is part of global culture. Many places in the world have native alcoholic drink, which consumption is related to local events, parties and festivals [3]. Today, the global market of alcoholic drinks generates over a trillion dollars yearly [1]. </p>
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<p>Even though the consumption is large, most alcoholic beverages are not included as a formal share of the market. The  commercialization of “non-standard”, and the problem of adulterated beverages, despite both being neglected topics, is an increasing risk in leisure events. An estimate by the World Health Organization (WHO) suggests that about 30% of the alcoholic drinks sold in the world are not registered, reaching two-thirds of all the consumption in India, and shocking 90% in East Africa [2][3]. It is a global issue that has raised the concern of international organizations such as <a href="http://www.icap.org" target="_blank">ICAP</a> and <a href="http://www.amphoraproject.net" target="_blank">Amphora Project</a>.</p>
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<p>The commercialization of &#8220;non-standard&#8221;, or simply adulterated beverage, despite being a neglected topic in Brazil, is an increasing risk in leisure events [15]. An estimate by the World Health Organization (WHO) suggests that about 30% of the alcoholic drinks sold in the world are not registered, reaching two-thirds of all the consumption in India, and shocking 90% in East Africa [15]. It is a global issue that has raised the concern of international organizations as <a href="http://www.icap.org" target="_blank">ICAP</a> and <a href="http://www.amphoraproject.net" target="_blank">Amphora Project</a>. Methanol is one of the most common contaminants found in alcoholic drinks, able to provoke serious health disturbances, from metabolic acidosis to neurological problems. In large-fscale, a cheap and efficient methanol detector would undoubtedly help save millions of lives.</p>
 
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<p>Detection of chemical compounds has always been one of the goals of Chemistry. In spite of all the progress allowed by technological developments in analytical chemistry, like gas chromatography [1] and ionization flame detection [2], such a task remains largely unreachable for those without access to highly specific equipment. That is the case of methanol contamination in &#8220;non-standard&#8221; (adulterated) beverage, where one has to opt between heavy machinery or purely chemical methods&#8212;that is, those without the specific equipment - that are usually very elaborate and expensive [3]. It is also possible to use biochemical sensors [4], but those also lack accessibility, due to the usage of enzymes, as well as other expensive reagents, such as ubiquinone or luminol.</p>
 
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<p>The greatest advantage of a biochemical sensor could be, in addition to accessibility and easy use, its production: once the microorganism that has been modified to act as a detector is built, cell culture growth is itself responsible for producing the detector. Not only that simplifies the process, but it also reduces its cost: the only expenses after the development and construction of the biosensor would be with culture media and with preparing the product (e.g. lyophilization).</p>
 
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<p class="figure"><img src="https://static.igem.org/mediawiki/2013/3/3c/USPBrazilGraficoBebidas.png" width="433" height="295" style="border: none;" /><br /><b>Figure 1:</b> Infographic showing the proportion between registered and unregistered drinks around the world.</p>
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<h4> The Solution</h4>
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<p>Noncommercial beverages represent a serious risk to global public health, essentially due to the possible presence of drink contaminants that might lead to intoxications, and even to death. The mostly found contaminants are methanol, long chain alcohols, acetaldehyde, ethyl carbamate (possibly a carcinogen [4]) and metallic ions, such as copper and lead [5] [6] [7]. Among all contaminants, methanol regularly causes cases of severe symptoms and death, which appear often in the news. Our team decided to explore the possibility of detecting methanol in alcoholic drinks through engineered microorganisms, aiming to create a method that is not only accessible, but cheap and scalable, to analyse non-registered alcoholic beverages. In the process, we also intended to generate useful BioBricks for the Registry of Parts.
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<p>The organism to be modified to work as a detector is the yeast <i>Pichia pastoris</i>, which is an interesting choice due to its methylotrophic metabolism [5]&#8212;in other words, it uses methanol as a carbon source. <i>P. pastoris</i> is commonly used in the production of recombinant proteins [6], mainly due to its populational characteristics, such as growth rate and cell density, which make cell suspensions paste-dense [7], and to its methanol-responsive promoter, PAOX1. This promoter could be part of a genetic circuit that would respond to the presence of methanol by regulating the transcription of a reporter gene, responsible for indicating the presence of methanol by colouring the suspension (Figure 1).</p>
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<img src="https://static.igem.org/mediawiki/2013/1/15/USPBrasilCultivo.jpg" width="640" height="395" /><br />
 
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<b>Figure 1:</b> Fluorescent proteins expressed in an E.coli suspension. Respectively, amilCP BBa_K592009 (blue), amilGFP BBa_K592010 (yellow) and RFP BBa_E1010 (red).
 
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<p>PAOX1 is a strong promoter which can be controlled by simple changes in its carbon source [8], and is the most common choice for expression of heterologous proteins in <i>P. pastoris</i>, having a naturally elevated expression rate, of circa 5% of the RNA and 30% of total protein production [11]. Here, the chosen reporter was RFP (Red Fluorescent Protein).</p>
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<p class="figure"><img src="https://static.igem.org/mediawiki/2013/5/59/CHALLENGERecortesdeNoticias.png" width="450" height="333" style="border: none;" /><br /><b>Figure 2:</b> News showing the current problem relevance. </p>
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<p>The challenge in the building of this sensor was the regulation of PAOX1. This promoter is prone to a strong catabolic repression [12] by hexoses and ethanol&#8212;the main component of alcoholic beverages. Ethanol is also involved in the degradation of peroxisomes, cellular compartments where <i>P. pastoris</i> realizes the metabolism of methanol. This aspect is actually interesting to our application, since it means methanol will not be degraded as fast as it would, in the absence of ethanol. Therefore, methanol would stay for longer in the cell, being able to activate PAOX1.
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<h4>The Detector</h4>
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<h4>A Neglected Problem</h4>
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<ul>
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<p>Methanol can cause serious health disturbances, from metabolic acidosis to neurological problems [5]. According to the WHO, 100% of the pneumothorax and hemotorax traumas without an open wound (<a href="http://en.wikipedia.org/wiki/List_of_ICD-9_codes_800%E2%80%93999:_injury_and_poisoning" target="_blank">ICD 860.0 and ICD 860.2</a>) related to consuming alcoholic drinks are attributed to intoxication by methanol/ethanol [2]. The main victims of those injuries are people from low and middle-income countries, with economic conditions that do not allow access to legalized quality products [7], and yet there has not been any relevant investment, neither from governments nor from the private sector, on developing tools to help solving the problem of intoxication by methanol [8][9].
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<li>Como imaginamos o produto final em termos de relação com o usuário, e seu impacto pra resolver o problema</li>
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</p>
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<li>será elaborado um protocolo de tratamento a ser seguido pelo usuário, que consistirá em tratamento químico com hipoclorito e posterior retorno do produto pra tratamento adicional.</li>
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<p>A methanol detector would also be very useful to a second group of noncommercial drink producers: people from countries either developed or in development that take part in organized online communities, sharing methods of beverage production [10]. Unlike the previous case, their production is small-scale and solely for own consumption, and despite having access to specialized knowledge, often the producer's methods are quite rudimentary, lacking basic safety precautions [11][12][13]. A cheap and user-friendly methanol detector could improve the quality of the production of those home distilleries, reducing the risk of intoxication by methanol.</p>
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<h4>References</h4>
 
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<p>References of ProjetoOlimpiada-PessoaJr.</p>
 
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<p style="text-align:right;"><a href="https://2013.igem.org/Team:USP-Brazil/Project">See the solution&#8230;</a></p>
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<p style="text-align:right;"><a href="https://2013.igem.org/Team:USP-Brazil/Solution">See the solution <i class="icon-circle-arrow-right"></i></a></p>
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<h4 style="color:grey;">References</h4>
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<p style="color:grey;">
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[1] MarketLine. Alcoholic Drinks: Global Industry Guide. 2012.
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http://store.marketline.com/Product/alcoholic_drinks_global_industry_guide?productid=ML00004-125 <br></br>
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[2] World Health Organization (WHO). Global status report on alcohol 2004. Geneva: Author (2004).
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http://www.who.int/substance_abuse/publications/global_status_report_2004_overview.pdf<br></br>
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[3] World Health Organization (WHO). Global status report on alcohol and health. Geneva: Author (2011).
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http://www.who.int/substance_abuse/publications/global_alcohol_report/msbgsruprofiles.pdf<br></br>
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[4] B Zimmerli and J Schalatter. Ethyl carbamate: analytical methodology, occurrence, formation, biological activity and risk assessment. Mutation Research, 259: 325-350 (1991).<br></br>
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[5] DW Lachenmeier, J Rehm and G Gmel. Surrogate Alcohol: What Do We Know and Where Do We Go?. Alcoholism: Clinical and Experimental Research, vol. 31, nº 10 (2007).<br></br>
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[6] DW Lachenmeier et al. Is contaminated unrecoredd alcohol a health problem in the European Union? A review of existing and methodological outline for future studies. Addiction (Society for the Study of Addiction), vol. 106: 20-30 (2011).<br></br>
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[7] International Center for Alcohol Policies (ICAP). Producers, Sellers, and Drinkers: Studies of Noncomercial Alcohol in Nine Countries [Monograph]. Washington, DC (2012).<br></br>
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[8] International Center for Alcohol Policies (ICAP). Noncommercial Alcohol in Three Regions. Washington, DC, Review 3 (2008).<br></br>
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[9] CM Morel. Neglected diseases: under-funded research and inadequate health interventions. EMBO reports, vol. 4 (2003). <br></br>
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[10] http://homedistiller.org/forum/ <br></br>
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[11] http://homedistiller.org/intro/methanol/methanol <br></br>
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[12] http://homedistiller.org/distill/dtw/toss <br></br>
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[13] KA Berglund. Artisan Distilling - A Guide for Small Distilleries. Edition 1.0.0 (2004).
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http://distillery-yeast.com/free-pdf-documents/ </p>
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Latest revision as of 00:37, 28 September 2013

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Problem

The consumption of alcoholic beverages is part of global culture. Many places in the world have native alcoholic drink, which consumption is related to local events, parties and festivals [3]. Today, the global market of alcoholic drinks generates over a trillion dollars yearly [1].

Even though the consumption is large, most alcoholic beverages are not included as a formal share of the market. The commercialization of “non-standard”, and the problem of adulterated beverages, despite both being neglected topics, is an increasing risk in leisure events. An estimate by the World Health Organization (WHO) suggests that about 30% of the alcoholic drinks sold in the world are not registered, reaching two-thirds of all the consumption in India, and shocking 90% in East Africa [2][3]. It is a global issue that has raised the concern of international organizations such as ICAP and Amphora Project.


Figure 1: Infographic showing the proportion between registered and unregistered drinks around the world.

Noncommercial beverages represent a serious risk to global public health, essentially due to the possible presence of drink contaminants that might lead to intoxications, and even to death. The mostly found contaminants are methanol, long chain alcohols, acetaldehyde, ethyl carbamate (possibly a carcinogen [4]) and metallic ions, such as copper and lead [5] [6] [7]. Among all contaminants, methanol regularly causes cases of severe symptoms and death, which appear often in the news. Our team decided to explore the possibility of detecting methanol in alcoholic drinks through engineered microorganisms, aiming to create a method that is not only accessible, but cheap and scalable, to analyse non-registered alcoholic beverages. In the process, we also intended to generate useful BioBricks for the Registry of Parts.


Figure 2: News showing the current problem relevance.

A Neglected Problem

Methanol can cause serious health disturbances, from metabolic acidosis to neurological problems [5]. According to the WHO, 100% of the pneumothorax and hemotorax traumas without an open wound (ICD 860.0 and ICD 860.2) related to consuming alcoholic drinks are attributed to intoxication by methanol/ethanol [2]. The main victims of those injuries are people from low and middle-income countries, with economic conditions that do not allow access to legalized quality products [7], and yet there has not been any relevant investment, neither from governments nor from the private sector, on developing tools to help solving the problem of intoxication by methanol [8][9].

A methanol detector would also be very useful to a second group of noncommercial drink producers: people from countries either developed or in development that take part in organized online communities, sharing methods of beverage production [10]. Unlike the previous case, their production is small-scale and solely for own consumption, and despite having access to specialized knowledge, often the producer's methods are quite rudimentary, lacking basic safety precautions [11][12][13]. A cheap and user-friendly methanol detector could improve the quality of the production of those home distilleries, reducing the risk of intoxication by methanol.

See the solution

References

[1] MarketLine. Alcoholic Drinks: Global Industry Guide. 2012. http://store.marketline.com/Product/alcoholic_drinks_global_industry_guide?productid=ML00004-125

[2] World Health Organization (WHO). Global status report on alcohol 2004. Geneva: Author (2004). http://www.who.int/substance_abuse/publications/global_status_report_2004_overview.pdf

[3] World Health Organization (WHO). Global status report on alcohol and health. Geneva: Author (2011). http://www.who.int/substance_abuse/publications/global_alcohol_report/msbgsruprofiles.pdf

[4] B Zimmerli and J Schalatter. Ethyl carbamate: analytical methodology, occurrence, formation, biological activity and risk assessment. Mutation Research, 259: 325-350 (1991).

[5] DW Lachenmeier, J Rehm and G Gmel. Surrogate Alcohol: What Do We Know and Where Do We Go?. Alcoholism: Clinical and Experimental Research, vol. 31, nº 10 (2007).

[6] DW Lachenmeier et al. Is contaminated unrecoredd alcohol a health problem in the European Union? A review of existing and methodological outline for future studies. Addiction (Society for the Study of Addiction), vol. 106: 20-30 (2011).

[7] International Center for Alcohol Policies (ICAP). Producers, Sellers, and Drinkers: Studies of Noncomercial Alcohol in Nine Countries [Monograph]. Washington, DC (2012).

[8] International Center for Alcohol Policies (ICAP). Noncommercial Alcohol in Three Regions. Washington, DC, Review 3 (2008).

[9] CM Morel. Neglected diseases: under-funded research and inadequate health interventions. EMBO reports, vol. 4 (2003).

[10] http://homedistiller.org/forum/

[11] http://homedistiller.org/intro/methanol/methanol

[12] http://homedistiller.org/distill/dtw/toss

[13] KA Berglund. Artisan Distilling - A Guide for Small Distilleries. Edition 1.0.0 (2004). http://distillery-yeast.com/free-pdf-documents/

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