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The Scientific Ravi 20 II

Expanding Horizons in theWondrous Journey of Life

Sciences

jluhallllllad AlifKI/(JII

The game started long ago when perhapshunting and gathering was the only idea offun for man. There was no internet andtherefore. no Facebook or Skype to assist insocializing. But man managed to sailthrough thick and thin. Trying to controlnature was a habit encoded within hisgenetic blueprint. After all. when everythingwas made subservient to man. bmins weregi fted along by God to harness thosecontrols. Dinosaurs couldn't have predictedthat their remains ,\'Ould be used to fuelmachines upon IIhich the world is running afell' million years later. Man has bredlivestock to become more productive anddocile than their wild counterparts. Heartificially enhanced the yield and quality ofcrops like maize and corn through selectivebreeding. There was always a longing thirstto control and understand the extremelycomplex systems of nature. but men like thislIere almost always unwelcomed. especiallyby those who delined the e.xisting order. Tocut a long story short. Bruno lI'as burnt alh'eand Galileo was tortured while many others"ho belonged to their creed lIere met lI'ithterrible fates.

Today "hen decades have passed since thefirst successful experiment of geneticengineering was done and many years sincethe first cloned sheep Dolly was born. thejourney is lar Irom being over. Some call itmeddling "ith polI'ers beyond our controland bioethics emerges to curb the ill-eflcctsof such intrusions. The turn of thismillennium saw the entire human genomegetting sequenced in the Iluman GenomeProject and a decade later the creation of

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lirst semi-synthetic cell was reported.Cloning was about photocopying the DNA.but making a synthetic cell that is sci I~replicating and fully functional "as aboutIITiting the genetic code in a process toimitate nature. It is still semi-syntheticthough. for it uses a genome printed by acomputer. but requires a fully functional cellminus its origi\wl DI\!A in order to be able togrow and divide because despite all thescientific advancements. man has not yetbeen able to produce artificial protoplasm.Proponents of synthetic biology say. II ithever increasing number of people. thedemand It)r fl)od. tllel and environmentalmanagement lIould also increase and suchnovel stmtegies of coming up "ithcustomized bugs is an inevitable solution.but ethical issues surround the question ofmaking arti ficial organisms in the presenceof seemingly countless undiscoveredresources and the possibility of these bugsgetting out of control and causingunprecedented harm to the environment andultimately to us remains there.

The game is on for making virus-poweredbatteries and bio-solar cells with much moreefficiency than the inorganic ones.Astounding advancements in the field ofpharmacogenomics arc already bent uponrendering the 'old "ise' doctors obsoletefrom the field of medicine. They saycomputers will be able to diagnose theproblem/disease and counter it there andthen with customized. personalizedmedicines. Nanotechnology. through timelydiagnostics and targeted drug deliverysystems. is bringing the light against cancermuch closer to victory. Insulin lias one ofthe early fruits of recombinant DNAtechnology that saved millions of lives andg.enL'" therapy is bL'ing done these days toreplace dysfunctional genes with healthyones and it holds the promise of savingmany more in ncar IlJture. Today. Scientists

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who faced ethical dilemmas withtransplanting dead human organs resort tousing stem cells to make organs and thev trv-- ••.. ~ .to obtain the stem cells not by killing humanembryos but by reverse engineering of fatcells!

Life sciences provide a fertile field for theintersection of several scientific disciplinesto tap into the synergy of minds fromBiology, Chemistry, Physics. computerScience and engineering along with manyother fields, and Ihis science is likely to havea great impact ')n the society and economyon a global scnle in the near future. As thesocieties be':ome more' and morematerialistic, lone self-interest and desire todominate rais,:s heads and ills like terrorismemerge. The possibility of bio-terrorism; theinitial episo(.';s of which were seen asAnthrax envelDpes is very much real and itsconsequences can be far more disastrousthan those of any present day arsenal.Research shou:d go on to explore thewonders of natures that lie in the realm oflife sciences but governments and regulatoryinstitutions should be equally quick in takingappropriate action against any researchesthat are inherently sinister and counter-productive.

Meanwhile. more Brunos might not have toget burnt alier revealing their ideas due tothe far-reaching acceptance of Science andits fruits in this day and age. some firewould be needed to carryon blazing hotresearch. but some fire-extinguishers wouldalso be required to cool down the Brunoswho believe in vengeance.

Green Plastic-------A New Hope

Shenl'ee=a Slwukal

Dangling from electrical wires. hoveringhere and there. choking up drains and

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Biotechnology

messing up the streets. this is not a linetaken from a horror story, rather a scenefrom our own plastic driven, technologyfilled lives. A spectacle. whether going tothe school. office or university. This is thewonder of the 20"' century_' the age ofPLASTIC.

Everything seems to be \\ rapped up inplastic and becomes so easily incorporatedinto our lives because it is easy to carry.light weight. convenient and available in allsizes and shapes. One of the mostfundamental uses of plastics in our dailylives is in making plastic bags andcontainers used for carrying food items.groceries, fruits. vegetables etc.

Plastic bags and containers arc safe forkeeping cold items, but they arc not suitable(or preserving hot items because under hotconditions, plastics releases a highly toxiccompound called Bisphenoj that disrupts thehormonal system. It is a common practice toburn plastic in an open environment whichallows poisonous gases to be released intothe atmosphere resulting 111 vanousbreathing problems.

Despite all the benefits we get from them,there is a heavy price to pay. Plastics arcnon-degradable. This inertness comes fromthe fact that the carbon molecules arc sotightly trapped inside the. plastic lattice thatthis it allows them to remain unchanged loryears.

Plastic has more disadvantages than it hasadvantages. I must say that co,wentionalPLASTIC is 1'\0'1' FANTASTIC.

The major focus in this revolutionary fieldof plastic nowadays is GREEN PLASTIC.making life safe. convenient and eco-friendly. Scientists are trying to developgreen plastic. It docs not mean that it is

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The Scientific Ravi 20 I I

green In color. Technically. it IS

manufactured from plant sources (starch.cellulose). a renewable source rather than anon-renewable. It will reduce the risk ofdepletion of non-renewable resources.

PLA (polylactic acid) is a green plastic. It ismade from starch that is a natural polymerand is produced by plants duringphotosynthesis. Microorganisms act uponstarch that is obtained from wheat. corn etc.and transform it into lactic acid. Uponchemical treatment. it causes the molecule tobe lined up to form a long chain resulting inthe formation of a plastic called polylacticacid. One important property of starch is thatit is water-soluble. so a plastic made fromstarch may swell on hydration. limiting itsuse. "LA is very expensive due to which ithas not gained much attention.

It may be possible that genetically modifiedplastic can be formed from GM crops tooptimize plastic efficiency.

Naturally. bacteria arc the smallest plasticmanufacturing factory. Bacteria ean producegranules of plastic calledpolyhydroxybutyrate (PIIB) orpolyhydroxyalkonate (PHA). Genes frombacteria can also be cloned into corn plantswhich then manufacture plastic that can beobtained or modi lied chemically.

Great potential has been seen in this newinnovation of green plastic since it isdegradable and having been made fromplant sources. bacteria may be able todevelop the ability to degrade plastic.

But the term degradation is misunderstood.Degradation does not mean that the plastic isleli anywhere to degrade on its own: ratherthe degradation of plastic requires completecomposting' condition that will allow

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bacteria to act upon it under suitableconditions.

But the development of green plastic hasfueled a new debate that since it is madefrom plant sources. it will result indeforestation. Other points of concern havebeen raised such as If bacteria will developthe ability to degrade plastic. then all theplastic items in our homes would be eatenup. or whether green plastic is completelydegradable or not? If it is not completelydegraded then will the smaller particles leftbehind be hazardous for soil fertility or not')These are few questions that still need to beanswered and further work needs to be doneto make the green plastic practicable andaffordable.

The Promises fromBiotechnology to

Food Security in Pakistan

TaJ:l'aba Lali

Imagine the sweet. delicious mouth-watering brownies. desserts. cakes andmuffins on our table sans sugar. flour. cerealSlUffing and peanut oil ... surely horrible'Pardon iI' you su ITer Irom diabetes or arc ona diet. What else can a .lewd nigh/mare bethan the lack of this very basic necessity oflife" Pakistan. an agrarian country. isunfi.)rtunmely quite notoriously sufferingfrom a food shortage since the past decadeand this is hard to deem as agriculture is theeconomic mainstay of this country. Fertilesoils. laborious farming and high raw yieldsarc luminary in our agriculture buthoardings. losses due to natural calamities.low quality food and despised exportscounter the load sufficiency in our country.

UN statcmcnt on lood sccurity in Pakistanconcentrated on the competence of food.

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liberal access to it and reliability of itsprovision in the near future. The question is;will the Pakistani indigenous agriculturalsystem continue to provide ample foodsupply to meet this alarming demand andsubdue the shortage') The answer is no asmanifested: even the Green Revolution of1960s could not rescue us from today'sdismal shortage of sugar. wheat. cereal andother staple food crops. Experts havewarned that there is a threat of an expansivefamine and hunger in the country and it iscritical to be noticed as national sovereigntyis linked to food security.

The importance of biotechnology appliedonto our green fields was realized inPakistan back in 1960 when geneticengineering was emerging as a promisingscience of mounting lofty yield seeds andefficiently growing plants in some Far Eastcountries. The government of Pakistan hadestablished National Commission onBiotechnology in 2001 with the hope ofapplication of biotechnology specifically onagriculture sector. This biotechnology-fuels-agriculture strategy involved incorporationof crop-specific research, productivity andprofitability of farming. livestock.aquaculture production. agro-forestry, foodand cash crop. integrated pest managementand consideration of post-harvest losses.Agricultural biotechnology can thuspositively lead to beller and more food.

Success has, nevertheless. been reported inour country when it comes to pest resistantmaize. salt-tolerant tuber and rice crops,

'disease-free banana plants. and enormousyield per hectare of orchids. but these needexcessive advertisement for their activemass cultivation. Pakistan can most likelybenefit from the experience of China andIndia where the agricultural system andconditions are almost alike. The governmentneeds shiti the biotechnology frontage as a


Biotechnology

;'Vational Stratel-,.'V into high gear and learnfrom biotechnology practices employed inChina and India. India and USA aloneearned above 59.80 billion and $44 billionrevenue. respectively. by producinggenetically modi tied crops from year 1996to 2007. It should however be made clearthat biotechnology does not mean only usingGMO amended food. it also verses fornature's own means to enhance the varietyof. food crops. and to augment their netproduction.

With all its pros and cons. biotechnology'scapacity to eradicate food scarcity throughproduction of crops resistant to pests anddiseases. tolerant to adverse weather and soilconditions, crops that give higher yields perunit time. longer shell~lives, and possessingsuperior textures and flavors, cannot beunder estimated. We have a darker picture offood scarcity in the near future at one side.and a hopeful image of agriculturalimprovements by the pivotal aid ofbiotechnology on the other. To deal with thiscolossal issue, we need a compound policyconstituting our preferences and prioritiesfor Research and Development. investmentin food industry and processing sites. public-private sector affiliation. biosafety.intellectual property management and bypromotion of public awareness. Clicking onany such technology should be done underpolitical. ethical. economical and foreign-affairs bases since Pakistan is a signatory tothe Convention on Biological Diversity.World Trade Organization and CartagenaProtocol of Biosafety. In the foreseeablefuture there is a distinct possibility that ourefforts may be rewarded and we becomese1f~sufficient enough to proudly callourselves an agriculturally developedeconomy. Ilistory reminds us that it wasRussia whose food insecurity shook itsbackbone. For the sake of our poor. hungryand marginalized people. and for our


prosperous future. we must not evadebiotechnological applications confrontingthe food scarcity.

Mere Telomeres

AII/llil Ija:

Aging is a biological process that intriguesman throughout his life. It affects most cells.organisms and species while some unusualones appear not to be rusted by time at all.Human aging is accompanied bv anincreasing susceptibility to chronic di;easeslike Non-Insulin Dependent DiabetesMellitus. cardiovascular diseases and theonset of many genetic neurological diseases.Chalking out the factors affecting agingremains a myth but studies regardingtelomeres i.e.. telomere biology hasquenched this thirst somehow.

Elizabeth H. Blackburn. Carol W. Greiderand Jack W. Szostak were the pioneerinquisitors in this field. Telomeres arenucleoprotein complexes present at theterminals of chromosomes as protectivecaps. and playa crucial role in the integrityof genome. They consist of non-codin~double stranded nucleotide repeats andtelomere-specific associated proteins calledShelterin Complex. In humans. they are onaverage 9-15 kb long with guanineoverhang. that is. 5-300 bp long. Thisoverhang often coils back into the maintelomere helix and forms a T-shaped loop.This loop aids in mediating the formation ofend<ap. -

Each time the cell divides the genomereplicates with it. DNA polymerase-carriesout this replication in assistance with otherenzymes i.e., helicase. lipase. etc. DNApolymerase is never able to completereplication till the very 3' tip of the strandbeing replicated due to some 'natural flaws'

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in its mechanism so after everv round ofreplication about 30-200 bp are I~st from the3' terminal. If telomeres had not been therethis loss would occur directlv in the codin". ~region causing apoptosis (programmed celldeath) immediately.

After everv cell-division telomereshortening occurs until a critical length isreached and further shortening results incell-cycle arrest or cell-senescence.Approximately 50 doublings are enough tobring a cell to its crucial limit whichcompromises cell-viability. The telomerasc:a reverse transcriptase helps these telomeresto grow back hence compensating fortelomere loss in all those cells where it isexpressed. As an organism ages. the activitvof telomerase deteriorates and is not enougilto counteract the continuously degradi~gtelomeres. In humans this enzyme istypically found in cells with high 'divisionrates like germ cells and progenitor cells andduring embryonic development. Over-expression of this enzvme in a cell mav leadto tumor formation. - .

Telomere length is not affected bv onlv oneaspect mentioned above. but n;any 'otherparameters play an important role in thisphenomenon as well. These includeenvironmental factors: Reactive OxygenSpecies. psychological stress. adverse lifeevents. chronic and serious illnesses.malnutrition and childhood mistreatment.etc. The onset of aging is not only aided bvthe shortening of telomeres but also becaus~of lower telomerase activitv and increasedoxidative stress. .

When an individual consistentlv faces harshconditions. telomere attrition' fastens andeventually they are no longer able to attractthe Shelterin proteins or to form theprotective T-Ioops. On the other hand. inother circumstances the Shelterin proteins

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may themselves degrade affecting thegenomic safety. In either case cell death isinevitable. Experiments have shownphysical activity and exercise to bepositively associated with anti-agingbenefits like better telomere lengths inindividuals while smoking andsocioeconomic stress are in connection withshortened telomeres.

Telomere length and telomerase dysfunctionhave been shown to be significantlyassociated with age-related diseases likehypertension, atherosclerosis, heart failure.diabetes, etc. However, it should be kept inmind that this is not the only causative agentin these illnesses. Endothelial ProgenitorCells from hypertensive patients exhibit areduced telomerase activity whichaccelerates cell senescence. Also. shorttelomere length in these patients. especiallyin men, is likely to result in a higher pulserate. which is a commonly used clinicalmarker for artery stiffness indicatingcardiovascular vulnerability. In 200 I.Samani el al. conducted a populationresearch regarding the connection betweentelomere length and atherosclerosis. Theyproved that telomere length in leucocytes(white blood cells) was 303 bp shorter inindividuals with severe coronary arterydisease than in normal persons of the sameage. This made the patients approximately 9years older in health hence proving thatdevelopment of atherosclerosis is highlyassociated with cell-senescence induced bytelomere shortening. Chronic Heart failure(CHI') or simply heart attack is one of the'gifts' of advanced age. though it has beenseen to occur greatly in youngsters as well.has also been characterized by increasedmyocyte apoptosis and shortened telomeres.When patients with CHI' and normalpersons of the same age were compared. adifference of 40% was seen between theirtelomere lengths. Greater the loss. greater

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would be the severity of the disease.

Alzheimer's and Parkinson's disease areneurodegenerative ai Iments apparentlyassociated with aging. Alzheimer's patientshave shorter telomeres in their leucocytesand T-cells. Women suffer from this loss oftelomere length in T -cells more than mendo. This telomere story has been convolutedby research which demonstrates longertelomeres in the hippocampus of ADpatients. No information as to why thishappens has been put forward and the arearemains vague. It was further hypothesizedthat men who have short telomeres have alower risk of developing Parkinson'sdisease. This is highly contradictory to theprevious observations seen throughout age-related studies.

As individuals age, they experience adecline in body systems. Life span and howit ends is determined by many factors thatinteract in a very complex manner. Thecorrelation of telomere biology. aging andage-related diseases has been established bymany researchers. Variation in theexpression of telomerase gene is associatedwith better maintenance of telomere lengthwhich in turn ensures a protection from age-related diseases. better mental activity andlipid profiles leading the individual tohealthy and happy aging with longerexpected life-span. Future research focusingon life style and how it affects telomeraseactivity can be used to generate medicationand therapeutics that will strongly combatage-related diseases.

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Knockout Technology- AValuable Tool for Studying

Gene Function

Busil Jabbar

Studying the function of mammalian genesbefore the recent furtherance in appliedmolecular genetics was practically doneusing mutants generated by spontaneousmutations: however. this was problematiclargely due to the fact that other genesflanking the gene locus of the mutant genewere carried along with the gene understudy,

A breakthrough technology called"Transgenics" was developed later which.considering the mouse models. is theintroduction of a transgene (a foreign gene)in both the reproductive cells and somaticcells of a mouse using pronuclearmicroinjection, In pronuclearmicroinjection. the nuclei of the fertilizedcells are injected with the gene to be inserted(tr3nsgene) and the altered fertilized cellsare then implanted into female mice. Amajor drawback of this technique was thatresearchers could neither predict nor controlwhere in the genome the foreign geneticmaterial would be inserted resulting inrandom integration of gene into genome.Owing to that. the expression pattern wasinconsistent and the mice carrying the sametransgene displayed significantly varyingphenotypes.

To resolve the conflicts associated withtransgenics. scientists originated aninnovative solution called a genetic"knockout", By knockout technology. it ispossible to insert a defective gene at aprecise location in the genome of mice andhenceforward. to knockout or replace anormal or healthy gene with mutated or


Biotechnology

inactive gene. So. in knockout mice. one ormore genes have been disrupted or turnedotT by targeted mutation,

The reason behind the preference of mice inknockout experiments is that mice offercertain advantages for studying the biologyand diseases of humans because:

I. Mice and humans. being mammals.have much in common: theirdevelopment. behavior. physiology.body plan and diseases,

2. About 99% (almost all) mice geneshave their homologues present inhuman genome.

3. The genome of mice allows targetedmutagenesis in particular genes inembryonic stem (ES) cells. byhomologous recombination thusenabling efficient and precisealteration of genes,

There may be several differences 111procedures of knockout mice generation: atypical procedure involves:

I. Construction of a vector conslstll1gof mutated or altered gene andregions of DNA. which archomologous to the portions of targetDNA between which the originalgene is to be replaced with thealtered gene, One or more antibioticresistant marker genes are alsoincluded in the vector,

2. Insertion of vector DNA into theDJ\:A of embryonic stem (ES) cellsof mice by electroporation (usingelectric shock to insert DNA),

3. Selection of altered ES cells usingantibiotics and insertion of alteredcells into mice embryo.

-l. As a result. chimeric mice areproduced having some parts of theirbodies originating from the original

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5.

£OS cells and other parts from thealtered £OScells.The chimeric mice are crossed withthe normal mice to produce knockoutmice.

Fermented Foods

Bilqees Fa/ill/a/'h. D Scholar

Assigning a particular function to a gene isreallv challenging and requires significanteffo;t and vet knockout technology enablesus to ulili~e knockout mice as models inresearch to studY the function or effect ofthat gene ill l'il:o (inside the body) by itsreplacement with defective gene.

Knock-out mice acts as one of the mostadvantageous scienti tic tools to gain insightsinto th; human genome and its roles indisease. Utilizing knockout mice models tostudY genetic diseases at the molecular levelpro\;id~s better understanding of how aparticular disease is developed due to acertain gene. For example. knockout ofTP53 ge~e (which encodes for p53. a tumorsuppre~sor protein) has been extensivelyused in studying tumorigenesis (formationof tumors).

Hence. knockout technology acts as avaluable tool and holds immense potentialfor genetic and biomedical research.


Introduction

Agricultural crops are processed for manydi fferent reasons. These range from theremoval of anti-nutritional components andincreasing the storage lite of the finalproduct to adding value to increase bothemployment and income generatingopportunities. Fermentation is one of themost ancient and most important loodprocessing technologies. There istremendous scope and potential for the useof micro-on!anisms towards meeting thegrowing wo;ld demand for food. throughefticient utilization of available natural loodand feed stocks and the transformation ofwaste materials. Several individuals andorganizations are actively involved inresearch in this area.

Fermented toods 'are popular throughout theworld. They make a significant contributionto the diet of millions of individuals but areconsidered as the poor man's food. As soonas a familv can afford. they move away fromcarrying ~)ut home teflll~ntation. This is apity because fermented food products havemany nutritional advantages \\ hich surpassthe ;vestern-style fast foods and processedfoods.

It is often felt that traditional products madeat the small scale are unhygienic and unsafe.This is sometimes true. However the case isoften overstated. Many lermented foods arcinherentlv sate due to low moisture contentsor high ~ciditv. Quality control proceduresare ;ssential' for the production of saleproducts and contribute to the success ofsmall lood processing businesses. Theseprocedures need to be developed with the

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processors who must understand and applythem.

However. scientists and policy makers haveneglected this area. particularly traditionalfermcnted products from developingcountries.

Fcrmented Foods: an ancicnt tradition

The lirst fermented foods consumcd wereprobably fermented fruits. Therc is reliableinformation that fermented drinks werebeing produced about 7.000 years ago inBabylon (now Iraq). 5.000 years ago inEgypt. 4.000 years ago in Mexico and 3.500years ago in Sudan. Bread-making probablyoriginated in Egypt over 3.500 years ago.Fermentation of milk started in many placeswith evidence of fermented products in usein Babylon over 5.000 years ago. There isalso evidence of fermented meat productsbeing produced for King Nebuchadnezzar ofBabylon. China is thought to be the birth-place of fermented vegetables and the use ofAspergillus and Rhi=vpus moulds to makefood. The book called "Shu-Ching" writtenin the Chou dynasty in China (1121-256Be) refers to the use of "chu". a fermentedgrain product.

The Need for Rescarch

Although fermentation of foods has been inuse for thousands of years. it is likely thatthe microbial and enzymatic processesresponsible for the transformations werelargely unknown. It is only recently thatthere has been a development in theunderstanding of these processes and theiradaptation for commercialization. There isgreat need for research in this area toachieve new goals.


Biotechnology

Objcetivcs

This article presents an overview of thelennented fruit and vegetable products ofA Ii-ica. Asia and Latin America. The aims isto introduce the reader to the vast wealth ofknowledge. much of which is indigenousand undocumented and the importanceattached to fermented fruits and vegetablesin the diet.

Bencfits of Fermcntcd Foods:

Fcrmentcd Foods are Culturally andEconomically Important: Fermentation is arelatively efficient. low energy preservationprocess which increases the shelf lile anddecreases the need for refrigeration or otherform of lood preservation technology. It istherefore a highly appropriate technique lorusc in developing countries and remote areaswhere access to sophisticated equipment islimited.

Bones and Hides:

A . wide range of "waste" products arefermented to produce edible food productsin Sudan. These include bones. hides andlocusts. Fresh bones are fermented into avariety of products. "Dodery" is producedby chopping bones into small pieces andplacing them into fermenting vats. Anotherproduct "Kaidu dig/a" is made from thevertebrae of the backbone.

Use of Waste Products in Indonesia:

In Indonesia. a variety of waste products isfermented to produce nutritious foodproducts. Telllpe-hongrek is a protein richfood made in Indonesia by fermentingpeanut and coconut press-cake, remainingafter oil extraction. The product is similar totraditional tempeh produced from thefermentation of soya beans. The production

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The Scientific Ravi 2011 Biotechnology

Some Fermented Food Products

3, Banana Beer:Throughout. Alrica Banana beer is madefrom bananas. mixed with cereal flour.Un\'{/ga banana beer in Kenya is made frombananas and sorghum or millet and Lllhisi ismade from bananas and sorghum.

.t. Cashew \Vine:Cashew wine is made in many countries inAsia and Latin America. Cashew wine is alight yellow alcoholic drink prepared fromthe fruit of the cashew tree (A/Icardillllloccidentale). It contains an alcohol conlentof between 6.0-12 %.

L Red Grape Wine:Red grape wines are made in many African.Asian and Latin American countriesincluding Algeria, Morocco and SouthAfrica. It is made from the fruit of the grapeplant (Vitis I'i/lijera). There are widevarieties of grape used including CabernetSauvignon, Grenache. Nebbiolo, Pinot Noir.and Torrontes.

day 90-95 % of thewill have been

2. White Grape \Vine:White grape wines are made In manyAfrican, Asian and Latin Americancountries including Algeria, Morocco andSouth Africa. It is prepared from the fruit ofthe grape plant (Vitis vinijera). There aremany varieties used including Airen.Chardonnay. Palomino. Sauvignon Blancand Ugni Blanc. The main differencebetween red and white wines is the earlyremoval of grape skins in white wineproduction.

over and by the thirddangerous chemicalhydrolyzed.

Removing Cyanide by Fermentation:Cassava contains a naturally occurringchemical: cyanogenic glucoside. Wheneaten raw or improperly processed. thissubstance releases cyanide into the body,which can be fatal. Correct processingremoves this chemical. The cassava is tirstpeeled (as about 60-70 % of the poison is inthe peel) and then soaked in stagnant wateror fermented in sacks for about three days. Itis sometimes grated or rasped as this helpsto speed up the fermentation process. At thebeginning of the fermentation. Geotriculllcandida acts onthe cassava. This tends tomake the product acidic, which tinally killsoff the micro-organisms as they cannot existin such a medium. A second strain of micro-organisms (Cornibacteriulll lactii), whichcan tolerate the acidic environment then take

of Tempeh- bongrek is mold fermentation.initiated by inoculation of the soaked.acidi tied press-cake with Rhizopus species.The inoculated cakes are placed on bananaleaves or plastic sheets in a dark room forabout 2 days. An incubation temperature of37"C is optimal for the mould and preventsthe growth of P. cocovenenans. whichproduces Bongkrek toxin. A pH of less than6.0 also prevents the development ofBongkrek toxin. Ontjolll is produced fromwaste groundnut press cake, tapioca wasteand the solid waste of Tahu.lt is preparedusing a mixed culture of micro-organismswith Rhizopu.l' or Neurospora speciespredominating and is mainly produced inwest Java where it is consumed as a sidedish in the form of deep fried slices. It formsan important daily food item for the westJavanese, particularly those from the lowerincome groups. Fresh coconut residue. leftover from the production of coconut creamor milk, can be fermented by Bacillussubtillis, in an alkaline fermentation. toproduce selllayi. which is widely consumedin Indonesia.

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5. Tepaclle:Tepuche is a light. refreshing beverageprepared and consumed throughout Mexico.In the past, tepache was prepared frommaize. but nowadays various fruits such aspineapple, apple and orange are used. II'fermentation is allowed to proceed longer. itturns into an alcoholic beverage and laterinto vinegar. The microorganisms associatedwith the product include Bacillus subti/is, B.graveo/us and the yeasts. Tal'll/apsisinsconspicna, Saccharomyces cerevisiae andCandidaqueretana.

6. C%llclle:C%nche is a sweet. fizzy beverageproduced in Mexico by fermenting the juiceof the fruits of the prickly pear cacti -mainly Opuntia species. Tibicvs aregelatinous masses of yeasts and bacteria.grown in water with brown sugar. They arealso used in the preparation of tepuche.

7. Fortified Grape Wines:FOl1ified wines are made in the Republic ofSouth Africa and NOl1h Africa. FOl1ifiedwines are made by adding spirits to wines.either during or after fermentation. with theresult that the alcohol content of the wines israised to around 20 %. i.e. approximatelydouble that of table wines.

8. Date Wine:Date wines arc popular in Sudan and Nol1hAfrica. They are made using a variety ofmethodologies. Daklwi is produced fromdates. E/ mad.fima is produced by buryingthe eal1henware pots underground. Bentimerse is produced from a mixture ofsorghum and dates. Nebit is produced fromdate syrup.

9. Sparkling Grape Wine:Sparkling grape wines are made in theRepublic of South Africa. Sparkling winescan be made in one of three ways. This

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involves adding a special strain of wineyeast (S. cerevisiae var. e//ipsoideus) - achampagne yeast - to wine that has beenal1ilicially sweetened.

10. Jack-fruit Wine:Jack-fruit wine is an alcoholic beveragemade by ethnic groups in the eastern hillyareas of India. As its name suggests. it isproduced from the pulp of jack-fruit(Artocarpus heterophyl/us). A little water isadded to the pots along with fermented wineinoculum from a previous fermentation. Thepots are covered with banana leaves andallowed to ferment at 18-30°C for about oneweek.

II. Fermented Plant Saps:Vil1ually any sugary plant sap can beprocessed into an alcoholic beverage. Manyalcoholic drinks are made from the juices ofplants including coconut palm, oil palm.wild date palm. nipa palm, raphia palm andkithul palm.

12. Toddy:Throughout Asia. pal1icularly India and SriLanka, Toddy is an alcoholic drink made bythe fermentation of the sap from a coconutpalm. It is between 4.0-6.0 % alcohol andhas a shelf life of about 24 hours.

13. Pulque:Pulque is the national drink in Mexico.where. it is claimed. it originated with theearly Aztecs. Pulque plays an impol1ant rolein the nutrition of low income people inMexico with B vitamins being present innutritionally impol1ant levels. It is obtainedby fermentation of aguamiel. which is thename given to the juices of various cacti,notably Agave alrol'irens and A. Americana.

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1.4. U/allzi (Bamboo Wine):In East and Southern Africa. Ulanzi is afermented bamboo sap obtained by tappingyoung bamboo shoots during the rainyseason. It is a clear. whitish drink with asweet and alcohol ic flavor.

15. Basi (Sugar cane wine):Basi is a sugar cane wine made in thePhilippines by fermenting boiled. freshlyextracted. sugar cane juice. A similarproduct called sholo sake is made in Japan.

16. IIJuralilla:,Humlina is an alcoholic drink made fromsugar cane and mumlina fruit in Kenya.

17. Dry Salted Lime Pickle:Dry salted lime pickles are produced in Asiaand Africa. They are particularly popular inIndia, Pakistan and North Africa.

18. Pickled Cucumbers:Pickled cucumbers are made in Africa. Asiaand Latin America. Cucumbers undergotypical lactic acid fermentation. Khalpi is acucumber pickle popular during the summermonths in Nepal.

19. Pak-Gard-Dong (Pickled LeafyVegetable):Pak-Gard-Dong is a fermented mustard leaf(Brassica juncea) product made in Thailand.Micro-organisms associated with thefermentation include Laclobacillus brevis.Pediococcus cerevisiae and Laclobacillusplanlarum.A similar product (Hum choy) ismade in the South of China. This isproduced by fermenting a' local leafyvegetable.

20. Tempoyak (picklcd Durian):Tempoyak is the fermented pulp of a durianfruit (Durio zibelhinus) from Malaysia.

GC Univcrsity, Lahorc

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21. Pickled Beetroots:In Russia beetroot is pickled by cleaning.slicing and placing in a container with salt.

Conclusion:

Fermented foods should be recognized as apart of each countries heritage and culture.and efforts made to preserve the methods ofproduction. A recognized body (governmentor non-government) should take theresponsibility for the collection of detailsand the promotion of fermented foodproducts. Consumers need to be made awareof the numerous benefits of fermented foodsand their prejudices against fermented foods.especially those traditionally produced at thehome scale, dispelled.

Interesting Applications of Hair

Kanwal JVazir

What notion comes to your mind when youthink about your hair? I guess it varies fromperson to person. Some people may thinkabout the way they maintain and style theirhair. others may admire the natural beauty oftheir hair and still for some others. thisnotion may even bring grimace to their facesas according to them they have to carry theburden of having a pile of hair on theirheads. But what about the scientific mind?There are such dexterous minds on this earththat can jolt your brain even in this respect.Most of us have already apprehended themanipulation of hair for DNA fingerprintingtechniques. But here I would like to bringforth two novel uses of hair.

When talking about simple chemistry of hair.these are thin compiles of keratin and deadcells. This very 'simple' chemistry has blessedour hair with anomalous traits. One of which isthat our hair can absorb oil from water and canact as natural sponge. This phenomenon IS

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being brought into use to abate the dilemma ofoil spills. The idea evolved in u.s. by ahairdresser Phil McCrory who got it patent in1995. In 1989. Alaska suffered an oil spill.McCrory was watching its coverage ontelevision and because of his keen observation:he noticed that an otter's fur was clamped withoil as it had absorbed the oil from water. Thisobservation inspired him to test if hair could beused to clean the water from oil. So he came tothe conclusion that oil actually clings to thehair. Various studies and tests have followed it.

As oil spi lis threaten our environment withmany hazards. Aquatic liIC is at great riskwhere oil is spilled. The health safety ofhumans is equally at stake due to its link withaquatic life. So oil spills need to be counteredas soon as possible. Iluman hair can becollected from differem saloons and convertcdinto a compact mat. Fungi like oystermushrooms are also added to these mats so thatatier the take up of oil. it can be degraded intoless toxic or non-toxic compounds. Animal furand feathers can also be used. However, somescientists are skeptical about this technique.They say that handling is difficult and theworkers show hesitation. \loreover. thedegradalion rate of hair is slow and fur andfeathers may exude unpleasant odor with thepassage of time. implying that we could end upwith an even bigger problem than we startedwith. More research is needed in this aspect.

A noteworthy example of Manila. the capital ofPhilippines. should be quoted here. The peopleof Manila initiated a campaign called "Stop theOil Spill" to combat the oil spill of 2006.Everyone took part very enthusiastically. ;'vlanyprisoners shaved their heads and chests toeontrjbute 10 the campaign. Hair saloons alsohelped. Many volunteers came forward todonate their hair.

Let's come to the other amazing use of hair.What do you usually do if a hair accompanies


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your food in its journey to your mouth? Do youleel nauseated? Then how would you feel uponbeing told that hair can be used in makingfood? Many of us add soy sauce in fOLlli itemsat our homes or we consume it in the form ofother market foods and I()od products. llair canprovide a rich alternate to soy sauce. Humanhair proteins are as nutritious as the originalingrcdients of soy sauce i.e. soy beans. andwheat bran etc. These proteins can be extractedand used for soy sauce production. Bakers arevery attracted towards th is prospect due to thepresence of cysteine that gives specillc textureto baking and improves it.

This has been illegally practiced in a Chinesecompany ..If()l1gshllo; Soy SOllce ... Peoplewere unknowingly consuming it atcomparatively lowcr ratcs. A fter inspection itwas reported that the lilCtory was able toextract 100.000 tons of amino acids from hairdaily which were then converted into powderor syrup form to make it easy to handle.Alterwards. it was diluted up to -10 times andthen bottled into soy-sauce-labeled bottles.Thcy called this. "bioengineering".

Many harmful non-food additives like solidhydroxide and hydrochloric acid are also addedto it. In their view it is done to make itconsumable by humans. On investigation. itwas disclosed that they were nO! ready toconsume it themselves. The irony in this is bestsummarized by the following quote:

"One person's recycling is another person'sretching ov.:r a toilet"

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Liquid FermentationBasil Jabbar

Considering the biochemical View offermentation. it is defined as onlv ananaerobic process that occurs in the livingcells when the oxygen supply is low.However. industrial biotechnologv andmicrobiology which involve ;:;equentplaying with the microorganims haveoriginated a much broader definition of thefermentation process and describe it asbeing an aerobic or anaerobic mass cultureof microbes for getting the dcsired product.This product can bc an enzyme. ametabolite. a recombinant product. a bio-transformcd product or microorganismsthemselves (microbial biomass e.g. SingleCell Protcin or SCP). ~ ~

Fermcntation- although the process got itsname many centuries later. it's history canbe traced as far back as thc ancient timeswhere the process of wine making wascarried out in pots by people using fruitjuices containing microbes. Fermel1lationwas initially utilized mostly for theproduction of food products but as scientificadvances were made. it expanded itsbranches and it was made applicable for theproduction of products like enzvmes.vaccines. antibiotics etc. These producis cannow be produced in the industrics in largetanks called bioreactors or fermente~sspeci fically called liquid cu Iture fermenters.This type of fermentation process is referrcdto as Submerged or Liquid Fermentation(shonly stated as SmF) wherebymicroorganisms are mass cultured in aliquid medium for the production of thedcsired product.

Large excess of solvent (water) needs to beused while formulating the medium tomaintain submerged conditions for mediumcontents. Hence. in addition to supplying all


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the nutrients essel1lial for microbial gro\qh.water is sufficiently added and the ;atio ofother medium contents to water may be upto I :25. Steps of Sm F process stan frominoculum preparation (wherebv inoculumcontaining microorganisms to be inoculatedinto the fermentation medium is prepared)and go through media formulation.sterilization. fermentation at requiredconditions. product recovery and packaginguntil it finalizes at eftluel1l treatment.

Another fermentation technique. Solid-StateFermentation (in shon. SSF) in whichmicroorganisms are cultured in solidmedium is better in product yield andeconomically feasible for the production ofcenain products (sccondarv microbialmetabolitcs e.g. alkaloids).' Yet. SmFremains the widely practiced technique bythe fcrmentation industries of the worldtoday due to its wider applications and anadvantage of better control over factors liketemperature and pH which arc diflicult to beregulated in an SSF process. Overall. SmF isan easicr system to work with. The substratehandling in SmF is easier than SSF. It ismuch simpler and easier to agitate liquidsthan to move solids from one place toanother and to sterilize a large volume ofliquid than that of solids. ~

SmF has a broad range of applications inindustry for the production of enzymes.al1libiotics. vaccines. monoclonalantibodies. 1()Od products (yoghun. bread.cheese and other baked products) andbeverages. Recombinant products (e.g.hormones and interferons) by GlvlOs(Gcnetieally Modified Organisms) arc alsoadvantageously produced by SmF bvculturing GMOs. The yeast used in bakin~and bre~\ing (Baker's' yeast and Brewer'~yeast: .~'lIccharomyce:" n:re\'h;iae) is culturedby this process. Thanks to SmF that we are

lot

The Scientific Ravi 201 I

getting a lot of fermentation products that;re o(an appreciable significance and use.

Sa crificia 1-flesh -th rowin gMentality contributing to the

Deterioration ofEnvironmental Conditions of

Lahore

AIII/wmmad Saad Farooq

We arc Muslims and charity is one of theimportant teachings of our religion Islam.We. theretore. spend in the name of Allah inmanv ways. In our society sacrificial-flesh-thro~ving . is getting popularity each day.People at canal and river banks offer theblood-stained bags containing foul smellingflesh in them causing serious environmentalproblems. mostly overseen by us. furthercOlllributing to the deterioration ofenvironment of our city.

Pakistan has a very low literacy rate and thepeople arc victimized by the superstitionswhich have nothing to do with "Sadqa".There are five main types of charity whichinclude charitv on wealth. tongue. position.- -wisdom and knowledge.

Throwing these flesh bags causes a numberof problems such as:

I. Polyethylene bags are non-degradable. so microbes includingbacteria and fungi etc. cannotremove them th)m environment andthev remain. as thev are. tilr decades.

2. Th~se plastic bags' get stuck into thesewage systems and block the pipes.

J. Thev are also barriers in thereC\~c1ing processes such asCOI;lposting or biogas productionunits and need to be screened out

GC UniHrsily. Lahore

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causing an additional cost to theprocess .

.to ;\loreover. they also affect theaesthetic preference and beauty of acertain area.

5. Flesh sold in these blood stainedbags is sub-standard cow lungs\\I~ch is not even good for animals.

6. Blood is an excellent medium formicroorganisms. so it leads to thepropaga;ion of lethal and non-lethalmicrobes.

7. In nature crows and kites etc. arcprogrammed to remove the deadanimals as soon as possible from theenvironment and use them as theirfeed. but due to this practice they areattracted to some speci fic sites fromwhere they can get flesh effortlesslyin excess.

8. Th"ese animals arc vulnerable tohealth risks filr overeating with leastslnl~1.d~.

9. [)ue-~to deviation in behaviors ofthese animals and birds. deadanimafs arc subject to theenvironment where they smell fouland are prone to micro flora.

10. These dead animals can be excellentnourish ing niches for the pathogen icmicmbes to flourish and causeserIOUS health risks tor thecommunity.

II. These plastic bags also disturb theayuatic life in the rivers badly.

12. This single practice causes muchharm to the various organisms andconseyuently disturbs the foodchains and webs.

Islamic teachings. portraying the importanceof conservation in Islam. go on 10 say:

I. "There shall be no damage and noinllictioll of damage." (AI-Hakim)

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2. "And do not do mischief on theearth. alier it has been sci in order"'(Quran 7:56).

3. Hazrat Anas reported that theProphet (saw) said: "If a Muslimplants a tree or cultivates a crop. thenwhenever a bird or a man or ananimal eats from it. it will beconsidered as a charity for him. "(AI-Muslim)

From these Verses and Ahadees. we can saythat Islam is purely an environmemalfriendly religion and will never allow anything that harms the environment. So wecannot simply deny the importance of theenvironmental conservation because ourreligion also pays much stress upon if.;-"10reover throwing the sacrificial tlesh inthe plastic bags is playing a vital role in thedeterioration of the environment of Lahore.Crows and kites die in an ample number forengulfing plastic bags. Our sewage systemblocks creating health issues. the site alsoacts as one of the best suitable places for thepathogenic microorganisms to grow on theorganic mailer and they are carried by tliesand mosquitoes to the surroundings. Sothrowing foul-smelling tlesh in the wateryarea is not a preferable way to give charity.instead we must employ environmentalfriendly ways i.e. natural ways like plantinga tree . contributions in increasing the

fertility of soil etc.


Biotechnology

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