We all have been familiar with many infective diseases since many centuries ago. Some bacteriums have killed 1000000s of our lives, some viruses have great possible to devour many people wellness and wealth and both are still enormously stalking our world. We did detect Penicillin, scientists have developed many antibiotic and antiviral drugs to kill and battle against these bacteriums and viruses. This is the war that will hold no terminal.
I have been believing and analyzing about these micro-organisms since my undergraduate grade stressing about the diseases and their basic characteristics. In this survey, I wanted to research many facts about the bacteriums and viruses for advantages of our medical scientific disciplines and I found out that there had been many researches and finds about utilizing the bacterium and viruses for our goodness and surprisingly there will be more and many potency for our future medical scientific disciplines.
The most interesting thing I have studied is the procaryotic viruses called Bacteriophage and they truly have the really strong potency to be used as a arm against many infective diseases including multi drugs resistant bacteriums infection and against malignant neoplastic disease such as really lifelessly encephalon malignant neoplastic diseases and even the possible remedy of many types of malignant neoplastic disease by selectively aiming merely the malignant neoplastic disease cells without impacting the normal 1s and I besides have studied about the telomerase enzymes that have the possible against human cellular ripening.
Aim OF MY INDEPENDENCE STUDY
As we are populating on the universe interacting with the ecosystems incorporating different kinds of unicellular and multi cellular beings, most of our developments and pros and cons are tightly associated with these beings and the first aim of my survey is to cognize or associate the good effects we may obtain from our beings by understanding them and besides understanding ourselves scientifically.
To cognize and if possible, to suggest or to do the stairss to develop really effectual possible hereafter anti malignant neoplastic disease intervention utilizing bacteriophage.
To suggest the possible anti-aging mechanism utilizing bacteriphages.
To develop drug which can be efficaciously used for the many multi drug immune bacteriums infections such as multidrug immune Tuberculosis utilizing bacteriophages and to place the possible methods for the drug development and their several infective diseases with the delightful and cherished aid from my supervisor
After analyzing and stressing upon the general chief advantages that we get from the bacteriums and virus for our medical scientific disciplines, I want to concentrate my survey upon the Bacteriophage viruses which can perchance be used as a vector for cistron therapy and cistron ordinance for my desire against aging of human being and in another word against our inevitable portion of our human life called decease.
Another range is to utilize the phage as really specific malignant neoplastic disease cell killing agent for many tumours incorporating specific surface markers or receptors such as encephalon malignant neoplastic diseases.
I want to analyze by reading books and diaries and besides with my advanced intelligent measure by measure, from general to inside informations and to work out all the inquiries every bit much as I can and so to suggest the really new techniques utilizing molecular degrees and receptors degrees.
If we want to cognize something, we must foremost understand it fundamentally and so, my really foremost of import thing to make is to cognize about the general of import and some really detail features of the bacteriums and viruses. Without the general cognition of them, it is non possible to happen out more about them. Many mention books and internet links and assist me with this portion and to cognize the many good effects of them for Medical Sciences.
After this, my of import program is to analyze profoundly inside the bacteriophage viruses and their current and future use for Medicine and so accessing the cognition with my advanced thought and advices of my supervisor I hope that I will be able to larn, survey and happen many things about for Medical Sciences.
2.1: Introduction to bacteria
All procaryotic beings are classified as bacteriums and they are divided into eubacterium which includes all the bacterium of medical importance and archae-bacteria which is a aggregation of evolutionarily distinguishable beings.
Typical BACTERIA: Most of them have forms such as rod like, sphere or bottle screw. Their cells are smaller than the eucaryotic cells and all of them except the Mycoplasma have the stiff cell wall environing the cell membrane. Apart from the forms, the cell wall defines whether the bacteriums are Gram Negative or Gram Positive.
Degree centigrades: Documents and SettingsuDesktopmycoplasma_m.jpgFigure: Electron micrograph of MYCOPLASMA *which has no cell wall
Bacteria cells reproduce by binary fission. Atypical Bacterias are the distinguishable bacterium groups missing of important features structural constituents or metabolic capablenesss. They includes Chlamydia, Rickettsia and Mycoplasma etc.
2.2 ADVANTAGES OF BACTERIA FOR OUR BODY AND MEDICAL SCIENCE
Although bacteriums can do many diseases and wellness jobs to human being, they besides have many good effects for our human organic structure and medical scientific disciplines.
NORMAL FLORA: Many different microorganisms largely bacteriums are continuously populating the human organic structure without giving any injury. Human organic structure is normally unfertile when a healthy new born enters the universe. But, after birth, the organic structure acquires normal vegetations from the environment and nutrient. The really of import fact is that the species of that vegetations can non be stiffly defined because they differ from one person from another as a consequence of physiological differences, diet, age and geographic home ground.
NORMAL FLORA AGAINST THE INVADING HARMFUL INFECTIONS
The bacteriums need receptors and foods for their metamorphosis. The incursive infections will confront with the competency of normal vegetations for these indispensable receptors and foods. Some bacteriums of the intestine can even bring forth the antimicrobic substances so that the occupying beings can be killed. But, the substances bring forthing bacterium themselves are immune to their ain substances. These effects can cut down the possibility of the infective diseases and act surprisingly as a defence mechanism against the infections.
GERM FREE ANIMALS: The important of the normal flowered are now good explored by analyzing the germ free animate beings which have no normal vegetations as conventional animate beings. They are produced by particular cesarian subdivisions and so they are maintained in particular isolators. Experiments showed that in the germ-free animate beings, the alimental lamina propria is developing, the motility of the GI piece of land is reduced and the enteric epithelial reclamation rate is merely half of the normal conventional animate beings.
In surveies with antibiotic treated animate beings besides suggest that the normal vegetations can protect our organic structures from the pathogens. The research workers foremost treated the animate beings with Streptomycin to cut down the normal vegetations and so made them infected with the Streptomycin immune Salmonella bacterium. In normal status, approximately 10000000 Salmonella were needed to do the set up infection but in Streptomycin pre treated 1s, merely 10 beings were needed to do infection.
NORMAL FLORA FOR OUR IMMUNE SYSTEMS
Bacteria colonisation of a new born baby is the really powerful stimulation for the development of immune system. The surveies showed that the antibodies concentration after infections is significantly reduced in germ-free animate beings bespeaking the defect in acquired immune system.
Bacterias are besides the of import suppliers of of import foods such as Vitamin K and they besides help with digestion and soaking up of foods.
LIPPINCOTT ‘s ILLUSTRATED REVIEWS of MICROBIOLOGY 2007 EDITION INTRODUCTION TO BACTERIA, page 1 -6 Normal Flora Page, 7 – 10
KAPLAN USMLE TEXT BOOK ( MICROBIOLOGY ) 2009 Edition
HARRISON ‘S PRINCIPLES OF INTERNAL MEDICINE 2006
3.1 INTRODUCTION TO VIRUSES
A virus is an infective agent incorporating genome which is either RNA or DNA and a protein mirid bug designed to protect the genome. Many viruses have extra construction like envelope which is protein incorporating lipid bi-layer. The kind of nucleic acid in the virus is the most cardinal and of import of belongingss of virus. The nucleic acid may be individual stranded ( ss RNA US Secret Service DNA ) or dual stranded ( ds DNA or ds RNA ) .
The Single stranded RNA genome are subdivided into the positive ( + ) mutual opposition which is, of courier RNA sense that can be used as templet for protein synthesis. Negative ( – ) mutual opposition or antisense which is complementary to the messenger RNA sense and so they can non be used as temperate for protein synthesis straight.
Degree centigrades: Documents and SettingsuDesktop5.jpgFigure: Influenza viruses
3.2 THE REPLICATION CYCLES OF VIRUSES
Degree centigrades: Documents and SettingsuDesktopVirusenvelopedcycle.jpgFIGURE: THE LIFE CYCLE OF THE HIV ( ENVELOPED ) VIRUS
The rhythm begins with the fond regard of the virus to the host cell called surface assimilation stage,
The initial fond regard of a virus to the host cell is with the interaction between specific molecular construction on the surface of the virus and receptor molecules in the host membrane that can acknowledge the construction. The receptor molecules on the host cell membrane are specific for the household of the viruses and they are the molecular constructions that normally carry out normal cell maps. The receptors for the viruses are present merely on specific cells or are alone for one animate being species. So, the absence or presence of the host cell receptors is so of import finding for the susceptibleness or opposition of a species to a given virus. If we can genetically pull strings the specific receptor affinity for the viruses, we will be able to assail or kill or alter the desired targeted cells.
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Figure: HIV virus adhering to the cell, fond regard is accomplished by the SU fragment of the env cistron merchandise on the surface of the HIV which binds to the CD4 molecule. So, the HIV viruses can merely infect the assistant T cells, monocytes and dendritic cells which contain the CD4 protein in their cell membrane.
The two mechanisms the virons enter the cells traversing the cell membrane are the receptor-mediated endocytosis: the viron binds the cell surface receptor and the cell membrane invaginates enveloping the virion in and endocytotic cyst ( endosome ) . The virion so enters the cytol by assorted mechanisms depending upon the viruses. It is facilitated by one or more viral molecules in general. Degree centigrade: Documents and SettingsuDesktopendoem.jpg degree Fahrenheit
Membrane Fusion: Some enveloped viruses enter the cell by merger with their envelopes with the membranes of host cells. Glycoproteins of the envelope can advance this and viral membrane so still remains associated with the plasma membrane of the cell and merely the nucleocapsid is released into the cells. HIV viruses enter the cells by this manner.
Degree centigrades: Documents and SettingsuDesktopvirus001.jpgFigure: Left side shows the incursion of the virus by ENDOCYTOSIS while the right screening FUSION
This is the stepwise procedure of dismantling of the viron that enables the look of the viral cistrons that carry out viral reproductions. Most of the stairss occur inside the cells and depend on cellular enzymes and in rare occasions, freshly synthesized viral proteins are needed to finish the procedure. The loss of one or more structural constituents of the viron will take into the loss of ability to infect another cells reflecting as the eclipse period of the growing curve.
Degree centigrades: Documents and SettingsuDesktopssdna.gifFig: the reproduction rhythm of individual stranded Deoxyribonucleic acid virus
Deoxyribonucleic acid virus reproduction: There is a broad macromolecular event fluctuation between households of viruses for the reproduction processes depending chiefly upon the viral genome sizes. The smaller the viral genome, the more the virus must depend on the host cell to retroflex. Besides the mechanisms of reproductions for US Secret Service DNA viruses and Ds Deoxyribonucleic acid viruses are different.
Degree centigrades: Documents and SettingsuDesktopviral repli Ds dna.gif
Figure: the reproduction rhythm of dual stranded Deoxyribonucleic acid virus
RNA virus reproductions
Type 1: Ribonucleic acid viruses with a individual isolated genome of ( + ) mutual opposition that replicates with complementary ( – ) strand intermediate. In this, the infecting parental RNA serves as both messenger RNA and subsequently as a templet for synthesis of the complementary ( – ) strand.
Type 2: Viruss with ssRNA genome of ( – ) mutual opposition which replicate with a complementary ( + ) strand intermediate. ( – ) mutual opposition genomes have two maps, one is to supply information for protein synthesis and the 2nd is to function as templet for reproduction. But they can non carry through without anterior building of complementary ( + ) strand intermediate.
Type 3: Viruss with ds RNA genome: dsRNA genome is segmented, with each section coding for one polypeptide. But, the eucaryotic cells do non hold the enzyme to transcribe dsRNA. So, messenger RNA transcripts are produced by virus-coded, RNA dependant RNA polymerase ( RNA polymerase ) located in the sub viral nucleus atom. This atom contains dsRNA genome and associated viral protein, including the RNA polymerase. In reproductions, the ( + ) RNA transcripts are non merely used for interlingual rendition, but besides as templets for complementary ( – ) strand synthesis, ensuing in the formation of dsRNA offspring.
Type 4: Viruss with a genome of ssRNA of ( + ) mutual opposition that is replicated with with a Deoxyribonucleic acid intermediate: the transition of a ( + ) strand RNA to a double-stranded Deoxyribonucleic acid is accomplished by an RNA-dependent Deoxyribonucleic acid polymerase, normally known as “ rearward RNA polymerase ” , which is contained in the virion. The ensuing dsDNA becomes integrated into the cell genome by a viral integrase ‘s action. Viral messenger RNA abd offspring ( + ) strand RNA genomes are transcribed srom this integrated Deoxyribonucleic acid by the host cell RNA polymerase.
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4 ASSEMBLY AND RELEASE OF PROGENY VIRUSES:
Degree centigrades: Documents and SettingsuDesktophiv_bud.jpg Fig: negatron micrograph demoing budding and release of HIV1 atoms
The assembly of the nucleocapsids by and large occurs in the cytol for most RNA viruses and in the karyon for most Deoxyribonucleic acid viruses where the viral nucleic acerb reproductions take topographic point.
LIPPINCOTT ‘s ILLUSTRATED REVIEWS of MICROBIOLOGY 2007 EDITION, Unit-Viruses, 233-243
KAPLAN USMLE TEXT BOOK ( MICROBIOLOGY ) 2009 Edition
HARRISON ‘S PRINCIPLES OF INTERNAL MEDICINE 2006 EITION
Introduction to Bacteriphages
Bacteriophages are the viruses that replicate inside the bacterial cells. It contains nucleic acid encapsulated by the protective protein coat. The nucleic acid may be DNA or RNA depending on the phage and may be individual stranded and some are dual isolated ranging from the length of 3000 bases to 200,000 bases.
Degree centigrades: Documents and SettingsuDesktopacteriophage2.jpgThe electron micrograph demoing the construction and size of a Bacteriophage
The reproduction starts with the fond regard of the virus to the receptors of the cell surface of bacteriums. Then the phage injects the nucleic acid into the cell go forthing all or most of the protein outside the cell. This is the obvious difference between the virus that infects the craniates and the virus that infect the bacterium. In former instance, the virus is wholly taken up by the cell and its nucleic acid is released inside the cell. ( 1 )
The phage nucleic acid takes over the biosynthetic machinery of the cell to retroflex its won familial stuffs and to synthesise phage specific proteins. When new phage proteins and new phage DNAs have accumulated, they self-assembly into mature phage atoms, the phage specific enzyme ( muramidase ) that dissolves the bacterial cell wall and the phage is released from the bacterium. A individual phage can bring forth 1000000s of offspring at the disbursal of bacteriums cells in civilization. ( 1 ) ( 2 )
4.2 Deadly phage
Bacteriophage are classified as virulent or temperate depending on the nature of their relationship to the host bacteria. Infection of a bacteria by the deadly phage consequences in the lysis of the bacteria and decease releasing freshly replicated phage atoms. One phage can bring forth 100s of offspring within 20 proceedingss under optimum status. The interesting thing about that deadly phage is that the phage that attack one bacterial species do non assail other species. ( 1 ) ( 2 )
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Figure: Scanning Electron Microscopy of T4 cells demoing the Virulent Phages within E coli
Degree centigrades: Documents and SettingsuDesktopactsex.jpgThe bacteriophages shooting their nucleic acids
Degree centigrades: Documents and SettingsuDesktopimages.jpegThe Phage shooting the nucleic acid into the bacteriums
That is a immense advantage to utilize phage as an antibiotic because the phage against E coli bacteriums will merely kill that species and will non impact the others including the normal floral of our human organic structure. So, they will be the most specific antibacterial agent. ( 1 ) ( 2 )
4.3 Temperate Bacteriophage
The temperate phages are different from the virulent one in that they have two possible destinies after infecting the bacteria. Some cause the lysis and decease of the bacteria merely like the virulent phages do but they do hold another alternate result. After come ining the cell, the phage DNA integrates with the chromosome of the host cell. During this province ( prophage ) the cistron look of the phage is continuously by a protein ( represser ) encoded by the phage genome and hence no new phage atoms are produced, the host cell survives and the phage DNA replicate as portion of the host cell. ( 1 ) ( 2 )
4.4 LYSOGENIC BACTERIA
The bacterium which carry the prophage are called lysogenic bacteriums and this phenomenon is called LYSOGENY. The association of the phage and the bacterium is really stable unless the host DNA harm or the exposure to the UV visible radiation occurs. When the DNA harm occurs, the repression of phage cistron is lifted and the lysis occurs and the host cells die. ( 1 ) ( 2 )
4.5 MOLECULAR DETAIL OF LYSOGENY
In Lysogenic mechanism, the cistrons for the lytic procedure will hold to be turned off and this procedure is caused by the phage coded represser cistron. This may be proteins or sometimes anti sense RNA. The represser cistrons can turn off about all the transcriptional induction and so most of the cistron written texts including the indispensable 1s for the lytic mechanism are inhibited. But the represser cistron acts merely onto the few booster cistrons and the cistron for lysis mechanisms in late portion of the procedures are merely indirectly inhibited by the deficiency of early cistron written texts tuned off by the represser proteins or anti sense RNAs.
The early cistron merchandises are needed to trip the subsequent cistron looks and their absence make the whole lysis procedure inhibited. The repressers besides promote their ain written texts to guarantee their maps. The lysogenic province is really stable and merely one in 100000 cell divisions may undergoes lysis by self-generated activation. There are besides many ways to halt the lysogenic province in experimental such as warming in which the represser proteins become denature, handling with the UV UV visible radiation in which the particular system called SOS system of the bacterium is activated. The SOS system is the planetary regulative system which responds to DNA harm. The breakdown merchandises such as oligonucleotides activate the Rec A protein ‘s co-protease activity and this protein in bend inactivate the chief peptidase protein called LexA and so repressers and the lysogenicity province is ended. ( 1 ) ( 2 )
Degree centigrades: Documents and SettingsuDesktop12.jpgFIGURE: THE LIFE CYCLE OF BACTERIOPHAGE SHOWING THE TWO PATHS, LYSIS AND LYSOGENY
There is besides a type of protein called anti-repressor protein and they inhibit the repressers ‘s activities and this anti-repressor synthesis is turned off in the lysogenic cells by the care protein called Mnt protein.
CII cistron: In order to accomplish the stable lysogenic province, both the constitution of the repression of the lytic cistron and the integrating into the chromosome of the cell are needed. CII is a transcriptional activator which is coordinated with the lysogenicity. Within the CII cistron, the cistron called CI cistron which written text needs the CII-dependent booster set up the repression of the lytic cistrons. This booster is activated merely for a short period during lysogenisation and after the repression has been established, the CII cistron is repressed itself.
The CI cistron is so transcribed merely from the autopsy cistron which is the care booster cistron. The another protein called Integrase which is indispensable in integration of the phage nucleic acid with the chromosome is besides transcribed from the CII dependant booster called PI. The pAQ which is besides the CII dependent booster makes an antisense transcript that opposes the Q cistron look. Q cistron ‘s merchandises stimulate the late cistron look and late cistron merchandises which could kill and lyse the cell and they are non made by the CII showing cells and so the CII cistron is so indispensable for the lysogenic province and merely these cells showing CII cistron efficaciously become lysogenic cells. ( 1 ) ( 2 )
LIPPINCOTT ‘s ILLUSTRATED REVIEWS of MICROBIOLOGY 2007 EDITION Bacteriophage in chapter 7 Bacteria familial, cistron transportation, Page 60-61
RICHARD CALENDAR THE OXFORD TEXT BOOK OF “ THE BACTERIOPHAGE ” 2nd EDITION 2006 portion II, Life of Phages, Page 66-104
BACTERIOPHAGE AS AN ANTIBIOTIC
5.1 WHAT PROPERTIES NEEDED AS AN ANTIBIOTIC?
The antibiotic must be effectual in the intervention of infection because of their selective toxicity. That means the drug should kill or consequence against the invading desired being without harming the cells of the host. In most of the instances, this toxicity is merely comparative instead than absolute, necessitating that the concentration of the drug be carefully controlled to assail the micro-organism while still being tolerated by the host. ( 1 )
5.2 WHY PHAGES AS ANTIBIOTIC?
As the PHAGE viruses can infect and kill the bacteriums, they can be used as a drug marks against the Bacteria. Bacteriophage Therapy: Bacteriophage therapy is the usage of lytic phages to kill specific bacteriums as an option to antibiotic. FC: Documents and SettingsuDesktopphages_wikipedia_big.jpgTh
Degree centigrades: Documents and SettingsuDesktopphage_lysis1.jpgBACTERIOPHAGES UDERGOING LYSIS OF THE BACTERIA
The lytic mechanism of the Bacteriophages ensures the effectual antibiotic mechanism of the Phages. The other utile thing is that the specific type of Bacteriophage onslaughts merely the corresponding bacteriums and so the other normal bacteriums will non be affected by the specific Phage Therapy targeted to the aimed bacteriums. So, the Bacteriophages have more specificity than all antibiotics in assailing the bacterium.
5.3 Host Vs Phages
The phages are immunogenic and could originate the immune responses. This consequence may restrict the utilizations of Phages because the Bacteriophages may be destroyed by our immune system even before assailing the coveted mark bacteriums and besides the strong immune responses may trip the allergic reactions and besides the human immune system produces antibodies against the Phages.
Despite these affairs, the good intelligence is that their clinical utilizations reveal merely really few side effects or allergic reaction. The best manner to avoid the sensitisation is to utilize the Phages merely when it is necessary as in the instance of multidrug immune infection and utilizing the Intra Venous IV Administration method.
5.4 BACTERIOPHAGE AGAINST TB
Tuberculosis, one of the oldest diseases known to impact worlds, is caused by bacteriums belonging to the Mycobacterium TB composite. The disease normally affects the lungs, although in up to tierce of instances other variety meats are involved. If decently treated, TB caused by drug-susceptible strains is curable in virtually all instances. If untreated, the disease may be fatal within 5 old ages in more than half of instances. Transmission normally takes topographic point through the airborne spread of droplet karyon green goodss by patients with infective pneumonic TB.
Degree centigrades: Documents and SettingsuDesktopmycobacterium-tuberculosis.jpgThe above diagram is the Mycobacterium Tuberculosis under the negatron microscopy
MULTIDRUG RESISTANT TUBERCULOSIS: This status arises when the Bacteria undergo point mutant in their genome which occurs at low but predictable rates. There are two types of drug resistant
Primary drug resistant: This occurs when the strain infects the 1 who has ne’er been treated before and
Acquired drug resistant: In which, the immune develops during intervention with inappropriate regimen.
Apart from the resistant, some of the patients are non appropriate to give the usual dosage of conventional intervention due to their carbon monoxide bing diseases like nephritic failure, hepatitis or liver failure. ( 1 ) ( 6 )
Mycobacteriophages are the bacteriophages that infect against mycobacterium, the bacteriums doing Tuberculosis and many other diseases like Leprosy. Mycobacteriophages were foremost discovered by the in 1946.They are the dual isolated Deoxyribonucleic acid viruses with non contractile tail belonging to the Siphoviridae household of the Bacteriophage. They besides infect the infective B of the Mycobacterium Tuberculosis complex and now more than 250 mycobacteriophages have been indentified. They are either lytic or temperate. Some mycobacteriophages like DS6A can entirely infect the Mycobacterium TB entirely. Bacteriophages like 13, D 29, TM4, Bxz2 and Chel 2 infect both Mycobacterium TB and other Mycobacterium bacteriums. Their morphological fluctuation is limited but their genomes show excess ordinary familial variableness. The deductions of phages in mycobacterial diseases may be greater than antecedently realized. ( 1 ) ( 2 ) ( 3 ) ( 7 )
Degree centigrades: Documents and SettingsuDesktopmycopha.jpegC: Documents and SettingsuDesktophatfull_jacobs20051202-2.jpg The exposure of Mycobacteriophage under electron microscope
5.6 PHAGE THERAPY
We can utilize lytic phages to kill specifically infective bacteriums as an option to antibiotics intervention particularly for the multidrug immune Tuberculosis. Lack of cognition of bacteriophage biological science and the quality monitoring during the readying of curative stocks had made the therapy hard though the first known curative usage was in 1919.The M. Tuberculosis infections are difficult to handle because the bacteriums are of course immune to many antibiotic. The B may stay in the latent or hibernating province avoiding the action of drugs that require reproduction of the bacteriums. So, the intervention of Tuberculosis requires multiple drugs for extended periods of clip to efficaciously bring around and avoid the drug resistant. The minimum continuance for the intervention is four months with four drugs and so two months with two drugs. The most of import thing is the outgrowth of multi drugs immune strains and that makes the Phage Therapy more interested. ( 1 ) ( 2 ) ( 3 )
Degree centigrades: Documents and SettingsuDesktop467-20090401-TUBERCULOSIS.large.prod_affiliate.91.jpg
Dr Margaret Chan, the director-general of the World Health Organization said, ” The state of affairs is already alarming, and poised to turn much worse really rapidly ” . She and Bill Gates besides stated that they merely had small aid from the modern drugs for the disease that is impacting 9 1000000s people each twelvemonth killing about 2 1000000s of them. The conventional drugs are useless against some strains of TB and they addressed the state of affairs OUT OF CONTROL and A POTENTIALLY EXPLOSIVE ( 5 )
TIM JOHNSON, Mc Clatchy Newspapers
Animal survey: One of the Mycobacteriophage, DS6A, showed decrease in the observed in the lesions in lien, lungs and livers of guinea hogs infected with ( Challenged with ) Mycobacterium TB and the survey showed that the anti bacterial consequence of the phages is at least every bit good as Isoniazid monotherapy. The consequences are assuring in the intervention of TB utilizing phages.
5.7 MYCOBACTERIM INSIDE THE MACROPHAGES: Mycobacteria can shack in the macrophage cells of our immune system. They can even prevail inside the phagolysosome where many bacteriums and pathogens are killed. It was unsure whether the mycobacteriophages can last and retroflex inside the hostile intra-cellular environment with decreased PH. ( 8 ) ( 9 )
5.8 MYCOBACTERIUM SMEGMATIS, THE VEHICLE INTO THE MACROPHAGES: The above job can be solved by utilizing the vector bacteria Mycobacterium smegmatis. The engineering was introduced in 2002. In this engineering, the non-virulent bacteriums Mycobacterium smegmatis act like a bearer into the macrophages.
Degree centigrades: Documents and SettingsuDesktopM-Smeg2.jpgMYCOBACTERIUM SMEGMATIS
Macrophages infected with the Mycobacterium Tuberculosis or Mycobacterium avium were treated with the extra Mycobacterium smegmatis infected with Mycobacteriophage TM4. After they are ingested and destructed by the macrophages, the TM4 phages were released within the macrophages infecting and destructing the infective bacteriums even within the macrophages. The experiments showed the important decrease of both the Mycobacterium TB and Mycobacterium avium. ( 9 )
5.9 D29bacteriophages: This D29 are capable of come ining the Macrophages without the demand of any bearer and they can infect the mycobacteria and kill them efficaciously.
So, the mechanisms of action of mycobacteriophages are wholly different from the conventional drugs and will be so of import in the multi-drug immune instances. The Bacteriophage therapy besides dose non need repetition dosing because the viruses do increase within the mark bacteriums and new virions are released on lysis. The endotoxin may be released into the organic structure after the bacterium has been lysed and this could trip the immune reactions but boulder clay now the clinical usage of bacteriophages revealed merely really few instances of side effects or allergic reaction bespeaking that our human organic structure can truly digest them. The other advantage is that they are cheaper and easier to bring forth than antibiotics. ( 10 )
LIPPINCOTT ‘s ILLUSTRATED REVIEWS of MICROBIOLOGY 2007 EDITION
HORACE T. ADAMS CONTEMPORARY TRENDS IN BACTERIOPHAGE RESEARCH 2009 EDITION
RICHARD CALENDAR THE OXFORD TEXT BOOK OF “ THE BACTERIOPHAGE ” 2nd EDITION 2006
hypertext transfer protocol: //www.microphage.com/technology/phageBiology.cfm
WHO, universe wellness organisation, Global Tuberculosis control Geneva
Harrison text book of Practice of Medicine, USA 6th edition
Fuller, KJ and Hatfull GF 1997 Mycobacteriophage L5 infection of Mycobacterium bovis BCG deductions for phage familial in the slow-growing mycobacterium Mol Microbiaol 26 755-766
Kaufmann SH 2002 Protection against Tuberculosis cytokines T cells and macrophages Ann Rheum Dis 61 Sulll 2 ii54-58
BroxmeyerL Sonsowaka DMiltner 2002 violent death of Mycobacterium by a mycobacteriophage delivered by non virulent mycobacteria, theoretical account for phage therapy of intracellular bacterial pathogen, J infect Dis 1155-1160
Trollip A Albert H and Maskell 2001 Bacteriophage based engineering for the rapid diagnosing and drug susceptibleness testing of TB Am Clin Lab 20: 39-42
Bacteriophage as malignant neoplastic disease hardening agent
6.1 PHAGE DISPLAY is a procedure by which a peptide or a protein is expressed as an exterior merger to a surface protein of a phage atom. The peptide or protein sequence can be deduced from its encoding DNA sequence that resides in the phage atom or in a transductant. Amplification of the Deoxyribonucleic acid of involvement can take topographic point by phage/transductant extension or by polymerase concatenation reaction PCR. By bring forthing big sum of phage atoms, each showing a alone peptide or protein peptide and protein libraries can be obtained. The peptides or proteins interacting with defined molecular marks ( most frequently proteins ) can be isolated from such libraries by enrichments through repeated rhythms of panning. So, the phage show can be regarded as a hunt engine of protein-target interaction. ( 1 )
Bacteriophages are bacterial viruses that have no native affinity to mammalian cells. But we can surprisingly genetically reengineered to expose peptides mergers to surface proteins that can acknowledge and adhere to our mammalian cells. Oligonucleotide sequences encoding for foreign peptides are cloned into phage coat protein cistrons ensuing in combinative libraries of one million millions of different phage ringers exposing encoded peptides on their surfaces. This phage show libraries can be easy screened against assorted biological marks including the integral mammalian cells to give binding molecules with coveted target-specific features. Even the cell-specific peptides indentified through phage show can be used as bringing medieties for building of cistron therapy vectors, liposomes, or targeted drugs to morbid cells in many kinds of upset including the malignant neoplastic disease. ( 2 ) ( 3 )
6.2 PHAGES AGAINST BRAIN TUMORS: Malignant encephalon tumours are really hard to handle because they are heterogeneous, migrate far into next indispensable encephalon normal tissues, are immune to chemotherapy and radiotherapy and besides protected by the blood encephalon barrier. ( 1 ) ( 4 )
Degree centigrades: Documents and SettingsuDesktop617px-Glioblastoma_-_MR_coronal_with_contrast.jpgFig: MRI scan of the encephalon with malignant encephalon tumour
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Figure: The encephalon morbid specimen demoing the encephalon tumour
The phage show might be so much effectual against this fatal status by following agencies.
Bacteriophages can optimise the targeted bringing platforms to malignant encephalon cells utilizing the cell specific peptides.
The emerging of phage investigations for profiling of encephalon tumours in single patients and so doing the personalized intervention based on the profiles of these tumours.
To place the aiming peptides for encephalon malignant neoplastic disease root cells and affinity isolations of the corresponding cell-specific biomarkers for the specific anti-cancer immunotherapy.
To place the peptides that cross the blood encephalon barrier for the effectual drug bringing to the malignant encephalon cells.
The immense advantage of phages over the engineerings aiming the tumours utilizing the natural ligands or antibodies is that phage show can place specific binders to both known and unknown biomarkers every bit good as non immunogenic marks while the other engineerings can merely aim to cognize biomarkers. ( 1 )
6.3 PHAGE-DERIVED DELIVERY SYSTEMS TO BRAIN TUMORS: Using phage show methods, encephalon tumour specific peptides and many researches have been identified the encephalon tumours specific peptides.
Cell line or tumour theoretical account
CMTI ( Cambridge, MA )
U87MG ( human glioma )
Spear et Al
Ph D-12 ( M13, P III ) New England Bio Labs
WAC 2 ( human neuroblastoma, besides binds U373MG human spongioblastoma )
Zhang et Al 2001
Landscape ( f8-1/8 ) Petrenko et al.. 1996
RG2 ( rat Glioma )
Samoylova et Al 2003
Ph D-12 ( M13, P III ) New England Bio Labs
Dgli36, SF767, U87MG, U251MG, U373MG
( human gliomas )
Ho et Al 2004
LL ( and CL6 ( M13, P III ) Mazzucchelli et al 1999 )
Agarwal et Al 2008
EZInet complementary DNA ( human splenocytes phagemid library ) San francisco, CA )
Schluesener and Xianglin 2004
The most of import consequence from these research were that the peptides were glioma specific and could selectively present different ladings including Deoxyribonucleic acid molecules, liposomes, and cytotoxic agents to glioma cells.
The research showed that the bacteriophages themselves can be used to build the cell-specific cistron bringing vectors. The phages transporting eucaryotic look cassettes in their genome have the ability to intercede transgene look in several types of mammalian cells. The other lucky fact is that the phages have been infused in worlds with no evident toxicity or side effects or immune reactions. ( 1 )
6.4 PHAGE PROBES FOR BRAIN TUMOR Profiling: Cell surface molecules that are specific to malignant neoplastic diseases cells are really good marks for anti malignant neoplastic disease interventions. Very selective bringing to these marks will acquire really effectual anti malignant neoplastic disease agents minimising side effects. For this ground, the study of cell-surface markers becomes the cardinal to curative success.
Bacteriophage show can be used for designation of proteins and peptides that recognize specifically and adhere to cell-surface marks. Man-made peptides with cell adhering sequences or whole phage atoms transporting such peptides have the potency for functioning as investigations for profiling cell surfaces of malignant neoplastic disease specimens doing the therapy result more predictable. Unlike antibodies, phage investigations can be besides developed for unknown and non antigenic cell surface markers. Probes which are designed to acknowledge the malignant neoplastic disease specific surface markers can be applied to profile specimens from biopsies or tissues removed at surgery from the patients.
When tumour and normal encephalon subdivisions were probed with the labelled showing glioma-specific peptides, staining was observed on tumour subdivisions but normal encephalon tissue parts showed no staining. This consequence validated the specificity of the phage investigations. ( 1 ) ( 5 )
6.5 PERSONALIZED CANCER TREATMENT WITH PHAGE: The design of the scheme is to fit peptides drugs to the molecular profiles of single tumours. This includes the usage of phage investigations for cell-surface molecular profiling of single tumours followed by cytotoxic interventions formulated on the footing of the tumour profile. Cell-targeting peptides transporting phages are indentified utilizing the surgical specimens and biopsies from multiple malignant neoplastic disease patients and placed into the bank of phage investigations. The bank of ( cytotoxic plus aiming ) peptides is besides created in analogue. Then the molecular profiles of the single tumours are established by utilizing theses phage investigations. The patient particular ( single tumour particular ) combination of the drugs from the bank of peptide drugs is prepared based on the molecular profiles and so the patient is treated with this extremely specific personalized intervention. This method has a great potency for the effectual malignant neoplastic disease killing agents for each person without harming to any normal tissues. ( 1 )
6.6 NEW APPROACH AND TARGETS OF PHAGE DISPLAY AND BRAIN TUMORS
WHY encephalon malignant neoplastic disease recurs? ? Very recent surveies showed that merely a little part of the tumour cells have true tumorigenic and limitless proliferative potency. The encephalon malignant neoplastic disease root cell is the one which shows
Immune to radiation and chemotherapy
Ability to migrate which is the factor straight associated with the encephalon tumour cell invasion and migration into next normal tissues
So, They postulated that even though the chemotherapy can kill the tumour cells, the staying portion of the tumour root cells which is more immune to the therapy is really of import portion of the recurs of the encephalon malignant neoplastic diseases.
The current end is to place the encephalon malignant neoplastic disease cells specific markers and proteins and so selectively aim these root cells without the injury to normal encephalon cells. Phage show can be used as to place aiming molecules and markers that distinguish the encephalon malignant neoplastic disease root cells from their normal parts. ( 1 )
Bacteriophage in DISPLAY, HJORN H LINDQVIST Text book of the bacteriophage, Oxford university 2nd edition 2006 Bacteriophage in Display Page 686
HORACE T. ADAMS CONTEMPORARY TRENDS IN BACTERIOPHAGE RESEARCH 2009 EDITION Phage show in encephalon tumour research pg 237-245
Smith GP Filamentoivus merger phagem fresh look vectors that display cloned antigens on the viron surface Science 1985 228 1315-1317
Harrison text book of Practice of Medicine, USA 6th edition
Souriau C Chiche L Irving R, Hudson, P new adhering specificities derived from Min23 a little cystine-stabalized peptides tissues
WHY HUMAN BECOMES AGED aˆ¦
7.1 Deoxyribonucleic acid telomere and ageing consequence: In each rhythm of reproduction, the telomere ( which is the insistent terminals of the Deoxyribonucleic acid of the chromosome ) becomes shorter and shorter and after a certain period of clip, the reproduction is no longer possible because of the loss of telomere in the chromosome. This contributes to the ageing consequence of the cells. Telomeres are of import in DNA reproductions moving as primers for the lagging strand DNA synthesis.
Degree centigrades: Documents and SettingsuDesktopdnareplication45.jpg Fig: The figure showed the DNA reproduction of dawdling and taking strands. Both taking strands DNA synthesis is non possible because the Deoxyribonucleic acid primers can non synthesise the Deoxyribonucleic acid with the 3 ‘ to 5 ‘ way. But, unluckily, the DNA polymerase can non copy all DNA sequences for the lagging strands because they need infinite to integrate with the female parent strands. So, there will ever be loss of DNA sequences at the very terminal of the chromosome ( DNA ) and the Deoxyribonucleic acid will go shorter with each rhythm of reproduction. But the nature gave us the solution of that job by adding the unneeded sequences ( with no coding part ) at the really ends of the DNAs ( chromosomes ) and we merely loss these sequences called telomere merely without impacting the cryptography parts. But after sufficient repeat of reproductions, these telomeres eventually become loss wholly and impact the Deoxyribonucleic acid and the maps of the cells and the cells becomes loss their regeneration powers and aged and dice.
7.2 Telomerase enzymes are the rearward RNA polymerase enzymes that have the ability to re lengthen the telomere by re-synthesizing the loss DNA ( telomere parts ) with its rearward RNA polymerase activity. With the sufficient activity of the enzymes, the cells will last everlastingly and can reproduce without deceases.
Degree centigrades: Documents and SettingsuDesktopfiles.jpegFig: Showing the telomere and telomerase action
But, the job with the enzyme is that most of the malignant neoplastic disease cells have strong telomerase activity and it may play in carcinoma formation, malignant transmutation ( note that the malignant neoplastic disease cells have the ability to boundlessly retroflex without ageing ) .
My thought and reviews about the enzyme
But I think that we will be able to pull off between these two effectsaˆ¦aˆ¦ over activity of telomerase enzymes in malignant neoplastic disease formation and low activity of the same enzymes in ripening of the cells after a long period taking for the loss of our cell ripening.
I besides think that we can utilize Phage show engineering to selectively heighten the telomerase activities of the selected mark cells. *Example, GI cells or epidermis cells. But, some research stated that the ageing consequence is non chiefly due to the deficiency of the enzyme activity but instead due to the loss of collagen and elastin. Besides some scientists are seeking really hard to cognize the anti-ageing engineering declaring they will detect something astonishing about the engineering to battle the ageing affects of world.