Role of Trace Elements in Duchenne Muscular Dystrophy

Transcription

1 ole of Tace Elements in Duchenne Muscula Dystophy Sanjeev Kuma 1, eena, Mittal 2, Sweety 1 and D. C. Jain 3 1 Depatment of Physics, Cental Medical Physics eseach Laboatoy,D.A.V. (P.G.) College, Muzaffa Naga, U.P. (India). 2 Deptt. of Mathematics, Shi.K.K.JAIN. College, Khatauli, Distt. Muzaffanaga,U.P.India 3 Depatment of Neuology, Safdaganj Hospital, New Delhi, sanjeev1962kuma@yahoo.co.in; ddcjain@gmail.com; eena_math@ediffmail.com ABSTACT Atomic absoption analysis involves measuing the absoption of light by vapoized gound state atoms and elating the absoption to concentation. The incident beam of light is attenuated by atomic vapou absoption accoding to Bee s Law. The estimation of tace elements show a coloful pesentation of diffeent metals. It has been seen and found that the levels of zinc and selenium wee lowe in DMD cases in compaison to healthy contols. The elements such as coppe, calcium, ion, magnesium, potassium, sodium wee found highe than contols. On the basis of statistical analysis we have measued egession and coelation coefficients including multiple coelation coefficients between diffeent tace elements like Na, K, Ca, Mg, Zn, Cu, Fe and Selenium in nomal samples. A tend has been found in coefficient of coelation such as ZnFe > Fe. Se CuZn > NaK > Zn. Se > CuFe with positive coelation and > Cu. Se > CaMg with negative coelation. Coefficient of patial coelation is also calculated and found that a tend has been set up between tace elements and given hee as ZnFe.Cu > ZnCu.Fe > CuFe.Zn > NaK.Ca > CaNa.K > CaK.Na > NaK.Mg with positive coelation and we have found also a negative coelation between some of the elements and a tend is given hee as ZnFe. Se > MgK.Na > MgNa.K Multiple coelation coefficient in nomal healthy peson has been estimated and found that a tend, which is given hee as Zn.FeCu > Fe.CuZn > Cu.FeZn > Zn.FeSe > K.MgNa > Mg.NaK > Na.MgK > K.NaCa > K.NaCa > Ca.KNa. We have also measued coelation coefficients including multiple coelation coefficients between diffeent types of tace elements like Na, K, Ca, Mg, Zn, Cu, Fe and Selenium in DMD samples. A tend has been found in coefficient of coelation such as CaMg > FeZn > NaK > Cu. Se with positive coelation DOI: /jbemi Publication Date: 28 th Octobe 2014 UL:

2 and J O U N A L O F B I O M E D I C A L E N G I N E E I N G A N D M E D I C A L I M A G I N G, V olume 1, Is s ue 5, Oc tobe, 2014 Fe. Se > CuFe > Zn. Se > CuZn with negative coelation. Coefficient of patial coelation is also calculated and found that a tend has been set up between tace elements and given hee as CuFe.Zn > CaNa.K > MgK.Na NaK.Ca > ZnFe. Se with negative coelation and also found a tend of positive coelation as ZnFe.Cu > > NaK.Mg > CaK.Na > MgNa.K. Multiple coelation coefficient in DMD given hee as Mg.NaK. Fe.CuZn > K.NaCa > Zn.FeCu patients has been estimated and found that a tend, which is > Na.KCa > Zn.FeSe > K.MgNa > Cu.FeZn > Na.MgK > Ca.KNa Ou findings suggest that the values of highe levels of tace elements should be adjusted such that these levels must slight below o equal to the nomal limits of the paticula element. The levels of zinc and selenium should be maintained within the nomal limits fo DMD cases in compaison to healthy contols. The elements such as coppe, calcium, ion, magnesium, potassium, sodium may be educed up to the nomal limits in compaison to healthy pesons. Key Wods: Duchenne muscula dystophy, Tace elements, Flame atomic absoption, Multiple and patial coelation coefficiments and egession equations 1 Intoduction Humans ae able to feely move thei bodies about muscles, which is a pecious, complex physiology that should not taken fo ganted. Thee ae so many types of muscles in human body like voluntay and involuntay muscles and smooth and stained muscle fibes that all function in a tight ealm with the nevous system and diffeent chemical eactions. Muscles ae composed of potein in a highly oganized system fom lage goups to small fibes. Muscle units ae sepaated fom othe muscle goups by plasma membanes called sacolemma and the cytoplasm, which is called sacoplasm. Thee ae long potein bundles within the sacoplasm called myofibils. Thee ae so many ATP poducing mitochondia, as well as glycogen and myoglobin. Bundles of paalled myofilaments make up the myofibils whee most of the action takes place. Thee ae contactile poteins in the myofilaments, called myosin, and action. The myosin ae thic filaments and actin thin filaments, when signald, the action and myosin intelock and slide ove each othe to stetch o slide into one anothe fo contaction. They ae signaled fom the nevous system followed by a seies of chemical eactions involving ATP, calcium, sodium and potassium ions. Thee ae many othe poteins involved in the pocess. Abide fom the contactile poteins, thee ae egulatoy poteins called topomyosin and toponin whee act like a switch to detemine when to contact and whee to elax. On the muscle fibe the I band is the space between the myosin (thick) filaments, whee lies only the thin filaments. Thee is a dak disc in the middle of each I band called Z disc. This disc is made of titan. This is connected to sacolemma by the cytoskeleton. As the muscle contacts the I band shinks and the sacomee shotens and as the Z disc s come close togethe pulling on the sacolemma shotening the cell. This is how the muscle contacts? C O P Y I G H T S O C I E T Y F O S C I E N C E A N D E D U C A T I O N U N I T E D K I N G D O M 72 >

3 Sanjeev Kuma, eena, Mittal, Sweety and D. C. Jain; ole of Tace Elements in Duchenne Muscula Dystophy, Jounal of Biomedical Engineeing and Medical Imaging, Volume 1, No 5, Oct (2014), pp One of the most clinically impotant accessoy poteins is dystophin. This is located just unde the sacolemma in the cytoplasm in the aea of the I band. It is poduced by specific genes and links the actin filaments to the potein extacellula matix in the membane known as the dystophin associated potein complex. Elements of the dystophin gene and the potein stuctue have been completely identified. The exact functional ole is still a bit unclea. It is thought that its pimay function is to povide mechanical einfocement to the stuctue of the sacolemma and theeby potecting the membane fom the stess o teaing duing contaction. If dystophin is defective o absent, the membane beaks down, which then substances and molecules like poteins and enzyme leak out the fibe into ciculation. These enzymes and chemicals that leak out ae esponsible fo cetain chemical eactions and necessay to poduce fo muscle contaction. At the same time the extacellula substances leak into the fibe though the boken down membane damaging the fibe and disupting the pocess of muscle contaction and may cause iepaable damage. The absence o abnomality of dystophin esults in a condition known as muscula dystophy (MD). Muscula dystophny is a cippling disease esulting fom mutated genes, which showly wastes away muscle tissue. It has been seen that without dystophin to help to potect the fibe membane keeping it intact, and assisting to ceate enegy, the muscles begin to degeneate and atophy, being eplaced by fat and fibous sca tissue ceating fascia adhesions thoughout the body. It is thought that the majo deteminate of the membane damage would be the level of stess associated with contaction athe than the numbe of muscle actions. Petof, B. et. al. [1] have given thei thoughts to explain why it pimaily affects the peipheal limbs? Muscula dystophies most commonly involve in a genetic mutation in the dystophin genes peventing the poduction of dystophin o limiting the amount in subnomal levels. Geneally in muscle tissue it has a nomal value fo small taes on the sacolemma to occu as the muscle undegoes excessive stain and thee ae small molecules that enhance the natual epai pocess. Howeve in the absence of dystophin, the sacolemma is left unpotected teaing moe fequently and moe easily theefoe muscle degeneation geatly out weights muscle egeneation eventually leading to death and adhesion of the tissue. Emey, A.E. [2] has given his thought egading muscula dystophies, which ae chaacteized by pogessive ievesible degeneation pocess, which esults in weakness and wasting of muscle tissue. The muscula dystophies shae clinical symptoms. These diffe lagely in seveity, age of onset, and distibution of affected muscle tissue. The mechanisms esponsible fo this divegence in pathology ae still not identified in detail. It is useful to undestand the basic anatomy of the neve and muscle disodes. The neve cells have bodies that contain a nucleus. The nucleus contains a genetic mateial, pojection called dendites that eceive infomation fom othe neves, and axons that tansmit infomation to othe neves, ultimately, to muscles. Whethe it be the limb, eyes, bladde, bowel, lungs, o heat, all body movements depend on o egulated by the action of neves on muscle. The bain is made up of billions of neve cells that communicate with each othe by sending infomation acoss inteconnecting axons and dendites, i.e., by synapsing. Some neve cells in the bain have long axons that leave the bain and descend in the spial cod to synapse with the cell bodies of othe neves, the anteio hon cells, in the spinal cod. These cell bodies ae found at each level of the spine. Afte eceiving the infomation fom the bain, U L : 73

4 J O U N A L O F B I O M E D I C A L E N G I N E E I N G A N D M E D I C A L I M A G I N G, V olume 1, Is s ue 5, Oc tobe, 2014 they send these infomations down thei axons (peipheal neves) to communicate synapse with muscles in the limb, head and diaphagm. The axons of limbs peipheal neves synapse with muscle cells acoss meet at a junction called myoneual junction. By definition, neuopathies ae diseases of neves and myopathies ae diseases of muscle. Afte cystic fibosis, myopathies ae the most commonly inheited diseases. Muscula dystophies ae myopathies and tend to pogessive, with ongoing degeneation and egeneation of muscle fibes. Spinal muscula atophy (SMA), amyotophic lateal scleosis (ALS) and polio myelitis ae essentially diseases of the anteio hon cells of the spine. Bach,.J.[3] has given views that people with SMA ae bon with fewe-than nomal anteio hon cells of the spine. Neuomuscula diseases ae diseases of the peipheal neves (neuopathies and anteio hon cell diseases), the myoneual junctions (myasthenia gavis), o the muscles (myopathies) themselves can be undestood with the detailing of these diseases. These conditions cause muscle weakness but do not affect sensoy functions such as the ability to feel objects, see, hea, o smell, no do they affect the autonomic nevous system that contols bladde and bowl function. Evey cell in the human body has foty eight chomosomes. Foty six chomosomes ae autosomes and two ae X o Y sex chomosomes. Foty six autosomes ae twenty thee pais of chomosomes; one in each pai is inheited fom each paent. Ou entie genetic make-up is detemined by the deoxyibonucleic acid (DNA) content of the hundeds of thousands of genes in each of the chomosomes. Defective genes esult in many of the neuomuscula diseases. ecessive diseases equie that the abnomal gene by inheited fom both paents. When an abnomal gene needs to be pesent on the autosome inheited fom one paent only, the disode is called autosomal dominant. Thus, neuomuscula diseases inheited fom defective genes on autosomes ae eithe autosomal ecessive o autosomal dominant disodes. One of about evey adults caies the defective gene fo SMA on one of his o he two numbe 5 chomosomes. Fo a child to inheit SMA, an autosomal ecessive disode, both mothe o fathe must cay the defective gene and tansmit it to the child. Thee is one out of fou chance that a child of paent caies of SMS will inheit the abnomal gene fom both paents and develop SMA. Thee is a fifty pecent chance that the child will be a caie but not develop SMA. Thee will be twenty five pecent chance that the child will neithe be a caie no develop SMA. Childen with SMA must inheit defective genes fom both paents. Fo an autosomal dominant condition like facioscapulohumeal muscula dystophy, on the othe hand, only one paent s affected gene needs to be tansmitted. Each patient must have one affected paent, and thee is a fifty pecent chance of each child inheiting the disease. Neuomuscula disease can also be inheited fom defects on the X (Sex) chomosome. Childen with sex-linked ecessive disodes, such as Duchenne, Becke, and Emey-Deifus muscula dystophies, inheit a single defective gene on the X chomosome fom the mothe and a nomal Y chomosome fom the fathe. Because Y chomosome does not have the genetic mateial to offset the pesence of the defect on the X-chomosome, the child inheits the disease. This is called sex-linked ecessive disode. A female child with a defective X-chomosome fom the mothe and a nomal X chomosome fom the fathe does not develop the disease. She is a disease caie who can tansmit it to he own male childen. When the mothe is a caie, he sons will have a fifty pecent chance of being affected, while C O P Y I G H T S O C I E T Y F O S C I E N C E A N D E D U C A T I O N U N I T E D K I N G D O M 74

5 Sanjeev Kuma, eena, Mittal, Sweety and D. C. Jain; ole of Tace Elements in Duchenne Muscula Dystophy, Jounal of Biomedical Engineeing and Medical Imaging, Volume 1, No 5, Oct (2014), pp he daughtes will have the same chance of being heteozygous caies. Mothe s male elatives may be affected. A suvey has been made and it has been epoted in the liteatue that 70% of the caies of the abnomal gene fo Duchenne muscula dystophy (DMD) actually have some degee of muscle weakness. Thee ae nine kinds of muscula dystophies. These ae classified by the depending on distibution of affected muscle goups, seveity and pognosis, genetic defects and the means of inheitance. Duchenne muscula dystophy is the most common and most sevee. It affects not just all the voluntay muscles but also the heat and espiatoy muscles as well shotening one s life span dastically. This type of dystophy is due to genetic mutations on the twenty thid chomosome, the sex detemining chomosome, being an X linked ecessive o sex-linked ecessive tait. Theefoe the males ae affected fom the gene passed down by thei mothes. Mothe was only a caie. Othe types of dystophies ae those inheited though X-linked ecessive tait ae Becke and Emey- Deifuss muscula dystophy. Becke muscula dystophy is a diffeent mutuated gene. It is located on the same gene locus as Duchenne. This type of dystophy affects the heat tissue in geneal. It is less sevee. It has a longe life expectancy. Autosomal dominate tait may inheit in some diffeent types of dystophies. These ae facioscapulohumeal, distal and occulophayngeal muscula dystophies as well as myotonic dystophy. Facioscapulohumeal muscula dystophy is caused by a missing piece of DNA on chomosome 4. This type of dystophy affects the face, shouldes and uppe ams but late affects cetain muscles of the legs, the abdominal and pelvic gidle leading to exteme lodosis which may equie a wheel chai. This type of dystophy aely affects the heat and espiatoy muscles. It cannot shot the span of life. The most of this type begins at the age of 20 yeas. It speads vey slowly due to apid busts of muscle deteioation. Neuopathies and myopathies such as polymyositis and mysthania gavis ae due to inflammatoy o immunologic disodes that affect muscle. They can also appea in childhood and in many ways esemble inheited neuomuscula diseases. They ae acquied athe than inheited conditions. Thee is no defect in the genes o chomosomes of patient of polymyosis o myasthenia gavis. We ae giving the details of Duchenne muscula dystophy only because the exact theme of the wok is to cay eseach as the appoved and ethically committee cleaed topic. We have chosen this topic to study diffeent aspects of the DMD with spectoscopic tools of Science. Duchenne muscula dystophy is the most common neuomuscula disease.. A case of DMD was fist epoted in 1836 in Italy. It was ecognized and descibed as an inheited disode of boys by the English physician Meyan, E. [4] in The condition was in appopiately named afte Duchenne. Duchenne, G.B.A. [5] was a senio neuologist pacticing in Pais. Duchenne was dispaaged Meyan s wok and fist wote about the condition in Duchenne at fist confused it with ceebal palsy, poliomyelitis, and othe conditions. Although muscle stength may be slightly deceased fom bith. Infants with DMD ae thought to have no physical poblems. It has been epoted in the liteatue that the childen with DMD begin to walk at the age of one and half yeas. Thity pecent childen can walk independently befoe fifteen months of age. Some childen can begin to walk only when they ae thee yeas old. Thee is no coelation found between the late walking and apid pogession o seveity. U L : 75

6 J O U N A L O F B I O M E D I C A L E N G I N E E I N G A N D M E D I C A L I M A G I N G, V olume 1, Is s ue 5, Oc tobe, 2014 Muscle weakness is geneally fist ecognized between ages of thee and five yeas. It pogess symmetically and in a pedictable patten. Childen tend to walk on thei toes. Some childen walk up on thei toes fom outset and ae neve able to walk with thei feet flat. They also tend to waddle and have to climb up thei legs, thighs, and hips when getting up fom the floo o fom the chai. The ability to un, which is geneally achieved by two yeas of age, is not attained. If the childen of DMD ae not popely teated they develop an imbalance in muscle stength at evey joint. Hip flexos emain stonge than hip extensos. Knee flexos emain stonge than knee extensos. The ankle planta flexos emain stonge than ankle dosiflexos. The imbalance in stength at any joint leads to contactue of the joints muscle and soft tissues. The stonge muscles ae shoten. The muscles and thei suounding connective tissues become tight. The joint s ange of motion is deceased. The stonge muscles on one side of the joint stetch the weake muscles on the othe side of the joint. This not only causes contactue of the stonge muscles but also makes the weake muscles get weake even faste, because they ae stetched and no longe at thei ideal length fo contacting. Sevee muscle weakness can pevent ambulation. It has been seen that the childen with DMD and othe neuomuscula diseases lose the ability to walk pematuely because of joint contactues. Mose, H. [6] has studied Duchenne muscula dystophy: Pathogenic aspects and genetic pevention. The autho has put fowad his views befoe the medical science as the patients ae an wheel chai bound at the age of eight to ten yeas. The die befoe the age of twenty yeas geneally. The mutation ate is in the ode of This ate is highe than fo any othe X-linked genetic disease. Thee is no stuctually o functionally abnomal potein known that might epesent the pimay gene poduct. It has no any pathogenic mechanism leading to the obseved biochemical and histochemical alteations been elucidated. The functional defect of the muscula plasma membane is still most attactive. It would be able to explain both the excess of muscula constituents found in seum of patients and caies such as ceating kinase (CK), as well as the excessive calcium uptake by dystophic muscle fibes, which, pio to necosis, could lead to hype contactions uptue of myofilaments in adjacent sacomas and by excessive calcium uptake to mitochondial damage causing cucial enegy loss. Eemey, A. E. [7] and Abood, E. A. et. al. [8] have studied and given thei thought as the incidence of DMD is appoximately 1 in evey The fist symptoms of DMD ae chaacteized by fequent falling, difficulty of getting up fom a standing o lying position and a waddling gait. The calf musculatue is significantly enlaged by hypetophy, fat, infiltation, and accumulation of connective tissue. It has been seen that twenty pecent patients of DMD has a mental impaiment. Affected skeletal musculatue is mainly poximal, and esults in wheel chai dependence duing the ealy teenage. Patients die with cadiae failue as an adolescent. Simands, A. K. et. al. [9] have studied DMD and found that infections, leading to pneumonia as a esult of lack of ventilation ae the cause of death. The cae of espiation may incease the suvivals. Some,H. et. al. [10] have studied DMD and found that a majo chaacteistic of DMD and othe muscula dystophies is displayed of muscle necosis, which esults in high seum levels of the isoenzyme ceatine kinase (CK). The leaks of CK fom affected degeneation myofibes to the blood steam in addition to othe enzymes such as peovative Kinase. C O P Y I G H T S O C I E T Y F O S C I E N C E A N D E D U C A T I O N U N I T E D K I N G D O M 76

7 Sanjeev Kuma, eena, Mittal, Sweety and D. C. Jain; ole of Tace Elements in Duchenne Muscula Dystophy, Jounal of Biomedical Engineeing and Medical Imaging, Volume 1, No 5, Oct (2014), pp Moki, B. et. al. [11] have found that plasma membane defects ae ealy and basic pathologic alteations and ae epesented by lesions of vaious sizes. He also found that thee ae some ultastuctual abnomalities of the membane. Engel, J. et. al. [12] have studied egions nea small lesions contain dilated endocytotic vesicles, wheeas lage lesions habo dilated S Vesicles, iegulaly positioned sacotubula components depending mitochondia, and small clustes of glycogen.gillis, T. N. [13] has studied and found that extacellula calcium can ente the myobibe and cause as imbalance of the calcium homeostasis, which esults in myofibe degeneation. The imbalance of calcium homeostasis might be that degeneation peludes the extacellula calcium influx. Sandi, M.[14], Tidball J. G.et.al.[15] and Matsuda,.et.al[16] have studied the degeneation leading to myofibe necosis, which was initiated via othe mechanisms. The detection of apoptotic nuclei demonstating DNA fagmentation in muscle biopsies fom DMD patients. The absence of the DGC leads to sacolemmal uptue and subsequent leaking of intacellula poteins, which ae unknown to the immune system. Spence, M. J. et. al. [17] have studied DMD and found that the leaking of intacellula poteins attacts and activates cytotoxic lymphocytes and helpe T-cells, which initiate the apoptotic pogam in uptues myofibes. Badley, W. G. et. al. [18] have studied dystophin deficiency and found that this deficiency leads to ievesible myofibe cell death o necosis. The fist necotic myofibe can be demonstated in the neonatal peiod. The myobibe at this stage is single. The necotic myobibes appea in goups of 2-15 myofibes with the incease of age of the patient. Bowman, W. [19] has studied muscula dystophy and found that myofibe necosis is segmental, which means that not the entie myofibe is affected by necosis. The basal lomina suounding the necotic myofibe is not affected. It emains empty shell. The affected myofibes contain membane attack complexes, which ae the esult of complement activation. The insetion of this in the sacolemma ceates holes and causes cell-lysis. The inteests of bevity and accuacy ae both popely seved by a simple statement that we do not know the etiology of DMD. Thee has been so much discussion in the last few yeas challenging the taditional concept of muscula dystophy as a pimay illness of the muscle. Muphy, D. L. et. al. [20] have given thei views egading the seach fo abnomalities in catecholamine metabolism and it was acceleated by the findings that thee was a educed initial ate of accumulation of seotonin in platelets fom the DMD patients. Paulson, O. F. et. al. [21] have studied blood flow in muscle of DMD patients and could not find any abnomality. Jeusalem, F. et. al. [22] have studied electon micoscopic and mophometic studies of the capillaies in muscle fom patients with the disease have been unevealing. Mc Comas, A. J. et. al. [23] have suggested that the muscle fibes in Duchenne muscula dystophy wee not being lost in a andom fashion but disappeaing in goups associated with the loss of moto units. They also poposed that DMD might have its oigin in the malign influence of a sick moto neuon upon the muscle fibes. Panayiotopoulos, C. P. et. al. [24] and Ballantyne, J. P. et. al. [25] have studied DMD and could not find any abnomality in the muscle fibes. The disease may be associated with an abnomal neual influence. U L : 77

8 J O U N A L O F B I O M E D I C A L E N G I N E E I N G A N D M E D I C A L I M A G I N G, V olume 1, Is s ue 5, Oc tobe, 2014 Matheson, D. W. et. al. [26] have studied the changes in the behaviou of diffeent cell membanes and citically descibed the DMD and concluded that an abnomal suface defomation of eythocytes poduced by washing the ed blood cells of DMD patients in saline. Miale, T. D. et. al. [27] and Mille, S. E. et. al. [28] have studied DMD patients and could not attain the esults of Matheson, D. W. et. al. [26] and poposed that it may be a non-specific outcome of Matheson, D. W. et. al. [26] wok. The behaviou of some of the enzymes, which ae associated with sacolemmal o eythocyte membanes ae anomalous. Bown, H. D. et. al. [29] have studied the activity of Na, K, ATPase of eythocyte membanes in DMD found that this is stimulated by ouabain, athe than being inhibited as in the nomal situation. Pete, J. B. et. al. [30] have shown that a facto pesent in the seum of DMD patients may contibute to the alteation given by Bown, H. D. et. al. [29]. oses, A. D. et. al. [31] have studied endogenoces phospnoylation of a goup of membane poteins. This potein activity is called as potein Kinase activity and it is found abnomal high in eythocyte membanes. Mawatai, S. et. al. [32] have studied adenyl cyclase which is an enzyme and it is found to be abnomal. It is associated with sacolemmal membanes. The enzyme is stimulated by epinephine and sodium fluoide but in DMD patients this stimulation is less than nomal.most of the effots in caie detection have been diected to the detection of subclinical abnomalities. Muscle biopsy can be done. Peace, G. W. et. al. [33] and oy, S. et. al. [34] have studied muscle biopsies in DMD patients and one may see foci of necotic fibes, moth eaten and wholed changes in intemyfibilla netwok patten. The intenal nuclei ae in abnomal numbe. These changes wee also found in nomal healthy pesons. These patients ae not caies and the gasto enemius muscle is biopsied. Thei changes ae found in patients who ae at the isk. This is the stong pesumptive evidence of the caie state. Ionasescu, V. et. al. [35] have found an abnomality in ibosomal potein synthesis of DMD patients and found that isolated ibosomes wee synthesized and ae in inceased amounts of collagen. Ionasescu, V. et. al. [36] have also found simila situations in the caies of DMD patients. ibosomes wee found to synthesize abnomally highe amounts of noncollagenous potein as well as collagen. oses, A. D. et. al. [37] have studied and concluded that the inceased ability of the eythocyte membane to phosphoylate potein is found in caies of the illness. The mean values fo goups of definite, pobable and possible caies wee found to be simila and diffeent fom the mean value fo matched healthy contols. The mutation ate must be vey low. If thee wee non-caie mothes in the possible and pobable goups, thei nomal values of potein Kinase should have educed the mean value of these two goups compaed to the definite caies. Hobbins, J. C. et. al. [38] have studied an inteesting concept of the detection of DMD patients. They have pefomed amneucontesis at the fifteen weeks of pegnancy and a detemination of the sex of the fetus. If the fetus is female, the child will not suffe the clinical symptoms of DMD. If the fetus of the examination uteus is male, then mothe may elect to have the pegnancy teminated. It is the question of emotional aspects of the mothe and can be left to decide what should be done? Analysis of amniotic C O P Y I G H T S O C I E T Y F O S C I E N C E A N D E D U C A T I O N U N I T E D K I N G D O M 78

9 Sanjeev Kuma, eena, Mittal, Sweety and D. C. Jain; ole of Tace Elements in Duchenne Muscula Dystophy, Jounal of Biomedical Engineeing and Medical Imaging, Volume 1, No 5, Oct (2014), pp fluid CPK is not a useful test to detect whethe a male fetus has the illness o not. It has become feasible to obtain fetal blood at the eighteen weeks duing fetuscopy penatal diagnosis of the illness. Bodensteine, J. B. et. al. [39] have studied calcium in muscle of DMD patients and found that thee is a tendency of calcium level to be on the highe side. Betoini, T. E. et. al. [40] have studied calcium and magnesium content in muscle of DMD patients and found that the levels of calcium wee highe and levels of magnesium wee lowe. Some of the eseaches have applied Feeze factue technique (FFT) to study the muscle membane and splits the suface membane, peeling the oute suface away fom the inne suface along a line, which passes though the inteio of the membane. Both the faces can be looked at unde the electon micoscope and these faces ae found to be coveed with paticles of vaious types, some of which ae aanged with a ectangula distibution. These ae called squae o othogonal aays. Schotland, D. L. et. al. [41and 42] applied FFT to the DMD patients and found that thee is a significant diminution in the numbe of othogonal aays. Peluchetti, D. et. al. [43] have studied this technique and suggested that thee may be a decease in the intemembanous paticles on both sufaces. The cell suface membane is not homogenous. Thee ae so many poteins and othe compounds embedded in cell membane, which seving diffeent aspects of membane activity. The glycopoteins and glycolipids of cell membanes pefom impotant functions including a ole in the ecepto sites. Thee ae some pobes to identify these constituents by vitue of the selective binding of the pobe to a specific compound. One such pobe is Concanavalin A. This pobe is a membe of goup of plant poteins called lections which have a specific affinity fo suga esidues. Concanavalin A binds specifically with mannose, glucose, and fuctose esidues. Heimann-Patteson, T. D. et. al. [44] have studied the patten of Concanavalin A binding in DMD patients and found that it was patchy and iegula. It was egula and have a good patten in nomal muscle. They have also found an abnomality in the cell membane. Booke, M. H. [45] has witten in his book on muscula dystophy as if DMD is a pimay abnomality in the muscle, then we would suppose to the change might be epoduced by cultuing muscles cells fom patients with the illness. Ionasescu, V.et.al.[46] and Thompson, E. Jet.al.[47] have studied DMD patients and found some abnomalities in the gowth patten which ae able to change in the isoenzyme patten of CK in the myotubes. It has also been noticed that this gowth patten is suspiciously nomal in the cultued muscle. FFt does not eveal any abnomality in the inte membanous paticles. ottman, S. M. et. al. [48] and Mawati, S. et. al. [49] have found that the cultued cells seem to behave nomally electo-physiologically. Cei, C.A. et. al. [50] have found that thee is abnomality in adenyl cyelase. It was not the same as that found in the dystophic muscle itself. It is believed that the possibility of hypothetical membane abnomality might affect othe tissues then muscle alone has been aised. DMD is a genetic disease and diffeent cell suface membanes shae many simila popeties. owland, L. P. [51] has studied the elatively in exhaustible supply of membane in the ed blood cell and this has led to a legion of studies. Falk,. S.et.al.[52], Fische, S. et.al.[53], oses, A. D et.al.[54], Maby, M. E. et.al.[55]and oses, A. D. et.al[56] have studied the ed cell membanes have been both inculpated and exculpated of all of the following such as changes in size, shape, defomability and othe physical popeties, abnomalities in U L : 79

10 J O U N A L O F B I O M E D I C A L E N G I N E E I N G A N D M E D I C A L I M A G I N G, V olume 1, Is s ue 5, Oc tobe, 2014 diffeent ATPases, changes in Calcium tanspot, abnomalities of electolyte content and pemeability, abnomal phospnoylation of potein, abnomalities of adenyl cyelase. Athu, H. et. al. [57] have found that thee is an abnomality in low density lipopoteins in the plasma in DMD childen leading to a suggestion that thee tanspot capacity was impaied. Fiboblasts cultued fom the skin of patients with the illness have also been havested in an attempt to detect abnomalities. A deceased adhesiveness between fiboblasts has been descibed using a technique in which the cells ae encouaged to collide with each othe and the stickiness is measued in such collisions. The adhesiveness has been found to be educed in the fiboblasts fom Duchenne patients. Thee is an ovelap between the nomal healthy contols and the DMD patients. Cultued skin fiboblasts also contain CK, which might make this a useful tool fo studying the enzyme in disease states. Mineals ae impotant to ou health. These ae inoganic, chemical is elements not attached with cabon atom. Mineals and tace mineals can be diffeentiated easily. These tace mineals ae called tace elements. If the cellula body equies less than 100 mg of eleven mineals is labeled a tace element and if accepts moe than this level is labeled a mineal. Tace elements ae equied in quantities of few milligams o micogams pe day. A study of elationships of mineals with human health is vey necessay and impotant. A balance of the level fo mineals in evey ogan, tissue and cell of the human body may lead to a good health. Mineals compise only a faction of total body weight. These ae cucial fo many functions of the human body. These include tanspoting oxygen, nomalizing the cental nevous system (CNS) and simulating gowth, maintenance and epai of tissues and bones [58]. omeyn, M.[59] has given a statement that the egulation of tace elements balance in the body is essential to suvival. Evey cell is a living oganism and we must ty to maintain its intenal envionment. The mobility of tace elements acoss cell membanes, between the exta-cellula and inta-cellula fluid foms the basis fo the body s most pimay functions. The electical activity may be initiated; heat beats, neve cells signal, muscles may espond, blood vessels may tighten o elax. Wate balance should be maintained. Most of the tace elements found in the tissues and body fluids ae also pesent in the blood. Some of the metallic elements have a specific metabolic function and the pesence of these is not meely the esult of contamination fom ou envionment. The tace elements, which ae biologically active poduce thei effects though enzyme systems. Metalloenzymes and metal potein complexes ae two systems. The fome has a fixed amount of specific metallic ion pe molecule of potein addition of an agent, which binds the metal inactivates the enzymes and the late is a lage goup of poteins loosely bound to metal. The metals may substitute fo each othe with fluctuating degees of affinity and can be taken out fom the body by the pocess of dialysis. Some of these complexes may have enzymatic activity only. ests of these ae concened with tanspotation pocess only. Heing,W.B.et.al. [60] have shown that chelation affects the activity of enzymes. Khushid, S.J.et.al.[61] and Suhaila,. et.al.[62] have shown that the essential metals at tace levels play a ole in the human body and can cause some diseases when pesent beyond the limit of nomal concentation. C O P Y I G H T S O C I E T Y F O S C I E N C E A N D E D U C A T I O N U N I T E D K I N G D O M 80

11 Sanjeev Kuma, eena, Mittal, Sweety and D. C. Jain; ole of Tace Elements in Duchenne Muscula Dystophy, Jounal of Biomedical Engineeing and Medical Imaging, Volume 1, No 5, Oct (2014), pp It has been seen that the tace elements occu in the tissue of animals, plants and mico-oganisms in vey low concentations. The concentation of these elements change highly among many elements. Thei concentations ae also diffeent among vaious living oganisms and thei pats. These diffeences ae seen in nutitionally essential elements and also in those elements, which ae associated with no vital function. The tace elements have divegent oles to play depending upon thei chemical fom o combination and thei position in the body fluids and tissues. The functional foms of the tace elements and thei chaacteistic concentations ought to be maintained with naow limits if the functional and stuctual integity of the tissues is to be safeguad and the gowth, health and fetility of the individual have to be unaffected. Continued ingestion of diets that ae deficient, imbalanced, o excessively high in a paticula tace element invaiably induces changes in the functioning foms activities, o amount of these elements in the tissue o fluids, so that they fall below, o ise above the nomal and pemissible limits o anges. Biochemical distubances in these situations develop along with physiological functions, which ae influenced and stuctual disodes may aise due to the change with elements. Kuma, S.[63] has epoted in his Ph. D thesis that the degee and duation of the dietay deficiency o toxicity, age, sex and species ae impotant factos associated with the tace elements. It has been seen that food is the basic need of life of a living oganism. Eveybody eats food whethe it is vegetaian o non-vegetaian. Scientists wee vey cuious about the food they consumed. Food passes in the body and affects the body. Food is the substance taken into the body that will help to meet the body s need fo enegy. It helps in maintaining good health, health gowth, gowth and epoduction. Poteins ae impotant and vital to any living ceatue. These ae the impotant constituents of cells and tissues of the human body. Poteins fom the impotant component of muscles and othe tissues and vital body fluids like blood, ceebospinal fluid (CSF) and synovial fluid in human joints. The poteins in the fom of enzymes and homones ae concened with a wide ange of vital metabolic mechanisms in the body. They supply the body building mateial and make good the loss which occus due to wea and tea. Poteins as antibodies may help the body to defend against infections. Poteins ae one of the most impotant nutients equied by the body has to be supplied though the diet we consume. The dietay poteins ae boken down into amino acids and absobed and ae used by the body fo vaious functions like tissues building, eplacement of poteins depleted, etc. Amino acids ae used fo poviding enegy to the body. Fat is an impotant component of ou diet. It seves a numbe of functions in the body. Fat is a concentated souce of enegy and it supplies, pe unit weight moe than twice of enegy funished by eithe poteins o cabohydates. Fat also impats palatability to a diet and etads stomach emptying time. Fat in the diet can be of two categoies, the visible and invisible. The visible fats ae those deived fom animal fats like butte, ghee which ae solid fats and those deived fom vegetable fats like goundnut, mustad, coconut, sunflowe oil, till, which ae liquid fats. Hydogenated vegetable oil known as Vanaspati is a solid fat. These fats ae tiglyceide of fatty acids, both satuated and unsatuated. Animal fats like ghee and butte contain Vitamin A and D. These vitamins ae not pesent in vegetable oils. U L : 81

12 J O U N A L O F B I O M E D I C A L E N G I N E E I N G A N D M E D I C A L I M A G I N G, V olume 1, Is s ue 5, Oc tobe, 2014 Some amount of fat is pesent in othe foods like ceeals, pulses, oil seeds, milk, egg, meat, etc. This type of fat contibutes to the total fat and essential fatty acids content of diet depending upon the foodstuff pesent in the diet. Nuts, oilseeds, soyabean, avocado, pea and animal foods have a highe amount of invisible fat. Mineals and tace elements ae ciculating in the human body. Some of these elements fom pat of body stuctual component and some othes act as catalytic agents in diffeent body eactions. Bones and skeleton ae made up of calcium, magnesium and phosphous and ion, which is a component of blood. Mineals like zinc, molybdenum, coppe, manganese and magnesium ae eithe a stuctual pat o activate a lage numbe of enzyme systems. Iodine is a pat of homone, thyoxin. Sodium, potassium ae impotant elements pesent in fluids within the outside the cells and along with ions like chloide, bicabonate and cabonate keep wate and acid base balance. Evey individual absobs mineals in a slightly diffeent way, called biochemical individuality. Those nutients that have not been tansfeed though the intestinal mucosal cell to ente the ciculation have no meaning egading the study of the elements. The vaiety of nutients fom oganism s envionment that have been made available by absoption must be tanspoted though the ciculatoy system to the aqueous micoenvionment of the cells. They may seve the pupose of paticipation in diffeent metabolic pocesses in the cells on which the life of the total oganism depends. The absoption of mineals is dependent on many diffeent factos elated to the mechanism of human body. May,.L.A. et.al [64] have epoted that the addition of vitamins and mineals to food can be effective public health intevention to coect inadequate intake of nutients in both the geneal society. 1.1 Diffeent types of Tace Elements ciculating in Human system: Thee ae ninety two elements found in natue. We have some additional twenty two elements also. It has been epoted in the liteatue that thee ae hundeds of isotopes of the elements exist. We may say the any one of which may play an undiscoveed ole in human health. Alaxande,G.S. [65] epoted that though geophysical foces, mixing of the eath s cust with wate may povide vitually evey mineal fo ou body, which equies to maintain health. The wate is compatable with moe substances that any known solvent. It is an ideal medium fo tanspoting nutients in the cells fo the chemical eactions of cellula metabolism to take place. Human body contains lot of element out of 25 ae divided into mainly thee categoies such as majo, mino components and tace elements. Pike,.L.et.al.[66] have given a statement that the majo components, which make up 96% of human body ae composed of oxygen, nitogen, cabon and hydogen. Majo elements found in the biological molecules such as poteins, nucleic acids, fats and cabohydates and ae capable to make-up the body. Hydogen ions ae vey impotant fo ou cells to make ATP o enegy. Wate is also a medium fo all the chemical eactions taken place in the human body. Wate is a mixtue of oxygen and hydogen atoms. Mino components make up 4% of ou body. They may include calcium, phosphous, potassium, sulfu, sodium, chloine and magnesium. Some of these ae necessay fo chemical pocesses to un coectly. Ou muscles need calcium in ode fo them to twitch popely. Enough calcium is equied to ou body. C O P Y I G H T S O C I E T Y F O S C I E N C E A N D E D U C A T I O N U N I T E D K I N G D O M 82

13 Sanjeev Kuma, eena, Mittal, Sweety and D. C. Jain; ole of Tace Elements in Duchenne Muscula Dystophy, Jounal of Biomedical Engineeing and Medical Imaging, Volume 1, No 5, Oct (2014), pp Tace elements make up 0.1% of ou body. They include coppe, zinc, selenium, molybdenum, fluoine, iodine, manganese, cobalt, ion, lithium, stontium, aluminum, silicon, lead, vanadium, asenic and bomine. These elements ae essential fo ou body to function popely. Zumkley, H. [67] stated that clinical expeimental and epidemiological studies indicate that a lage numbe of tace elements may be involved in the etiology of diffeent human body disodes. Both inceased levels of any tace element o evese of this can influence the development of disease. 2 Tace Elemental Effects on Human Health A vey high concentation of the tace elements may pove toxic. Depletion in the concentation of essential tace elements may lead to ceate so many diffeent metabolic instabilities due to dysfunction of enzyme. Diffeent types of metabolic changes in human body ae accompanied by changes made in the concentation of one o moe tace elements in some body fluids, such as blood seum o plasma. The tace elemental analysis fom human hai, nails, and body fluids such as plasma, seum and ceebospinal fluid has been stated in the liteatue as the best pedicato. 2.1 Tace elemental ole in human Body: We ae giving the desciption of some of the impotant tace elements hee as Coppe: The pesence of coppe in animals and plants is vey necessay. Coppe is most abundant tace element in human beings. This element is caied mostly in the bloodsteam on a plasma potein. We may call it is ceuloplasmin. This element is fistly absobed in the gut and tanspoted to the live bound to albumin. It is found in a vaiety of enzymes, including the coppe cente of cytochome C oxidase. Human body needs coppe fo nomal gowth and health. It is said that coppe is equied to help body use ion. This element is vey impotant fo neve function, bone gowth and to help body use suga and it is a component of a cofacto fo diffeent enzymes, appoximately. Fifty enzymes ae equied coppe to function popely. Davis, I.J.J. [68] has given a statement that a young peson equies one hunded to one hunded fifty milligam of coppe daily. Coppe in highe concentation is found in live, hai, muscle and lung Zinc: Zinc Zn) is a necessay tace metal of suitable impotance in the dietay management of many human body complex disodes. A young peson has 3 gm of zinc. This tace element is found in all ou cells. This element is essential fo all foms of life. Many poteins contain stuctues called zinc finges may help to egulate genes. The distibution of zinc in human tissues is well documented in the liteatue. Live, kidney, bone, etina, postate and muscle appea to be vey ich in zinc. Buns,.. et.al.[69] have studied and found that most of the ed cells zinc is tightly bound to enzyme cabonic anhydase. It is exchangeable with plasma zinc. Whole of plasma zinc is found with poteins. Sixty pecent zinc is bound to albumin and thity pecent has binding with alpha 2 macoglobulin and the emainde with low molecula weight poteins including amino acids. Pasad, A. S. et. al. [70] have shown that zinc is bound to amino acids and seum poteins. Histidine, glutamine, theonine, cystine and lysine showed the most makable effects. The amino-acid-bound U L : 83

14 J O U N A L O F B I O M E D I C A L E N G I N E E I N G A N D M E D I C A L I M A G I N G, V olume 1, Is s ue 5, Oc tobe, 2014 faction of zinc may have an impotant ole in biological tanspot of zinc. The stable zinc content is highe in albumin faction. Simila concentations of zinc wee found in the α, β and γ-globulins. These ae faction of immunoglobulin G. Kelin, D. et. al. [71] have shown the fist demonstation of a specific biological function citically dependent on the pesence of zinc. They have also shown that cabonic anhydase contains zinc and it is essential to the mechanism of action. iodan, J. F. et. al. [72] have epoted in the liteatue that thee ae moe than seventy zinc metalloenzymes. Zinc metalloenzymes exhibit divesity both of catalytic function and of the ole played by the metal atom. It is evident that zinc in small quantities is pesent in vaious cell membanes. Most of the membanebound zinc is linked to a distinct macomolecule constituent lipopotein faction. Chavapil, M.et.al.[73] have shown that the effect of zinc and othe metals on aggegation of platelets and elease of H 3 seotonin activated eithe by collagen o epinephine. Pasad, A. S. et. al. [74] have shown that a syndome of ion deficiency anaemia, hepatosplenomegaly and dwafism has been found to be associated with malfunctioning of zinc metabolism. Walavens, P.A. et.al.[75] have studied and found that the othopedic and CNS have elatively slow zinc uptake. This element emains tightly attached fo a lengthy span of time. The deficiency of zinc leads to gowth etadation o failue, lesions of the skin and its appendages, and impaied epoductive development and function. Psychological distubances ae epoted in individuals suffeing fom acodematitis enteopathic. Henkin,.I. et.al.[76] have studied zinc induced deficiencies in human beings and these ae associated with neuological symptoms such as depession, poo concentation, nevousness and moodiness. Desti, I.E. [77] has studied the hippocampus, which is ich in zinc. Zinc deficiency is associated with leaning and memoy defects simila to behaviou syndomes esulting fom destuction of the hippocampus. Acute oal zinc toxicity has been epoted to poduce dowsiness and somnolence Ion: Faibanks, V. E. F. et. al. [78] have epoted that the theapeutic indications fo ion duing the oman ea included alopecia, acne, dematitis, wounds, hemohoids, gout, pulmonay diseases, diahoea, vomiting, weakness, edema, feve and cystitis. McCance, et. al. [79] have epoted that once the ion was absobed by the human body, its excetion was vey minimal and was not contolled eithe by gastointestinal tact o by the kidneys. The plasma ion was in equilibium with tissue ion. The level of plasma ion influenced the ate of ion absoption. The intestine egulates the ate of ion absoption. Ganick, S. [80 adopted and extended the hypothesis of feitin as the mucosal egulato of ion absoption. This hypothesis was used and named as mucosal block theoy. Mooe, C. V. et. al. [81 have shown that the plasma ion as the majo fom of tanspot ion. Flavopotein enzymes of ion wee discoveed by Mahle, H.. et. al. [82] and ichet, D. A. et. al. [83]. Fishe, H.et.al. [84] have synthesized fou feopotopophyin (heme) moieties. Ingam, D. J. E. et. al. [85] have extablished a thee-dimensional pictue of molecule with its fou attached hemes. They have studied the natue of the bond between ion and globin. Now ion is C O P Y I G H T S O C I E T Y F O S C I E N C E A N D E D U C A T I O N U N I T E D K I N G D O M 84

15 Sanjeev Kuma, eena, Mittal, Sweety and D. C. Jain; ole of Tace Elements in Duchenne Muscula Dystophy, Jounal of Biomedical Engineeing and Medical Imaging, Volume 1, No 5, Oct (2014), pp stabilized in the feous state completely, which allows it to be evesibly bonded to oxygen. Due to this mechanism, hemoglobin woks as an oxygen caie. The ion in human seum was studied by Holmbeg, G. C. et. al. [88] Schade, et. al. [87] have shown that the tace element ion may be bounded completely to tansfein. Lipschitz, D.A. et.al.[88] and Simes, M.A.et.al.[89] have established a coelation between seum feitin, concentation and stoage ion Magnesium: Aikawa, J. K. [90 and 91] showed in the studies elated to magnesium and found that magnesium is associated with so many diffeent biological pocesses. Magnesium is pesent in a small concentation in all cells and it is necessay fo cellula metabolism. This tace element is also pesent in bone along with calcium. It shaes so many popeties of calcium so fa as absoption and metabolism and tissue distibution ae concened. Widdowson, E. M. et. al. [92] have studied the magnesium content in human body and found a ange of magnesium, which stated fom 22.7 to 35.0 meq/kg weight of tissues. Magnesium eaches us in many foms. These ae magnesium amino acid chelate, magnesium cabonate, magnesium oxide, magnesium oxide dolomite and magnesium sulfate. Seventy pecent of human body magnesium esides in bones and teeth.this element is essential to the functioning of human body because it tansmits neve impulses, causes contaction of muscles and integal to healthy development of bones and teeth. Watelow, J.C.[93], Clasen,H.G.[94] and AL-Ghamadi,S.M. et.al.[95] have shown that the human body contains about 760mg of magnesium at the time of bith. It goes upto 5 gm at the age of 4-5months and it eaches upto 25 g when a body attains an age of adult. About 30 to 40% magnesium is found in muscles and soft tissues. One pecent is found in extacellula fluid, and the emainde is in skeleton. If we ae going to conside the paticula impotance with espect to the pathological effects of magnesium depletion. We have to take into account the ole of this element in egulating potassium fluxes and its involvement in the metabolism of calcium. Weste,P.[96] has shown that about 1% of magnesium is found in the extacellula fluid, inside cells, magnesium may be bound to phospholipids. It is vey impotant to note that inceasing dietay potein intake may incease magnesium equiements because high potein intake may decline magnesium etention.it is advisable that to completely undestand magnesium function, it is vey impotant to exploe magnesium s elationship with calcium and potassium. Magnesium and potassium also have a close elationship. Magnesium is equied fo the function of sodium and potassium pump. If a magnesium deficiency occus, then pumping of sodium out of the cell and pumping potassium into the cell may be impaied. Kasne,B.[97],Fuukawa,Y.et.al.[98],Stak,G. et.al.[99] and Haigney,M.C.P. et.al.[100] have studied and shown an effect of the level of magnesium egading heat attack. Alcoholism, enal disease, diabetes mellitus may all cause a deficiency of magnesium. Nausea, vomiting, anoexia, muscle weakness, muscle weakness, muscle spasms and temos ae the few symptoms and signs elated to deficiency of magnesium. Magnesium has an effect on CNS. It is used fo a bette sleep. This element may be used to calm iitated and ove excited neves. This is useful with epileptic attacks, convulsions in pegnant women U L : 85