Abstract:
A specimen of an amniotic fluid is obtained and analyzed by GC/MS in order to generate a comprehensive metabolic profile. The profile is analyzed by comparing the levels of metabolites with normal levels of those compounds. Specific treatment is then prescribed for the metabolite levels that differ from the norm. These metabolites that are present in different levels than a normal specimen may be indicative of chromosomal abnormalities such as Down Syndrome. The method of the present invention is used to model the complex problem of a chromosomal abnormality as the sum of several simpler problems that may be treatable. The comprehensive metabolic profile is used to detect chromosomal abnormalities, to suggest treatments for fetal chromosomal abnormalities, and to monitor their effectiveness.

Description:
RELATED INFORMATION 
       [0001]    This application is a continuation of co-pending application Ser. No. 09/499,006 filed on Feb. 4, 2000 and incorporated herein by reference. 
     
    
     FIELD OF THE INVENTION 
       [0002]    The present invention relates to the diagnosis of biochemical abnormalities found in a fetus. Specifically, the present invention is a comprehensive metabolic profile of an amniotic fluid specimen to diagnose and prescribe treatment for chromosomal disorders, preferably Down&#39;s Syndrome, that may be present in a fetus. 
       BACKGROUND 
       [0003]    Chromosomal abnormalities are forms of birth defects that occur in 2 to 3 out of every 1000 births. Chromosome abnormalities may involve duplications or defects in a whole chromosome or a portion of one or more chromosomes. The most common chromosome abnormality is Down Syndrome. In Down Syndrome, each cell contains three copies of chromosome number 21, a condition referred to as trisomy 21. A result of trisomy 21 is a 50% increase in gene dosage for each gene on chromosome 21. At least some of the abnormalities seen in Down Syndrome can be attributed to excessive gene dosage. Down Syndrome is a complicated disorder, affecting many aspects of physiology. Since affected individuals all have mental retardation, it is a significant disability. 
         [0004]    Less common chromosomal disorders involve other chromosomes. Some chromosomal disorders result from a deletion or a duplication of a portion of a chromosome. Other chromosomal disorders result from an absence of an entire chromosome. For example, an absence of the X chromosome results in Turner Syndrome, referred to as monosomy X. 
         [0005]    The relationship between chromosome abnormalities and physical or metabolic disorders in children is not always clear. Down Syndrome is the most common and most thoroughly studied chromosome abnormality. This has spurred some authors to look for treatable or correctable abnormalities in children with Down Syndrome. Le Jeune suggested that abnormalities of purine synthesis, thyroid metabolism, and perturbations of enzymes, specifically copper/zinc superoxide dismutase, phosphofructokinase, and cystathione beta synthase, were related to Down Syndrome. Le Jeune, J.,  Pathogenesis of Mental Deficiency in Trisomy  21, Am. J. Med. Genetics Suppl. 7: 20-30 (1990). Additionally, Patterson and Ekvall suggested that a number of vitamins, minerals, and other metabolic abnormalities contributed to Down Syndrome. Patterson, B. and Ekvall, S. W.,  Down Syndrome in Pediatric Nutrition in Chronic Diseases and Developmental Disorders: Prevention Assessment and Treatment , Oxford Univ. Press, New York, 149-156 (1993). These theories have not been confirmed by other authors. 
         [0006]    Several treatments have been proposed to address Down Syndrome. Turkel advocated the use of “orthomolecular” therapy. Turkel, H. B.,  Medical Amelioration of Down Syndrome Incorporating the Orthomolecular Approach , Psychiatry 4(2): 102-115 (1975). Harrell, et al. suggested supplementation with megadose vitamins. Harrell, et al.,  Can Nutritional Supplements Help Mentally Retarded Children? , PNAS USA 78(1): 574-578 (1981). As reported by Patterson and Ekvall, attempts to verify the effectiveness of the aforementioned treatments have proven unsuccessful. The difficulty of verifying the effectiveness of proposed treatments is compounded since researchers have not confirmed many of the proposed abnormalities. Also, some abnormalities could interact with other abnormalities. Folate metabolism, for example, is also involved in purine synthesis. Consequently, there is a need for a method to comprehensively analyze a wide panel of metabolites in order to create a complete biochemical picture of a given chromosomal abnormality. Using this method, corresponding treatment may then be identified and prescribed as a result of the complete biochemical picture. 
         [0007]    The treatments of Turkel and Harrell et al. might be the right treatment given at the wrong time. Treatment, to be effective, oftentimes must be administered at the proper time. For example, supplementation of folate before and during neural tube development often prevents spina bifida. However, folate supplementation given after birth would be ineffective in preventing spina bifida. 
         [0008]    In particular, neurologic development proceeds sequentially. The proper development of a structure depends on previous development of prior structures. Once a structure has developed incorrectly, the error is fixed and cannot later be revised. 
         [0009]    For example, the harmful effects of Down Syndrome on brain development begin before birth. A baby with Down Syndrome already has many characteristic features of the disease at the time of birth. There is often hypoplasis of the frontal lobes and cerebellum resulting in a reduced anteroposterior diameter of the head. This is commonly referred to as brachycephaly. The most important events in brain development are the earliest ones, and later developments are less important. As a result, there is a need for a method by which an abnormality, particularly a chromosomal abnormality, may be detected at any early stage of development, such as at the prenatal stage. A course of treatment may then be prescribed at a time that is beneficial to the baby. 
         [0010]    In abnormal fetal development, as in other pathologies, existing techniques can frequently identify a deficiency or abnormality in the existence or metabolism of a physiologically significant species. For example, various methods for analysis of target analytes generally, and evaluation of analytes in regards to prenatal testing specifically, have been proposed. Examples of some of these methods are disclosed in the following U.S. Pat. Nos.: 5,326,708, 5,438,017, 5,439,803, 5,506,150, 5,532,131, and 5,670,380. 
         [0011]    However, many vitamins are cofactors for multiple enzymes. Similarly, minerals can be cofactors for enzymes. A given metabolite might be a substrate in some reactions and/or a product in other reactions. A determination that a single metabolite, or a single vitamin or mineral is under or over-expressed in a particular pathology is often insufficient to provide a meaningful opportunity for therapeutic intervention. By comparison, comprehensive metabolic profiling would allow the physicians to see the whole panorama of metabolism and would allow an integrated, comprehensive characterization of the problem as well as an integrated, comprehensive approach to treatment. 
         [0012]    Accordingly, there is a need for a method to comprehensively profile the metabolic abnormalities found in the fetus with Down Syndrome or other chromosome abnormalities. Ideally, a global screen should be performed for a wide range of metabolites at one time. The results of the global screen could then be utilized to construct a biochemical profile that suggests treatment pathways for possible fetal chromosomal abnormalities including Down Syndrome. 
       SUMMARY OF THE INVENTION 
       [0013]    The present invention uses a sample of amniotic fluid to generate a comprehensive metabolic profile to diagnose chromosomal abnormalities in the fetus. The profile can be generated by several analytical techniques, and the components of the profile can be varied based on the clinical indication of interest. For example, a procedure similar to one described by Shoemaker and Elliott can be used to screen a specimen of amniotic fluid for metabolites. Shoemaker and Elliott,  Automated screening of urine samples for carbohydrates, organic and amino acids after treatment with urease , Journal of Chromatography, 562 (1991) 125-138, specifically incorporated herein by reference. A specimen of amniotic fluid is first obtained. For example, amniotic fluid may be taken from around the fetus during pregnancy. The specimen then is analyzed in a gas chromatograph/mass spectrophotometer (GC/MS). 
         [0014]    The results of the GC/MS analysis are then used to generate the profile of the metabolites previously identified. The sample profile is compared with a control profile of metabolites that is representative of the normal levels of those metabolites. By analyzing the sample profile with respect to the normal profile, each metabolite that has a different level when compared with the normal level of that metabolite can be identified. Using the identified metabolites that have different values than the norm, a biochemical treatment may be prescribed that addresses the concentration, i.e., to increase or decrease, of each of those metabolites or the metabolism or physiology thereof. Using this method, an improved treatment for chromosomal abnormalities such as Down Syndrome may be prescribed by taking into account metabolic deficiencies on a global level rather than individually or in small groups. 
     
    
     DETAILED DESCRIPTION OF THE INVENTION  
       [0015]    As previously discussed, a procedure similar to the procedure described by Shoemaker and Elliott may be used to screen an amniotic fluid specimen for metabolites. An amniotic fluid specimen is obtained from the fetus to be evaluated. For example, amniotic fluid may be obtained by placing a needle through the abdomen and uterine wall into the uterus and withdrawing the fluid with a syringe. It is preferable that the syringe not be lubricated with glycerol. The fluid may be separated from any unwanted cells by centrifugation. 
         [0016]    If storage of the specimen is required, the specimen is preferably maintained at −20° C. Subsequently, the specimen is transferred to a metabolic screening laboratory where a GC/MS apparatus is located. If the laboratory is remotely located, the specimen is shipped on dry ice. The specimen is then thawed in the laboratory in preparation for the test. 
         [0017]    The specimen is injected into the GC/MS apparatus in order to identify and measure the metabolites. All chemical constituents of the specimen are separated by their GC retention times. The identity and quantity of each chemical constituent is then determined by the MS. The MS preferably sweeps through all masses from 15-650 Daltons every two seconds. 
         [0018]    A comprehensive metabolic profile is then generated that indicates the identity and quantity of each metabolite. From these levels, the activity of the relevant enzymes related to each metabolite can be inferred. The metabolites included in the report may include, but are not limited to, organic acids, amino acids, glycine conjugates, fatty acids, vitamins, neurotransmitters, drugs, drug metabolites, hormones, and carbohydrates. 
         [0019]    The levels of the metabolites for the sample are then compared with the representative, normal level for each. The levels may be compared by examining mean levels and standard deviations for each metabolite. Alternatively, the levels may be compared using medians and a nonparametric analysis. By analyzing the relative levels of the metabolites as compared with the normal levels, a suitable biochemical treatment may be prescribed for a particular condition. 
         [0020]    Most of the metabolites are indicator substrates. An indicator substrate is a substrate that is metabolized by an enzyme. When the enzyme is deficient or its activity is low, the indicator substrate accumulates. When the enzyme activity is high, the metabolite may be low. If the activity of an enzyme is low, vitamin or mineral cofactors can be supplemented. If the activity of an enzyme is too high, it can be blocked or cofactors withheld. Some metabolites are vitamins or vitamin derivatives. If they are low, they can similarly be supplemented. 
         [0021]    Any metabolite that may be included in a GC/MS report or analogous analytical technique may be analyzed by the method of the present invention. This method is also used to analyze a global metabolic profile to provide the biochemical equivalent of a complete physical exam. The method detects abnormalities through the comprehensive metabolic profiling which identifies multiple metabolic abnormalities. Each one of these abnormalities, when present by itself in a severe form, would cause mental retardation. Abnormalities such as Down Syndrome may then be modeled as the sum of a group of simpler abnormalities that may have an identifiable metabolic profile. Many of these simpler abnormalities may have treatments that are suggested by the results of the comprehensive metabolic profiling. Using the results of the comprehensive metabolic profile, chromosomal abnormalities may be identified and proper treatment may be suggested at an opportune time. 
         [0022]    A key advantage of the method is the characterization of a disease state, a physiologic influence, or the effects of a drug. If a group of normal patients and a group of disease patients are characterized, the differences between the groups will form a biochemical characterization of the disease. 
       Example Profile Analysis 
       [0023]    To diagnose a fetus for chromosomal abnormalities using the method of the present invention, a metabolic profile must first be generated that is representative of the metabolite levels in an average patient suffering from the chromosomal abnormality that is to be diagnosed. Using the aforementioned GC/MS procedure, a metabolic profile for a group of 23 Down Syndrome patients was generated. The following tables represent the Down Syndrome metabolic profile separated into different metabolite groups. The tables also include results for a group of 41 normal patients, generated by the GC/MS procedure, for comparison purposes. Since the comparison data for a chromosomal abnormality may be analyzed by examining either the mean levels of the metabolites or the median levels of the metabolites, two tables are presented for each metabolite grouping with one table presenting mean levels and the other presenting median levels (i.e., a nonparametric analysis). Within each table, the Mann-Whitney p-value and the normal value v. abnormal value t-Test value are presented for each metabolite. Additionally, the standard deviation (S.D.) is presented for each metabolite in the tables that present the mean levels of the metabolites rather than the median levels. 
         [0024]    Table 1 illustrates a typical metabolic profile for the mean level of fatty acids within a population of Down Syndrome patients and a population of normal patients: 
         [0000]    
       
         
               
               
               
             
               
               
               
               
               
               
             
               
               
               
               
               
               
             
           
               
                 TABLE 1 
               
             
             
               
                   
               
               
                 Fatty Acid 
                 Normal 
                 Down Syndrome 
               
             
          
           
               
                 Compound 
                 t-Test 
                 Mean 
                 S.D. 
                 Mean 
                 S.D. 
               
               
                   
               
             
          
           
               
                 LAURIC ACID 
                 0.1874 
                 0.075 
                 0.2897 
                 0.3260 
                 0.8608 
               
               
                 MYRISTIC ACID 
                 0.5022 
                 2.2875 
                 3.0146 
                 3.0434 
                 4.8286 
               
               
                 PALMITOLEIC ACID 
                 0.8122 
                 0.0712 
                 0.2428 
                 0.0869 
                 0.2559 
               
               
                 PALMITIC ACID 
                 0.6203 
                 23.0625 
                 30.183 
                 18.7173 
                 34.919 
               
               
                 LINOLEIC ACID 
                 0.6124 
                 5.45 
                 7.1313 
                 7.7173 
                 20.4982 
               
               
                 LINOLENIC ACID 
                 0.4651 
                 3.3775 
                 5.8258 
                 5.4956 
                 12.9802 
               
               
                 OLEIC ACID 
                 0.6296 
                 39.8 
                 65.011 
                 30.4130 
                 78.4342 
               
               
                 STEARIC ACID 
                 0.6709 
                 3.1125 
                 3.6837 
                 2.6739 
                 4.0469 
               
               
                 ARACHIDONIC ACID 
                 0.7258 
                 0.5675 
                 2.3453 
                 0.4086 
                 1.2291 
               
               
                 EICOSAPENTAENOIC 
                 0.7013 
                 0.1237 
                 0.3415 
                 0.1 
                 0.1422 
               
               
                   
               
             
          
         
       
     
         [0025]    Table 2 illustrates a typical metabolic profile for the median level of fatty acids within a population of Down Syndrome patients and a population of normal patients: 
         [0000]    
       
         
               
               
               
               
             
               
               
               
               
             
           
               
                 TABLE 2 
               
               
                   
               
               
                 Fatty Acid 
                   
                 Normal 
                 Down Syndrome 
               
               
                 Compound 
                 p-value 
                 Median 
                 Median 
               
               
                   
               
             
             
               
                   
               
             
          
           
               
                 LAURIC ACID 
                 0.049 
                 0 
                 0 
               
               
                 MYRISTIC ACID 
                 0.575 
                 1.25 
                 1.5 
               
               
                 PALMITOLEIC ACID 
                 0.576 
                 0 
                 0 
               
               
                 PALMITIC ACID 
                 0.342 
                 11.75 
                 10.5 
               
               
                 LINOLEIC ACID 
                 1 
                 2.25 
                 2.5 
               
               
                 LINOLENIC ACID 
                 0.91 
                 0.675 
                 0.8 
               
               
                 OLEIC ACID 
                 0.391 
                 11.5 
                 10 
               
               
                 STEARIC ACID 
                 0.564 
                 1.5 
                 1.5 
               
               
                 ARACHIDONIC ACID 
                 0.103 
                 0 
                 0 
               
               
                 EICOSAPENTAENOIC 
                 0.361 
                 0 
                 0 
               
               
                   
               
             
          
         
       
     
         [0026]    Table 3 illustrates a typical metabolic profile for the mean level of simple sugars within a population of Down Syndrome patients and a population of normal patients: 
         [0000]    
       
         
               
               
               
             
               
               
               
               
               
               
             
               
               
               
               
               
               
             
           
               
                 TABLE 3 
               
             
             
               
                   
               
               
                 Simple Sugars 
                 Normal 
                 Down Syndrome 
               
             
          
           
               
                 Compound 
                 t-Test 
                 Mean 
                 S.D. 
                 Mean 
                 S.D. 
               
               
                   
               
             
          
           
               
                 THREITOL 
                 0.7620 
                 7.8875 
                 4.6979 
                 7.5217 
                 4.5263 
               
               
                 ERYTHRITOL 
                 0.1094 
                 6.9875 
                 5.3594 
                 4.5869 
                 5.7537 
               
               
                 ARABINOSE 
                 0.5616 
                 0.6375 
                 0.6503 
                 0.7391 
                 0.6719 
               
               
                 FUCOSE 
                 0.2285 
                 0.3975 
                 1.1847 
                 0.1630 
                 0.2001 
               
               
                 RIBOSE 
                 0.3449 
                 0.6425 
                 0.3888 
                 0.7652 
                 0.5394 
               
               
                 ERYTHRITOL.2 
                 0.2743 
                 9.3625 
                 5.3011 
                 11.1956 
                 6.8286 
               
               
                 FRUCTOSE 
                 0.5252 
                 27.1 
                 43.9131 
                 17.7391 
                 61.5203 
               
               
                 GLUCOSE mg/dL 
                 0.7164 
                 21.3 
                 24.4613 
                 19.1739 
                 20.8744 
               
               
                 GALACTOSE 
                 0.5750 
                 1856.16 
                 2477.33 
                 1545.69 
                 1856.03 
               
               
                 MANNOSE 
                 0.2175 
                 3.7625 
                 3.2874 
                 2.8913 
                 2.2409 
               
               
                 N-AC-GLUCOSAMINE 
                 0.5290 
                 0.3225 
                 0.4437 
                 0.2652 
                 0.2740 
               
               
                 LACTOSE 
                 0.4813 
                 0.625 
                 0.9919 
                 0.8043 
                 0.9503 
               
               
                 MALTOSE 
                 0.3696 
                 0.6125 
                 1.0830 
                 0.8913 
                 1.2243 
               
               
                 XYLITOL 
                 0.7618 
                 6.465 
                 8.2362 
                 5.9804 
                 4.3894 
               
               
                 ARABINITOL 
                 0.1907 
                 0.4862 
                 1.5326 
                 0.1586 
                 0.2097 
               
               
                 RIBITOL 
                 0.9760 
                 0.195 
                 0.4903 
                 0.1978 
                 0.2524 
               
               
                 ALLOSE 
                 0.3697 
                 0.7512 
                 1.7308 
                 0.4869 
                 0.4873 
               
               
                 GLUCURONIC ACID 
                 0.3784 
                 59.7912 
                 137.818 
                 36.8978 
                 66.2477 
               
               
                 GALACTONIC ACID 
                 0.9373 
                 149.55 
                 776.895 
                 162.347 
                 507.218 
               
               
                 GLUCONIC ACID 
                 0.8097 
                 0.3587 
                 0.7182 
                 0.3913 
                 0.3466 
               
               
                 GLUCARIC 
                 0.3784 
                 0.06 
                 0.0968 
                 0.0913 
                 0.1512 
               
               
                 MANNITOL 
                 0.8416 
                 45.9612 
                 120.018 
                 50.9195 
                 75.9496 
               
               
                 DULCITOL 
                 0.4140 
                 0.1312 
                 0.6863 
                 0.0413 
                 0.0468 
               
               
                 SORBITOL 
                 0.3269 
                 0.15 
                 0.1870 
                 0.2086 
                 0.2452 
               
               
                 INOSITOL 
                 0.9174 
                 26.8425 
                 26.0067 
                 27.4782 
                 21.6581 
               
               
                 SUCROSE 
                 n/a 
                 0 
                 0 
                 0 
                 0 
               
               
                 METANEPHRINE mg/DL 
                 0.3281 
                 0 
                 0 
                 0.0021 
                 0.0104 
               
               
                   
               
             
          
         
       
     
         [0027]    Table 4 illustrates a typical metabolic profile for the median level of simple sugars within a population of Down Syndrome patients and a population of normal patients: 
         [0000]    
       
         
               
               
               
               
             
               
               
               
               
             
           
               
                 TABLE 4 
               
               
                   
               
               
                 Simple Sugars 
                   
                 Normal 
                 Down Syndrome 
               
               
                 Compound 
                 p-value 
                 Median 
                 Median 
               
               
                   
               
             
             
               
                   
               
             
          
           
               
                 THREITOL 
                 0.627 
                 7.25 
                 6.5 
               
               
                 ERYTHRITOL 
                 0.057 
                 7 
                 1.5 
               
               
                 ARABINOSE 
                 0.504 
                 0.5 
                 0.5 
               
               
                 FUCOSE 
                 0.192 
                 0.05 
                 0.1 
               
               
                 RIBOSE 
                 0.663 
                 0.675 
                 0.65 
               
               
                 ERYTHRITOL.2 
                 0.431 
                 8 
                 8.5 
               
               
                 FRUCTOSE 
                 0.102 
                 1.5 
                 1.5 
               
               
                 GLUCOSE mg/dL 
                 0.716 
                 12.5 
                 11.5 
               
               
                 GALACTOSE 
                 0.498 
                 1056.5 
                 692 
               
               
                 MANNOSE 
                 0.352 
                 3 
                 3 
               
               
                 N-AC-GLUCOSAMINE 
                 0.317 
                 0.1 
                 0.2 
               
               
                 LACTOSE 
                 0.204 
                 0.5 
                 0.5 
               
               
                 MALTOSE 
                 0.231 
                 0 
                 0.5 
               
               
                 XYLITOL 
                 0.602 
                 4.3 
                 5.05 
               
               
                 ARABINITOL 
                 0.455 
                 0.1 
                 0.1 
               
               
                 RIBITOL 
                 0.099 
                 0 
                 0.1 
               
               
                 ALLOSE 
                 0.769 
                 0.3 
                 0.35 
               
               
                 GLUCURONIC ACID 
                 0.797 
                 4.65 
                 1.8 
               
               
                 GALACTONIC ACID 
                 0.853 
                 8 
                 9 
               
               
                 GLUCONIC ACID 
                 0.085 
                 0.175 
                 0.35 
               
               
                 GLUCARIC 
                 0.471 
                 0 
                 0.05 
               
               
                 MANNITOL 
                 0.264 
                 0.325 
                 0.7 
               
               
                 DULCITOL 
                 0.053 
                 0 
                 0.05 
               
               
                 SORBITOL 
                 0.336 
                 0.1 
                 0.15 
               
               
                 INOSITOL 
                 0.462 
                 16.75 
                 27.5 
               
               
                 SUCROSE 
                 1 
                 0 
                 0 
               
               
                 METANEPHRINE mg/dL 
                 0.187 
                 0 
                 0 
               
               
                   
               
             
          
         
       
     
         [0028]    Table 5 illustrates a typical metabolic profile for the mean level of amino acids and glycine conjugates within a population of Down Syndrome patients and a population of normal patients: 
         [0000]    
       
         
               
               
               
             
               
               
               
               
               
               
             
               
               
               
               
               
               
             
           
               
                 TABLE 5 
               
             
             
               
                   
               
               
                 Amino Acids and 
                   
                   
               
               
                 Glycine Conjugates 
                 Normal 
                 Down Syndrome 
               
             
          
           
               
                 Compound 
                 t-Test 
                 Mean 
                 S.D. 
                 Mean 
                 S.D. 
               
               
                   
               
             
          
           
               
                 PROPIONYL GLY 
                 0.9948 
                 0.2225 
                 0.6289 
                 0.2217 
                 0.2848 
               
               
                 BUTYRYL GLYCINE 
                 0.8212 
                 0.2213 
                 0.6913 
                 0.2565 
                 0.5292 
               
               
                 HEXANOYL GLYCINE 
                 0.4152 
                 0.3938 
                 1.3668 
                 0.2087 
                 0.3006 
               
               
                 PHENYL PROP GLY 
                 0.7414 
                 0.5000 
                 2.3993 
                 0.3543 
                 1.0615 
               
               
                 SUBERYL GLYCINE 
                 0.5090 
                 0.0300 
                 0.0890 
                 0.0457 
                 0.0903 
               
               
                 ISOVALERYL GLY 
                 0.9786 
                 0.4650 
                 1.9601 
                 0.4543 
                 1.1773 
               
               
                 TIGLY GLY 
                 0.4076 
                 0.3625 
                 0.7605 
                 0.2457 
                 0.3447 
               
               
                 BETA MET CROT GLY 
                 0.4827 
                 1.2750 
                 4.3378 
                 0.7696 
                 0.9522 
               
               
                 GLYCINE 
                 0.3641 
                 276.2250 
                 199.0262 
                 229.2826 
                 193.7927 
               
               
                 ALANINE 
                 0.0337 
                 455.3000 
                 491.5694 
                 202.5870 
                 412.2851 
               
               
                 SARCOSINE 
                 0.2933 
                 1.0988 
                 1.2466 
                 0.7826 
                 1.0703 
               
               
                 BETA-ALANINE 
                 0.9038 
                 17.2700 
                 11.8828 
                 16.9109 
                 10.9422 
               
               
                 B-AMINOISOBUTYRIC 
                 0.2457 
                 2.2538 
                 2.7931 
                 1.6609 
                 1.1794 
               
               
                 SERINE 
                 0.2684 
                 19.7875 
                 17.7825 
                 15.1087 
                 14.8561 
               
               
                 PROLINE 
                 0.0385 
                 318.4875 
                 322.0118 
                 186.5435 
                 172.6874 
               
               
                 HYDROXY PROLINE 
                 0.1647 
                 81.1000 
                 50.1975 
                 105.9565 
                 74.7918 
               
               
                 HYDROXY LYSINE 
                 0.9268 
                 0.7413 
                 1.2271 
                 0.7152 
                 0.9819 
               
               
                 ASPARTIC ACID 
                 0.1032 
                 6.7913 
                 5.0833 
                 11.1761 
                 11.8518 
               
               
                 ASPARAGINE 
                 0.7315 
                 0.6338 
                 0.7522 
                 0.6913 
                 0.5618 
               
               
                 N-AC ASPARTIC 
                 0.8949 
                 0.1800 
                 0.6900 
                 0.1978 
                 0.3761 
               
               
                 ORNITHINE 
                 0.3084 
                 40.8000 
                 29.5179 
                 32.6522 
                 30.6162 
               
               
                 GLUTAMIC ACID 
                 0.9576 
                 261.4500 
                 204.4625 
                 258.6522 
                 197.4870 
               
               
                 GLUTAMINE 
                 0.1047 
                 1.0250 
                 3.0633 
                 3.5870 
                 6.9407 
               
               
                 PIPECOLIC ACID 
                 0.7858 
                 3.4488 
                 2.7644 
                 3.7065 
                 3.9971 
               
               
                 LEUCINE 
                 0.0073 
                 231.5000 
                 181.3793 
                 126.5652 
                 117.7761 
               
               
                 KETO LEUCINE 
                 0.8192 
                 53.0900 
                 39.8797 
                 50.0196 
                 56.3368 
               
               
                 VALINE 
                 0.0440 
                 383.8125 
                 302.1702 
                 239.1087 
                 246.4417 
               
               
                 KETO-VALINE 
                 0.9095 
                 13.8475 
                 12.2042 
                 14.2370 
                 13.4663 
               
               
                 ISOLEUCINE 
                 0.0651 
                 103.6875 
                 86.9055 
                 67.2609 
                 65.4288 
               
               
                 KETO-ISOLEUCINE 
                 0.7580 
                 7.5625 
                 7.5049 
                 8.2000 
                 8.0510 
               
               
                 LYSINE 
                 0.7142 
                 366.5500 
                 287.9107 
                 341.2174 
                 247.3257 
               
               
                 HISTIDINE 
                 0.8280 
                 14.7000 
                 12.2138 
                 14.0217 
                 11.6643 
               
               
                 THREONINE 
                 0.2161 
                 292.3125 
                 277.0502 
                 217.4348 
                 195.7553 
               
               
                 HOMOSERINE 
                 0.5177 
                 0.3688 
                 0.3549 
                 0.4261 
                 0.3250 
               
               
                 METHIONINE 
                 0.5314 
                 34.8375 
                 30.2578 
                 40.2609 
                 34.2282 
               
               
                 CYSTEINE 
                 0.1124 
                 152.9625 
                 103.4822 
                 212.1957 
                 155.5338 
               
               
                 HOMOCYSTEINE 
                 0.0690 
                 0.0625 
                 0.1436 
                 0.1565 
                 0.2139 
               
               
                 CYSTATHIONINE 
                 0.2705 
                 0.1288 
                 0.2287 
                 0.2087 
                 0.2957 
               
               
                 HOMOCYSTINE 
                 0.7456 
                 0.0688 
                 0.1846 
                 0.0565 
                 0.1131 
               
               
                 CYSTINE 
                 0.6096 
                 0.3138 
                 0.7375 
                 0.2457 
                 0.3030 
               
               
                 PHENYLALANINE 
                 0.2054 
                 144.6500 
                 79.8540 
                 180.2174 
                 117.4317 
               
               
                 TYROSINE 
                 0.7914 
                 107.1750 
                 98.7430 
                 100.6087 
                 91.7593 
               
               
                 TRYPTOPHAN 
                 0.9187 
                 10.3875 
                 10.1908 
                 10.1087 
                 10.4870 
               
               
                   
               
             
          
         
       
     
         [0029]    Table 6 illustrates a typical metabolic profile for the median level of amino acids and glycine conjugates within a population of Down Syndrome patients and a population of normal patients: 
         [0000]    
       
         
               
               
               
               
             
               
               
               
               
             
           
               
                 TABLE 6 
               
               
                   
               
               
                 Amino Acids and 
                   
                   
                   
               
               
                 Glycine Conjugates 
                   
                 Normal 
                 Down Syndrome 
               
               
                 Compound 
                 p-value 
                 Median 
                 Median 
               
               
                   
               
             
             
               
                   
               
             
          
           
               
                 PROPIONYL GLY 
                 0.108 
                 0 
                 0.15 
               
               
                 BUTYRYL GLYCINE 
                 0.02 
                 0 
                 0.1 
               
               
                 HEXANOYL GLYCINE 
                 0.011 
                 0 
                 0.1 
               
               
                 PHENYL PROP GLY 
                 0.933 
                 0 
                 0 
               
               
                 SUBERYL GLYCINE 
                 0.148 
                 0 
                 0 
               
               
                 ISOVALERYL GLY 
                 0.005 
                 0 
                 0.1 
               
               
                 TIGLY GLY 
                 0.976 
                 0.125 
                 0.1 
               
               
                 BETA MET CROT GLY 
                 0.41 
                 0.175 
                 0.55 
               
               
                 GLYCINE 
                 0.357 
                 280 
                 249.5 
               
               
                 ALANINE 
                 0.069 
                 312.25 
                 2 
               
               
                 SARCOSINE 
                 0.399 
                 0.7 
                 0.25 
               
               
                 BETA-ALANINE 
                 1 
                 16.025 
                 18.25 
               
               
                 B-AMINOISOBUTYRIC 
                 0.937 
                 1.35 
                 1.4 
               
               
                 SERINE 
                 0.247 
                 17.25 
                 13 
               
               
                 PROLINE 
                 0.265 
                 199.5 
                 129 
               
               
                 HYDROXY PROLINE 
                 0.214 
                 68.5 
                 86.5 
               
               
                 HYDROXY LYSINE 
                 0.552 
                 0.225 
                 0.3 
               
               
                 ASPARTIC ACID 
                 0.454 
                 6.35 
                 8 
               
               
                 ASPARAGINE 
                 0.399 
                 0.475 
                 0.45 
               
               
                 N-AC ASPARTIC 
                 0.006 
                 0 
                 0.05 
               
               
                 ORNITHINE 
                 0.138 
                 34.25 
                 24 
               
               
                 GLUTAMIC ACID 
                 0.926 
                 212.75 
                 249 
               
               
                 GLUTAMINE 
                 0.002 
                 0.5 
                 1 
               
               
                 PIPECOLIC ACID 
                 0.903 
                 2.8 
                 2.75 
               
               
                 LEUCINE 
                 0.036 
                 221.25 
                 93.5 
               
               
                 KETO LEUCINE 
                 0.457 
                 49.725 
                 36.3 
               
               
                 VALINE 
                 0.085 
                 368.5 
                 176.5 
               
               
                 KETO-VALINE 
                 0.972 
                 11 
                 9.9 
               
               
                 ISOLEUCINE 
                 0.123 
                 93.5 
                 51 
               
               
                 KETO-ISOLEUCINE 
                 0.689 
                 6.125 
                 6.25 
               
               
                 LYSINE 
                 0.627 
                 280.25 
                 270 
               
               
                 HISTIDINE 
                 0.732 
                 11.5 
                 11 
               
               
                 THREONINE 
                 0.265 
                 236.75 
                 155 
               
               
                 HOMOSERINE 
                 0.405 
                 0.3 
                 0.4 
               
               
                 METHIONINE 
                 0.638 
                 33.35 
                 38.7 
               
               
                 CYSTEINE 
                 0.104 
                 144 
                 200 
               
               
                 HOMOCYSTEINE 
                 0.01 
                 0 
                 0.1 
               
               
                 CYSTATHIONINE 
                 0.09 
                 0.05 
                 0.1 
               
               
                 HOMOCYSTINE 
                 0.114 
                 0 
                 0 
               
               
                 CYSTINE 
                 0.115 
                 0.075 
                 0.15 
               
               
                 PHENYLALANINE 
                 0.467 
                 138 
                 150.5 
               
               
                 TYROSINE 
                 0.663 
                 66 
                 67 
               
               
                 TRYPTOPHAN 
                 0.869 
                 6.25 
                 4.5 
               
               
                   
               
             
          
         
       
     
         [0030]    Table 7 illustrates a typical metabolic profile for the mean level of neurotransmitters within a population of Down Syndrome patients and a population of normal patients: 
         [0000]    
       
         
               
               
               
             
               
               
               
               
               
               
             
               
               
               
               
               
               
             
           
               
                 TABLE 7 
               
             
             
               
                   
               
               
                 Neurotransmitters 
                 Normal 
                 Down Syndrome 
               
             
          
           
               
                 Compound 
                 t-Test 
                 Mean 
                 S.D. 
                 Mean 
                 S.D. 
               
               
                   
               
             
          
           
               
                 GABA 
                 0.9990 
                 0.8063 
                 0.7186 
                 0.8065 
                 0.8286 
               
               
                 HOMOVANILLIC ACID 
                 0.5579 
                 0.1950 
                 0.3004 
                 0.2609 
                 0.4824 
               
               
                 NORMETANEPHRINE 
                 0.0388 
                 0.0013 
                 0.0079 
                 0.0391 
                 0.0825 
               
               
                 VANILLYLMANDELIC 
                 0.0849 
                 0.0038 
                 0.0175 
                 0.0152 
                 0.0279 
               
               
                 METANEPHRINE 
                 0.5954 
                 0.6488 
                 2.2671 
                 0.4022 
                 1.3971 
               
               
                 5-HIAA 
                 0.5821 
                 0.0100 
                 0.0258 
                 0.0065 
                 0.0229 
               
               
                 MHPG 
                 0.5438 
                 0.0038 
                 0.0175 
                 0.0065 
                 0.0172 
               
               
                 ETHANOLAMINE 
                 0.6039 
                 63.3375 
                 46.5176 
                 56.7826 
                 48.7374 
               
               
                   
               
             
          
         
       
     
         [0031]    Table 8 illustrates a typical metabolic profile for the median level of neurotransmitters within a population of Down Syndrome patients and a population of normal patients: 
         [0000]    
       
         
               
               
               
               
             
               
               
               
               
             
           
               
                 TABLE 8 
               
               
                   
               
               
                 Neurotransmitters 
                   
                 Normal 
                 Down Syndrome 
               
               
                 Compound 
                 p-value 
                 Median 
                 Median 
               
               
                   
               
             
             
               
                   
               
             
          
           
               
                 GABA 
                 0.539 
                 0.75 
                 0.45 
               
               
                 HOMOVANILLIC ACID 
                 0.663 
                 0.1 
                 0.15 
               
               
                 NORMETANEPHRINE 
                 0.001 
                 0 
                 0 
               
               
                 VANILLYLMANDELIC 
                 0.019 
                 0 
                 0 
               
               
                 METANEPHRINE 
                 0.707 
                 0.075 
                 0.1 
               
               
                 5-HIAA 
                 0.489 
                 0 
                 0 
               
               
                 MHPG 
                 0.279 
                 0 
                 0 
               
               
                 ETHANOLAMINE 
                 0.753 
                 65.5 
                 65 
               
               
                   
               
             
          
         
       
     
         [0032]    Table 9 illustrates a typical metabolic profile for the mean level of nutritionals within a population of Down Syndrome patients and a population of normal patients: 
         [0000]    
       
         
               
               
               
             
               
               
               
               
               
               
             
               
               
               
               
               
               
             
           
               
                 TABLE 9 
               
             
             
               
                   
               
               
                 Nutritionals 
                 Normal 
                 Down Syndrome 
               
             
          
           
               
                 Compound 
                 t-Test 
                 Mean 
                 S.D. 
                 Mean 
                 S.D. 
               
               
                   
               
             
          
           
               
                 FORMIMINOGLUTAMIC ACID 
                 0.0033 
                 1.4877 
                 0.0462 
                 0.8829 
                 0.7441 
               
               
                 4-PYRIDOXIC ACID 
                 0.4924 
                 1.2863 
                 1.8158 
                 1.0370 
                 1.0489 
               
               
                 PANTOTHENIC ACID 
                 0.8733 
                 299.3500 
                 527.7872 
                 278.8043 
                 466.6548 
               
               
                 XANTHURENIC ACID 407 
                 0.3163 
                 0.0400 
                 0.2214 
                 0.0043 
                 0.0144 
               
               
                 KYNURENINE 
                 0.8218 
                 1.2075 
                 1.6656 
                 1.1283 
                 1.1077 
               
               
                 QUINOLINIC 
                 0.8759 
                 0.1913 
                 1.0503 
                 0.2261 
                 0.7077 
               
               
                 OROTIC ACID 
                 0.1702 
                 0.0133 
                 0.0502 
                 0.0543 
                 0.1344 
               
               
                 D-AM LEVULINIC 
                 0.1741 
                 11.3150 
                 23.8991 
                 5.8217 
                 6.0014 
               
               
                 3-METHYL HISTIDINE 
                 0.8620 
                 1.6000 
                 1.9553 
                 1.5217 
                 1.5556 
               
               
                 NIACINAMIDE 
                 0.8650 
                 0.8825 
                 4.2032 
                 0.7587 
                 1.3854 
               
               
                 PSEUDOURIDINE 
                 0.7538 
                 0.2875 
                 0.4220 
                 0.3261 
                 0.4910 
               
               
                 2-DEOXYTETRONIC 
                 0.2681 
                 1.2250 
                 1.1033 
                 1.5435 
                 1.0757 
               
               
                 P-HO-PHEN-ACETIC 
                 0.5254 
                 0.1125 
                 0.2399 
                 0.1522 
                 0.2352 
               
               
                 XANTHINE 
                 0.6903 
                 0.0488 
                 0.1508 
                 0.0630 
                 0.1272 
               
               
                 UROCANIC ACID 
                 0.9655 
                 0.0625 
                 0.3240 
                 0.0652 
                 0.1722 
               
               
                 ASCORBIC ACID 
                 0.3415 
                 0.1000 
                 0.4961 
                 0.0217 
                 0.1043 
               
               
                 GLYCEROL 
                 0.4712 
                 36.6663 
                 28.3624 
                 31.4870 
                 26.5956 
               
               
                   
               
             
          
         
       
     
         [0033]    Table 10 illustrates a typical metabolic profile for the median level of nutritionals within a population of Down Syndrome patients and a population of normal patients: 
         [0000]    
       
         
               
               
               
               
             
               
               
               
               
             
           
               
                 TABLE 10 
               
               
                   
               
               
                 Nutritionals 
                   
                 Normal 
                 Down Syndrome 
               
               
                 Compound 
                 p-value 
                 Median 
                 Median 
               
               
                   
               
             
             
               
                   
               
             
          
           
               
                 FORMIMINOGLUTAMIC 
                 0.007 
                 1.3425 
                 0.9119 
               
               
                 ACID 
               
               
                 4-PYRIDOXIC ACID 
                 0.839 
                 0.8 
                 0.8 
               
               
                 PANTOTHENIC ACID 
                 0.932 
                 149.5 
                 151.5 
               
               
                 XANTHURENIC ACID 407 
                 0.822 
                 0 
                 0 
               
               
                 KYNURENINE 
                 0.303 
                 0.425 
                 0.8 
               
               
                 QUINOLINIC 
                 0.199 
                 0 
                 0 
               
               
                 OROTIC ACID 
                 0.189 
                 0 
                 0 
               
               
                 D-AM LEVULINIC 
                 0.587 
                 2.85 
                 2.6 
               
               
                 3-METHYL HISTIDINE 
                 0.712 
                 1 
                 1 
               
               
                 NIACINAMIDE 
                 0.044 
                 0 
                 0.25 
               
               
                 PSEUDOURIDINE 
                 0.775 
                 0 
                 0 
               
               
                 2-DEOXYTETRONIC 
                 0.214 
                 1 
                 1.5 
               
               
                 P-HO-PHEN-ACETIC 
                 0.39 
                 0 
                 0 
               
               
                 XANTHINE 
                 0.653 
                 0 
                 0 
               
               
                 UROCANIC ACID 
                 0.279 
                 0 
                 0 
               
               
                 ASCORBIC ACID 
                 0.877 
                 0 
                 0 
               
               
                 GLYCEROL 
                 0.539 
                 35.525 
                 31.85 
               
               
                   
               
             
          
         
       
     
         [0034]    Table 11 illustrates a typical metabolic profile for the mean level of organic acids within a population of Down Syndrome patients and a population of normal patients: 
         [0000]    
       
         
               
               
               
             
               
               
               
               
               
               
             
               
               
               
               
               
               
             
           
               
                 TABLE 11 
               
             
             
               
                   
               
               
                 Organic Acids 
                 Normal 
                 Down Syndrome 
               
             
          
           
               
                 Compound 
                 t-Test 
                 Mean 
                 S.D. 
                 Mean 
                 S.D. 
               
               
                   
               
             
          
           
               
                 LACTIC ACID uM/L 
                 0.3580 
                 7455.3375 
                 6476.7984 
                 6096.8478 
                 5027.6119 
               
               
                 PYRUVIC ACID 
                 0.0563 
                 16.1375 
                 22.5374 
                 7.9783 
                 10.5760 
               
               
                 GLYCOLIC ACID 
                 0.3684 
                 25.2375 
                 65.5558 
                 43.5435 
                 82.6529 
               
               
                 ALPHA-OH-BUTYRIC 
                 0.0965 
                 21.4750 
                 22.4214 
                 11.6304 
                 22.0337 
               
               
                 OXALIC ACID 
                 0.0239 
                 12.7000 
                 21.4652 
                 34.8913 
                 41.5667 
               
               
                 4-OH-BUTYRIC 
                 0.4037 
                 0.1000 
                 0.4114 
                 0.1957 
                 0.4457 
               
               
                 HEXANOIC ACID 
                 0.9894 
                 11.1375 
                 21.0619 
                 11.0652 
                 20.5234 
               
               
                 5-HYDROXYCAPROIC 
                 0.5482 
                 0.2100 
                 0.9129 
                 0.3239 
                 0.5825 
               
               
                 OCTANOIC 
                 0.9308 
                 0.6625 
                 1.8271 
                 0.6304 
                 1.0894 
               
               
                 BETA-LACTATE 
                 0.0313 
                 11.9625 
                 13.0852 
                 6.1522 
                 7.8327 
               
               
                 SUCCINIC ACID 
                 0.7615 
                 1.2125 
                 1.5603 
                 1.3261 
                 1.3366 
               
               
                 GLUTARIC ACID 
                 0.3675 
                 0.5588 
                 2.2167 
                 0.2261 
                 0.4970 
               
               
                 2-OXO-GLUTARATE 
                 0.1486 
                 6.1250 
                 13.4273 
                 11.9130 
                 15.8592 
               
               
                 FUMARIC 
                 0.8900 
                 0.1275 
                 0.3629 
                 0.1174 
                 0.2146 
               
               
                 MALEIC 
                 0.0986 
                 2.7875 
                 3.1928 
                 4.7348 
                 4.9316 
               
               
                 MALIC ACID 
                 0.4096 
                 2.0000 
                 2.7087 
                 2.5587 
                 2.4817 
               
               
                 ADIPIC ACID 
                 0.8252 
                 0.1375 
                 0.5126 
                 0.1174 
                 0.1940 
               
               
                 SUBERIC ACID 
                 0.7434 
                 0.1925 
                 0.3964 
                 0.2196 
                 0.2557 
               
               
                 SEBACIC ACID 
                 0.0816 
                 0.6875 
                 0.8276 
                 0.3413 
                 0.6933 
               
               
                 GLYCERIC ACID 
                 0.6070 
                 28.0000 
                 22.2955 
                 25.2609 
                 18.9406 
               
               
                 BETA-OH-BUTYRIC 
                 0.4624 
                 61.4750 
                 53.3212 
                 50.2174 
                 60.5152 
               
               
                 METHYLSUCCINIC 
                 0.1623 
                 0.1775 
                 0.7137 
                 0.5087 
                 0.9728 
               
               
                 METHYLMALONIC 
                 0.9946 
                 25.3988 
                 23.5841 
                 25.4348 
                 18.3025 
               
               
                 ETHYLMALONIC 
                 0.3967 
                 28.3250 
                 65.1132 
                 16.7174 
                 42.6016 
               
               
                 HOMOGENTISIC ACID 
                 0.3281 
                 0.0138 
                 0.0519 
                 0.0370 
                 0.1047 
               
               
                 PHENYLPYRUVIC ACID 
                 0.0254 
                 0.0375 
                 0.0749 
                 0.1435 
                 0.2063 
               
               
                 SUCCINYLACETONE 
                 0.9787 
                 0.2600 
                 0.5466 
                 0.2630 
                 0.3549 
               
               
                 3-OH-ISOVALERIC 
                 0.5760 
                 3.5813 
                 3.9367 
                 3.0652 
                 3.2301 
               
               
                 PHOSPHATE mg/dL 
                 0.6839 
                 1.0363 
                 1.2176 
                 0.9174 
                 1.0422 
               
               
                 CITRIC ACID 
                 0.9542 
                 36.7000 
                 47.5228 
                 37.3261 
                 37.4884 
               
               
                 HIPPURIC ACID 
                 0.0613 
                 21.4125 
                 24.8725 
                 12.9783 
                 9.6218 
               
               
                 URIC ACID mg/dL 
                 0.5409 
                 0.4200 
                 0.7250 
                 0.3391 
                 0.3086 
               
               
                   
               
             
          
         
       
     
         [0035]    Table 12 illustrates a typical metabolic profile for the median level of organic acids within a population of Down Syndrome patients and a population of normal patients: 
         [0000]    
       
         
               
               
               
               
             
               
               
               
               
             
           
               
                 TABLE 12 
               
               
                   
               
               
                 Organic Acids 
                   
                 Normal 
                 Down Syndrome 
               
               
                 Compound 
                 p-value 
                 Median 
                 Median 
               
               
                   
               
             
             
               
                   
               
             
          
           
               
                 LACTIC ACID uM/L 
                 0.521 
                 5563.5 
                 5138 
               
               
                 PYRUVIC ACID 
                 0.311 
                 5.25 
                 5 
               
               
                 GLYCOLIC ACID 
                 0.668 
                 12.5 
                 13 
               
               
                 ALPHA-OH-BUTYRIC 
                 0.046 
                 17.25 
                 1 
               
               
                 OXALIC ACID 
                 0.007 
                 5 
                 23.5 
               
               
                 4-OH-BUTYRIC 
                 0.103 
                 0 
                 0 
               
               
                 HEXANOIC ACID 
                 0.731 
                 6.25 
                 4 
               
               
                 5-HYDROXYCAPROIC 
                 0.009 
                 0 
                 0.05 
               
               
                 OCTANOIC 
                 0.643 
                 0 
                 0 
               
               
                 BETA-LACTATE 
                 0.198 
                 6 
                 4.5 
               
               
                 SUCCINIC ACID 
                 0.433 
                 0.5 
                 1 
               
               
                 GLUTARIC ACID 
                 0.095 
                 0 
                 0.05 
               
               
                 2-OXO-GLUTARATE 
                 0.034 
                 1 
                 5 
               
               
                 FUMARIC 
                 0.175 
                 0 
                 0.05 
               
               
                 MALEIC 
                 0.108 
                 1.525 
                 2.5 
               
               
                 MALIC ACID 
                 0.252 
                 0.975 
                 1.6 
               
               
                 ADIPIC ACID 
                 0.009 
                 0 
                 0.05 
               
               
                 SUBERIC ACID 
                 0.162 
                 0.05 
                 0.1 
               
               
                 SEBACIC ACID 
                 0.13 
                 0.35 
                 0.1 
               
               
                 GLYCERIC ACID 
                 0.814 
                 25.25 
                 26 
               
               
                 BETA-OH-BUTYRIC 
                 0.414 
                 66.25 
                 33 
               
               
                 METHYLSUCCINIC 
                 0.002 
                 0 
                 0.15 
               
               
                 METHYLMALONIC 
                 0.563 
                 21.3 
                 24.3 
               
               
                 ETHYLMALONIC 
                 0.407 
                 6.125 
                 2.8 
               
               
                 HOMOGENTISIC ACID 
                 0.218 
                 0 
                 0 
               
               
                 PHENYLPYRUVIC ACID 
                 0.017 
                 0 
                 0.05 
               
               
                 SUCCINYLACETONE 
                 0.188 
                 0.05 
                 0.1 
               
               
                 3-OH-ISOVALERIC 
                 0.881 
                 2.375 
                 1.45 
               
               
                 PHOSPHATE mg/dL 
                 0.949 
                 0.65 
                 0.7 
               
               
                 CITRIC ACID 
                 0.38 
                 13 
                 22.5 
               
               
                 HIPPURIC ACID 
                 0.287 
                 11.25 
                 9 
               
               
                 URIC ACID mg/dL 
                 0.426 
                 0.2 
                 0.25 
               
               
                   
               
             
          
         
       
     
         [0036]    If a metabolic profile of a specific patient reflected the same profile as the Down Syndrome group, the method of the present invention could be used to diagnose and suggest treatment for this patient. In the example profile data, formiminoglutamate (FIGLU) in Down Syndrome (Mean=0.8829, Median=0.9119) is decreased from normal (Mean=1.4877, Median=1.3425). (See Tables 9 and 10). FIGLU is a metabolite of histidine, which contributes a mono-carbon to tetrahydrofolate. In folate deficiency, FIGLU accumulates because there is little folate to accept the mono-carbon. Therefore, folate deficiency results in a shortage of mono-carbon tetrahydrofolate. Additionally, mono-carbons are necessary to synthesize biologically important molecules. Mono-carbon tetrahydrofolate is a major source of mono-carbons used in cellular biosynthesis. Mono-carbons are required to synthesize purines (e.g. adenine &amp; guanine), components of DNA, RNA, and other important molecules. In Down Syndrome, purine synthesis is accelerated. There are signs of mono-carbon shortage but folate is not deficient. In our data, we see that FIGLU is reduced. This probably reflects an increased demand for mono-carbons due to accelerated purine synthesis. Therefore, a treatment of mono-carbon supplements could be prescribed for this patient. 
         [0037]    Another metabolite that should be addressed according to this data is normetanephrine. (See Tables 7 and 8). Normetanephrine is a metabolite of the neurotransmitter norepinephrine (NE). NE requires a mono-carbon to be metabolized to epinephrine. Mono-carbon shortage explains the finding of elevated normetanephrine in Down Syndrome (Mean=0.039, Median=0) compared to normal amniotic fluid (Mean=0.0013, Median=0). 
         [0038]    Cystathionine beta synthase (CBS) is an enzyme whose gene is on chromosome 21. This enzyme catalyzes the reaction of serine and homocysteine to cystathionine. Subsequently cystathionine is converted to cysteine. Since this enzyme will be present in excess in a Down Syndrome patient, we would expect a reduction in the substrates serine and homocysteine and an increase in the products cystathionine and cysteine. Referring to Tables 5 and 6, serine in a Down Syndrome patient (Mean=15.1087, Median=13) is reduced from levels in a normal patient (Mean=19.7875, Median=17.25). Cystathionine is greater in a Down Syndrome patient (Mean=0.2087, Median=0.1) than in a normal patient (Mean=0.1288, Median=0.05). Cysteine is greater in a Down Syndrome patient (Mean=212.1957, Median=200) than in a normal patient (Mean=152.9625, Median=144). Homocysteine is greater in a Down Syndrome patient (Mean=0.1565, Median=0.1) than in a normal patient (Mean=0.0625, Median=0.0). 
         [0039]    Prior to the method of the current invention, the aforementioned results would ordinarily be difficult to comprehend. However, pursuant to the invention, the examination of the results from the nutritional molecule FIGLU and from the neurotransmitter normetanephrine enables a treatment to be suggested for these conditions. These results indicate a shortage of mono-carbons. Homocysteine would normally be methylated to methionine and then converted to s-adenosyl-methionine (SAM). SAM is a major source of mono-carbons in the cell. Since such mono-carbons are in short supply, homocysteine accumulates due to the lack of mono-carbons necessary to methylate homocysteine to methionine. Again, this suggests that a supplement of mono-carbons should be prescribed. Thus, a treatment regimen that supplements mono-carbons is prescribed to a patient pursuant to the data provided by the broad metabolic profile analysis described herein. 
         [0040]    Holocarboxylase synthase is located on chromosome 21. This enzyme causes metabolic activation of the vitamin biotin. The data here suggest that some biotin dependent enzymes are accelerated in Down Syndrome. First, beta-lactate, also known as 3-hydroxy-propionate, is a marker for propionyl COA carboxylase, a biotin dependent enzyme. Beta-lactate in a Down Syndrome patient (Mean=6.1522, Median=4.5) is lower than in a normal patient (Mean=11.9625, Median=6). (See Tables 11 and 12). Additionally, leucine is lower in a Down Syndrome patient (Mean=126.5652, Median=93.5) than in a normal patient (Mean=231.5, Median=221.25). (See Tables 5 and 6). Isoleucine is also lower in a Down Syndrome patient (Mean=67.2609, Median=51) than in a normal patient (Mean=103.6875, Median=93.5). Catabolism of both isoleucine and leucine require biotin. 
         [0041]    These results suggest that patients with Down Syndrome have an acceleration of biotin-dependent pathways due to an increase in gene dosage from holocarboxylase synthase. Biotin is generally thought to be non-toxic. If these accelerated pathways were harmful, a patient may benefit from a biotin restriction pursuant to the aforementioned data. 
         [0042]    Another treatment that is suggested by the method of globally analyzing a complete metabolic profile of this patient for chromosomal abnormalities is a tetra-hydra-biopterin supplement. Phenylpyruvate (phenylpyruvic acid) is elevated in a Down Syndrome patient (Mean=0.1435, Median=0.05) over a normal level (Mean=0.0375, Median=0). (See Tables 11 and 12). Phenylalanine is increased in a Down Syndrome patient (Mean=180.2174, Median=150.5) over a normal level (Mean=144.65, Median=138). (See Tables 5 and 6). These two metabolites are elevated in phenylketonuria (PKU), a disease which causes mental retardation. The enzyme affected is phenylalanine hydroxylase. The enzyme requires tetrahydrabiopterin, thought to be deficient in Down Syndrome patients. The aforementioned data suggests that tetrahydrabiopterin deficiency is present in Fetal Down Syndrome and should be treated by prescribing tetrahydrabiopterin supplements. 
         [0043]    Furthermore, analysis of the metabolite data may suggest a vitamin B 6  supplement. Turning again to Tables 11 and 12, oxalic acid (oxalate) in a Down Syndrome patient (Mean=34.8913, Median=23.5) is elevated over normal levels (Mean=12.7, Median=5). This characteristic suggests a functional deficiency of vitamin B6, also known as pyridoxine. Therefore, a vitamin B6 supplement should be prescribed. 
         [0044]    The above findings can be assembled to give an itemized biochemical characterization of fetal Down Syndrome. Some of the biochemical abnormalities are treatable. As shown below, several types of treatment may be prescribed based on the biochemical characterization of Down Syndrome discussed above. 
         [0000]    
       
         
               
               
             
           
               
                   
               
               
                 ABNORMALITY 
                 PRESCRIBED SUPPLEMENT(S) 
               
               
                   
               
             
             
               
                 Mono-carbon Shortage 
                 Folate, B12, Mono-carbon donors (ex. 
               
               
                   
                 Methionine, Betaine, Choline, 
               
               
                   
                 Dimethylglycine) 
               
               
                 Increased Homocysteine 
                 Folate, B12, Mono-carbon donors (ex. 
               
               
                   
                 Methionine, Betaine, Choline, 
               
               
                   
                 Dimethylglycine) 
               
               
                 Increased NE 
                 Folate, B12, Mono-carbon donors (ex. 
               
               
                   
                 Methionine, Betaine, Choline, 
               
               
                   
                 Dimethylglycine) 
               
               
                 Functional B6 deficiency 
                 B6 
               
               
                 Reduced serine 
                 Serine 
               
               
                 Tetra-hydro-biopterin deficient 
                 Tetra-hydro-biopterin 
               
               
                   
               
             
          
         
       
     
         [0045]    With the current method, a simultaneous assessment of all metabolites improves the assessment of individual metabolites and suggests treatment that might have otherwise been overlooked. 
         [0046]    The particular examples set forth herein are instructional and should not be interpreted as limitations on the applications to which those of ordinary skill are able to apply this invention. Modifications and other uses are available to those skilled in the art which are encompassed within the spirit of the invention as defined by the scope of the following claims.