1. Methodology Results Amino Acid Levels in Newborns A Pilot Study on an Expanded Newborn Screening Program in Palestine Samir Khatib, Amer Ayyad Medical School / Al-Quds University Introduction Newborn screening is a universal health program which is useful in the early detection of serious metabolic and genetic diseases in newborns before symptoms begin to appear. Until this day newborns in Palestine are being screened for phenylketonuria and congenital hypothyroidism only. However, a number of other metabolic diseases have been recognized amongst the population, including amino acid and urea cycle disorders. Therefore, the study was conducted in an effort investigate the possibility of finding such cases in newborns throughout the West Bank of Palestine. The study was conducted in collaboration with the Palestinian Ministry of Health through the collection of samples, and the Human Genetics Department at Liege University in Belgium by analysing the samples. Study objectives *To assess the need for expanding the existing newborn screening program in Palestine. *To investigate the relationship between some demographic variables such as gender, kinship, district, weight and amino acids levels. *To determine the reference ranges for a number of amino acids and urea cycle intermediates in Palestinian newborns. A cross-sectional observational study design was done. The study covered all 12 districts of the West Bank in Palestine during the period between January 2012 and January 2013. A pilot study was done before the actual study took place. Convenience sampling was used to recognize the study participants of newborns. Blood samples were collected on special blood collection cards. Information regarding weight, date of birth, sex, district, and kinship of parents was taken. Number of samples was 4240 and informed consent was taken from each parent participant. Analysis of amino acids and urea cycle intermediates was done using tandem mass spectrometry (MS/MS). Introduction and Study objectives Of the 4240 newborns, almost half (50.4%) of the study sample were males. Most of the parents of the newborns (74.3%) were unrelated. The mean weight of the participating newborns was 3.23 +0.45 kg. The reference range for each amino acid was calculated based on the non- parametric percentile method (CLSI C28-A3). Statistical analysis showed significant relationship between mean weight and some amino acids levels. From odds ratios test, neither gender nor kinship had any effect on the amino acid levels. The results also showed that there is a significant difference between some amino acids levels and districts. Amino Acids Amino Acids Lower limit (µmol/L) Upper limit (µmol/L) Upper limit (µmol/L)Mean Std. Deviation Std. Deviation Phenyalanine 34.979.7 53.3111.24 Tyrosine 60.2201.3 114.9935.54 Methionine 11.727.9 18.524.19 Leucine-Isoleucine 127.0336.0 208.9552.72 Valine 82.2206.3 131.4630.96 Citrulline 9.326.2 15.784.30 Argininosuccinate 0.20.8 0.430.16 Arginine 4.828.512.956.19 Asparagine-Ornithine 57.5244.9121.5246.86 Reference ranges of amino acids based on Non parametric percentile method(CLSI C28-A3) Amino Acid Amino AcidNormalAbnormalTotal CountRow N %CountRow N %CountRow N % Phenylalanine 410596.8% 1343.2% 4239100% Tyrosine 408196.3% 1583.7% 4239100% Methionine 407696.2% 1633.8% 4239100% Leucine-Isoleucine 406896.0% 1714.0% 4239100% Valine 407696.2% 1633.8% 4239100% Citrulline 406996.0% 1704.0% 4239100% Argininosuccinate 408996.5% 1503.5% 4239100% Arginine 406996.0% 1704.0% 4239100% Asparagine-Ornithine 407696.2% 1633.8% 4239100% Conclusions *Based on our established reference ranges for each analyte the results showed that 22.5% of the tested samples (955) had at least one amino acid level in the upper 2.5% of the population. * There is a significant relationship between mean weight and gender. *There is a significant relationship between mean weight and some amino acids levels. *Neither gender nor kinship had any effect on the amino acid level, but there is a significant difference between some amino acids mean and gender. *There is a significant difference between some amino acids levels and districts. * The study results support further assessment of the need to expand the existing newborn screening program in Palestine. Contact information Status of Abnormal Amino Acid Levels in Newborns Amino AcidGender NMeanStd. Deviation Std. Error Mean P-Value by independent sample T-Test Phenylalanine Female 2104 53.7314.430.31.714 Male 2135 53.8812.560.27 Tyrosine Female 2104 119.1141.440.90.002 Male 2135 115.2339.390.85 Methionine Female 2104 18.994.820.11.034 Male 2135 18.665.100.11 Leucine-Isoleucine Female 2104 211.1458.291.27.065 Male 2135 214.6264.161.39 Valine Female 2104 134.3134.160.74.180 Male 2135 132.8437.280.81 Citrulline Female 2104 16.395.670.12.090 Male 2135 16.095.880.13 Argininosuccinate Female 2104 0.440.180.00.302 Male 2135 0.450.190.00 Arginine Female 2104 13.037.040.15.000 Male 2135 13.837.650.17 Asparagine-Ornithine Female 2104 124.4362.771.37.289 Male 2135 126.3655.821.21 Relationship between Gender and Mean Amino Acid Levels Amino Acid TestMean Weight P-Value by Independent sample T-Test Phenylalanine Normal 3.23 0.718 Abnormal 3.22 Tyrosine Normal 3.23 0.011 Abnormal 3.13 Methionine Normal 3.23 0.214 Abnormal 3.18 Leucine-Isoleucine Normal 3.24 0.001 Abnormal 3.11 Valine Normal 3.23 0.193 Abnormal 3.18 Citrulline Normal 3.23 0.52 Abnormal 3.21 Argininosuccinate Normal 3.23 0.852 Abnormal 3.22 Arginine Normal 3.23 0.367 Abnormal 3.26 Asparagine-Ornithine Normal 3.23 0.06 Abnormal 3.16 Relationship between Mean Weight and Amino Acid Levels Relationship between District and Phenylalanine Level Relationship between District and Citrulline Level Relationship between District and Arginine Level Relationship between District and Asparagine-Ornithine Level Samir Khatib, Genetics and Metabolic Diseases Center, Alquds University Medical School, Abu Dis, Jerusalem, Palestine. skhatib@med.alquds.edu skhatib@med.alquds.edu 00972-54-6352826. NOTE: The authors disclose no conflict of interest.