Identification of five common Alpha-thalassaemia genotypes among a group of Egyptian neonates by single tube PCR

The alpha thalassaemias are common genetic disorders that arise from reduced synthesis of the alpha globin chains. At present, large scale carrier screening and clinically valuable antenatal detection programs have not been established for the congenital disorder alpha thalassaemia. This study was conducted on 410 umbilical cord blood samples to detect the relative frequency and differentiate between the five common alpha thalassaemia deletional forms, regardless of the break points, among a group of Egyptian neonates using a single tube multiplex-PCR assay. The method proved to be simple, easy and fast as the number of alpha genes present in the subjects were directly determined by the number and intensity of the alpha l and alpha 2 bands normalized with that of beta actin [serving as an internal control]. A complete blood picture was done for all samples. Among the DNA samples tested, ten cases were discarded due to failure of amplification. Our results showed that 38 samples [9.5%] were identified as carriers of alpha thalassaemia or suffering from Hb H disease. The percentage of the different genotypes detected was: 4.5% alpha thalassaemia 2 heterozygous [-alpha/ alpha alpha], 3% alpha thalassaemia 2 homozygous [-alpha /-alpha], 1.75% alpha thalassaemia 1 heterozygous [--/alpha alpha] and 0.25% Hb H disease [--/-alpha]. No cases of Hb B art's hydrops fetalis were detected. On comparing the hematological profile of the normal subjects with that of those with gene deletions it was found that alpha thalassaemia carriers had significantly lower Hb levels, Hct%, MCH and MCHC while the MCV did not differ markedly. No statistically significant difference was found between the various genotypes detected regarding their blood indices. From this we can not rely upon blood indices only for detection of alpha thalassaemia and we highlight the importance of molecular diagnosis using this simple single tube PCR assay in diagnosis of alpha thalassaemia carriers

Transcutaneous bilirubinometery: its role in the assessment of neonatal jaundice with comparable precision with serum bilirubin

Transcutaneous bilirubinometry [TCB] provides a noninvasive, cost effective screening method for significant neonatal jaundice. The aim of this study is to comment on the usefulness of the jaundice meter as a non invasive screening device for hyperbilirubinemia in neonatal intensive care units. The study included 40 neonates suspected on clinical grounds to have neonatal jaundice, admitted to the unit for either prematurity or other clinical conditions. Our results showed a correlation coefficient between total serum bilirubin and the mean of transcutaneous bilirubinometry of the forehead and sternum at 24 and 48 hours of appearance of jaundice to be 0.629 and 0.823 respectively with a significant P value of [0.07. Also, the correlation coefficient between assayed total serum bilirubin level and expectant serum bilirubin level at 24 and 48 hrs. of appearance of jaundice was 0.599 and 0.863 respectively with a significant P value of <0.01

Conclusion: transcutaneous bilirubinometry is a well-established screening method for neonatal jaundice, yet it has limitations of use due to the sensitivity of the method for defecting variation in melanin content of the skin. Also TCB monitoring should not be used to follow babies who have undergone exchange transfusions nor solely relied up to follow infants undergoing phototherapy

Immunophenotyping, MIB-1, p53 and mast cell tryptase in large cell non-Hodgkin's lymphoma

The aim of the present study was to explore by immunohistochemical methods, on lymph node and infiltrated bone marrow bioposies. the potential value of MIB-l proliferation rate. p53 overexpression and Mast cell tryptase [MCT] as possible prognostic fators for large cell lymphomas on 69 adult patients for whom paraffin blocks, clinical data and survival information were available. In the reviewed series, large cell lymphoma included: 84.1% large B-cell lymphoma. 11.6% peripheral T-cell lymphoma and 4.3% anaplastic large cell lymphoma. MIB-1 labeling was quantitated by image analysis and cases were classified as either of low or high proliferation rate taking MIB-l count of 50% as a cutoff value. For p53, nuclear immunoreactivity equal to or more than 20% were considered overexpression or positive. As for MCT, presence of> 5 mast cells/hpf was recorded as high count while counts <5 mast cells/hpf were considered low counts. In the L.N. biopsies examined the mean MIB-l labeling rate was 48.5%. p53 positive tumors contributed 33.3% of cases while high MCT counts were detected in 43.5% of cases. High MIB-l rate, p53 positivity and MCT counts showed a statistically significant relation to high lPl and were associated ".ith poor response to therapy and unfavorable 2-year overall survival and hence were considered risk factors. Additionally, high MCT count was found to show a strong relation with T-cell phenotype and extranodal forms. Bone marrow biopsies were examined to detect infiltrated cases. B.M. infiltration was detected in 15 cases whose levels of MIB-l and p53were in accordance to the corresponding L.N. values.MCT counts in B.M. biopsies were within the high count group but were lower than corresponding L.N. values and were strongly related to T-cell phenotype. It is concluded that MIB- 1. p53 and MCT are valuable prognostic factors which could serve as a guideline for treatment by identifying unfavorable cases for more intensive therapy