DNA ploidy and expression of P53, cMyc in childhood leukemia's and their families

Chromosomal abnormalities in childhood leukemia have important biological, diagnostic and prognostic significance. The genes that are associated with the development of malignancy were categorized as oncogenes and tumor suppressor genes. P53 belongs to the category of tumor genes and is located on the short arm of chromosome 17 p13. It is a specific transcriptional activator of genes controlling GI checkpoint of the cell cycle and controls the expression of certain genes involved in the control of programmed cell death [Apoptosis]. cMyc gene is Juxtaposed with one of the immunoglobulin genes: heavy chain on 14[q32], kappa on 2[p12] or Iambda on 22[q11]. In pediatric ALL with translocation of ch.,8 [q24] onch., 14[q32] or ch22[q11], c-Myc expression is markedly deregulated by the highly active immunoglobulin locus. This leads to an increase in c-Myc max dimmers and transactivation of multiple cognate target genes, driving uncontrolled cellular proliferation. This study was designed to determine the DNA content [ploidy], expression P53 protein and c-Myc protein in children with acute lymphoblastic leukemia as well as in their first degree relatives [parents and siblings] in order to detect the role of these proteins in the developing of leukemia and those at risk of developing leukemia This study was done on 20 infants and children [16 males and 4 females], their age ranged from 2 to 12 years. They were admitted to the Hematology Unit at Mansoura University Children Hospital where they were diagnosed as acute lymphoblastic leukemia [ALL] and were taken at presentation before induction of treatment. Their first-degree relatives were also included in the study [20 fathers, 20 mothers and 44 siblings [23 brothers and 21 sisters]].Twenty healthy persons with negative family /history of cancers, their aye range from 4 to 30 years, were taken as control group. All the studied subjects were subjected to isolation of lymphocyte which staining and fixation within 24 hours from sampling where DNA analysis by flow cytometer was done. The results proved that 60 % of the cases with acute lymphoblastic leukemia were hyper diploid [DWA index >1.0] and 10 % were hypo diploid [DNA index [1.0]. All of their leukemic children were DNA aneuploid while 10 % of tile-studied parents had DNA aneuploid positive cells and 7% of the healthy siblings had DNA aneuploid positive cells. A high significant level of p53 protein in patients with ALL when compared with healthy controls [p<0.0001], a significant difference in between patients with ALL and their first -degree relative regarding P53 protein. Also a significant difference in between the first-degree relative and healthy controls regarding p53 [p=<0.0001]. The results of this study also showed significant difference in the level of expression of p53 protein among patients with DNA aneuploidy cells compared to those with DNA diploid patients. A significant difference in patients with ALL when compared with healthy controls, also were was significant difference in first degree relatives of leukemia patients versus to healthy controls regarding the level of c-Myc protein [p=<0.0001]. 80 % of parents and 77 % of siblings between the first-degree relative expressing high levels of cMyc protein

Conclusion: genetic alterations play an important role in the development of childhood leukemia particularly mutation of p53 tumor suppressor gene and cMyc gene where the level of expression of their related protein is high. Also change in the DNA content of the lymphoblast's of majority of the patients with acute lymphoblastic leukemia appears to be constant feature. These changes not only present in cases with ALL but also in their first degree relatives suggesting that there is a vertical transmission of these genes in these families. First -degree relatives of leukemia patients particularly those with abnormal DNA and those expressing high level of p53 protein and cMyc protein are at high risk of developing cancers must be subjected for close follow up

Apoptosis in neonatal hypoxic ischemic encephalopathy

The cell death phenomenon is an important feature in the development of the nervous system. Many neurological diseases such as hypoxic ischemic encephalopathy are characterized by the gradual loss of specific sets of neurons and result in disorders of movements and CNS functions. The mechanism of apoptotic neurodegeneration in thalamus and other brain regions after neonatal hypoxic ischemia are completely unknown. However, death receptor-activated pathways, altered mitochondria1 function and changes in expression of mitochondria/-related bcl-2 family proteins are likely important effectors of programmed cell death. Fas /Apo-1[CD95] is an important one of mitochondria/-related bcl-2 family proteins. Fas death receptor [CD95] expression increased in hippocampus bilaterally during 24 hours immediately after neonatal hypoxic ischemic encephalopathy. This study was designed to evaluate the apoptotic process in CNS tissue, which occurred in neonatal hypoxic ischemic encephalopathy by measuring the levels of Fas in serum and CSF. The study was done on 80 neonates. Sixty neonates with hypoxic ischemic encephalopathy and 20 healthy neonates [as control group] were admitted in premature and IC units in Al-Minya university hospital. All neonates were subjected to careful history of HIE, clinical examination with assessment of severity of HIE by using Apgar and Thompson score. Laboratory investigations were done for all neonates and included complete blood picture, C - reactive protein, urine analysis, culture, and CSF by lumbar puncture. Fas/Apo-1[CD95] was measured in serum and CSF by a solid-phase sandwich, two-site, enzyme-linked immunoassay [ELlSA]. The results showed that serum soluble Fas levels in neonates with HIE were significantly elevated [15.81 +/- 7.13 ng/dl] compared with control group [4.09 +/- 1.15 ng/dl] [p< 0.0001]. Significant increase in soluble Fas levels in CSF was detected in HIE neonates [3.93 +/- 3.01 ng/dl] when compared with healthy control neonates [0.33 +/- 0.27 ng/dl] [p>0.0001]. There was also a significant positive impact of the prematurity on Fas in serum and CSF, where a statistically significantly increase in serum and CSF Fas levels was reported in HIE premature neonates [210.9 +/- 5.59 ng/dl][6.62 +/- 2.73 ng/dl respectively for serum and CSF], compared to those of HIE full-term neonates [12.76 +/- 5.76 ng/dl, 2.5and 2.11 ng/dl respectively] [p<0.0001]. There was also a significant correlation between the levels of Fas in serum and CSF with the severity of HIE according to Thompson score

Conclusion: the increase in serum and CSF Fas levels of HIE in neonates explain the apoptotic process in brain tissue damage in HIE. Measuring Fas levels in serum and CSF of HIE neonates can be used as marker of apoptosis and can detect the severity of HIE

Plasma levels of nerve growth factor and transforming growth factor-beta2 in insulin dependant diabetes mellitus in children

This study was designed to evaluate the role of nerve growth factor [NGF] and transforming growth factor-beta2 [TGF-beta2] in the pathogenesis of insulin dependant diabetes mellitus [IDDM]. This study was done on diabetic pediatric patients regularly visiting the pediatric out-patient clinic of diabetes at El Minia university hospital. Nerve growth factor and transforming growth factor-Beta2 levels were measured in the serum of the patients with insulin dependant diabetes mellitus [n = 20] and the resulted values were compared to those of age-matched normal subjects [n = 13]. All IDDM patients were on multiple daily insulin injections and on no other medications. Both nerve growth factor and transforming growth factor-Beta2 were measured by a solid-phase sandwich, two-site, enzyme-Iinked immunoassay [ELISA]. The results proved that serum nerve growth factor levels were significantly elevated in insulin dependant diabetes mellitus patients [1798 +/- 296 pg/ml] compared to those of age-matched control [687 +/- 145pg/ml]; [p < 0.001]. Serum transforming growth factor-Beta2 levels were significantly decreased in IDDM patients [294 +/- 48pg/ml] when compared with the healthy control [577 +/- 74pg/ml]; [p <. 001]. There was no correlation between the levels of nerve growth factor and transforming growth factor-Beta2

Conclusion: We concluded that an increase in nerve growth factor and decrease in transforming growth factor-beta2 levels were present in patients with insulin dependant diabetes mellitus and that both cytokines may play independent roles in the pathogenesis of this disease

Cytogenetic evaluation of epilepsy

The aim of this study was to detect the chromosomal abnormalities in patients suffering from epilepsy through chromosomal study, and to evaluate the correlation between these results with physical and intellectual disabilities. The present study was conducted on 50 epileptic pediatric patients [29 males and 21 females], their age ranged between 6 months and 11 years. Patients were selected from Pediatric Neurology out-patient clinic in EI-Minia University Hospital, from December 2002 to August 2003. All patients were diagnosed as having epilepsy based on the detailed clinical symptomatology of seizures. The patients were subjected to careful history taking, complete general and neurological examination. Electroencephalography [EEG], computerized tomography of brain [C.T.] and chromosomal study by studying G-bands to facilitate the identification of structural abnormalities were done. The studied patients were divided into 2 groups: Group 1 included 20 epileptic patients with normal physical and intellectual developments and group 2 included 30 epileptic patients with intellectual disabilities and/or physical abnormalities. The results proved that 4 cases [8%] showed chromosomal abnormalities among the group 2 of patients. One patient had distal deletion of the long arm of chromosome 1, one patient had ring chromosome 20 and the remaining two patients had distal deletion of the long arm of chromosome 6

Conclusion : We concluded that epileptic patients not in need for chromosomal study, karyotype analysis should be performed in a child or adult complained from seizures with dysmorphic features and/ or intellectuaidisability

Clinical and biochemical studies on the effect of glucose insulin potassium infusion during open heart surgery for mitral valve replacement

The metabolic and homodynamic effects of glucose insulin potassium infusion for patients undergoing mitral valve replacement, has been studied in 30 adult non diabetic patents divided into two groups, fifteen each. The first group of patients were given glucose insulin potassium infusion [GIK] in a dose of 0.5 g kg[-1] hr[-1] of glucose to unit kg[-1] hr[-1] of insulin to 0.25 mmol kg[-1] hr[-1] potassium. The infusion started after induction of anacsthesia till the aortic cross-clamping. The second group of patients were given lacted ringer infusion at the same period of study as a control group. The anaesthetic technique and drugs were standardized in all patients, well as the cardiologia and extracorporeal techniques to climinate other hemodynamic effects. Comparison between both groups showed significant lower incidence of seirous arrhythmia in patients receiving GIK infusion [P< 0.05]. The need for D.C. shocks to defibrillate the heart and the inotropic support after the bypass were also lowered. The hemodynamic monitoring of both groups was achieved by pulmonary artery catheres. Invasive arterial blood pressure and ECG monitoring. The hemodynamic comparison showed a non significant change in heart rate, C.V.P. and S.V.R. ad arterial blood pressure in both groups, while the P.A.P. and P.C.W.P decreased significantly in GIK group [P< 0.05]. The cardiac output and cardiac index creased significantly [P< 0.001] in GIK group in the after bypass and postoperative period. The cardiac index was increased significantly in GIK group especially [P< 0.001]. The scarified value was taken and a piece of papillary, muscle was biopchemically analysed., the glycogen ATP and LDII levels were risen significantly [P< 0.001] in GIK group relative to the control group. Better myocardial performance was found in patients using GIK infusion before bypass. The routine use of infusion can be recommended for all patients undergoing open heart surgery