High expression of the wild type of ALK gene is associated with favorable prognosis in human neuroblastomas

Anaplastic lymphoma kinase [ALK] gene is a transmembrane receptor tyrosine kinase initially discovered as part of the NPM-ALK fusion protein, resulting from a chromosomal rearrangement frequently associated with anaplastic large cell lymphomas. ALK is a novel dependence receptor and normally expressed in the developing and adult nervous system. The mutated ALK gene has been identified as a potential oncogene in human neuroblastomas [NBLs]. However, the frequency of mutation is only 5-8%. The present study was performed to examine the level of ALK mRNA gene expression in primary neuroblastoma patient tumor tissue samples and to assess the relation between ALK gene expression and other previously reported prognostic factors of neuroblastoma. Methods: Quantitative real-time RT-PCR was applied to examine the expression level of ALK mRNA. and its prognostic value in primary neuroblastoma patients was analyzed by the statistical methods. Immunohistochemical staining was used to check the expression level of ALK proteins. In analysis of 79 patients with sporadic primary neuroblastoma, we found that high expression level of ALK mRNA was significantly associated with Shimada's pathological classification [p=0.0000], patient's age [p-0.0000], MYCN amplification status [p=0.0000], tumor stage [p=0.0000][and] TrkA expression level [p=0.0390], all these factors are known to be associated with good prognosis in neuroblastoma. Of interest, immunohistochemical study revealed positive ALK in ALK-ampllfizd tissues and cell lines. Furthermore, mutation results showed that ALK mutation represented about 4.6% of cases and ALK amplification represented about 1.5% of cases. So that mutations not only occur among unfavorable cases with low ALK but also in favorable cases with high ALK expression. Our findings suggested that, high expression of wild type ALK is associated with good prognosis of NBL and that, as the mutation and amplification of ALK are reported to play an important role in NBL, also the expression level of wild type ALK gene might also have some function in cell growth as well as differentiation in neuroblastoma

Impact of HLA-C matching on acute graft-versus-host disease in allogenic hematopoietic stem cell transplantation

Outcome of unrelated donor marrow transplantation is influenced by donor/recipient matching for HLA. Prior studies assessing the effect of mismatch at specific HLA loci have yielded conflicting results. Disparity for HLA-A or HLA-B antigens increases the risk of poor marrow graft outcome, but little is known about the relevance of HLA-C matching. This work aimed to evaluate the effect of HLA-C matching on hematopoietic stem cell transplant, outcome specifically on the acute graft versus host disease [aGVHD]. Seventy patients given hematopoietic stem cell transplant [HSCT] in different transplant centers with their relevant donors were included in the study, HLA class I [HLA-A, -B] and class II [DRB1, DQR1, DPB1] typing was performed using polymerase chain reaction-sequence-specific oligonucleotide probe [PCR-SSOP] and HLA-C typed by PCR-sequence specific priming [PCR-SSP] technique. The risk of aGVHD during the first three months after HSCT was estimated. Fifty two [74.3%] donor/recipient pairs were mismatched for HLA class I alleles, eighteen [34.6%] of them experienced aGVHD. While no history ofaGVHD was reported in eighteen HLA class I full matched pairs [25.7%]. Higher incidence of aGVHD was observed with isolated HLA-A mismatch [28.6%], and for isolated HLA-B and HLA-C locus mismatch, the incidence were 25% and 16.7% respectively. The incidence of aGVHD was elevated if HLA-A or HLA-B mismatch is associated with HLA-C mismatch [40% and 37.5% respectively]. Much higher incidence of aGVHD was associated with combined HLA-A-B and-C mismatch [44.4%]. The estimated odds ratio [OR] of aGVHD for total HLA-C mismatch relative to matched pairs [univariable model] was 5.0 [95% CI 1.3-19.4; P= 0.01]. The degree of HLA-C allele mismatch [one or two alleys mismatch] were significantly related to aGVHD outcome [OR, 3.9, P= 0.03; OR, 10.8; P .009 respectively]. HLA-A or HLA-B allele disparity was also associated with aGVHD. As regard the analysis of total HLA-A and -B mismatched pairs with their corresponding matched locus, they demonstrated significant adverse effect on aGVHD [OR, 2.8; P=0.04; OR 3.0; P=0.03]. It was concluded that HLA-C may function as a powerful transplantation antigen. HLA-C compatibility should be incorporated into algorithms for donor selection to improve the outcome especially in patients who have an increased risk. The presence of HLA-C mismatch with either HLA-A or HIA-B mismatch leads to a synergistic increase in cytotoxic responses and poor graft outcomes [the development of aGVHD]; however, isolated HLA-C mismatch may be acceptable with respect to T-cell mediated alloreactivity

prognostic value of interleukin-6 [IL-6] and C-Reactive protein [CRP] in non-hodgkin's lymphoma [NHL]

The prognostic value of two simple serum markers [IL-6 and CRP] was prospectively studied in 40 newly diagnosed patients with non- Hodgkin's lymphoma treated with standard CHOP or CVP chemotherapy regimen for high and intermediate grade or low grade, respectively, who fulfilled the criteria for inclusion in the study. In addition, 20 healthy volunteers served as a control group were included in this study. The significance of these markers was compared with other known prognostic indicators by the statistical analysis. The study revealed a strong correlation between IL-6 and CRP in NHL patients as regards the response and prognostic factors. In unvaried analysis, the age, stage, tumor mass, pretreatment serum albumin, LDH, IL-6 and CRP levels were found to have an influence on the remission rate; where IL-6 and CRP were decreased to a normal level in patients who underwent a complete response [IL-6 mean +/- SD = 7.6 +/- 2.3, CRP mean +/- SD = 6.1 +/- 2.80] and increased in relapsed patients [IL-6 mean +/- SD = 34 +/- 8.9, CRP mean +/- SD = 36 +/- 13.21]. However, in the regression analysis to the results of 33 cases with high and intermediate grade, the pre-treatment IL-6 and CRP levels were the only predictors for remission

Serum levels of glutamic acid decarboxylase autoantibodies [GADA], soluble FAS/APO-1 [CD95], interleukin-1beta [IL-1Beta] and nitric oxide [NO] in children with insulin-dependent diabetes mellitus

Although the destruction of pancreatic islet Beta cells is the defining characteristic of autoimmune diabetes, the precise cellular and molecular mechanisms effective for beta cell death have yet to be defined as we are still rather ignorant about the events directly responsible for this death. Markers of apoptosis [lL-1beta and NO], antiapoptotic marker [s-Fas], glutamic acid decarboxylase antibodies [GADA] and albuminuria were studied in 60 insulin dependent diabetic children and in 17 age and sex matched healthy children. Patients were classified into two groups according to the duration of diabetes into: group "A" included insulin dependent diabetics with disease duration < 5 years and group " B" insulin dependent diabetics with disease duration of 5 years or more. When glycemic control was taken into account the diabetic patients were further classified into metabolically controlled diabetics and non metabolically controlled ones according to the level of glycated hemoglobin. Our results revealed that the mean serum levels of IL-1 beta, NO, GADA and urinary albumin excretion rate were significantly higher, while serum levels of s-Fas were significantly lower in all diabetics than in controls, in diabetics with disease duration of 5 years or more than in those with disease duration <5 years and in non metabolically controlled diabetics than metabolically controlled ones. Significant positive correlations were detected between GADA serum levels with each at apoptotic markers, lL-1beta and NO and patients' age, between serum levels of IL-1Beta and NO, as well as between urinary albumin excretion and each of glycated hemoglobin, apoptotic markers [lL-1beta, NO] and GADA. Statistically significant negative correlations were detected between anti apoptotic marker, s-Fas and each of apoptotic markers, GADA and urinary albumin excreted in urine. Sixty four percent of diabetics with disease duration <5 years were normoalbuminuric [n= 28/44] and 36.36 % were microalbuminuric [stage 1 nephropathy] [n=16/44]. While 25% of diabetics with disease duration of 5 years or more were normoalbuminuric [n=4/16], 56.25 % were microalbuminuric [stage 1 nephropathy] [n=9/16] and 18.75 % were macroaibuminuric [stage 2 nephropathy][n=3/16]. No significant increase in the mean levels of blood urea and serum creatinine was detected in all studied diabetic groups when compared with controls and with each other

Conclusion: The present study clearly indicates the role of both lL-beta cytokine and NO as apoptotic markers, s-Fas as antiapoptotic marker and GADA in mediating islet B-celi damage in insulin dependent diabetics as well as their prognostic values. ? albuminuria is an early indicator of early diabetic nephropathy. The levels of apoptotic markers, GADA and excreted urinary albumin predict progression to overt nephropathy. Routine monitoring of all diabetic children for micro albuminuria must be done as it is a simple and reliable technique identifying insulin dependent diabetics at high risk for nephropathy. Apoptotic markers, GADA and albumin excretion rate are aggravated with prolongation of the disease duration and deterioration of glycemic control. Good glycemic control and novel strategies aimed at modifying the apoptotic process or inducing immune tolerance to antigens may arrest or reverse the process of beta-cell death, thereby preventing the onset of IDDM or lessening its clinical severity

Serum levels of mutant p53, soluble FAS [S-FAS] in children with acute lymphoblastic leukemia

In a trial to improve the outcome of acute lymphoblastic leukemic patients, the serum concentrations of mutant p53 and soluble Fas [s- Fas] as antiapoptotic markers were assessed in the sera of 45 children with acute lymphoblastic leukemia [ALL] [on admission, after remission and in relapsed cases] as well as in 15 healthy children of matched age and sex as controls [on admission]. The study revealed a significant increase of mutant p53 and s-Fas in newly diagnosed cases than in the controls and ALL cases in remission, in non responders than in responders and in cases with organomegaly than those without. Although mutant p53 and s-Fas had the same specificity and negative predictive values, mutant p53 had significantly higher sensitivity and positive predictive value than s-Fas in ALL cases with metastases as well as in relapsed cases. On admission, there was a significant positive correlation between blast cells% and each of mutant p53, s- Fas levels as well as between the levels of the last two antiapoptotic markers

Plasma endothelin-1 and thrombomodulin: correlation with the extent of pulmonary hypertension in patients whith chronic hypoxic pulmonary disease

This study included 47 pulmonary hypertensive patients secondary to chronic hypoxia due to chronic obstructive pulmonary diseases [COPD] and interstitial pulmonary fibrosis [IPF]. They were sampled in the morning after an overnight fast, before medication and after termination of treatment. Endothelin-1 level was 1.45 +/- 0.32 pg/ml which was increased compared with the normal controls [0.5 +/- 0.02]; the difference was statistically significant. Endothelin-1 was significantly correlated with pulmonary pressure and its degree of severity, especially among chronic obstructive pulmonary disease patients. There was a significant reduction in the level of endothelin-1 after long-term oxygen therapy; whereas, it did not correlate with the level of PaO2 either before or after oxygen therapy. Plasma thrombomodulin level was measured to study its pathophysiological significance in pulmonary hypertension secondary to hypoxia. Patients with pulmonary hypertension had higher concentrations of thrombomodulin [15.25 +/- 1.5] than the age matched normal controls [4.8 +/- 0.5 ng/ml]. There was a significant reduction in thrombomodulin after oxygen therapy. Plasma concentrations of thrombomodulin were significantly correlated with time to peak velocity [TPV] as a strong echo parameter of pulmonary hypertension and its severity in mild PH thrombomodulin level was 10.46 +/- 0.56 ng/ml versus 15.82 +/- 3.08 ng/ml among patients with severe pulmonary hypertension. Moreover, thrombomodulin correlated with PaO2 after oxygen therapy especially with patients of IPF

Antithrombin III, heparin cofactor II and histidine-rich glycoprotein in patients with pulmonary embolism

In seventy-two patients hospitalized for clinical suspicion of pulmonary embolism [PE], level of antithrombin III [AT III], heparin cofactor II [HC II] and histidine-rich glycoprotein [HRGP] were determined. Forty-four patients were positive for PE and twenty-nine received standard heparin. HC II and HRGP did not show any statistically significant variation between patients who were positive or negative for PE. Moreover, presence or absence of heparin therapy in these patients did not modify the level of these two proteins. In contrast, AT III showed a statistically significant reduction in PE than non-PE. Since AT III is one of the most powerful inhibitors against serine protease enzymes of coagulation cascade, it is possible that low levels in patients with PE could be considered as the expression of consumption to prevent further thrombus formation

Some clinico-laboratory aspects of the problems of neonatal jaundice

This study included one hundred newborns with neonatal jaundices as well as twenty age and sex matched newborns as a control group. The detected causes of jaundice in this study were isoimmunization [Rh and ABO] [33%] and all cases were investigated for the presence of irregular antibodies, but with negative results. The second major cause of jaundice in the study was septicemia[15%], some other causes were found to play a minor role. Thyroid function test [T3, T4 and TSH] was evaluated for detection of hypothyroidism as a cause of neonatal hyperbilirubinemia and all cases were normal for age. Infant risk factors that were recorded among newborns with neonatal hyperbilirubinemia were also studied. The rate of these risk factors were as followed; males sex [72%], prematurity [20%] and intrauterine growth retardation [IUGR][12%]. Most of these cases recovered by treatment of the cause and phototherapy [75% versus 25% of phototherapy and exchange transfusion]. However, 78.5% of cases with Rh-incompatibility and 31.5% of cases with ABO-incompatibility needed exchange transfusion combined with phototherapy. Concerning the outcome of the studied cases, 82% were cured, 13% died and 5% developed complications. It was concluded that proper antenatal care for the pregnant mothers, early diagnosis and management of neonatal jaundice and keeping in mind the possibilities of congenital hypothyroidism are recommended

Renal impairment in jaundiced full term neonates

This study included eighty-five full term newborns with unconjugated hyperbilirubinemia due to ABO and RH incompatibility as well as twenty healthy age and sex matched full term newborns as a control group. None of the cases showed any other stress factor which may cause renal failure e.g. septicemia, asphyxia or diabetic mother. Abdominal ultrasonography was done to exclude cases renal abnormalities. All cases were subjected to full clinical examination. Laboratory investigations of blood samples were taken at the 15th day then at the 30th day of postnatal life for blood culture, Coomb's test, blood urea and serum sodium, potassium, creatinine and uric acid. Blood groups were determined for babies and their mothers. Urine samples were taken at the 5th, 15th and 30th day of postnatal life for urine analysis, total protein, microalbumin, B2-microglobulin, urinary creatinine, sodium and potassium

Urinary B-2 microglobulin and retinol binding protein and microalbuminuria in perinatal asphyxia

This study was carried out on twenty-eight full term neonates with perinatal asphyxia with average one, five and ten minutes Apgar scores of 1.6, 4.3 and 3.7, respectively. Ten full term healthy neonates were included as a control group with one, five and ten minutes Apgar scores of 7.3, 9.4 and 9.6, respectively. Both groups were tested on the second and fifth days of postnatal life as regards complete blood picture, complete urine analysis, culture and sensitivity for urine and blood, investigations for renal glomerular function included urinary concentration of albumin, blood urea and serum creatinine, investigations for renal tubular function included urinary beta-2-microglobulin [B2-m] and retinol binding protein [RBP], fractional excretion of sodium [FENa] and urine output per unit body weight per hour [ml/kg/hr]. Abdominal sonography was done for all cases to exclude any surgical problems. Renal failure index [RFI] was calculated at second and fifth days of postnatal life. It was concluded that measurement of urinary B2-m and RBP are convenient, sensitive, specific and early diagnostic tools for the detection of tubular damage in asphyxiated neonates. Blood urea and serum creatinine are good indicators of glomerular impairment only after the first two days of life. The tubular function is more sensitive than glomerular function in neonates. A low Apgar score, arterial PO2 and pH are good indicators of hypoxia. Oliguria is already considered as an ambiguous indicator of ARF and could indicate poor neurologic outcome in asphyxiated neonates

Acute renal failure in jaundiced-isoimmunized full term neonate

This study included 85 full-term newborns with unconjugated hyperbilirubinemia, due to ABO and RH incompatibility and renal impairment as well as 20 healthy, age and sex matched full-term newborns as a control group. None of the cases showed any stress factors which may cause renal failure. All cases were subjected to full clinical examination, abdominal ultrasonography to exclude cases with congenital renal abnormalities, laboratory investigations of blood were done at the 5th day then repeated at 30th day of postnatal life for follow up. According to the investigations, the patients were subdivided into 2 groups: 25 patients with acute renal failure [ARF] and 60 patients without acute renal failure [non ARF]. They represent 4.2% and 10.1%, respectively. of the total admissions of the jaundiced, isoimmunized full-term newborns in the NICU in one year [596 neonates]. Follow up studies for one month revealed that renal functions were normalized in cases with non-ARF except the beta 2 microglobulin, while cases with ARF still have significantly higher differences in all the studied parameters when compared to the control group after one month except for urine pH and specific gravity. In conclusion, renal impairment should be expected in cases having total serum bilirubin >25 mg/dL. They should be closely monitored regarding their renal functions for at least one month of postnatal life. Those who develop ARF should be followed up for a longer period