p53 mutation in HCV-genotype-4 associated hepatocellular carcinoma in Egyptian patients

HCV-associated hepatocellular carcinoma (HCC) is a common neoplasm in Egypt where genotype-4 is prevalent. In the present study the incidence and pattern of p53 mutations was assessed in relation to HCV-genotype- 4 in Egyptian HCC patients. We investigated 25 HCV positive HCCs for p53 mutations/overexpression in relation to HCV-NS3 by immunohistochemistry, SSCP and sequencing. Genotyping was done using LiPA-II and TRUGENE 5' NC' sequencing kit. Results were correlated to standard clinicopathologic prognostic factors for HCC. Thirteen cases showed p53 overexpression, and 10 showed p53 mutation (13 mutations) by sequencing (72% concordance). The highest mutation rate was in exons 6 and 7 (30%) followed by exons 5 and 8 (20%). Mutations included 3 transitions, 5 transversions, 3 deletions, and 2 insertions. All exon 7 mutations were at codon 249 specific for AFB1 (AGG-->AGT, Arg-->Ser) and codon 248 specific for vinyl chloride contamination (CGG-->TGG, Arg-->Trp). Other mutations reported are novel. Immunostaining for HCV NS3 was detected in 19 cases independent of p53 mutation. p53 aberrations were significantly associated with poor prognostic factors for HCC. However, no specific pattern for p53 mutations was observed in HCV genotype 4-associated HCC and no significant relation between p53 mutations, HCV-NS3 expressions or any HCV sub-genotype-4 sequence.     

Protection by turmeric and myrrh against liver oxidative damage and genotoxicity induced by lead acetate in mice

The effects of lead acetate in the diet (0.5% w/w) on reduced GSH, activity of phase II metabolizing enzyme glutathione S-transferase (GST), lipid peroxidation in liver homogenate and bone marrow chromosomes of mice simultaneously supplemented with powdered turmeric and myrrh for 8 weeks were investigated. Five groups of Swiss male albino mice, each of 30 mice, the first group received a basal diet and served as negative control, the second group received basal diet supplemented with lead acetate only and served as positive control. The other three groups received basal diet supplemented with lead acetate and 1% or 5% turmeric powder and 1% myrrh powder, respectively. Results revealed a significant decrease in the amount of GSH in all treated groups compared with negative control. Also, the activity of GSH S-transferase was significantly decreased in positive control compared with other groups. However, co-administration of the protective plants resulted in a significant increase in the activity of GST compared with both positive and negative control groups. Furthermore, lipid peroxidation was significantly increased in positive control alone, while co-treatment with the protective plants resulted in reduction in the level of lipid peroxidation by 31% and 49% in mice receiving 1% and 5% turmeric powder respectively and 45% in 1% myrrh treated when compared with their respective positive control group. Lead genotoxicity was confirmed through significant reduction in the number of dividing cells, increased total number of aberrant cells and increased frequency of chromosomal aberrations. Simultaneous treatment with these plants significantly reduced the genotoxicity induced by lead administration and the powerful protection was observed with 5% powdered turmeric. It may be concluded that turmeric and myrrh are useful herbal remedies, especially for controlling oxidative damages and genotoxicity induced by lead acetate intoxication.     

IL-13 gene expression in patients with atopic dermatitis: relation to IgE level and to disease severity

Atopic dermatitis (AD) is a chronic inflammatory skin disease frequently associated with an increased serum IgE level. T helper cells are thought to play an important role in the pathogenesis of AD. It is commonly believed that allergens activate Th2 cells, and it is likely that the cytokines produced by Th2 cells are crucial factors in the induction and maintenance of the disease. IL-13 is one of the cytokines that are produced by Th2 lymphocytes and, like IL-4, it can induce the production of IgE. In order to evaluate its role in the pathogenesis of AD, IL-13 mRNA expression was studied in peripheral blood of patients with different degrees of AD and compared with healthy subjects. Also, we correlated its level of expression with the level of serum IgE and with the severity of the disease. EDTA blood was obtained from 25 patients (divided into three groups ranged from mild to severe AD) and 12 normal subjects as a control group. We examined the blood sample for IL-13 mRNA expression using RNA extraction technique, RT-PCR, PCR amplification using primers specific for IL-13 and beta- actin (as internal control) this is followed by visualization of the expressed bands using gel electrophoresis and DNA marker. Serum IgE level was detected using an ELISA kit. Our results revealed that, IL-13 mRNA is significantly expressed in patients with AD as compared to normal control (P<0.001). IL-13 mRNA shows higher level of expression in severe AD group in comparison with both moderate and mild groups (P = 0.05). Serum levels of IgE showed highly significant increase in patients with AD as compared with the control group (p=0.019), its level is significantly higher in severe AD group versus moderate and mild AD groups (P=0.009 and 0.022, respectively). There is a highly significant positive correlation between serum levels of IgE and the levels of IL-13 mRNA expression in all AD groups (P=0.001). In conclusion, the high level of IL-18 mRNA expression in AD, and its correlation with serum level of IgE and with severity of disease indicates that IL-13 is involved in the pathogenesis of the disease and is an important in vivo IgE inducer.     

Impact of C-reactive protein test results on evidence-based decision-making in cases of bacterial infection

ABSTRACT: BACKGROUND: C-reactive protein (CRP) is widely used to detect bacterial infection in children. We investigated the impact of CRP test results on decision-making and summarized the evidence base (EB) of CRP testing. METHODS: We collected information from the hospital records of 91 neonates with suspected sepsis and of 152 febrile children with suspected infection on the number of ordered CRP tests, the number of EB-CRP tests, and the impact of the test results on decision-making. CRP diagnostic accuracy studies focusing on pediatric infections were reviewed critically. The main outcomes were the proportion of CRP tests that were EB and the proportion of tests that affected decision-making. A secondary outcome was the overall one-year expenditure on CRP testing. RESULTS: The current EB for CRP testing in pediatric infections is weak and suggests that CRP is of low diagnostic value. Approximately 54.8% of tests performed for suspected neonatal sepsis and 28% of tests performed for other infections were EB; however, the results of only 12.9% of neonatal sepsis tests and of 29.9% of tests on children with other infections informed decision-making. The one-year overall cost for CRP testing and related health care was $26,715.9. CONCLUSIONS: The routine ordering of CRP for children with infections is based on weak evidence. The impact of the CRP test results on decision-making is rather small, and CRP ordering may contribute to unnecessary health care expenditures. Better quality research is needed to definitively determine the diagnostic accuracy of CRP levels in children with infections.     

Autozygome maps dispensable DNA and reveals potential selective bias against nullizygosity

PurposeCopy number variants are an important source of human genome diversity. The widespread distribution of hemizygous copy number variants in the DNA of healthy humans suggests that haploinsufficiency is largely tolerated. However, little is known about the extent to which corresponding nullizygosity (two-copy deletion) is similarly tolerated.MethodsWe analyzed a cohort of first cousin unions to enrich for shared parental hemizygous events and tested their Mendelian inheritance in offspring.ResultsAnalysis of autozygous DNA blocks (autozygome) in the offspring not only proved an efficient method of mapping “dispensable” DNA but also revealed potential selective bias against the occurrence of nullizygous changes. This bias was not restricted to genic copy number variants and was not accounted for by a high rate of miscarriages.ConclusionsThe autozygome is an efficient way to map dispensable segments of DNA and may reveal selective bias against nullizygosity in healthy individuals.Genet Med 2012:14(5):515–519     

Multiple intra-familial transmission patterns of hepatitis B virus genotype D in north-eastern Egypt

The transmission rate of intra‐familial hepatitis B virus (HBV) and mode of transmission were investigated in north eastern Egypt. HBV infection was investigated serologically and confirmed by molecular evolutionary analysis in family members (N = 230) of 55 chronic hepatitis B carriers (index cases). Hepatitis B surface antigen (HBsAg) and hepatitis B core antibody (anti‐HBc) prevalence was 12.2% and 23% among family members, respectively. HBsAg carriers were prevalent in the age groups; <10 (16.2%) and 21–30 years (23.3%). The prevalence of HBsAg was significantly higher in the family members of females (19.2%) than males (8.6%) index cases (P = 0.031). HBsAg and anti‐HBc seropositive rates were higher significantly in the offspring of females (23%, 29.8%) than those of the males index cases (4.3%, 9.8%) (P = 0.001, 0.003), as well as higher in the offspring of an infected mother (26.5, 31.8%) than those of an infected father (4.7%, 10.5%) (P = 0.0006, 0.009). No significant difference was found in HBsAg seropositive rates between vaccinated (10.6%) and unvaccinated family members (14.8%). Phylogenetic analysis of the preS2 and S regions of HBV genome showed that the HBV isolates were of subgenotype D1 in nine index cases and 14 family members. HBV familial transmission was confirmed in five of six families with three transmission patterns; maternal, paternal, and sexual. It is concluded that multiple intra‐familial transmission routes of HBV genotype D were determined; including maternal, paternal and horizontal. Universal HBV vaccination should be modified by including the first dose at birth with (HBIG) administration to the newborn of mothers infected with HBV. J. Med. Virol. 84:587–595, 2012. © 2011 Wiley Periodicals, Inc.     

IL-6 promoter polymorphism (?174G/C) and systemic lupus erythematosus

We investigated the possible association between susceptibility to systemic lupus erythematosus (SLE) and single-nucleotide polymorphisms located in the promoter region of the interleukin-6 gene (?174 G/C) in a sample of the Egyptian population and the contribution of this polymorphism in the clinical or immunological manifestation of the disease. Forty-two Egyptian patients with SLE and 40 unrelated healthy control volunteers were genotyped by polymerase chain reaction followed by visualization on 4% agarose gel electrophoresis on ultraviolet transilluminator to detect the genotype distribution and allelic frequencies of the polymorphisms. The homozygous GG genotypes was significantly increased in SLE patients compared to control group (p value?=?0.04). On the other hand, the heterozygous G/C genotype was significantly elevated in the controls compared to SLE patients (p value?=?0.01). The odds ratio value for G/G was 2.6 with a 95% CI from 1.1 to 6.7. As regard the association of clinical manifestations to the genotype frequency, we found a statistical significant increase in the frequency of GG genotype with chest disease, nephritis, and arthritis. From this study, we suggest that G carrier is more susceptible to develop SLE and that SNP may have a role in the pathogenesis of the disease and may be associated with some of its clinical manifestations.     

Widespread cortical thinning in children with frontal lobe epilepsy

Purpose: Spread of seizure activity outside the frontal lobe due to cortico‐cortical connections can result in alteration in the cortex beyond the frontal lobe in children with intractable frontal lobe epilepsy (FLE). The aim of this study was to identify regions of reduced cortical thickness in children with intractable FLE. Methods: High‐resolution volumetric T₁‐weighted imaging was performed on 17 children with FLE, who were being evaluated for epilepsy surgery, and 26 age‐matched healthy controls. The cortical thickness of 12 patients with left FLE and 5 patients with right FLE was compared to controls. The clusters of cortical thinning were regressed against age of seizure onset, duration of epilepsy, seizure frequency, and number of medications. Key Findings: In children with left FLE, cortical thinning was present in the left superior frontal, paracentral, precuneus, cingulate, inferior parietal, supramarginal, postcentral, and superior temporal gyri, as well as in the right superior and middle frontal, medial orbitofrontal, supramarginal, postcentral, banks of superior temporal sulcus, and parahippocampal gyri. In children with right FLE, cortical thinning was present in the right precentral, postcentral, transverse temporal, parahippocampal, lingual, and lateral occipital gyri, as well as in the left superior frontal, inferior parietal, postcentral, superior temporal, posterior cingulate, and lingual gyri. In children with left FLE, following exclusion of one outlier, there was no significant association between age at seizure onset, duration of epilepsy, seizure frequency and number of medications with clusters of cortical thinning. In children with right FLE, age at seizure onset, duration of epilepsy, frequency of seizures, and number of medications were not associated with clusters of cortical thinning within the right and left hemispheres. Significance: Cortical changes were present in the frontal and extrafrontal cortex in children with intractable FLE. These changes may be related to spread of seizure activity, large epileptogenic zones involving both frontal and extrafrontal lobes, and development of secondary epileptogenic zones that over time lead to cortical abnormality. Further studies correlating cortical changes with neurocognitive measures are needed to determine if the cortical changes relate to cognitive function.