Enzyme kinetics of 5 alpha-reductase were compared in cultured genital skin fibroblasts taken fro... more Enzyme kinetics of 5 alpha-reductase were compared in cultured genital skin fibroblasts taken from 6 control subjects and from an affected subject with 5 alpha-reductase deficiency in whom the diagnosis was established on hormonal grounds. The Km value for testosterone in the mutant enzyme was extremely high (1,427 vs. 185-417 nmol/l in controls). in the mutant enzyme was extremely high (1,427 vs 185-417 nmol/l in controls). A A mutant but stable enzyme with reduced affinity to steroid substrate is reported.
Two transgenic mice were produced by microinjection of the entire hepatitis B virus (HBV) genome ... more Two transgenic mice were produced by microinjection of the entire hepatitis B virus (HBV) genome as a 3.2-kilobase EcoRI DNA fragment into one-cell embryos. Each animal contained a single, unique locus of HBV sequence. One founder animal, G7, contained a partially deleted HBV genome lacking both putative HBV surface antigen (HBsAg) promoters. The other animal, G26, contained greater-than-genome-length HBV sequences organized as a partial head-to-tail dimer. Both transgenic animals transmitted the HBV sequences in a Mendelian fashion, and all subsequent transgenic animals had detectable HBsAg in the serum. Expression of HBV sequences in tissues from G7- and G26-derived mice showed preferential expression of the 2.1-kilobase HBsAg RNA transcript in liver and kidney tissues by Northern (RNA) blot analysis. These data are consistent with the notion that HBV DNA contains cis-acting regulatory sequences which are responsible for the predominant expression of HBsAg transcripts in the liver...
Molecular genetic analysis of individuals from 6 Egyptian and 33 German families with fragile X s... more Molecular genetic analysis of individuals from 6 Egyptian and 33 German families with fragile X syndrome and 240 further patients with mental retardation was performed applying a completely non-radioactive system. The aim of our study was the development of a non-radioactive detection method and its implementation in molecular diagnosis of the fragile X syndrome. Furthermore, we wanted to assess differences in the mutation sizes between Egyptian and German patients and between Egyptian and German carriers of a premutation. Using non-radioactive polymerase chain reaction (PCR), agarose gel electrophoresis and blotting of the PCR products, followed by hybridisation with a digoxigenin-labelled oligonucleotide probe (CGG)5 and chemiluminescent detection, we identified the fragile X full mutation (amplification of a CGG repeat in the FMR-1 gene ranging from several hundred to several thousand repeat units) in all patients. We observed no differences in the length of the CGG repeat between the Egyptian and German patients and carriers, respectively. However, in one prenatal diagnosis, we detected only one normal sized allele in a female fetus using the PCR-agarose assay, whereas Southern blot analysis with the digoxigenin labelled probe StB 12.3 revealed presence of a full mutation. Our newly established nonradioactive genomic blotting method is based on the conventional radioactive Southern blot analysis. Labelling of the probe StB 12.3 with digoxigenin via PCR allowed the detection of normal, premutated and fully mutated alleles. For exact sizing of small premutated or large normal alleles, we separated digoxigenin labelled PCR products through denaturing polyacrylamide gelelectrophoresis (PAGE) and transfered them to a nylon membrane using a gel dryer. The blotted PCR-fragments can easily be detected with alkaline phosphate-labelled anti-digoxigenin antibody. The number of trinucleotide repeat units can be determined by scoring the detected bands against a digoxigenated M13 sequencing ladder. Our newly developed digoxigenin/chemiluminescence approach using PCR and Southern blot analysis provides reliable results for routine detection of full fragile X mutations and premutations.
Egyptian Journal of Medical Human Genetics, May 1, 2002
... Assessment of the Pattern of Cerebral Blood Flow in Patients with Fragile X Syndrome Using Si... more ... Assessment of the Pattern of Cerebral Blood Flow in Patients with Fragile X Syndrome Using Single Photon Emission Computed Tomography Hanan H. Afifi,1 Alice K. Abd EI-Aleem,2 Laila F. Malta,3 Adel M. Ashour,1 Mostafa K. EI-Awady,2 Nagwa A. Meguid1. ...
Eastern Mediterranean health journal = La revue de santé de la Méditerranée orientale = al-Majallah al-ṣiḥḥīyah li-sharq al-mutawassiṭ
Analysis of expressed mRNAs with differential display-polymerase chain reaction (DD-PCR) is a pow... more Analysis of expressed mRNAs with differential display-polymerase chain reaction (DD-PCR) is a powerful tool for the characterization of genes involved in malignant pathways and might identify markers for different phases of chronic myelogenous leukaemia (CML). We examined the presence of BCR-ABL transcripts in 25 CML patients in either the chronic phase or blast crisis. We then analysed the expression of leukocytic RNA transcripts in CML phases. DD-PCR technique was used to examine CML cases with BCR-ABL in comparison with CML cases lacking detectable BCR-ABL transcripts. Our results support the use of differential display not only for characterization of the CML differentially expressed genes but also to locate patterns that can be implemented as valuable fingerprints for each phase of CML.
Nature and Science, 6(2), 2008, ISSN: 1545-0740, naturesciencej@gmail.com ... CTT Multiplex Syste... more Nature and Science, 6(2), 2008, ISSN: 1545-0740, naturesciencej@gmail.com ... CTT Multiplex System is a quick and inexpensive method to exclude innocent suspects during criminal inquiries ... Ahmed M. Refaat1, Wael T. El-garf2, Kholoud S. Ramadan3, Mohamed M. ...
Our aim was to determine the level of tetanus immunity in the general Egyptian population. The su... more Our aim was to determine the level of tetanus immunity in the general Egyptian population. The surveyed population consisted of 709 healthy subjects from five regions of Egypt, 2 month to 105 years of age. We used an ELISA assay to determine titers of anti-tetanus IgG. According to the widely used criteria, 31.7% was susceptible to tetanus (IgG level < 0.15 IU/ml), 15.7% protection (0.15-1.0 IU/ml), and 52.6% > or = 1.0 IU/ml). The results revealed that the majority (68.3%) population from 2 months to 50 years had a protective level of IgG against tetanus. The level of susceptibility increases with age until it reaches 90.3% Significantly, more males (23.7%) unprotected against tetanus. Our results suggest that monitoring immunization status and administering the tetanus vaccine as required are essential to insure adequate and long-lasting antibody levels.
The Egyptian journal of immunology / Egyptian Association of Immunologists, 2004
The aim of this study was to determine immune status to corynebacterium diphtheria by screening f... more The aim of this study was to determine immune status to corynebacterium diphtheria by screening for protective antibodies in a sample of Egyptian population. The study population consisted of 709 healthy subjects aged from 2 months to 105 years, inhabitants of 6 regions of Egypt. The study utilized Enzyme linked immunosorbent assay (ELISA) to measure serum levels IgG antibodies reactive with diphtheria toxoid. Levels of diphtheria toxoid antibody > or = 0.1 IU/ ml were defined as immune/protected, 23.9 % of the population were found to be susceptible to diphtheria (IgG level < 0.01 IU/ml), 43% had basic protection (0.01-0.09 IU/ml), and 33.1% were fully protected (0.1 IU/ml). The results revealed that serum levels of antitoxin antibodies decreased in old ages (< 60 y) with the females being more susceptible then males. These results recommend a booster immunization for the susceptible age groups.
Testosterone and 5alpha-dihydrotestosterone (DHT) are the principal male hormones (androgens) in ... more Testosterone and 5alpha-dihydrotestosterone (DHT) are the principal male hormones (androgens) in mammals. The enzyme, steroid 5alpha reductase catalyzes the conversion of testosterone (T) to its biologically potent steroid (DHT) in androgen dependent tissues. Two 5alpha reductase isoenzymes have been identified in rat tissues. The type I isoenzyme has been shown to be predominately expressed in the rat liver, whereas androgen target tissues of the genital tract express mainly isoenzyme II. The effects of androgens and glucocorticoids on the abundance of steroid 5alpha reductase type I (5alphaR1) messenger RNA in the rat liver were examined. Steady state levels of 5alphaR1 mRNA decreased dramatically to 1.5% of control levels 14 days following adrenalectomy (ADX), whereas dexamethasone (Dex) administered (0.5 mg/100 g) to ADX animals enhanced the expression of 5alphaR1 to twice its' normal values within 40 hours. Bilateral orchiectomy induced, within eight days, the expression of...
A case of familial Leydig cell hypoplasia as a cause of male pseudohermaphroditism is described i... more A case of familial Leydig cell hypoplasia as a cause of male pseudohermaphroditism is described in two 46,XY female sibs. Biochemical and histologic evidence for such diagnosis is presented.
In an attempt to evaluate the levels of immunity against diphtheria in the Egyptian population, 7... more In an attempt to evaluate the levels of immunity against diphtheria in the Egyptian population, 709 healthy subjects aged 2 months-105 years and from six regions of Egypt were investigated. An ELISA was used to determine the serum concentration of anti-diphtheria IgG in each subject. Following widely-used criteria for defining levels of protection, 34.1% of the subjects were categorized as susceptible to diphtheria (with &lt; 0.01 IU antitoxin/ml serum), 43.7% were considered to have basic protection (0.01- &lt; 0.1 IU/ml) and only the other 22.1% appeared fully protected (&gt; or = 0.1 IU/ml). The results revealed that most of the subjects aged 2 months-50 years had a basic or fully protective level of IgG against diphtheria, although males were slightly more likely to be unprotected than females (36.2% v. 31.6%) and certain age-groups appeared to be much more likely to be susceptible than others. If outbreaks of diphtheria like those seen in recent years in the former Soviet Union are to be avoided in Egypt, the most susceptible groups of the population need to be given booster immunizations.
Enzyme kinetics of 5 alpha-reductase were compared in cultured genital skin fibroblasts taken fro... more Enzyme kinetics of 5 alpha-reductase were compared in cultured genital skin fibroblasts taken from 6 control subjects and from an affected subject with 5 alpha-reductase deficiency in whom the diagnosis was established on hormonal grounds. The Km value for testosterone in the mutant enzyme was extremely high (1,427 vs. 185-417 nmol/l in controls). in the mutant enzyme was extremely high (1,427 vs 185-417 nmol/l in controls). A A mutant but stable enzyme with reduced affinity to steroid substrate is reported.
Two transgenic mice were produced by microinjection of the entire hepatitis B virus (HBV) genome ... more Two transgenic mice were produced by microinjection of the entire hepatitis B virus (HBV) genome as a 3.2-kilobase EcoRI DNA fragment into one-cell embryos. Each animal contained a single, unique locus of HBV sequence. One founder animal, G7, contained a partially deleted HBV genome lacking both putative HBV surface antigen (HBsAg) promoters. The other animal, G26, contained greater-than-genome-length HBV sequences organized as a partial head-to-tail dimer. Both transgenic animals transmitted the HBV sequences in a Mendelian fashion, and all subsequent transgenic animals had detectable HBsAg in the serum. Expression of HBV sequences in tissues from G7- and G26-derived mice showed preferential expression of the 2.1-kilobase HBsAg RNA transcript in liver and kidney tissues by Northern (RNA) blot analysis. These data are consistent with the notion that HBV DNA contains cis-acting regulatory sequences which are responsible for the predominant expression of HBsAg transcripts in the liver...
Molecular genetic analysis of individuals from 6 Egyptian and 33 German families with fragile X s... more Molecular genetic analysis of individuals from 6 Egyptian and 33 German families with fragile X syndrome and 240 further patients with mental retardation was performed applying a completely non-radioactive system. The aim of our study was the development of a non-radioactive detection method and its implementation in molecular diagnosis of the fragile X syndrome. Furthermore, we wanted to assess differences in the mutation sizes between Egyptian and German patients and between Egyptian and German carriers of a premutation. Using non-radioactive polymerase chain reaction (PCR), agarose gel electrophoresis and blotting of the PCR products, followed by hybridisation with a digoxigenin-labelled oligonucleotide probe (CGG)5 and chemiluminescent detection, we identified the fragile X full mutation (amplification of a CGG repeat in the FMR-1 gene ranging from several hundred to several thousand repeat units) in all patients. We observed no differences in the length of the CGG repeat between the Egyptian and German patients and carriers, respectively. However, in one prenatal diagnosis, we detected only one normal sized allele in a female fetus using the PCR-agarose assay, whereas Southern blot analysis with the digoxigenin labelled probe StB 12.3 revealed presence of a full mutation. Our newly established nonradioactive genomic blotting method is based on the conventional radioactive Southern blot analysis. Labelling of the probe StB 12.3 with digoxigenin via PCR allowed the detection of normal, premutated and fully mutated alleles. For exact sizing of small premutated or large normal alleles, we separated digoxigenin labelled PCR products through denaturing polyacrylamide gelelectrophoresis (PAGE) and transfered them to a nylon membrane using a gel dryer. The blotted PCR-fragments can easily be detected with alkaline phosphate-labelled anti-digoxigenin antibody. The number of trinucleotide repeat units can be determined by scoring the detected bands against a digoxigenated M13 sequencing ladder. Our newly developed digoxigenin/chemiluminescence approach using PCR and Southern blot analysis provides reliable results for routine detection of full fragile X mutations and premutations.
Egyptian Journal of Medical Human Genetics, May 1, 2002
... Assessment of the Pattern of Cerebral Blood Flow in Patients with Fragile X Syndrome Using Si... more ... Assessment of the Pattern of Cerebral Blood Flow in Patients with Fragile X Syndrome Using Single Photon Emission Computed Tomography Hanan H. Afifi,1 Alice K. Abd EI-Aleem,2 Laila F. Malta,3 Adel M. Ashour,1 Mostafa K. EI-Awady,2 Nagwa A. Meguid1. ...
Eastern Mediterranean health journal = La revue de santé de la Méditerranée orientale = al-Majallah al-ṣiḥḥīyah li-sharq al-mutawassiṭ
Analysis of expressed mRNAs with differential display-polymerase chain reaction (DD-PCR) is a pow... more Analysis of expressed mRNAs with differential display-polymerase chain reaction (DD-PCR) is a powerful tool for the characterization of genes involved in malignant pathways and might identify markers for different phases of chronic myelogenous leukaemia (CML). We examined the presence of BCR-ABL transcripts in 25 CML patients in either the chronic phase or blast crisis. We then analysed the expression of leukocytic RNA transcripts in CML phases. DD-PCR technique was used to examine CML cases with BCR-ABL in comparison with CML cases lacking detectable BCR-ABL transcripts. Our results support the use of differential display not only for characterization of the CML differentially expressed genes but also to locate patterns that can be implemented as valuable fingerprints for each phase of CML.
Nature and Science, 6(2), 2008, ISSN: 1545-0740, naturesciencej@gmail.com ... CTT Multiplex Syste... more Nature and Science, 6(2), 2008, ISSN: 1545-0740, naturesciencej@gmail.com ... CTT Multiplex System is a quick and inexpensive method to exclude innocent suspects during criminal inquiries ... Ahmed M. Refaat1, Wael T. El-garf2, Kholoud S. Ramadan3, Mohamed M. ...
Our aim was to determine the level of tetanus immunity in the general Egyptian population. The su... more Our aim was to determine the level of tetanus immunity in the general Egyptian population. The surveyed population consisted of 709 healthy subjects from five regions of Egypt, 2 month to 105 years of age. We used an ELISA assay to determine titers of anti-tetanus IgG. According to the widely used criteria, 31.7% was susceptible to tetanus (IgG level &lt; 0.15 IU/ml), 15.7% protection (0.15-1.0 IU/ml), and 52.6% &gt; or = 1.0 IU/ml). The results revealed that the majority (68.3%) population from 2 months to 50 years had a protective level of IgG against tetanus. The level of susceptibility increases with age until it reaches 90.3% Significantly, more males (23.7%) unprotected against tetanus. Our results suggest that monitoring immunization status and administering the tetanus vaccine as required are essential to insure adequate and long-lasting antibody levels.
The Egyptian journal of immunology / Egyptian Association of Immunologists, 2004
The aim of this study was to determine immune status to corynebacterium diphtheria by screening f... more The aim of this study was to determine immune status to corynebacterium diphtheria by screening for protective antibodies in a sample of Egyptian population. The study population consisted of 709 healthy subjects aged from 2 months to 105 years, inhabitants of 6 regions of Egypt. The study utilized Enzyme linked immunosorbent assay (ELISA) to measure serum levels IgG antibodies reactive with diphtheria toxoid. Levels of diphtheria toxoid antibody > or = 0.1 IU/ ml were defined as immune/protected, 23.9 % of the population were found to be susceptible to diphtheria (IgG level < 0.01 IU/ml), 43% had basic protection (0.01-0.09 IU/ml), and 33.1% were fully protected (0.1 IU/ml). The results revealed that serum levels of antitoxin antibodies decreased in old ages (< 60 y) with the females being more susceptible then males. These results recommend a booster immunization for the susceptible age groups.
Testosterone and 5alpha-dihydrotestosterone (DHT) are the principal male hormones (androgens) in ... more Testosterone and 5alpha-dihydrotestosterone (DHT) are the principal male hormones (androgens) in mammals. The enzyme, steroid 5alpha reductase catalyzes the conversion of testosterone (T) to its biologically potent steroid (DHT) in androgen dependent tissues. Two 5alpha reductase isoenzymes have been identified in rat tissues. The type I isoenzyme has been shown to be predominately expressed in the rat liver, whereas androgen target tissues of the genital tract express mainly isoenzyme II. The effects of androgens and glucocorticoids on the abundance of steroid 5alpha reductase type I (5alphaR1) messenger RNA in the rat liver were examined. Steady state levels of 5alphaR1 mRNA decreased dramatically to 1.5% of control levels 14 days following adrenalectomy (ADX), whereas dexamethasone (Dex) administered (0.5 mg/100 g) to ADX animals enhanced the expression of 5alphaR1 to twice its' normal values within 40 hours. Bilateral orchiectomy induced, within eight days, the expression of...
A case of familial Leydig cell hypoplasia as a cause of male pseudohermaphroditism is described i... more A case of familial Leydig cell hypoplasia as a cause of male pseudohermaphroditism is described in two 46,XY female sibs. Biochemical and histologic evidence for such diagnosis is presented.
In an attempt to evaluate the levels of immunity against diphtheria in the Egyptian population, 7... more In an attempt to evaluate the levels of immunity against diphtheria in the Egyptian population, 709 healthy subjects aged 2 months-105 years and from six regions of Egypt were investigated. An ELISA was used to determine the serum concentration of anti-diphtheria IgG in each subject. Following widely-used criteria for defining levels of protection, 34.1% of the subjects were categorized as susceptible to diphtheria (with &lt; 0.01 IU antitoxin/ml serum), 43.7% were considered to have basic protection (0.01- &lt; 0.1 IU/ml) and only the other 22.1% appeared fully protected (&gt; or = 0.1 IU/ml). The results revealed that most of the subjects aged 2 months-50 years had a basic or fully protective level of IgG against diphtheria, although males were slightly more likely to be unprotected than females (36.2% v. 31.6%) and certain age-groups appeared to be much more likely to be susceptible than others. If outbreaks of diphtheria like those seen in recent years in the former Soviet Union are to be avoided in Egypt, the most susceptible groups of the population need to be given booster immunizations.
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