C.V Ashraf Hassan 2010...Clinical Biochemistry course ( Enzymes, metabolism and metabolic disorders ) for undergraduate students at Faculty of Health Sciences, Jazan University, Kingdom

C.V. ( Ashraf A. M. Hassan, Ph.D. )

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CURRICULUM VITAE

Appointments Sep. 2009 – Current Professor of Medical Biochemistry at Faculty of Health Sciences,

Jazan University, Kingdom Saudi Arabia

June 2006 - Sept. 2009 Professor of Medical Biochemistry, Medical Research Institute, Alexandria University, Egypt

Nov. 1997 – May 2006 Associate Professor of Medical Biochemistry, Medical Research Institute, Alexandria University, Egypt

Nov. 1995 – Oct. 1997 Lecturer of Medical Biochemistry, Medical Research Institute, Alexandria University, Egypt

Personal Information:

Name Ashraf Aly Moustafa HASSAN Title Professor of Medical Biochemistry Nationality Egyptian Date of Birth March 26, 1962 Place of Birth Alexandria, Egypt. Marital Status Married

Affiliation Professor of Medical Biochemistry and Molecular Biology, Medical

Laboratory Department, Faculty of Health Sciences, Jazan University, Kingdom of Saudi Arabia.

Contact Address Mobile

+966-562074888 / +966-541186428 / +20-178858828

E-mail [email protected] / [email protected]

Education 2006 Professor Title in Medical Biochemistry

1997 Associate Professor Title in Medical Biochemistry

1988 - 1991 Ph.D. in Medical Biochemistry, Alexandria University, Medical Research Institute in Channel Program Fellowship with Wakayama Medical College, Japan (Date of Award: 1991).

1983-1987 M.Sc. in Medical Biochemistry, Alexandria University, Medical Research Institute (Date of Award: 1987).

1979-1983 B.Sc. in Biochemistry, Alexandria University, Faculty of Sciences (Date of Award: 1983).

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Oct. 1993 – Oct. 1995 Postdoctoral Research Fellow in Molecular Toxicology department, Mario Negri Institute, Milan, Italy

March 1991 – Sept. 1993 Lecturer of Medical Biochemistry, Medical Research Institute, Alexandria University, Egypt

Dec. 1988 – Feb. 1991 Ph.D. Research Fellow in Medical Biochemistry Department, Wakayama Medical College, Wakayama, Japan

June 1987 – Dec. 1988 Assistant Lecturer in Applied Medical Chemistry Department, Medical Research Institute, Alexandria University, Egypt

Nov. 1983 –May 1987 Demonstrator in Applied Medical Chemistry Department, Medical Research Institute, Alexandria University, Egypt

Teaching Experience

Clinical Biochemistry course ( Enzymes, metabolism and metabolic disorders ) for undergraduate students at Faculty of Health Sciences, Jazan University, Kingdom Saudi Arabia.

Endocrinology & Hormones course for undergraduate students at Faculty of Health Sciences, Jazan University, Kingdom Saudi Arabia.

Clinical Biochemistry course ( Electrolytes heomostasis, Liver disease, Renal diseases and cardiac diseases ) for undergraduate students at Faculty of Health Sciences, Jazan University, Kingdom Saudi Arabia.

Basic Biochemistry for undergraduate and postgraduate students at Medical Research Institute

Biochemistry courses for Medical Laboratory Science undergraduate students, Faculty of Applied Medical Sciences, Pharos International University, Alexandria, Egypt

Applied Medical Chemistry Course for Master and Ph.D. degree students

Cancer Chemistry Course ( DNA damage and repair mechanisms, carcinogens and mechanism of carcinogenesis ) for Master and Ph.D. degree students

Molecular Biochemistry Course ( PCR theory and application in diagnosis )for Master and Ph.D. degree students

Medical Biochemistry Course ( Biochemistry of diseases, Biochemistry of nutrition )for Ph.D. students

Research Experience Basic biochemical techniques

Molecular biology basic laboratory techniques

DNA and RNA extraction and purification techniques

Electrophoresis methods for Protein, DNA and RNA





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Conventional PCR , RT-PCR, Real Time PCR

PCR applications in diagnosis

HPLC applications in Biochemistry and Environmental pollution studies

Gas Chromatography-Mass Spectrometry ( GC-MS ) applications in detection of carcinogenic metabolites and environmental pollutants.

Recent Research Work

Diagnosis of H. pylori infection and risk factor of the presence of BabA2 gene using PCR technique ( Ongoing research work )

A molecular approach for detection of the schistosomicidal effect of Mirazid (Myrrh) in the murine model ( In Press )

PCR Based Diagnosis of H. pylori Infection and Risk Factor of the Presence of iceA & ureC Genes ( Published as abstract 2010 )

Study of Gene Expression Level of p27 and MDM2 in Exfoliated Cells from Bladder Cancer Patients ( Published 2008 )

Expression of Cyclo-Oxygenase-2 (COX-2) in Patients with Hepatocellular Carcinoma ( Under Publication )

Accumulation of Hypoxia-Inducible Fctor-1 α (HIF-1α) Protein and Cyclooxygenase-2 (COX-2) Gene Expression in Human Breast Cancer Tissue ( Under Publication )

Molecular Diagnosis of H. pylori Infection and Risk Factor of the Presence of cagA & vagA Genes ( Under Publication )

Research Supervision

M. Sc. Thesis Main Supervisor for 6 theses and co-Supervisor for 4 theses in Applied Medical Chemistry Dept., Medical Research Institute, Alexandria University, Egypt

Ph.D. Thesis Co-Supervisor in 4 theses in Applied Medical Chemistry Dept., Medical Research Institute, Alexandria University, Egypt

Language Profeciency

Arabic Mother tongue

English Very good command in writing, speaking and reading

Conferences & Meetings Attended

Actively participated in the 14th International Conference of International Society of Infectious Diseases ( ISID ) held in Miami, Florida, USA ( 9 – 12 March, 2010 ).

Presented 2 posters.





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Professional Membership

International Study Group of tryptophan Research ( ISTRY )

International Society of Infectious Diseases ( ISID )

Arab Society of Biotechnology

Japanese Study Group of tryptophan Research ( JSTRY )

Egyptian Association of Medical Biochemistry and Molecular Biology

Scientific committee at Medical Research Institute, Alexandria University

Member of the Institutional Review Board ( IRB ) of the Jazan Medical Research Center ( JMRC ), Jazan University, Kingdome Saudi Arabia

Member of the Deanship of Scientific Research Committee, Jazan University, Kingdome Saudi Arabia

Other Academic Activities Active participant and Facilitator at the Intensive Training Course in Research

Methodology for Health Sciences Students held the Medical Research Center of Jazan University, KSA ( 17- 18 & 24 - 25 March, 2010 )

Established the Molecular Biochemistry Laboratory at Applied Medical Chemistry Department, Medical Research Institute, Alexandria University, Egypt

Organizer of training workshop on basic techniques in molecular biology held at Applied Medical Chemistry Department, Medical Research Institute, Alexandria University, Egypt ( 26 – 28 August, 2008 )

One of 3 members committee established the Faculty of Allied Medical Science, Pharos International University, Alexandria, Egypt ( 2009 ).

Established the Medical Laboratory Science Department at the Faculty of Allied Medical Science, Pharos International University, Alexandria, Egypt ( 2009 ).

Alexandria University Award for Scientific Encouragement, 1997

Current Research Projects

Partner of the project entitled " Development of Nanomaterials for Drug Delivery Systems, Sensors, and Photovoltaic Applications " funded by Research Enhancement Program ( ALEX REP ), 2008

Scientific Publications

1- Lotfy WM, Nageh AM, Hussein NAM, Hassan AAM. A molecular approach for detection of the schistosomicidal effect of Mirazid (Myrrh) in the murine model. Eastern Mediterranean Health Journal ( In Press )

2- Hassan A et al. A diagnostic approach to detect murine Schistosoma mansoni infection using polymerase chain reaction. ( Accepted Poster ) International Society for Infectious Disease (ISID) Conference, FL, USA 2010.





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3- Hassan A et al. Molecular diagnosis of Helicobacter pylori infection and risk factor of the presence of cagA and vacA genes. ( Accepted Poster ) International Society for Infectious Disease (ISID) Conference, FL, USA 2010.

4- Hassan A et al. Molecular Diagnosis of Helicobacter pylori Infection and Risk Factor of the Presence of cagA and vacA Genes ( Accepted Poster ) Com Bio Conference, New Zealand 2009.

5- El-Abd E, Hassan A., El-Ashry O, El-Abshity M, El-Sewedy SM. P27 and mdm2 as molecular grading biomarkers in transitional cell carcinoma. Turkish J Cancer 2008; 38(2): 68-77.

6- Mostafa AA, Hassan AAM, Hassan A. Low Flow sevoflurane anesthesia and its acute impact on renal biochemical markers. Bull Egypt Soc Physiol Sci 2005; 25(2): 329

7- Hassan AA, Zein El-Din M, Fawzy A, Mokhtar M. Molecular study on methylenetetrahydrofolate reductase (MTHFR) and factor V leiden (FV) genes polymorphism as risk factor for preeclampsia in Alexandria, Egypt.

Bull Egypt Soc Physiol Sci 2005; 25(2): 317– 328 8- Ismail SS, Ebeid SA, Hassan AA, Abdel-Mohsen MA, Desouki A. Effect of repeated

exposure to N-nitrosodimethylamine (NDMA) on oxidative stress in S. mansoni infected mice. Bull Egypt Soc Physiol Sci 2005; 25(2): 247– 260

9- Hassan EM, Hassan AA, Hamdy HA, Aboul Azm T, El-Sewedy SM. Glutathione S-transferase Mu (GST-µ) isoenzyme as a possible biomarker in the development of bladder carcinoma in schistosoma- infected patients.

Bull Egypt Soc Physiol Sci 2005; 25(2): 145– 158 10- Hassan AA, El-Sherbini E, El-Sawaf G, Hamdy H, El-Toukhy ZA. Biochemical view

on the effect of hepatitis delta virus infection on expression of hepatitis B virus in blood donors. Bull High Inst Pub Health 2005; 35(1): 127-142

11- Mostafa AA, Hassan AA. The efficacy of dexmedetomidine in suppressing cardiovascular and sympathoadrenal response to endotracheal intubation in hypertensive patients undergoing elective surgery. Bull High Inst Pub Health 2004; 34(4): 985-994

12- Hassan AA, Amin GA, Abdel-Mohsen MA, Thabet R, Fadaly GA, El-Sewedy SM. Role of oxidative stress, GST-pi isoenzyme and p53 gene mutation in development of hepatocellular carcinoma in patients with chronic viral hepatitis C with and without schistosomiasis. J Med Res Inst 2004; 25(6): 152-178

13- Hassan AA, Youssef AI, Ibrahim SA, El-Khouly ZA. Evaluation of the role of Schistosoma mansoni infection on renal mixed function oxidases (MFO) in DMN injected mice. J Med Res Inst 2002; 23(2): 191-203

14- Morsi MI, Abdel-Moneim NA, Hassan AA, Al-Sharaky AS. Radioimmunoasssay of plasma glutathione S-transferase-pi as a tumour marker in hepatocellular carcinoma.

Bull Alex Fac Med 2001; XXXVII (3): 379-384





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15- Abdel-Mohsen MA, Hassan AA, El-Sewedy SM, Aboul-Azm T, Mgagnotti C, Fanelli R, Airoldi L. Human bladder cancer, schistosomiasis, N-nitroso compounds and their precursors. Int J Cancer 2000; 88(4): 682-3

16- Abdel-Mohsen MA, Hassan AA, El-Sewedy SM, Aboul-Azm T, Mgagnotti C, Fanelli R, Airoldi L. Biomonitoring of N-nitroso compounds, nitrite and nitrate in the urine of Egyptian bladder cancer patients with and without Schistosoma haematobium infection.

Int J Cancer 1999; 82(6): 789-94 17- Hassan AA, Tagliabue G, Codegoni AM, D`Incalci M, El-Sewedy SM, Airoldi L.

Glutathione S-transferase activity and glutathione content in human bladder carcinoma associated with schistosomiasis: comparison with uninvolved surrounding tissues.

Cancer Letters 1997; 121: 19-23. 18- Hassan AAM, Airoldi L, El-Sewedy SM. Glutathione content and glutathione S-

transferase activity in urinary bladder tumor tissues associated with schistosomiasis.

J Med Res Inst 1997; 18(1): 90-97 19- Hassan AA. Effect of intraperitoneal subleathal doses of dimethoate on rat hepatic

gluconeogenesis. J Med Res Inst 1997; 18(1): 117-123.

20- Hassan AAM, Benfenati E, Facchini G, Fanelli R. Pollution of ground and drinking water with volatile organic compounds: solid-phase microextraction and GC/MS analysis. Toxicol Environ Chem 1996; 55: 73-81

21- Hassan AA, Benfenati E, Fanelli R. detection and quantification of trihalomethanes in drinking water from Alexandria, Egypt.

Bull Environ Contam Toxicol 1996; 56(3): 397-404 22- Hirai T, Minatogawa Y, Hassan AAM, Kido R. Metabolic inter-organ relations by

exercise of fed rat: carbohydrates, ketone body and nitrogen compounds in splanchnic vessels. Physiology & behavior 1995; 57(3): 515-522.

23- Hassan AAM, El-Khalili MM, Hussein NG, Kido R. Changes in serum lipid profile and esterases of rats after sublethal daily doses of dimethoate.

J Egypt Pub Health Ass. 1995; 70(3,4): 432-447. 24- Hassan AAM. Inhibition of gluconeogenic enzymes in rat liver after intraperitoneal

sublethal doses of dimethoate. Bull High Inst Pub Health 1995; 25(3): 699-704.

25- Hassan AA, Minatogawa Y, Hirai T, Kido R. Changes of some serum parameters and amino acids contents in rats after chronic sublethal doses of dimethoate.

Arch Environ Contam Toxicol 1994; 27(2): 256-9 26- Hassan AAM. Malnutrition diabetes and concomitant pancreatic changes (an

experimental study). Egypt J Med Lab Sci 1994; 3





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27- El-Sharkawy AM, Abdel-Rahman SZ, Hassan AA, Gabr MH, El-Zoghby SM, El-Sewedy SM. Biochemical effects of some insecticides on the metabolic enzymes regulating glutathione metabolism.

Bull Environ Contam Toxicol 1994; 52(4): 505-10

28- El-Khalili MM, Helmy MH, Hassan AAM, Abul-Azm T, El-Sewedy SM. Studies on urinary alpha-naphthyl acetate esterase isoenzymes in bladder cancer patients.

Egyp J Med Sci 1993; 14(1): 195-205

29- Hassan AAM, El-Khalili M, Abdel-Rahman SZ, El-Sewedy SM. Effect of gamma radiation on tryptophan metabolism in rat liver and kidney.

Advanced Tryptophan Research (Japan) 1992; 167-170.

30- Hassan AA, El-Sewedy SM, Minatogawa Y, Kido R. In vitro effect of dimethoate on the activity of tryptophan pyrrolase in rat liver. J Environ Sci Health 1991; 26(3): 333

31- Hassan AA, El-Sewedy SM, Minatogawa Y, Kido R. Effect of some insecticides on several enzymes of tryptophan metabolism in rats.

J Environ Sci Health 1990; 25(3): 333-46 32- El-Toukhy MA, Ebied SA, Hassan AA, El-Sewedy SM. In vivo studies on the effect of

some insecticides on the hepatic activities of L-tryptophan 2,3-dioxygenase and pyridoxal phosphokinase of male mice. J Environ Sci Health B. 1989; 24(3): 265-76

Applicant Declaration

I hereby certify that, the information given above is true, correct and accurate, that describes myself, my qualifications and experience to the best of my knowledge and belief. Applicant's signature --------------------------------------------

Updated: 13th August, 2010





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Abstracts of Publications

( 20 abstracts ) Dr. Ashraf Aly Moustafa Hassan

Faculty of Health Sciences, Jazan University





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Eastern Mediterranean Health Journal ( In Press )

A molecular approach for detection of the schistosomicidal effect of Mirazid® (myrrh) in the murine model.

Ashraf A.M. HassanWael M. Lotfy, Aly M. Nageh, Neveen A.M. Hussein,

ABSTRACT We used the conventional PCR technique for studying the schistosomicidal effect of

Mirazid® in the murine model, and comparing the results of the molecular study with

the parasitological results (ova and worm count). The used PCR technique was more

sensitive than the Kato-Katz thick smears. Mirazid® showed some schistosomicidal

effects against murine S. mansoni. However, it was not efficient enough to cure any

of the studied mice.

International Society for Infectious Disease (ISID) Conference, Miami, FL, USA (2010) Poster Presentation

A diagnostic approach to detect murine S. mansoni infection using a polymerase chain reaction.

**Mohammed AreeshiNeveen Hussein, Ali Nageh, , *, Wael Lotfy ssanAshraf Ha

ABSTRACT We used the conventional PCR technique for studying the schistosomicidal effect of

Mirazid® in the murine model, and comparing the results of the molecular study with

the parasitological results (ova and worm count). The used PCR technique was more

sensitive than the Kato-Katz thick smears. Mirazid® showed some schistosomicidal

effects against murine Shistosoma mansoni. The Kato-Katz technique showed false

negatives in 80% of the Mirazid® treated mice. However, the PCR gave lower

percentage of false negatives (40%) in the treated group. This may indicate that the

qualitative PCR technique is more sensitive than the Kato-Katz thick smears.

International Society for Infectious Disease (ISID) Conference, Miami, FL, USA (2010) Poster Presentation

Molecular Diagnosis of Helicobacter pylori Infection and Risk Factor of the Presence of cagA and vacA Genes.





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Saleh Aal, M. Sheta, -Afand, S. Abdel-, N. Dwidar, G. Hilaly, N. El*Ashraf Hassan**lahAbdul

ABSTRACT The aim of our study was to evaluate the efficiency of using PCR technique as a

powerful tool besides being an easy and low cost method for diagnosis of H pylori

infection through molecular detection of cagA and vacAs1 genes in gastric tissue

biopsies obtained from patients diagnosed as H pylori positive. There were 50 cases

enrolled in our study that underwent endoscopy because of clinical symptoms

attributable to upper gastrointestinal tract. The following investigations were done:

Routine upper endoscopy, Histological examination to grade the severity of gastritis

to detect H. pylori as a gold standard, Biochemical analysis for all cases were carried

on gastric biopsies, included CLO-Rapid urease test and PCR analysis for detection

of cagA and vacAs1 genes in gastric tissue biopsies obtained from patients

diagnosed as H pylori positive.

The histological results of this study revealed that: Out of 50 patients only 41 was H.

pylori positive. 9(18%) patients showed grade 0 infection on histological examination,

while the remaining 41 (82%) patients showed different grades of infection, ranging

from grade 1 to grade2, where the highest percentage of patients (62%) showed

grade1.

The obtained biochemical results of this study revealed that: Out 41 positive cases

by histology, only 36 (87.80%) were also positive by the rapid urease test. On the

other hand, out of 9 that were negative by histology 8 (88.89%) were negative by

rapid urease test.

The PCR results of this study revealed that: Out of 41 positive cases by histology,

only 40 (97.56%) were also positive by the PCR test for vacAs1 gene. On the other

hand, out of 10 that were negative by PCR test for vacAs1 gene 9 (100.00%) were

negative by histology.

The correlation results of this study revealed that: Strong positive and statistically

significant correlation between the expressions of these two genes (vacAs1 and

cagA) in patient of H. pylori. (P< .000)

Turkish Journal of Cancer: 38(2), 68-77 (2008)

P27 and mdm2 as molecular grading biomarkers in transitional cell carcinoma.

-Ipshiti, Shehata El-Ashry, Magda Al-, Osama ElassanH Ashraf, Abd-Eman El

Sewedy





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ABSTRACT To investigate the clinical relevance of p27 and mdm2 gene expression in transitional cell carcinoma (TCC), urine samples were collected from 30 TCC

patients (preoperatively), 10 patients with nonmalignant urological diseases and 10 controls. Total RNA was extracted from urine sediments and RT-PCR was performed. Absence of p27 mRNA

and mdm2 acquisition was specific for grade III TCC. Recurrences significantly increased with schistosomal infection and larger tumor size. Overall survival significantly

shortened with larger tumor size, tumor multiplicity, and solid tumor growth pattern. In multivariate analysis, the tumor growth pattern was significant prognostic factor (p=0.036) for disease-free and overall survival while, high grade (exactly significant) and solid tumor growth pattern (p=0.023) were significant prognostic factors for the p27-/mdm2+ genetic pattern in TCC. We conclude that efficient management of TCC

would be enhanced using p27 or mdm2 mRNAs as molecular grading markers.

Bull High Institute Public Health: 35 (1), 127 – 42 ( 2005)

Biochemical View on the Effect of Hepatitis Delta Virus Infection on Expression of Hepatitis B Virus in Blood Donors.

-Sawaf, Hanan Hamdy, Zienat El-Sherbini, Gamal El-, Eglal ElAshraf A HassanKholy

ABSTRACT One thousand voluntary blood donors attending the Blood Bank of Medical Research Institute, Alexandria University were enrolled in the present study. Sera from blood samples were separetd for screening of HBsAg using ELISA technique. Screening of 200 HBsAg negative blood samples for past exposure to HBV infection was also performed through the detection of anti-HBc using specific ELISA kit. Determination of ALT and AST revealed a significant increase among HBsAg positive group while, Malondialdehyde (MDA) detection revealed the absence of the causative relationship between MDA and liver injury. On the other hand, no anti-HDV could be found among the HBsAg –ve/anti-HBc +ve group of blood donors (46 cases). PCR analysis for HBV-DNA was performed in the chosen 24 serum samples. None of them was positive for HBV-DNA regardless their positively or negativity to the different

serologic markers studied.

Bull Egypt Soc Physiol Sci : 25 (2), 317 – 28 (2005)

Molecular Study on Methylenetetrahydrofolate Reductase (MTHFR) and Factor V Lieden (FV) Genes Polymorphisms as risk factor for Preeclampsia in

Alexandria, Egypt

tarEldin, A Fawzy, M Mokh-, M ZeinAA Hassan





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ABSTRACT

The main objective of the present study is to investigate the potential effects of Factor V leiden (FV) mutation and 5,10 methylenetetrahydrofolate reductase C677T

(MTHFR) polymorphosim in preeclampsia women from Alexandria, Egypt.

A total of 93 preeclampsia women and 73 healthy pregnant women controls were recruited and evaluated for frequency of Leiden and MTHER mutations. Genomic DNA was isolated from a venous blood samples and then genotyped by using polymerase chain reaction Restriction Fragment Length Polymorphism (PCR-RFLP).

Detection of C677T MTHFR gene polymorphism was done using HinfI restriction endonucleasse while G1691A Factor V lieden using MnlI enzyme. An optimized PCR cycling program was set in laboratory as follows: initial denaturation at 95°C for 5 min, followed by 37 cycles of denaturation at 95°C for 30 s, annealing at 54°C for 45 s (at 59°C for 30 s for Factor V leiden), elongation at 72°C for 45 s and a final elongation step at 72°C for 5 min. The final product was run on 2% and 3.5%

agarose and is visualized after staining with ethidium bromide.

Laboratory analysis yield 31% heterozygous carriers of FV mutation among severe preeclampsia women and 7.8% heterozygous carriers among the healthy controls; differences between the two groups were found to be statistically significant (OR = 5.32, CI: 1.50 – 19.5). Heterozygous MTHFR mutations were found in 39 (41.9%) preclamptic women and in 31 (40.3%) control group. MTHFR gene polymorphism did not appear to be correlated with the development of preeclampsia (OR= 1.05; CI: 0.4 – 2.72). The frequency of mutation in combination (MTHFR and FV) was significantly increased in preeclampsia patients compared to control group (OR=

3.14; CI: 1 – 23.3).

We can conclude from our present study that leiden mutations and the combination of two polymorphism may play a considerable role for women with preeclampsia. Meanwhile, a larger scale study is required to investigate the role of these two

parameters as a risk factor for preeclampsia.

Bull Egyptian Soc Physiol

Sciences: 25 (2), 145 – 58 (2005)

Glutathione S-Transferase MU (GST-μ) Isoenzyme as a Possible Biomarker in the Development of Bladder Carcinoma in Schistosoma-infected Patients.

Sewedy-, S M ElAzm-, H A Hamdy, T AboulA A Hassan, E M Hassan

ABSTRACT

In the present work the relationship between glutathione S-transferase Mu (GST-µ) isoenzyme phenotype and the development of bladder cancer (BC) among smoking

and non-smoking patients infected with S. hematobium was studied. Blood samples were taken from 40 healthy control male subjects and 120 male patients infected with S. hematobium (60 without BC and 60 with BC) for detection of





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GST-µ. isoenzyme by ELISA technique. Serologically, all cases were positive for anti-schistosoma antibodies, which indicates a recent or an old schistosomal infection. Histopathological examination was performed for biopsies taken from the

bladder to confirm the diagnosis of malignancy. Heparinized blood samples were withdrawn to perform the assay of GST-µ isoenzyme using ELISA Mu kit which is highly specific for GST-µ and does not cross react with other GST isoenzymes or any other blood components. Individuals with

enzyme levels below 10 ng/ml of blood are considered to be deficient in GST-µ . In normal control subjects, the proportion of GST-µ. deficiency (47.5%) was comparable to that in patients without BC (48.3%). However, the proportion in patients with BC (78.3%) exceeded that of 48.3% in patients without BC with an odds ratio = 3.86 (95% confidence interval (CI) = 1.63-9.3; χ2 11.63, P<0.001). On the other hand, among smoking patients a significant higher increased risk of BC

was associated with GST-µ deficiency than in non-smokers (P=0.007 Vs P=0.04). In conclusion, the present findings could indicate that, GST-µ. deficiency could be a risk factor for the development of BC among patients infected with S. hematobium and smoking increased this risk. In addition, this isoenzyme can be used as a genetic biomarker for the susceptibility to Schistosma-induced BC. Thus it is recommended that, patients infected with S. hematobium should be screened for GST-µ and those who are negative for this isoenzyme especially smokers should be monitored for bladder cancer development. This policy could help early detection of

this type of cancer with subsequent better management and prognosis.

Bull Egypt Soc Physiol Sci: 25(2), 247 – 60 (2005)

Effect of repeated exposure to N-nitrosodimethylamine (NDMA) on oxidative stress in S. mansoni infected mice

A Desouki sen, Abdel Moh , MA Hassan, S Ismail, S Ebeid ABSTRACT

Both S. mansoni infection and nitrosodimethylamine (NDMA) are widely available in the environment and are able to increase the generation of free radicals in the livers of exposed animals. Furthermore, the wide availability of NDMA in the environment makes repeated exposure to low dose more likely to occur than single exposure to a large dose. Therefore, in the present study, we investigated the effect of S. mansoni infection (50 cercariae per mouse) and repeated injections of NDMA (7.5 mg/kg body weight) at weekly intervals on the oxidative stress in mouse liver. The study was done using 250 Swiss albino mice divided into 4 groups: (1) 50 normal controls, (2) 50 NDMA treated group, (3) 50 S. mansoni infected group and (4) 100 NDMA-treated and S. mansoni infected group. Seven mice from each group were sacrificed by cervical dislocation at time intervals 5, 7, 9, 11, 13 and 15 weeks post infection. Livers were exscised and washed in ice-cold saline and homogenized in 0.2 M phosphate buffer, pH 7.4. Hepatic malondialdehyde (MDA) level, superoxide dismutase (SOD) activity, vitamine E concentration and total protein content were

measured in all samples.

Results of the present study showed that MDA was high in the three studied groups throughout the study period indicating the presence of oxidative stress. Vitamin E concentration was high at the 5th week after S. mansoni infection and NDMA





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injection raising the possibility that it could be mobilized from other parts of the body to areas of oxidative stress. Thereafter, vitamin E concentration was reduced probably due to exhaustion of its stores in the body. The infection process lowered SOD activity throughout the study period. However, in the NDMA injected group,

SOD activity was initially increased at the fifth week followed by a decrease.

The measured hepatic vitamin E concentration in the control as well as in the different tested groups was high at the 5th week. At subsequent time points, there was a decrease without a definit pattern of changes with time. No significant differences were found between the levels of hepatic vitamin E in mice infected with S. mansoni and injected with NDMA and in counter part mice with either S. mansoni

infection or NDMA-injection.

In conclusion, S. mansoni infection, NDMA injection or a combination of both increased the MDA from the 5th to the 15th week of exposure. Antioxidant defences as vitamine E increased initially at the 5th week in the three studied groups and decreased thereafter suggesting the potential benefit of vitamin E administration to patients under stressful conditions. In the NDMA injected mice, SOD activity increased at the 5th week and decreased thereafter. In both other groups, reduction of SOD activity was found from start. Therefore, exposure of S. mansoni infected

mice to NDMA yield a similar effects to that of S. mansoni infection alone. J Med Res Inst 25: 152-78 (2004)

Role of oxidative stress, GST-pi isoenzyme and p53 gene mutation in development of hepatocellular carcinoma in patients with chronic viral

hepatitis C with and without schistosomiasis. Geylan A Mohsen,-, Gamal A M Amin, Mohamed A AbdelAshraf A M Hassan

Fadaly, Rashad Thabet, Shehata M El-Sewedy

ABSTRACT The present work was aimed to study the etiopathological factors for the development of hepatocellular carcinoma (HCC) in patients with chronic viral hepatitis C with and without schistosomiasis by evaluating the detoxification capacity of serum glutathione (GSH) and its related enzymes, plasma glutathione S-transferase-pi isoenzyme (GST-pi) as well as gene p53 mutation state in liver biopsies. Three groups of patients were included in this study, non-schistosomal-HCV group (15 patients), schistosoma-HCV group (20 patients) and HCC-HCV

group (11 patients) in addition to 13 healthy controls. To approach the aim of the present study, liver function tests and virological markers were carried out for all groups. Biochemical parameters including GSH content, GGT, GSH-RD, GPx and GST as well as GST-pi were also measured . Alteration in p53 gene expression was detected using immunohistochemistry technique for liver

biopsies. Results indicated a significant decrease in GSH-RD in non-schistosomal HCV and HCC groups as compared to control group while the serum level of GST and GST-pi were significantly increased in the 3 studied groups of patients when compared to controls. According to histopasthological study of HCV patients, the p53 protein expression has been found in 13.3% of cases in chronic mild active hepatitis, 40% in chronic active hepatitis and 72.3% in HCC patients. In patients with positive p53 expression, the serum AST and plasma GST-pi levels were significantly higher when

compared to patients with negative p53 expression.





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From this present study, it could be concluded that the modulations of GSH-detoxifying enzyme system occurs due to HCV infection either associated or not with schistosomal infection. These modulations lead to depletion in GSH level, decrease in both GSH-RD and GPx activity and increase in the GGT, GST and GST-pi level. This in turn affects the oxidant/antioxidant balance in favour of the increased oxidative stress. Unneutralized free radicals will react with cellular macromolecules, eg. DNA causing mutations in oncogens as p53 tumor supressor gene which is represented by its overexpression. Accordingly, this could lead to enhance the susceptibility of the biotransformation of normal hepatocytes into malignant one and in addition, GST-pi level could be used as indication for the silent transformation of

hepatocytes from normal cells into malignant one.

J Med Res Inst: 23 (2), 191-203 (2002)

Evaluation of the role of Schistosoma mansoni infection on renal mixed function oxidases (MFO) in DMN injected mice.

Kholy-m, Zeinat A El, Amany I Yousef, Shereen A IbrahiAshraf A Hassan

ABSTRACT

Four hundred male Swiss albino mice were used as the experimental animal model.

Batches of these mice were randomly selected and divided into four groups (control,

S. mansoni infected, dimethylnitrosamine injected and combined infected-

injected). Animals were sarcificed at time intervals: 24 hours, 2, 4, 6, 8, 10 and 12

weeks post-injection taking well matched members of each group. Cytochrome P450

level and cytochrome c-reductase activity were determined in mitochondrial free

supernatent of kidley homogenate and were serially monitored along time intervals.

Cytochrome P-450 contents of the injected groups, was fluctuating between non-

significant decrease and increase when compared to the corresponsing controls,

except at the 8th week, there was a significant decrease approximately 30% below its

control, which gradually recovered reaching its highest significant increase by

approximately 50% at vthe 12th week post treatment. On the other hand, combined

group showed two significant decreases in its cytochrome P-450 level, at 4th and 10th

weeks by approximately 18% and 34%, respectively compared to their

corresponding controls. In schistosomiasis infected group, cytochrome P-450 level

did not show any significant change from their corresponding control during the

whole period of the study. Cytochrome c-reductase did not show any significant

change during the whole period of study in all groups. From the present study, we

can conclude that the co-carcinogenic effect of S. mansoni infection might not be

through modulating renal cytochrome P-450 or cytochrome c-reductase in particular





16

but may be through other mechanisms and/or that the repair enzyme system itself is

the target of the ultimate carcinogen or schistosomal toxins.

Bull Alex Faculty Med: XXXVII (3), 379-84 (2001)

Radioimmunoasssay of plasma glutathione S-transferase-pi as a tumour marker in hepatocellular carcinoma.

Morsi MI, Abdel-Moneim NA, Hassan AA, Al-Sharaky AS

ABSTRACT The main objective of this work was the radioiodination of GST-Pi and to use it as

the tracer of our laboratory-manufactured RIA kit to measure the plasma GST-Pi

levels in all studied groups. Serum AFP was also measured using a ready to use

IRMA kit. The diagnostic and prognostic values of both plasma GST-Pi and serum

AFP were evaluated and compared in HCC patients. This study included 3 groups:

the control group, the benign group (SHF) and the malignant group (HCC).

GST-Pi was radioiodinated efficiently; the labeling yield and the specific activity of

our 125I-labeled GST-Pi were 89.2% and 89.3 µCi/ µg, respectively. Before

chemotherapy, the levels of plasma GST-Pi and serum AFP were significantly higher

in the HCC group than those in the control and SHF groups. After 3 cycles of

chemotherapy, the plasma levels of GST-Pi were significantly decreased; but the

serum levels of AFP were non-significantly increased. The ROC curve for plasma

GST-Pi was above ROC curve for serum AFP. The pathological differentiation was

significantly and directly correlated with the plasma level of GST-Pi, but non-

significantly correlated with the serum level of AFP. The tumor size showed no

correlation with the plasma level of GST-Pi, but non-significantly correlated with the

serum levels of AFP.

Both of plasma GST-Pi and serum AFP can be used as a diagnostic tumor markers

in HCC patients; but plasma GST-Pi is superior to serum AFP in HCC diagnosis.

Plasma GST-Pi can be also used as a prognostic tumor marker in HCC patients.





17

Int. J. Cancer: 82, 789–794 (1999)

Biomonitoring of N-nitroso Compounds, Nitrite and Nitrate in the Urine of Egyptian Bladder Cancer Patients with or without Schistosoma hematobium

Infection.

, Shehata M. El Sewedy, Tousson assanAshraf A.M. H, Mohamed A. Abdel Mohsen

Aboul-Azm, Cinzia Magangnotti, Roberto Fanelli, Luisa Airoldi

ABSTRACT The excretion of nitrate, nitrite, apparent total N-nitroso compounds and volatile

nitrosamines was measured in 24 hr urine from 61 Egyptians, divided into 4 groups:

controls, Schistosoma haematobium-infected patients and bladder cancer patients

with and without a history of schistosomal infection. Urinary nitrate in S.

haematobium-infected patients was significantly higher than in the other 3 groups.

Nitrite was below the detection limit of the method (≤ 015 mg/mg creatinine) in all but

one of the control samples. S. haematobium infection significantly increased urinary

nitrite to 0.9 ± 1.16 mg/mg creatinine (mean ± SD, p = 0.001). In both bladder cancer

groups, nitrite was about 20 times that in S. haematobium-infected patients without

bladder cancer. Excretion of apparent total N-nitroso compounds paralleled that of

nitrite. Overall, a good correlation was observed between these 2 variables (r = 0.71,

p = 0.0001). N-nitrosodimethylamine was present in all the samples analyzed. S.

haematobium

infection significantly increased urinary N-nitrosodimethylamine level compared with

that of controls (4.02 ± 1.61 and 2.04 ± 2.97 ng/mg creatinine, respectively, p =

0.01). Among

cancer patients, N-nitrosodimethylamine was higher than in controls only in those

with schistosomal infection. The presence of N-nitroso compounds and N-

nitrosodimethylamine

in the urine of S. haematobium-infected patients both before and after the

development of cancer, and the observation that these compounds also occur in

bladder cancer patients

with no history of schistosomal infection, suggest that these compounds might have

a role not only in the initiation of the carcinogenic process, but also in its progression.





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Cancer Letter: 121(1), 19-23 (1997)

Glutathione S-transferase activity and glutathione content in human bladder carcinoma associated with schistosomiasis: comparison with uninvolved

surrounding tissues.

.Airoldi L, Sewedy SM-el, D'Incalci M, Codegoni AM, Tagliabue G, Hassan AA

ABSTRACT Glutathione (GSH), glutathione S-transferase (GST) activity and GSTpi expression were measured in 10 human bladder tumors and adjacent uninvolved specimens from Egyptian patients with a history of schistosomal infection. GSH was higher in the tumor than in surrounding uninvolved tissue (not significant). Total GST activity per mg tissue protein and GSTpi expression were higher in tumor tissues (P < 0.05) than in uninvolved tissues. There was a positive correlation between GST activity and GSH content and between total GST activity and GSTpi expression in both

tumor and uninvolved tissues.

Toxicol Environ Chem: 55, 73-81 (1996)

Pollution of ground and drinking water with volatile organic compounds: solid-phase microextraction and GC/MS analysis.

, Benfenati E, Facchini G, Fanelli Rn AAMHassa

ABSTRACT Solid-phase microextraction ( SPME) is a recent method for extracting organic compounds from aqueous samples and does not require solvents. The extracting phase of a coating of a polydimethylsiloxane (PDMS) fiber assembles on a needle

within a syringe. We used SPME to set up an analytical method for some volatile organic compounds (VOCs) in water. The gas chromatographic-mass spectrometric (GC-MS) technique, in the SIR mode (Selected Ion recording), was used for detecting and quantification. We utilized standard mixture EPA VOC Mix 7 to prepare the dilutions for calibration curves and to test linearity, sensitivity and stability of the method, obtaining good results. The sensitivity achieved by this method is lower than 2 ppb (ca. 1.25 µg/l) for all the tested compounds. We also applied this technique to drinking water real

samples.

Physiology & Behavior: 57(3), 515-22 (1995)





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Metabolic inter-organ relations by exercise of fed rat: carbohydrates, ketone body, and nitrogen compounds in splanchnic vessels.

.Kido R, Hassan AM, Minatogawa Y, Hirai T

ABSTRACT Fed rats were exercised until exhaustion by almost 65% VO2max on a treadmill. In 2.5 min after the exercise, blood was collected from various vessels of the splanchnic bed. Metabolites, glucose, lactate, ketone body, and nitrogen compounds in the plasma, were measured. Glucose excretion from the liver was increased by exercise, but was not significant. The absorption by the kidney decreased to 30% by exercise. Lactate was highly absorbed by the kidney, lower limbs, and digestive tract by exercise. Exercise caused a 200-300% increase of the plasma beta-hydroxybutyrate, but the absorption by the kidney and the lower limbs was decreased. These data suggest that glucose is a good carbon source for the recovery, and that lactate is more useful than glucose, but ketone body is less effective at a very early recovery phase under fed condition. Amino acid balances in each organ except digestive tract were positive showing anabolic conditions of these organs even after exhaustive exercise at fed condition. Most amino acid concentrations in the plasma tended to decrease to 60-90% by exercise. Amino acids were excreted from the digestive tract, and were eventually absorbed by the liver in both rested and exercised rat. The digestive tract, therefore, seems to be a primary amino acids pool to supply them to the liver during the inter meal. Urea excretion from the liver was more than the absorbed ammonia showing that active deamination from amino acids was carrying on. The resulted carbon skeletons of the

amino acids might be used for the gluconeogenesis in the liver.

J Egypt Public Health Assoc: 70(3-4), 431-47 (1995)

Changes in serum lipid profile and esterases of rats after sublethal daily doses of dimethoate. Hassan AA, El-Khalili MM, Hussein NG, Kido R.

ABSTRACT Male Wistar rats were injected by dimethoate (10 mg/0.5 ml) daily for 8 successive days. Controls received the same amount of saline., A group of 5 rats were anesthetized at 0, 2, 4, 6 and 8 days of injection. Blood was withdrawn from heart. serum lipid components and 4 species of serum esterases were assayed for each group. A general decrease in the activities of serum esterases was observed. A marked decrease also was observed in lipids profile during the 8 days course of experiment.

Arch Environ Contam Toxicol: 27(2), 256-9 (1994)

Changes of some serum parameters and amino acids content in rats after chronic sublethal doses of dimethoate.

.Kido R, Hirai T, Minatogawa Y, Hassan AA

ABSTRACT





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An intraperitoneal repeated dose of dimethoate (O,O-dimethyl S-[N-methylcarbamoyl methyl] phosphorodithioate) was injected to male Wistar rats for 8 successive days. Body weight and liver weight decreased significantly while liver/body weight ratio and mg protein/g liver remained unchanged. Twenty serum amino acids were assayed in blood samples collected from hepatic vein, portal vein, and aorta. Most of the measured amino acids showed a tendency of decrease compared to control. The decrease was significant only for taurine, serine, threonine, methionine, tyrosine, histidine, and arginine but not for the rest of the amino acids measured. Glucose, 3-hydroxybutyrate, urea, ammonia, and lactate were measured in the same serum samples. These serum parameters showed a tendency of decrease (except urea) of

them from control but the decrease was not significant.

J Environ Sci Health: 26(3), 333-8 (1991)

In vitro effect of dimethoate on the activity of tryptophan pyrrolase in rat liver.

.Kido R, Minatogawa Y, Sewedy SM-lE, Hassan AA

ABSTRACT Total and holo-enzyme activities of tryptophan 2,3-dioxygenase were measured in vitro in the presence and absence of the organophosphorous insecticide, dimethoate. Addition of dimethoate to the reaction mixture decreased the activities of both total and holo-forms. Total and holo-enzyme activities were decreased by 34% and 26%, respectively, by 1 mM dimethoate. On the other hand, 5 mM dimethoate resulted in 56% and 34% inhibition to total and holo-enzyme activities, respectively. Lineweaver-Burk plot of the total-enzyme activity at different tryptophan concentration in the presence of 2 mM dimethoate gave uncompetitive type of

inhibition.


Effects of some insecticides on several enzymes of tryptophan metabolism in rats.

.Kido R, Minatogawa Y, Sewedy SM-lE, Hassan AA

ABSTRACT The activities of tryptophan 2,3-dioxygenase, indoleamine 2,3-dioxygenase, kynurenine 3-hydroxylase, kynureninase, kynurenine transaminases, and pyridoxal phosphokinase in the liver, kidney and lung rats were measured after administration of a single dose and repeated doses of dimethoate, carbaryl and fenvalerate, respectively. Ten percent LD50 of each insecticide was orally administered to a rat for a single dose, while 5% LD50 was orally given for five consecutive days as repeated doses. The control animals received the same volume of vehicle (polyethylene glycol 300). Body weight and organs weight losses were recognized only after repeated doses of dimethoate, while protein content remained constant compared to control animals. Repeated administration of dimethoate caused significant decrease in the





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activity of kynurenine 3-hydroxylase, kynurenine-pyruvate transaminase, kynurenine- pyruvate transaminase (24.5% in kidney) and pyridoxal phosphokinase (36.1% in liver). Repeated doses of carbaryl resulted in a significant decrease in the activity of apo-tryptophan 2,3-dioxygenase (42.8%), kynurenine-2-oxoglutarate transaminase (40% in liver), kynurenine-pyruvate transaminase (30.6% in liver), and serine-glyoxylate transaminase (47.9% in liver). Externally added insecticides at different concentrations to the incubation mixture resulted in an inhibition to tryptophan 2,3-dioxygenase, while the other enzymes examined showed no change in their

activities.


In vivo studies on the effect of some insecticides on the hepatic activities of L-tryptophan 2,3-dioxygenase and pyridoxal phosphokinase of male mice.

El-Toukhy MA, Ebied SA, Hassan AA, El-Sewedy SM.

ABSTRACT The effect of Dimethoate, Carbaryl and Permethrin on the activities of liver L-tryptophan 2,3-dioxygenase (total-, holo-, and apo-forms) and pyridoxal phosphokinase of male mice was investigated. Dimethoate inhibited both enzyme systems after single and repeated dose treatment; except the dioxygenase holo-enzyme after repeated doses. Single dose of Carbaryl treatment inhibited the pyridoxal phosphokinase, total and holo-enzyme of L-tryptophan dioxygenase. However, the repeated dose treatment do not affect both enzyme systems. Permethrin inhibited only total-, holo- and apo-enzymes of L-tryptophan dioxygenase whereas repeated administration has no significant effect on both enzymes. The data indicate that these insecticides may induce abnormal tryptophan metabolism

through which some endogenous bladder carcinogens are formed.




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C.V Ashraf Hassan 2010...Clinical Biochemistry course ( Enzymes, metabolism and metabolic disorders ) for undergraduate students at Faculty of Health Sciences, Jazan University, Kingdom