[Show abstract][Hide abstract] ABSTRACT: Current estimates indicate that the hepatitis C (HCV) is the leading cause of mortality around the world, with infection rates steadily increasing in Egypt. The dual therapy for this silent epidemic with pegylated-interferon-α2b/ribavirin has markedly improved the success rates in genotype-4 patients. It was reported that apoptosis plays a vital mechanistic role in limiting viral replication. P53, a key regulator of apoptosis, induces CD95 gene expression and subsequently initiates apoptotic cascade to be activated. The current study examined the impact of P53 rs1042522 and CD95 rs1800682 polymorphisms on the treatment response. Three groups of 240 volunteers were enrolled in this study; 86 in sustained virological responders group, 74 in non-responders group, and 80 in controls group. All patients had HCV genotype-4a and were interferon treatment naïve. Quantizations of HCV-RNA by qRT-PCR and histological scores were performed for all patients. In addition, genotyping of HCV-RNA, P53 rs1042522 Arg/Pro and CD95 rs1800682 A/G polymorphisms were investigated in all subjects. It was resulted that P53 Pro/Pro homozygous genotype has high significant increase, while CD95 A/A homozygous genotype has high significant decrease when comparing non-responders with responders. Finally, it was concluded that Pro variant of P53 rs1042522 may be used as a genetic predictor for non-responsiveness, while A/A variant of CD95 rs1800682 may be used as a sensitive biomarker for responsiveness to antiviral therapy of HCV genotype-4a infection. In addition, low prolactin, high total testosterone, and high growth hormone levels may provide promising biomarkers for early prediction of the response when associated with these genetic polymorphisms.
[Show abstract][Hide abstract] ABSTRACT: Hepatitis C virus (HCV) infection is associated with the development of hepatocellular carcinoma (HCC). The molecular mechanisms of HCV-associated carcinogenesis are unknown. We aim to investigate the alteration of the total nuclear DNA content (ploidy) in different histopathological liver tissues infected with HCV and their relation to the seropositivity of HCV RNA. Methods: Blood and liver tissues were collected from 26 patients. Diagnosis was carried out according to clinical and pathological examinations by specialized physicians. HCV RNA was detected in patients' sera and tissue samples by RT-PCR. To examine nuclear DNA ploidy, liver tissues were stained with blue Fulgen using the image analysis techniques. Finally, the patients' DNA content was examined by histochemical analysis depending on the optical density of DNA from liver biopsies using the grey image menu in each specimen. Results: The HCV RT-PCR results demonstrated that 13/26 (50%) patients had detectable HCV RNA in their sera samples while 18/26 (69%) had detectable HCV RNA in liver tissues. The DNA content from those patients measured by image cytometry showed a high level of alteration of nuclear DNA ploidy and proliferation in liver tissues with HCC, less alteration of nuclear DNA ploidy in cirrhotic patients, and least proliferation nearly normal in liver fibrosis patients. Moreover, the results of histochemical analysis confirmed the DNA image cytometry results and showed that positive HCV RNA liver tissues had more DNA ploidy than negative HCV RNA liver tissues with statistical significance (p-value < 0.05). Conclusions: HCV positive liver tissue had alterations in DNA content (ploidy) which may lead to liver disease progression, malignant transformation of the liver cells and development of hepatocellular carcinoma.
Full-text · Article · Jan 2015 · Clinical laboratory
[Show abstract][Hide abstract] ABSTRACT: A Series of chalcone derivatives containing pyrazole ring was prepared and their cytotoxicity against different human cell lines, including breast (MCF-7), colon (HCT-116) liver (HEPG2) cell lines, as well as normal melanocyte HFB4 was evaluated. Two of these chalcone derivatives with different IC50 and chemical configuration were chosen for molecular studies in detail with MCF-7 cells. Our data indicated that the two compounds prohibit proliferation, angiogenesis, cell cycle progression and induce apoptosis of breast cancer cells. This inhibition is mediated by up regulation of tumor suppressor p53 associated with arrest in S-G2/M of cell cycle. This work provides a confirmation of antitumor activity of the novel chalcones and assists the development of new agents for cancer treatment.
Full-text · Article · Nov 2014 · Anti-Cancer Agents in Medicinal Chemistry (Formerly Current Medicinal Chemistry - Anti-Cancer Agents)
[Show abstract][Hide abstract] ABSTRACT: Pseudomonas aeruginosa TMN was used to produce rhamnolipid (RL) from a variety of carbon and nitrogen substrates. The most favorable carbon sources for RL production were glucose and glycerol (both at 40 g/L), giving a RL yield of 0.3 and 0.25 g/L, respectively. Meanwhile, sodium nitrate appeared to be the preferable nitrogen source, resulting in a RL production of 0.34g/L. Rhamnolipid production from P. aeruginosa TMN was affected by temperature, pH and agitation rate, with 37 °C, pH 7 and 200 rpm agitation favorable for rhamnolipid production. Fourier transform infrared spectroscopy (FTIR), nuclear magnetic resonance (NMR) and electro spray ionization–mass spectrometry (ESI–MS) analyses indicated that the purified product contained one type of commonly found rhamnolipid, which is L-rhamnosyl-L-rhamnosyl-β- hydroxydecanoyl-β-hydroxydecanoate. The rhamnolipid product can reduce the surface tension of water to 34 mN/m with a critical micelle concentration of nearly 18.75 mg/L and emulsified kerosene by 46%. P. aeruginosa TMN strain is a potential source of rhamnolipid biosurfactant, which could be used for the development of bioremediation processes in the marine environment.
Full-text · Article · Oct 2014 · Brazilian Journal of Chemical Engineering
[Show abstract][Hide abstract] ABSTRACT: The hepatitis C virus (HCV), the main cause of morbidity and mortality, is endemic worldwide. HCV causes cirrhosis and other complications that often lead to death. HCV is most common in underdeveloped nations, with the highest prevalence rates in Egypt. Tumor suppressor gene (P53) induces the expression of apoptotic antigen-1 gene (APO-1) by binding to its promoter for mediating apoptosis; an important mechanism for limiting viral replication. This study aims at investigating the impact of P53 72 Arg/Pro and APO-1 -670 A/G polymorphisms on HCV genotype 4a susceptibility. Two hundred and forty volunteers were enrolled in this study and divided into two major groups; 160 HCV infected patient group and 80 healthy control group. HCV patients were classified according to Metavir scoring system into two subgroups; 72 patients in F0/1-HCV subgroup (patients with no or mild fibrotic stages) and 38 patients in F3/4-HCV subgroup (patients with advanced fibrotic stages). Quantification of HCV-RNA by qRT-PCR and fibrotic scores as well as genotyping of HCV-RNA, P53 at 72 Arg/Pro, and APO-1 at -670 A/G were performed for all subjects. It was resulted that F0/1-HCV patients have significant differences of P53 at 72 (Pro/Pro and Arg/Arg) genotypes and dominant/recessive genetic models as well as APO-1 -670 A/A genotype and dominant genetic model as compared to F3/4-HCV patients. Moreover, HCV patients have significant differences of P53 at 72 (Pro/Pro) genotype and recessive genetic model as well as APO-1 -670 A/A genotype and dominant genetic model as compared to those of healthy individuals. Finally, it was concluded that P53 rs 1042522 (Pro/Pro and Arg/Arg) genotypes and APO-1 rs 1800682 A/A genotype may be potentially used as sensitive genetic markers for HCV genotype 4a susceptibility.
[Show abstract][Hide abstract] ABSTRACT: Objectives: the current study is concerned with investigating the circulating prolactin and testosterone in HCV genotype-4a patients.
Moreover, the present study aimed at exploring the impact of the pegylated-interferon-a plus ribavirin (PEG-IFN-a/RBV) dual therapy
on the levels of these hormones.
Design and Methods: Eighty consecutive outpatients with chronic HCV genotype-4a and 40 healthy volunteers were enrolled in
the prospective study. Consequently, HCV genotype-4a patients have been divided, depending on the responsiveness ability to the
antiviral therapy, into 43 sustained virological responders (SVR) and 37 non-responders (NR). RNA was isolated from sera for HCV
genotyping and viral load measuring, to confirms the seropositively results obtained from standard viral antibody tests, by real time
polymerase chain reaction (qRT-PCR). Prolactin and total testosterone concentrations were quantitatively determined using the
commercially available high-sensitivity immunoassays. Hepatic profile measurements were performed in HCV genotype-4 patients
treated with PEG-IFN-a/RBV antiviral therapy and healthy controls.
Results: There is a significant increase of prolactin concentrations when comparing chronic HCV genotype-4a infected patients
with healthy subjects; regardless of gender. Besides, prolactin concentrations decreased significantly after antiviral therapy in
SVR as compared to NR; regardless of gender. While, there are high significant decreases of post-treatment total testosterone
concentrations when comparing males NR with SVR.
Conclusions: We conclude that HCV genotype-4a patients have high prolactin concentrations; regardless of gender. Besides, there
is an early normalization of an abnormal prolactin level as an indicative response to antiviral treatment. While, males NR patients
have lower post-treatment total testosterone levels when comparing with SVR.
Full-text · Article · May 2014 · The Open Biochemistry Journal
[Show abstract][Hide abstract] ABSTRACT: Viral infection with hepatitis C virus (HCV) has a high propensity in becoming chronic and it is the major cause of hepatocellular carcinoma (HCC) worldwide. This review was basically established to illustrate the putative role of the P53 gene Arg72Pro polymorphism on various cancer models and viral infections, focusing on HCV and HCC incidences. Authors studied the 72 G/C single base substitution of P53 gene at codon 72 using various polymorphic techniques. Intriguingly, authors investigated that the P53 codon 72 plays a crucial role as risk factor in several cancer models. Others found that there is no association between codon 72 genotypes and HCV disease severity or liver cancer. Moreover, the lack of a significant relationship between this polymorphism and risk of HCC shows that it does not predispose towards hepatocarcinogenesis and the frequent loss of the proline allele in HCV-associated carcinogenesis of the liver plays some critical role in hepatocarcinogenesis. Amazingly, there is a significant correlation between male homozygotes for P53 72Pro with HCV type 1b infection. However, there was no significant difference between the P53 polymorphism and HCV genotypes 2a and 2b. It was concluded that the P53 gene polymorphism at codon 72 has been investigated as potential risk factor in several cancer models and HCV infections.
Full-text · Article · Jan 2014 · Indian Journal of Clinical Biochemistry
[Show abstract][Hide abstract] ABSTRACT: The prevalence of hepatitis C virus (HCV) infection varies across the world, with the highest percent of infections reported in Middle East, increasingly in Egypt. The current study aimed at examining the bio-statistical correlation and multiple regression analyses of pituitary growth hormone (GH) and liver activities among HCV genotype-4 patients treated with PEG-IFN-α plus RBV therapy. Herein, the current study was conducted on 100 HCV genotype-4 infected patients and 50 healthy controls. Patients received PEG-IFN-α/RBV for 24 weeks. Host RNA was isolated from patients’ sera for HCV genotyping and viral load determination. Moreover, the enzymatic activities of the liver, AFP, GH, PT, and CBC were performed in all volunteers. The present study resulted that the activities of the hepatic enzymes among HCV genotype-4 patients correlated together significantly. While, human GH showed a significant positive regression with pre-treatment ALT concentration in responders. Furthermore, multiple regression analysis for GH showed a significant positive correlation with pre-treatment ALT in HCV genotype-4 infected patients. We concluded that there were a putative significant relation between GH and pre-treatment ALT activity in HCV infection and response to IFN-based therapy.
No preview · Article · Oct 2013 · Indian Journal of Clinical Biochemistry
[Show abstract][Hide abstract] ABSTRACT: The main objective of the current study is to examine the role of the statistical relation between BCL2 gene (Ala43Thr) single nucleotide polymorphism and growth hormone (GH1) levels in Egyptian HCV genotype-4 patients before and after treatment with pegylated interferon plus ribavirin. Eighty patients with HCV genotype-4 and 40 healthy volunteers as controls were enrolled in the prospective study. Gene polymorphism of BCL2 (Ala43Thr) using PCR-RFLP technique and GH1 concentrations using ELISA procedure were measured for all patients and controls. The present study resulted that Responder HCV genotype-4 Patients, with BCL2 43Ala genotype, have high significant increase in pre-treatment GH1 levels (>1 ng/ml); which represent normal levels, as compared to non-responders pre-treatment GH1 levels (<1 ng/ml); which represent low concentrations. We concluded that HCV genotype-4 patients who have normal GH1 concentrations and BCL-2 43Ala genotype can successfully achieve response to interferon based therapy.
No preview · Article · Oct 2012 · Indian Journal of Clinical Biochemistry
[Show abstract][Hide abstract] ABSTRACT: A newly synthesized series of chalcone derivatives containing pyrazole rings were synthesized and evaluated for their cytotoxic activities in vitro against several human cancer cell lines. Most of the prepared compounds showed potential cytotoxicity against human breast cancer cell lines MCF-7, HEPG-2, and HCT-116. Also the compounds were evaluated as antimicrobial agents. The three compounds 3, 4, and 5 were proved to be better anticancer agents than the positive standard doxorubicin with IC50 values (4.7, 4.4, and 3.9 μg/ml) against the same human cancer cell lines, whereas compounds 5 and 6 showed the most active antimicrobial compounds in comparison to the other chalcones.
[Show abstract][Hide abstract] ABSTRACT: A novel potyvirus we call Gazar virus Y (GVY) was isolated from diseased carrot plants in Egypt. It produced symptoms similar to those caused by carrot virus Y. Its particles were visualized in leaf extracts using negative stain, and leaf tissues from carrot plants infected with GVY had many ultra-structural differences from healthy ones. Chemical analyses of healthy and infected carrot roots showed that carrot roots infected with GVY had increased dry weight and decreased moisture, total ash, total carbohydrates, total fibres, vitamin C, β-carotene, vitamin A and the minerals of K, Mg, Na, P and Ca compared to the healthy carrot roots. Molecular methods, including reverse transcription-polymerase chain reaction (RT-PCR), were used to amplify a 335 bp cDNA fragment transcribed from infected plant extracts using degenerate oligonucleotide primers specific for potyviruses. The amplified cDNA was cloned into pGEM®-T Easy vector, and transformed into Escherichia coli (E. coli) strain DH5α . The recombinant inserts were sequenced. The nucleotide sequence (GenBank Accession Code GQ148776) was matched against viral nucleotide sequences in the GenBank database. It is most closely related to viruses of the celery mosaic virus group of potyviruses, but is distinct from all and represents a sister lineage to all others described so far.
[Show abstract][Hide abstract] ABSTRACT: With the purpose of evaluating the ability of Pd(II) complex to interact with DNA molecule as the main biological target, two new complexes [Pd(bpy)(OH(2))(2)] (1) and [Pd(Phenpip)(OH(2))(2)] (2), where (bpy=2,2'-bipyridine; Phenpip=1-phenylpiperazine), have been synthesized and the binding properties of these complexes with CT-DNA were investigated. The intrinsic binding constants (K(b)) calculated from UV-Vis absorption studies were 3.78×10(3)M(-1) and 4.14×10(3)M(-1) for complexes 1 and 2 respectively. Thermal denaturation has been systematically studied by spectrophotometric method and the calculated ΔT(m) was nearly 5°C for each complex. All the results suggest an electrostatic and/or groove binding mode for the interaction between the complexes and CT-DNA. The redox behavior of the two complexes in the absence and in the presence of calf thymus DNA has been investigated by cyclic voltammetry. The cyclic voltammogram exhibits one quasi-reversible redox wave. The change in E(1/2), ΔE(p) and I(pc)/I(pa) supports that the two complexes exhibit strong binding to calf thymus DNA. Further insight into the binding of complexes with CT-DNA has been made by gel electrophoresis, where the binding of complexes is confirmed through decreasing the intensity of DNA bands. The two complexes have been screened for their antimicrobial activities using the disc diffusion method against some selected Gram-positive and Gram-negative bacteria. The activity data showed that both complexes were more active against Gram-negative than Gram-positive bacteria. It may be concluded that the antimicrobial activity of the compounds is related to cell wall structure of bacteria.
No preview · Article · Jul 2012 · Spectrochimica Acta Part A Molecular and Biomolecular Spectroscopy
[Show abstract][Hide abstract] ABSTRACT: In this study two new complexes [Cu(bpy)(Gly)Cl]·2H(2)O (1) and [Cu(dpa)(Gly)Cl]·2H(2)O (2) (bpy=2,2'-bipyridine; dpa=2,2'-dipyridylamine, Gly=glycine) have been synthesized and characterized by elemental analysis, IR, TGA, UV-vis and magnetic susceptibility measurements. The binding properties of the complexes with CT-DNA were investigated by electronic absorption spectra. The intrinsic binding constants (K(b)) calculated from UV-vis absorption studies were 1.84 × 10(3) M(-1) and 3.1 × 10(3) M(-1) for complexes 1 and 2 respectively. Thermal denaturation has been systematically studied by spectrophotometric method and the calculated ΔT(m) was nearly 5 °C for each complex. All the results suggest that the interaction modes between the complexes and CT-DNA were electrostatic and/or groove binding. The redox behavior of the two complexes was investigated by cyclic voltammetry. Both complexes, in presence and absence of CT-DNA show a quasi-reversible wave corresponding to Cu(II)/Cu(I) redox couple. The change in E(1/2), ΔE and I(pc)/I(pa) ascertain the interaction of complexes 1 and 2 with CT-DNA. Further insight into the binding of complexes with CT-DNA has been made by gel electrophoresis, where the binding of complexes is confirmed through decreasing the mobility and intensity of DNA bands. In addition, the antitumor activity of the complexes was tested on two cancer cell lines; the breast cancer (MCF7) and the human hepatocellular carcinoma (HEPG2), as well as one normal cell line; the human normal melanocytes (HFB4). The results showed that complex 1 was more potent antitumor agent than complex 2. The in-vitro antimicrobial activity of the two complexes was carried out using the disc diffusion method against different species of pathogenic bacteria and fungi. The activity data showed that complex 2 was more active in inhibiting the growth of the tested organisms.
No preview · Article · Nov 2011 · Spectrochimica Acta Part A Molecular and Biomolecular Spectroscopy
[Show abstract][Hide abstract] ABSTRACT: Thaumatin-like proteins (TLPs) constitutes a homogeneous family, members of which are produced by plants in response to different kinds of stress. NP24 protein is one of such salt-induced protein from tomato (Solanum lycopersicum) and it belongs to TLPs family. NP24 is a 24 kDa (207 amino acid) antimicrobial TLP found in tomato fruits. One isoform (NP24I) of NP24 was discovered in the outer pericarp of tomato fruit and is reported to play a possible role in ripening of the fruit in addition to its antimicrobial activity. In this study, the total RNA was isolated successfully from the outer pericarp of ripe (red) tomato fruit. cDNA was prepared and the gene coding for NP24I protein was amplified using conventional polymerase chain reaction (PCR). The gene was then cloned into Mach1 (TM)- T1 (R) Escherichia coli cells, then subcloned into the over-expression vector pET-28a (+) using BL21 expression bacteria. Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) analysis showed that the gene was over-expressed in E. coli as inclusion bodies. Optimization of the recombinant NP24I protein solubility was achieved by cold induction through decreasing both expression temperature and Isopropyl-beta-thio galactopyranoside (IPTG) concentration. The recombinant NP24I protein was purified using Ni-NTA resin, and then the antimicrobial activity of the purified recombinant NP24I protein was tested. The aims of this work were to study the cloning and expression of NP24 protein from local tomato cultivar in a prokaryotic system and to test the activity of the recombinant NP24, as well as to prove the activity of native protein on the bacterial as well as fungal growth.
Full-text · Article · Oct 2011 · AFRICAN JOURNAL OF BIOTECHNOLOGY
[Show abstract][Hide abstract] ABSTRACT: Interferon used in the treatment of hepatitis C virus (HCV) patients stimulates the expression of a number of host genes encoding enzymes with antiviral activities, including myxovirus resistance gene- 1 ( Mx1 ). Mx1 gene was found to have a single nucleotide polymorphism (SNP) at position -88 in the promotor region that affect the expression of Mx 1 protein and was suggested to be associated with the response of HCV. In this study, we assessed the relation between the SNP in the Mx1 gene and the responsiveness of Egyptian HCV patients to pegylated interferon and ribavirin treatment along with other host-related and virus-related predictors of treatment outcome. We genotyped the biallelic G/T SNP in the promoter region of Mx1 gene at position -88 from the transcription start site by restriction fragment length polymorphism (RFLP) in 42 interferon treatment-naïve Egyptian patients that were treated with pegylated interferon and ribavirin. We found that Mx1 nt-88 SNP is not significantly correlated to achieving sustained virological response (SVR) after pegylated interferon alpha and ribavirin combined treatment. We conclude that Mx1 gene polymorphism at codon nt-88 cannot be considered as biological marker to potentially identify responders and non-responders of HCV patients to achieve a sustained virological response to treatment with interferon (IFN) in combination with ribavirin.
Full-text · Article · Oct 2011 · AFRICAN JOURNAL OF BIOTECHNOLOGY
[Show abstract][Hide abstract] ABSTRACT: To explore the impact of response to interferon and ribavirin antiviral therapy on human growth hormone (hGH) levels in Egyptian chronic hepatitis C genotype-4 infected patients.
We studied eighty Egyptian HCV infected patients visiting outpatient clinics of Tropical Medicine and Hepatology Department, El-Kasr El-Aini Hospital, Cairo University, Egypt. HCV patients received treatment of interferon and ribavirin combination therapy for 24 weeks. Clinical, virological, histological characteristics, and biochemical tests including; liver function tests (ALT and AST), prothrombin time (PT), alpha fetoprotein (AFP), complete blood picture (CBC), and hGH were monitored in hepatitis C genotype-4 infected patients before and after interferon therapy, and healthy controls.
Chronic HCV genotype-4 infected patients have high significant decrease of hGH as compared to healthy control individuals. In addition to, there was high significant increase of hGH in responders as compared to non-responders after treatment.
We concluded that Egyptian HCV genotype-4 infected patients have growth hormone insufficiency. Besides, we found that response to interferon/ribavirin treatment has an impact on growth hormone levels.
No preview · Article · Sep 2011 · Clinical biochemistry
[Show abstract][Hide abstract] ABSTRACT: Interferon used in the treatment of hepatitis C virus (HCV) patients stimulates the expression of a number of host genes encoding enzymes with antiviral activities, including myxovirus resistance gene-1 (Mx1). Mx1 gene was found to have a single nucleotide polymorphism (SNP) at position -88 in the promotor region that affect the expression of Mx 1 protein and was suggested to be associated with the response of HCV. In this study, we assessed the relation between the SNP in the Mx1 gene and the responsiveness of Egyptian HCV patients to pegylated interferon and ribavirin treatment along with other host-related and virus-related predictors of treatment outcome. We genotyped the biallelic G/T SNP in the promoter region of Mx1 gene at position -88 from the transcription start site by restriction fragment length polymorphism (RFLP) in 42 interferon treatment-naïve Egyptian patients that were treated with pegylated interferon and ribavirin. We found that Mx1 nt-88 SNP is not significantly correlated to achieving sustained virological response (SVR) after pegylated interferon alpha and ribavirin combined treatment. We conclude that Mx1 gene polymorphism at codon nt-88 cannot be considered as biological marker to potentially identify responders and non-responders of HCV patients to achieve a sustained virological response to treatment with interferon (IFN) in combination with ribavirin. Key words: Hepatitis C virus (HCV), interferon (IFN), myxovirus resistance protein (Mx1 protein), myxovirus resistance gene-1 (Mx1 gene), single nucleotide polymorphism (SNP).
[Show abstract][Hide abstract] ABSTRACT: The prevalence of hepatitis C virus (HCV) infection varies across the world, with the highest number of infections reported in Egypt. BCL-2 gene polymorphism at codon 43 (127G/A) has been found to be a reliable and sensitive marker for the prediction of response to interferon therapy during viral infections. This study examined the correlation of BCL-2 gene polymorphism with the response to treatment with pegylated-IFN-alfa2b and ribavirin. Eighty patients with type 4 HCV and 40 healthy volunteers as controls were enrolled in a prospective study. Quantification of HCV-RNA by real-time PCR was performed for every patient, and gene polymorphism of BCL-2 (ala 43 Thr) was performed for all patients and controls. There was a statistically significant difference between non-responder patients and control group as regards the 43Thr genotype and allele (P<0.05). Also, there was a statistically significant difference between responders and non-responders (P<0.05) as regards 43Thr genotype and alleles. We conclude that BCL-2 gene polymorphism at codon 43 (127G/A) is a new biological marker to potentially identify responders and non-responders of HCV genotype 4 patients to achieving a sustained virological response to treatment with IFN in combination with ribavirin.
Full-text · Article · Mar 2011 · Clinica chimica acta; international journal of clinical chemistry