Vol. 39, July, 2012

1/39 ANTI-INFLAMMATORY EFFECT OF APIGENIN-7-NEOHESPERIDOSIDE (RHOIFOLIN) IN CARRAGEENIN-INDUCED RAT OEDEMA MODEL

Omayma A. Eldahshan and Samar S. Azab*

Pharmacognosy Department, Faculty of Pharmacy, Ain Shams University,

Cairo, Egypt

*Pharmacology and Toxicology Department, Faculty of Pharmacy, Ain shams University, Cairo, Egypt

Flavonoids are normal constituents of the human diet and are known for a variety of biological activities. They have been reported to bring benefits in lowering inflammation and oxidative stress. The present investigation was performed first, to evaluate the anti-inflammatory activity of rhoifolin and second, to search for the possible contributing mechanisms for this hypothesized effect.In the current study, the potential anti-inflammatory activity of rhoifolin was evaluated in carrageenin-induced rat oedema model. Anti-inflammatory activity was further assessed by examining prostaglandin E2 level, TNF-α release and the total antioxidant capacity. Results showed that rhoifolin caused a time and reverse dose dependent reduction of carrageenin-induced rat paw oedema. Following 4 hr of treatment, rhoifolin at doses 2.5, 25 & 250 mg/kg caused a significant inhibition of rat paw edema volume by 14, 25 & 45 % respectively in comparison to the control group (74%). In addition to significantly abrogating prostaglandin E2 level, increasing doses of rhoifolin significantly diminished the TNF-α release in the inflammatory exudates. In the same animal model, rhoifolin increased the total antioxidant capacity in a reverse dose order, with the highest capacity obtained with the lowest dose tested. This study demonstrates for the first time the effectiveness of rhoifolin in combating inflammation in carrageenin-induced rat oedema model.

2/39 CHEMICAL COMPOSITION, ANTIOXIDANT AND ANTIMICROBIAL ACTIVITIES OF LEAVES ESSENTIAL OILS OF TWO MANDARIN CULTIVARS

Seham S. El-hawary, Kamilia F. Taha*, Farid N. Kirillos, Azza R. Abdel-Monem, Amira A. Mohamed*.

Department of Pharmacognosy, Faculty of Pharmacy, Cairo University,

Cairo 11562, Egypt.

*Applied Research Center of Medicinal Plants, National Organization of Drug Control and Research, Cairo, Egypt.

Leaves essential oils of two mandarin cultivars, Citrus deliciosa cv Avana apriena and Citrus reticulata cv Seedless mandarin, were prepared by hydrodistillatin. The essential oils content was 0.35% and 0.5% (v/w on fresh weight basis) in Avana apriena mandarin and Seedless mandarin, respectively. Their chemical composition was investigated by GC/MS,methyl-N-methylanthranilate being the major constituent (59.72% and 54.95 %). Monoterpene hydrocarbons constituted 37.26% and 35.33%, and limonene content was 11.83% and 7.73% in Avana apriena mandarin and Seedless mandarin, respectively. The essential oils of the studied mandarin species exhibited variable antioxidant and antimicrobial activities.

3/39 SERUM NEUTROPHIL GELATINASE-ASSOCIATED LIPOCALIN AS A MARKER OF RENAL FUNCTION IN EGYPTIAN PATIENTS   WITH CHRONIC KIDNEY DISEASE.

Omayma H.M.Sarhan and Somayh Suliman Eissa*

Departments of Biochemistry and *Internal Medicine, Faculty of Medicine for Girls, Al Azhar University.

The serum levels of Neutrophil gelatinase-associated lipocalin (NGAL) were measured in 60 adult patients with chronic kidney disease (CKD). The patients were categorized into four groups [stages 2-5] according to severity of renal impairment, and 20 apparently healthy subjects serving as a control group. Assay was carried out using an enzyme-linked immunosorbent technique. Serum NGAL showed a highly significant increase in CKD patients as compared to healthy controls. Also, a significant positive correlation was found between serum NGAL and serum creatinine, blood urea nitrogen (BUN) and urinary albumin. Meanwhile, an inverse correlation was recorded between serum NGAL and the estimated glomerular filtration rate (eGFR) in CKD patients collectively. The relation between the levels of NGAL and the severity of CKD, showed a highly significant progressive increase throughout stages 2 to 5. Furthermore, serum NGAL was inversely correlated with eGFR in early stages of CKD (stages 2&3). As kidney function worsen (stages 4&5), this correlation becomes insignificant; a finding which might reflect an implicated renal tubulo-interstitial pathology rather than glomerular dysfunction. Accordingly, determination of serum NGAL level may provide an additional accurate measure of kidney function and renal pathology in CKD.

4/39 POTENTIAL CYTOTOXIC, HEPATOPROTECTIVE AND ANTIOXIDANT ACTIVITIES OF DELONIX REGIA FLAVONOIDS

Samar S. Azab, Mohamed Abdel-Daim* and Omayma A. Eldahshan**

Pharmacology and Toxicology Department, Faculty of Pharmacy, Ain Shams University, Cairo, Egypt

*Pharmacology Department, Faculty of Veterinary Medicine, Suez Canal University, Ismailia, Egypt.

**Pharmacognosy Department, Faculty of Pharmacy, Ain Shams University,

Cairo, Egypt

The present study was designed to target four specific aims; first, Phytochemical investigation of Delonix regia extract which led to the isolation of seven flavonoid glycosides; Kaempferol 3-rhamnoside, Quercetin 3-rhamnoside, Kaempferol 3-glucoide, Kaempferol 3-rutinoside, Kaempferol 3-neohesperidoside, Quercetin 3-rutinoside and Quercetin 3-glucoside. Second, the evaluation of in-vitro cytotoxic activity of the extract, where the extract exhibited potent cytotoxic effect against HepG2 cell line. The hepatoprotective activity was measured using biochemical parameters such as serum aspartate aminotransferase, alanine amino transferase, alkaline phosphatase, total protein, albumin, total and direct bilirubin. The effective antioxidant activity of the extract was measured using superoxide dismutase, catalase, reduced glutathione and malondialdehyde levels and total antioxidant capacity. The results suggest that the extract had a dose dependent hepatoprotective and antioxidant effect. The current findings concluded that the extract of Delonix regia possessed not only a significant anticancer effect against HepG2 cells, but also an effective hepatoprotective activity.

5/39 FETUIN- A AND SOLUBLE TWEAK IN EGYPTIAN PATIENTS WITH CHRONIC KIDNEY DISEASE

Abeer I. Abd El-Fattah and Osama M. Ahmad*

Biochemistry Department, Faculty of Pharmacy (girls) and*Internal Medicine Department (Nephrology unit), Faculty of Medicine (boys) - Al Azhar University.

Defective endothelial (ED) function, an initial step in the development of atherosclerotic plaque, is prevalent in moderate to advanced chronic kidney disease (CKD). Fetuin-A is a cirulating calcium-regulatory glycoprotein that inhibits vascular calcification. Recently, it has generated much interest and may be one of the novel contributing factors for the development of ED in CKD patients. Soluble TNF–like induces of apoptosis (sTWEAK) is a recent determinant of ED and a novel predictor of mortality in dialysis patients. Little information are available about the prognostic role of fetuin A and s TWEAK as novel markers of (ED) in Egyptian patients with (CKD) so , this study was designed to; (1) Evaluate serum fetuin –A and s TWEAK as the main factors for endothelial dysfunction in Egyptian patients with different stages of CKD. (2)Study of their relation to other inflammatory markers (IL6 and hsCRP) and other metabolic and anthropometric variables. (3) Examine their association with the severity of kidney dysfunction. (4) Predict atherogenic risk by vascular ED assessment by measuring flow-mediated dilatation (FMD) and Carotid intima-media thickness (CIMT). This study included 80 individuals,divided into: I-Thirty patients of chronic kidney disease; stage 3-4 (CKD3-4 ). II-Thirty patients of chronic kidney disease; stage 5 in the end stage renal disease (ESRD), on regular haemodialysis (haemodialysis group, HD). III- Twenty age and sex matched healthy individuals served as a control group. Patients with known diabetes mellitus, congestive heart failure, myocardial infarction, malignancies and active infections were excluded. Concentrations of Fetuin-A, sTWEAK, IL-6 and hs-CRP were determined in the sera of all individuals using commercially available ELISA kits. Other biochemical factors were determined using standard methods. Vascular endothelial dysfunction assessment was done, as flow-mediated dilatation assessed by high-resolution brachial ultrasonography was performed. Carotid intima-media thickness was also estimated by ultrasonography.Our results showed that there was a highly significant decrease in fetuin-A and sTWEAK in the HD group (429 ± 22 µg/ml , 187.11± 9.9 pg/ml respectively) compared to CKD3-4 group (474 ± 20 µg/ml , 279.87± 12 pg/ml respectively) and control (608 ± 35 µg/ml , 449.32± 21 pg/ml respectively) at P = 0.0001 . In addition, Fetuin-A and sTWEAK levels were found to be significantly correlated with inflammatory biomarkers (IL6, CRP), FMD, CIMT and other metabolic and anthropometric variables.It concluded thatour results may point toward a role of both Fetuin-A and sTWEAK in CKD, as their levels were strongly associated with ED and inflammatory activation typically observed with progressive kidney failure. Additionally, both biomarkers impacted the predictability of cardiovascular outcomes; this was done dependently by measuring vascular derangements by means of (FMD and CIMT) levels. This may encourage further mechanistic research regarding the potential of Fetuin-A and sTWEAK as therapeutic targets in inflammation- or atherosclerosis-related diseases.

6/39 RELATION BETWEEN SERUM VISFATIN AND CLINICAL SEVERITY IN DIFFERENT STAGES OF RHEUMATOID ARTHRITIS

Abeer Ibrahim and *Ashraf Ismael

Biochemistry Department, Faculty of Pharmacy (girls) and *Rheumatology Department, Faculty of Medicine, Al-Azhar University.

Visfatin is a one of the recently discovered adipokine that has important pro-inflammatory and catabolic roles in rheumatoid arthritis (RA).Pro-inflammatory cytokines, such as interleukin (IL)-1, IL-6, and (TNF-α), initiate physiological changes that result in the characteristic signs of inflammation. Since inflammation is the major factor leading to structural damage, it is critical to achieve rapid suppression of inflammation to maximize disease control. Therefore, early diagnosis and treatment of RA is of paramount importance. Since, the information about the relation between visfatin and disease activity in RA patients is conflicting and little is known about its role in radiographic joint damage, the present study designed to evaluate the role of serum visfatin as a recent pro-inflammatory marker in RA according to the radiological severity scores of disease to assess the possibility of introducing serum visfatin in the diagnosis and monitoring of RA patients and correlate between its serum level and other cytokines (IL-6 and TNF-a) and other laboratory biomarkers. This study was conducted on a total number of 80 individuals, 60 of them were RA (48 females 80%, 12 males 20%) and 20 healthy subjects as a control (10 females 50%, 10 males 50%). RA patients were classified to 3 groups:Group I (severe RA): 20 RA patients, Group II (moderate RA):20 RA patients and Group III (mild RA): 20 RA patients. Diagnosis of RA was based and confirmed by the American College of Rheumatology (ACR) / (EULAR) 2010 criteria. All patients underwent clinical evaluation for disease activity assessed using a 28 joint disease activity score, (DAS-28), pain using visual analogue scale (VAS) and functional disability using the Swedish version of the Stanford health assessment questionnaire (HAQ) to calculate disability index (DI). Blood samples were obtained from patients and controls for complete blood count CBC and erythrocyte sedimentation rate ESR. The sera of patients collected for ELISA estimation of serum visfatin, IL6 and TNF-a. CRP and RF were determined by turbidimetry quantitative method. Our results showed the comparison between the RA and control groups showed that the mean serum level of visfatin, PLT, ESR, IL6, CRP and RF were significantly higher in RA patients than control group. The comparison between the mild, moderate and severe RA groups showed that the mean levels of visfatin and IL-6, VAS pain score and DI were significantly higher in severe RA group than moderate RA group which was significantly higher than mild RA group. There was a significant positive correlation between serum visfatin and {IL-6, ESR, CRP, TNFα, DAS 28 and VAS pain score} in RA group. It concluded that visfatin has a role in the pathogenesis of RA and could be considered as a disease marker in RA and a marker of radiographic joint damage and hence as a potential therapeutic target for RA. The finding of present study indicate also that serum visfatin and IL-6 might be of a valuable diagnostic value for RA, however the combined diagnosis using serum visfatin, IL-6 and RF test can improve RA diagnosis in early stage. Further studies are needed to determine the possibility of introducing visfatin as a potential therapeutic target especially in early RA to prevent erosions.

7/39 PHARMACEUTICAL CARE AND MONITORING OF DIABETIC PATIENTS IN EGYPT

Kamal A. Badr

Department of Pharmaceutics, Faculty of Pharmacy, October University for Modern Sciences and Arts (MSA), Egypt.

Screening and localizing diabetic patients in Egypt are very important goals. People that not controlled properly will be achieved through developing a pharmaceutical care program which may be easy to comprehend by health practitioners. Selected diabetic patients at several spots of Egypt in hospitals, clinics and community pharmacies were subjected to extensive monitoring of glucose level via certain questionnaires. In addition the quality control studies and tests on selected oral ant diabetic medications that are present in the Egyptian market were also investigated. Statistical data obtained by the questionnaires and survey forms filled with information obtained from diabetic patients were filtered and analyzed. Results obtained were compared with other international equivalent field work. The obtained data are quietly discriminated in the national data. Results showed that 46% of the questioned patients were males, while 54% were female patients, and 57% of the patients surveyed were diagnosed with Type 1 Diabetes, 43% are of Type 2. Unfortunately, most of patients have diabetes with other diseases especially hypertension (27%) and hypercholesterolemia (14%). Thirst, dry mouth, and shortness of breath were the most common symptoms of patients suffering; however, patient’s blood glucose level was between 300 mg/dl and 200 mg/dl. In spite of the selected patients are well educated, most of them were found to suffer of diabetic problems and uncontrolled blood glucose level. There is variability in patient’s health conditions, some of them exhibit a good health conditions and the others are fairly good and poor. Although D.M. is a common national hygienic disease the medical team has to be informed how to monitor and control the symptoms and problems with the best regulation and instructions.

8/39 NORMAL RANGE ALBUMINURIA DOES NOT EXCLUDE NEPHROPATHY IN TYPE 2 DIABETIC PATIENTS

Maha O. Mahmoud, Ola S. Mohamed*, Mahmoud M. Mahmoud** and

Ibrahim A. Emara***

Biochemistry Department, Faculty of Pharmacy, Egyptian Russian University, Cairo, Egypt

*Biochemistry Department, Faculty of Pharmacy and **Internal Medicine Department, Faculty of Medicine, Al-Azhar University, Cairo, Egypt

***Biochemistry Department,National Institute for Diabetes and Endocrinology

The aim of this work is to investigatethe role of serum and urinary neutrophil gelatinase associated lipocalin (NGAL), a tubular stress protein, in detection of early stage nephropathy in Egyptian type 2 diabetic patients. Also, to assess therelationship between NGAL and the severity of renal impairment assessed by microalbuminuria. With the present study, we aimed at evaluating the levels of NGAL, in serum (sNGAL) and urine (uNGAL) from a cohort study of 33 patients with type 2 diabetes mellitus categorized according to Albumin creatinine ratio (ACR) into two groups (normoalbuminuria and microalbuminuria). They were compared with age and sex matched controls. All groups showed increased NGAL values with respect to controls; interestingly, increased NGAL levels were already found in diabetic patients without early signs of glomerular damage (normoalbuminuric). Both sNGAL and uNGAL increased in parallel with the severity of renal disease, reaching higher levels in patients with microalbuminuria. The assessment of pearson coefficient evidenced that uNGAL was positively correlated to microalbumin (r=0.54, p<0.001), ACR (r= 0.72, p<0.00001) and negatively correlated to serum albumin(r==-0.45, p< 0.001).The present results suggest that normal-range albuminuria does not exclude diabetic nephropathy defined as increased sNGAL and uNGAL concentration. NGAL measurement can be more sensitive than microalbuminuria and may become a useful tool for evaluating renal involvement as well as for the early diagnosis of nephropathy in type 2 diabetic patients.

9/39 FORMULATION AND EVALUATION OF COLON TARGETED TABLETS CONTAINING SECNIDAZOLE

Amal S. M. Abu El-Enin

Department of Pharmaceutics and Industrial Pharmacy, Faculty of Pharmacy,

Al-Azhar University, Nasr City, Cairo, Egypt

The purpose of this research was to develop compression coated tablets of secnidazole for colonic delivery based on the principle of microbial degradation. Further, it was aimed to investigate the effect of combinations of selected polysaccharide polymers (guar gum, xanthan gum, almond gum and chitosan) on release profile of secnidazole. Thus present investigation is focused on formulation of secnidazole compression coating tablets to deliver the drug specifically to colon and providing better therapeutic efficacy than the conventional dosage form containing secnidazole.Secnidazole is 1-[2-Hdroxy]-2-methyl-5-nitroimidazole, used as antiprotozoal, antiamoebic and for the treatment of colonic infections.Core tablet containing secnidazole was compression coated with various proportions of guar gum, xanthan gum, almond gum and chitosan either alone or in combinations. Drug release studies were performed in simulated gastric fluid (SGF, pH 1.2) for 2 h followed by simulated intestinal fluid (SIF, pH 7.4) up to 24 h. Secnidazole release from the prepared formulations was dependent on boththe type and concentration of polymer used in the formulation. Twelve formulations were prepared by the direct compression method. The tablets were subjected to thickness, weight variation test, drug content, hardness, friability, and in vitro release studies. All the tablet formulations showed acceptable pharmacotechnical properties and complied with specifications for tested parameters. The results of dissolution studies indicated that formulation F6 (coated with mixture of guar gum and chitosan), F11 and F12 (coated with mixture of guar gum, xanthan gum and chitosan), could release the maximum amount of the drug into the colon, so they were selected for the stability study. The colon targeted tablets of secnidazole showed no significant changes either in physical appearance or drug content after stability studies conducted at 40°C / 75 % RH for 3 months. When the dissolution study was conducted in the simulated physiological environment as described under experimental sections, no significantchange in dissolution pattern was observed in the secnidazole release.

10/39 DETECTION OF M235T AND T174M POLYMORPHISMS IN ANGIOTENSINOGEN GENE AMONG EGYPTIAN PATEINTS WITH ESSENTIAL HYPERTENSION

Ola S. Ali, Yehia Z. Gad*, Mona L. Essawi*, Shereen S. Elshaer and Amal A. Mustafa

Biochemistry Department, Faculty of Pharmacy (Girls), Al-Azhar University.

*Department of Medical Molecular Genetics, National Research Center

Genetic factors contribute to the pathogenesis of essential hypertension (EH). Polymorphisms of genes encoding components of renin-angiotensin system, including angiotensinogen (AGT) were postulated to play a role in the development of EH.As a pilot study, we studied 50 patients with EH and 30 healthy individuals to assess the association of M235T and T174M AGT polymorphisms and EH among Egyptian cases. Genomic DNA samples were extracted from cases and controls and consequently amplified by polymerase chain reaction (PCR). The M235T and T174M AGT genotypes were determined by digestion of the PCR products with PsyI and NcoI, respectively.The frequencies of M235T genotypes in EH patients were MM 0.22, MT 0.62, and TT 0.16, not significantly different (p=0.471) from that of the controls (MM 0.233, MT 0.70, and TT 0.067). For the T174M polymorphism, the genotype frequencies in hypertensives were TT 0.82, TM 0.18, not significantly different (P=0. 584) from that of the controls (TT 0.86, TM 0.13).This study shows that M235T and T174M variants are not associated with essential hypertension in the studied Egyptian sample. However, the homozygous 235TT genotype tends to be more frequent in hypertensives than in controls.

11/39 THREE-DIMENSIONAL POWER DOPPLER SONOGRAPHY IN EARLY DIAGNOSIS OF FETAL GROWTH RESTRICTION

Ahmed M. Rammah

International Islamic Centre of Population Studies & Research, Al-Azhar University.

Early detection and management of fetal growth restriction (FGR) is very important in prenatal care. Three-dimensional power Doppler sonography assessment of fetal middle cerebral (MCA) & Umbilical (UA) arteries is very essential in pregnancies compromised with by fetal growth restriction (FGR). Fetal Renalvolume (RV) in addition to fetal biometrical dimensions might be one of the important parameters   for the prediction of fetal growth restriction (IUGR).This is a prospective   case- control observational study carried out at Al Adan Hospital, Al Ahmadi, Kuwait State. We used three-dimensional power Doppler sonography to assess the efficacy of fetal Blood Flow Indices, Biometrical   Dimensions and Fetal Renal volume in predicting FGR. The study was carried out on One hundred pregnant ladies compromised by growth restriction in addition to One hundred women with normal pregnancy taken as a control group.For each case Three-dimensional power Doppler sonography study   of blood flow in MCA & UA, renal volume assessment in addition to Fetal biometric diameters .Mean biometrical dimensions, Mean Pulsatility index (PI), Resistance index (RI) of MCA & UA and   Mean right and left fetal renal volumes. T he PI & RI of MCA was significantly lower in 12% of the growth restriction fetui, while The PI & RI of the UA was significantly increased in 34%of the same group. Also, Renal volume in the growth restricted fetui was significantly smaller than normally grown fetui by 43%.Growth restricted fetui appear to be potentially hypoxic & changes in blood flow resistance in UA antedate that of MCA. Fetal Growth Restriction has an association with reduction of the fetal renal volume. This study supports the hypothesis that growth restriction may be directly related to oligonephropathy and liability to develop hypertension in later life.

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