Vol. 27, July, 2008.

1/27 EVALUTATION OF IBUPROFEN EFFECT ON THE PRIMARY ANTIBODY RESPONSE IN VITRO

Saleh A. Bahashwan and *Ali A. Abdelrahman

Pharmacy Department, College of Health Sciences & *Medical Microbiology and Immunology Department, College of Medicine, Taibah University, Medina Monawarh, Saudi Arabia

Ibuprofen is a widely used anti-inflammatory and analgesic drug of special interest. Its effect on the primary immune system is not established. The primary immune response plays an important role in body defense mechanism against bacteria, viruses and cancer. Immunomodulation by pharmacological or chemical agents may alter such a defense mechanism. This study was conducted to characterize and determine the effect of Ibuprofen on the murine primary immune response (as immunoglobulin M, IgM) in vitro. The primary anti sheep erythrocyte was generated by C57BL/6 splenocytes (l x107), cultured for five days in media supplemented with 20% fetal calf serum and 2 x l06 sheep red blood cells (SRBC). Splenocytes were then harvested and the Jerne plaque assay and complement fixation tests were used to determine the numbers of plaque forming cells (PFC) and 1gM titer, respectively. Exposure of splenocytes to Ibuprofen in a concentration of (5 x10-8 to 6 x10-7 M) has resulted in augmentation of the antibody PFC (17% - 59%) and IgM titer response in a dose-dependant manner against SRBC in vitro. Ibuprofen has a significant immunostimulation in comparison to the control. However, higher concentration of Ibuprofen (1 x10-3 to 1 x10-5 M) have provoked suppression of the control antibody PFC and IgM titer response (95% - 29%) associated with 26% maximum decrease in cell viability and yield as confirmed by trypan blue exclusion. Ibuprofen at 1 x10-6 M did not show any immunostimulation without affecting cell viability and yield. We concluded that the Ibuprofen has a dual effect on the antibody response. i.e, augmentation of the primary immune response in vitro mainly occurs at 1x 10-7 M concentration of Ibuprofen.

2/27 BIOMIMETIC COATING OF CORALS TREATED WITH SODIUM HYPOCHLORITE COMPARED TO

CALCIUM HYPOCHLORITE

*Wafa I. Abdel-Fattah, *Gehan T. El-Bassyouni and **Hadeer Ibrahim

*Biomaterials Department National Research Center, Cairo, Egypt.

** Biophysics Department, Ain-Shams University, Cairo, Egypt.

The present work was concerned with treating corals with different concentrations of sodium hypochlorite between 3 and 10 weight % at several time intervals up to 3 hours at room temperature. The optimum conditions of bleaching were compared to similar treatment with calcium hypochlorite at equal hypochlorite (OCl) concentration. The changes in the treated coral samples were followed up through measuring porosity %. Samples obtained at two hours treatment with Na-or Ca-hypochlorite were selected for chemical analysis in comparison to the control untreated sample. Atomic absorption was used to determine calcium and strontium contents in the samples. X-ray diffraction analysis (XRD) was complemented by FT-IR for all samples. Thermal analysis (DTA and TGA) differentiated the effect the of treatment. Hg-porosimeter was used to assess interconnectivity. Biomimetic coating was introduced by immersing samples in solutions having chemical composition similar to a cellular fluids i.e. simulated body fluids (SBF) up to 72 hours and measuring Ca (calcium) Pi (Inorganic phosphorous) and strontium ions biochemically. SEM was used to assess architectural features pre- and after immersion in SBF. Results of apparent porosity% upon chemical treatment proved that 3% of sodium hypochlorite at 2 hours immersion was sufficient to introduce intermediate porosity values without further re-precipitation. Equivalent concentration of Ca-hypochlorite was more effective as porosity % and having higher value compared to Na-hypochlorite. DTA analysis recorded only endo-thermic reactions without any exothermic ones upon Ca (OCl)2 treatment due to leaching. XRD and FT-IR data successfully detected the metastable aragonite structure with limited isomorphous substitution of strontium in the original corals. Hg-porosimeter proved that sodium treated sample has minimum porosity, pore % and pore. The samples   treated   with Ca-hypochlorite showed more porosity than the control and the Na-hypochlorite treated one. In all cases density and skeletal density of control were higher. SEM morphological features coincided with porosimeteric data. The homogeneous and heterogeneous hydroxyapatite layer formed detected biochemically and supported by SEM predicted the future biolayer formation upon bone grafting.

3/27 BACTERIAL LIPOPOLYSACCHARIDE-INDUCED OXIDATIVE STRESS IN ADULT RAT SERTOLI CELLS IN VITRO

Hamdy A.A. Aly and *Hany A. El-Shemy

Department of Pharmacology and Toxicology, Faculty of Pharmacy, Al-Azhar University, Nasr City, Cairo, Egypt.

*Department of Biochemistry, Faculty of Agriculture, Cairo University, Giza, Egypt.

Inflammatory reactions result from microbial infections, localized as well as systemic, are reported to cause transitive or permanent male infertility. The cellular mechanism underlying the inhibitory effect of microbial infection on spermatogenesis is not fully investigated. However, there is an evidence that spermatogenesis is affected during bacterial lipopolysaccharide (LPS)-induced acute inflammation. The study used an LPS-Sertoli cell model to investigate the potential oxidative stress and toxicity of LPS in primary culture of adult rat Sertoli cells. Sertoli cells were established and incubated with different concentrations of LPS (5, 10 or 20 µg/ml) for 6, 12 and 24 h. Lipid peroxidation (LPO) and hydrogen peroxide (H2O2) production, along with superoxide dismutase (SOD), glutathione peroxidase (GPx), catalse (CAT), glutathione reductase (GR), glutathione-S-transferase (GST), γ-glutamyl transpeptidase (γ-GT) and reduced glutathione (GSH) were measured in these cells. LPO as well as H2O2 production were significantly increased while antioxidant enzyme activity, γ-GT and reduced GSH level were significantly depressed in a dose and time-dependent manner at all incubation periods with 10 & 20 µg/ml LPS. Incubation of Sertoli cells with 5 µg/ml LPS for 12 & 24 h significantly increased oxidative status while significantly decreased the antioxidant enzymes, γ-GT and GSH level. On the other hand, the oxidative and antioxidant status and γ-GT did not show any significant change in treated Sertoli cells with 5 µg/ml LPS for 6 h. The results concluded that LPS induces oxidative stress in Sertoli cells and adversely affects Sertoli cell functions.


4/27 NEW BIOLOGICALLY ACTIVE BIANTHRAQUINOID C-GLYCOSIDES FROM ASPHODELUS MICROCARPUS SALZM. ET VIVI

Abd El-Salam I. Mohammad and Ehab A. Ragab

Pharmacognosy Department, Faculty of Pharmacy, Al-Azhar University, Nasr City, Cairo, Egypt,

In this paper we report the isolation and structure elucidation of two new bianthraquinoid C-glycosides; asphodeoloside A (asphodelin-10`-oxanthrone-10`-β-D-xylopyranoside) [1] and microcarpinoside A (chrysalodin-10`-oxanthrone-10`-β-D-glucopyranoside) [2], from the tubers of Asphodelus microcarpus. The ethyl acetate fraction as well as the isolated compounds [1] and [2] showed a promising activity in the treatment of psoriasis of the scalp.

 

5/27 SYNTHESIS AND ANTIMICROBIAL ACTIVITY OF CERTAIN THIOPHENE-2-SULPHONYL-AMINO ACID DERIVATIVES

Ahmed M. El-Naggar*, Tarek M.A. Ibrahim* and Karam A. El-Sharkawy**

*Chemistry Department, Faculty of Science, Al- Azhar University, Nasr City, Egypt.

**Faculty of Pharmacy , Organic Chemistry Department, MSA University, El-Wahat Road, 6 October City, Giza, Egypt

The synthesis of different thiophene-2-(sulphonyl-DL-methionyl, p -aminobenzoyl and p-amino benzoyl DL-methionoyl- amino acid methyl esters) and corresponding amino acid hydrazides (2-29) are described. Some of the synthesized compounds were found to possess antimicrobial activities toward different microorganisms included gram-positive, gram-negative microorganisms Microccus luteuss, Bacillus subtilis, Pesudomonas aeruginosa and Escherichia coli, and the fungi Candida utilis.

6/27 SEROLOGICAL AND BACTERIOLOGICAL INVESTIGATIONS AMONG CHILDREN SUFFERING FROM RHEUMATIC FEVER

Hamido M. Hefni*, Ahmed M. El – Sersawy**, Wafaa N. El – Tayeb***

and Wael Tawakkol****

*Department of Microbiology and Immunology, Faculty of Pharmacy (Boys),

Al-Azhar University, Cairo, Egypt.

**Department of the Orthopedic Surgery, El Sahel Teaching Hospital, Cairo, Egypt.

***Department of Microbiology and Immunology, Faculty of Pharmacy, Misr International University, Cairo, Egypt.

****Department of Microbiology and Immunology, College of Pharmacy, Misr University for Sciences and Technology, Cairo, Egypt.

This study was deemed necessary in order to define the serological, hematological and bacteriological tests which help in increasing the sensitivity and specificity of rheumatic fever diagnosing. One hundred and eight patients and volunteers were selected from Orthopedic Surgery Department at El- Sahel Teaching Hospital (Cairo-Egypt), during the period from March 2006 to November 2007. Their ages ranged from 4 to 15 years old. Patients were divided into four groups. The first group (22 patients) who were hospitalized after clinical diagnosis as acute rheumatic fever patients. The second group (28 patients) and the third group (30 patients) had history of rheumatic fever. The second group did not take long acting penicillin (L. A. P.), whereas, the third group took L.A.P. once a month, as a prophylactic treatment.The different groups of rheumatic fever patients were compared with healthy control group (28 individuals) with respect to the incidence of AntistreptolysinO (ASO), Antistreptococcal Hyaluronidase (ASH), Antistreptokinase (ASK), antistreptodornase B (anti-DNaseB), Erythrocyte Sedimentation Rate ( ESR), Hemoglobin content (Hb), Red blood cells (RBCs) count, total and differential leucocytes count as well as bacteriological detection of B- hemolytic streptococcal infection. The incidence of positive tests of ASO, ASH, ASK, and anti-DNaseB was decreasing between all groups in the following pattern I > II > III> IV. In addition, the mean values of all hematological parameters, except for Hb. and RBCs count, were following the above mentioned pattern. Both serological and hematological results of group III patients did not show any significant differences from those of healthy group IV. The sensitivity of the above serological tests in diagnosis of rheumatic fever was increasing in the following pattern anti-DNaseB > ASO > ASH > ASK.      

7/27 SYNTHESIS AND ANTIMICROBIAL ACTIVITY OF THIAZOLE-4-ONE DERIVATIVES

Karam A. El-Sharkawy

Organic Chemistry Department, Faculty of Pharmacy, October University for Modern Sciences and Arts "MSA", El-Wahat Road , 6 October City, Egypt.

The synthesis of 2-cyanomethyl-thiazole-4-one obtained via the reaction of malononitrile with thioglycolic acid followed by the reaction with either acetoacetanlide and 4-chloro acetoacetanlide to produce compounds (2,3) which were reacted with either aromatic aldehyde or diazonium chloride derivatives to produce compounds (10-21, 28-33) . Some of the synthesized compounds are found to possess antimicrobial activities towards different microorganisms included two strains of Gram positive bacteria (Bacillus subtilis CECT 498 and Bacillus cereus CECT 148), one strain of Gram negative bacteria (Escherichia coli ECT 101 ) and Candida albicans CECT 1394 as a representative species of fungi was investigated.

8/27 ETIOLOGY AND ANTIBIOTIC RESISTANCE PATTERNS OF HOSPITAL-ACQUIRED URINARY TRACT INFECTIONS IN AT MISR UNIVERSITY FOR SCIENCE AND TECHNOLOGY EDUCATIONAL HOSPITAL.

Mahmoud Abd-Elati*, Wael Tawakkol*, Mohamed M.M. Ibrahim**

and Abd El-Gawad Hashim***

*Department of Microbiology and Immunology, College of Pharmacy and **College of Biotechnology, Misr University for Science and Technology.

***Microbiology and Immunology Department, Faculty of Pharmacy, Cairo University, Cairo, Egypt.

Bacterial pathogens responsible for urinary tract infections (UTIs) over a period of 16 months in a University public Hospital were identified. The level of sensitivity to five of the most commonly used antimicrobial agents was determined. Demographic distribution of pathogens was determined and related resistance and empirical therapy consecutive urine samples were collected from eighty six in patients and analyzed by standard procedure. Among eighty six isolates, Escherichia coli was the commonest cause of nosocomial urinary tract infections (UTIs). It was responsible for (50%) of infections, followed by Klebsiella spp (30%), Pseudomonas aeruginosa (10.5%), Proteus mirabilis (4.5%), Acinetobacter spp. (1.5%) and Staphylococcus spp. (3.5%). Nitrofurantoin was found effective in vitro on 64% of isolates followed by ceftazidime 58%, ciprofloxacin 55% and norfloxacin 53%, while amoxicillin was the least effective one 15%. Eleven isolates was suspected to antimicrobial susceptibility investigation to other agents used in treatment of UTI , these antibiotics were methicillin, cephotaxime, ceftriaxone, cefepime, cephamycin, cephoxitin, aztreonam, Imipenem, meropenem, gatifloxacin, gentamicin; tobramycin and amikacin. With the exception of Imipenem and meropenem, all eleven isolates showed resistance to all b– lactams antibiotics. Plasmid profile analysis of these isolates revealed that all of them harbor at least one plasmid. Plasmid curing revealed the role of plasmid in mediating antimicrobials resistance of those isolates.

9/27 THE USE OF LOW-FREQUENCY ULTRASONIC TO ENHANCEMENT OF ANTIBIOTICS ACTION AGAINST ESCHERICHIA COLI

Abdel Nasser A. EL-Moghazy

Department of Microbiology and Immunology, Faculty of Pharmacy,

Al-Azhar University and Biotechnology Center, EIPICO

Ultrasound has been investigated as an adjuvant to antimicrobial therapy, targeted at both planktonic and biofilm organisms. The effect of the aminoglycosides antibiotics upon planktonic culture of E.coli with and without an application ultrasound was studied. The aminoglycosides, genatmicin, tobramycin and amikacin, were chosen in this study because of its wide use against Gram-negative bacteria. The values of MIC of gentamicin, tobramycin and amikacin, were 8ug/ml, 8ug/ml, 10ug/ml respectively. The ultrasound was applied at level that had no inhibitory effect in order to prevent killing of bacterial cells by the used ultrasonic. Measurements of viability below the minimum inhibitory concentration of the used antibiotics showed that a simultaneous application of ultrasound and antibiotics significantly increased the effectiveness of the selected antibiotics. Bacterial viability was reduced several orders of magnitude when ultrasound were combined with gentamicin. A significant enhancement of the effect of tobramycin against growth of E.coli was observed in combination with ultrasound comparing to the effect of tobramycin alone. The combination of ultrasound and amikacin showed enhanced bactericidal activity, producing greater than 2 times more inhibition of growth than antibiotic alone after 6 h of exposure. E. coli susceptibility was also shown to vary with growth conditions, where by cells cultivated either on agar or harvested from the stationary phase of liquid culture were significantly more resistant to combination of antibiotics and ultrasound than an equivalent population obtained from the exponential phase of liquid growth. These results may have application in the treatment of bacterial infections normally resistant to some antibiotics and may be useful for expanding the number of drugs available for treating infections by rendering bacteria susceptible to normally ineffective drugs.

 

10/27 BIOLOGICAL ACTIVITY OF DIFFERENT FRACTIONS OF THE METHANOLIC EXTRACTS OF THE EGYPTIAN ANTIFERTILITY DRUGS HIBISCUS ROSA-SINENSIS LINN. AND RUTA GRAVEOLENS LINN.

Mohamed S. Hifnawy a, Shadia M. Azzam a, Iman B. Shaheed b, Sally M. Abdel-Latifa

a Pharmacognosy Department, Faculty of Pharmacy, Cairo University, Cairo, Egypt

b Pathology Department, Faculty of Veterinary Medicine, Cairo University,

Giza, Egypt

The methanolic extracts ofHibiscus rosa-sinensis Linn. flowers (Malvaceae) and Ruta graveolens Linn. leaves (Rutaceae), obtained after successive extraction with petroleum ether and methylene chloride, demonstrated potent antifertility activities in vivo and in vitro respectively in our previous study (Hifnawy et al., 2008). The ethyl acetate, butanol and water soluble fractions of each of the methanolic extracts were similarly tested in female rats at the same effective dose levels as the primary extracts either postcoitally in vivo (Hibiscus rosa-sinensis) or on the isolated uterine smooth muscle in vitro (Ruta graveolens) in order to determine the most potent extract incorporating the active ingredient(s) responsible for antifertility activity. While the water soluble fraction of Hibiscus rosa-sinensismethanolic extract showed the highest anti-implantation activity (60 %), the lowest estradiol serum levels and the most severe pathological lesions in the ovaries among the three fractions, the butanol soluble fraction of the methanolic extract ofRuta graveolens demonstrated the most potent uterotonic activity and in a dose dependent manner. Possible significances of such findings are discussed.


11/27 PHYTOCHEMICAL AND BIOLOGICAL INVESTIGATION OF COUMARIN CONTENT OF PELARGONIUM FRAGRANS - WILLD. GROWN IN EGYPT

Khaled M. Meselhy

Department of Pharmacognosy and Medicinal Plants, Faculty of Pharmacy,

Cairo University, Cairo, Egypt

Three coumarins viz; C1 (Umbelliferone), C2 (Umckalin) and novel coumarin glycoside C3, were isolated from Pelargonium fragrans Willd. Isolated compounds were identified on the basis of their physico-chemical, UV spectral data, 1H-NMR, 13C-NMR and EIMS. These compounds were isolated for the first time from the plant. Significant antiwormal effect on fresh earthworms (Allolobophora caliginosa), remarkable antioxidant activities and strong antifungal activity were recorded on the extract, fractions and isolated compounds.


12/27 DESIGN AND EVALUATION OF CLOTRIMAZOLE BIOADHESIVE VAGINAL TABLETS

Afaf A. Ramadan

Department of Pharmaceutics and Industrial Pharmacy, Faculty of Pharmacy, Al-Azhar University, Nasr City, Cairo, Egypt.

Clotrimazole is an imidazole derivative which is broad spectrum antimycotic agent against pathogenic dermatophytes, yeasts and several species of Candida, Trichophyton, Microsporum, Epidermophyton and Malassezia.oral bioavailability of clotrimazole is poor and it has a high incidence of gastrointestinal disorders and neurological reactions. However, clotrimazole is known to be very effective locally and causes no major side effects. So, it was formulated as vaginal bioadhesive tablets using different polymers by direct compression technique. Physical properties of the tablets and the in vitro release of the drug were studied using USP basket dissolution apparatus. The results obtained revealed that, for the preparation of effervescent vaginal bioadhesive tablets, the most suitable concentrations of the polymers were found to be 30%, 10% and 2% for HPMC, CMC Na and carbopol 934, respectively. Physical standards of the prepared tablets were found to be significantly influenced by the polymer concentration. Increasing polymer concentration resulted in an increase in both hardness and time required for complete disintegration. In Vitro release rate of clotrimazole from the prepared vaginal bioadhesive tablets was highly affected by the concentration of the polymer used. Increasing the polymer concentration significantly decrease the drug release rate. The type of the bioadhesive polymer was found to affect the in vitro drug release. At the same polymer concentration, the retarding effect of the investigated polymers could be arranged as: carbopol 934 > CMC Na > HPMC. Kinetic analysis of the release data showed that all the investigated tablets except the formula no. 4 released clotrimazole in a zero – order mechanism. Formula no. 4 which is formulated using 20% CMC Na showed non – Fickian diffusion mechanism. Storage for six months at room temperature for the most suitable formula was found to have a little effect on the physical parameters of the tablets, drug content as well as in vitro drug release. All selected tablets complied with the pharmacopieal requirements at the end of the storage period.

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