Vol. 16, November, 2004.

1/16 BIOFILMS FORMATION BY STAPHYLOCOCCUS EPIDERMIDIS: MODULATION BY FLUOROQUINOLONES

M. Yassien and N. Khardori*

Faculty of Pharmacy, King abdul Aziz Universaity, Jeddah, Saudi Arabia and *Department of Medicine, Division of Infectious Diseases, Southern Illinois University School of Medicine, Springfield, Illinois.

The interaction between pefloxacin, ciprofloxacin, norfloxacin, ofloxacin, and levofloxacin and biofilms formed by Staphylococcus epidermidis (20 clinical isolates) was studied. In the presence of 1/2 MIC, 1/4 MIC and 1/8 MIC, the optical density of the biofilms was reduced to 22-24%, 44-52% and 65-74% of the controls, respectively. Treatment of preformed biofilms with fluoroquinolones in concentrations ranging from 12.5 μg/ml to 400 μg/ml, caused reduction in the optical density of the adherent biofilms to 45-77% of the control. In an in vitro model of vascular catheter colonization, subinhibitory concentrations (1/2, 1/4, or 1/8 MIC) of fluoroquinolones reduced the number of adherent bacteria to 24-28%, 48-55% and 58-76% of the controls, respectively. The vascular catheter segments precolonized with Staphylococcus epidermidis for 24 hours and exposed to the fluoroquinolones in 4-8 times MIC (50 μg/ml) for 2 hours showed less than 8% growth of adherent cells compared to the controls. No adherent organisms were cultured in the presence of 8-16 times MIC (100 μg/ml). The activity of pefloxacin in reducing the bacterial adherence and eradicating the preformed biofilms was demonstrated by scanning electron microscope. The effect of the fluoroquinolones on the adherence of Staphylococcus epidermidis to the surface of human epithelial ( Hep-2) cells was also studied. In the presence of subinhibitory concentration (1/2, 1/4, and 1/8 MIC), the range of the number of the adherent viable cells was reduced to 39-48%, 46-61%, and 62-75 % of the control, respectively. Treatment of Hep-2 cells, preattached with bacterial cells, with the tested fluoroquinolones at concentrations of 5, 10, and 20 μg/ml decreased the number of the adherent cells to 64-71%, 53-61%, and 29-37% of the control, respectively. These data show that subinhibitory concentrations of pefloxacin, ciprofloxacin, norfloxacin, ofloxacin, and levofloxacin inhibit the adherence of Staphylococcus epidermidis to the surfaces of plastics, vascular catheters, and human epithelial (Hep-2 ) cells. In addition, higher concentrations of fluoroquinolones were able to eradicate the preformed biofilms of Staphylococcus epidermidis.

2/16 ADIPOCYTOKINES AND INSULEN RESISTANCERELATIONSHIPS IN CHILDHOOD OBESITY

A. Abdel-Lateef

Radiological Health Research Department, National Centerfor Radiation Research and Technology, Cairo, Egypt.

This study aimed to find out the relationship between obesity degree in children and tumor necrosis factor-alpha (TNF- µ ) and adiponectin. Also, to estimate whether these adipocytokines are as closely correlated with fasting insulin and insulin resistance as shown for adults. The present study was carried out on 29 obese children (16 boys and 13 girls). The diagnosis of obesity was based on the presence of body mass index (BMI) > 95Th percentile for age and gender. Their mean age was 10.03 ± 1.96 year and their mean BMI was 24.02 ± 4.28 kg / m2. All children were tanner stage I. Children with secondary obesity were excluded. They were chosen from the outpatient clinic of Ain Shams Pediatric Hospital. Another 15 healthy children with matched age and sex served as control group. All the studied groups were subjected to full clinical examination, anthropometric measurements and estimation of: fasting blood glucose, fasting insulin and fasting plasma adiponectin by radioimmunoassay, TNF -µ by ELISA. Fasting plasma glucose and fasting insulin levels were used to calculate a factor known as homeostasis model assessment for insulin resistance (HOMA – IR). This study showed that gender had no effect on biochemical analyses of obese children. So, the results of boys and girls were analyzed together. Obese children showed significant increase in: body weight, BMI, fasting insulin, HOMA – IR and TNF-µ - than control. Adiponectin showed significant decrease in obese children with p < 0.001 when compared to control. These alterations became pronounced by the increase of the degree of obesity. It was the adiponectin that significantly decreased in mild obesity with p < 0.05. It may be an evidence that adiponectin is an early mediator of obesity- induced insulin resistance. On correlating these biochemical parameters with each other, it was found that: both of obesity degree and TNF- µ had markedly correlated with insulin and HOMA – IR with p < 0.0001 in all cases. Meanwhile, adiponectin correlated inversely with obesity, TNF -µ insulin and HOMA- IR with p <0.0001 in all tests. So different from TNF-µ, adiponectin has an endogenous insulin sensitizing effects. It was concluded that measurement of adipocytokines might become a part of the standard evaluation of the obese child and help to manage those overweight children who may be at high risk for the development of type 2 diabetes later in life.

 

 

3/16 STUDY OF CARBOHYDRATES, LIPIDS, PROTEIN, FLAVONOIDS, VITAMIN C AND BIOLOGICAL ACTIVITY OF PSIDIUM GUAJAVA L. GROWING IN EGYPT

S.M. Abdel Wahab; M.S. Hifnawy; S.M. Azzam, H.M. El Gohary and *M. IsaK

Pharmacognosy Department, Faculty of Pharmacy, Cairo University.

* Pharmacology Department, Faculty of Medicine, Cairo University.

Phytochemical investigation of leaves, fruits and seeds of Psidium guajava L. revealed the presence of carbohydrates and/or glycosides, tannins, saponins, flavonoids, sterols and/or triterpenes and essential oil. GLC study of the lipoidal matter in leaves and seeds revealed that the major compounds were: hydrocarbons; n-tricosane (7.34%) and n-triacontane (11.01), steroids; ß-Sitosterol (24.7 and 31.01%) and fatty acids; palmitic acid (39.67 and 49.43%) in the leaves and seeds respectively. Free sugars, pectin, vitamin C, protein and amino acids were investigated in fruits. Their isolation, identification and quantification were achieved using HPLC and titremetric methods. Tannin content of the seeds, fruits and leaves was determined using both gravimetric and redox methods in addition to the crude fibre content of the seeds. Study of flavonoid content of leaves resulted in isolation of five compounds; quercetin, Kampferol, quercetrin, apigenin 7-O-glucoside and luteolin 7-O-glucoside. Total flavonoids was estimated 0.57% calculated as quercetin. Toxicological study indicated that leaves were safe for use in a concentration up to 2.5g/kg body weight (aqueous extract). Pharmacological study revealed that essential oil, aqueous, alcoholic and ethyl acetate extracts of leaves exhibited a significant muscle relaxant activity on the smooth muscle especially that of the trachea, as well as, a significant anti-inflammatory activity, that explains the use of guajava leaves in the Egyptian folk-medicine for treatment of cough.

 

 

4/16 EVALUATION OF THE MOLLUSCICIDAL AND IN VITRO SCHISTOSOMICIDAL ACTIVITY OF BUTANOL EXTRACT OF THE PLANT AGAVE LOPHANTHA

A. Abdel Kader*, F. Ramzy** and A. Tantawy*

*: Environmental Researches and Malacology Department   **: Parasitology Department, Theodor Bilharz Research Institute, P.O.Box:30 Imbaba, Egypt

Butanol extract of the leaves of Agave lophantha which is belonging to family Agavaceae from Egyptian flora was tested for molluscicidal and schistosomicidal activities. The results show that the LC50 values were 23.5, 35, 20 and 20 ppm against Biomphalaria alexandrina, B.glabrata, B.truncatus and L.natalensis snails, respectively. Low concentrations (1/10 LC50) of the extract of A.lophantha caused 100% mortality among the exposed B.alexandrina, B.truncatus and L.natalensis and 90% mortality for B.glabrata snails after 4 weeks of exposure to low concentrations. The egg production was inhibited (100%) markedly at the end of the exposure period. It caused 14.6 (after 3 weeks) and 31.3% (after 2 weeks) in ASAT and ALAT enzymes in the hemolymph of B.alexandrina snails, 40.5% decline in total protein after 3 weeks, 86.7% decrease in albumin at the end of exposure and 114.1% elevation in total lipids at the end of 4th. week reflecting the damages caused by the exposure to low concentration of plant extract. A considerable in vitro effect of the butanol extract of A.lophantha on Schistosoma mansoni worms where the LC50 was 14.5 ppm, compared to praziquantel (LC50: 0.287 ppm. 100% of the exposed cercariae and miracidia were killed after 6 and 24 hours of exposure to the lowest concentration (1 ppm).

 

5/16 DETECTION OF HCV RNA IN THE SALIVA OF CHRONIC HEPATITIS C PATIENTS

  1. ¨ and A. Abdel Kader¨

*Microbiology Department, Faculty of Medicine for Girls, Al-Azhar University and ¨Liver Unit, Ahmed Maher Teaching Hospital

Fifty Egyptian chronic hepatitis C patients and 15 healthy control subjects were included in the present study. Serum and saliva samples were taken from them. Serum samples were subjected to testing for anti-HCV by ELISA test and for HCV RNA by RT-nested PCR. Serum ALT and AST levels were also determined. Saliva samples were also tested for HCV RNA by RT-nested PCR and for the presence of occult blood by benzidine test as a preliminary test and by Teichmann test as a confirmatory test. It was found that all the 50 chronic hepatitis C patients had anti-HCV in serum, while 47 of them (94%) had HCV RNA in serum. Twenty-eight of the 50 patients (56%) were found to have HCV RNA in saliva; all of them were viremic as they had detectable HCV RNA in serum. The remaining 22 patients (44%) were negative for HCV RNA in saliva, including 19 viremic patients and 3 non viremic patients. No correlation was found between the presence of occult blood, indicating blood contamination of saliva, and detection of HCV RNA in saliva, as occult blood was detected in 13 saliva samples, 7 of them were HCV RNA – positive and 6 were HCV RNA – negative. All control subjects were negative for anti-HCV in serum and HCV RNA in serum and saliva, and no occult blood was detected in saliva. No statistically significant difference in age, sex or the duration of the disease was found between patients with and those without HCV RNA in saliva (P>0.05). But the family history of liver disease was significantly higher in patients with HCV RNA in saliva than in patients without HCV RNA in saliva (P<0.05), while a significantly higher proportion of patients without HCV RNA in saliva had received antiviral treatment, in the form of interferon and ribavirin, compared to patients with HCV RNA in saliva (P<0.05). Risk factors for HCV infection in patients with or without HCV RNA in saliva were mainly non-transfusion parenteral exposure, and to a lesser extent blood transfusion. However, a relatively high proportion of patients of both groups had no identified risk factors for HCV infection. No significant difference in risk factors, whether these factors were identified or not, was found between patients with and those without HCV RNA in saliva (P>0.05). On the other hand, serum ALT and AST levels were significantly higher in patients with HCV RNA in saliva compared to patients without HCV RNA in saliva (P<0.05). Detection of HCV RNA in the saliva of a high proportion of viremic chronic hepatitis C patients suggests that salivary transmission should not be excluded when considering non parenteral routes of transmission of HCV. Further studies are required to determine the mechanism by which HCV enters the saliva, and the possible transmission of HCV through exposure to saliva. Meanwhile, precautions and care should be taken when handling saliva or objects contaminated by it. The possibility of HCV transmission through saliva and other body fluids has to be fully investigated if the epidemiology of HCV is to be fully understood.

 

 

6/16 CarbamazepinE suppositories: influence of formulation on physical parameters and stability
By

A.A. Ammar and M.A. Marzouk

FROM
Department of Pharmaceutics and Industrial Pharmacy, Faculty of Pharmacy,

Al-Azhar University, Nasr City, Cairo, Egypt.

The formulation and evaluation of Carbamazepine (CBZ) rectal suppositories, as a fast release dosage form, were studied. Carbamazepine was formulated as a rectal suppository, in six formulae, adopting the molding from a melt technique. Displacement values, physical characteristics as well as CBZ release profiles from the prepared suppositories were determined. Stability study was done for all the CBZ suppository formulae. The results showed that all the investigated CBZ suppositories had acceptable physical characteristics with respect to hardness, solubility time and uniformity of drug content. The release profile from PEG bases was found to be dependent on the molecular weight of the PEG used. Accelerated stability testing obviated that all the investigated formulae met the pharmacopoeial requirements of drug content after storage for three months at 30oC and 40oC, as 94.8 to 98.6 % of un-decomposed drug was remained. The specific rate constants at the two elevated temperatures (K35 & K45) were determined by substituting in Arrhenious equation. The energy of activation (Ea), decomposition reaction constant at room temperature (K20), (t1/2) and (t90) were also determined. Differential scanning calorimetry (DSC) was used to investigate the physicochemical properties of CBZ suppositories and the effect of storage on their stability. The PEG rectal suppositories of CBZ could be prepared to be used as an alternative to oral dosage form, for fast drug release and to circumvent the first-pass metabolism.

 

 

7/16 STUDY OF SERUM LEPTIN AND INSULIN CHANGESIN HEALTHY WOMEN

M.A. El-Desouky, M.O.I. Refaie and F.M. El-Hussainy

Lab of Biochemistry, Faculty of Science, Cairo University.

Leptin and insulin Levels were measured in healthy menstruating females including young unmarried females and older cycling women during the three phases of the menstrual cycle: early follicular phase (F-phase), Mid-cycle phase (M-phase) and Mid-luteal phase (L-phase). Moreover, the relationships between leptin and LH, FSH, Testosterone and estradiol will be studied. In addition, the effect of age and Body Mass Index (BMI) on these parameters will be investigated. Subjects were classified into two groups according to age: Group (1): twenty normal healthy cycling women, with age of 18-30 years old, of cycle length of 28-32 days. Group (2): twenty normal healthy cycling women, with age 31-42 years old, of cycle length of 28-32 days, morning fasting blood samples were collected from all subjects at the early follicular phase within days 3-7 of the menstrual cycle, the mid-cycle phase within days 14-16 of the menstrual cycle and the mid-luteal phase within days 21-22 of the menstrual cycle. All cases were classified according to the BMI to lean ones whose BMI<25 and to obese whose BMI>27. Fasting leptin, insulin, LH, FSH, estradiol and testosterone were measured during the three mentioned phases. None of the women had any systemic or metabolic diseases nor taken any hormonal medications for at least one year. Our results revealed that serum LH, FSH, and estradiol concentrations were significantly increased in the mid-cycle phase and subsided thereafter in both lean and obese subjects. Testosterone concentrations were slightly increased in the mid-cycle phase than in the early follicular and mid-luteal phases of the menstrual cycle of lean and obese subjects. Leptin and insulin concentrations were increased significantly in the mid-cycle and mid-luteal phases than in the early follicular phase of the menstrual cycle mostly in lean subjects, while these differences show non-significant variations in obese subjects. On the other hand, leptin and insulin levels showed very great differences between lean and obese subjects, being higher in obese ones.Our data suggest that leptin may act centrally to alter hypothalamic and/or pituitary function and that leptin may promote ovarian function through direct actions on the ovarian follicle even in small amounts.

 

 


8/16 ULTRASTRUCTURAL CHANGES IN THE PANCREAS AND BIOCHEMICAL STUDIES OF THE HYPOGLYCEMIC EFFECT OF SOME NATURAL HERBAL PRODUCTS IN ALLOXAN DIABETIC RATS

H.H. Al Zuhair, *A.G.H. Al Zuhair, M.I. El Sayed and A.A. El Fattah

Pharmacology Department, Faculty of Pharmacy and *Department of Anatomy, Faculty of Medicine, King Saud University, Kingdom of Saudi Arabia.

A new wave of research interest in traditional practices which might be used to alleviate the symptoms of diabetes mellitus had been stimulated by renewed attention to natural therapies. Therefore, Nigella sativa “blackseed” in a mixture of equal proportion with Myrrh, Aloe, Asafetida and Olibanum frankincense for preparation of an aqueous extract were used. This extract was given as oral remedy for alloxan-induced diabetic rats, in comparison with the second generation sulphonylurea oral hypoglycemic drug “glyburide”. We investigated their effect on glucose and insulin level in blood. While specimens were collected from pancreas of the control and diabetic rats and examined for light microscopy, scanning and transmission electron microscopy. The herbal extract showed a significant hypoglycemia & increased serum insulin level either as monotherapy or in combination with glyburide at p<0.05. Histologically this herbal extract induced an obvious & effective repair and healing of the damaged pancreatic beta cells in alloxan induced diabetic rats. This result support the role of the herbal medicine.  

 

 

9/16 IN-VITRO AND IN-VIVO EVALUATION OF CONTROLLED-RELEASE TIAPROFENIC ACID TABLETS PREPARED WITH HYDROGENATED CASTOR OIL.

A. Ismail

Department of Pharmaceutics and Industrial Pharmacy, Faculty of Pharmacy, University of Al-Azhar, Assiut Branch, Assiut, Egypt

Using waxy materials, like K3 wax, several controlled release tiaprofenic acid tablets were prepared. The tablets were prepared through modification of the ratio of both hydrogenated castor oil (K3 wax) and polyethyleneglycol-6000. The tablets were orally administered to rabbits and the drug concentration in the plasma was determined by HPLC method. The plasma concentration rapidly increased after the oral administration of the aqueous solution of the plain drug, reaching a maximum 10 minutes after administration and then sharply declines. Regarding dissolution study, it was found that the release rate decreased with an increase in the amount of (K3 wax), while increasing the content of PEG 6000 caused a significant increase in the release rate. The plasma concentrations of tiaprofenic acid after oral administration of the plain tablets and four kinds of wax matrix tablets to rabbits were determined and the pharmacokinetic parameters were calculated. The ( tmax) was delayed and the ( Cmax ) was decreased with an increase in ( K3 wax) content in the tablets. The obtained results proved that the diffusion of the drug through the waxy matrix and the erosion of this matrix caused by the gastrointestinal tract (GIT) motility contributed to the in vivo dissolution mechanism. The blood levels were very low when the tablets were administered after giving food. The ( Cmax ) decreased to one -eight of that administered under fasting conditions and the plasma concentration was sustained at an extremely low level from 30 min to 7 hr after oral administration. Propantheline bromide (PB), was used in this study for the depression of (GIT) motility and the tablets were administered to rabbits pretreated with (PB). Compared with the untreated rabbits, the (Cmax ) was decreased, The (tmax) was prolonged and the (AUC) was increased 1.3 times. The plasma concentration increased gradually and the level at the 7th hour after administration was higher than that in the untreated rabbits. The concluded data suggest the benefit of using waxy matrix for the development of controlled release dosage forms of tiaprofenic acid to decrease the dosing frequency of the drug.  

 

10/16 A PRELIMINARY INVESTIGATION OF THE POSSIBLE DIURETIC ACTIVITY OF SOME JORDANIAN MEDICINAL PLANTS

H. Abdel Jalil

Pharmacology Department, Faculty of Pharmacy,

Al-Zaytoonah University of Jordan, Amman, Jordan

Lyophilized aqueous extract of the plant Sinapis Arvensis L. (Cruciferae) caused a significant increase in urine volume, potassium and chloride excretion, as compared to the control values. Lyophilized aqueous extract of Rosemary, Rosmarinus Officinalis L. (Labiatae), significantly increased urine volume, without a proportional increase in electrolyte excretion. No modifications with respect to control were observed for the urinary parameters in the pooled urine samples for rats treated with lyophilized aqueous extract of Ditrichia Graveolens L. (Compositae) by the P.O. route, recorded at 6h. Aside from a slight urinary acidifying action seen with the extract of the plant Sinapis Arvensis, no differences have been seen between the assayed, and the control groups with regard to the urinary pH.

 

 

11/16 SPASMOLYTIC AND CALCIUM ANTAGONIST ACTIVITIES OF THYMOQUINONE

R.A. M. Taha

Department of Pharmacology and Toxicology, Faculty of Pharmacy,

Al-Azhar University, Cairo.

The volatile oil of Nigella sativa seed proved to have a relaxant effect on uterine smooth muscle. There are no reports about the effect of thymoquinone (TQ) which is the main active constituent of the volatile oil of Nigella sativa seed on uterine muscles. The effect of TQ was tested on spontaneously contracting and on agonists-induced contraction in isolated rat uterus. TQ (37.5-300 µM) reduced spontaneous contraction of isolated non-pregnant rat uterus in concentration dependent manner. TQ (150 or 300 µM) reduced prostaglandin F2α, oxytocin and acetylcholine-induced uterine contractions. This was shown to shift the non-cumulative log concentration-response curves of the test agonists in concentration dependent manner to the right. This shift was accompanied by a reduction in the maximal response (Emax) of the tissue to all selected uterine stimulants and elevation of EC25. Oral administration of thymoquinone (10 or 20 mg/kg) daily for three weeks inhibited the contraction evoked by previous oxytocic agents (prostaglandin F2α, oxytocin and acetylcholine) on uterine muscles. TQ also shifted the non-cumulative log concentration-response curves of CaCl2 in a concentration dependent manner without a reduction in Emax. This effect was similar to that produced by verapamil. Besides, TQ inhibited the K+-induced contraction in a concentration dependent action. In conclusion, these results suggest that TQ-induced inhibition of the uterotonic potential of various oxytocic agents and spontaneous movements may be due to Ca2+ channels blocking action.

 

12/16 BIOAVAILABILITY AND PHARMACOKINETICS OF NICARDIPINE HCL MICROSPHERES

A.M.S. Ahmed, M.A. Marzouk and G.E. Yassin

Pharmaceutics Department, Faculty of Pharmacy, Al-Azhar University,

Nasr City, Cairo, Egypt.

The aim of this work is to study the bioavailability and the pharmacokinetic parameters of Nicardipine Hydrochloride (N HCl) microspheres prepared by emulsion-solvent evaporation technique, using Ethyl cellulose and Eudragit RS100 polymers in two drug-polymer ratios (1:1, and 1:2). The selected formulae were formula F1A with Ethylcellulose in 1:1 drug-polymer ratio (900 µm), Formula F2A, with Ethylcellulose in 1:2 drug-polymer ratio (900 µm), Formula F3A with Eudragit RS100 in 1:2 drug-polymer ratio (900 µm), Formula FB, with Ethylcellulose in 1:2 drug-polymer ratio (565 µm), and Formula FC, with Ethylcellulose in 1:2 drug-polymer ratio (407.5 µm). N HCl plasma concentration levels were analyzed using an HPLC method. The Pharmacokinetic parameters of the prepared N HCl microsphere formulae were compared with the commercially available product, namely Pelcard 50, after their oral administration to rats. The results of the pharmacokinetic parameters of the aforementioned formulae, were treated statistically, using ANOVA at P <0.05. It was found that Formula FB containing N HCl and Ethylcellulose in 1: 1 drug: polymer ratio (565 µm) was the best studied treatment as it showed: maximum Cmax, MRT, and AUC (0-24), suitable Kab and Kel and good relative bioavailability.

 

13/16 SPECTROSCOPIC INFRA RED AND PROTON MAGNETIC RESONANCE DETERMINATION OF INDAPAMIDE

M.M.F. Metwally

Analytical Chemistry Department, Faculty of Pharmacy,

Zagazig University, Egypt

Indapamide has been determined by infra red spectroscopy as well as proton magnetic resonance spectroscopy. The absorptions of bonds vibrations were found to be proportional to concentrations at wave number = 809.956 cm-1, 1044.26 cm-1, 1383.07 cm-1 and 3654.44 cm-1. The quantitative determination of indapamide by both infra-red spectroscopy and proton magnetic resonance spectroscopy were verified. The average % recovery + standard deviation is found to be 100.28 + 1.19 and 100.878 + 1.267 for infra red method and for proton magnetic resonance method respectively.

14/16 OPTICAL ISOMERS OF 3-SUBSTITUTED-5-(2-CARBOXYETHYL)-1,3,5-THIADIAZINE-2-THIONES OF POTENTIAL ANTIMICROBIAL ACTIVITY, SYNTHESIS AND SOLID STATE STABILITY

M.A. Hussein, T. Aboul-Fadl*, M.O. Ahmed**, M. Hashem***

and M. Hassan****

Dept. Pharm. Org. Chem., *Dept. Pharm. Med. Chem., **Dept. Ind. Pharmacy, Faculty of Pharmacy, ***Dept. Botany, Faculty of Science, and ****Dept. Plant Pathology, Faculty of Agriculture, Assiut University, Assiut-71526, Egypt

In an effort to study the effect of optical purity on the antimicrobial activity of the 1,3,5-Thiadiazine-2-thione moiety, twenty four 3-substituted-5-(2-carboxyethyl)- 3,4,5,6-tetrahydro-2H-1,3,5-thiadiazine-2-thione (THTT) derivatives (4-27) were synthesized by the reaction of the appropriate alkyl, cycloalkyl or aralkyl amine, carbon disulphide, formaldehyde, and the three optical forms (dl, d, and l) of alanine. The structures of the synthesized compounds (4-27) were verified on the bases of the spectral and elemental methods of analyses. Solid state stability of some of the prepared derivatives was studied using Differential Scanning Calorimetry (DSC). The measured racemic mixtures have distinctive DSC curves, which are characterized from that of the corresponding isomers. The title compounds were tested for their antibacterial activity in vitro against Gram +Ve bacteria (Bacillus cereus and Staphylococcus aureus), Gram -Ve bacteria (Serratia rodenii). In vitro antifungal activity was tested against dermatophytes (Candida albicans, and Trichophyton rubrum), saprophytes (Aspergills terreus, and Rhizopus stolonifer), phytopathogenic (Phythium debaryamum, Fusarium oxysporum, and Macrophomina phaseolina), and antagonist (Trichoderma harziamum) fungi. The investigated compounds exhibited varied inhibitory effects on the growth of the most tested microbial species. Although, there is no general pattern between the optical purity and the antimicrobial activity, chirality may have a role on the antimicroial activity of some derivatives.

15/16 PHYTOCHEMICAL AND BIOLOGICAL INVESTIGATION OF LIPID AND COUMARIN CONTENTS OF PELARGONIUM DENTICULATUM JACQ.

M.M. El-Sherei, F.S. El-Sakhawy, H.A. Kassem, K.M. Meselhy and A.A.Sleem*

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

* Pharmacology Department, National Research Center, Giza, Egypt.

The lipoidal and coumarin contents of the aerial parts and roots of Pelargonium denticulatum Jacq. were investigated. GLC analysis of the unsaponifiable matter revealed the presence of 14 components. n-Eicosane C20 (17.1%) and n-decosane C22 (16.7%) were the major hydrocarbons, while ß– sitosterol (10.1%) was the major sterol. GLC analysis of the fatty acid methyl esters revealed the presence of palmitic (24.5%) and caprylic (15.1%) as major saturated fatty acids and tridecanoleic (4.3%) and caprioleic (4.2%) as major unsaturated fatty acids. Morever, three compounds viz;α–amyrin(L1),friedelin (L2) andß–sitosterol(L3) were isolated and identified through their physico-chemical data, 1H-NMR and EIMS, as well as, through direct comparison with authentics. In addition, four coumarins viz;umbelliferone(C1),methyl-isopimpenllin(C2), scopoletin(C3) and umckalin(C4) were isolated and identified on the basis of their physico-chemical and UV spectral data, 1H-NMR, 13C-NMR and EIMS. All the previously isolated compounds were for the first time isolated and identified from the plant. The ethanolic and aqueous extracts of the aerial parts showed significant analgesic and anti-inflammatory activities without any ulcerogenic effect, in addition to a strong antipyretic, anticoagulant and antihyperglycaemic activities on the experimental animals.

 

16/16 PHYTOTHERAPEUTIC EVALUATION OF EGYPTIAN DESERT TRUFFLES

M.S. Hifnawy *, S.M. Azzam*, S.N. Soliman*, A.H. El-Ghorab**, I.B. Shaheed*** andS.M. Abdel-Latif*

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

**Flavour and Aromatic Department, National Research Center, Cairo, Egypt

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

Cairo, Egypt

The present study was carried out on two varieties of Egyptian desert truffles viz. Tirmania nivea (Desf.) Trappe (white truffles) and Terfezia claveryi (Chatin) (brown truffles). Preliminary phytochemical screening revealed the presence of volatile matter, carbohydrates and/or glycosides, sterols and/or triterpenes and traces of tannins. Eleven volatile compounds could be identified including four sulfur compounds, five alcohols, one aldehyde and one ketone. Pheromones (5-a-androst-16-en-3-a-ol and 5-a-androst-16-en-3-one) were detected in trace amounts in both species. Study of the antimicrobial activity showed a marked antibacterial activity against G+ve and G-ve bacteria with no activity against yeast or fungi, while histopathological examination revealed marked vasodilatation of blood vessels of testes and accessory glands as well as a promising effect in the treatment of eye inflammation. Also, the fungus was found to possess an androgenic effect.

 

17/16 THE HEPATOPROTECTIVE MECHANISMS OF NITRIC OXIDE IN THE OXIDATIVE LIVER INJURY

W.M. Awara, A.E. El-Sisi, M.E. El-Sayad, N.A. El-Shitany and K. El-Desouky*

Department of Pharmacology and Toxicology, Faculty of Pharmacy, *Department of Pathology, Faculty of Medicine, Tanta University, Tanta, Egypt.

The role of enhanced NO production during liver damage has not been clarified yet as the expression of NOS may be associated with both beneficial and harmful consequences. This study was designed to assess the role of NO in oxidative liver damage using acetaminophen (APAP)-induced liver injury as an experimental model. The cellular mechanisms of NO including the involvement of oxidant / antioxidant mechanisms were also investigated. In addition, the role of NO in modulating the apoptotic cell death in the liver was examined. Rats were treated with a single dose of APAP (1g/kg) I.P. either alone or in combination with L-NAME (15 mg/kg), L-arginine (100 mg/kg), and buthionine sulfoximine (BSO) as a glutathione depleting agent (1.6 g/kg). At the indicated time points, the following parameters were assessed; the plasma ALT enzyme activity, the hepatocyte contents of NO and LDH, the hepatic tissue content of lipid peroxides (measured as MDA) and reduced GSH. Histopathological examination of liver sections was also carried out. In addition, liver apoptosis was also evaluated by DNA fragmentation analysis using agarose gel electrophoresis as well as Fas expression and apoptotic index in liver sections by immunohistochemical staining (TUNEL). Pretreatment of APAP-treated rats with L-arginine resulted in protection against APAP-induced hepatic injury as presented by the significant decrease in the hepatocellular enzyme release (ALT & LDH) and attenuation of hepatocytes apoptosis and necrosis. On the other hand, L-NAME and BSO treatment significantly exacerbated the oxidative liver damage. Our results have suggested that NO may have a hepatoprotective effect against oxidative liver damage. This hepatoprotection was due to several mechanisms including attenuation of hepatic lipid peroxidation, increase the hepatic GSH contents, and/or decrease in liver Fas expression and hence decrease the apoptotic cell death and DNA damage.

 

 

18/16 FOLIC ACID, VITAMIN B12, AND HOMOCYSTEINE IN HEALTHY ACTIVE, PASSIVE AND NON SMOKER SUBJECTS.

M.A. Abbas and H.M. El-Kholy

Department of Biochemistry, Faculty of Medicine, for Girls, Al-Azhar University.

The present study was conducted in order to study the serum folic acid, vitamin B12, and their relation to serum homocysteine, in healthy three group subjects: active, passive and non smokers (controls). Forty male subjects were included in this study and were classified as the following; group I consists of ten subject as control (non smoker and not exposed to tobacco smoke), with age ranging from 21- 44 years. group II consists of fifteen subjects (passive Exposed non smokers), With age ranging from 20- 43 years. group III consists of fifteen subject (active smokers), for at least 40 cigarette per day with age ranging from 21- 49 years. The groups I and II were medical and paramedical staff in faculty of medicine Al-Azhar university. They were asked to fill in a self completion questionnaire giving details about their smoking habits. Detailed history to exclude the presence of previous or recent hepatic, renal, metabolic or gastro intestinal diseases which might affect the parameter to be investigated. Blood pressure was measured and blood samples were taken from all subjects. Levels of vitamin B12, and folate were measured in the sera from subjects of the three groups. They determined by using simul TRAC-SNB radioassy kit. Level of serum Hcy was performed using Hcy enzyme immunoassay. The results, were revealed a significant decrease in the level of serum folic acid as well as vit B12, levels in passive smokers when compared to controls and in active smokers when compared to controls and to passive smokers. A highly significant increase in the level of Hcy in passive smoker when compared to controls and in active smokers when compared to controls and to passive smokers. Also, a negative significant correlation was found between serum Hcy levels with serum folic acid and vitamin B12, in active and passive smokers. Active or passive smoking resulted in increased Hcy level which may be attributable to decreased folate and vit B12, although the mechanism was not precisely defined.

 


19/16 DETECTION OF TYPE 2- LIKE T- HELPER CELLS IN HEPATITIS C VIRUS INFECTION:

IMPLICATIONS FOR HEPATITIS C VIRUS CHRONICITY

I.A. Gaweesh* and I.A. Rewesha**

* Clinical Pathology Department, National Liver Institute Menoufeya University

** Internal Medicine Department, National Liver Institute Menoufeya University

T- lymphocytes and immunoregulatory cytokines may be important in the host response to hepatitis C virus (HCV) infection. T-helper type 1 (Th1) cytokines (IL2, IFN-) are required for host antiviral immune responses, including cytotoxic T-cell generation and natural killer cell activation, while T-helper type 2 (Th­2) cytokines (IL-4, IL-10) can inhibit the development of these effector mechanisms. In an attempt to investigate the immunopathogenesis of chronic hepatitis C virus, we analyzed Th1 cytokine (IFN-) and Th­2 cytokine (IL-4) production in cell culture of Peripheral Blood Mononuclear Cells (PBMC) stimulated by phytohemagglutinin (PHA) derived from 38 patients with chronic C hepatitis and 22 healthy individuals. The percentage of IFN- +cells and IL-4+ cells was measured by flow cytometry. Our results showed that HCV patients group show predominant type 2 response with higher proportion of their CD4+ cell producing IL-4 (18.31 + 5.2%) (mean + SEM) with relative impaired type 1 response with lower proportion of their CD4+ cell producing IFN- (6.36 + 2.32%). Highly significant difference between patients group and control group as regard Th2 response (4.74 + 2.3%) (P> 0.001). Significant difference between patients group and control group as regard Th1 response (2.38 + 0.61%) (P> 0.05). In the present study, there is significant positive correlation between viral RNA titer and Th2 response (r = 0.45; P> 0.05), while a significant negative correlation between viral RNA titer and Th1 response (r = -0.51; P> 0.05) these results indicate that T-helper cells are activated during HCV infection. The predominance of Th2 cell cytokine may be one mechanism where by the host immune response is compromised in chronic HCV disease.

 

 

20/16 STIMULATION UNSATURATED FATTY ACIDS PERMEATED SUPPLEMENTATION TO STATIONARY PHASE BAKER’S YEAST BY LYSINE AND BIOTIN

Sh. Sharaf El-Deen

Microbial Genetics Dept. National Research Centre, Dokki, Cairo, Egypt

The Saccharomyces cerevisiae is used to investigate the reaction of different concentrations of fatty acids supplementations from flax seed oil remnants or from grinding flax seed or with arachidonic acid. The two yeast strains have mutants I some amino acid required e.g., lys1 The fermentable medium contained biotin and selective medium contained 0.1% linolenic acid and 0.4 ml tween 40%. The selected colonies were pitching with flax seed source within stationary phase .The other side of supplementation with arachidonic acid as a one of poly unsaturated fatty acids (AA, 20: 4n-6). The cell viability of two strains affects supplementations fatty acids binding both biotin and lysine .The cell number of LBC strain increased by 50% when 1.0% flax was added but, its increase by 261.5% with 0.5% arachidonic acid in pitching culture. The cell number related with their weights. As well as, their genotypes of strains appropriate a role of supplementations responsibility. The strain LBC has less mutant of amino acid required was high response for supplementation. Free fatty acids fractionation by Thin Layer Chromatography (TLC), and u.v. absorption at 535nm, its were high absorbance with flax supplied 1.5%.But its high absorbance with arachidonic acid supplied 0.7%. The results performed that the fatty acids supplied either flax or arachidonic acid have effects on subsequent fermentation cycle. Free fatty acids content increased by lysine provided (30 mg/l), their absorbance was increased by 60% after 72 h incubation with 1.5% flax. But its did not developed significant response with arachidonic acid The free fatty acids composition determined by GC/MS showed that the colonies without supplementations were contain 2.426% of C18H34O3 and 4.324% of C25H38O2. Its contained 0.202% of C28H44Oand short length fatty acids when flaxsupplied 0.1%. Free fatty acids trends to increase long chain e.g. octadecanoic acid and oleic acid beside the increase of ergosta; the ergsterol of yeast (0.433%). The fatty acids supplementations did not always affect the cells ability to consume glucose. Glucose consumption decreased with flax supplied 1.5%, but its high response with 0.7%arachidonic acid supplied.

 

 

21/16 PMR AND MASS SPECTROSCOPIC DETERMINATION OF ATENOLOL IN PRESENCE OF ITS IMPURITY

M.M.F. Metwally

Analytical Chemistry Department, Faculty of Pharmacy, Zagazig University, Zagazig, Egypt.

Pmr method and mass spectrometry were used to identify and to determine, atenolol and its synethetic impurity percentage. The pmr method involves the measure of peak integeral of atenolol at d = 4.99 of the secondary amine hydrogen and related to the peak integral of maleic anhydride at d = 7.1 in DMSO d6. The % recovery of atenolol was found to be 99.653 + 1.90 (X + S.D.). The mass spectrometry involved identification of the spectrum of the   product resulted from the reaction of N, N-dimethyl-aminobenzaldehyde and atenolol and the percentage of manufacturing impurity was calculated.

 

22/16 construction of mucoadhesive semisolid formula containing chlorhexidine and benzydamine for treatment of periodontitis in patients

S.A. Tayel, M.A. El-Nabarawi, D.A. El-Sotohi, M.A. Ramadan*,

A.M Ezz Al-Arab** and A.S.Gawish***

Department of Pharmaceutics and Industrial Pharmacy, *Department of Microbiology and Immunology, Faculty of Pharmacy, **,***Department of Oral Medicine and Periodontology, Faculty of Oral and Dental Medicine, Cairo University

and El-Azhar University

This study aimed to construct a mucoadhesive semisolid formulation containing chlorhexidine hydrochloride (CHX) and benzydamine hydrochloride (BH) for treatment of inflammatory oral periodontitis in comparison to Corosdyl® gel in patients. Three semisolid formulations were chosen from our previous study, these are 4% hydroxypropyl methylcellulose(HPMC), 5% methylcellulose and glycerin of starch, containing CHX in concentration of 1%w/w. BH is added to these formulations in concentration 0.3%w/w. The viscosity, pH measurement, stability at room temperature for two years, mucoadhesion and the release of both drugs in distilled water were studied. The anti-microbial activity of different formulae containing both drugs was evaluated against different microorganisms. HPMC formula containing both drugs was a perfect base as it gives the higher value for mucoadhesion, release data and anti-microbial activity than Corosdyl® gel. Also, the addition of BH to CHX in the formula does not affect its anti-microbial activity against the tested species. Clinical studies on twenty human patients were done in Periodontology Department, Faculty of Oral and Dental Medicine, Cairo University. The plaque index and gingival index were reduced significantly after treatment. The combination therapy between CHX and BH in HPMC formula is successful for the treatment of periodontitis and prevents its combined inflammations.

 

 

23/16 A PHARMACOECONOMIC STUDY ON AZITHROMYCIN CAPSULES

N.N. Afifi*, M.M. Maher** and A.M. Seleight***

* Department of Pharmaceutics and ** Department of Microbiology, Faculty of Pharmacy, Cairo University

*** Department of ENT, Faculty of Medicine, Zagazig University

The prominent variation in prices of marketed products of the same medicament in relation to its efficiency is of special importance to vital drugs including antibiotics. This is investigated in the present work regarding the broad spectrum azalide antibiotic, azithromycin. Six capsule brands of azithromycin are found in the Egyptian market with wide price diversity. The effect of varying additives on drug release and its anti-bacterial efficiency is studied. Eight adjuvant combinations were involved in preparing azithromycin capsule formulae according to 23 factorial design. The adjuvants used are those found in the different marketed capsule brands, in comparable amounts. Drug released was assayed microbiologically. This allows mutual evaluation of the anti-bacterial activity of the released drug in presence of the used additives. The ultimate step providing anti-bacterial efficiency data was carried out clinically. Prospective and retrospective clinical studies were performed to evaluate the clinical efficiency of the studied products. The applied 23 factorial design revealed that the calculated effects are practically unimportant and most of them are insignificant. This implies that varying additives among different azithromycin capsule formulations does not lead to practically important differences in drug release as long as the official specifications are obeyed. This verifies the absence of any relation between price discrepancies and drug release parameters observed for the marketed formulae which obeyed the official drug release limits. They were concluded to be equivalent regarding drug release and anti-bacterial efficiency. Such in-vitro findings are in accordance with the clinical equivalence of the six marketed capsule products as shown by the prospective clinical study and emphasized by the retrospective one. Accordingly, a cost-minimization analysis is adopted where it has been clearly established by the presented study and provided data that the compared products yield the same health effects. One can then concentrate on costs only in choosing among them.

 

24/16 COMPARATIVE STUDY BETWEEN TRANSDERMAL GLYCERYL TRINITRATE AND BETA-MIMETICS IN THEMANAGEMENT OF PRETERM LABOR

M.F. Midan, M.T. Ismael, K.A. Khattab and M.A. Kolkailah

Department of Obstetrics & Gynecology, Faculty of Medicine, Al-Azhar University

To compare the efficacy and safety of transdermal glyceryl trinitrate and intravenous (I.V) ritodrine as tocolytics in themanagement of preterm labor. This is a prospective randomized study on one hundred (100) cases of preterm labor (PTL), between 28 and 35 weeks' gestation using transdermal glyceryl trinitrate or intravenous ritodrine as tocolytic agent. Treatment was continued till contractions stopped for at least 24 hours. Primary outcome was defined as the time from initiation of therapy to delivery censored at 37 weeks' gestation. Secondary outcome was the proportion of women who delivered after 24 hours between 28-31 weeks, 32-35 weeks, and after 36 weeks’ gestation. There was no significant difference in primary and secondary outcomes after the use of both drugs but most of the maternal adverse events were more frequent and significantly higher in the ritodrine group. Glyceryl trinitrate may be considered a safe and effective tocolytic alternative to ritodrine in the management of preterm labor.

 

25/16 INFLUENCE OF SOME COMMERCIAL OILS ON GROWTH OF FUNGI AND THEIR DEGRADING ENZYMES

S.K. Hemida

Botany Department, Faculty of Science, University of Assiut, Assiut, Egypt

Ten commercial oils (aloe, black cumin, castor, clove, coconut, Eucalyptus, myrrh, olive, paraffin and sesame) were selected to evaluate their effects on fungi. All oils were found to be contaminated with fungi except clove and myrrh oils. Aspergillus flavus, A. fumigatus, A. niger, Penicillium chrysogenum and P. corylophilum were the most prevalent fungi associated with oils. Highest counts of soil fungi were recorded on medium containing 1% castor, olive or sesame oil as a sole carbon source, whereas low counts or no fungal growth at all was observed on clove, myrrh and paraffin oils. The effect of these three oils on lipase and protease productions by some fungi was also investigated.

 

 

26/16 ISOALTION AND IDENTIFICATION OF CRYPTOCOCCUS NEOFORMANS FROM MILK AND PIGEON DROPPINGS, WITH SPECIAL EMPHASIS ON THE THE EFFECT OF OILS ON ITS GROWTH AND CAPSULAR SIZE

R.M. Al-Arosi*, W. Tawakkol**, M.H.R. Kotb* and M.K. Refai***

*Reproductive Diseases Department, Animal Reproduction Research Institute, ** Department of Microbiology, Faculty of Pharmacy, Cairo University, *** Department of Microbiology, Faculty of Veterinary Medicine, Cairo university

Mycological examination of 200 milk and 200 pigeon droppings samples revealed the isolation of Cr. neoformans from 13 milk samples (6.5%) and 20 pigeon droppings (10%). The Canavanine Glycine Bromothymol Blue medium (CGB) was used for the biotyping of the isolates. Twenty-nine out of 33 isolates were biotyped as Cr. neoformans var. neoformans (serotype D or AD) or Cr. neoformans var. grubii (serotype A) and four isolates were biotyped as Cr. neoformans var gattiiand (serotype B or C). The serotyping using reciprocal hyperimmune sera by the slide agglutination test confirmed the 29 isolates as serotype A and 4 as serotype B. The study of the effect of cinnamon, fennel, rosemary, camphor, and thyme oils on the growth and the viability ofCr. neoformans indicated an inhibitory effect for some of these oils (cinnamon, camphor, and fennel), while other oils caused marked increase in capsule size (camphor and rosemary oils). On the basis of these results further studies of these oils are proposed to evaluate their possible therapeutic value. On the other hand, a medium, containing camphor or rosemary oil or both may be used to increase the capsule size of the yeast, which will be helpful in the rapid identification of Cryptococcus neoformans.

 

27/16 STUDY THE EFFECT OF TABLET EXCIPIENTS ON THE PREPARATION OF OTC IBUPROFEN DISPERSIBLE TABLETS USING FRACTIONAL FACTORIAL DESIGN

I.I. Soliman

Department of Pharmaceutics, Faculty of Pharmacy,

Cairo University, Cairo, Egypt.

One half 33 factorial design (15 experiments) was used to study the influence of different excipients on the preparation and the in- vitro evaluation of OTC 100 mg ibuprofen dispersible tablets.The excipients used were: disintegrants, namely, pharmaburst and ammonium carbonate in a concentration of 15% w/w and sodium starch glycolate (primojel) in a concentration of 3% w/w, lubricants, namely, magnesium stearate, propylene glycol and glyceryl behenate (Compritol 888 ATO) in a concentration of 1% w/w and the fillers, Avicel PH 101, maize starch and spray dried lactose in amounts to complete the total weight of the tablets to 200 mg. The prepared OTC ibuprofen dispersible tablets were evaluated by the determination of weight variation, content uniformity, friability, disintegration time, and the fineness of dispersion of tablet as well as the amount of ibuprofen dissoluted during the test of fineness. In addition, infra red spectroscopy was performed to predicte the chemical compatibility between the drug and the excipients used. The obtained results revealed that, the best ibuprofen dispersible tablet formulation regarding the disintegration time (response 1) was obtained when the filler was used at the low level (Avicel PH 101), lubricant at the high level (glyceryl behenate) and disintegrant at the high level (sodium starch glycolate). All tablet formulations containing disintegrant at medium level ( ammonium carbonate) exhibited the highest percentage ibuprofen dissolved in water after 20 minutes (response 2). All tablet formulations containing the filler at low level (Avicel PH 101), lubricant and disintegrant at different levels showed not less than 99% ibuprofen dissolved in Sorensen’s phosphate buffer of pH 7.2 after 20 minutes (response 3).

 


28/16 INTRODUCING A BIOADHESIVE VAGINAL TABLET FORMULATION AS A NEW DOSAGE FORM OF OFLOXACIN

N.N. Afifi* and K.M. El-Houdaiby**

* Department of Pharmaceutics, Faculty of Pharmacy, Cairo University

** Department of Gynecology and Obstetrics, Faculty of Medicine,

Ein-Shams University

Formulation of ofloxacin bioadhesive vaginal tablets is carried out in this work aiming at maintaining a prolonged local drug level at the site of infection thus allowing lower dosing frequency and less amount of administered drug. Moreover, such dosage form provides mainly local treatment of lower genital tract infections in clinical cases where systemic ofloxacin is avoided. It also synergizes systemic treatment in obstinate infections. Different   formulations were prepared by direct compression of 200 mg ofloxacin and 300 mg bioadhesive polymers including sodium carboxymethylcellulose (Na CMC), methylcellulose 1500 (high viscosity MC) polycarbophil (PC), carbopol 934P (C934P) and hydroxypropyl methylcellulose 4000 (HPMC K4M) as well as their blends in different proportions. Stepwise adjustment of such proportions was made according to the results of ofloxacin release study. The prepared formulae were investigated regarding drug release in pH 4.8 McIlvaine's citric acid phosphate buffer using USP dissolution apparatus II. Kinetics of drug release was studied. Formulations of suitable sustained release patterns were evaluated regarding bioadhesion strength as well as bioadhesion time. The formula of choice was tested for bioadhesion time in-vivo. The effect of storage on drug release and its kinetics as well as bioadhesion and tablet hardness was explored for the tablet of choice. Clinical evaluation of the introduced ofloxacin bioadhesive vaginal tablet in comparison to the conventional oral tablet was carried out. Cure rates as well as proportions of recurrent infection were compared and the significance of the difference was determined statistically. Most of the prepared formulae showed zero-order drug release, including the tablets of choice. These contained blends of 65 mg C934P and 235 mg Na CMC, 50 mg HPMC and 250 mg Na CMC, 50 mg PC and 250 mg MC and 50 mg C934P and 250 mg MC. Bioadhesion tests were most satisfactory for the formula containing 50 mg HPMC and 250 mg Na CMC. It retained its properties during storage. The introduced formula showed comparable clinical efficiency to the conventional oral tablet with the advantages of lower dosing frequency, less amount of administered drug and mainly local rather than systemic effect.

 

29/16 PRODUCTION, PURIFICATION AND CHARACTERIZATION OF THE ANTIFUNGAL AGENT OF STREPTOMYCES WERRAENSIS

A.L. Kansoh*; M.R. Abu Shady; F.M. Elbeih; A.A. Elgammal* and A.A. Keera*

*Microbial Chem. Dept. National Research Centre (NRC) and Botany Dept. Fac. Sci. Ain Shams University, Cairo, Egypt.

Streptomyces werraensis was chosen from various identified Streptomyces strains locally isolated from different soil regions of Egypt. It is capable of producing antifungal activities against various moulds and yeasts. S. werraensis showed the highest level of antifungal agent in the culture broth after 4 days of incubation in shake culture. The high level of the antifungal agent production was supported by starch as a carbon source at a concentration of 15g/l., potassium nitrate (2 g/l) as a nitrogen source and dipotassium hydrogen phosphate (1.5 g/l) as a phosphorous source. The antifungal agent was adsorbed on charcoal and silica gel but trials to elute it were unsuccessful. The antifungal agent was best extracted by diethyl ether at neutral pH and was purified on a Sephadex LH-20 chromatographic column. Elementary analysis indicated the absence of nitrogen. The empirical formula was C5H8O and the molecular formula was C30H48O6. The effective purified compound was identified by ultraviolet, infrared, 1H-NMR, 13C-NMR and mass spectra. The spectral characters suggested that this compound was closely related to butyrolactols.

 

 

30/16 IN-VITRO AVAILABILITY AND ANALGESIC ACTIVITY OF MARKETED LOCAL AND IMPORTED PARACETAMOL TABLETS IN COMPARISON TO AN INTRODUCED FORMULA

N.N. Afifi* and A. Mahdy**

* Department of Pharmaceutics, ** Department of Pharmacology,

Faculty of Pharmacy, Cairo University

This work aims at explaining the variability in paracetamol therapeutic performance from different marketed local and imported brands of tabletted dosage forms. The effect of different excipients on the drug in-vitro availability, known as a major cause of such variability, is studied via a 23 factorial design. Avicel PH 101 and corn starch were chosen as fillers. Primogel and Ac-di-sol were used as disintegrants. Magnesium stearate and PEG 6000 were selected as lubricants. Rate and extent of paracetamol dissolution from the eight prepared tablet formulae in modified simulated gastric fluid were used for evaluating the effects and interactions of the studied adjuvants on the in-vitro drug availability. The best qualitative and quantitative excipient combinations could be also introduced as tablet formulations. The in-vitro drug availability from the tablet formula of choice is compared to that from some marketed tabletted dosage forms. Their analgesic efficiency is also compared using writhing test in mice. The results showed that the effect of the tested adjuvants on the rate and extent of paracetamol release from tablets was not significant. However, the calculated effects on the rate of drug release were high. This is in accordance with the observed similarity in the percent of drug released after 2 hours from the tested marketed formulae. They could be discriminated according to the rate of drug release. The imported product X showed the highest rate of drug release followed by the local brands Y then Z. The formulated tablet "c" containing corn starch as filler, Ac-di-sol and PEG 6000 as lubricant and as disintegrant excelled all marketed tested brands. The analgesic efficiency in mice followed the same sequence of drug release rate for products X, Y and Z. The introduced formula "c" showing the highest release rate gave intermediate analgesic efficiency similar to product Y which includes the drug substance from the same source. The in-vitro availability is of only qualitative importance in guaranteeing analgesic activity where all tested formulae exerted significant analgesic effects.The quality of the drug substance, most probably, has the upper hand in determining the therapeutic performance of paracetamol.

 


31/16 CIRCULATING INTRACELLULAR ADHESION MOLECULE-1, VON WILLEBRAND FACTOR (FACTOR V111 RELATED ANTIGEN) AND SKIN HISTOPATHOLOGY AS A DIAGNOSTIC PROSPECT OFRHEUMATOID VASCULITIS

A.M. Dosouki*, M. Shees*, M.I. Sayed*, M.I. Abd-Elkareem**

and G. Abd-Elhameid***

Microbiology and Immunology* Rheumatology, Physical Medicine & Rehabilitation** and Histopathology*** Departments, Faculty of Medicine,

Al-Azhar University

Vasculitis is an inflammatory process that affects the blood vessels leading to damage of the vessel wall. It is one of the most serious events that may accompany or complicate rheumatoid arthritis (RA). The mortality rate among rheumatoid vasculitis (RV) patients is higher than that in RA alone, hence the need for accurate method for early detection of RV is recommended. This study was carried out on 38 patients suffering from RA according to revised criteria of ARA for RA diagnosis. In addition to 15 healthy subjects of age and sex matched with the patient group as controls. The aim of this study was to delineate the role of estimation of the level of von WILLEBRAND   antigen (vW Ag) (Factor V111 Related Antigen) and intracellular adhesion molecule I (ICAM-I) in patients with RA for early detection of RV and to make correlation study between the levels of these two factors with different clinical data and other laboratory studies and skin histopathology. The results of this study revealed presence of clinical vasculitis in only 3 patients (7.9%). The mean   values of vW Ag (F-VIII-R:Ag) and ICAM-I were significantly elevated in RA patients (202 ± 98.4and 448 ± 102 respectively) than the controls (78.5 ± 12.3 and 221 ±6 8 respectively) (P<0.001 for both). The level of vW-Ag (F VIII-R:Ag) and ICAM-I in RA patients with clinical vasculitis were 401 ± 106 and 758±2l8 respectively while in RA without clinical vasculitis were 184.9 ± 92.6 and 421.4 ± I16 respectively, the difference was statistically significant (P<0.05 for both). The differences between the level of vW Ag (F VIII-R: Ag) and ICAM-1 in RA patients with histopathological skin vasculitis and those RA patients without histopathological skin vasculitis were statistically significant (P<0.05 for both). From the result of this study we can conclude that, the estimation of the levels of vW Ag and ICAM-I is suitable methods for early detection of RV.

 

32/16 A COMBINED DRUG THERAPY FOR THE TREATMENT OF HUMAN BACTERIAL INFECTION

N.A. Sabri

FROM

Department of Clinical Pharmacy, Faculty of Pharmacy, Ain Shams University, Cairo, Egypt.

Amoxicillin trihydrate–ibuprofen (A-I) physical mixture at a ratios of 1:1 and 2:1 were subjected to in-vitro examination, for possible drug–drug interactions, using infrared spectroscopy, ultraviolet scanning, differential scanning calorimetry, thin layer chromatography and microbiological examinations. All the previously mentioned examinations confirmed the compatibility of A-I with the absence of any possible interactions. However, the results of microbiological examination showed that ibuprofen potentiate the antibacterial activity of amoxicillin trihydrate towards certain micro-organisms. Thus, sixeteen different capsule formulations containing A-I mixture were prepared, using a fractional factorial design (1/8) of 27 factorial design, and tested for powder content characteristics, amount dissoluted of amoxicillin trihydrate and ibuprofen from the prepared A-I, using first and second derivative spectroscopy and compared with that amount of amoxicillin trihydrate and ibuprofen dissoluted from capsules containing each drug alone. The obtained results showed that neither drug affect the dissolution process of the other. Data treatment and statistical analysis of both powder characteristic and dissolution data was performed, the responses of each run and effect of each variable in the used fractional factorial design was determined, and the significance of each effect was determined using a t-test and variance (S2) of the error was calculated. The chosen formulae of A-I capsules (F4 and F15) were subjected to clinical study on twenty-five patients, pediatrics and adults, suffering from acute tonsillitis and pharyngitis. The patients were divided into two groups, the first group was treated with   the chosen formulae of A-I capsules(combined treatment), the second group was given the prepared amoxicillin trihydrate capsules(single treatment). Data collected before and after treatment include; age, diagnosis, body temperature, total white blood cell count, laboratory data levels for kidney and liver functions. Statistical analysis of the body temperature and the W.B.C. count data by using paired t-test revealed that there was a highly significant difference between the data recorded before and after treatment and between the single treatment and the combined one in decreasing the W.B.C. count to or near the normal level at p< 0.001. Besides, applying the independent t-test it is observed that the combined treatment was much highly effective in combating the infection signified by a higher percentage decrease in the W.B.C. count , mean + S.E. was 45.88+1.11, compared to 53.83+1.27 in case of single treatment . Moreover, it was found that there was no effect of sex on the percent decrease in the W.B.C. count in the single and combined treatments by using independent t-test at p< 0.05. Finally, the recorded values of the liver and kidney functions after A-I capsules administration were all in the normal levels.

33/16 EVALUATION OF THE HYPOTENSIVE EFFECT OFDIFFERENT ATENOLOL HYDROCHLORIDE PREPARATIONS

BY

I.I. Soliman and N.A. Sabri*

Department of Pharmaceutics, Faculty of Pharmacy,Cairo University,. Cairo, Egypt.

*Department of Clinical Pharmacy, Faculty of Pharmacy, Ain Shams University, Cairo, Egypt.

Atenolol HCl sustained release suppositories were prepared using thermally gelling mucoadhesive liquid suppository and double layer suppository. In the former, pluronic F-127 and F-68, representing liquid suppository base, sodium alginate (0-0.9%) as a mucoadhesive polymer and sodium chloride (0-0.9%) as a strength controlling agent were used. The prepared atenolol HCl liquid suppositories were evaluted by testing their gelation temperature, rheological properties and in-vitro release study. In case of atenolol HCl double layer suppositories, an adult suppositories of 2gm containing 100mg atenolol HCl were prepared in which, polyethylene glycol 6000 or witepsol W35 was used as a suppository base. Eudragits RS100 and RL100, pluronic F-127and sodium alginate were used as retarding materials. They were evaluated by testing their weight variation, content uniformity, softening time and in-vitro release study. The best chosen formulae of atenolol HCl sustained release suppository preparations regarding in-vitro release study were subjected to bioavailability and clinical studies. For comparative study, atenolol HCl conventional suppository as well as a commercial Ateno tablet, 100 mg (EIPICO, Egypt) were studied. The obtained results revealed that there was two formulae of atenolol HCl succeeded in the prolongation of drug action over a period of 12 hours. These two formulae were, atenolol HCl mucoadhesive liquid suppository prepared with a base consists of pluronics F-127and F-68in a ratio of 15:20% w/w, in addition to 0.6% w/w of sodium alginate, and 100 mg atenolol HCl double layer suppository prepared using PEG 6000 as a suppository base, and 20% w/w Eudragit RL 100. These two formulae were subjected to a comparative bioavailability and clinical studies on twenty-four males hypertensive patients. The obtained data showed that the conventional suppository and the oral tablet evoked a hypotensive effect after 45 and 60 minutes and continued for 1.75 and 2.25 hours respectively. In addition, their significant acute hypotensive effect and their maximum effect (Emax) were 6.12+0.08 and 7+ 0.25 mmHg respectively.On the other hand, the maximum effect (Emax) for both the double layer suppositories and the liquid suppositories were 10+ 0.17 and 11.5+ 0.13 mmHg with a tmax of 3 and 3.5 hours respectively. The percentage relative bioavailability using the acute hypotensive effect was calculated and it was found to be 111.1, 312.9 and 264.7 for the conventional, the liquid and   the double layer suppositories respectively compared to the conventional tablet Ateno 100mg (EIPICO, Egypt). The mean values of the decrease in the mean arterial blood pressure (MABP) at each interval for each administration were evaluated by ANOVA one way statistical analysis followed by Duncan t-test. It was found that there was a highly significant difference between the prepared sustained release suppositories and the conventional ones at p<0.001.

 

 

34/16 BIOLOGICAL AND CHEMICAL STUDY OF RHAMNUS LYCIOIDES L. LEAVES GROWING IN EGYPT.

S.E. El-Dondity

Department of Pharmacognosy, Faculty of Pharmacy (boys)

Al-Azhar University, Cairo, Egypt.

The L D50 of 70 % alcohol extract of Rhamnus lycioides L. leaves was carried out to determine the safety margin of the leaves. A double-blind trial comparing different concentrations of ointments prepared from 70 % alcohol extracts of Rhamnus lycioides L. leaves with, standard therapy, flumethasone pivalate ointment and a placebo showed that, the extracts of Rhamnus lycioides L. leaves was effective in treatment of induced eczema in mice. A double-blind clinical trial comparing a 2% ointment prepared from 70 % alcohol extracts of Rhamnus lycioides L. leaves with a 0.2 % flumethasone pivalate ointment and a placebo showed that, the 0.2% w/w of flumethasone pivalate ointment was better than 2% w/w Rhamnus lycioides L. leaves ointment but recurrence is larger in flumethasone pivalate ointment than Rhamnus lycioides L. leaves ointment. The results obtained with the extract were statistically comparable to those obtained with the corticoid therapy. Chemical study to isolation and identification of quercetin, and 2 new flavonol glycosides acetate esters viz., {kaempferol-3-O-[2,3,4,-tri-O-acetyl-a-L-rhamnopyranosyl-(1 ® 3) – 2,4,- di-O-acetyl-a-L- rhamnopyranosyl-(1 ® 6) ]-b -D-galactopyranoside and kaempferol-3-O-[ 3,4,-di-O-acetyl-a-L-rhamnopyranosyl-(1 ® 3) – 2,4,- di-O-acetyl-a-L- rhamnopyranosyl-(1 ® 6) ]-b -D-galactopyranoside}. This is also the first report for isolation of quercetin from this species.            

 

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