Vol. 2, June, 1997.

1/2 QUINAZOLINE ALKALOIDS FROM

ANISOTES TRISULCUS L.

M.M. Al-Azizi

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

Nasr City, Cairo, Egypt.

The quinazoline alkaloids, anisotine 7, vasicinone 2 and peganine 3_ were isolated for the first time, from the aerial parts of A. trisulcus L. The chemical strucutre of the isolated alkaloids were characterized by ir, uv, pmr, andcmr as well as their mass spectral data. C-NMR of anisotine 7 is reported for the first time. A revision in some of the 'H-NMR spectral assignments of anisotine / and peganine 2 •were also made.

2/2 A SPECIES-SPECIFIC DNA PROBE FOR PROTEUS MIRABILIS IDENTIFICATION

M. S. E. Ashour, M. I. Khan* and Moselhy S. Mansy

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

Nasr City,Cairo, Egypt and *Department of Pathobiology, College of Agriculture and Natural Resources, University of Connecticut, Storrs, CT 06269-3089, USA.

Endonuclease Hind III fragments of Proteus mirabilis chromosomal DNA was cloned in Escherichia coli using the plasmidpUC 18. A 3. 5-kilobase-pair (kbp) DNA fragment specific for Proteus mirabilis was identified by colony, dot, and Southern hybridization analyses. When labeled and used as a probe, this recombinant clone hybridized with the DNA of 18 Proteus mirabilis, but did not hybridize with the DNA of 71 strains of Gram-negative and Gram-positive bacterial species. In Southern blot hybridization, this probe hybridized specifically to a 3. 5-kbp Rind Ill-digested DNA fragment of 18 P. mirabilis isolates from human as well as animal sources.

3/2 POLYMERASE CHAIN REACTION-BASED RESTRICTION

FRAGMENT LENGTH POLYMORPHISMS (PCR-BASED

RFLPS) TYPING OF TRIBE PROTEEAE SPECIES

M.S.E. Ashour, M.I. Khan* and M.S. Mansy

Department of Microbiology, Faculty of Pharmacy, University of Al-Azhar, Nasr

City, Cairo, Egypt and *Department of Pathobiology, College of Agriculture and Natural Resources, University of Connecticut, Storrs, CT 06269-3089, USA.

Fifty-nine strains representing the ten reco~iized tribe Proteeae species were examined by restriction fragment length polymorphisms (RFLPs) analysis of an 1.5-kbp PCR-amplified portion of the 16Sribosomal RNA gene in tribe Proteeae. Of 13 restriction endonucleases, digestion of purified PCR products with Msp 1 and then digestion with Hha I produced RFLPs band patterns which separated all tribe Proteeae species and to differentiate between several strains of the tribe Proteeae. DNA amplification with RFLPs analysis is the first rapid method that distinguishes all species and recognized strains within the tribe Proteeae.


4/2 SAFETY OF IMMUNIZATION BY rDNA HB VACCINE IN EGYPTIAN HEALTHY ADULTS

M.S.E. Ashour, K.S.F. Abd El-Wahab,* M.M. Ashour **

and S.F.M. EI-Kastawy

Microbiology Department, Faculty of pharmacy, * Microbiology Department,

Faculty of medicine (Girls), Al-Azhar university. ** Tropical Medicine Department,

Faculty of Medicine, Ain Shams University

The availability of safe and effective hepatitis B vaccines allows the establishment of immunization programme on a global scale, leads to the elimination of hepatitis B and the reduction of mortality due to its sequelae. The results revealed that very mild adverse reactions were recorded after the first dose only and were tolerated within two days. No side effects were observed in any individual after the second and third dose of the vaccine. Regarding the effect of hepatitis B vaccine on blood picture and liver functions, there was insignificant difference between vaccinated and control groups either at the start or at the end of the study, and also after each dose of the vaccine. Hepatitis B vaccine had therefore no effect on blood picture and liver functions of the vaccinated group. Therefore the HB vaccine is safe and effective and no side effect was recorded after vaccination.


5/2 ACCELERATED STABILITY TESTING AND SHELF-STORAGE OF NITROFURANTOIN SOLID DISPERSION IN TABLET AND CAPSULE FORMULATIONS.

H.H. El-Shattawy, A.A. Kassem, A.M.S. Ahmed and A.M. El-Gendy Faculty of Pharmacy, Al-Azhar University, Nasr City, Cairo Egypt.

Accelerated stability testing (AST) at 35 and 45°C as well as shelf-storage at room temperature for eleven months -were performed on six Nitrofurantoin tablet and capsule formulae that met the dissolution tolerances of the USP XXIII and having reasonable dissolution patterns in addition to the commercial Nitrofurantoin tablets (Colifuran). Shelf-storage stability testing obviates that all the investigated tablet and capsule formulae met the pharmacopoeial requirements for drug content after storage for 11 months. The percentage drug remained were 95.84-98.06 for the prepared formulae and 98.96 for the commercial tablets. The AST could be of value in rating the stability of the tested Nitrofurantoin formulae but one cannot rely on in calculating their expiry. Formula IV (Tablets containing citric acid-carrier--Avicel PH 101 in a ratio of 1: 0.5:1.5 with 4% talc, 1.2% stearic acid and 2. 4%Aerosil200 was found to be the best base for Nitrofurantoin tablets concerning the dissolution data, shelf-storage stability, percent drug excreted and relative bioavailability.

6/2 T-LYMPHOCYTES AND ITS SUBSETS IN ACUTE RHEUMATIC FEVER AND ACTIVE RHEUMATOID ARTHRITIS

M.M.M. El-Sadawy*, Saad Ibrahiem Def Alia**

and Mohei-Eldean H. Ahmed***

Microbiology*, Cardjology*.* and Rheumatology and Physical Medicine*** Departments, Faculty of Medicine, Al-Azhar University, Cairo, Egypt.

Thirty patients suffer mg from acute rheumatic fever (RF), 30 active rheumatoid arthritis (RA) and 30 apparently healthy age and sex matched controls were the subject of this study. Peripheral blood T-lymphocytes and its subset, were enumerated using monoclonal antibodies (M-A b) specific against 1-cells, T-helper/inducer and Tsuppressor/cytotoxic, (T3, 14 & T8) and indirect immunojluorescent, technique, in ARF patients a statistically significant (P<0.05) leucocytosis was observed when compared to controls. However, significant decrease ofT3 (P<0.05), T4 (P<0.05) and T8 (P<0.05) were also illustrated. On the other hand insignificant decrease of total lymphocytes and 74/18 ratio were detected. Among ARA patients, statistically significant increase ofT4 (P 0.05) and T47T8 (P 0.05) ratios were observed when compared to healthy subjects. On the other hand significant leucopenia (P<0.05) and significant reduction ofT8 cells (P< 0.05) were also noticed However the number of total lymphocytes and T-cells (73) showed insignificant reduction (P>0.05) when compared to controls.

7/2 HEPATITIS C VIRUS GENOTYPES IN SOME EGYPTIAN PATIENTS USING REVERSE HYBRIDIZATION ASSAY

M.A. Saber, I. El-Dabaa, M. Abdel Hady* and M.H. Romeih

Biochemistry Department, Theodor Bilharz Research Institute and Microbiology

Department*, Faculty of Medicine, AI-Azhar University

Because of the enormous variability of hepatitis C virus ('1CV); the different profiles of pathogenicity; infectivity and response to antiviral therapy, the development of reliable genotyping assays is a formidable challenge. The optimal genotyping region appears to be the 5" untranslated region (5' UTR) because of high conservation within, but considerable variability between, genotypes. To investigate the genotypes and subtypes ofHCVin Egypt, sera from 40 patients with chronic HCV infection were investigated; 36 males and 4 females. Their ages ranged from 20 to 67 years. All patients were seropositive for anti-HCV by ELISA and positive for HCV-RNA by reverse transcription nestedpolymerase chain reaction (RT-PCR). The HCV genotypes and its subtypes were determined by reverse hybridization Inno-LiPA (Line Probe Assay). HCV genotype 4 was found to be the most prevalent and represents 95% among serum samples analysed. Its subtypes c/d, c, h and c/d/fwere prevalent in 70%, 5% and 2.5%, respectively, while genotype 4 without definite subtype was found in 12.5%. In addition, HCV genotypes and subtypes Ib and 2b were equally detected in 2.5% of patients. In conclusion, HCV genotype 4 especially subtypes c/d is the most prevalent in Egypt during the year 1996 within the studied group.

8/2 EFFECTS OF METAL IONS AND PH ON THE ACTIVITY OF

FLUOROQUINOLONES AS ERADICATING AGENT AGAINST

BIOFILM ASSOCIATED PSEUDOMONAS AERUGINOSA

M.A.M. Yassien

Department of Microbiology and immunology, Faculty of Pharmacy, Ain Shams University, Cairo, Egypt.

The   effect of metal ions   (Ca++   and Mg++) and pH on the activity of norfloxacin,   ciprofloxacin,   pefloxacin and ofloxacin, as eradicating agents, was studied against preformed bio films of Pseudomonas aeruginosa in wells of microtiter plates and on segments of urinary catheters in an in vitro model of catheter colonization.   The addition of fluoroquinolones (6.25-100 jug/ml) alone, with 5 mM Ca++, and with 5mM Mg++ to the preformed biofilms of P. aeruginosa in the micro titer plates caused reduction in the optical density of the bioflims to 44-84 %, 47-88%, and 64-95% of the control, respectively. The experiments were repeated using fuoroquinolone solution adjusted to different pH. 1 'reatment of the adherent biofilms with norfloxacin and ciprofloxacin adjusted at pi 1 5.5, 7, and 8.5, caused reduction in the optical density to 53-79%, 61-88%, and 68-93% of the control (untreated biofilms), respectively. In the in vitro model of catheter colonization, the same effect of metal ions and pH was observed These data showed that the presence of 5 mM Mg++   decrease   the   activity of norfloxacin,   ciprofloxacin,   and pefloxacin as eradicating agent against biofllm associated P. aeruginosa. The inhibitory effect of Mg++ may be either due to reduction in the rate of penetration offluoroquinolones across the biofilm matrix or inhibition of chelation of the Mg++ ions that incorporated in the bio film arid has a role in its stability. Also, rising up thepHof norfloxacin and ciprofloxacin solutions caused slight reduction in their activity as eradicating agent, while, decreasing the pH to 5.5 slightly increased their activity. The effect ofpH is mainly through changing the molecular charge of fluoroquinolones molecules that may have an effect on their penetration through the biofllm matrix.

9/2 PROPOSED MODE OF ACTION OF OMADINE

M.S.E. Ashour, Robert T. Vinopal*, Robert W. Coughlin** and M.A.B. Gamal.

Department of Microbiology, Faculty of Pharmacy, Al-Azhr University Cairo, Egypt

Department of Chemical Engineering and Biotechnology Center,** and Department

of Molecular and cell biology University of Connecticut, Storrs, CT., USA.~

In this study experiments were pe, formed to determine the mode of action of Omadine. Omadine was very effective in inhibiting the growth of all bacterial and yeast isolates examined and the minimum inhibitory concentration values against the tested isolates indicated that in general yeasts and Gram positive bacteria were sensitive to Omadine as demonstrated by their low minimum inhibitory concentration (MIC) values, while Gram negative bacteria tend to be more resistant displaying higher MIC values. Also, Omadine was very effective in inhibiting the growth of cell wall deficient microorganisms (Mycoplasma spp.) and this indicates that the mode of action of Omadine may not be through the action on cell wall. Electron microscopical examination ofE. coll and Bacillus subtilis under action of Omadine revealed that no apparent effect of Omadine on the structure and assembly of the cell wall of each organism, 'as compared with controls. Testing the effect of metal cation on activity of Omadine was done using different concentrations of ferrous sulfate and no apparent effect was detected on the activity of Omadine. Four sets of experiments were performed to determine the ability of Omadine in making the cell membrane leaky. Omadine has no effect on leakage ofpurines, pyrimidines and inorgainc phosphorous, but greatly affect leakage of potassium and magnesium ions. However, Omadine induces leakage of thiomethylgalactoside (TMG) and its activity in causing leakage increases at low pH. Omadine is very active hi reducing cellular adenosine triphosphate levels, and marked 'reduction hi A TP level was clear at low pH values. The effect of increasing the ionic strength of the medium on Omadine activity was studied and only high concentration of potassium ions have to some extent protective effect against the inhibitory action of Omadine. So the proposed mode of action of Omadine acts by catalyzing the electroneutral exchange oflf and other ions with K+ across cell membranes, resulting in collapse ofH* gradients, tC gradients and other ion gradients important to cell function, with consequences depending on the condition and organism. This action was explained by predicting that omadine known to form stable chelation complexes with heavy metal ions, also forms quasi-stable coordination complexes with K* ions.

Scroll to top