Research/Original Articles


Determination of Minimum Inhibitory Concentration (MIC) of Hygromycin B in CHO cells

Fatemeh Naddafi, Maryam Tabarzad, Farshad H. Shirazi

Iranian Journal of Pharmaceutical Sciences, Vol. 15 No. 3 (2019), 1 July 2019, Page 1-6
https://doi.org/10.22037/ijps.v15.40472

Chinese hamster ovary (CHO) cells are considered as the most commonly used host for industrial manufacturing of therapeutic proteins. The aim of this study was to evaluate the minimum inhibitory concentration (MIC) of hygromycin B in both CHO-DG44 and CHO-S cells since hygromycin B resistance cassette can be used for future selection of gene expression in CHO cells. The minimum inhibitory concentration was determined by the microdilution method. In this test, CHO cells were subjected to a range of hygromycin B concentrations for a defined period of time. Moreover, CHO cells were scaled up with no addition of fresh culture medium during the growth–culture cycle to investigate their growth. The MIC for hygromycin B in CHO-DG44 cells was 75 μg/ml and for CHO-S cells was 50 μg/ml. Thus, it has been indicated that CHO DG44 cell line could be more effective for stable cell line development than CHO-S cell line.

Several 4-substituted-benzylidene-6-substituted-phenyl-4,5-dihydro-pyridazin-3(2H)-one derivatives (3a-q) were synthesized and evaluated for their antimicrobial activities with an aim to obtain promising antitubercular agents. In the first step, 6-aryl-2, 3, 4, 5-tetrahydro-pyridazin-3-ones (2) was prepared by reacting 4-aryl-4-oxobutanoic acids (1) with hydrazine hydrate. Then, aryl-aldehydes were reacted with compounds 2 to furnish pyridazinones (3a-q). Finally, the synthesized compounds were evaluated for their in vitro antitubercular activities against mycobacterium strains and compared to reference drugs streptomycin (MIC value of 6.25μg/mL) and pyrizinamide (MIC value of 3.125μg/mL). Compound 3m and 3n was found to have most significant action. Identity of these compounds was ascertained using IR, NMR and mass spectral data results.

Fungus Candida albicans has received much attention due to its oral, vaginal and/or systemic candidiasis.This study was undertaken to find out the optimum concentration of two essential oils of Thymus daenensis Celak. and Zataria multiflora Boiss that have been effective on the C. albicans. The essential oils (EOs) were obtained by hydrodistillation method from T. daenensis and Z. multifera and Time-dependent killing potential of essential oils were assessed in phosphate buffered saline solution containing the desired concentrations of the test agents and yeast suspension. Time of action, concentration of individual or EO mixture and T. daenensis:Z.multifera mass ratio predicted using response surface methodology (RSM) and artificial neural network (ANN).RSM and ANN techniques predicted the 0.8% as an optimum percentage concentration of EO mixture in oils ratio T. daenensis:Z. multifera 1:1, ensuring the highest antifungal effect of 95.8% and 96.4% after 20 h.Appraisal of the models through the coefficient of determination (R2) and mean-square error (MSE) show that the ANN was superior (R2= 0.994) to the RSM model (R2= 0.957) in predicting the percentage of reduced cells.Our data confirmed that proper EOs mixtures may reduce the minimum effective dose of individual EOs.

New N2O2 type Schiff base has been designed and synthesized by condensing 2, 4 dihydroxy benzaldehyde and α-naphthylamine in ethanol. Solid metal complexes of the schiff base with Cu(II), Ni(II) and Zn(II) metal ion were synthesized and characterized by elemental analysis, magnetic susceptibility, molar conductance, IR, UV-Vis and ‘H NMR spectral studies. The data shows that the complexes have the composition of ML type.The UV-Visible and magnetic susceptibility data of the complexes suggest a square-planar geometry around the central metal ion except Zn (II) complex which has tetrahedral geometry. The redox behaviour of the complexes was studied by Cyclic Voltammetry. The in vitro activities of the compounds were tested against Staphylococcus, Bacillus subtilis, Escherichia Coli and Pseudomonas by well-diffusion method and antifungal of the compound were tested against fungi such as Aspergillus niger, Aspergillus flavus and Rhizoctonia bataicola by minimum inhibitory concentration. All the metal complexes showed stronger antifungal and antimicrobial activities than the free ligand.

Profile of Prescribing Practice at A Referral Hospital in Indonesia Using Who Drug Use Indicators

Milati Dyah Utami, Susi Ari Kristina, Abdul Karim Zulkarnain

Iranian Journal of Pharmaceutical Sciences, Vol. 15 No. 3 (2019), 1 July 2019, Page 41-46
https://doi.org/10.22037/ijps.v15.40476

To promote rational drug use in developing countries, assessment of drug use patterns using the WHO drug use indicators is becoming standard in documenting rational prescribing. The aim of this study was to assess the drug prescription patterns at the Outpatient Pharmacy Department of Sleman District Hospital, using the WHO drug use core indicators. A descriptive cross-sectional survey was conducted. The sample was selected using systematic random sampling of 700 patient encounters were reviewed retrospectively for a one year period from January 2017 to December 2017. Data were collected using an investigator-administered data collection form.The main outcome measured is patterns of drug use measured using WHO core drug use indicators. The data obtained showed that, out of total drugs prescribed (2215), the average number of drugs prescribed per encounter or mean was 3.16 (SD 0.89) with a range between 1 and 4. The percentage of encounters in which an antibiotic prescribed was 15.75%. The Percentage of drugs prescribed by generic name and from a national drug list was 95.66% and 97.78, respectively. The most commonly prescribed forms of antibiotics were amoxicillin 10.74%. The number of drug prescribed shows deviation from the standard recommended by WHO. Drug use evaluation should be done for unnecessary medication. Developing guidelines in place for the optimal and responsible use of antibiotics which could be monitored and feedback provided on a regular basis to prescribers.

Evaluation of Analgesic Activity of Ficus Palmata

Skaik Aminabee, A. Lakshmana Rao, K. Sowmya, D. Nymisha, K. Kusuma Naga Lakshmi, K.V.N.S. Manikanta, P. Praveen Kumar

Iranian Journal of Pharmaceutical Sciences, Vol. 15 No. 3 (2019), 1 July 2019, Page 47-60
https://doi.org/10.22037/ijps.v15.40492

Ficus palmata (FI) is an important and widely used medicinal plant. It is principally used as an item of diet in the treatment of constipation and diseases of the lungs and bladder. The sap is used in the treatment of warts. Ficus palmata plant is used in various disease e.g. gastrointestinal, hypoglycemic, antitumor, anti-ulcer, anti-diabetic, lipid lowering and antifungal activities. This study evaluates both the central and peripheral analgesic effect of the different extracts of Ficus palmata in the experimental animals. Methods: Acute toxicity test was done following the Organization of Economic Cooperation and Development guidelines. Ficus palmata extracts (250 mg/kg, 500 mg/kg) body weight was evaluated for central analgesic activity by the hot plate method, tail immersion method and formalin test models using tramodol
(20 mg/kg b.w.) as the standard drug. Results: In all the models, chloroform extract showed significant inhibition as well as the elongation of time at a dose of 500 mg/kg body weight. A linear dose response relationship was also observed which was comparable with that of the standard drug tramodol (p<0.01, p<0.05). Conclusion: The study showed significant central and peripheral analgesic activity of Ficus palmata which may be attributed to the inhibition of prostaglandin synthesis, phospholipase A2, and tumor necrosis factor alpha. Ficus plamata as a commercial source of analgesic drug should be subjected to further research.

Amoxicillin (α-amino-p-hydroxybenzyl-penicillin) is a semi-synthetic, orally absorbed and widely prescribed β-lactam antibiotic. It is now widely used for eradication of gastric Helicobacter pylori infection
combined with a second antibiotic and an acid‐suppressing agent despite its short elimination half-life of one hour. The purpose of this study was to develop and evaluate amoxicillin loaded carbopol 934P mucoadhesive microcapsules for sustained drug release at the gastric mucosa to prolong the residence time of dosage form in the stomach and to achieve controlled drug release for more effective H. pylori eradication. Amoxicillin mucoadhesive microcapsules were formulated by ion gelation technique using 32 factorial designs. A 32 full factorial design was used to derive the statistical equation, ANOVA analysis, contour plots and 3D response surface plots. FT- IR (4000 cm-1- 450 cm-1) analysis of amoxicillin and with polymers was performed by using potassium bromide pellet method. Different polymer ratios of carbopol 934P and sodium alginate were used to formulate by nine formulations (F1 to F9) amoxicillin mucoadhesive microcapsules. The prepared formulations were characterized by determining their percentage of yield, particle size, percentage of entrapment efficiency,swelling index, percentage of mucoadhesion and percentage of drug release. Amoxicillin and used polymers are found to be compatible with no interaction reported by FTIR analysis. The optimized formulation (F9) exhibited a high drug entrapment efficiency of 96.04±0.03%, particle size of 847.2518±0.06 μm, yield of 98.86±0.01%, swelling index of 120.09±0.06%, and mucoadhesion of 67.00±0.02 after 8 h. A successful sustained drug release was achieved for more than 14 h. In-vitro dissolution test for optimized formulation (F9) demonstrated a
slower release behavior in 0.1N hydrochloric acid followed by linear release profile in pH 7.4 phosphate buffer. The drug-to-polymer-to-polymer ratio had a more significant effect on the dependent variables. The release kinetic study of optimized formulation (F9) displayed a good fitting with zero order behavior and Korsmeyer-Peppas model has confirmed a non-Fickian release. The developed mucoadhesive amoxicillin microcapsules were observed to have adhered strongly with gastric mucosa with approximately 14 h of prolonged stay expecting improved bioavailability and reduced dosing frequency and subsequently improving patient’s compliance. The concentration of carbopol 934P and sodium alginate had highly significant effects on depended variables. The present study concludes that amoxicillin mucoadhesive microcapsules can be effectively used for the more effective treatment of H. pylori infection.

Abdominal adhesions are considered as an important problem in abdominal surgery. In this study we compared the effectiveness of atorvastatin, hyaluronic acid and interceed (oxidized cellulose) in prevention of postoperative intestinal adhesions. A total of 40 Wistar-albino rats were randomly divided into 5 groups: Group 1 (Sham); Group 2 (Control or Adhesion induction); Group 3 (Adhesion induction + hyaluronic acid); Group 4 (Adhesion induction + oxidized cellulose); and Group 5 (Adhesion induction + atorvastatin). A same experimental method of cecal abrasion and ligature was used to produce adhesions in all rats. Fourteen days after adhesions formation, animals were killed and adhesions were scored according to macromorphological characteristics and adhesion-carrying tissues under standard histologic examination. Inflammation,vascularization and fibrosis in granulation sites were graded in all samples. Adhesion scores in all experimental groups were slightly lower than control group. Group 3 had the lowest mean adhesions score (1.18 ± 1.24) based on surgeon observation. The mean of adhesion score in Group 3 (1.18 ± 1.24) was significantly lower than that in Group 4 (3.43 ± 0.53) (p=0.004). Group 5 showed the lowest mean adhesions score on pathological examination (5.62 ± 3.73). The mean of adhesion total score in Group 3 was slightly lower than that in other groups; however, this difference was not significant. Hyaluronic acid, oxidized cellulose, and atorvastatin are effective in treatment and prevention of intestinal adhesions during open abdominal surgery. Although hyaluronic acid was slightly more effective, further studies are needed to evaluate the effect of combination therapy with these agents.