Asian Pacific Journal of Tropical Biomedicine

192-197

Authors: Sirada Pongmuangmul, Supaporn Phumiamorn, Phanchana Sanguansermsri, Nalin Wongkattiya, Ian Hamilton Fraser, Donruedee Sanguansermsri

Objective: To evaluate the monogalactosyl diglyceride (MGDG) and digalactosyl diglyceride (DGDG) from Clinacanthus nutans (C. nutans) for their in vitro antiviral activities against herpes simplex virus type 1 (HSV-1) and type 2 (HSV-2) by plaque reduction assay. Methods: MGDG and DGDG were extracted with chloroform from C. nutans leaves. MGDG and DGDG were separated from chloroform crude extract using column chromatography, characterized by thin layer chromatography and quantified by high performance liquid chromatography. The anti HSV-1 and 2 activity against pre-treatment and posttreatment of the compounds was evaluated using plaque reduction assay. The cytotoxicity of the extract and the compounds on Vero cells were performed by MTT assay. Results: MGDG and DGDG obtained by column chromatography showed identical profiles as standard MGDG and standard DGDG using thin layer chromatography and high performance liquid chromatography. MGDG and DGDG from C. nutans showed 100% inhibition of HSV-1 replication at the post step of infection at noncytotoxic concentration with IC50 values of 36.00 and 40.00 mg/mL, and HSV-2 at 41.00 and 43.20 mg/mL, respectively. Moreover,MGDG and DGDG fromC. nutans were demonstrated to have antiherpes simplex activity at the same level as standard synthetic compounds. In contrast, pretreatment of Vero cells with MGDG and DGDG before HSV-1 and HSV-2 infection did not show inhibitory effect against these viruses. MGDG and DGDG exhibited antiviral activity against HSV-1 with selectivity index of 26.00 and 23.00 and HSV-2 of 23.30 and 21.30. Conclusions: MGDG and DGDG from C. nutans, a traditional Thai herbal medicine illustrated inhibitory activity against HSV-1 and HSV-2, probably by inhibiting the late stage of multiplication, suggesting their promising use as anti-HSV agents.

202-210

Authors: Romero Marcos Pedrosa Brandão-Costa, Vivianne Ferreira Ara´ujo, Elizabeth Neves, Maria Tereza Santos Correia, Ana L´ucia Figueiredo Porto, Maria das Graças Carneiro-da-Cunha

Objective: To explore the effects of aqueous extract from Phthirusa pyrifolia leaves (67 mg/kg body weight for 12 days) on the reproductive function of male Wistar rats through oral administration. Methods: Animals (n = 30), aged 13 weeks and weighing (378.5 ± 5.0) g, were housed in a vivarium under controlled environmental conditions [photoperiod of 12 h light/dark, temperature of (23 ± 1)
C] and were fed standard rations ad libitum. The experiment ran for 12 days, wherein animals were divided into three groups: negative control (n = 6) received water, positive control (n = 12) with finasteride at a concentration of 1.0 mg/kg; and a test group (n = 12) submitted to aqueous extract. At the end of the experiment, the animals were sacrificed and submitted to analyses. Results: The morphological results of the testes showed that the aqueous extract induced significant changes in the diameter and cross-sectional area of the seminiferous tubules as well as the thickness of the seminiferous epithelium. Furthermore, the extract was able to abruptly decrease testosterone concentrations by about 81.88% in the treated group when compared with the negative control, (47.0 ± 4.8) ng/dL and (255.0 ± 2.0) ng/dL, respectively, and 76.8%, (211.0 ± 8.7) ng/dL, when compared with finasteride. However, the extract causes neither liver damage nor impairment of renal function. Conclusions: These results suggest that the high amounts of flavonoids shown to be in the extract may be responsible for its hepato-protective effects and suggest a possible decrease in the libido and reproduction of rats.

216-220

Authors: Aussavashai Shuayprom, Donruedee Sanguansermsri, Phanchana Sanguansermsri, Ian Hamilton Fraser, Nalin Wongkattiya

Objective: To establish a simple, rapid, precise and accurate high performance thin layer chromatography (HPTLC) method with densitometric detection for the determination of vitexin in Passiflora foetida Linn. (P. foetida). Methods: Ethanolic extract of the plant leaf powder was used for the experimental work. Separation was performed on silica gel 60 F254 HPTLC plates with ethyl acetate: methanol: distilled water: formic acid in the proportion of 50:2:3:6 (v/v), as the mobile phase. The determination was carried out using the densitometric absorbance mode at 340 nm. Results: Vitexin response was linear over the range of 2.5–17.5 mg/mL with a correlation coefficient of 0.996. Intraday and interday precision studies showed the relative SD was < 3%. Accuracy of the method was determined and the average recovery was 100.3%. The limit of quantitation and limit of detection were 0.879 and 0.290 mg/mL, respectively. The contents of vitexin in P. foetida leaf extracts were within the range of 0.030%– 0.310%. Conclusions: The method was evaluated for sensitivity, accuracy, precision and reproducibility. Each analysis by HPTLC is less expensive than current methods. This method is suitable for routine quality control of raw material of the leaves of P. foetida extract and its products.

225-231

Authors: Ovuakporie-Uvo Oghale, MacDonald Idu

Objective: To study the phytochemistry, anti-asthmatic and antioxidant activities of the aqueous leaf extract of Anchomanes difformis (Blume) Engl. (A. difformis) and to verify claims of use in folk medicine. Methods: For anti-asthmatic activity, male and female guinea pigs with average body weight of (451.4 ± 118.1) g were divided into six groups of six animals each. Group 1 served as control (distilled water); Group 2 was administered with salbutamol (reference drug) only; Group 3 served as ovalbumin sensitized group, Group 4, 5 and 6 were treated with A. difformis extract at doses of 100, 200 and 400 mg/kg, respectively. Described methods were used to test fluid viscosity, fluid volume and quantitative phytochemistry analysis. Absorbance was read using a UV–Vis spectrophotometer and results computed in percentage. Total antioxidant assays [2, 2-diphenyl-1-picrylhydrazyl (DPPH) and lipid peroxidation assay], were carried out using reported procedures. Results: The anti-asthmatic evaluation showed that protection from asthma of the animals in Group 6 (400 mg/kg, 32.7%) were similar to that in Group 2 (salbutamol, 33.0%). Excised trachea was free of mucus secretion in Group 5 (200 mg/kg) as was observed in the control group. Fluid volume increase in Groups 3 and 6 indicated mucus secretion. DPPH radical scavenging activity of extract was effective as ascorbic acid which served as standard at 20 mg/mL. But, the extract elicited low lipid peroxidation activity compared with the reference (tocopherol) at concentrations tested. Conclusions: A. difformis aqueous leaf extract is safe and possesses positive antiasthmatic and antioxidant activities as claimed by traditional herbal practitioners in Delta State.

239-245

Authors: Souhila Mahmoudi, Mustapha Khali, Abderahim Benkhaled, Karima Benamirouche, Imen Baiti

Objective: To determine the total phenolic and flavonoid contents, antioxidant and antimicrobial activities of methanolic leaf extracts of ten Algerian fig (Ficus carica L.) varieties (uniferous, biferous and caprifig tree). Methods: Phenolics were extracted by Soxhlet method and analyzed by the Folin– Ciocalteu colorimetric method. Flavonoids were determined by aluminum trichloride assay and the antioxidant capacity was determined by the 2,2-diphenyl-1-picrylhydrazyl radical scavenging assay. The antimicrobial activity was studied with the disc diffusion method and a macrodilution broth method was used to determine the minimal inhibitory concentrations and minimal lethal concentrations. Results: The mean extract yield was 14.10% ± 0.66% (n = 10). Leaf extract of biferous followed by uniferous varieties had the highest total phenolic contents [(52.296 ± 5.232) and (48.973 ± 2.015) mg gallic acid equivalent/g of dry plant extract respectively], flavonoids [(14.388 ± 0.333) and (14.136 ± 1.082) mg quercetin equivalent/g of dry plant extract] and antioxidant capacity [IC50 (798.754 ± 108.590) and (825.004 ± 110.835) mg/ mL]. Antioxidant capacity of fig leaves was significantly correlated with phenolic contents (r = 0.748). These extracts showed bactericidal activity and moderate antifungal activity, and the minimal inhibitory concentrations and minimal lethal concentrations were determined on Bacillus cereus and Staphylococcus aureus. Conclusions: All tested extracts contain phenolic compounds and exhibited an antioxidant activity and an antimicrobial effect against Gram-positive and Gram-negative bacteria. Further researches on identification and purification of phenolic compounds are required.

251-255

Authors: Agustina Setiawati, Handika Immanuel, Mery Tri Utami

Objective: To determine the inhibition activity of Typhonium flagelliforme Lodd. Blume (T. flagelliforme) leaf extract on cyclooxygenase 2 (COX-2) expression of colon cancer cells. Methods: T. flagelliforme leaf extract was prepared to macerate in ethyl acetate. In vitro anticancer activity was assayed by MTT method on WiDr colon cancer cells. This study applied apoptosis induction assay to investigate the mechanism of cell death using double staining method. COX-2 expression was stained by immunocytochemistry. Results: T. flagelliforme showed anticancer activity and induced apoptosis on WiDr cells through inhibition of COX-2 expression with IC50 70 mg/mL. Conclusions: This study showed that T. flagelliforme is a promising chemopreventive agent for colon cancer through COX-2 inhibition.

259-264

Authors: Jafar Jalaei1, Mehdi Fazeli, Hamid Rajaian, Somayeh Layeghi Ghalehsoukhteh, Alireza Dehghani, Dominic Winter

Objective: To investigate the antihistamine-releasing effect of a peptide isolated from wasp venom of Vespa orientalis. Methods: This peptide was separated from crude venom by chromatography methods and mass spectrometry. Then various concentrations (2, 4, 8, 16, 32, 64, 128 and 256 mmol/L) of the peptide were incubated with mast cells and lactate dehydrogenase assay was performed. Results: No significant effect was observed in lactate dehydrogenase absorbance under 128 mmol/L concentration. This implied that the peptide did not cause cell death in mast cells and consequently, histamine release did not happen. Moreover, the results showed the IC50 of mast cells degranulation at 126 mmol/L, which was approximately high implying that this peptide had high selectivity for normal cells and did not cause histamine release from these cells. Conclusions: This would be a great aim in new drug development, in which an agent acts potentially on its target tissue without activating the immune system.

270-274

Authors: Reham F. El-Kased

Objective: To evaluate the antibacterial activity of Egyptian honey against bacteria causing respiratory tract infections. Methods: Sputum and throat swab specimens were used, from which five bacterial species were isolated, namely, Klebsiella pneumonia, Staphylococcus aureus, Streptococcus pyogenes, Pseudomonas aeruginosa and Streptococcus pneumonia were isolated, identified and grown on suitable media for further identification or confirmation. Different concentrations (100%, 75% and 25%) of honey and simulated honey solution were used for activity assay and estimation of minimum inhibitory concentration and minimum bactericidal concentration. Results: All the tested bacterial isolates were completely susceptible to the 75% concentrations of honey and to the 100% concentration of the simulated honey solution. This may be due to the high osmotic pressure exerted by the high sugar content in both honey samples. Moderate susceptibility of the isolated bacteria to honey at 100% v/v concentration, and resistance to honey at 25% concentration and the 75% and 25% concentrations of simulated honey solution, indicated the presence of other antimicrobial components responsible for the activity other than the osmotic pressure. Therefore, it was suggested that honey showed distinguished antibacterial activities against the most common bacteria causing respiratory infections with varied sensitivity. Conclusions: Honey, a non-toxic, nutritious, safe for human consumption and cheap natural antibacterial agent, should be globalized.

100-107

Authors: Sillma Rampadarath, Daneshwar Puchooa

Objective: To explore the phytochemical and antimicrobial activities as well as the insecticidal properties of the different sections of Ricinus communis (castor) plant in Mauritius. Methods: Qualitative and quantitative methods were used for the determination of phytochemicals in the crude leaves, pericarp, seeds, bark and root extracts obtained by using polar and non-polar solvents. The disc diffusion and micro-dilution methods were used to evaluate the antimicrobial activities of the crude solvent extracts against 13 microorganisms. The insecticidal properties of the crude extracts on larvae of Bactrocera zonata (Diptera: Tephritidae), which caused important economic losses to local fruits were also investigated. Results: All the extracts from the different parts of the plant showed antimicrobial activity against most tested microorganisms. The polar solvents' extracts of the fully mature parts of the castor plant were active against the Pseudomonas aeruginosa (ATCC 27853), Escherichia coli (E. coli) (ATCC 25922), E. coli (0145:H28 Acc. No. CP006027.1) with inhibition zones ranging from 16 mm to 19 mm and against Bacillus cereus (ATCC 11778) (B. cereus), Listeria innocua (ATCC 33090) (L. innocua). Lowest microbial inhibitory concentration was recorded for B. cereus, L. innocua, Staphylococcus aureus (ATCC 29213), E. coli (ATCC 25922) and Proteus mirabilis strain (NCTC 11938) with value of 3.2 mg/mL. The most active extract against both Gram-negative and Grampositive bacteria was extracted from the fully mature pericarps and it was the most active against E. coli (ATCC 25922), B. cereus, L. innocua and Staphylococcus aureus (ATCC 29213). In addition, the extracts obtained by using polar solvent and fully mature leaves demonstrated the strongest larvicidal activity against Bactrocera zonata (100%). Conclusions: Ricinus communis (castor) plant extracts possess larvicidal properties providing an effective eco-friendly control for fruit flies. The antimicrobial results justify the use of this plant in traditional medicine and the practice of supplementing decoctions/ concoctions with conventional antibiotics.

114-118

Authors: Lejan Miguel Alabastro Villareal, Rachelle Anne Montes Cruz, Michael Bagui Ples, Rodel Jonathan Santos Vitor II

Objective: To identify the effects of the consumption of non-nutritive sweeteners on memory retention and on the histology of the hippocampus. Methods: In this study, 20 mice were used to determine if there is an effect of consuming the maximum allowable dose of the non-nutritive sweeteners on the memory retention and on the histology of the hippocampus. The mice were distributed into four groups and the treatments were given via oral gavage: Group 1 (water), Group 2 (aspartame: 1 000 mg/kg), Group 3 (stevia: 1 000 mg/kg) and Group 4 (sucralose: 16 000 mg/kg). Treatments were administered to the different experimental groups for 32 days, after which memory retention was tested using the two-day water maze protocol. After the tests, the mice were sacrificed and the brain was analyzed histologically for neurotrophic effects. Results: Based on the results of the two-day water maze protocol, there were no differences between the non-nutritive sweeteners and the control group. However, stevia showed high cellular apoptosis followed by aspartame, sucralose and control group. Conclusions: There was no significant effect on the memory of the mice. It showed histologically however, that stevia had a significant neurotropic effect compared to the other sweeteners.

125-131

Authors: Miah Mohammed Abdus Satter, Mohammed Murtaza Reza Linkon Khan, Syeda Absha Jabin, Nusrat Abedin, Mohammed Faridul Islam, Badhan Shaha

Objective: To evaluate the nutritional composition, including major minerals, essential trace elements and toxic heavy metals of five different wild vegetables Dhekishak (Dryopteris filix-mas), Helencha (Enhydra fluctuans), Kalmishak (Ipomoea aquatica), Patshak (Corchorus capsularis) and Shapla stem (Nymphaea stellata) and their safety aspects. Methods: Proximate parameters moisture, ash, fat, fiber, protein, carbohydrate and energy; major minerals Na, K, Ca and Mg; trace elements Fe, Zn and Cu; and toxic heavy metals Pb, Cd, Cr, Ni and Hg were evaluated in the selected wild vegetables using the standard food analysis techniques. Results: The results from nutritional analysis showed that all the wild vegetables used in this study had a low content of crude fat and high content of moisture, ash, crude protein, crude fiber, carbohydrate and energy having the recommended dietary allowances. The vegetables were also rich in major minerals Na, K, Ca and Mg, sufficient in essential trace elements Fe, Cu and Zn while the heavy metals Pb, Cr and Ni were detected higher in amount in all the vegetables except Patshak than the limits recommended by Food and Agriculture Organization/World Health Organization. The heavy metals Cd and Hg were not detected in any vegetable. Conclusions: The outcome of this study suggests that the wild vegetables have very good nutritional potential to meet the recommended dietary allowances, but special awareness should be taken for public health concern about the high level of Pb, Cr and Ni which exceed the Food and Agriculture Organization/World Health Organization recommended limits for the metals in vegetables.

143-147

Authors: Nurul Izzati Osman, Norrizah Jaafar Sidik, Asmah Awal

Objective: To induce callus from the medicinally valuable species, Barringtonia racemosa L. (B. racemosa) whereby the formation of callus is essential for micropropagation studies and in vitro plant secondary metabolites production. Methods: The callus induction potential in B. racemosa was assessed from endosperm explant cultured on different culture media and plant hormonal treatments. Lloyd and McCown's woody plant medium and Murashige and Skoog's medium were used in the study as culture media. On the other hand, various concentrations and combinations of 2,4-dichlorophenoxyacetic acid (1.0–2.0 mg/L) and kinetin (0.5–2.5 mg/L) had been incorporated in the culture media to exert the effects of auxin and cytokinin on callus induction. Results: From the present study, it was found that the profuse [(1.681 ± 0.770) g fresh weight, (0.239 ± 0.239) g dry weight] and friable callus formation was optimally produced with desirable morphology and considerable percentage of callus induction (56.70%) in endosperm explants cultured on 1.0 mg/L 2,4-dichlorophenoxyacetic acid and 1.5 mg/L kinetin in Murashige and Skoog's medium. Conclusions: A reliable protocol for inducing callus formation of profuse and friable morphology in endosperm explant of B. racemosa had therefore been successfully established.

155-158

Authors: Faizal Rajeeb Mangudadatu Hussin, Rodel Jonathan Santos Vitor II, Julie Ann Oraa Joaquin, Melody Mendoza Clerigo, Anamy Ma. Caterial Paano

Objective: To examine the anti-hyperglycemic effects of aqueous Lenzites betulina (L. betulina) extracts on normoglycemic glucose-loaded mice. Methods: Different doses of aqueous extract from L. betulina were administered to 45 ICR mice (Mus musculus) to determine whether there was an effect ofL. betulina extracts on the blood glucose level of the ICR mice. Aqueous extracts of L. betulina were orally gavaged to mice using oral glucose tolerance test. A total of five groups were used to determine the effect of the fungi on blood glucose of the mice. Group A (positive control) was given 16.7 mg/kg glimepiride; Group B (negative control) was given distilled water; Group C (low dosage) was given 200 mg/kg aqueous extract; Group D (mid dosage) was given 400 mg/kg aqueous extract and Group E (high dosage) was given 800 mg/kg aqueous extract. Baseline blood glucose value was firstly acquired before induction of hyperglycemia through D-glucose, after which another check on blood glucose was made after 0.5 h. Immediately, after the acquisition of hyperglycemic blood glucose level, the individual administration of treatments were done. After that, three blood collections were done spanning 3 h with 1 h interval. Results: The low dose (200 mg/kg) and the mid dose (400 mg/kg) of L. betulina extracts were significantly different (P < 0.05) from their respective baseline values throughout the whole experiment with the latter surpassing its baseline value during the 3rd hour. On the other hand, the high dose (800 mg/kg) during the 1st hour after administration was not significantly different (P > 0.05) from its corresponding baseline value, acting faster than the positive control (glimepiride), which only became significantly different (P < 0.05) at the 2nd hour. Conclusions: Aqueous L. betulina extract is able to produce hypoglycemic effects on the mice with all doses, which are able to normalize blood glucose levels at varying times.

164-172

Authors: Ivan Viktorovich Borozda, Nikolay Alexandrovich Ganzhurov, Alexander Alexandrovich Kapustyansky, Roman Valerievich Nikolaev, Kirill Sergeevich Golokhvast

Objective: To evaluate the treatment outcomes of patients with pelvic ring injury by applying step-by-step external pelvic fixation and circular external fixation device. Methods: A total of 28 patients suffering from disintegrated pelvic ring injury are involved in the study. Fourteen patients (the treatment group) underwent step-by-step external pelvic fixation by applying anterior (anti-shock) and posterior modules. For the rest 14 patients (the experimental group), the osteosynthesis were conducted by means of a circular external fixation device. The long-term outcomes were evaluated in a year after the injury. Results: The residual deformity of 5 (4–7) mm was observed in 10 patients (71.4%) from the experimental group. In the treatment group, the residual deformity was evident only in 4 (28.6%) cases being 2.5 (2–3) mm (P = 0.000 319) on the average. The functional result (according to the Majeed scale) was statistically better in the treatment group (P = 0.000 319). Nine (64.3%) and five (35.7%) patients in treatment group showed excellent and positive results, respectively. The excellent result was demonstrated by 3 patients (21.4%) of the experimental group, the positive outcomes were observed in 6 cases (42.9%) and the unsatisfactory one was displayed by 1 patient (7.1%) of the same group. Conclusions: The modular approach applied is the advantage of the transosseous osteosynthesis allowing for a separate anterior (anti-shock) fixation and final posterior reposition of the pelvic ring preceded by the stabilization of vital functions. The above mentioned method gives an opportunity to increase the amount of techniques applied for the pelvic external fixation in polytrauma cases.

179-180

Authors: Beuy Joob, Viroj Wiwanitkit

No

8-15

Authors: Meriem El Kolli, Hocine Laouer, Hayet El Kolli, Salah Akkal, Farida Sahli

Objective: To evaluate the essential oils (EO) composition, antimicrobial and antioxidant power of a local plant, Daucus gracilis (D. gracilis). Methods: The aerial parts of D. gracilis were subjected to hydro distillation by a Clevenger apparatus type to obtain the EO which had been analyzed by gas chromatography and gas chromatography coupled with mass spectrometry, and screened for antimicrobial activity against five bacteria and three fungi by agar diffusion method. The mechanism of action of the EO was determined on the susceptible strains by both of time kill assay and lysis experience. The minimal inhibitory concentrations were determined by agar macrodilution and micro-dilution methods. Anti-oxidative properties of the EO were also studied by free diphenyl-2-picrylhydrazyl radical scavenging and reducing power techniques. Results: The EO yielded 0.68 (v/w). The chemical analysis presented two dominant constituents which were the elemicin (35.3%) and the geranyl acetate (26.8%). D. gracilis EO inhibited the growth of Bacillus cereus and Proteus mirabilis significantly with minimal inhibitory concentrations of 17.15 mg/mL by the agar dilution method and 57.05 mg/mL and 114.1 mg/mL, respectively by liquid micro-dilution. A remarkable decrease in a survival rate as well as in the absorbance in 260 nm was recorded, which suggested that the cytoplasm membrane was one of the targets of the EO. The EO showed, also, important anti-oxidative effects with an IC50 of 0.002 mg/mL and a dosedependent reducing power. Conclusions: D. gracilis EO showed potent antimicrobial and anti-oxidative activities and had acted on the cytoplasm membrane. These activities could be exploited in the food industry for food preservation.

20-25

Authors: Gbago Onivogui, Rebaone Letsididi, Mohamed Diaby, Liping Wang, Yuanda Song

Objective: To evaluate the influence of extraction solvents on antioxidant and antimicrobial activities of the pulp and seed of Anisophyllea laurina R. Br. ex Sabine fruits. Methods: The antibacterial activities of pulp and seed extracts were tested by using disk diffusion method against eight bacterial strains and three fungal strains. Total phenolic, flavonoid, monomeric anthocyanin and tannin contents, and antioxidant activities were determined by spectrometric methods. Results: The antioxidant analysis of pulp extract revealed the strong radical scavenging capacity and total phenolic content (4 329.66 mg of gallic acid/100 g), while seed extract showed the high antioxidant activity and total tannin content (5 326.78 mg catechin equivalent/100 g). Antibacterial and antifungal activities of methanol and ethanol extracts exhibited potent growth inhibitory activity against Aeromonas hydrophila, Bacillus subtilis, Escherichia coli O157:H7, Pseudomonas aeruginosa, Salmonella Typhimurium and Staphylococcus aureus ATCC 6538 with minimum inhibitory concentration values ranged from 125 to 250 mg/mL. However, seed extract had the strongest potential activity against Aspergillus niger and Candida albicans with minimum inhibitory concentration value of 500 mg/mL compared to pulp extract. Conclusions: Our results therefore demonstrated that ethanol and methanol extractions were more efficient in extracting antioxidants and bioactive compound in pulp and seed. These results support that these plant extracts can be used for the treatment of bacterial infections.

32-37

Authors: Fadila Moussaoui, Tajelmolk Alaoui

Objective: To demonstrate the in vitro antibacterial properties of five essential oils against ten bacterial strains and study the synergistic effect of the combination of essential oils with standard antibiotics. Methods: Origanum compactum, Chrysanthemum coronarium, Thymus willdenowii Boiss, Melissa officinalis and Origanum majorana L. were used alone and combined used with standard antibiotics to evaluate their antimicrobial activities. The disk diffusion method was employed. Results: The results showed that the combined application of the essential oils of the plants with antibiotics led to a synergistic effect in some cases, but antagonistic effect was also observed in some bacteria. Conclusions: This study shows that the combination of essential oils of the five plants with antibiotics may be useful in the fight against emerging microbial drug resistance.

44-49

Authors: Gehad Abdel Raheem Abdel Jaleel, Heba Mohammed Ibrahim Abdallah, Nawal E.L. Sayed Gomaa

Objective: To investigate some pharmacological effects including gastroprotective, antiinflammatory, analgesic, antipyretic and in vitro antioxidant effects of Artemisia herbaalba extract in different experimental models. Methods: Inflammation was induced in rat paw by subcutaneous injection of 1% (v/v) carrageenan solution. Writhes was induced in mice by intraperitoneal injection of 0.6% (v/v) acetic acid solution. Pyrexia was induced using Brewer's yeast suspension. Gastric lesion was induced in rats by oral administration of 99% ethanol. The anti-inflammatory, analgesic, antipyretic and gastroprotective activities of Artemisia herba-alba extract were investigated respectively. In vitro antioxidant effect was investigated using DPPH free radical. Results: The plant extract showed anti-inflammatory effect in carrageenan-induced paw edema in rats, analgesic effect against acetic acid-induced writhing, and antipyretic activity in Brewer's yeast model of pyrexia. Besides, it was shown to be a gastroprotective agent against ethanol-induced gastric ulcers. The plant also exhibited a free radical scavenging potential in an in vitro antioxidant study using DPPH. Conclusions: The results validate the use of the investigated plant in traditional medicine for different ailments.

55-59

Authors: Noor Azwani Mohd Rasidek, Mariam Firdhaus Mad Nordin, Kamyar Shameli

Objective: To study the jelly formulation produced by Musa acuminata Colla (AAA Group) peels and evaluate its antioxidant properties which are related to the product quality. Methods: The formulations of peel jelly were established under two-level full factorial designs within two blocks and one center point. Regarding response optimizer, the amount of sugar and citric acid was obtained; hence, the peel jellies were produced. The evaluation of antioxidant properties was conducted by using total phenolic content (TPC) assay and 1,1 diphenyl-2-picrylhydrazyl (DPPH) free radical assay. Results: The TPC of peel powder varied from 91.8 to 602.26 mg gallic acid equivalents/ 100 g dry weight, and 5%–7% peel jellies had phenolic content ranging from 29.38 to 48.31 mg gallic acid equivalents/100 g dry weight. The results of DPPH test indicated that at 10 mg/mL, the peel powder showed 89% DPPH inhibition, while 7% peel jelly prominently exhibited 84% DPPH inhibition. The correlation between DPPH IC50 value and TPC of peel powder as well as peel jelly was quite reasonably high with correlation coefficient ranging from 0.843 7 to 0.995. Conclusions: TPC can be used as an indicator in assessing the antioxidant activity of fruits and vegetables. The present investigation reveals that TPC is mainly responsible for DPPH free radical scavenging capacity.

67-70

Authors: Waleed Ibrahem Jalil, Mohammad Mushgil Zenad

Objective To highlight the presence of aerobic bacteria in live ticks infested sheep, in Diyala Governorate, Iraq. Methods One hundred and thirty adult alive ticks were picked up from sheep which were reared in different farms in Diyala Governorate, Iraq, during the period from November 2012 to May 2013. Ticks were classified in the Natural History Museum in Baghdad. They were dissected aseptically for extraction of the salivary gland and mid-gut. The removed tissue from each organ was inoculated in buffer peptone water (1%) and incubated for 2 h at 37 °C, to maintain weak and/or injured bacterial cells, then transmitted to nutrient broth incubated at 37 °C for 18 h. Culturing was done on three solid bacteriological media (nutrient, blood and McConkey agars), and then incubated at 37 °C for 24 h. Bacterial identification was performed by using multiple biochemical tests and API-20 strips. Data were analyzed by using Statistical Analysis System version 9.1, 2010. Chi-square test was used for comparison at significant level of P ≤ 0.05. Results Two species of ticks were identified [Rhipicephalus (Boophilus) annulatus and Hyalomma turanicum]. High bacterial isolation rate was observed (483 isolates). A significant high isolation rate was recorded from Rhipicephalus annulatus (63.14%). Six bacterial species were identified [Escherichia coli (28.36%), Pseudomonas aeruginosa (18.01%), Bacillus cereus (14.69%), Staphylococcus aureus (13.66%), Citrobacter freundii (13.04%), and Enterobacter species (12.21%)]. Also the high bacterial isolation rates were recorded in the temperate months (November, March and April); these coincided with high reproductive performance of ticks. Conclusions The high isolation rate of aerobic pathogens from ticks might reflect the active contribution of this arthropod in environmental contamination and increase the probability of transmitting bacterial pathogens to their hosts.

76-79

Authors: Lyudmila Vasilievna Veremchuk, Vera Innokentievna Yankova, Tatyana Isaakovna Vitkina, Anna Vladimirovna Nazarenko, Kirill Sergeevich Golokhvast

Objective To study the mechanism of formation of air quality and to determine the impact of the studied factors on asthma morbidity in Vladivostok. Methods The evaluation of air pollution in Vladivostok was done using long-term (2008–2012) monitoring data (temperature, humidity, atmospheric pressure, wind speed, etc.). The levels of suspended particulate matter, nitrogen and sulfur dioxide, carbon monoxide, ammonia, formaldehyde (mg/m3) in six stationary observation posts were assessed. We studied the aerosol suspensions of solid particles, which were collected during snowfall from precipitation (snow) and air in 14 districts with different levels of anthropogenic impact. Melted snow was analyzed on laser granulometry. The impact of air pollution on the distribution of asthma morbidity was evaluated in various age groups by data of federal statistical observation obtained from 8 adults and 7 children municipal clinics in Vladivostok (2008–2012). Results The content of suspended particulate components of pollution remained more stable, due to the features of atmospheric circulation, rugged terrain and residential development. The nano- and micro-sized particles (0–50 μm), which can absorb highly toxic metals, prevail in dust aerosols. These respirable fractions of particles, even in small doses, can contribute to the increase in asthma morbidity in the city. Conclusions We determined that asthma morbidity depends from general air pollution (in the range of 18.3%). It was detected that the highest age-specific dependence is associated with the content of particulate matter, carbon monoxide and nitrogen dioxide in air.

84-89

Authors: Angus Nnamdi Oli, Callistus Chibuike Ekejindu, David Ufuoma Adje, Ifeanyi Ezeobi, Obiora Shedrack Ejiofor, Christian Chibuzo Ibeh, Chika Flourence Ubajaka

Objective To assess healthcare workers' involvement in healthcare waste management in public and private hospitals. Methods Validated questionnaires (n = 660) were administered to randomly selected healthcare workers from selected private hospitals between April and July 2013. Results Among the healthcare workers that participated in the study, 187 (28.33%) were medical doctors, 44 (6.67%) were pharmacists, 77 (11.67%) were medical laboratory scientist, 35 (5.30%) were waste handlers and 317 (48.03%) were nurses. Generally, the number of workers that have heard about healthcare waste disposal system was above average 424 (69.5%). More health-workers in the government (81.5%) than in private (57.3%) hospitals were aware of healthcare waste disposal system and more in government hospitals attended training on it. The level of waste generated by the two hospitals differed significantly (P = 0.008 6) with the generation level higher in government than private hospitals. The materials for healthcare waste disposal were significantly more available (P = 0.001) in government than private hospitals. There was no significant difference (P = 0.285) in syringes and needles disposal practices in the two hospitals and they were exposed to equal risks (P = 0.851 0). Fifty-six (18.5%) and 140 (45.5%) of the study participants in private and government hospitals respectively were aware of the existence of healthcare waste management committee with 134 (44.4%) and 19 (6.2%) workers confirming that it did not exist in their institutions. The existence of the committee was very low in the private hospitals. Conclusions The availability of material for waste segregation at point of generation, compliance of healthcare workers to healthcare waste management guidelines and the existence of infection control committee in both hospitals is generally low and unsatisfactory.

982-986

Authors: Prakash Bangalore Nagendrappa, Jean-François Franetich, Frederick Gay, Audrey Lorthiois, Padma Venkatasubramanian, Dominique Mazier

Objective: To evaluate the potential prophylactic activity of traditional polyherbal remedy against malaria. Methods: A traditional polyherbal remedy against malaria from Odisha, India was evaluated for its potential prophylactic activity using in vitro hepatic cell lines assay and the murine malaria system Plasmodium yoelii yoelii/Anopheles stephensi. Results: The polyherbal extract inhibited the Plasmodium yoelii hepatic stages in vitro (IC50 0.74 mg/mL), a therapeutic index of 9.54. In mice treated with the aqueous extract (2 000 mg/kg/day), peak parasitaemia values were 81% lower in the experimental 2.35% ± 0.14% as compared to controls 12.62% ± 0.52% (P < 0.001), suggesting significant prophylactic activity. Conclusions: The observations provide a proof of concept for a traditional malaria prophylactic remedy used by tribal populations in India.

992-995

Authors: Nermin Adel El Semary, Manar Fouda

Objective: To assess the anticancer activity of eight cyanobacterial hydrophilic extracts on Ehrlich ascites carcinoma cell line. Methods: The cyanobacterial strains used in the investigation were collected from diverse habitats in Egypt. The initial cytotoxicity test of cyanobacterial hydrophilic extracts was carried out by MTT assay. The in vitro anticancer activity of the four most active extracts was performed on MCF-7 cells using sulforhodamine B assay. Morphological and molecular techniques were used to characterise identity of the isolate from which the most potent cytotoxic extract was obtained. Results: Extracts from four cyanobacterial strains had higher cytotoxic activities scoring 76.68%, 77.70%, 76.70% and 74.45%, respectively. A considerable anticancer effect was only detected when the concentrated extracts were used. One cyanobacterial extract gave the highest anticancer activity on human breast adenocarcinoma cell line (57.6% of inhibition) as compared to control. The isolate was best-matched to Cyanothece sp. with sequence resemblance 98% to Cyanothece sp. strain PCC7564 and the phylogenetic analysis confirmed its close identity to the Cyanothece genus. Conclusions: This is the first study to report the anticancer effect of aqueous extracts derived from the unicellular Cyanothece sp. from Egypt and its potential as a plausible candidate for future mass biotechnological applications.

1005-1009

Authors: Heba Sedrak, Noaman El-Garem, Mervat Naguib, Heba El-Zawahry, Mohamed Esmat, Lila Rashed

Objective: To evaluate vascular endothelial growth factor (VEGF) levels in hepatocellular carcinoma patients before and after transcatheter arterial chemoembolization (TACE) and percutaneous ethanol injection (PEI) and its relation to treatment response. Methods: A total of 40 patients with unrespectable hepatocelluar carcinoma were assessed clinically. Twenty patients were suitable to be treated by TACE, while other 20 patients were treated with PEI. Serum VEGF levels were measured before and 1 month after each procedure by ELISA. Response was assessed after 1 month according to Union Internationale Contre le Cancer evaluation criteria based on change in tumor size as measured by ultrasound. Results: There was no significant difference between TACE and PEI groups with regard to age, sex, tumor size, response to local therapy, or VEGF and alpha-fetoprotein before and after therapy. VEGF levels after TACE were significantly higher than before TACE [(298.1 ± 123.6) pg/mL vs. (205.8 ± 307.3) pg/mL; P = 0.001]. Also, VEGF levels were significantly higher after PEI than before PEI [(333.8 ± 365.6) pg/mL vs. (245.3 ± 301.8) pg/mL; P = 0.000]. Non-responders of both groups had significantly high VEGF levels than responder's, both before [(985.0 ± 113.2) pg/mL vs. (117.1 ± 75.3) pg/mL; P < 0.001] and after therapy [(1 330.6 ± 495.7) pg/mL vs. (171.0 ± 94.7) pg/mL; P = 0.000)]. Conclusions: Both TACE and PEI were associated with an increase in serum VEGF in hepatocelluar carcinoma patients. Higher levels of VEGF before and after therapy were found in non-responders, suggesting that VEGF is a useful marker in predicting treatment response.

1018-1026

Authors: Latifah Saiful Yazan, Yong Sze Ong, Nur Elena Zaaba, Razana Mohd Ali, Jhi Biau Foo, Yin Sim Tor

Objective To determine the anti-breast cancer activities and the safety oral consumption of Dillenia suffruticosa root aqueous extract (DRAE) in BALB/c mice. Methods In the anti-breast cancer study, female BALB/c mice were divided into five groups (n = 12), which were (1) positive control (with breast cancer, untreated), (2) negative control (without breast cancer, untreated) and other three groups of mice with breast cancer treated with 1 000, 500 and 250 mg/kg of DRAE, respectively, by oral gavage for 28 days. All mice except from the negative control group were injected into the mammary fat pad with 4T1 cells (1 × 105 4T1 cells/0.1 mL of phosphate buffer solution). DRAE was administered orally on Day 11 after the tumor has developed. Results The tumor volume of the 1 000 mg/kg of DRAE group reduced significantly compared to the positive control while treatment with 500 mg/kg of DRAE had significantly inhibited metastasis to the heart. In the acute toxicity study, treatment with up to 5 000 mg/kg of DRAE was not toxic to the animals, indicating its safety when a large amount of this plant extract was ingested. Based on the sub-acute toxicity study, treatment of the highest dose of DRAE (1 000 mg/kg) had mild liver toxicity indicated by mild focal hemorrhage. Conclusions DRAE possesses anti-breast cancer properties but at the same time it shows mild toxicity to the liver. The non observable adverse effect dose for DRAE is 500 mg/kg.

1033-1036

Authors: Moniruzzaman, Shahinuzzaman, Ahsanul Haque, Rahima Khatun, Zahira Yaakob

Objective To evaluate the antibacterial activity of essential oil from Trigonella foenum-graecum seeds powder, and identify the compounds from the extracted oil. Methods The seeds powder of Trigonella foenum-graecum was subjected to Clevenger extractor. Seven strains of bacteria were used to test antibacterial activity of the extract. The activity against bacteria was tested by disk diffusion method using Whatman No. 1 filter paper. Gas chromatography mass spectrometry analysis was performed with an Agilent7890/5975B-gas chromatography/mass selective detector. Results The hydrodistillation of seeds powder yielded 0.285% (v/w) of oil. Disk diffusion of the oil showed bactericidal activity against both Gram negative and Gram positive bacteria of tasted strains. The inhibition zone ranged from (8 ± 0) mm to (15.0 ± 0.7) mm depending on microbial strains. Gas chromatography mass spectrometry analysis showed 14 different compounds. The total compounds represented 80.96% of the oil. Conclusions The antibacterial activity is due to the effects of different biological active compounds present in the extract. Identification of the compounds may help to develop new effective antimicrobial agent(s). Further researches on purification, characterization and toxicology of the active compounds are needed.

1042–1045

Authors: Ananya Dechakhamphu, Nattapong Wongchum

Objective To screen the effect of 28 medicinal plants on inhibition of pancreatic lipase and evaluate the phytochemical contents of extracts. Methods The ethanolic extracts of 28 traditional Thai herbal medicines were assayed for their in vitro activities against porcine pancreatic lipase using p-nitrophenyl butyrate as a substrate. Quantitative estimation of flavonoids, phenolics, and alkaloids was done. Results Extracts from four herbs, Memecylon edule Roxb., Garcinia vilersiana Pierre, Cryptolepis elegans Wall. and Phyllanthus chamaepeuce Ridl., at a concentration of 100 μg/mL, strongly inhibited porcine pancreatic lipase by 90.97%, 92.04%, 94.64% and 95.38%, respectively. There was a significant positive correlation between phenolic content and inhibition activity. Inhibition activity was significantly correlated with flavonoid and with alkaloid contents. Conclusions From this result, it could be concluded that herbs represent a rich of anti-pancreatic lipase compounds, in particular, Cryptolepis elegans Wall. and Phyllanthus chamaepeuce Ridl. It is suggested that the phytochemical compounds from these plants may be applied for the prevention and treatment of obesity or hyperlipidemia.

1051-1054

Authors: Mücahit Görük, Mehmet Sinan Dal, Tuba Dal, Abdullah Karakus, Recep Tekin, Nida Özcan, Orhan Ayyildiz

Objective To evaluate the febrile neutropenic patients with hematological malignancies hospitalized in hematology clinic with poor hygiene standards. Methods A total of 124 patients with hematological malignancies (69 male, 55 female) hospitalized in hematology clinic with poor hygiene conditions depending on hospital conditions, between January 2007 and December 2010, were evaluated, retrospectively. Results In this study, 250 febrile neutropenia episodes developing in 124 hospitalized patients were evaluated. Of the patients, 69 were men (56%) and 55 women (44%). A total of 40 patients (32%) had acute myeloid leukemia, 25 (20%) acute lymphoblastic leukemia, 19 (15%) non-Hodgkin's lymphoma, 10 (8%) multiple myeloma, and 8 (8%) chronic myeloid leukemia. In our study, 56 patients (22%) were diagnosed as pneumonia, 38 (15%) invasive aspergillosis, 38 (15%) sepsis, 16 (6%) typhlitis, 9 (4%) mucormycosis, and 4 (2%) urinary tract infection. Gram-positive cocci were isolated from 52% (n = 20), while Gram-negative bacilli 42% (n = 16) and yeasts from 6% (n = 2) of the sepsis patients, respectively. The most frequently isolated Gram-positive bacteria were methicillin-resistant coagulase-negative staphylococci (n = 18), while the most frequently isolated Gram-negative bacteria was Escherichia coli (n = 10). Conclusions Febrile neutropenia is still a problem in patients with hematological malignancies. The documentation of the flora and detection of causative agents of infections in each unit would help to decide appropriate empirical therapy. Infection control procedures should be applied for preventing infections and transmissions.

1060-1064

Authors: Mohammad Hassan Davami, Morteza Pourahmad, Rasoul Baharlou, Abdolreza Sotoodeh Jahromi, Abbass Ahmadi Vasmejani, Kavous Solhjoo, Hamid Reza Fallah, Mohsen Kalantari

Objective To identify the anti-Toxoplasma antibodies from blood donors who referred to blood transfusion bases of Jahrom County, using ELISA method. Methods Based on the prevalence and characteristics method, 400 serum samples were collected from blood donors referred to Jahrom blood transfusion bases, Southern Iran, during 2010–2011, designed at testing by ELISA. IgM and IgG antibodies against Toxoplasma gondii were tested using ELISA kits (Dia-Pro) on serums. The data were analysed by SPSS 19 software. Results Review of 400 cases, 54 of them were IgG positive for parasites (13.5%) and 346 of those with negative IgG (86.5%). In IgM examination, 1.75% of them have been positive IgM (7 cases) and 98.25% of them were IgM negative (393 cases). By comparing the different group ages, 40–50 year age group had the highest prevalence of IgG positive (17.9%) and the age group of 30–40 years had the highest incidence of IgM negative (2.5%). Conclusions Due to the serological infection rate of toxoplasmosis obtained from this study, toxoplasmosis should be considered as a significant transfusion risk factor in Jahrom and also in any region with similar situations.

889-895

Authors: Barbara Lamagna, Maria Pia Pasolini, Sandra Nizza, Karina Mallardo, Maurizio Formicola, Alessandro Costagliola, Gerardo Fatone, Filomena Fiorito, Orlando Paciello, Luisa De Martino

Objective To assess normal conjunctival cytological and bacteriological/fungal flora features in the Mediterranean buffalo (Bubalus bubalis). Methods Swabs were taken from the inferior conjunctival sac of both eyes of 57 healthy female buffaloes aged 24–36 months, with no evidence of ocular disease, farmed in Campania region (Southern Italy), for microbiological analysis. Conjunctival eye specimens of both eyes were subsequently obtained by a cyto-brush, for cytological analysis. The antimicrobial susceptibility of bacterial isolates was also determined using the disk-diffusion method on Mueller Hinton agar plates. Results Cytological examination of conjunctival swab specimens (114 eyes) revealed epithelial cells (basal, intermediate, columnar and superficial) in all samples, whereas neutrophils, lymphocytes and plasma cells were present in 70%, 10% and 2% of samples, respectively. Microorganisms, for a total of 261 aerobic bacteria and 6 fungi, were isolated from 112/114 conjunctival samples [98.25%; 95% confidence interval (CI): 93.18–99.70]. Only two conjunctival swabs did not yield bacteria and/or fungi (2/114, 1.75%; 95% CI: 0.30–6.82). Gram-positive aerobes were most commonly cultured (181/261, 69.35%; 95% CI: 63.31–74.81), with Enterococcus faecium and Staphylococcus lentus predominating. Escherichia coli was the most frequently isolated as Gram-negative bacteria (80/261, 30.65%; 95% CI: 25.19–36.69). The antimicrobial resistance patterns of the isolated bacteria showed amoxycillin/clavulanic acid and cephalothin as the least sensitive antibiotics for both Gram-positive and Gram-negative bacteria. Conclusions These results provided first information on normal conjunctival ocular microflora and cytological features in Mediterranean buffalo.

902-908

Authors: Yaser Hamadeh Tarazi, Ahmed Mahmoud Almajali, Mustafa Mohammad Kheer Ababneh, Humam Shawket Ahmed, Adnan Saleem Jaran

Objective To determine the prevalence, genetic relatedness, and pattern of antimicrobial susceptibility in methicillin-resistant Staphylococcus aureus (S. aureus) (MRSA) isolated from household dogs, farm dogs, and stray dogs, compared to isolates from their associated personnel. Methods MRSA was isolated from 250 nasal swabs (150 swabs from dogs and 100 swabs from humans). PCR assays were used to detect the presence of both the nuc and mecA genes, which confirmed the identity of S. aureus isolates and the presence of methicillin resistance, respectively. Disk diffusion was used to determine the antibiotic susceptibility against 15 antimicrobial agents along with an E-test that determined the minimum inhibitory concentration for oxacillin. Pulsed field gel electrophoresis was conducted to determine the genetic relatedness of MRSA isolates from dogs to those from associated and unassociated personnel. Results The prevalence of S. aureus in dogs and humans was 12.7% and 10.0% respectively, while the prevalence of MRSA isolates in dogs and humans was 5.3% and 5.0%, respectively. The prevalence of MRSA isolates in household dogs, farm dogs, and stray dogs was 7.8%, 4.7%, and 0.0%, respectively. MRSA isolates demonstrated a significantly higher rate of multi-resistance against three or more antimicrobial agents than methicillin-susceptible S. aureus (MSSA). Trimethoprim-sulphamethoxazole and chloramphenicol were the most effective antibiotics against all MRSA isolates. Pulsed field gel electrophoresis revealed a strong association between dog MRSA isolates and MRSA isolates from strongly associated personnel. Conclusions MRSA is prevalent in house dogs, as well as in dog rearing centers and among their strongly associated personnel. A strong association was found between the MRSA isolates from dogs and those from humans who are in close contact. In addition, MRSA isolates showed a high rate of multi-resistance compared to MSSA isolates.

915–920

Authors: I Wayan Suardana, Dyah Ayu Widiasih, I Gusti Ngurah Kade Mahardika, Komang Januartha Putra Pinatih, Budi Setiadi Daryono

Objective: To evaluate the zoonotic potency of Escherichia coli O157:H7 through arbitrarily primed-PCR (AP-PCR) methods as one of the DNA fingerprinting methods. Methods: A total of 14 isolates consisted of 11 isolates originated from human feces with renal failure symptoms, 2 isolates originated from cattle feces, and 1 control isolate were used in this study. DNA of each isolate was extracted, and their profiles were studied by using AP-PCR method with M13 F and M13 R arbitrary primers. Results: The results founded that all of 14 isolates had similarity range from 54.6% to 88.5%. Isolates KL-106(3) and KL-55(6) originated from humans showed the degree of similarity with isolates SM-25(1) and SM-7(1) originated from cattle as high as 85% and 77%, respectively. Conclusions: The high degree of similarity between isolates originated from cattle and human indicated the high potency of zoonoses. The results also concluded AP-PCR method as a briefly fingerprinting method in order to trace the epidemiological of E. coli O157:H7.

928–931

Authors: Gabriela Ulloa-Urizar, Miguel Angel Aguilar-Luis, Mar´ıa del Carmen De Lama-Odr´ıa, Jose Camarena-Lizarzaburu, Juana del Valle Mendoza

Objective: To evaluate the susceptibility of Pseudomonas aeruginosa (P. aeruginosa) in vitro to the ethanolic extracts obtained from five different Peruvian medicinal plants. Methods: The plants were chopped and soaked in absolute ethanol (1:2, w/v). The antibacterial activity of compounds against P. aeruginosa was evaluated using the cupplate agar diffusion method. Results: The extracts from Maytenus macrocarpa (“Chuchuhuasi”), Dracontium loretense Krause (“Jergon Sacha”), Tabebuia impetiginosa (“Tahuari”), Eucalyptus camaldulensis Dehn (eucalyptus), Uncaria tomentosa (“Uña de gato”) exhibited favorable antibacterial activity against P. aeruginosa. The inhibitory effect of the extracts on the strains of P. aeruginosa tested demonstrated that Tabebuia impetiginosa and Maytenus macrocarpa possess higher antibacterial activity. Conclusions: The results of the present study scientifically validate the inhibitory capacity of the five medicinal plants attributed by their common use in folk medicine and contribute towards the development of new treatment options based on natural products.

937–943

Authors: Robab Valizadeh, Ali Asghar Hemmati, Gholamreza Houshmand, Sara Bayat, Mohammad Bahadoram

Objective: To evaluate and practically demonstrate the influence of Althaea officinalis flower mucilage as a plant known in Iran's and other Middle Eastern countries' traditional medicine for its wound healing properties. Methods: Animals were divided into 6 groups of 5 cases including a non-treated group as the negative control group receiving no treatment, a group treated with eucerin as the positive control group, a phenytoin 1% group as a standard group treated topically with phenytoin 1% hand-made ointment, and treatment groups treated with hand-made Althaea officinalis flower mucilage (AFM) ointment in a eucerin base with different concentrations (5%, 10%, 15%). Results: Among the treatment groups, the AFM 15% ointment showed the best result. Wound healing duration was reduced by the surface application of these groups. Wound closure was completed on Days 14 and 15 in the AFM 15% ointment and phenytoin 1% groups, respectively. No significant difference was observed in healing period between these groups. Conclusions: In conclusion, AFM 15% ointment was found to reduce wound healing time without any significant difference with the phenytoin 1% ointment. The authors suggest increased AFM effectiveness in when combined with phenytoin or other effectual plants.

948–954

Authors: Husnul Khotimah, Mulyohadi Ali, Sutiman Bambang Sumitro, Mochamad Aris Widodo

Objective To observe the effects of Centella asiatica (C. asiatica) methanolic extract on α-synuclein aggregation and its expression in rotenone-exposed zebrafish. Methods Zebrafish (Danio rerio) were exposed to 5 μg/L rotenone for 28 days and co-incubated with 2.5, 5.0 and 10.0 μg/mL of C. asiatica methanolic extract. The medium was changed every 48 h for maintain the concentration of rotenone and extract. After 28 days zebrafish were sacrificed on the ice block and protein was isolated from zebrafish brain for ELISA of dopamine and Western blotting of α-synuclein. Immunohistochemistry was conducted to observe the α-synuclein expressions from histopathological preparation of zebrafish brain. The head were soaked in 10% formaline for less than 24 h and embedded onto paraffin block, then sliced for immunohistochemistry using anti α-synuclein antibody. We also measured zebrafish motility for 5 min in each week. Results C. asiatica has important bioactive compounds such as asiaticoside that has anti-inflammatory and antioxidant properties. It may inhibit cascade reaction due to oxidative stress induced by rotenone. Decreasing reactive oxygen species proposed probability of radical attack to α-synuclein protein that caused aggregation and increase of its expression. The motility of zebrafish was also maintained in C. asiatica groups due to the increasing dopamine level in rotenone-induced zebrafish. High level of reactive oxygen species inactivated enzyme for dopamine synthesis such as tyrosine hydroxylase, and oxidized dopamine itself. Oxidized dopamine increased α-synuclein aggregation. Thus, the dopamine level decreased in rotenone-induced zebrafish, but C. asiatica increased dopamine level. Conclusions C. asiatica has a potential to be developed as an anti-Parkinson's disease treatment due to its capability for minimized the sign of Parkinson's such as α-synuclein aggregation and expression, increasing motility and dopamine as well.

960–964

Authors: Tarek Benmeddour, Hocine Laouer, Salah Akkal, Guido Flamini

Objective To evaluate the essential oil composition and the antibacterial activity of an Algerian endemic plant, Launaea lanifera Pau (L. lanifera), grown in arid steppe regions. Methods L. lanifera essential oil was isolated from aerial parts by steam distillation and its chemical composition was evaluated by gas chromatography-flame ionization detector and gas chromatography with electron impact mass spectrometry. Furthermore, its in vitro antibacterial activity against four bacterial strains was tested following the agar disk diffusion method. Results This species had a very low essential oil yield (0.005%). Twenty-four (92.6%) individual components were identified. The main constituents were hexahydrofarnesyl acetone (31.6%), (E)-β-ionone (8.5%), (E)-β-damascenone (7.0%), 2-methyltetradecane (3.8%), n-heptadecane (3.8%), limonene (2.8%) and β-caryophyllene (2.8%). No noteworthy antimicrobial activity was observed on the tested bacteria, neither Gram negative nor Gram positive. Conclusions This is the first report on the volatile constituents and antibacterial activity of L. lanifera. The studied essential oil does not possess significant activity against the tested microorganisms.

971–973

Authors: Luis Antonio Gomez-Puerta, Armando Emiliano Gonzalez, Cesar Gavidia, Viterbo Ayvar, Hector Hugo Garcia, Maria Teresa Lopez-Urbina, for the Cysticercosis Working Group in Peru

The efficacy of oxfendazole (OFZ) on Taenia hydatigena metacestodes, also called Cysticercus tenuicollis (C. tenuicollis), was studied in 648 raising pigs. This study was performed in Tumbes Department in Peru, an endemic area for cysticercosis. Pigs were randomized in two groups; untreated group (n = 142) did not receive any treatment and treated group (n = 506) received OFZ treatment at a single dose of 30 mg/kg body weight. Six months after treatment, the pigs were necropsied. The prevalence of infection by C. tenuicollis among the pigs was 27.5% (39/142) and 2.0% (10/506) in untreated and treated groups, respectively. Untreated group was infested only with viable cysts, whereas treated group had no viable cysts. All the cysts found in treated group presented degeneration, with a thick membrane, and they contained milky fluid and fibrous tissue. A single dose of OFZ was effective against C. tenuicollis, thus providing an alternative drug for controlling this parasite in pigs.