Asian Pacific Journal of Tropical Biomedicine

847-852

Authors: Arief Nurrochmad, Endang Lukitaningsih, Ameilinda Monikawati, Dita Brenna Septhea , Edy Meiyanto

Objective To investigate the estrogenic effect of (8,9)-furanyl-pterocarpan-3-ol (FPC) on growth of human breast cancer T47D cells and the interactions between the FPC and tamoxifen (TAM), on the growth of estrogen receptor-dependent breast cancer T47D cells. Methods The proliferation effect of FPC were conducted on T47D cells in vitro by MTT test. T47D cells were treated with FPC alone (0.01-200 µmol/L) or in combination with TAM 20 nmol/L. Furthermore, the expression of ERα or c-Myc were also determined by immunohistochemistry. Results The results indicated that administration of an anti-estrogen TAM showed growth inhibitory effect on T47D cells, wheraes co-administered with low concentration (less than 1 µmol/L) of FPC attenuated to promote cell proliferation. In contrast, the combination of TAM with higher doses (more than 20 µmol/L) of FPC showed growth inhibitory. This result was supported by immunocytochemistry studies that the administration of 20 nmol/L TAM down-regulated ER-α and c-Myc, but the combination of 20 nmol/L TAM and 1 µmol/L FPC robustly up-regulated expression of ER-α. Thus, the reduced growth inhibition of TAM 20 nmol/L by FPC 1 µmol/L on T47D cells may act via the modulation of ER-α. Conclusions The findings indicate and suggest that FPC had estrogenic activity at low concentrations and anti-estrogenic effect that are likely to be regulated by c-Myc and estrogen receptors. We also confirm that low concentration of FPC attenuated the growth-inhibitory effects of TAM on mammary tumor prevention. Therefore, the present study suggests that caution is warranted regarding the consumption of dietary FPC by breast cancer patients while on TMA therapy.

859-865

Authors: Saiba Abdul Wahab Nusaiba, Kumaraswamy Murugan

Objective Natural products of plant origin are potential source of novel antimicrobial and antioxidative agents. Thottea siliquosa (Lam.) Ding Hou. (T. siliquosa). A medicinal herb used by local tribals for treating various ailments. The present study aims at the phytochemical screening, GC-MS analysis, in vitro antibacterial activity and antioxidant potentiality of root and leaf extracts of T. siliquosa. Methods Hot continuous Soxhlet extraction, GC-MS analysis, antibacterial analysis by disc diffusion, microdilution assay and antioxidant potentialities by hydroxyl radical and nitric oxide radical scavenging. The data was statistically analyzed. Results Phytochemical screening of the ethyl acetate and methanolic extract of leaf and root revealed the presence of phenols, alkaloids, tannins and saponin. The extract revealed a pool of phytochemicals by comparison with authentic standards from spectral library. Both the extracts has shown their broad spectrum of inhibition against the selected bacteria Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa and Klebsiella pneumonia compared with standard antibiotic drug streptomycin. The extracts showed antioxidant activity by scavenging of free radicals such as hydroxyl and nitric oxide. The IC50 values of the ethyl acetate extracts leaf and root and standard in this assay were 167.5±0.67, 99.4±1.2, 192±2.5 µg/mL respectively. Similarly those methanolic extracts of leaf and root were 269.5±0.89 and 289.1±2.66 µg/mL respectively. Similarly, ethyl acetate and methanolic extracts also caused a moderate dose-dependent inhibition of nitric oxide with an IC50 range 65.5±1.55 to 148 ±3.09 µg/mL. The inhibitory activities were found to be dose dependent. Conclusion The present study provides evidence that ethyl acetate and methanol extract of leaf and root of T. siliquosa are potential source of natural antioxidants and bactericidal nature. It is essential that research should continue to isolate and purify the bio active components of this natural plant and use in drug discovery and development.

871-876

Authors: Suprava Sahoo, Goutam Ghosh, Debajyoti Das, Sanghamitra Nayak

Objective To investigate antioxidant potential of methanol extract of Alpinia nigra leaves. Methods The study was done by using various in vitro methods such as 1,1-diphenyl-2-picrylhydrazyl (DPPH), 2,2-azino-bis(3-ethylbenzthiazoline-6-sulphonic acid) (ABTS), nitric oxide and hydrogen peroxide radical scavenging assays. Phytochemical constituents, total phenolic content and total flavonoid content of the extract at different concentrations (10-500 µg/mL) were determined. Results Alpinia nigra leaves showed high free radical scavenging activity as evidenced by the low IC50 values in DPPH (64.51 µg/mL), in ABTS (28.32 µg/mL), in nitric oxide (80.02 µg/mL) and in H2O2 (77.45 µg/mL) scavenging assays. Furthermore the TPC and TFC of the extract were found to be 69.25 mg gallic acid equivalent per gram of extract and 78.84 mg quercetin equivalent per gram of extract respectively. Conclusions The results of present comprehensive analysis demonstrated that Alpinia nigra leaves possess high phenolic, flavonoid contents and potential antioxidant activity, and could be used as a viable source of natural antioxidants and might be exploited for functional foods and neutraceutical applications.

882-886

Authors: Shivakumar Singh Policepatel, Vidyasagar Gunagambhire Manikrao

Objective To document traditional medicinal plants knowledge used in treating skin diseases at Hyderabad Karnataka Region. Methods The information on the use of medicinal plants in the treatment of skin diseases was gathered from traditional herbal healers and other villagers through interviews. Results A total of 60 plants species belonging to 57 genera and 34 families were found useful and herewith described them along with the method of drug preparation, mode of administration, probable dosage and duration of treatment. Several new findings on the traditional rural practices were reported. Conclusions The present study revealed that the Hyderabad Karnataka rural people is primarily dependent on medicinal plants for treating skin diseases.

896-901

Authors: Mahendra Kumar Verma, Francies Xavier, Yogendra Kumar Verma, Kota Sobha

Objective To isolate, partially purify and evaluate the cytotoxic and antitumor activity of a serine protease from the chosen Indian earthworm Pheretima posthuma. Methods Whole animal extract was prepared and purified its protein constituents by size and charge based chromatographic separation techniques using Sephadex G-50 and DEAE-Cellulose resin respectively. Average molecular weight of the protein isolate was determined and analyzed for its cytotoxic property against Vero cells in different dilutions (1: 20 and 1: 40) and anti-tumor activity by MTT assay (a colorimetric assay) using breast cancer cell line MCF-7, with tamoxifen as standard. Results One of the protein constituents after purification was characterized as serine protease by Caseinolytic plate diffusion assay. Average molecular weight of this purified isolate was determined, by SDS-PAGE analysis with standard protein ladder, as of 15 kDa. The performed tests suggested that the 15kDa fraction has potent cytotoxic activity and satisfactory antitumor activity as well in vitro. Conclusions Exact molecular mechanism of the cytotoxic and antitumor activities is yet to be explored and currently we are working on ultra-purification and biophysical characterization of this fraction. Further investigation into the mechanism(s) of cytotoxic and antitumor activities at molecular level would be useful in treatment of various classes of cancer and viral infections in future.

907-912

Authors: Dhumal Uttareshvar Mahaling, Madole Mahesh Basavaraj, Aher Jagdish Bika

Objective To detect and compare serum lipid abnormalities in patients diagnosed with different grades of non-alcoholic fatty liver on ultrasonography. Methods A total of 70 cases which included 30 males and 40 females, diagnosed as non-alcoholic fatty liver disease (NAFLD) on ultrasound were investigated with serum lipid profile. Then a comparison of lipid abnormalities between different grades of fatty liver diagnosed on ultrasound was done. P value was calculated by using analysis of variance test (ANOVA) and P value <0.05 was considered as statistically significant. Results Out of 70 cases which were diagnosed as NAFLD on ultrasonography, grade I NAFLD cases were 47.15%, grade II were 42.85% and grade III were 10%. The mean age of the patients was 49.14 years. Male to female ratio was 3:4. Serum triglycerides, total cholesterol, LDL and VLDL levels were raised in 67.14%, 45.71% 34.28%, 25.71% of cases respectively. Low serum HDL levels were seen in 62.85% of patients. On statistical analysis we found increasing grades of NAFLD were significantly associated with increasing values of total cholesterol (P value-0.001), LDL (P value-0.000) and VLDL (P value-0.003) and decreasing HDL (P value-0.000). Conclusion Most of the patients of NAFLD in India is asymptomatic, non-diabetic and non-hypertensive. Though liver biopsy is the gold standard method for diagnosis of NAFLD, Ultrasonography which is non-invasive, simple tool, can be used for the early detection of NAFLD in asymptomatic patients.

916-924

Authors: Nurul Fariza Rossle, Baha Latif

The protozoa under the genus Cryptosporidium is a zoonotic apicomplexan obligate intracellular parasite. Cryptosporidiosis, the term used to designate infection caused by Cryptosporidium sp., is considered as one of the most common food and waterborne diseases with worldwide spread, acting as a common cause of diarrhoea in animals and man. In immunocompetent individuals, Cryptosporidium typically induces self-limiting diarrhoea, which may resolve on its own after 2-3 d. However, cryptosporidiosis may turn life-threatening and subsequently lead to death in small children, the elderly and immunocompromised person, especially in AIDS patient. The diagnosis for Cryptosporidium infection is usually carried out through examination of stool for the presence of oocysts which measured 4-6 µm with spherical appearance. Morphometric identification is often difficult because of the diminutive size and obscure internal structure of the protozoa. Often, the identification of Cryptosporidium is realised through the combination of methods incorporating data from morphometrics, molecular techniques, and host specificity. However, limitations to some of these techniques still exist whether because of cost, duration, expertise, or reliability. Drugs combination is implemented in treatment of cryptosporidiosis. The efficiency of paromomycin, an aminocyclitol antibiotic isolated from Streptomyces, can be effective when combined use with protease inhibitors or recombinant IL-12. Since there is no drug that achieves the complete removal of Cryptosporidium from the host, supportive therapy was preferred in both human and domestic animals.

767-775

Authors: Fatemeh Kamiabi, Zairi Jaal, Chan Lai Keng

Objective To evaluate the growth inhibition activity of the crude extract of Cyperus aromaticus (C. aromaticus) cultured cells against the 3rd instar larvae of Aedes aegypti (Linn.) and Aedes albopictus Skuse (Ae. albopictus) under laboratory conditions, and determine the sublethal effects (EI50) of the crude extract of C. aromaticus cultured cells on some biological and morphological parameters of both Aedes mosquito species during two generations as well. Methods The cell suspension cultures of C. aromaticus were activated from five callus lines (P4, Pa, Z1, Z6 and Ml) derived from the root explants of in vitro plantlets. The cultured cells were extracted in chloroform and used as plant material for the present study. For detection of juvenile hormone III, the crude extracts were analyzed by HPLC. Then the crude extracts of the three C. aromaticus cultured cell lines which contained varied amounts of juvenile hormone III [high level (P4 cell line), medium level (Z1 cell line) and low level (Ml cell line)] were tested against Aedes mosquito species. Laboratory evaluation was performed against late third instar larvae of the Vector Control Research Unit strains of Ae. aegypti and Ae. albopictus using the standard WHO method. The effects of EI50 of the C. aromaticus cultured P4 cells on fecundity, fertility, growth period, sex ratio, adult size and longevity of Aedes mosquitoes were assessed. Results Bioassay tests presented the remarkable growth inhibition activity of the crude extracts of C. aromaticus cultured cells against the two Aedes mosquitoes. Between the two mosquito species, Ae. albopictus was more susceptible to the crude extracts with lower EI50 values. EI50 of the crude extract of C. aromaticus cultured cells (P4) increased the sterility indices in the parental generation females in both Aedes mosquito species. A significant delay in the pupal formation and adult emergence were observed in the parental generation of the both mosquito species. The sex ratio of the adult population either parental or F1 generation of the Aedes mosquito species was not significantly affected by the EI50 dosage of the crude extract of C. aromaticus cultured P4 cells. A significant decrease in the wing length of the treated adult (female and male) of Aedes aegypti as well as the treated female of Ae. albopictus were observed. Longevity of the adult female of the parental generation of both Aedes mosquitoes as well as females of F1 generation of Ae. albopictus were significantly decreased. Conclusions The present study revealed the potential of the crude extract of C. aromaticus cultured cells in controlling vector mosquito populations in the effort to reduce the transmission of vector borne diseases.

780-784

Authors: Fatiany Pierre Ruphin, Robijaona Baholy, Randrianarivo Emmanuel, Raharisololalao Amelie, Marie-Therese Martin, Ngbolua Koto-te-Nyiwa

Objective To validate scientifically the traditional use of Salacia leptoclada Tul. (Celastraceae) (S. leptoclada) and to isolate and elucidate the structure of the biologically active compound. Methods Bioassay-guided fractionation of the acetonic extract of the stem barks of S. leptoclada was carried out by a combination of chromatography technique and biological experiments in viro using Plasmodium falciparum and P388 leukemia cell lines as models. The structure of the biologically active pure compound was elucidated by 1D and 2D NMR spectroscopy and mass spectrometry. Results Biological screening of S. leptoclada extracts resulted in the isolation of a pentacyclic triterpenic quinone methide. The pure compound exhibited both in vitro a cytotoxic effect on murine P388 leukemia cells with IC50 value of (0.041±0.020) µg/mL and an antiplasmodial activity against the chloroquine-resistant strain FC29 of Plasmodium falciparum with an IC50 value of (0.052±0.030) µg/mL. Despite this interesting anti-malarial property of the lead compound, the therapeutic index was weak (0.788). In the best of our knowledge, the quinone methide pentacyclic triterpenoid derivative compound is reported for the first time in S. leptoclada. Conclusions The results suggest that furthers studies involving antineoplastic activity is needed for the development of this lead compound as anticancer drug.

790-797

Authors: Berhanu Andualem, Amare Gessesse

Objective To investigate and optimize microbial media that substitute peptone agar using brebra seed defatted flour. Methods 'Defatted process, inoculums preparation, evaluation of bacterial growth, preparation of cooked and hydrolyzed media and growth turbidity of tested bacteria were determined. Results Two percent defatted flour was found to be suitable concentration for the growth of pathogenic bacteria: Escherichia coli (ATCC 25922) (E. coli), Pseudomonas aeruginosa (ATCC 27853), Salmonella (NCTC 8385) and Shigella flexneri (ATCC 12022) (S. flexneri), while 3% defatted flour was suitable for Staphylococcus aureus (ATCC 25923) (S. aureus). E. coli (93±1) and S. flexneri (524±1) colony count were significantly (P≤0.05) greater in defatted flour without supplement than in supplemented medium. E. coli [(3.72×109±2) CFU/mL], S. aureus [(7.4×109±2) CFU/mL], S. flexneri [(4.03×109±2) CFU/mL] and Salmonella [(2.37×109±1) CFU/mL] in non-hydrolyzed sample were statistically (P≤0.05) greater than hydrolyzed one and commercial peptone agar. Colony count of Salmonella [(4.55×109±3) CFU/mL], S. flexneri [(5.40×109±3) CFU/mL] and Lyesria moncytogenes (ATCC 19116) [(5.4×109±3) CFU/mL] on raw defatted flour agar was significantly (P≤0.05) greater than cooked defatted flour and commercial peptone agar. Biomass of E. coli, S. aureus, Salmonella and Enterococcus faecalis in non-hydrolyzed defatted flour is highly increased over hydrolyzed defatted flour and commercial peptone broth. Conclusions The defatted flour agar was found to be better microbial media or comparable with peptone agar. The substances in it can serve as sources of carbon, nitrogen, vitamins and minerals that are essential to support the growth of microorganisms without any supplements. Currently, all supplements of peptone agar are very expensive in the market.

806-810

Authors: Nguyen Hoang Loc, Nguyen Thi Duy Nhat

Objective To investigate the effects of some culture conditions on production of asiaticoside from centella (Centella asiatica L. Urban) cells cultured in 5-L bioreactor. Methods The centell cell suspension culture was conducted in 5-L bioreactor to investigate the growth and asiaticoside accumulation under various conditions. Asiaticoside content was determined by HPLC analysis. Results The results showed that the cell growth and asiaticoside accumulation peaked after 24 d of culture at an agitation speed of 150 r/min and aeration rate of 2.5 L/min. The cell biomass reached a maximum value of 302.45 g fresh weight (31.45 g dry weight) and growth index of 3.03 with inoculum size of 100 g. However, asiaticoside content was the highest (60.08 mg/g dry weight) when culture was initiated with an inoculum size of 50 g. Conclusions The present study found the suitable conditions for growth of centella cells and their asiaticoside production in bioreactor.

816-821

Authors: Moussa Ahmed, Noureddine Djebli, Saad Aissat, Baghdad Khiati, Abdelmalek Meslem, Salima Bacha

Objective To evaluate the in vitro activity and synergism of the combinations of natural honey and curcuma starch against Rhodotorula mucilaginosa in correlation with total phenolic, flavonoid contents, and diastase activity. Methods The Folin-Ciocalteu test was used to determine the total polyphenols content and the flavonoid content was analyzed using by the aluminum chloride method. The antifungal activity of the natural honey, determined by an agar well diffusion assay and agar incorporation method. Results Total phenolic content varied from (63.93±0.11) to (95.36±6.08) mg GAE/100 g honey as gallic acid equivalent. Total flavonoids content varied from (5.41±0.04) to (9.94±0.54) mg CE/100 g. Diastase activity values were between (7.3±2.8) and (26±2.8). The zone inhibition diameter for the six honey samples without starch ranged between 6 and 20 mm. When starch was mixed with honey and then added to well, a zone inhibition increase diameter 7 and 21 mm. The percentage increase was noticed with each variety and it ranged between 5% and 62.5%. The minimal inhibitory concentrations for the six varieties of honey without starch against Rhodotorula mucilaginosa ranged between 28% and 36% (v/v). When starch was incubated with honey and then added to media, a minimal inhibitory concentration drop has been noticed with each variety. It ranged between 6.66 % and 20% (w/v). No significant correlation was established between diastase activity and bioactive compounds. Conclusions The mixture of curcuma starch and honey could lead to the development of new combination antibiotics against Rhodotorula infections

825-829

Authors: Rassi Yavar, Karami Hadi, Abai Mohammad Reza, M Mohebali, Bakshi Hasan, Oshaghi Mohammad Ali, Rafizadeh Sina, Bagherpoor Hagigi Habib, Hosseini Abodolrahim, Gholami Manuchehr

Objective To identify the vectors and reservoirs of cutaneous leishmaniasis in the endemic focus of Farashband, Fars Province, South of Iran. Methods Sticky papers and Sherman trap were used for collection of sand flies and rodents, respectively. Polymerase chain reaction (PCR) of kDNA, ITS1-rDNA were used for identification of Leishmania parasite in sand flies as well as rodents. Results Totally 2 010 sand flies were collected and the species of Phlebotomus papatasi Scopoli was the common specimen in outdoors and indoors places. PCR technique was employed on 130 females of Phlebotomus papatasi. One of them (0.76%) was positive to parasite Leishmania major (L. major) and one specimen (0.76%) was positive to Leishmania infantum. Microscopic investigation on blood smear of the animal reservoirs for amastigote parasites revealed 16 (44%) infected Tatera indica. Infection of them to L. major was confirmed by PCR against kDNA loci of the parasite. Conclusions The results indicated that Phlebotomus papatasi was the dominant species circulating two species of parasites including L. major and Leishmania infantum among human and reservoirs. Furthermore, Tatera indica is the only main host reservoir for maintenance of the parasite source in the area.

834-840

Authors: Khaled MM Koriem

Oleum azadirachti consists of the oil obtained from dried seeds of Azadirachta indica A. Juss. (family: Meliaceae). Local names of Azadirachta indica A. Juss. are Abodua, aforo-oyinbo, anwe egyane, arista, azad dirakht, azadarakht, azedarach and bead tree. Indigenous to India, and widely distributed in South and South-East Asia and cultivated in Africa, the South Pacific Islands, South and Central America and Australia, and in southern Florida and California, United States of America, it is a straight-boled deciduous tree, which is 6-25 m high. Bark is dark-brown, externally fissured with a buff inner surface and fibrous fracture. Leaves alternately arranged, pinnately compound and up to 40 cm long, and composed of 8-18 short-petiolate narrow-ovate, pointed and curved toothed leaflets, 3-10 cm long and 1-4 cm wide arranged in alternate pairs. The major constituents are oxidized tetranortriterpenes including azadirachtin (azadirachtin A), azadiriadione, epoxyazadiradione, azadirone, nimbidin, nimbin, deacetylnimbin, salannin, gedunin, mahmoodin, 17-hydroxydiradione and related derivatives. It is of various medicinal uses, such as a contraceptive for intravaginal use, a mosquito repellent, and treatment of vaginal infections, treatment of gastric ulcers, cardiovascular disease, malaria, rheumatism and skin disorders, external applications for treatment of septic wounds,ulcers and boils, treatment of allergic skin reactions, asthma, bruises, colic, conjunctivitis, dysmenorrhoea, fever, gout, headache, itching due to varicella, kidney stones, leukorrhoea, psoriasis, scabies, sprains and muscular pain, and wounds. It is also used as an emmenagogue, tonic, stomatic and vermicide. In conclusion, the plant oil had antifertility, antihyperglycaemic, anti-inflammatory, antimicrobial, antiviral, antiulcer, estrogenic, immune, contraceptive, antibacterial, insect repellent, and skin treatment effects.

678-681

Authors: Doaa M Abd El-Aziz

Objective To detect Salmonella typhimurium (S. typhimurium), one of the most frequently isolated serovars from food borne outbreaks throughout the world, in retail raw chicken meat and giblets. Methods One hundred samples of retail raw chicken meat and giblets (Liver, heart and gizzard) which were collected from Assiut city markets for detection of the organism and by using Duplex PCR amplification of DNA using rfbJ and fliC genes. Results S. typhimurium was detected at rate of 44%, 40% and 48% in chicken meat, liver and heart, respectively, but not detected in gizzard. Conclusions The results showed high incidence of S. typhimurium in the examined samples and greater emphasis should be applied on prevention and control of contamination during processing for reducing food-borne risks to consumers.

692-696

Authors: Kwan Yuet Ping, Ibrahim Darah, Yeng Chen, Sreenivasan Sasidharan

Objective To evaluate the cytotoxicity and genotoxicity activity of Euphorbia hirta (E. hirta) in MCF-7 cell line model using comet assay. Methods The cytotoxicity of E. hirta extract was investigated by employing brine shrimp lethality assay and the genotoxicity of E. hirta was assessed by using Comet assay. Results Both toxicity tests exhibited significant toxicity result. In the comet assay, the E. hirta extract exhibited genotoxicity effects against MCF-7 DNA in a time-dependent manner by increasing mean percentage of DNA damage. The extract of E. hirta showed significant toxicity against brine shrimp with an LC50 value of 620.382 µg/mL (24 h). Comparison with positive control potassium dichromate signifies that cytotoxicity exhibited by the methanol extract might have moderate activity. Conclusion The present work confirmed the cytotoxicity and genotoxicity of E. hirta. However, the observed toxicity of E. hirta extracts needs to be confirmed in additional studies.

705-710

Authors: Mohammad Amzad Hossain, Khulood Ahmed Salim AL-Raqmi, Zawan Hamood AL-Mijizy, Afaf Mohammed Weli, Qasim Al-Riyami

Objective To prepare various crude extracts using different polarities of solvent and to quantitatively evaluate their total phenol, flavonoids contents and phytochemical screening of Thymus vulgaris collected from Al Jabal Al Akhdar, Nizwa, Sultanate of Oman. Methods The leave sample was extracted with methanol and evaporated. Then it was defatted with water and extracted with different polarities organic solvents with increasing polarities. The prepare hexane, chloroform, ethyl acetate, butanol and methanol crude extracts were used for their evaluation of total phenol, flavonoids contents and phytochemical screening study. The established conventional methods were used for quantitative determination of total phenol, flavonoids contents and phytochemical screening. Results Phytochemical screening for various crude extracts were tested and shown positive result for flavonoids, saponins and steroids compounds. The result for total phenol content was the highest in butanol and the lowest in methanol crude extract whereas the total flavonoids contents was the highest in methanol and the lowest hexane crude extract. Conclusions The crude extracts from locally grown Thymus vulgaris showed high concentration of flavonoids and it could be used as antibiotics for different curable and uncurable diseases.

716-719

Authors: Ehsan Gharib Mombeni, Manoochehr Gharib Mombeini, Lucas Chaves Figueiredo, Luciano Soares Jacintho Siqueira, Debora Testoni Dias

Objective To determine the effects of high voltage electrical stimulation (HVES, 800 Voltage) on rapid decreases in pH values and improvements in meat quality. Methods A total of 50 beef carcasses were applied, divided into two groups, one as a control and another for HVES. Meat quality was evaluated based on M. longissimus dorsi by examining pH and temperature levels at 1, 2, 5, 10 and 24 h, while color stability was examined seven days after slaughter. Results HVES decreased the pH values of the meat and accelerated rigor mortis (P<0.05). HVES caused differences in instrumental color values compared with the control groups across the ageing period at 4 °C. Conclusion the HVES had positive effects on meat quality and color stability, in contrast to undesirable consumer preferences.

725-731

Authors: Berhanu Andualem

Objective To investigate the synergic antibacterial activity of garlic and tazma honey against standard and clinical pathogenic bacteria. Methods Antimicrobial activity of tazma honey, garlic and mixture of them against pathogenic bacteria were determined. Chloramphenicol and water were used as positive and negative controls, respectively. Minimum inhibitory concentration (MIC) and minimum bactericidal concentration of antimicrobial samples were determined using standard methods. Results Inhibition zone of mixture of garlic and tazma honey against all tested pathogens was significantly (P≤0.05) greater than garlic and tazma honey alone. The diameter zone of inhibition ranged from (18±1) to (35±1) mm for mixture of garlic and tazma honey, (12±1) to (20±1) mm for tazma honey and (14±1) to (22±1) mm for garlic as compared with (10±1) to (30±1) mm for chloramphenicol. The combination of garlic and tazma honey (30-35 mm) was more significantly (P≤0.05) effective against Salmonella (NCTC 8385), Staphylococcus aureus (ATCC 25923), Lyesria moncytogenes (ATCC 19116) and Streptococcus pneumonia (ATCC 63). Results also showed considerable antimicrobial activity of garlic and tazma honey. MIC of mixture of garlic and tazma honey at 6.25% against total test bacteria was 88.9%. MIC of mixture of garlic and tazma honey at 6.25% against Gram positive and negative were 100% and 83.33%, respectively. The bactericidal activities of garlic, tazma honey, and mixture of garlic and tazma honey against all pathogenic bacteria at 6.25% concentration were 66.6%, 55.6% and 55.6%, respectively. Conclusions This finding strongly supports the claim of the local community to use the combination of tazma honey and garlic for the treatment of different pathogenic bacterial infections. Therefore, garlic in combination with tazma honey can serve as an alternative natural antimicrobial drug for the treatment of pathogenic bacterial infections. Further in vivo study is recommended to come up with a comprehensive conclusion.

737-742

Authors: Ponmurugan Karuppiah, Muhammed Mustaffa

Objective To investigate different Musa sp. leave extracts of hexane, ethyl acetate and methanol were evaluated for antibacterial activity against multi-drug resistant pathogens causing nosocomial infection by agar well diffusion method and also antioxidant activities. Methods The four different Musa species leaves were extracted with hexane, ethyl acetate and methanol. Antibacterial susceptibility test, minimum inhibitory concentration and minimum inhibitory bacterial concentration were determined by agar well diffusion method. Total phenolic content and in vitro antioxidant activity was determined. Results All the Musa sp. extracts showed moderate antibacterial activities expect Musa paradisiaca with the inhibition zone ranging from 8.0 to 18.6 mm. Among four species ethyl acetate extracts of Musa paradisiaca showed highest activity against tested pathogens particularly E. coli, P. aeruginosa and Citrobacter sp. The minimum inhibitory concentrations were within the value of 15.63- 250 µg/mL and minimum bactericidal concentrations were ranging from 31.25- 250 µg/mL. Antioxidant activity of Musa acuminate exhibited maximum activity among other three Musa species. Conclusions The present study concluded that among the different Musa species, Musa paradisiaca displayed efficient antibacterial activity followed by Musa acuminata against multi-drug resistant nosocomial infection causing pathogens. Further, an extensive study is needed to identify the bioactive compounds, mode of action and toxic effect in vivo of Musa sp.

748-750

Authors: Mahdavi Seif Ali, Raeesi Ahmad, Faraji Leyla, Youssefi Mohammad Reza, Rahimi Mohammad Taghi

A case of malaria infection in a 42-year-old woman in rural area of Mahmodabad, Mazandaran Province, North Iran was reported and discussed elaborately. She was complaining about recurrent fevers, sweating, headache and myalgia in back. After her first admission to hospital due to misdiagnose she did not receive proper treatment and the patient suffered from clinical manifestations again. Eventually in the second admission to another hospital, after a precise examination on her thick and thin blood smear the agent of disease was recognized appropriately as Plasmodium vivax and treated accordingly.

595-603

Authors: Shereen Farouk Mossallam, Eglal Ibrahim Amer, Iman Fathy Abou-El-Naga

Objective To investigate the effect of rotifer internalization into snail tissue on the development of schistosomes. Methods Susceptible laboratory-bred Biomphalaria alexandrina (B. alexandrina) snails were exposed to lab-maintained rotifers; Philodina spp., two weeks before and after being infected with Schistosoma mansoni (S. mansoni) miracidia. The consequent histopathological impact on snail tissues and cercarial biology were investigated before and after emergence from snails. Results Contamination of B. alexandrina snails with philodina, two weeks before miracidial exposure, was found to hinder the preliminary development of S. mansoni cercariae inside the snail tissues. Furthermore, when snails were contaminated with rotifers two weeks post miracidial exposure; growth of already established cercariae was found to be retarded. The consequent influence of internalized rotifers within the snail tissue was clearly reflected on cercarial emergence, activity and infectivity along the four weeks duration of shedding. In the present study, comparison of snail histopathological findings and altered cercarial biology observed between the experimental and control groups indicated that the rotifers may have affected the levels of snail's energy reservoirs, which eventually was found to have had an adverse impact on reproduction, growth and survival of the parasite within the snail host, coupled with its performance outside the snail. Conclusions In future biological control strategies of schistosomiasis, ritifers should be considered as a parasitic scourge of humanity.

611-616

Authors: Imad Abdelhamid El-Haci, Fawzia Atik Bekkara, Wissame Mazari, Fayçal Hassani, Mohamed Amine Didi

Objective To investigate the antioxidant and the antibacterial activities of crude extract from aerial part of Polygonum maritimum L. (Polygonaceae) (P. maritimum) and to find new actives biomolecules. Methods The whole plant was collected from the Rechgoune coast (West of Algeria), and methanolic crude extract of aerial parts of P. maritimum (PMCE) was prepared. The extract was tested against different bacterial strain and tested for his ability to neutralize free radical (DPPH) and to scavenge the H2O2. Results PMCE had a very high content of total phenol, which was (352.49±18.03) mg/g dry weight, expressed as gallic acid equivalent. PMCE exhibited excellent antioxidant activity, as measured using DPPH and H2O2 scavenging assays. It also showed a high antibacterial activity against gram-positive bacterial strains: Bacillus cereus, Bacillus subtilis and Staphylococcus aureus with an highest MIC of 120 µg/mL. Conclusions The antioxidant and antibacterial activity of the PMCE is probably due to phenolic compounds present in the extract. The contemporary presence of antioxidant and antibacterial activities in the PMCE suggests that this plant may be a source of bioactive substances with multifaceted activity.

623-627

Authors: Fatma Alhakmani, Sokindra Kumar, Shah Alam Khan

Objective To evaluate and compare the antioxidant potential and anti-inflammatory activity of ethanolic extract of flowers of Moringa oleifera (M. oleifera) grown in Oman. Methods Flowers of M. oleifera were collected in the month of December 2012 and identified by a botanist. Alcoholic extract of the dry pulverized flowers of M. oleifera were obtained by cold maceration method. The ethanolic flower extract was subjected to preliminary phytochemical screening as the reported methods. Folin-Ciocalteu reagent was used to estimate total phenolic content. DPPH was used to determine in-vitro antioxidant activity and anti-inflammatory activity of flowers was investigated by protein denaturation method. Results Phytochemical analysis of extract showed presence of major classes of phytochemicals such as tannins, alkaloids, flavonoids, cardiac glycosides etc. M. oleifera flowers were found to contain 19.31 mg/g of gallic acid equivalent of total phenolics in dry extract but exhibited moderate antioxidant activity. The anti-inflammatory activity of plant extract was significant and comparable with the standard drug diclofenac sodium. Conclusions The results of our study suggest that flowers of M. oleifera possess potent anti-inflammatory activity and are also a good source of natural antioxidants. Further study is needed to identify the chemical compounds responsible for their anti-inflammatory activity.

634-638

Authors: Prawez Alam

Objective To develop and validate a simple, accurate HPTLC method for the analysis of 8-gingerol and to determine the quantity of 8-gingerol in Zingiber officinale extract and ginger-containing dietary supplements, teas and commercial creams. Methods The analysis was performed on 10×20 cm aluminium-backed plates coated with 0.2 mm layers of silica gel 60 F254 (E-Merck, Germany) with n-hexane: ethyl acetate 60: 40 (v/v) as mobile phase. Camag TLC Scanner III was used for the UV densitometric scanning at 569. Results This system was found to give a compact spot of 8-gingerol at retention factor (Rf) value of (0.39±0.04) and linearity was found in the ranges 50-500 ng/spot (r2=0.9987). Limit of detection (12.76 ng/spot), limit of quantification (26.32 ng/spot), accuracy (less than 2 %) and recovery (ranging from 98.22-99.20) were found satisfactory. Conclusions The HPTLC method developed for quantification of 8-gingerol was found to be simple, accurate, reproducible, sensitive and is applicable to the analysis of 8-gingerol in Zingiber officinale extract and ginger-containing dietary supplements, teas and commercial creams.

644-649

Authors: Meltem Demirel Kars, Ufuk Gündüz, Kamil Üney, Ahmet Levent Baş

Objective To investigate the molecular effects of Nerium oleander leaf distillate on paclitaxel and vincristine resistant (MCF-7/Pac and MCF-7/Vinc) cells and sensitive (MCF-7/S) cell lines. Methods Nerium oleander (N. oleander) leaf extract was obtained by hydrodistillation method. The toxicological effects of N. oleander distillate, previously suggested as medicinal food supplement, on drug resistant cells were evaluated by XTT tests. MDR modulation potential of the plant material was evaluated by flow cytometry and fluorescent microscopy. Paclitaxel and vincristine were applied to the sublines in combination with N. oleander distillate. Results Fractional inhibitory indices show that N. oleander distillate did not increase the antiproliferative effects of anticancer drugs. N. oleander treatment in to MCF-7/Pac and MCF-7/Vinc did not inhibit P-gp activity and MDR1 gene expression level. Conclusions As a result it may be suggested that although N. oleander distillate has some medicinal effects as food supplement it may not be suitable as an MDR modulator for drug resistant breast cancer cells.

657-662

Authors: Eshetu Molla, Mirutse Giday, Berhanu Erko

Objective To assess the molluscicidal and cercariacidal activities of aqueous extracts of Balanites aegyptiaca (B. aegyptiaca) against Ethiopian Biomphalaria pfeifferi (B. pfeifferi), Lymnaea natalensis (L. natalensis) and Schistosoma mansoni (S. mansoni) cercariae. Methods Extracts of seeds, endocarp, mesocarp, and fruit of B. aegyptiaca were tested for their activities against adult B. pfeifferi and L. natalensis. The cercariacidal activity of the seeds of the plant was also evaluated against S. mansoni. Bioassays were carried out following the methods recommended by WHO. Snail mortalities were compared between each plant part and snail species, and LC50 and LC90 values for the plant parts tested were computed. The cercariacidal activity of the plant was assessed by exposing the mice to the cercariae pre-exposed to aqueous extract of B. aegyptiaca seeds. Results For the molluscicidal activities of seeds, endocarp, mesocarp and whole fruit, the LC50 values against B. pfeifferi were 56.32, 77.53, 65.51 and 66.63 mg/L, respectively, while the respective LC90 values were 77.70, 120.04, 89.50 and 97.55 mg/L. Similarly, the LC50 values for the seeds, endocarp, mesocarp and whole fruit against L. natalensis were 80.33, 92.61, 83.52 and 87.84 mg/L, respectively, while the respective LC90 values were 102.30, 138.21, 115.42 and 127.69 mg/L. B. pfeifferi were found to be more susceptible to B. aegyptiaca than L. natalensis. S. mansoni cercariae exposed to 15 mg/L of extract of seeds were incapable of infecting mice. The mean egg load of tissue was reduced in mice infected with the cercariae exposed to 5 and 10 mg/L of the extract. Conclusions The aqueous extracts of different parts of B. aegyptiaca exhibited reasonable molluscicidal activity against B. pfeifferi and L. natalensis, as well as cercariacidal activity against S. mansoni cercariae. However, comprehensive laboratory evaluation is recommended prior to field tests of the plant parts since their impact on other aquatic biota is not known.

668-672

Authors: Sathyanarayana Bhat, Kumaraswamy Udupa

Origin of ancient Indian toxicology can be dated back to vedic literature. Toxins of both animate and inanimate world were very well understood during the era. Rig and Atharva vedic texts describe such details. After classifying such toxins, Charaka Samhitha, thebasic literature of Indian Medicine used gold and ghee as panaceas to counter act them. Ayurveda considers toxicology as one among the eight specialized branches of medical wisdom. Unfortunately, the available literature on this is very limited. Moreover, they have been discussed briefly in Charaka and Sushrutha Samhitha. Mangarasa I, a Jain scholar who lived on the foothills of the Western Ghats, in Southern India in 1350 A.D., felt this vacuum and composed an independent, elaborate Kannada text on toxicology. His less known text Khagendra Mani Darpana (KMD) is the first ever documented complete text on toxicology in the world. Medieval Indian wisdom on plant and animal diversities are very well reflected in this unique toxicological text. Centuries past to Linnean era, KMD gives vivid descriptions on zoological and botanical diversities of the time. This astonishing fact is an evidence of our ancestor's curiosities about the nature around them. A critical overview of the bio-diversity described in KMD text is discussed in this paper.

515-522

Authors: Koosha Paydary, Parisa Khaghani, Sahra Emamzadeh-Fard, SeyedAhmad SeyedAlinaghi, Kazem Baesi

After its identification in 1980s, HIV has infected more than 30 million people worldwide. In the era of highly active anti-retroviral therapy, anti-retroviral drug resistance results from insufficient anti-retroviral pressure, which may lead to treatment failure. Preliminary studies support the idea that anti-retroviral drug resistance has evolved largely as a result of low-adherence of patients to therapy and extensive use of anti-retroviral drugs in the developed world; however, a highly heterogeneous horde of viral quasi-species are currently circulating in developing nations. Thus, the prioritizing of strategies adopted in such two worlds should be quite different considering the varying anti-retroviral drug resistance prevalence. In this article, we explore differences in anti-retroviral drug resistance patterns between developed and developing countries, as they represent two distinct ecological niches of HIV from an evolutionary standpoint.

532-535

Authors: Bruno Peixoto, Isabel Kalei

Objective To characterize the neurocognitive sequelae of cerebral malaria (CM) in an adult sample of the city of Benguela, Angola. Methods A neuropsychological assessment was carried out in 22 subjects with prior history of CM ranging from 6 to 12 months after the infection. The obtained results were compared to a control group with no previous history of cerebral malaria. The study was conducted in Benguela Central Hospital, Angola in 2011. Results CM group obtained lower results on the two last trials of a verbal learning task and on an abstract reasoning test. Conclusions CM is associated to a slower verbal learning rate and to difficulties in the ability to discriminate and perceive relations between new elements.

542-548

Authors: Hussein Farhan, Hassan Rammal, Akram Hijazi, Ahmad Daher, Mohamad Reda, Hussein Annan, Ali Chokr, Ali Bassal, Bassam Badran

Objective To determine the chemical composition, total phenolic and total flavonoid contents of the crude extracts from leaves and stems of a Lebanese plant Euphorbia macroclada schyzoceras (E. macroclada), and to evaluate their antioxidant potential using DPPH, H2O2, and chelating of ferrous ions tests. Methods Quantification of the total phenolic and total flavonoid contents of the crude extracts from leaves and stems and the antioxidant activities were evaluated using spectrophotometric analyses. The chemical composition has been estimated using different techniques such as IR, LC/MS and NMR. Results Ethanolic extract from leaves of E. macroclada was better than aqueous extract and showed higher content in total phenolic and total flavonoid than found in the stems. On the other hand, using DPPH and H2O2 tests, this extract from leaves showed higher antioxidant capacity than aqueous extract. However, using the chelating of ferrous ions test, the antioxidant activity of the aqueous extract of both stems and leaves was stronger than that of ethanolic once. The chemical composition of the whole plant showed the presence of some aromatic compounds and fatty acids. Conclusions Both ethanolic and water extracts from both parts of this plant are effective and have good antioxidant power. So, this plant can be used in the prevention of a number of diseases related to oxidative stress.

554-556

Authors: Marslin Gregory, B. Divya, Revina Ann Mary, M. M. Hipolith Viji, V. K. Kalaichelvan, V. Palanivel

Objective To evaluate the anti-ulcer activity and acute toxicity of Ficus religiosa (F. religiosa) leaf ethanolic extract in animal models. Methods Anti-ulcer activity of F. religiosa ethanolic extract (250 and 500 mg/kg body weight) was studied on stress induced ulcer animal models. Ranitidine was used as standard. The anti-ulcer activity of F. religiosa was evaluated with the help of ulcer area and histopatholgical examination. Preliminary phyto-chemical screening and acute toxicity studies of F. religiosa also carried out. Results Results showed that the extract treatments prevented ulcer area and gastric secretion in a dose-dependent manner. Administration of 2 000 mg/kg extract did not show any acute toxicity in albino mice. Preliminary phytochemical analysis identified the presence of flavonoids in the ethanolic extract of F. religiosa. Conclusions The extract is non-toxic even at relatively high concentrations. The anti-ulcer activity is probably due to the presence of flavanoids.

563-568

Authors: Sherif W. Mansour, Sibghatullah Sangi, Sree Harsha, Mueen A. Khaleel, A. R. N. Ibrahim

Objective To clarify the modulatory effects of daily consumption of pomegranate extract (PE), olive oil (OO) and Nagilla sativa oil (NSO) on antioxidant activity, sperm quality and pituitary-testicular axis of adult male wistar rats. Methods Thirty-two adult male Wistar rats were divided into four equal groups, eight rats each. Using rat gastric tubes, 1.0 mL distilled water, 1.0 mL PE, 0.4 mL NSO and 0.4 mL OO were orally administered daily for 6 weeks in the first, second, third and fourth groups, respectively. Reproductive organs, body weight, sperm criteria, testosterone, FSH, LH, inhibin-B, lipid peroxidation, and antioxidant enzyme activities were investigated. At the end of the study protocol, analyses occurred at the same time. Data were analysed by ANOVA test and P<0.05 was considered to be a significant value. Results In all studied groups, malondialdehyde level was significantly decreased accompanied with an increases in glutathione peroxidase and glutathione. Rats treated with PE showed an increase in catalase activities accompanied with an increase in sperm concentration which was also observed in NSO group. In PE treated group, sperm motility was also increased accompanied with decreased abnormal sperm rate. NSO, OO and PE treated groups shows an insignificant effect on testosterone, inhibin-B, FSH and LH in comparison with control group. Conclusions These results show that administration of PE, NSO and OO could modify sperm characteristics and antioxidant activity of adult male wistar rats.

573-579

Authors: Saber Abdelruhman Sakr, Mohamed El-said Zoil, Samraa Samy El-shafey

Objective To evaluate the ameliorative role of grapefruit juice on the cytogenetic and testicular damage induced by the antiarrythmic drug amiodarone in albino rats. Methods Animals were divided into four groups. Group I was considered as control. Group II was given grapefruit juice at a dose level of 27 mL/kg body weight. Group III was orally administered amiodarone (18 mg/kg body weight) daily for 5 weeks. Animals were sacrificed after 5 weeks of treatment. Bone marrow was collected from the femurs for analysis of chromosomal aberrations and mitotic indices. Testes were removed and stained with H&E for histological examination. Sperms were collected from epidedymis for detection of sperm head abnormalities. Comet assay was used to detect DNA damage. Results Amiodarone treatment caused a significant increase in the percentage of chromosomal aberrations, decreased the mitotic index and increased DNA damage. The testis showed many histopathological alterations, inhibition of spermatogenesis and morphometric changes. The number of sperm head abnormalities was increased. Treating animals with amiodarone and grapefruit juice caused a reduction in chromosomal aberrations, mitotic index, DNA damage and testicular alterations caused by amiodarone. Conclusions The results of this study indicated that grapefruit juice ameliorates the cytotoxicty and testicular alterations induced by amiodarone in albino rats and this is may be due to the potent antioxidant effects of its components.

584-585

Authors: Viroj Wiwanitkit

no

587-588

Authors: Samia Meliani, Bouabdellah Benallou, Miloud Halbouche, Zohra Haddouche

Objective To enhance effectiveness of reproduction management in Arabian mares, factors influencing the time of foaling were investigated in this study. Methods Data were collected at the National Haras of Tiaret in Algeria from 2003 to 2010. The foaling time of 255 Arabian pure bred mares, aged from 3 to 20 years were used for this study. Results A total of 78.07% of foaling happens between 7 pm and 6 am. Conclusions The influence of the month of foaling and the sex of the foal, on the time of foaling was statically significant.