Asian Pacific Journal of Tropical Biomedicine

Issue 8,2012 Table of Contents

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  • 1  Synergistic anticancer effect of the extracts from Polyalthia evecta caused apoptosis in human hepatoma (HepG2) cells
    Sasipawan Machana Natthida Weerapreeyakul Sahapat Barusrux Kanjana Thumanu Waraporn Tanthanuch
    [Abstract](21) [HTML](0) [PDF 398.25 K](124)
    Objective: To evaluate the anticancer activity of the extract fraction of Polyalthia evecta (P. evecta) (Pierre) Finet & Gagnep and the synergistic anticancer effect of the extracts from P. evecta by using the ATR/FT-IR spectroscopy. Methods: The 50% ethanol-water crude leaf extract of P. evecta (EW-L) was prepared and was further fractionated to isolate various fractions. The anticancer activity was investigated from cytotoxicity against HepG2 using a neutral red assay and apoptosis induction by evaluation of nuclei morphological changes after DAPI staining. Synergistic anticancer effects of the extracts from P. evecta were performed using the ATR/FT-IR spectroscopy. Results: The result showed that the EW-L showed higher cytotoxicity and apoptosis induction in HepG2 cells than its fractionated extracts. The hexane extract exhibited higher cytotoxicity and apoptosis induction than the water extracts, but less than the EW-L. The combined water and hexane extracts apparently increased cytotoxicity and apoptosis induction. The %apoptotic cells induced by the extract mixture were increased about 2-fold compared to the single hexane extract. Conclusions: The polar extract fraction is necessary for the anticancer activity of the non-polar extract fraction. The ATR/FT-IR spectra illustrates the physical interaction among the constituents in the extract mixture and reveals the presence of polyphenolic constituents in the EW-L, which might play a role for the synergistic anticancer effect.
    2  Antibacterial effect of Allium sativum cloves and Zingiber officinale rhizomes against multiple-drug resistant clinical pathogens
    Ponmurugan Karuppiah Shyamkumar Rajaram
    [Abstract](22) [HTML](0) [PDF 284.50 K](135)
    Objective: To evaluate the antibacterial properties of Allium sativum (garlic) cloves and Zingiber officinale (ginger) rhizomes against multi-drug resistant clinical pathogens causing nosocomial infection. Methods: The cloves of garlic and rhizomes of ginger were extracted with 95% (v/v) ethanol. The ethanolic extracts were subjected to antibacterial sensitivity test against clinical pathogens. Results: Anti-bacterial potentials of the extracts of two crude garlic cloves and ginger rhizomes were tested against five gram negative and two gram positive multi-drug resistant bacteria isolates. All the bacterial isolates were susceptible to crude extracts of both plants extracts. Except Enterobacter sp. and Klebsiella sp., all other isolates were susceptible when subjected to ethanolic extracts of garlic and ginger. The highest inhibition zone was observed with garlic (19.45 mm) against Pseudomonas aeruginosa (P. aeruginosa). The minimal inhibitory concentration was as low as 67.00 μg/mL against P. aeruginosa. Conclusions: Natural spices of garlic and ginger possess effective anti-bacterial activity against multi-drug clinical pathogens and can be used for prevention of drug resistant microbial diseases and further evaluation is necessary.
    3  Repellent properties of Cardiospermum halicacabum Linn. (Family: Sapindaceae) plant leaf extracts against three important vector mosquitoes
    M Govindarajan R Sivakumar
    [Abstract](19) [HTML](0) [PDF 285.55 K](128)
    Objective: To determine repellent activity of hexane, ethyl acetate, benzene, chloroform and methanol extract of Cardiospermum halicacabum (C. halicacabum) against Culex quinquefasciatus (Cx. quinquefasciatus), Aedes aegypti (Ae. aegypti) and Anopheles stephensi (An. stephensi). Methods: Evaluation was carried out in a net cage (45 cm×30 cm×25 cm) containing 100 blood starved female mosquitoes of three mosquito species and were assayed in the laboratory condition by using the protocol of WHO 2005; The plant leaf crude extracts of C. halicacabum was applied at 1.0, 2.5, and 5.0 mg/cm2 separately in the exposed area of the fore arm. Only ethanol served as control. Results: In this observation, the plant crude extracts gave protection against mosquito bites without any allergic reaction to the test person, and also, the repellent activity was dependent on the strength of the plant extracts. The tested plant crude extracts had exerted promising repellent against all the three mosquitoes. Conclusions: From the results it can be concluded the crude extract of C. halicacabum was potential for controlling Cx. quinquefasciatus, Ae. aegypti and An. stephensi mosquitoes.
    4  Aqueous extracts of microalgae exhibit antioxidant and anticancer activities
    Sanaa MM Shanab Soha SM Mostafa Emad A Shalaby Ghada I Mahmoud
    [Abstract](19) [HTML](0) [PDF 346.15 K](122)
    Objective: To investigate the antioxidant and anticancer activities of aqueous extracts of nine microalgal species. Methods: Variable percentages of major secondary metabolites (total phenolic content, terpenoids and alkaloids) as well as phycobiliprotein pigments (phycocyanin, allophycocyanin and phycoerythrin) in the aqueous algal extracts were recorded. Antioxidant activity of the algal extracts was performed using 2, 2 diphenyl-1-picrylhydrazyl (DPPH) test and 2,2'- azino-bis (ethylbenzthiazoline-6-sulfonic acid (ABTS.+) radical cation assay. Anticancer efficiency of the algal water extracts was investigated against Ehrlich Ascites Carcinoma cell (EACC) and Human hepatocellular cancer cell line (HepG2). Results: Antioxidant activity of the algal extracts was performed using DPPH test and ABTS.+ radical cation assays which revealed 30.1-72.4% and 32.0-75.9% respectively. Anticancer efficiency of the algal water extracts was investigated against Ehrlich Ascites Carcinoma Cell (EACC) and Human Hepatocellular cancer cell line (HepG2) with an activity ranged 87.25% and 89.4% respectively. Culturing the promising cyanobacteria species; Nostoc muscorum and Oscillatoria sp. under nitrogen stress conditions (increasing and decreasing nitrate content of the normal BG11 medium, 1.5 g/L), increased nitrate concentration (3, 6 and 9 g/L) led to a remarkable increase in phycobilin pigments followed by an increase in both antioxidant and anticancer activities in both cyanobacterial species. While the decreased nitrate concentration (0.75, 0.37 and 0.0 g/L) induced an obvious decrease in phycobilin pigments with complete absence of allophycocyanin in case of Oscillatoria sp. Conclusions: Nitrogen starvation (0.00 g/L nitrate) induced an increase and comparable antioxidant and anticancer activities to those cultured in the highest nitrate content.
    5  In vitro scolicidal effect of Satureja khuzistanica (Jamzad) essential oil
    Mohammad Moazeni Mohammad Jamal Saharkhiz Ali Akbar Hoseini Amir Mootabi Alavi
    [Abstract](20) [HTML](0) [PDF 280.46 K](130)
    Objective: To investigate the scolicidal effect of the Satureja khuzistanica (S. khuzistanica) essential oil from aerial parts of this herbal plant. Methods: The essential oil was obtained by hydrodistillation method. Gas chromatography (GC) and gas chromatography mass spectrometry (GC-MS) were employed to determine the chemical composition of the essential oil. Protoscolices were collected aseptically from sheep livers containing hydatid cyst. Protoscolices were exposed to various concentrations of the oil (3, 5 and 10 mg/mL) for 10, 20, 30, and 60 min. Viability of protoscolices was confirmed by 0.1% eosin staining. Results: A total of 19 compounds representing 97.6% of the total oil, were identified. Carvacrol (94.9%) was found to be the major essential oil constituent. Scolicidal activity of S. khuzistanica essential oil at concentration of 3 mg/mL was 28.58, 32.71, 37.20 and 42.02%, respectively. This essential oil at concentration of 5 mg/ mL killed 51.33, 66.68, 81.12, and 100% of protoscolices after 10, 20, 30 and 60 min, respectively. One hundred scolicidal effect was observed with S. khuzistanica essential oil at the concentration of 10 mg/mL after 10 min (comparing with 7.19% for control group). Conclusions: The essential oil of S. khuzistanica is rich in carvacrol and may be used as a natural scolicidal agent.
    6  Genetic analysis of maturity and flowering characteristics in maize (Zea mays L.)
    Hassan Sher Muhammad Iqbal Kiramat Khan Muhammad yasir Hameed-ur-Rahman
    [Abstract](23) [HTML](0) [PDF 276.34 K](127)
    Objective: To elucidate the pattern of inheritance and determine the relative magnitude of various genetic effects for maturity and flowering attributes in subtropical maize. Methods: Four white grain maize inbred lines from flint group of corn, two with late maturity and two with early maturity, were used. These contrasting inbred lines were crossed to form four crosses. Six generations (P1, P2, F1, F2, BC1, and BC2) were developed for each individual cross. These were evaluated in triplicate trial for two consecutive years. Results: Both dominance gene action and epistatic interaction played major role in governing inheritance of days to pollen shedding, 50% silking, anthesis silking interval and maturity. Conclusions: Preponderance of dominance gene action for these traits indicated their usefulness in hybrid programs of subtropical maize.
    7  Evaluation of anti-inflammatory potential of leaf extracts of Skimmia anquetilia
    Vijender Kumar Zulfiqar Ali Bhat Dinesh Kumar NA Khan IA Chashoo
    [Abstract](19) [HTML](0) [PDF 262.05 K](169)
    Objective: To evaluate anti-inflammatory potential of leaf extract of Skimmia anquetilia by in-vitro and in-vivo anti-inflammatory models. Methods: Acute toxicity study was carried out to determine the toxicity level of different extract using acute toxic class method as described in Organization of Economic Co-operation and Development Guidelines No.423. Carrageenan (1% w/w) was administered and inflammation was induced in rat paw. The leaf extracts of Skimmia anquetilia were evaluated for anti-inflammatory activity by in-vitro human red blood cell (HRBC) membrane stabilization method and in-vivo carrangeenan-induced rat paw edema method. Results: The in-vitro membrane stabilizing test showed petroleum ether (PE), chloroform (CE), ethyl acetate (EE), methanol (ME) and aqueous extracts (AE) showed 49.44%, 59.39%, 60.15%, 68.40% and 52.18 % protection, respectively as compared to control groups. The in-vivo results of CE, EE and ME showed 58.20%, 60.17% and 67.53% inhibition of inflammation after 6h administration of test drugs in albino rats. The potency of the leaf extracts of Skimmia anquetilia were compared with standard diclofenac (10 mg/kg) which showed 74.18% protection in in-vitro HRBC membrane stabilization test and 71.64% inhibition in in-vivo carrangeenan-induced rat paw edema model. The ME showed a dose dependent significant (P< 0.01) anti-inflammatory activity in human red blood cell membrane stabilization test and reduction of edema in carrageenan induced rat paw edema. Conclusions: The present investigation has confirmed the anti-inflammatory activity of Skimmia anquetilia due to presence of bioactive phytoconstitutes for the first time and provide the pharmacological evidence in favor of traditional claim of Skimmia anquetilia as an anti-inflammatory agent.
    8  Toxicity of cadmium and lead on tropical midge larvae, Chironomus kiiensis Tokunaga and Chironomus javanus Kieffer (Diptera: Chironomidae)
    Warrin Ebau Che Salmah Md Rawi Zubir Din Salman Abdo Al-Shami
    [Abstract](18) [HTML](0) [PDF 267.20 K](145)
    Objective: To investigate the acute toxicity of cadmium and lead on larvae of two tropical Chironomid species, Chironomus kiiensis (C. kiiensis) Tokunaga and Chironomus javanus (C. javanus) Kieffer. Methods: Different larval instars (first-fourth) were exposed using a static non-replacement testing procedures to various concentrations of cadmium and lead. Results: In general, younger larvae (first and second instars) of both species were more sensitive to both metals than older larvae (third and forth instars). The toxic effects of the metals on C. kiiensis and C. javanus were influenced by the age of the larvae (first to fourth instars), types of metals (cadmium or lead) and duration of larval exposure (24, 48, 72 and 96 h) to the metals. Conclusions: Cadmium was more toxic to the chironomids than lead and C. javanus was significantly more sensitive to both metals than C. kiiensis (P<0.05).
    9  Anti-dermatophytic activity of marine sponge, Sigmadocia carnosa (Dendy) on clinically isolated fungi
    NB Dhayanithi TT Ajith Kumar M Kalaiselvam T Balasubramanian N Sivakumar
    [Abstract](23) [HTML](0) [PDF 292.34 K](129)
    Objective: To screen the anti-fungal effects and find out the active metabolites from sponge, Sigmadocia carnosa (S. carnosa) against four dermatophytic fungi. Methods: The methanol, ethyl acetate and acetone extract of marine sponge, S. carnosa was examined against Trichophyton mentagrophytes (T. mentagrophytes), Trichophyton rubrum (T. rubrum), Epidermophyton floccosum (E. floccosum) and Microsporum gypseum (M. gypseum) and qualitative analysed to find out the active molecules. Results: The methanol extract of sponge was expressed significant activity than ethyl acetate and acetone. The minimum inhibitory concentration (MIC) of methanol extract of sponge that resulted in complete growth inhibition of T. mentagrophytes, T. rubrum, E. floccosum and M. gypseum were found to 125, 250, 250 and 250 μg/mL respectively. But, 100 % inhibition of fungal spore germination was observed in T. mentagrophytes at 500 μg/mL concentration followed by T. rubrum, E. floccosum and M. gypseum at 1 000 μg/mL concentration. Other two extracts showed weak anti spore germination activity against the tested dermatophytic fungi. Methanol extracts showed presence of terpenoids, steroids, alkaloids, saponins and glycosides. Conclusion: Based on the literature, this is the first study which has conducted to inhibit the growth and spore germination of dermatophytic fungi with S. carnosa. Further research also needs to purify and characterize the secondary metabolites from the sponge, S. carnosa for the valuable source of novel substances for future drug discovery.
    10  Science behind cisplatin-induced nephrotoxicity in humans: A clinical study
    Arunkumar PA Viswanatha GL Radheshyam N Mukund H Belliyappa MS
    [Abstract](16) [HTML](0) [PDF 266.80 K](128)
    Objective: To investigate the relationship between serum electrolyte changes and cisplatin induced nephrotoxicity. Methods: We collected data from 18 patients undergoing cisplatin chemotherapy including serum electrolytes, creatinine, blood urea nitrogen (BUN) and urine potassium, sodium and pH levels before and after the cisplatin chemotherapy. All the patients had cancer and were treated with 40-50 mg/day cisplatin. Renal injury was assessed by measuring serum electrolytes, creatinine, BUN levels and urine potassium, sodium and pH levels. Results: The five cycles of cisplatin based chemotherapy resulted in hypomagnesia (P=0.029), hypocalcaemia (P=0.001*), hypophosphatemia (P=0.003*), hypokalemia (P=0.001*) and increased serum creatinine (P=0.001*) and BUN (P=0.292*) levels. In urine analysis, decrease in potassium (P=0.024*) was found, except potassium there was no significant changes in sodium and urine pH. Conclusions: The present study demonstrates that, acute nephrotoxicity was observed in patients with different types of cancers undergoing cisplatin based chemotherapy due to electrolyte disturbances, when no corrective measures were initiated.
    11  Alleviating effects of melatonin on oxidative changes in the testes and pituitary glands evoked by subacute chlorpyrifos administration in Wistar rats
    Angela J Umosen Suleiman F Ambali Joseph O Ayo Bisala Mohammed Chidiebere Uchendu
    [Abstract](23) [HTML](0) [PDF 346.86 K](120)
    Objective: To evaluate the alleviating effects of melatonin on oxidative changes in the testes and pituitary gland induced by subacute chlopyrifos (CPF) exposure in rats. Methods: Forty adult male Wistar rats divided into 4 groups of 10 animals were used for the study. Group Ⅰ received soya oil (2 mL/kg) while group Ⅱ was administered with melatonin (0.5 mg/kg). Group Ⅲ was administered CPF only (8.5 mg/kg ~ 1/10th of the LD50) while group Ⅳ was pretreated with melatonin (0.5 mg/kg) and then exposed to CPF (8.5 mg/kg), 10 min later. The regimens were administered by gavage once daily for a period of 28 d. At the end of the exposure period, the rats were sacrificed and the testicular tissues and pituitary glands were evaluated for the malonaldehyde (MDA) concentration and activities of superoxide dismutase (SOD) and catalase (CAT). Results: CPF increased MDA concentrations and reduced the activities of SOD and CAT in the testes and pituitary gland. Melatonin pretreatment reduced the testicular and pituitary MDA concentrations and improves the SOD and CAT activities. Conclusions: the study showed that subacute CPF-induced oxidative stress in the testes and pituitary glands were alleviated by melatonin due to its antioxidant property.
    12  Analysis of chemical composition and bioactive property evaluation of Indian propolis
    R Thirugnanasampandan Sayana Beena Raveendran R Jayakumar
    [Abstract](22) [HTML](0) [PDF 270.79 K](128)
    Objective: To analyze the chemical composition and to evaluate the bioactive potential of hydroalocoholic extract of propolis. Methods: Ethanol extract of propolis was analyzed by GC-MS, HPTLC and HPLC methods and in vitro antioxidant, anticholinesterase and cytotoxicity assay were performed. Results: GC-MS analysis revealed the presence of fatty acids, alcohols, and quercetin. Quercetin was identified and quantified by HPTLC and HPLC methods. Dose dependent DPPH and hydroxyl radical scavenging activity of hydroalcoholic extract of propolis was calculated as 16.20 and 34.33 μg/mL respectively. Inhibition of lipid peroxidation was significant and the IC50 value was calculated as 55.56μg/mL. Anticholinesterase activity was less observed. The cytotoxic activity against both breast (MCF-7) and lung cancer (A543) cell lines were significant and the IC50 value was calculated as 10 and 13 μg/mL respectively. Conclusions: These findings showed that bioactive compounds present in propolis will alleviate many diseases and can be used for better human health.
    13  Bacterial and fungal endophthalmitis in Upper Egypt: related species and risk factors
    AA Gharamah AM Moharram MA Ismail AK AL-Hussaini
    [Abstract](24) [HTML](0) [PDF 337.63 K](127)
    Objective: To study risk factors, contributing factors of bacterial and fungal endophthalmitis in Upper Egypt, test the isolated species sensitive to some therapeutic agents, and to investigate the air-borne bacteria and fungi in opthalmology operating rooms. Methods: Thirty one cases of endophthalmitis were clinically diagnosed and microbiologically studied. Indoor air-borne bacteria and fungi inside four air-conditioned operating rooms in the Ophthalmology Department at Assiut University Hospitals were also investigated. The isolated microbes from endophthalmitis cases were tested for their ability to produce some extracellular enzymes including protease, lipase, urease, phosphatase and catalase. Also the ability of 5 fungal isolates from endophthalmitis origin to produce mycotoxins and their sensitivity to some therapeutic agents were studied. Results: Results showed that bacteria and fungi were responsihle for infection in 10 and 6 cases of endophthalmitis, respectively and only 2 cases produced a mixture of bacteria and fungi. Trauma was the most prevalent risk factor of endophthalmitis where 58.1% of the 31 cases were due to trauma. In ophthalmology operating rooms, different bacterial and fungal species were isolated. 8 bacterial and 5 fungal isolates showed their ability to produce enzymes while only 3 fungal isolates were able to produce mycotoxins. Terbinafine showed the highest effect against most isolates in vitro. Conclusions: The ability of bacterial and fungal isolates to produce extracellular enzymes and mycotoxins may be aid in the invasion and destruction of eye tissues. Microbial contamination of operating rooms with air-borne bacteria and fungi in the present work may be a source of postoperative endophthalmitis.
    14  A review on medicinal importance, pharmacological activity and bioanalytical aspects of beta-carboline alkaloid ‘‘Harmine’’
    K Patel M Gadewar R Tripathi SK Prasad Dinesh Kumar Patel
    [Abstract](20) [HTML](0) [PDF 284.16 K](138)
    Harmine, a beta-carboline alkaloid, is widely distributed in the plants, marine creatures, insects, mammalians as well as in human tissues and body fluids. Harmine was originally isolated from seeds of Peganum harmal in 1847 having a core indole structure and a pyridine ring. Harmine has various types of pharmacological activities such as antimicrobial, antifungal, antitumor, cytotoxic, antiplasmodial, antioxidaant, antimutagenic, antigenotoxic and hallucinogenic properties. It acts on gamma-aminobutyric acid type A and monoamine oxidase A or B receptor, enhances insulin sensitivity and also produces vasorelaxant effect. Harmine prevents bone loss by suppressing osteoclastogenesis. The current review gives an overview on pharmacological activity and analytical techniques of harmine, which may be useful for researcheres to explore the hidden potential of harmine and and will also help in developing new drugs for the treatment of various diseases.
    15  A review on ethnobotany, phytochemistry and pharmacology of Fumaria indica (Fumitory)
    Prakash Chandra Gupta Nisha Sharma Ch V Rao
    [Abstract](19) [HTML](0) [PDF 253.28 K](136)
    Fumaria indica (Hausskn.) Pugsley (Fumariaceae), known as “Fumitory”, is an annual herb found as a common weed all over the plains of India and Pakistan. The whole plant is widely used in traditional and folkloric systems of medicine. In traditional systems of medicine, the plant is reputed for its anthelmintic, diuretic, diaphoretic, laxative, cholagogue, stomachic and sedative activities and is used to purify blood and in liver obstruction in ethnopharmacology. The whole plant is ascribed to possess medicinal virtues in Ayurvedic and Unani systems of medicine and is also used in preparation of important Ayurvedic medicinal preparations and polyherbal liver formulations. The review reveals that phytochemical constituents of wide range have been separated from the plants and it possesses important pharmacological activities like smooth muscle relaxant, spasmogenic and spasmolytic, analgesic, anti-inflammatory, neuropharmacological and antibacterial activities. The separation of hepatoprotective and antifungal constituents from this plant was also reported newly. This review highlights the traditional, ethnobotanical, phytochemical, pharmacological information available on Fumaria indica, which might be helpful for scientists and researchers to find out new chemical entities responsible for its claimed traditional uses.
    16  Metastatic apocrine sweat gland adenocarcinoma in a terrier dog
    Akhtardanesh Baharak Kheirandish Reza Dabiri Shahriar Azari Omid Vosoogh Daruoosh Askari Nasrin
    [Abstract](24) [HTML](0) [PDF 283.05 K](129)
    This report describes the clinical and pathological aspects of an apocrine sweat gland carcinoma with distant metastasis in an aged dog. A 7-year-old male terrier dog was referred to small animal hospital of Shahid Bahonar University of Kerman with a 5.5×3.5 centimeter pedunculated mass on its head near left auricular region which had been progressively growing since three months ago. The radiography showed no local and distant metastasis. Surgical excision and histological evaluation was done. Histologically, the mass was composed of epithelial cells arranged in glandular and solid patterns. The morphologic findings suggested either a primary or metastatic apocrine-gland carcinoma. Immunohistochemically, the tumor cells were intensely positive for cytokeratin 7 and 20 and negative for S100 protein. On the basis of histopathological and clinical findings, the tumor was diagnosed as a malignant apocrine gland tumor, arising from apocrine sweat glands of the skin. Local tumor recurrence with anorexia and weight loss was reported by the owner nine month later. Severe submandibular and prescapular lymphadenomegaly was noted in clinical examination. Several large pulmonary nodules were noted in chest radiographs resembling mediastinal lymph node metastasis. Second surgery and chemotherapy was rejected by the owner due to grave prognosis of the patient. The animal was died 45 days later due to respiratory complications. Tumors of apocrine sweat glands are relatively uncommon in dogs whereas apocrine gland adenocarcinoma with distant metastasis is extremely rare.

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