Asian Pacific Journal of Tropical Biomedicine

Issue 11,2013 Table of Contents

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  • 1  Assessment of antidiabetic activity and acute toxicity of leaf extracts from Physalis peruviana L. in guinea-pig
    Félicien Mushagalusa Kasali Justin Ntokamunda Kadima Pius Tshimankinda Mpiana Koto-te-Nyiwa Ngbolua Damien Sha-Tshibey Tshibangu
    2013(11):841-846. DOI: 10.1016/S2221-1691(13)60166-5
    [Abstract](20) [HTML](0) [PDF 755.46 K](79)
    Abstract:
    Objective: To verify the antidiabetic activity of leaf extracts from Physalis peruviana L. popularly used in the Eastern part of the Democratic Republic of the Congo and to point out the possible toxicity. Method: Aqueous decoctions prepared from dried leaves powder were administrated to guinea pigs at the dose range of 100 mg/kg to 3.2 g/kg of body weight. The hypoglycemic activity was evaluated by glucose tolerance test, loading animals with glucose 4 g/kg and measuring blood glucose concentrations at various times. The effect was compared to the control and glibenclamide as antidiabetic reference drug. Acute toxicity was evaluated by recording mortality rate, changes on blood biomarkers and damage caused to vital organs. Results: At a dose of 100 mg/kg, the aqueous extract induced a significant reduction of peak concentration at 30 min after glucose loading as compared with control or reference (P<0.05). At doses greater than 400 mg, some alterations on blood, kidney and liver markers were observed. Upper 800 mg/kg, mortality was observed with LD50 estimated at about 1 280 mg/kg. At the autopsy, vital organs were in haemorrhage and swelling state. Conclusion: The crude aqueous extracts from the leaves of Physalis peruviana L. present hypoglycemic activity in animal model, but at high doses the plant may cause severe intoxication.
    2  Combination of low-concentration of novel phytoestrogen (8,9)-furanyl-pterocarpan-3-ol from Pachyrhizus erosus attenuated tamoxifen-associated growth inhibition on breast cancer T47D cells
    Arief Nurrochmad Endang Lukitaningsih Ameilinda Monikawati Dita Brenna Septhea Edy Meiyanto
    2013(11):847-852. DOI: 10.1016/S2221-1691(13)60167-7
    [Abstract](23) [HTML](0) [PDF 1013.38 K](75)
    Abstract:
    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.
    3  Evaluation of larvicidal activity of Pongamia pinnata extracts against three mosquito vectors
    Guna Ranjan Kolli Balakrishnan Vijayan Raja Sundararajan
    2013(11):853-858. DOI: 10.1016/S2221-1691(13)60168-9
    [Abstract](22) [HTML](0) [PDF 733.52 K](73)
    Abstract:
    Objective: To evaluate the mosquito larvicidal activity of Pongamia pinnata (P. pinnata) extracts against three mosquito vectors. Methods: The methanol and hydroalcohol extracts of bark part of P. pinnata L were tested against fourth instar larvae of Culex quinquefasciatus, Aedes aegypti and Anopheles stephensi. The mortality was observed 24 h and 48 h after treatment, data was subjected to probit analysis to determine lethal concentration (LC50 and LC90) to kill 50 and 90 percent of treated larvae of tested species. Results: The larval mortality was found in both methanol and hydroalcohol extracts of P. pinnata against Culex quinquefasciatus, Aedes aegypti and Anopheles stephensi with LC50 values of 84.8, 118.2 and 151.7 ppm; 97.7, 128.3 and 513 ppm. The highest larval mortality was found in methanol extract of P. pinnata when comparable to the hydroalcohol extract. Conclusions: These results suggest that both methanol and hyrdoalcohol extracts have the potential to be used as an ideal ecofriendly approach for the control of disease vectors. This could lead to isolation of novel natural larvicidal compounds.
    4  In vitro analysis on bactericidal screening and antioxidant potentiality of leaf and root extracts of Thottea siliquosa (Lam.) Ding Hou. An ethnobotanical plant
    Saiba Abdul Wahab Nusaiba Kumaraswamy Murugan
    2013(11):859-865. DOI: 10.1016/S2221-1691(13)60169-0
    [Abstract](14) [HTML](0) [PDF 755.90 K](71)
    Abstract:
    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.
    5  In vitro hypoglycemic effects of Albizzia lebbeck and Mucuna pruriens
    Mangesh Bhutkar Satish Bhise
    2013(11):866-870. DOI: 10.1016/S2221-1691(13)60170-7
    [Abstract](6) [HTML](0) [PDF 394.82 K](73)
    Abstract:
    Objective: To verify the antidiabetic potential of stem bark of Albizzia lebbeck (A. lebbeck) and seeds of Mucuna pruriens (M. pruriens) using various in vitro techniques. Methods: The plant extracts were studied for their effects on glucose adsorption, diffusion amylolysis kinetics and glucose transport across yeast cells. Results: Both the plant extracts adsorbed glucose and the adsorption of glucose increased remarkably with an increase in glucose concentration. No significant (P≤0.05) differences were observed between the adsorption capacities of A. lebbeck and M. pruriens. In amylolysis kinetic experimental model the rate of glucose diffusion was found to increase with time from 30 to 180 min, and both the plant extracts demonstrated significant inhibitory effects on movement of glucose into external solution across dialysis membrane as compared to control. The retardation of glucose diffusion by A. lebbeck extract was significantly higher (P≤0.05) than M. pruriens. These effects were reflected with higher glucose dialysis retardation index values for A. lebbeck than M. pruriens. The plant extracts also promoted glucose uptake by yeast cells. The rate of uptake of glucose into yeast cells was linear in all the 5 glucose concentrations used in the study. M. pruriens extract exhibited significantly higher (P≤0.05) activity than the extract of A. lebbeck at all concentrations. Conclusions: The results verified the antidiabetic potential of A. lebbeck and M. pruriens. The hypoglycemic effect exhibited by the extracts is mediated by increasing glucose adsorption, decreasing glucose diffusion rate and at the cellular level by promoting glucose transport across the cell membrane as revealed by simple in vitro model of yeast cells.
    6  Phytochemical investigation and in vitro antioxidant activity of an indigenous medicinal plant Alpinia nigra B.L. Burtt
    Suprava Sahoo Goutam Ghosh Debajyoti Das Sanghamitra Nayak
    2013(11):871-876. DOI: 10.1016/S2221-1691(13)60171-9
    [Abstract](30) [HTML](0) [PDF 735.56 K](76)
    Abstract:
    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.
    7  Hepatoprotective effect of leaves of Erythroxylum monogynum Roxb. on paracetamol induced toxicity
    Sabeena Hussain Syed Ajay Gajanan Namdeo
    2013(11):877-881. DOI: 10.1016/S2221-1691(13)60172-0
    [Abstract](24) [HTML](0) [PDF 873.20 K](70)
    Abstract:
    Objective: To investigate the hepatoprotective activity of methanolic extract of leaves of Erythroxylum monogynum (E. monogynum) on paracetamol induced toxicity. Methods: Methanolic extract of leaves of E. monogynum was given in doses of 100 mg/kg, 200 mg/ kg and 400 mg/kg for 7 d and toxicity was induced by paracetamol (2 mg/kg) on Day 8. Silymarin (50 mg/kg) was used as reference standard. After 24 h of toxicity induction blood samples were collected from retro-orbital plexsus and analyzed for serum parameters like serum glutamic pyruvic transaminase, serum glutamic oxaloacetate transaminse, alkaline phosphatase and total bilirubin. Livers isolated were studied for histopathological changes. Results: Phytochemical analysis of methanolic extract of E. monogynum leaves showed the presence of carbohydrates, flavonoids, phenols and saponins. Prior administration of this extract restored the elevated levels serum markers as compared to toxic group which is also confirmed by the histopathological changes observed. Conclusions: The present study showed that methanolic extract of leaves of E. monogynum possess hepatoprotective action against paracetamol induced hepatotoxicity.
    8  Ethnomedicinal plants used in the treatment of skin diseases in Hyderabad Karnataka region, Karnataka, India
    Shivakumar Singh Policepatel Vidyasagar Gunagambhire Manikrao
    2013(11):882-886. DOI: 10.1016/S2221-1691(13)60173-2
    [Abstract](15) [HTML](0) [PDF 731.21 K](69)
    Abstract:
    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.
    9  Phytochemical analysis and in vitro antioxidant acitivity of hydroalcoholic seed extract of Nymphaea nouchali Burm. f.
    Mabel Parimala Francis Gricilda Shoba
    2013(11):887-895. DOI: 10.1016/S2221-1691(13)60174-4
    [Abstract](33) [HTML](0) [PDF 773.93 K](73)
    Abstract:
    Objective: To evaluate the phytochemical constituents and the antioxidant activity of hydroalcoholic extract of Nymphaea nouchali seed locally prescribed as a diet for diabetes mellitus. Methods: The antioxidant and free radical scavenging activity of hydroalcoholic extract of the plant was assessed against 1,1 diphenyl-2-picryl hydrazyl (DPPH), nitric oxide and lipid peroxidation using standard protocols. Total phenolics, flavonoids and tannins were also determined. Results: Phytochemical analysis revealed the presence of phenols, flavones, tannins, protein, reducing sugars, glycosides, saponins, alkaloids and steroids. The activities of plant extract against DPPH, nitric oxide and lipid peroxidation was concentration dependent with IC50 value of 42.82, 23.58 and 54.65 µg/mL respectively. The total antioxidant capacity was high with 577.73 mg vitamin E/g of the extract and showed a moderately high vitamin C content of 197.22 mg/g. The total tannin content of hydroalcoholic seed extract was high (195.84 GE/g), followed by phenolics (179.56 GE/g) and flavonoids (23.55 QE/g). Conclusion: Our findings provide evidence that the crude extract of Nymphaea nouchali is a potential source of natural antioxidants and this justifies its use in folkloric medicine.
    10  Evaluation of cytotoxic and anti-tumor activity of partially purified serine protease isolate from the Indian earthworm Pheretima posthuma
    Mahendra Kumar Verma Francies Xavier Yogendra Kumar Verma Kota Sobha
    2013(11):896-901. DOI: 10.1016/S2221-1691(13)60175-6
    [Abstract](14) [HTML](0) [PDF 754.13 K](72)
    Abstract:
    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.
    11  Evaluation of genotoxicity of Trois through Ames and in vitro chromosomal aberration tests
    Manu Chaudhary Anurag Payasi
    2013(11):902-906. DOI: 10.1016/S2221-1691(13)60176-8
    [Abstract](19) [HTML](0) [PDF 683.28 K](71)
    Abstract:
    Objective: To investigate the mutagenic potential of Trois using the bacterial reverse mutation assay (Ames test) and in vitro chromosomal aberration test. Methods: The ability of Trois to induce reverse mutations was evaluated in Salmonella typhimurium (TA 98, TA100, TA1535 and TA1537) and Escherichia coli (WP2 uvrA) with and without metabolic activation system (S9 mix) at the dose range of 313 to 5000 µg/plate. Chromosomal aberrations were evaluated in Chinese hamster lung (CHL) cell line at the dose levels of 15, 7.5, 3.7, 1.9 and 0.9 mg/mL in the absence and presence of S9 mix. Results: There were no increases in the number of revertant colonies at any concentrations of Trois used in the study with and without S9 mix in all tester strains. Trois did not produce any structural aberration in CHL cells in the presence or absence of S9 mix. Conclusions: Results of this study suggest that Trois is non-mutagenic.
    12  Comparison of lipid profile in different grades of non-alcoholic fatty liver disease diagnosed on ultrasound
    Dhumal Uttareshvar Mahaling Madole Mahesh Basavaraj Aher Jagdish Bika
    2013(11):907-912. DOI: 10.1016/S2221-1691(13)60177-X
    [Abstract](23) [HTML](0) [PDF 977.99 K](73)
    Abstract:
    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.
    13  Clinical and hematological study of canine Ehrlichiosis with other hemoprotozoan parasites in Kolkata, West Bengal, India
    Mousam Das Sabyasachi Konar
    2013(11):913-915. DOI: 10.1016/S2221-1691(13)60178-1
    [Abstract](26) [HTML](0) [PDF 697.80 K](69)
    Abstract:
    Objective: To observe other hemoprotozoan diseases with canine ehrlichiosis and to evaluate the clinical and hematological aspects of dogs naturally infected with ehrlichiosis with other hemoprotozoan diseases. Methods: Blood was collected for hematological value and Giemsa stained blood smear was made for diagnosis of Ehrlichia sp. and other hemoprotozoan parasites from naturally infected dogs. Case history was taken from the owner and clinical signs and symptoms were noted. Results: A total of 47 cases of ehrlichiosis in dogs were reported with babesiosis (8.51%) and hepatozoonosis (6.38%) hemoprotozoan diseases. Ehrlichia canis, Ehrlichia ewingii, Brucella canis, Babesia gibsoni and Hepatozoon canis were observed under oil immersion lense of microscope in Giemsa stained peripheral blood smears. Marked anaemia and neutrophilic leukocytosis were observed. Conclusions: The results of this study stated that clinical and haematological changes occurred in canine ehrlichiosis with babesiosis and hepatozoonosis due to parasitemia. In mixed infection, the disease more severe, and also it depended on immunity of animals. Babesia gibsoni and Hepatozoon canis with Ehrlichia sp. were first reported from West Bengal state of India by this study.
    14  Cryptosporidiosis as threatening health problem: A review
    Nurul Fariza Rossle Baha Latif
    2013(11):916-924. DOI: 10.1016/S2221-1691(13)60179-3
    [Abstract](17) [HTML](0) [PDF 856.33 K](74)
    Abstract:
    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.

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