Asian Pacific Journal of Tropical Biomedicine

85-91

Authors: Netiya Karaket, Nuchada Maneejantra, Patoomratana Tuchinda, Jirapha Kunapin, Choowong Auesukaree, Kanyaratt Supaibulwatana

bjective: To explore the effect of gibberellic acid (GA3) and its inhibitor paclobutrazol (PBZ) on chemical composition and their pharmacological effects on Andrographis paniculata (Burm. f.) Wall. ex Nees, and to clarify action mode of andrographolide. Methods: The chemical composition was extracted by sequential extraction with hexane, dichloromethane, ethyl acetate and methanol, respectively. Andrographolide and its derivatives were evaluated by HPLC. Moreover, the metabolic profiling was analyzed by GC-MS. Inhibitory effect of crude extracts was tested against Staphylococcus aureus using agar well diffusion method. Mode of action was tested against mutant yeast by spotting assay. Andrographolide were tested for their mode of action against eukaryotes. Results: Among different solvents, dichloromethane gave the highest yield of crude (3.58% DW), with the highest andrographolide content (8.3 mg/g DW). The effect of plant hormone (10 mg/L GA3 or PBZ) on phytochemical variations and bioactivity of Andrographis paniculata was demonstrated. It was found that PBZ promoted sesquiterpene compounds about 3.5 times over than GA3 treatment. But inhibitory effect of extracts against Staphylococcus aureus was highest in GA3 treated plants; andrographolide and 14-deoxy-11,12-didehydroandrographolide contents were significantly higher than those of water or PBZ. It was found that there were 11 strains involving in ergosterol biosynthesis, V-ATPase activity and homeostasis, and superoxide detoxification process. In this regard, andrographolide might cause the damage on the lipid bilayer of yeast cell and plasma membrane by interfering ergosterol biosynthesis. Conclusions: It is found that GA3 promotes andrographolide and 14-deoxy-11,12-didehydroandrographolide content while PBZ promotes sesquiterpene content. Andrographolide might cause the damage on the lipid bilayer of yeast cell and plasma membrane by interfering ergosterol biosynthesis. It might also affect mitochondria electron transport chain, leading to the occurrence of ROS, which can further harm cell organelles. However, the library screening is the first step to investigate mode of action of andrographolide.

98-105

Authors: Valentine Chi Mbatchou, Rita Akosua Dickson, Isaac Kingsley Amponsah, Abraham Yeboah Mensah, Solomon Habtemariam

Objective: To explore the potential insecticidal, ovipositor deterrent and antifeedant effects of ethyl acetate extract of the seeds of Senna tora (Syn. Cassia tora) against cowpea weevil (Callosobruchus maculatus). Methods: The activities were evaluated using standard protocols. In these bioassays, the cowpea seeds were used directly as an insect feed. The activity of the extract and isolated compounds were tested at concentrations of 100, 200 and 300 μg/mL and compared to neem oil and cinnamaldehyde (as standard positive controls). Phytochemical analysis of the ethyl acetate extract was done through a number of chromatographic techniques and the structures of the isolated compounds were established through comprehensive spectroscopic analysis including 2D-NMR and ESI-MS studies. Results: Fractionation of the active ethyl acetate extract resulted in the isolation of one known anthraquinone, aurantio-obtusin (1) and a novel compound that was named as cassiatorin (2). Compounds 1 and 2 showed comparable insect antifeedant properties with the positive controls while their insecticidal and ovipositor deterrent effects were far superior to the standard controls. Conclusions: It is thus concluded that Senna tora extracts and the isolated compounds (1 and 2) may be employed in the postharvest management of stored cowpea seeds and as other crop protectants.

113-119

Authors: Zihan Rahman Khan, Nargis Sultana Chowdhury, Suriya Sharmin, Md. Hossain Sohrab

Nymphoides is an aquatic genus consisting about 50 species, of which few were accepted as traditional medicinal plants in Asia. The literature review revealed that Nymphoides species are widely used in Ayurvedic medicine as a popular drug, i.e. Tagara. They are also utilized by the traditional local healers of different Asian countries to treat various diseases, like convulsion, jaundice, fever, headache, etc. According to the in vivo and in vitro pharmacological studies, Nymphoides species have been claimed to possess major biological activities like anticonvulsant, antioxidant, hepatoprotective, cytotoxic, antitumor, etc. Biochemical profiling of different aquatic plants of this genus revealed the presence of some important phytochemicals as polyphenolic component, flavonoids, triterpenes, carbohydrates, glycosides, etc. Several valuable bioactive compounds including ephedrine, coumarin, secoiridoid glucosides, methyl quercetin, ferulic acid, foliamenthoic acid, etc. were also known to be isolated and identified from different Nymphoides species. The aim of this review is to analyze the published report based on the medicinal values of different Asian Nymphoides species, to provide the updated information about the ethnomedicinal, pharmacological as well as the phytochemical properties for the first time.

130-136

Authors: Alfred Maroyi

Diospyros lycioides Desf. (D. lycioides) is traditionally used as herbal medicine against various human and animal ailments in tropical Africa. The present paper reviewed information on botany, medicinal uses, phytochemistry and pharmacological activities of D. lycioides. This review was compiled using scientific literature from electronic search engine such as PubMed, Scopus, ScienceDirect, Springerlink, BioMed Central, Scielo, Medline and Science domain. Additional literatures were obtained from book chapters, books, dissertations, websites and other scientific publications. D. lycioides is used as traditional medicine in 50% of the countries where the species is native in tropical Africa. This study recorded 22 medicinal uses of D. lycioides which included abdominal pains, infertility in women, sexually transmitted infections, and used as chewing sticks (or mouthwash), toothbrushes and ethnoveterinary medicine. D. lycioides extracts demonstrated anti-adhesive, anti-inflammatory, antimetastatic, antioxidant, antifungal, antiproliferative, mutagenicity and antibacterial activities. Future research should focus on the pharmacological properties, phytochemistry, clinical trials and pharmacokinetics of D. lycioides which will enhance the therapeutic potential of the species.

565-570

Authors: Zohreh Khaki, Seyedeh Parastoo Yasini, Seyedeh Missagh Jalali

Objective: To determine the biochemical and acute phase proteins changes in sheep experimentally infected with Anaplasma ovis (A. ovis). Methods: One Iranian sheep naturally infected with A. ovis (parasitemia 0.02%) but with no other blood parasites based on blood smear and polymerase chain reaction methods was selected as donor, and it was splenectomized to induce high level of parasitemia. Then, three weeks after splenectomy when parasitemia was 6%, donor’s blood was intravenously administered to each recipient animal. Five 5-6 months old Iranian male sheep without any blood parasites were selected as recipient animals. The percent of parasites, packed cell volume, serum biochemical parameters (urea, creatinine, bilirubin, aspartate aminotransferase activity, cholesterol, total protein, albumin, globulin, Fe), acute phase proteins (haptoglobin, total iron binding capacity, fibrinogen), were evaluated in sheep before and after being experimentally infected with A. ovis (until day 38). In addition, body weights of sheep were measured on days 0, 20 and 38. Results: In recipient sheep, microscopic examination of erythrocytes revealed a significant rise of parasitemia on days 12 and 15. The lowest level of packed cell volume in sheep was seen on day 15 post infection. A significant rise existed in mean urea and bilirubin (total, direct and indirect) on days 15 and 20. The increase of indirect bilirubin level was higher than direct bilirubin. Furthermore, serum Fe significantly increased on days 20 and 23. The mean total protein concentration significantly increased on day 38. A significant increase was found in the serum globulin concentration from days 20 and 27 to 38. Maximum values of haptoglobin were observed on days 27 and 30. Moreover, aspartate aminotransferase activity (from days 20-30) and cholesterol concentration (on day 20) significantly decreased. However, no significant changes were found in other parameters. Conclusions: Experimental ovine anaplasmosis caused by A. ovis could be associated with some changes in measured parameters, which presumably could be helpful for evaluation on staging of disease.

580-585

Authors: Nittaya Chansiw, Narisara Paradee, Kamonnaree Chotinantakul, Somdet Srichairattanakool

Objective: To investigate anti-hemolytic, antibacterial and anti-cancer activities of leaf and stem extracts from Polygonum odoratum. Methods: Leaves and stems of Polygonum odoratum were extracted using methanol and their anti-hemolytic activity was assessed using 2, 2′-Azobis (2-methylpropionamidine) dihydrochloride which is known to generate free radical damage on cell membranes of red blood cells. This damage, represented by hemolysis, was measured using spectrophotometry. Antibacterial activity was tested by using a broth microdilution method to find minimal inhibitory concentrations against eight bacterial strains. Anti-cancer activity of the extracts was evaluated against a human promyelocytic leukemic cell line (HL- 60) by using MTT assay for cell viability and flow cytometry for apoptosis induction and cell cycle analysis. Results: Both leaf and stem extracts have anti-hemolytic activity. The results showed a significantly increased percentage of inhibition in a concentration-dependent manner. Interestingly, the leaf extract showed anti-hemolytic activity to a greater extent than the stem extract. Antibacterial activity of the extracts, as indicated by their minimal inhibitory concentration, using 12.5, 50, 25, 25 μg/mL, was measured against Staphylococcus epidermidis, Enterococcus faecium, Enterococcus faecalis and Staphylococcus aureus. The leaf extracts also exhibited anti-cancer activity, demonstrated by significantly decreased cell viability of human promyelocytic cells (HL-60), with an IC50 of (350.00±1.85) μg/mL for 48 h and (38.00±0.92) μg/mL for 72 h. Additionally, HL-60 became apoptotic and accumulated in G1–phase after 48 hours of treatment. Conclusions: The extracts of Polygonum odoratum exhibit potential anti-hemolytic activity. They also have antibacterial activity by inhibiting growth of Gram-positive bacteria. The leaf extract shows anti-cancer activity against HL-60 to a greater extent than the stem extract, causing decreased viability, increased G1-phase accumulation and apoptosis induction.

593-597

Authors: Bamidele S Ajilore, Abdulfatah A Adesokan

Objective: To investigate antidiabetic effects of Tetracarpidium conophorum seed (TECOSE) against biomarkers of diabetes-induced nephropathy in rats. Methods: Powdered seed (500 g) of TECOSE was extracted with 5 L of 100% methanol for 72 h followed by concentration of filtrate. Diabetes was induced in rats with 75 mg/kg body weight intraperitoneal streptozotocin. The rats were randomly divided into five groups (n=5 in each group) namely group A- normal control, group B- diabetic control, groups C, D and E were diabetic rats treated with 500 mg/kg body weight TECOSE orally, 7 mg/kg body weight metformin orally and subcutaneous 0.3 IU/kg body weight HumulinR, respectively. All rats were treated once daily for 2 weeks. Blood samples and urine were collected for biochemical estimations. Kidney was harvested for histomorphological studies. Results: TECOSE (500 mg/kg body weight) significantly (P<0.05) reduced blood sugar levels as effective as metformin and insulin. The plant extract also significantly (P<0.05) reduced levels of serum urea, creatinine and uric acid, proteinuria, ketonuria, hematuria and glycosuria while it significantly (P<0.05) increased glomerular filtration rate. Histomorphological study of the kidney of untreated diabetic rats showed features suggestive of glomerulosclerosis and tubular necrosis while that of treatments with TECOSE extract, metformin and insulin showed near normal histoarchitectures. Conclusions: Streptozotocin at 75 mg/kg body weight induces diabetic nephropathy in rats and TECOSE possesses potentials to prevent diabetic renal damage.

604-608

Authors: Jarinyaporn Naowaboot, Pritsana Piyabhan, Pholawat Tingpej, Narongsuk Munkong, Wason Parklak, Patchareewan Pannangpetch

Objective: To evaluate the insulin sensitivity action of ferulic acid (FA) in skeletal muscle and hypothalamus of high-fat diet (HFD)-induced obese mice. Methods: Obese mouse model was induced by HFD (45 kcal% lard fat) for 16 weeks. After 8 weeks of HFD feeding, these obese mice were orally treated with FA at doses of 25 and 50 mg/kg/day for 8 weeks. At the end of all treatments, the epididymal fat, pancreas, skeletal muscle and hypothalamus were removed for biochemical parameter and protein expression examinations. Results: FA treatment significantly decreased leptin level in fat tissue and insulin level in pancreas (P < 0.05). Interestingly, obese mice treated with FA increased the protein expressions of insulin receptor substrate-1, phosphatidylinositol 3-kinase, and phosphorylated-protein kinase B in both muscle and brain (P < 0.05). The phosphorylations of adenosine monophosphate-activated protein kinase and acetyl-CoA carboxylase in muscle, and leptin receptor protein in hypothalamus were also increased (P < 0.05). The pancreatic islets histology showed smaller size in obese mice treated with FA compared to untreated obese mice. Conclusions: These findings indicate the beneficial effect of FA in improving insulin resistance in HFD-induced obese mice. These effects are probably mediated via modulating the insulin receptor substrate/phosphatidylinositol 3-kinase/protein kinase B or adenosine monophosphate-activated protein kinase pathways.

519-526

Authors: Teeranai Ittiudomrak, Songchan Puthong, Tanapat Palaga, Sittiruk Roytrakul, Chanpen Chanchao

Objective: To find new compounds in order to overcome the mainstay of metastatic breast cancer due to the adverse side effects from, and increasing resistance to, current chemotherapeutic agents. Methods: α-Mangostin and apigenin were reported in comparison to doxorubicin, a chemotherapeutic drug. Ductal carcinoma (BT474) cell line and non-tumorigenic epithelial tissue from mammary gland (MCF-10A) were used. Cell viability assessment was calculated by the standard 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide method. Cell morphology was investigated by light microscopy. By flow cytometry analysis, programmed cell death was observed using annexin V and propidium iodide staining while cell-cycle arrest was observed using propidium iodide staining. Change in transcriptional expression was evaluated by real-time quantitative reverse transcription PCR. Results: In 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay, the result revealed α-mangostin and apigenin were more cytotoxic to BT474 cells. Longer exposure times to α-mangostin and apigenin caused more floating cells and a lower density of adhered cells with more vacuoles present in the colonies in BT474 only. α-Mangostin and apigenin caused necrosis in BT474 cells in a 24 h exposure, but a small amount of early apoptotic cells could also be detected at 24, 48 and 72 h exposure, whereas doxorubicin caused early apoptosis to BT474 cells at 24 h. Transcript expression and activity analysis supported caspase-3 was involved in the death of BT474 cells treated by all compounds. Moreover, α-mangostin and apigenin arrested the cell-cycle at the G1-phase, but at the G2/M-phase by doxorubicin. All three compounds induced a change in transcript expression levels of inflammation-associated, proto-oncogene, autophagy-associated and apoptosis-associated genes. Conclusions: α-Mangostin and apigenin are worth investigating as potential new sources of chemotherapeutic agents for breast cancer treatment.

533-539

Authors: Muhammad Eka Prastya, Rika I Astuti, Irmanida Batubara, Aris T Wahyudi

Objective: To analyze potential activation of oxidative stress tolerance systems by SAB E-41 bacterial extract in promoting the life span of yeast Schizosaccharomyces pombe. Methods: In vitro analysis was done to assess antioxidant activity of SAB E-41 bacterial extract. Antiaging property of the particular extract was then assayed through spot test and chronological life span assays. Furthermore, sty1 mitogen-activated protein kinase, pap1 transcriptional factor of oxidative stress response and its downstream genes, ctt1 were evaluated via real time PCR. The protein level of ctt1 was then observed via Western Blot analysis. In addition, accumulation of reactive oxygen species and mitochondrial activity were conducted to understand the effect of SAB E-41 upon oxidative stress response systems in vivo. Results: The IC50 values of corresponding extract for antioxidant (DPPH; ABTS) and antiglycation were 402.40, 358.13 and 683.55 μg/mL, respectively. In addition, SAB E-41 extract (750 μg/mL) exhibited antiaging properties, which could be attributed to significant up-regulation of oxidative stress response genes, sty1, pap1 and ctt1. Interestingly, SAB E-41 extract could enhance stress tolerance phenotype of Schizosaccharomyces pombe against H2O2-induced oxidative stress. These results were supported by increasing mitochondrial activity and reactive oxygen species intracellular levels. Conclusions: SAB E-41 extract could promote yeast life span likely via up-regulation of oxidative stress responses in yeast. Our results suggest that adaptive response via up-regulation of oxidative stress transcriptional factors, and its downstream gene, ctt1, as well as mitochondrial activity contributes in combating oxidative stress thus promoting yeast life span.

548-553

Authors: Hilal Irmak Sapmaz, Azibe Yıldız, Alaadin Polat, Nigar Vardı, Evren Köse, Kevser Tanbek, Songül Çuğlan

Objective: To explore the effects of Nigella sativa oil (NSO) on the histopathological and biochemical changes that inhaled formaldehyde (FA) induces on the testicular tissue of rats. Methods: Thirty three adult male rats were separated into five groups as follows: C, the control group; 4FA group which received FA for 4 weeks; 13FA group which was given FA for 13 weeks; 4FA+NSO group which was administered FA plus NSO for 4 weeks; 13FA+NSO group which was treated with FA plus NSO for 13 weeks. FA was administered through inhalation for 8 h 5 days a week at a dose of 5 ppm in a special glass cage, and NSO was administered orally 1 mL/kg once daily. Rats were decapitated at the end of the experiment and testicular tissue specimens were harvested for histopathologic and biochemical assessment. Results: Compared to the C group, reduction was observed in the number of intact tubules and in the mean germinative epithelium thickness of the FA groups. Significant increase was observed in the number of intact tubules with the long-term (13 weeks) administration of NSO together with FA. Reduced glutathione peroxidase activity was found and oxidative stress index values were measured higher in the 4FA and 13FA groups versus the C group (P<0.05). Moreover, total antioxidant status levels decreased only in the 4FA group (P<0.05) while only the 13FA group significantly increased malondialdehyde levels and reduced catalase activities in comparison with the C group. In the 13FA+NSO group, malondialdehyde levels decreased however glutathione peroxidase and catalase activities increased compared to the 13FA group. Differences measured in total antioxidant status levels were found to be statistically significant only between the 4FA and the 4FA+NSO groups. Conclusions: NSO as an antioxidant should be used for a longer term to achieve protective efficacy both histopathologically and biochemically in the testicular tissue.

471-477

Authors: Bhagavathi Sundaram Sivamaruthi, Periyanaina Kesika, Kumaresan Subasankari, Chaiyavat Chaiyasut

Anthocyanins (ACN) are water-soluble pigments, belonging to flavonoids, and are present in almost all fruits, and vegetables at varying concentration. About 635 ACN were distinguished based on the position and number of methoxyl and hydroxyl moieties in the basic structure of ACN. Pelargonidin, cyanidin, delphinidin, malvidin, peonidin, and petunidin are extensively studied anthocyanidins. The absorption, bioavailability, metabolism, pharmacokinetics, molecular mechanism, and analytical techniques of several phytochemicals were described. The biological benefits (antidiabetic, anti-neuro-disorder, anti-cardiovascular diseases, antigastrointestinal diseases, and disorders) of flavonoids and ACN have been reported. Several in vitro, and in vivo reports demonstrated that ACN-rich plant extracts ameliorate the diabetes-associated consequences by reducing the glucose absorption, ROS production, oxidative stress, glomerular angiogenesis, lipid synthesis, and FoxO1 and adipose triglyceride lipase expressions, and improve the insulin secretion, insulin sensitivity, glucose tolerance, glucose uptake, glucose consumption, antioxidant activity. The literature search was made in Scopus, Google Scholar, PubMed using the keywords “anthocyanin” and “diabetes”. The documents were carefully checked for the relevance to the current manuscript and the selection was made without any chronological restriction. The present manuscript summarizes the updated reports on antihyperglycemic properties of ACN.

485-489

Authors: JW Lee, W Wu, SY Lim

Objective: To evaluate the antioxidant activity of extracts and fractions from Stachys sieboldii Miq., and to examine its effect on the cellular reactive oxygen species (ROS) and glutathione (GSH) production and genomic DNA oxidation in HT-1080 cells. Methods: The ROS generation induced by H2O2 was measured by the dichlorofluorescein-diacetate assay. GSH levels were measured using a fluorescent method with mBBr. Genomic DNA oxidative damage was measured with levels of oxidative DNA induced by the reaction of ferritin with H2O2. Results: The n-hexane, 85% aqueous methanol and n-butanol fractions (0.05 mg/mL concentrations) inhibited H2O2-induced ROS generation by 63%, 35% and 45%, respectively. GSH levels were significantly increased in both acetone+methylene chloride and methanol extracts (P<0.05). Supplementation of cells with n-hexane significantly increased GSH levels at concentrations of 0.05 mg/mL (P<0.05). Both the acetone+methylene chloride and methanol extracts, as well as all fractions significantly inhibited oxidative DNA damage (P<0.05). Conclusions: These results indicate that cellular oxidation was inhibited by the n-hexane fraction and this fraction may contain valuable active compounds.

500-512

Authors: Kolambe Rajappa Chethana, Balappa Somappa Sasidhar, Mahadeva Naika, Rangappa Sangappa Keri

Objective: To screen plant extract fractions and elucidate the components present in Caesalpinia crista (C. crista) leaves for cholinergic and anti-amyloidogenic activities for the treatment of Alzheimer's diseases. Methods: This work has been carried out to study the action of C. crista extracts from nonpolar to polar solvents toward inhibition of oxidative stress, cholinergic and amyloidosis. The antioxidant activity was studied using DPPH total antioxidant assay; cholinergic assay by Ellman's method and anti-amyloidogenic assay by thioflavin-T fluorescence and transmission electron microscopy. Results: The quantification of polyphenols was carried out following C. crista methanolic extract (CCMeOH) HPLC fingerprinting, along with LC-MS and elucidated by MS LAMPS database. GC-MS of CCMeOH was screened for potential moieties. In vitro experimental results showed that the CCMeOH was potential extract that exhibited active inhibition of antioxidant property, cholinergic enzymes acetylcholinesterase and butyrylcholinesterase. For anti-amyloidogenic evaluations, among all the extracts, the CCMeOH was found to have the potential toward inhibiting the oligomers, fibrillation of Aβ42 with good defibrillation of amyloid cascading properties. Conclusions: These results are also supported by the presence of polyphenols as the active ingredients. Multi-potent target drug therapy is a promising option in treating the Alzheimer's diseases. Methanolic extract of C. crista shows potential activity against cholinergic enzymes, Aβ42 aggregation with antioxidant activity.

1-6

Authors: Jinho Park, Du-Gyeong Han, SuHee Kim, Jeong-Byoung Chae, Joon-Seok Chae, Do-Hyeon Yu, Kyoung-Seong Choi

Objective: To investigate the prevalence of bovine coronavirus (BCoV), bovine rotavirus, and bovine viral diarrhea virus in the feces of normal and diarrheic Korean native calves aged 1-81 days between April and October of 2016 in the Republic of Korea. Methods: Samples were obtained from 50 normal and 93 diarrheic (56 semi-formed, 28 loose, and 9 watery feces) calves in six different regions of northern and southern Korea. These fecal samples were tested for BCoV, bovine rotavirus, and bovine viral diarrhea virus by RT-PCR. Results: Among the three pathogens examined, infection with BCoV was especially prominent in relation to diarrhea among calves aged 1-21 days [odds ratio (OR)=9.3, 95% confidence interval (CI): 1.1-78.9; P=0.02). Infection with BCoV alone (OR=2.9; 95% CI: 1.1-7.6; P=0.03) or co-infection of BCoV with bovine viral diarrhea virus (OR=3.6; 95% CI: 1.0-12.4; P=0.04) was significantly associated with the development of loose feces. Grazing and colostrum intake strongly reduced the occurrence of diarrhea as compared to housed calves (OR=0.2; 95% CI: 0.1-0.4; P=0.00) and calves that had not been fed colostrum (OR=0.2; 95% CI: 0.1-0.7; P=0.02), respectively. Conclusions: The present study suggests that BCoV is involved in calf diarrhea in the Republic of Korea. Therefore, grazing and colostrum intake is recommended for preventing and controlling calf diarrhea caused by BCoV.

14-18

Authors: Weerayuth Saelim, Wichai Santimaleeworagun, Sudaluck Thunyaharn, Dhitiwat Changpradub, Piraporn Juntanawiwat

Objective: To study the minimum inhibitory concentration (MIC) of sulbactam against carbapenem-resistant Acinetobacter baumannii (CR-AB) and to determine the dosage regimens reaching target time of free drug concentration remaining above the MIC (fT>MIC). Methods: Clinical isolates of CR-AB from patients admitted to Phramongkutklao Hospital, Thailand from January 2014 to December 2015 were obtained. The MIC of sulbactam for each CR-AB isolate was determined using the agar dilution method. Each sulbactam regimen was simulated using the Monte Carlo technique to calculate the probability of target attainment (PTA) and the cumulative fraction of response (CFR) in critically ill patients. PTA was defined by how likely a specific drug dose was to reach 40% and 60% fT>MIC. The CFR was the probability of drug dose covering the MIC range of CR-AB. Dosing regimens reaching above 80% of PTA and CFR, were considered as the optimal dosage for documented and empirical therapy, respectively. Results: A total of 118 CR-AB isolates were included in the study. The percentile at the fiftieth and ninetieth MIC of sulbactam were 64 and 192 μg/mL, respectively. For a MIC of sulbactam of 4 μg/mL, all dosage regimens achieved PTA target. However, only a sulbactam dosage of 12 g intravenous daily using 2-4 h infusion or continuous infusion that covered for isolates with a sulbactam MIC of 96 μg/mL, met the PTA or CFR targets. Conclusions: The MIC of sulbactam against CR-AB is quite high. The sulbactam dose of 12 g/day using prolonged infusion was required to achieve the target fT>MIC for CR-AB treatment.

25-30

Authors: Irda Fidrianny, Hendy Suhendy, Muhamad Insanu

Objective: To determine antioxidant activity and phytochemical content from various tubers extracts of sweet potato (Ipomoea batatas) and to explore the correlation of phytochemical content with their antioxidant activities. Methods: Antioxidant activities were tested using DPPH and FRAP assays. Total phenolic was calculated by Folin-Ciocalteu reagent, flavonoid content by Chang’s method and correlation with their antioxidant activities were analyzed by Pearson’s method. Results: PO2 showed highest antioxidant activity, which had the lowest IC50 DPPH (10.54 μg/mL) and the lowest EC50 FRAP (11.14 μg/mL). PO2 showed the highest total phenolic (11.91 g GAE/100 g) and total flavonoid content (17.83 g QE/100 g). There were significantly negative correlation between total phenolic content and flavonoid content in sample PO with their IC50 DPPH and EC50 FRAP. IC50 DPPH of sample PP and PO showed significantly positive correlation with their EC50 FRAP. Conclusions: Result of DPPH method shows that all different ethyl acetate and ethanolic tubers extracts of four varieties of sweet potato are classified as strong and very strong antioxidant. Result of DPPH and FRAP methods indicates that phenolic and flavonoid compounds in sample PO contributes together to antioxidant activities. Antioxidant activities of sample PP and PO by DPPH method are linear to FRAP method.

37-43

Authors: Nurul Jadid, Byan Arasyi Arraniry, Dewi Hidayati, Kristanti Indah Purwani, Wiwi Wikanta, Sylviana Rosyda Hartanti, Rizka Yuanita Rachman

Objective: To investigate the proximate and mineral composition of the Piper retrofractum (P. retrofractum) vahl. Fruit and to evaluate its total alkaloids, phenol and flavonoid. Methods: The proximate composition of P. retrofractum fruit was ananlyzed using standard protocols according to Indonesian Standard and Association of Official Analytical Chemist. Meanwhile, mineral composition of the fruit was analyzed using inductively coupled plasma-mass spectrometry. Phytochemical screening and quantification were performed using standard protocols according to Harborn and spectrophotometric methods. Results: The results showed that P. retrofractum fruit contained carbohydrate (63.4%), crude protein (11.4%), total ash (4.29%), dietary fiber (28.8%) and total fat (2.97%). The fruit also contained calcium, copper, iron, magnesium, phosphor, potassium, sodium and zinc in different concentrations. Additionally, quinone, sterol, glycosides and alkaloid were detected in both n-hexane and ethyl acetate extracts. Moreover, tannin was presented also in ethylacetate and methanol extracts. Meanwhile, methanol extract contained sterol, glycosides, flavones, tannin and alkaloid. The results also revealed that methanol extract of the fruit contained highest phenol compared to other extract. Finally, small quantity of flavonoid (0.060 0%±0.000 2%) was observed. Conclusions: The overall results show that P. retrofractum contains potential nutritional and phytochemicals values, which support their function for pharmaceutical purposes.

52-58

Authors: Fatai Oladunni Balogun, Anofi Omotayo Tom Ashafa

Objective: To investigate the free radical scavenging, antidiabetic, kinetics of enzyme inhibition and cytotoxic potentials of flavonoid-rich extract of Gazania krebsiana leaves using standard methods. Methods: Antioxidant activity of the flavonoids was investigated at scavenging free radicals such as 1,1-diphenyl-2-picryl hydrazyl, nitric oxide, hydroxyl radical, reducing capabilities, 2,2-azinobis (3-ethylbenzothiazoline-6) sulphonic acid as well as metal chelating capability at different concentrations (125-1 000 μg/mL) while the antidiabetic activity was evaluated via the inhibition and kinetics of carbohydrate digestive enzymes including α-amylase, α-glucosidase, sucrase and maltase. Brine shrimp lethality assay was also employed to examine the cytotoxic effects at various concentrations (125-2 000 μg/mL). Results: The study showed the best antioxidant activity in 2,2-azinobis (3-ethylbenzothiazoline-6) sulphonic acid, hydroxyl radical, nitric oxide and metal chelating with IC50 values of (1 089.00±19.29), (719.40±5.35), (633.10±5.31), (1 116.00±22.92) μg/mL, respectively; as compared with butylated hydroxyl anisole (standard) with IC50 of 1 166, 747, 639 and 1 363 μg/mL, respectively. Similarly, strong inhibition of α-glucosidase as well as moderate inhibition of sucrase and maltase enzyme activities which were significantly (P<0.05) better than acarbose were also observed in the study; though acarbose activity against α-amylase was better than the extract. The kinetics or mode of inhibition of α-amylase by the extract revealed an uncompetitive inhibition, competitive for α-glucosidase and non-competitive inhibitions for both sucrase and maltase, while the result of the lethality assay showed a potent cytotoxic effect with LC50 value of 359 μg/mL. Conclusions: The results obtained from this study are suggestive of the antioxidative, antidiabetic and cytotoxic potentials of flavonoid-rich extract of Gazania krebsiana.

67-72

Authors: Saliou Ngom, Raimundo Cabrera Perez, Ma Anta Mbow, Rokhaya Fall, Saliou Niassy, Andreea Cosoveanu, Serigne Mbacké Diop, El Hadji Barka Ndiaye, Moussoukhoye Diop, Georges Lognay

Objective: To evaluate the insecticide, larvicidal and repellent activity of the essential oils from Callistemon viminalis, Melaleuca leucadendron, and Hyptis suaveolens against Chrysodeixis chalcites and to compare it with neem oil (Azadirachta indica). Methods: The essential oils of the leaves of these aromatiques plants were extracted by steam distillation and contacts tests were carried out. Results: Essential oils in ethanol from Callistemon viminalis showed a higher biological activity than the neem with 100% larval mortality at the concentration of 2 μg/mL for 6 h, 100% and 90% in ethanol from Melaleuca leucadendron and Hyptis suaveolens, respectively at the concentration of 4 μg/mL for 24 h. By inhalation, the essential oils from Melaleuca leucadendron and of Hyptis suaveolens were more effective with mortality rates of larvae 100% and 50% respectively at 2 μg/L air applied after 24 h. Nevertheless, the neem has shown to be a repulsive plant and anti-nutritional plant. A significant difference in the percentages of consumption between leaves treated with neem oil and the control samples was observed (Newman-Keuls test) except for Melaleuca leucadendron. Conclusions: The results of the study highlight remarkable biocide, properties of tested extracts, which provides important opportunities for the development of biopesticides.

79-84

Authors: Bhagavathi Sundaram Sivamaruthi, Periyanaina Kesika, Chaiyavat Chaiyasut

Rice bran (RB) is one of the nutrient-rich agricultural byproducts. It is a composite of carbohydrates, lipids, proteins, fibers, minerals, and trace elements such as phosphorus, potassium, magnesium, calcium and manganese. The extraction and purification process influences the quality and quantity of rice bran oil, which is rich in tocopherols, tocotrienols, γ-oryzanol, and unsaturated fatty acids. The bioactive components of RB have been reported for exhibiting antioxidant, anti-inflammatory, hypocholesterolemic, anti-cancer, anti-colitis, and antidiabetic properties. In vitro and in vivo studies, and clinical trials in human volunteers revealed the anti-hyperglycemic activity of RB derived compounds. An updated comprehensive review on the antidiabetic property of RB and its derivative is required to appraise the current knowledge in the particular field. Thus, the present paper covered the composition and bioactivities of RB, and influence of extraction methods on the biological property of rice bran oil and rice bran extract. And the current review also focused on the reported anti-hyperglycemia activity of rice bran derivatives, and its probable mechanism.