Asian Pacific Journal of Tropical Biomedicine

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.

1055–1061

Authors: Natharinee Horata, Kiattawee Choowongkomon, Siriluk Ratanabunyong, Jarinee Tongshoob, Srisin Khusmith

Objectives To explore whether individuals infected with Plasmodium falciparum (P. falciparum) develop antibodies directed against PfEMP1-DBLα, and to assess their IgG subclass distribution in severe and uncomplicated malaria. Methods The anti-PfDBLα IgG and their IgG subclass distributions in plasma of severe (SM) and uncomplicated malaria (UCM) were assessed by enzyme-linked immunoabsorbent assay. The antibody profiles to P. falciparum blood stage antigens were evaluated. CD36 binding ability was determined by static receptor-binding assays. Rosette formation was performed by staining with acridine orange. Results Significantly higher number of UCM (86.48%) than SM (57.78%) plasma contained total acquisition of specific IgG to P. falciparum antigens (P = 0.000). Similar manners were seen in response to P. falciparum DBLα with significant difference (UCM, 59.46% vs SM, 40.00%; P = 0.014). Anti-PfDBLα-IgG1 and -IgG3 were the predominant subclasses. Similar percentage of UCM (31.82%) and SM (33.33%) plasma contained only IgG1, while 13.64% of UCM and 27.78% of SM plasma contained only IgG3. Anti-PfDBLα-IgG1 coexpressed with more than one subclass was noted (UCM, 27.27%; SM, 16.67%). Obviously, IgG1 coexpressed with IgG3 (9.09%) was observed in only UCM plasma. IgG1 was coexpressed with IgG2 in UCM (9.09%) and SM (11.11%) plasma, while IgG1 was coexpressed with IgG4 only in UCM plasma (4.55%). IgG subclasses to P. falciparum antigens were distributed in a similar manner. Only the levels of IgG1, but not IgG3 were significantly higher in UCM than in SM. Conclusions These data suggest that individuals infected with P. falciparum can develop the anti-PfEMP1 antibodies with the major contribution of specific IgG subclasses. The balance and the levels of anti-PfDBLα IgG subclasses play a crucial role in antibody mediated protection against severe malaria.

1067–1070

Authors: Ahmed Tabbabi, Jabeur Daaboub

Objective To evaluate the deltamethrin pyrethroid insecticides against Anopheles labranchiae, potential malaria vector in Tunisia. Methods Six field populations of Anopheles labranchiae mosquitoes were collected from six localities in Northern and Central Tunisia between October and November 2016. Different bioassays were performed to estimate the level of resistance in each collected population. Two synergists were used to estimate the involvement of detoxification enzymes in insecticide resistance. Results All studied strains were resistant and the RR50 ranged from 12.5 in sample #1 to 72.5 in sample #6. Synergist tests using piperonyl butoxide indicated the involvement of monoxygenases enzymes in the recorded resistance. In contrast, the increase of deltamethrin mortality was not significant in presence of S,S,sributyl phosphorothioate (0.8 < SR < 1.2), suggesting no role of esterases (and/or GST) in the resistance phenotype. The correlation recorded between mortality due to DDT and the LC50 of deltamethrin insecticide indicated an insensitive sodium channel affected by Kdr mutation (Spearman rank correlation, r = −0.59, P < 0.01). Conclusions These results should be considered in the current mosquitoes control programs in Tunisia. The use of pesticides and insecticides by both agricultural and public health departments in Tunisia should be more rational to reduce the development of resistance in populations. Different insecticide applications should be implemented alternately.

1079–1084

Authors: Olamide Olajusi Crown, Olanrewaju Sam Olayeriju, Ayodele Oluwaseyi Kolawole, Afolabi Clement Akinmoladun, Mary Tolulope Olaleye, Afolabi Akintunde Akindahunsi

Objective To explore the possible inhibitory potentials and mechanism by Mobola plum (Parinari curatellifolia) seeds crude methanol (CE) and flavonoid-rich (FE) extracts on angiotensin-1-converting enzyme (ACE Ⅰ). Methods The sensitivity and kinetic model of inhibition of CE and FE on ACE Ⅰ using N-[3-(2-furyl)-acryloyl]-Phe-Gly-Gly as enzyme substrate for ACE Ⅰ was evaluated by Michealis Menten approach. The inhibition mechanism was explored from Lineweaver–Burk model and IC50 was determined from Cheng–Prusoff empirical analysis. Results The IC50 of CE and FE were 13.54 and 39.38 μg/mL, respectively. Both extracts exhibited mixed type inhibition with the inhibitory constant (Ki) of CE was between 0.38 and 0.37 μg/mL while that of FE showed a two-fold increase (1.62 μg/mL and 0.28 μg/mL). FE on ACE Ⅰ demonstrated positive cooperativity with a Hill's coefficient of 1.89. Conclusions The study reveals the superior ACE Ⅰ inhibitory potential of CE over FE and suggest that mixed inhibition pattern of the enzyme might be the underlying mechanism of antihypertensive activity.

1092–1096

Authors: Kiattiphumi Saengsorn, Ampa Jimtaisong

Objective To determine hydrophilic–lipophilic balance (HLB) value, stability of formulate emulsion and properties of sacha inchi oil. Methods The physiochemical characteristics of sacha inchi oil were first investigated. Free radical scavenging property was studied by DPPH assay. HLB value of sacha inchi oil was experimentally determined by preparing the emulsion using emulsifiers at different HLB value. Sacha inchi oil emulsion was prepared using the obtained HLB and its stability was conducted by centrifugation, temperature cycling, and accelerated stability test. The efficiency of the prepared emulsion was clinically investigated by 15 volunteers. The primary skin irritation was performed using closed patch test. Subjective sensory assessment was evaluated by using 5-point hedonic scale method. Results Peroxide value of sacha inchi oil was 18.40 meq O2/kg oil and acid value was 1.86 KOH/g oil. The major fatty acids are omega-3 (44%), omega-6 (35%) and omega-9 (9%). The vitamin E content was 226 mg/100 g oil. Moreover, sacha inchi oil (167 ppm) and its emulsion showed 85% and 89% DPPH inhibition, respectively. The experimental HLB value of sacha inchi oil was 8.5. The sacha inchi oil emulsion exhibited good stability after stability test. The emulsion was classified as non-irritant after tested by primary skin irritation method. The skin hydration value significantly increased from 38.59 to 45.21 (P < 0.05) after applying sacha inchi oil emulsion for 1 month and the overall product satisfaction of volunteers after use was with score of 4.2. Conclusions This work provides information on HLB value and emulsion properties of sacha inchi oil which is useful for cosmetic and pharmaceutical application.

1107–1115

Authors: Laavanya M. Kumar, Wan Zuhainis Saad, Rosfarizan Mohamad, Raha Abdul Rahim

Objective To investigate the antibacterial effect of selected lactic acid bacteria (LAB) biofilms on the planktonic and biofilm population of methicillin-resistant Staphylococcus aureus (MRSA) (S547). Methods In this study, biofilm-forming LAB were isolated from tairu and kefir. Isolate Y1 and isolate KF were selected based on their prominent inhibition against test pathogens (using spot-on-agar method and agar-well-diffusion assay) and efficient biofilm production (using tissue culture plate method). They were then identified as Lactobacillus casei (L. casei) Y1 and Lactobacillus plantarum (L. plantarum) KF, respectively using 16S rDNA gene sequencing. The influence of incubation time, temperature and aeration on the biofilm production of L. casei Y1 and L. plantarum KF was also investigated using tissue culture plate method. The inhibitory activity of both the selected LAB biofilms was evaluated against MRSA (Institute for Medical Research code: S547) using L. plantarum ATCC 8014 as the reference strain. Results L. casei Y1 showed the highest reduction of MRSA biofilms, by 3.53 log at 48 h while L. plantarum KF records the highest reduction of 2.64 log at 36 h. In inhibiting planktonic population of MRSA (S547), both L. casei Y1 and L. plantarum KF biofilms recorded their maximum reduction of 4.13 log and 3.41 log at 24 h, respectively. Despite their inhibitory effects being time-dependent, both LAB biofilms exhibited good potential in controlling the biofilm and planktonic population of MRSA (S547). Conclusions The results from this study could highlight the importance of analysing biofilms of LAB to enhance their antibacterial efficacy. Preferably, these protective biofilms of LAB could also be a better alternative to control the formation of biofilms by pathogens such as MRSA.

1120–1124

Authors: Hayette Bouabida, Fouzia Tine-djebbar, Samir Tine, Noureddine Soltani

Objective To evaluate the activity of spiromesifen against the most abundant and investigated mosquito species, Culiseta longiareolata Aitken, 1954 (Diptera, Culicidae). Methods Culiseta longiareolata larvae were collected from untreated areas located at Tébessa (Northeast Algeria). A commercial formulation of spiromesifen (Oberon® 240 SC) was tested at different concentrations ranging between 238 and 1 428 μg/L on newly molted fourth-instar larvae under standard laboratory conditions according to Word Health Organization recommendations. The effects were examined on the mortality, the morphometric measurements, two biomarkers (catalase and malondialdehyde), and the biochemical composition of larvae, respectively. Results The compound exhibited insecticidal activity. Moreover, it disturbed growth and several morphological aberrations were observed. It also affected body volume, biomarkers and contents of carbohydrates, lipids and proteins. A marked effect on lipids and malondialdehyde was noted, confirming its primary mode of action on lipid synthesis. Conclusions Spiromesifen appears less potent than other insecticides tested such as the insect growth disruptors.

1129–1150

Authors: Javed Iqbal, Banzeer Ahsan Abbasi, Tariq Mahmood, Sobia Kanwal, Barkat Ali, Sayed Afzal Shah, Ali Talha Khalil

Cancer is a frightful disease and represents one of the biggest health-care issues for the human race and demands a proactive strategy for cure. Plants are reservoirs for novel chemical entities and provide a promising line for research on cancer. Hitherto, being effective, chemotherapy is accompanied by certain unbearable side effects. Nevertheless, plants and plant derived products is a revolutionizing field as these are Simple, safer, eco-friendly, low-cost, fast, and less toxic as compared with conventional treatment methods. Phytochemicals are selective in their functions and acts specifically on tumor cells without affecting normal cells. Carcinogenesis is complex phenomena that involves many signaling cascades. Phytochemicals are considered suitable candidates for anticancer drug development due to their pleiotropic actions on target events with multiple manners. The research is in progress for developing potential candidates (those can block or slow down the growth of cancer cells without any side effects) from these phytochemicals. Many phytochemicals and their derived analogs have been identified as potential candidates for anticancer therapy. Effort has been made through this comprehensive review to highlight the recent developments and milestones achieved in cancer therapies using phytomolecules with their mechanism of action on nuclear and cellular factors. Furthermore, drugs for cancer treatment and their limitations have also been discussed.

967–978

Authors: Mayura Soonwera, Siriporn Phasomkusolsil

Objective To evaluate adulticidal, larvicidal and oviposition deterrent response of the essential oil from dried Zanthoxylum limonella (Z. limonella) fruit against Aedes aegypti (Ae. aegypti) and Culex quinquefasciatus (Cx. quinquefasciatus). Methods Z. limonella oil was tested by biological assays at 1%, 5% and 10% concentrations in ethanol. Adulticidal efficacy was tested against the 2–3 day old adult females. Larvicidal activity was tested against immature stage of mosquitoes. Oviposition deterrence of the oil was evaluated on gravid females. Results The adult mortality was observed after 24 h with the LC50 of 6.0% for Ae. aegypti, and 5.7% for Cx. quinquefasciatus. Larvicidal bioassay was carried out with the 10% Z. limonella oil against immature stages of Ae. aegypti and Cx. quinquefasciatus, which caused 100% mortality after 12 h and 24 h. In the larvicidal experiment, Z. limonella showed effective result at 1%, 5% and 10% concentrations with the values of LT50  Ae. aegypti = 9.78, 5.61, 0.24 h for larvae and LT50 = 64.08, 21.23 h for pupae; Cx. quinquefasciatus had LT50 = 28.46, 20.25, 1.01 h for larvae and LT50 = 67.52, 27.96, 4.11 h for pupae, respectively. Oviposition deterrence of the oil was evaluated on gravid females. In the study, 10% Z. limonella showed 100% repellency for Ae. aegypti and 99.53% for Cx. quinquefasciatus. Likewise, oviposition activity indexes of these oil concentrations were all negative values ranging from–0.89 to −1.00 for Ae. aegypti and–0.64 to–0.99 for Cx. quinquefasciatus. The oviposition activity indexes values revealed that Z. limonella oil has deterrent effect, and it caused a remarkable negative response resulting in very few eggs. Conclusions This result indicates that Z. limonella oil can be used as an effective adulticide, larvicide and oviposition deterrent against Ae. aegypti and Cx. quinquefasciatus.

987–990

Authors: Maysaa El Sayed Zaki, Nermen Abo El Kheir

Objective To determine the prevalence of astrovirus, norovirus, adenovirus in children below five years old with diarrhea by multiplex reverse transcriptase polymerase chain reaction (RT-PCR) along with rotavirus antigen detection by enzyme linked immunosorbant assay. Methods The study was conducted on children below five years old complaining of acute diarrhea. The study included stool examination by molecular method for detection of norovirus, adenovirus and astrovirus by multiplex RT-PCR. Rotavirus antigen was detected in the stool by enzyme linked immunosorbant assay. Results The study included 100 children below 5 years old with acute diarrhea. Multiplex RT-PCR was positive in 34% of the children. The most frequently detected virus was rotavirus (44%), followed by norovirus (30%), adenovirus (20%) and astrovirus (14%). The clinical symptoms were more significantly associated with viral diarrhea such as fever (P = 0.03), bloody diarrhea (P = 0.025), vomiting (P = 0.000 1) and watery diarrheas (P = 0.05). The frequency of diarrhea with viral pathogen was significantly presented in winter season (39.7%). There were significant frequencies of norovirus and adenovirus in age ranging 1–2 years old (P = 0.04, P = 0.01 respectively). Conclusions The present study spotlights on the prevalence of viral pathogens as an important etiology in diarrhea in children below five years old. Astrovirus, norovirus and adenovirus are common along with rotavirus in this group of patients. Multiplex PCR leads to improve the laboratory diagnosis of these viruses along with antigen detection method. Further longitudinal studies are required to evaluate the epidemiological data associated with these viruses and for proper management of such drastic infection.

998–1004

Authors: Piman Pocasap, Natthida Weerapreeyakul, Waraporn Tanthanuch, Kanjana Thumanu

Objectives To evaluate the concentration differences of sulforaphene and sulforaphane at various ages and in different parts of Raphanus sativus L. var. caudatus with respect to their potential cancer preventive effect on HCT116 colon cancer cells. Methods FTIR–ATR and GC–MS were used to characterize the isothiocyanates in the plant extracts followed by HPLC for quantification. Antiproliferation and apoptosis induction were determined by using MTT assay and flow cytometry, respectively. Results The respective rank of anticancer activity of Raphanus sativus were as follows: vegetative (3 week) < older rosette (4 week) < early-bolting (5 week) < senescence (7 week) < late-bolting (6 week). The low to high concentration of sulforaphene and sulforaphane occurred in the same stage order. Conclusions The reproductive parts (flower, pod, and dry seed) of Raphanus sativus have the greatest isothiocyanate concentration, evidenced by a sulforaphene concentration higher than the sulforaphane. This result should inform the selection of the most appropriate harvesting stage and plant part for use as a potential chemopreventive agent.

1010–1013

Authors: Amira S. Khalil, Nawfal R. Hussein, Maida Y. Shamdeen

Objective To investigate the relationship between hepatitis B virus (HBV) positivity and pregnancy outcomes. Also, the association between HBV-related risk factors and HBV status was studied. Methods A total of 100 HBV positive pregnant women were recruited and the pregnancy outcomes were compared with 301 HBV negative women. Blood samples were collected and tested for HBV by HBsAg ELISA. Data were collected for recruited subjects using interview questionnaire. Results Data analysis showed that 51/100 (51%) of the HBV-positive subjects gave a history of HBV in the family which was significantly higher than that of HBV-negative patients [41/301 (13.6%) P = 0.001]. A significant association was found between positive history of surgery and HBsAg positivity (P = 0.009). Then, pregnancy outcomes were stratified according to the HBV positivity. No significant association was found between HBV status and pregnancy outcomes (P > 0.05 for all). Conclusions Positive family history of HBV and previous surgical procedures are associated with higher rate of HBV positivity. No association is found between HBV positivity and pregnancy outcomes.

1023–1024

Authors: Venu Gopal Jonnalagadda, Kanchan Choudhary, Vijay Kranti Matety

None

1031–1035

Authors: Mahalakshmi Ansari, Gino A. Kurian

Objective To study the renal toxic effect of titanium dioxide nanoparticles (TiNPs) prepared by chemical and green route. Methods TiNPs were prepared by chemical (sol gel technique) and green route (using aqueous extract of Desmodium gangeticum root by using titanium tetraisopropoxide as precursor). Thus prepared TiNPs were characterized using UV–visible spectrophotometry, X-ray diffractometry and evaluated its renal toxic impact in different experimental models viz., Wistar rats (100 mg/kg b.wt.; oral), LLC-PK1 cells (100 mg/mL) and isolated renal mitochondria (0.25, 0.5 and 1 mg/mL). Results Compared to the chemically synthesized TiNPs, Desmodium gangeticum synthesized nanoparticles showed less nephrotoxicity, determined by elevated serum renal markers like urea (62%), creatinine (35%), aspartate aminotransferase (61%) and alanine transaminase (37%) and the result was in agreement with cellular toxicity (measured by MTT assay and lactate dehydrogenase activity). Further toxicity evaluation at the level of mitochondria showed not much significant difference in TiNPs effect between two synthetic routes. Conclusions The biochemical findings in renal tissue and epithelial cell (LLC-PK1) supported by histopathology examination and isolated mitochondrial activity showed minor toxicity with TiNPs prepared by green route (TiNP DG) than TiNP Chem.

1041–1045

Authors: Kaviyarasi Renu, V.G. Abilash, P.B. Tirupathi Pichiah, Thabassum Akthar Syeda, Sankarganesh Arunachalam

Diabetic cardiomyopathy is one of the life threatening complications of diabetes. A number of animal models are being used for studying diabetic cardiomyopathy. In laboratory animal models, induction of cardiomyopathy happens in two stages: first being the induction of diabetic condition and the second being the induction of cardiomyopathy by prolonging diabetic condition. It takes a longer time to develop diabetes with the limited success rate for development of cardiomyopathy. Adriamycin is an effective anti-cancer drug limited by its major side-effect cardiomyopathy. A number of features of Adriamycin treatment mimics diabetes. We postulate that Adriamycin-induced cardiomyopathy might be used as a model system to study diabetic cardiomyopathy in rodents since a number of features of both the cardiomyopathies overlap. Left ventricular hypertrophy, systolic and diastolic dysfunction, myofibrillar loss, and fibrosis are hallmarks of both of the cardiomyopathies. At the molecular level, calcium signaling, endoplasmic reticulum stress, advance glycation endproduct activation, mitochondrial dysfunction, inflammation, lipotoxicity and oxidative stress are similar in both the cardiomyopathies. The signature profile of both the cardiomyopathies shares commonalities. In conclusion, we suggest that Adriamycin induced cardiomyopathic animal model can be used for studying diabetic cardiomyopathy and would save time for researchers working on cardiomyopathy developed in rodent using the traditional method.