Search

Research has confirmed that the utilisation of Antarctic microorganisms, such as bacteria, yeasts and fungi, in the bioremediation of diesel may provide practical alternative approaches. However, to date there has been very little attention towards Antarctic microalgae as potential hydrocarbon degraders. Therefore, this study focused on the utilisation of an Antarctic microalga in the bioremediation of diesel. The studied microalgal strain was originally obtained from a freshwater ecosystem in Paradise Bay, western Antarctic Peninsula. When analysed in systems with and without aeration, this microalgal strain achieved a higher growth rate under aeration. To maintain the growth of this microalga optimally, a conventional one-factor-at a-time (OFAT) analysis was also conducted. Based on the optimized parameters, algal growth and diesel degradation performance was highest at pH 7.5 with 0.5 mg/L NaCl concentration and 0.5 g/L of NaNO3 as a nitrogen source. This currently unidentified microalga flourished in the presence of diesel, with maximum algal cell numbers on day 7 of incubation in the presence of 1% v/v diesel. Chlorophyll a, b and carotenoid contents of the culture were greatest on day 9 of incubation. The diesel degradation achieved was 64.5% of the original concentration after 9 days. Gas chromatography analysis showed the complete mineralisation of C7–C13 hydrocarbon chains. Fourier transform infrared spectroscopy analysis confirmed that strain WCY_AQ5_3 fully degraded the hydrocarbon with bioabsorption of the products. Morphological and molecular analyses suggested that this spherical, single-celled green microalga was a member of the genus Micractinium. The data obtained confirm that this microalga is a suitable candidate for further research into the degradation of diesel in Antarctica.

The bone morphogenic protein (BMP) family is a member of the TGF-beta superfamily and plays a crucial role during the onset of gut inflammation and arthritis diseases. Recent studies have reported a connection with the gut–joint axis; however, the genetic players are still less explored. Meanwhile, BDMC33 is a newly synthesized anti-inflammatory drug candidate. Therefore, in our present study, we analysed the genome-wide features of the BMP family as well as the role of BMP members in gut-associated arthritis in an inflammatory state and the ability of BDMC33 to attenuate this inflammation. Firstly, genome-wide analyses were performed on the BMP family in the zebrafish genome, employing several in silico techniques. Afterwards, the effects of curcumin analogues on BMP gene expression in zebrafish larvae induced with TNBS (0.78 mg/mL) were determined using real time-qPCR. A total of 38 identified BMP proteins were revealed to be clustered in five major clades and contain TGF beta and TGF beta pro peptide domains. Furthermore, BDMC33 suppressed the expression of four selected BMP genes in the TNBS-induced larvae, where the highest gene suppression was in the BMP2a gene (an eight-fold decrement), followed by BMP7b (four-fold decrement), BMP4 (four-fold decrement), and BMP6 (three-fold decrement). Therefore, this study reveals the role of BMPs in gut-associated arthritis and proves the ability of BDMC33 to act as a potential anti-inflammatory drug for suppressing TNBS-induced BMP genes in zebrafish larvae.

The bone morphogenic protein (BMP) family is a member of the TGF-beta superfamily and plays a crucial role during the onset of gut inflammation and arthritis diseases. Recent studies have reported a connection with the gut–joint axis; however, the genetic players are still less explored. Meanwhile, BDMC33 is a newly synthesized anti-inflammatory drug candidate. Therefore, in our present study, we analysed the genome-wide features of the BMP family as well as the role of BMP members in gut-associated arthritis in an inflammatory state and the ability of BDMC33 to attenuate this inflammation. Firstly, genome-wide analyses were performed on the BMP family in the zebrafish genome, employing several in silico techniques. Afterwards, the effects of curcumin analogues on BMP gene expression in zebrafish larvae induced with TNBS (0.78 mg/mL) were determined using real time-qPCR. A total of 38 identified BMP proteins were revealed to be clustered in five major clades and contain TGF beta and TGF beta pro peptide domains. Furthermore, BDMC33 suppressed the expression of four selected BMP genes in the TNBS-induced larvae, where the highest gene suppression was in the BMP2a gene (an eight-fold decrement), followed by BMP7b (four-fold decrement), BMP4 (four-fold decrement), and BMP6 (three-fold decrement). Therefore, this study reveals the role of BMPs in gut-associated arthritis and proves the ability of BDMC33 to act as a potential anti-inflammatory drug for suppressing TNBS-induced BMP genes in zebrafish larvae.

Diseases caused by oxidative stress can be prevented by antioxidant. Current treatments for those neurodegenerative diseases are not effective and cause many side effects. Thus, the search for alternative medicines is in high demand. Therefore, the main purposed of this study is to evaluate the neuroprotective effects of Curcuma longa (rhizome) 80% methanol extract. Antioxidant using dichlorofuoresence diacetate (DCF-DA) assay on SH-SY5Y cells revealed high activities of Curcuma longa (rhizome) extract with IC50 of 105.9±0.8 µg/mL. Sub-acute and chronic toxicity tests of the plant extract on adult Javanese medaka (Oryzias javanicus) showed high toxicity effect with LC50 of 24.15±0.8 mg/L and 13.69±0.7 mg/L respectively. Neuroprotective tests using cholinesterase assay disclose significant differences at P<0.05 between the group that are exposed to arsenic and treated with the crude extract and the group that are exposed to only arsenic. Identification of vitexin and isovitexin justified the high antioxidant potential of this plant leaf and it highest benefit to be used as medicinal supplement.

Honey is prone to be adulterated through mixing with sugars, cheap and low-quality honey, and other adulterants. Consumption of adulterated honey may cause several health issues such as weight gain, diabetes, and liver and kidney dysfunction. Therefore, studying the impact of consumption of adulterated honey on consumers is critical since there is a lack of study in this field. Hence, the aims of this paper were: (1) to determine the lethal concentration (LC50) of adulterated honey using zebrafish embryo, (2) to elucidate toxicology of selected adulterated honey based on lethal dose (LD50) using adult zebrafish, (3) to determine the effects of adulterated honey on histological changes of zebrafish, and (4) to screen the metabolites profile of adulterated honey by using zebrafish blood serum. The LC50 of Heterotrigona itama honey (acacia honey) and its sugar adulterants (light corn sugar, cane sugar, inverted sugar, and palm sugar in the proportion of 1–3% (w/w) from the total volume) was determined by the toxicological assessment of honey samples on zebrafish embryos (different exposure concentrations in 24, 48, 72, and 96 h postfertilization (hpf)). Pure H. itama honey represents the LC50 of 34.40 ± 1.84 (mg/mL) at 96 hpf, while the inverted sugar represents the lowest LC50 (5.03 ± 0.92 mg/mL) among sugar adulterants. The highest concentration (3%) of sugar adulterants were used to study the toxicology of adulterated honey using adult zebrafish in terms of acute, prolong-acute, and sub-acute tests. The results of the LD50 from the sub-acute toxicity test of pure H. itama honey was 2.33 ± 0.24 (mg/mL). The histological studies of internal organs showed a lesion in the liver, kidney, and spleen of adulterated treated-honey groups compared to the control group. Furthermore, the LC-MS/MS results revealed three endogenous metabolites in both the pure and adulterated honey treated groups, as follows: (1) S-Cysteinosuccinic acid, (2) 2,3-Diphosphoglyceric acid, and (3) Cy

Diseases caused by oxidative stress can be prevented by antioxidant. Current treatments for those neurodegenerative diseases are not effective and cause many side effects. Thus, the search for alternative medicines is in high demand. Therefore, the main purposed of this study is to evaluate the neuroprotective effects of Curcuma longa (rhizome) 80% methanol extract. Antioxidant using dichlorofuoresence diacetate (DCF-DA) assay on SH-SY5Y cells revealed high activities of Curcuma longa (rhizome) extract with IC50 of 105.9±0.8 µg/mL. Sub-acute and chronic toxicity tests of the plant extract on adult Javanese medaka (Oryzias javanicus) showed high toxicity effect with LC50 of 24.15±0.8 mg/L and 13.69±0.7 mg/L respectively. Neuroprotective tests using cholinesterase assay disclose significant differences at P<0.05 between the group that are exposed to arsenic and treated with the crude extract and the group that are exposed to only arsenic. Identification of vitexin and isovitexin justified the high antioxidant potential of this plant leaf and it highest benefit to be used as medicinal supplement.

Honey is prone to be adulterated through mixing with sugars, cheap and low-quality honey, and other adulterants. Consumption of adulterated honey may cause several health issues such as weight gain, diabetes, and liver and kidney dysfunction. Therefore, studying the impact of consumption of adulterated honey on consumers is critical since there is a lack of study in this field. Hence, the aims of this paper were: (1) to determine the lethal concentration (LC50) of adulterated honey using zebrafish embryo, (2) to elucidate toxicology of selected adulterated honey based on lethal dose (LD50) using adult zebrafish, (3) to determine the effects of adulterated honey on histological changes of zebrafish, and (4) to screen the metabolites profile of adulterated honey by using zebrafish blood serum. The LC50 of Heterotrigona itama honey (acacia honey) and its sugar adulterants (light corn sugar, cane sugar, inverted sugar, and palm sugar in the proportion of 1–3% (w/w) from the total volume) was determined by the toxicological assessment of honey samples on zebrafish embryos (different exposure concentrations in 24, 48, 72, and 96 h postfertilization (hpf)). Pure H. itama honey represents the LC50 of 34.40 ± 1.84 (mg/mL) at 96 hpf, while the inverted sugar represents the lowest LC50 (5.03 ± 0.92 mg/mL) among sugar adulterants. The highest concentration (3%) of sugar adulterants were used to study the toxicology of adulterated honey using adult zebrafish in terms of acute, prolong-acute, and sub-acute tests. The results of the LD50 from the sub-acute toxicity test of pure H. itama honey was 2.33 ± 0.24 (mg/mL). The histological studies of internal organs showed a lesion in the liver, kidney, and spleen of adulterated treated-honey groups compared to the control group. Furthermore, the LC-MS/MS results revealed three endogenous metabolites in both the pure and adulterated honey treated groups, as follows: (1) S-Cysteinosuccinic acid, (2) 2,3-Diphosphoglyceric acid, and (3) Cy

Background: Aripiprazole, which is a quinolinone derivative, has been widely used to treat schizophrenia, major depressive disorder, and bipolar disorder. Purpose: A Central Composite Rotatable Design (CCRD) of Response Surface Methodology (RSM) was used purposely to optimize process parameters conditions for formulating nanoemulsion containing aripiprazole using high emulsification methods. Methods: This design is used to investigate the influences of four independent variables (overhead stirring time (A), shear rate (B), shear time (C), and the cycle of high-pressure homogenizer (D)) on the response variable namely, a droplet size (Y) of nanoemulsion containing aripiprazole. Results: The optimum conditions suggested by the predicted model were: 120 min of overhead stirring time, 15 min of high shear homogenizer time, 4400 rpm of high shear homogenizer rate and 11 cycles of high-pressure homogenizer, giving a desirable droplet size of nanoemulsion containing aripiprazole of 64.52 nm for experimental value and 62.59 nm for predicted value. The analysis of variance (ANOVA) showed the quadratic polynomial fitted the experimental values with F-value (9.53), a low p-value (0.0003) and a non-significant lack of-fit. It proved that the models were adequate to predict the relevance response. The optimized formulation with a viscosity value of 3.72 mPa.s and pH value of 7.4 showed good osmolality value (297 mOsm/kg) and remained stable for three months in three different temperatures (4°C, 25°C, and 45°C). Conclusion: This proven that response surface methodology is an efficient tool to produce desirable droplet size of nanoemulsion containing aripiprazole for parenteral delivery application.

Background: Aripiprazole, which is a quinolinone derivative, has been widely used to treat schizophrenia, major depressive disorder, and bipolar disorder. Purpose: A Central Composite Rotatable Design (CCRD) of Response Surface Methodology (RSM) was used purposely to optimize process parameters conditions for formulating nanoemulsion containing aripiprazole using high emulsification methods. Methods: This design is used to investigate the influences of four independent variables (overhead stirring time (A), shear rate (B), shear time (C), and the cycle of high-pressure homogenizer (D)) on the response variable namely, a droplet size (Y) of nanoemulsion containing aripiprazole. Results: The optimum conditions suggested by the predicted model were: 120 min of overhead stirring time, 15 min of high shear homogenizer time, 4400 rpm of high shear homogenizer rate and 11 cycles of high-pressure homogenizer, giving a desirable droplet size of nanoemulsion containing aripiprazole of 64.52 nm for experimental value and 62.59 nm for predicted value. The analysis of variance (ANOVA) showed the quadratic polynomial fitted the experimental values with F-value (9.53), a low p-value (0.0003) and a non-significant lack of-fit. It proved that the models were adequate to predict the relevance response. The optimized formulation with a viscosity value of 3.72 mPa.s and pH value of 7.4 showed good osmolality value (297 mOsm/kg) and remained stable for three months in three different temperatures (4°C, 25°C, and 45°C). Conclusion: This proven that response surface methodology is an efficient tool to produce desirable droplet size of nanoemulsion containing aripiprazole for parenteral delivery application.

Wan Sarah Samiun,1 Siti Efliza Ashari,1,2 Norazlinaliza Salim,1,2 Syahida Ahmad3 1Integrated Chemical Biophysics Research, Faculty of Science, Universiti Putra Malaysia, Serdang 43400, Selangor, Malaysia; 2Centre of Foundation Studies for Agricultural Sciences, Universiti Putra Malaysia, Serdang 43400, Selangor, Malaysia; 3Department of Biochemistry, Faculty of Biotechnology and Biomolecular Sciences, Universiti Putra Malaysia, Serdang 43400, Selangor, MalaysiaCorrespondence: Wan Sarah Samiun; Siti Efliza AshariIntegrated Chemical Biophysics Research, Faculty of Science, Universiti Putra Malaysia, Serdang 43400, Selangor, MalaysiaTel +603-8946 6997Email ctefliza@upm.edu.my; wansarahsamiun@yahoo.comBackground: Aripiprazole, which is a quinolinone derivative, has been widely used to treat schizophrenia, major depressive disorder, and bipolar disorder.Purpose: A Central Composite Rotatable Design (CCRD) of Response Surface Methodology (RSM) was used purposely to optimize process parameters conditions for formulating nanoemulsion containing aripiprazole using high emulsification methods.Methods: This design is used to investigate the influences of four independent variables (overhead stirring time (A), shear rate (B), shear time (C), and the cycle of high-pressure homogenizer (D)) on the response variable namely, a droplet size (Y) of nanoemulsion containing aripiprazole.Results: The optimum conditions suggested by the predicted model were: 120 min of overhead stirring time, 15 min of high shear homogenizer time, 4400 rpm of high shear homogenizer rate and 11 cycles of high-pressure homogenizer, giving a desirable droplet size of nanoemulsion containing aripiprazole of 64.52 nm for experimental value and 62.59 nm for predicted value. The analysis of variance (ANOVA) showed the quadratic polynomial fitted the experimental values with F-value (9.53), a low p-value (0.0003) and a non-significant lack of-fit. It proved that the models were adequate to predict the relevance

Sabah snake grass or Clinacanthus nutans has drawn public interest having significant economic benefits attributable to the presence of phytochemicals and several interesting bioactive constituents that may differ according to harvesting age and harvesting frequency. The current study was aimed to evaluate the effect of harvesting age and harvesting frequency towards herbal yield, antioxidant activities, phytochemicals synthesis, and bioactive compounds of C. nutans. A factorial randomized completely block design with five replications was used to illustrate the relationship between herbal yield, DPPH (2, 2-diphenyl-1-picrylhydrazyl) and ferric reducing antioxidant power (FRAP) assays, total phenolic and flavonoid content affected by harvesting age (week 8, 12, and 16 after transplanting), and harvesting frequency (harvest 1, 2, and 3). The bioactive compounds by HPLC were also determined to describe the interaction effect between both harvesting age and harvesting frequency. The yield, antioxidant activities, and phytochemical contents were gradually increased as the plant grew, with the highest recorded during week 16. However, the synthesis and activities of phytochemicals were reduced in subsequent harvests despite the increment of the herbal yield. All bioactive compounds were found to be influenced insignificantly and significantly by harvesting age and harvesting frequency, respectively, specifically to shaftoside, iso-orientin, and orientin. Among all constituents, shaftoside was the main compound at various harvesting ages and harvesting frequencies. These results indicated that harvesting at week 16 with 1st harvest frequency might enhance the yield while sustaining the high synthesis of polyphenols and antioxidant activities of C. nutans.

Sabah snake grass or Clinacanthus nutans has drawn public interest having significant economic benefits attributable to the presence of phytochemicals and several interesting bioactive constituents that may differ according to harvesting age and harvesting frequency. The current study was aimed to evaluate the effect of harvesting age and harvesting frequency towards herbal yield, antioxidant activities, phytochemicals synthesis, and bioactive compounds of C. nutans. A factorial randomized completely block design with five replications was used to illustrate the relationship between herbal yield, DPPH (2, 2-diphenyl-1-picrylhydrazyl) and ferric reducing antioxidant power (FRAP) assays, total phenolic and flavonoid content affected by harvesting age (week 8, 12, and 16 after transplanting), and harvesting frequency (harvest 1, 2, and 3). The bioactive compounds by HPLC were also determined to describe the interaction effect between both harvesting age and harvesting frequency. The yield, antioxidant activities, and phytochemical contents were gradually increased as the plant grew, with the highest recorded during week 16. However, the synthesis and activities of phytochemicals were reduced in subsequent harvests despite the increment of the herbal yield. All bioactive compounds were found to be influenced insignificantly and significantly by harvesting age and harvesting frequency, respectively, specifically to shaftoside, iso-orientin, and orientin. Among all constituents, shaftoside was the main compound at various harvesting ages and harvesting frequencies. These results indicated that harvesting at week 16 with 1st harvest frequency might enhance the yield while sustaining the high synthesis of polyphenols and antioxidant activities of C. nutans.

Microglial cells are the primary immune cell resident in the brain. Growing evidence indicates that microglial cells play a prominent role in alcohol-induced brain pathologies. However, alcohol-induced effects on microglial cells and the underlying mechanisms are not fully understood, and evidence exists to support generation of oxidative stress due to NADPH oxidases (NOX_-mediated production of reactive oxygen species (ROS). Here, we investigated the role of the oxidative stress-sensitive Ca2+-permeable transient receptor potential melastatin-related 2 (TRPM2) channel in ethanol (EtOH)-induced microglial cell death using BV2 microglial cells. Like H2O2, exposure to EtOH induced concentration-dependent cell death, assessed using a propidium iodide assay. H2O2/EtOH-induced cell death was inhibited by treatment with TRPM2 channel inhibitors and also treatment with poly(ADP-ribose) polymerase (PARP) inhibitors, demonstrating the critical role of PARP and the TRPM2 channel in EtOH-induced cell death. Exposure to EtOH, as expected, led to an increase in ROS production, shown using imaging of 2’,7’-dichlorofluorescein fluorescence. Consistently, EtOH-induced microglial cell death was suppressed by inhibition of NADPH oxidase (NOX) as well as inhibition of protein kinase C. Taken together, our results suggest that exposure to high doses of ethanol can induce microglial cell death via the NOX/ROS/PARP/TRPM2 signaling pathway, providing novel and potentially important insights into alcohol-induced brain pathologies.

Microglial cells are the primary immune cell resident in the brain. Growing evidence indicates that microglial cells play a prominent role in alcohol-induced brain pathologies. However, alcohol-induced effects on microglial cells and the underlying mechanisms are not fully understood, and evidence exists to support generation of oxidative stress due to NADPH oxidases (NOX_-mediated production of reactive oxygen species (ROS). Here, we investigated the role of the oxidative stress-sensitive Ca2+-permeable transient receptor potential melastatin-related 2 (TRPM2) channel in ethanol (EtOH)-induced microglial cell death using BV2 microglial cells. Like H2O2, exposure to EtOH induced concentration-dependent cell death, assessed using a propidium iodide assay. H2O2/EtOH-induced cell death was inhibited by treatment with TRPM2 channel inhibitors and also treatment with poly(ADP-ribose) polymerase (PARP) inhibitors, demonstrating the critical role of PARP and the TRPM2 channel in EtOH-induced cell death. Exposure to EtOH, as expected, led to an increase in ROS production, shown using imaging of 2’,7’-dichlorofluorescein fluorescence. Consistently, EtOH-induced microglial cell death was suppressed by inhibition of NADPH oxidase (NOX) as well as inhibition of protein kinase C. Taken together, our results suggest that exposure to high doses of ethanol can induce microglial cell death via the NOX/ROS/PARP/TRPM2 signaling pathway, providing novel and potentially important insights into alcohol-induced brain pathologies.

Salmonella enterica serovar (ser.) Enteritidis (S. Enteritidis) is a foodborne pathogen often associated with contaminated poultry products. This study evaluated the anti-adherence and intracellular clearance capability of oligosaccharides extracted from palm kernel cake (PKC), a by-product of the palm oil industry, and compared its efficacy with commercial prebiotics— fructooligosaccharide (FOS) and mannanoligosaccharide (MOS)—against S. Enteritidis in vitro. Based on the degree of polymerization (DP), PKC oligosaccharides were further divided into ‘Small’ (DP ≤ 6) and ‘Big’ (DP > 6) fractions. Results showed that the Small and Big PKC fractions were able to reduce (p < 0.05) S. Enteritidis adherence to Cancer coli-2 (Caco-2) cells at 0.1 mg/ mL while MOS and FOS showed significant reduction at 1.0 mg/mL and 10.0 mg/mL, respectively. In terms of S. Enteritidis clearance, oligosaccharide-treated macrophages showed better S. Enteritidis clearance over time at 50 µg/mL for Small, Big and MOS, while FOS required a concentration of 500 µg/mL for a similar effect. This data highlights that oligosaccharides from PKC, particularly those of lower DP, were more effective than MOS and FOS at reducing S. Enteritidis adherence and enhancing S. Enteritidis clearance in a cell culture model.

An ideal model organism for neurotoxicology research should meet several characteristics, such as low cost and amenable for high throughput testing. Javanese medaka (JM) has been widely used in the ecotoxicological studies related to the marine and freshwater environment, but rarely utilized for biomedical research. Therefore, in this study, the applicability of using JM in the neurotoxicology research was assessed using biochemical comparison with an established model organism, the zebrafish. Identification of biochemical changes due to the neurotoxic effects of ethanol and endosulfan was assessed using Fourier Transform Infrared (FTIR) analysis. Treatment with ethanol affected the level of lipids, proteins, glycogens and nucleic acids in the brain of JM. Meanwhile, treatment with endosulfan showed alteration in the level of lipids and nucleic acids. For the zebrafish, exposure to ethanol affected the level of protein, fatty acid and amino acid, and exposure to endosulfan induced alteration in the fatty acids, amino acids, nucleic acids and protein in the brain of zebrafish. The sensitive response of the JM toward chemicals exposure proved that it was a valuable model for neurotoxicology research. More studies need to be conducted to further develop JM as an ideal model organism for neurotoxicology research.

Salmonella enterica serovar (ser.) Enteritidis (S. Enteritidis) is a foodborne pathogen often associated with contaminated poultry products. This study evaluated the anti-adherence and intracellular clearance capability of oligosaccharides extracted from palm kernel cake (PKC), a by-product of the palm oil industry, and compared its efficacy with commercial prebiotics— fructooligosaccharide (FOS) and mannanoligosaccharide (MOS)—against S. Enteritidis in vitro. Based on the degree of polymerization (DP), PKC oligosaccharides were further divided into ‘Small’ (DP ≤ 6) and ‘Big’ (DP > 6) fractions. Results showed that the Small and Big PKC fractions were able to reduce (p < 0.05) S. Enteritidis adherence to Cancer coli-2 (Caco-2) cells at 0.1 mg/ mL while MOS and FOS showed significant reduction at 1.0 mg/mL and 10.0 mg/mL, respectively. In terms of S. Enteritidis clearance, oligosaccharide-treated macrophages showed better S. Enteritidis clearance over time at 50 µg/mL for Small, Big and MOS, while FOS required a concentration of 500 µg/mL for a similar effect. This data highlights that oligosaccharides from PKC, particularly those of lower DP, were more effective than MOS and FOS at reducing S. Enteritidis adherence and enhancing S. Enteritidis clearance in a cell culture model.

An ideal model organism for neurotoxicology research should meet several characteristics, such as low cost and amenable for high throughput testing. Javanese medaka (JM) has been widely used in the ecotoxicological studies related to the marine and freshwater environment, but rarely utilized for biomedical research. Therefore, in this study, the applicability of using JM in the neurotoxicology research was assessed using biochemical comparison with an established model organism, the zebrafish. Identification of biochemical changes due to the neurotoxic effects of ethanol and endosulfan was assessed using Fourier Transform Infrared (FTIR) analysis. Treatment with ethanol affected the level of lipids, proteins, glycogens and nucleic acids in the brain of JM. Meanwhile, treatment with endosulfan showed alteration in the level of lipids and nucleic acids. For the zebrafish, exposure to ethanol affected the level of protein, fatty acid and amino acid, and exposure to endosulfan induced alteration in the fatty acids, amino acids, nucleic acids and protein in the brain of zebrafish. The sensitive response of the JM toward chemicals exposure proved that it was a valuable model for neurotoxicology research. More studies need to be conducted to further develop JM as an ideal model organism for neurotoxicology research.

Wan Sarah Samiun,1 Siti Efliza Ashari,1,2 Norazlinaliza Salim,1,2 Syahida Ahmad3 1Integrated Chemical Biophysics Research, Faculty of Science, Universiti Putra Malaysia, Serdang 43400, Selangor, Malaysia; 2Centre of Foundation Studies for Agricultural Sciences, Universiti Putra Malaysia, Serdang 43400, Selangor, Malaysia; 3Department of Biochemistry, Faculty of Biotechnology and Biomolecular Sciences, Universiti Putra Malaysia, Serdang 43400, Selangor, MalaysiaCorrespondence: Wan Sarah Samiun; Siti Efliza AshariIntegrated Chemical Biophysics Research, Faculty of Science, Universiti Putra Malaysia, Serdang 43400, Selangor, MalaysiaTel +603-8946 6997Email ctefliza@upm.edu.my; wansarahsamiun@yahoo.comBackground: Aripiprazole, which is a quinolinone derivative, has been widely used to treat schizophrenia, major depressive disorder, and bipolar disorder.Purpose: A Central Composite Rotatable Design (CCRD) of Response Surface Methodology (RSM) was used purposely to optimize process parameters conditions for formulating nanoemulsion containing aripiprazole using high emulsification methods.Methods: This design is used to investigate the influences of four independent variables (overhead stirring time (A), shear rate (B), shear time (C), and the cycle of high-pressure homogenizer (D)) on the response variable namely, a droplet size (Y) of nanoemulsion containing aripiprazole.Results: The optimum conditions suggested by the predicted model were: 120 min of overhead stirring time, 15 min of high shear homogenizer time, 4400 rpm of high shear homogenizer rate and 11 cycles of high-pressure homogenizer, giving a desirable droplet size of nanoemulsion containing aripiprazole of 64.52 nm for experimental value and 62.59 nm for predicted value. The analysis of variance (ANOVA) showed the quadratic polynomial fitted the experimental values with F-value (9.53), a low p-value (0.0003) and a non-significant lack of-fit. It proved that the models were adequate to predict the relevance

A study was conducted to establish in vitro culture conditions for maximum production of biomass and flavonoid content for Ficus deltoidea var. kunstleri, locally named as Mas Cotek, known to have a wide variety of potential beneficial attributes for human health. Size of initial inoculum, cell aggregate and initial pH value have been suggested to influent content of biomass and flavonoid for cell suspension culture in several plant species. In the present study, leaf explants were cultured by cell suspension culture procedures in MSB5 basal medium supplemented with predetermined supplements of 30 g/L sucrose, 2.75 g/L gelrite, 2 mg/L picloram and 1 mg/L kinetin with continuous agitation of 120 rpm in a standard laboratory environment. Establishment of cell suspension culture was accomplished by culturing resulting callus in different initial fresh weight of cells (0.10, 0.25, 0.50, 1.0, and 2.0 g/25 mL of media) using similar basal medium. The results showed that the highest production of biomass (0.65 g/25 mL of media) was recorded from an initial inoculum size of 2.0 g/25 mL media, whereas the highest flavonoid (3.3 mg RE/g DW) was found in 0.5 g/25 mL of media. Cell suspension fractions classified according to their sizes (500–750 µm, 250–500 µm, and <250 µm). Large cell aggregate size (500–750 µm) cultured at pH 5.75 produced the highest cell biomass (0.28 g/25 mL media) and flavonoid content (3.3 mg RE/g DW). The study had established the optimum conditions for the production of total antioxidant and flavonoid content using DPPH and FRAP assays in cell suspension culture of F. deltoidea var. kunstleri.

A study was conducted to establish in vitro culture conditions for maximum production of biomass and flavonoid content for Ficus deltoidea var. kunstleri, locally named as Mas Cotek, known to have a wide variety of potential beneficial attributes for human health. Size of initial inoculum, cell aggregate and initial pH value have been suggested to influent content of biomass and flavonoid for cell suspension culture in several plant species. In the present study, leaf explants were cultured by cell suspension culture procedures in MSB5 basal medium supplemented with predetermined supplements of 30 g/L sucrose, 2.75 g/L gelrite, 2 mg/L picloram and 1 mg/L kinetin with continuous agitation of 120 rpm in a standard laboratory environment. Establishment of cell suspension culture was accomplished by culturing resulting callus in different initial fresh weight of cells (0.10, 0.25, 0.50, 1.0, and 2.0 g/25 mL of media) using similar basal medium. The results showed that the highest production of biomass (0.65 g/25 mL of media) was recorded from an initial inoculum size of 2.0 g/25 mL media, whereas the highest flavonoid (3.3 mg RE/g DW) was found in 0.5 g/25 mL of media. Cell suspension fractions classified according to their sizes (500–750 µm, 250–500 µm, and <250 µm). Large cell aggregate size (500–750 µm) cultured at pH 5.75 produced the highest cell biomass (0.28 g/25 mL media) and flavonoid content (3.3 mg RE/g DW). The study had established the optimum conditions for the production of total antioxidant and flavonoid content using DPPH and FRAP assays in cell suspension culture of F. deltoidea var. kunstleri.

Nigella sativa L. seed, also recognized as black cumin is a medicinal plant that possesses miraculous power of healing due to the abilities to cure various ailments. Owing to the oily properties of plant oil, the oil ester is more preferable exhibiting special features such as non-toxic, outstanding moisturizing action and most importantly the absence of oily texture. In this study, Nigella sativa oil-based ferulate ester was synthesized via transesterification of Nigella sativa oil (NSO) and ethyl ferulate (EF) in the presence of immobilized lipase, Lipozyme RM IM from Rhizomucor miehei. A five-level-four-factor central composite rotatable design (CCRD) from response surface methodology (RSM) was applied to study the influence of synthesis parameters: lipase dosage (50-80 mg), temperature (40-60°C), substrate ratio of NSO:EF (2.5:1-5.5:1 w/w) and time (3-7 hr) aiming for the maximum percentage yield of ester. The optimized synthesis conditions were lipase dosage of 67 mg, temperature of 56°C, substrate ratio (NSO:EF) of 4.4:1 w/w and time of 4 hr. The maximum production of ester obtained was 49.87% which is acceptable with the predicted value of 48.82%.

Legumes have become increasingly in demand due to the rich nutrient compositions and phytochemicals of pulses. However, the seed coats of some legume food products were removed prior consumption causing the food loss its nutritional value. The red kidney bean, RKB (Phaseolus vulgaris) and chickpea, CP (Cicer arietenum) which are the common beans among population were investigated in this study. Seed coats of these beans were analysed for the nutritional composition, phenolic compound and antioxidant properties. Carbohydrate was the major macronutrient in both seed coats. RKB and CP seed coats showed statistically significant composition of moisture, fat, protein and fibre. The caloric value of RKB seed coat (2.63 kcal/g) is higher than CP seed coat (2.29 kcal/g). Nevertheless, CP seed coat is a better source of fibre (27%) than RKB seed coat. Total phenolic content (TPC) of RKB seed coat was 12.14 mg GAE/g, which is much higher than in CP seed coat (0.25 mg GAE/g). Interestingly, the seed coat of RKB has strong antioxidant potency with DPPH assay (IC50 = 105.18 µg/ml) comparable to standard Trolox (IC50 = 96.42 µg/ml), which is much lower than the seed coat of CP (IC50 = 606.12 µg/ml). In addition, the antioxidant activity was highly correlated with TPC content of both seed coats. These properties make the seed coat of both beans are excellent candidates of potent nutraceutical.

In vitro cell viability tests are usually done using 3-(4,5 dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTS) method. A new spectrofluorometry method was developed using acridine orange (AO) and propidium iodide (PI) using multi-label microplate reader. Nine biogenic amines (BAs) [histamine (HIM), putrescine (PUT), cadaverine (CAD), 2-phenylethylamine (PHM), tyramine (TYM), tryptamine (TPM), spermine (SPM), spermidine (SPD) and agmatine (AGM)] were exposed to RAW 264.7 macrophage in singles at 37°C with 5% carbon dioxide supplementation for 18–24 hours and cell viability was determined using MTS method and AO/PI developed method using dual-spectrofluorometry. Based on MTS assay, SPM and SPD were found to be cytotoxic and it was supported by AO/PI assay. The precedence of disintegration in the nucleus rather than mitochondria upon cell non-viability was also supported by transmission electron microscopy (TEM). The results showed that AO/PI method could be used as an alternative method to determine cytotoxicity besides usual usage in confocal microscopy.

Jatropha curcas is a medicinal plant with many therapeutic properties such as anti-inflammatory, anti-malaria, anti-cancer and antioxidant. The root extract has been shown to possess high anti-inflammatory activity. Previously, the compounds responsible for this activity have not been fully elucidated. Two fractions (Fraction 1 and Fraction 2) obtained from a preparative HPLC of the root extract showed significant anti-inflammatory and cytotoxic activities in RAW 264.7 murine macrophage cells with Fraction 1 giving higher nitric oxide (NO) inhibition compared to Fraction 2 and L-NAME. Further purification steps involving column chromatography, thin layer chromatography and analytical HPLC of Fraction 1 produced two fractions labeled as Fraction A and Fraction B. Both fractions showed anti-inflammatory activity without cytotoxic activity in RAW 264.7 cells. Liquid chromatography tandem mass spectrometry (LC-MS/MS) analysis showed that Fraction A contained a group of 18 carbon fatty acid compounds consisting of 2 oxooctadecanoic acids; 15, 16 dihydroxy 9Z, 12Z octadecadienoic acid; octadecadienoic isomer and 15,16 dihydroxy 9Z, 12Z octadecadienoic acid, 15S, 16S. The 18-carbon fatty acid structure was confirmed by nuclear magnetic resonance (NMR) spectral data. The IC₅₀ value of compounds in Fraction A for anti-inflammatory activity in RAW 264.7 cell line was 434.8±0.75 µg/mL. From the analysis, it can be concluded that Fraction A can be classified under 18 carbon long chain fatty acid group based on LC MS/MS and NMR analysis. This active compound shows an inhibition towards NO activity.

The effect of different concentrations of indoxacarb were studied on adult and embyros of zebrafish (Danio rerio). Acute and chronic toxicity tests were performed on adult and embryos, at different concentrations. At the highest concentration of 30 ppm, there was significant reduction on the hatching rate of embryos. The lethal concentration (LC50) in this study was 1.44 ppm. In acute and chronic toxicity on adult zebrafish, at concentrations of 0.5 ppm and 1.0 ppm, survival was until 14 days. Based on our findings we suggest to reduce the chemical footprint, if not on a total ban on indoxacarb, especially for agricultural applications.

The effect of different concentrations of indoxacarb were studied on adult and embyros of zebrafish (Danio rerio). Acute and chronic toxicity tests were performed on adult and embryos, at different concentrations. At the highest concentration of 30 ppm, there was significant reduction on the hatching rate of embryos. The lethal concentration (LC50) in this study was 1.44 ppm. In acute and chronic toxicity on adult zebrafish, at concentrations of 0.5 ppm and 1.0 ppm, survival was until 14 days. Based on our findings we suggest to reduce the chemical footprint, if not on a total ban on indoxacarb, especially for agricultural applications.

Legumes have become increasingly in demand due to the rich nutrient compositions and phytochemicals of pulses. However, the seed coats of some legume food products were removed prior consumption causing the food loss its nutritional value. The red kidney bean, RKB (Phaseolus vulgaris) and chickpea, CP (Cicer arietenum) which are the common beans among population were investigated in this study. Seed coats of these beans were analysed for the nutritional composition, phenolic compound and antioxidant properties. Carbohydrate was the major macronutrient in both seed coats. RKB and CP seed coats showed statistically significant composition of moisture, fat, protein and fibre. The caloric value of RKB seed coat (2.63 kcal/g) is higher than CP seed coat (2.29 kcal/g). Nevertheless, CP seed coat is a better source of fibre (27%) than RKB seed coat. Total phenolic content (TPC) of RKB seed coat was 12.14 mg GAE/g, which is much higher than in CP seed coat (0.25 mg GAE/g). Interestingly, the seed coat of RKB has strong antioxidant potency with DPPH assay (IC50 = 105.18 µg/ml) comparable to standard Trolox (IC50 = 96.42 µg/ml), which is much lower than the seed coat of CP (IC50 = 606.12 µg/ml). In addition, the antioxidant activity was highly correlated with TPC content of both seed coats. These properties make the seed coat of both beans are excellent candidates of potent nutraceutical.

Nigella sativa L. seed, also recognized as black cumin is a medicinal plant that possesses miraculous power of healing due to the abilities to cure various ailments. Owing to the oily properties of plant oil, the oil ester is more preferable exhibiting special features such as non-toxic, outstanding moisturizing action and most importantly the absence of oily texture. In this study, Nigella sativa oil-based ferulate ester was synthesized via transesterification of Nigella sativa oil (NSO) and ethyl ferulate (EF) in the presence of immobilized lipase, Lipozyme RM IM from Rhizomucor miehei. A five-level-four-factor central composite rotatable design (CCRD) from response surface methodology (RSM) was applied to study the influence of synthesis parameters: lipase dosage (50-80 mg), temperature (40-60°C), substrate ratio of NSO:EF (2.5:1-5.5:1 w/w) and time (3-7 hr) aiming for the maximum percentage yield of ester. The optimized synthesis conditions were lipase dosage of 67 mg, temperature of 56°C, substrate ratio (NSO:EF) of 4.4:1 w/w and time of 4 hr. The maximum production of ester obtained was 49.87% which is acceptable with the predicted value of 48.82%.

In vitro cell viability tests are usually done using 3-(4,5 dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTS) method. A new spectrofluorometry method was developed using acridine orange (AO) and propidium iodide (PI) using multi-label microplate reader. Nine biogenic amines (BAs) [histamine (HIM), putrescine (PUT), cadaverine (CAD), 2-phenylethylamine (PHM), tyramine (TYM), tryptamine (TPM), spermine (SPM), spermidine (SPD) and agmatine (AGM)] were exposed to RAW 264.7 macrophage in singles at 37°C with 5% carbon dioxide supplementation for 18–24 hours and cell viability was determined using MTS method and AO/PI developed method using dual-spectrofluorometry. Based on MTS assay, SPM and SPD were found to be cytotoxic and it was supported by AO/PI assay. The precedence of disintegration in the nucleus rather than mitochondria upon cell non-viability was also supported by transmission electron microscopy (TEM). The results showed that AO/PI method could be used as an alternative method to determine cytotoxicity besides usual usage in confocal microscopy.

Jatropha curcas is a medicinal plant with many therapeutic properties such as anti-inflammatory, anti-malaria, anti-cancer and antioxidant. The root extract has been shown to possess high anti-inflammatory activity. Previously, the compounds responsible for this activity have not been fully elucidated. Two fractions (Fraction 1 and Fraction 2) obtained from a preparative HPLC of the root extract showed significant anti-inflammatory and cytotoxic activities in RAW 264.7 murine macrophage cells with Fraction 1 giving higher nitric oxide (NO) inhibition compared to Fraction 2 and L-NAME. Further purification steps involving column chromatography, thin layer chromatography and analytical HPLC of Fraction 1 produced two fractions labeled as Fraction A and Fraction B. Both fractions showed anti-inflammatory activity without cytotoxic activity in RAW 264.7 cells. Liquid chromatography tandem mass spectrometry (LC-MS/MS) analysis showed that Fraction A contained a group of 18 carbon fatty acid compounds consisting of 2 oxooctadecanoic acids; 15, 16 dihydroxy 9Z, 12Z octadecadienoic acid; octadecadienoic isomer and 15,16 dihydroxy 9Z, 12Z octadecadienoic acid, 15S, 16S. The 18-carbon fatty acid structure was confirmed by nuclear magnetic resonance (NMR) spectral data. The IC₅₀ value of compounds in Fraction A for anti-inflammatory activity in RAW 264.7 cell line was 434.8±0.75 µg/mL. From the analysis, it can be concluded that Fraction A can be classified under 18 carbon long chain fatty acid group based on LC MS/MS and NMR analysis. This active compound shows an inhibition towards NO activity.

Nephelium lappaceum also known as Rambutan is a locally grown popular tropical fruit grown in these parts of Asia. 2,2-diphenyl-1-picrylhydrazyl (DPPH) model of in vitro free radical scavenging test was performed with leaf extracts. The extract was also tested on wound healing activity using mice excision model. The anti-inflammatory effect was tested in mice using the formalin induced paw licking test model. In the DPPH test, Nephelium lappaceum leaf extract at all concentrations was found to cause significant free radical inhibition (p<0.05). All concentrations of Nephelium lappaceum leaf extract showed significantly (p<0.05) high wound contraction on Day 6. In the animal model of inflammation, all doses of Nephelium lappaceum showed reduced number of paw licking. In conclusion, Nephelium lappaceum leaf extract demonstrated in vitro free radical scavenging, anti-inflammatory and wound healing effects.

Nephelium lappaceum also known as Rambutan is a locally grown popular tropical fruit grown in these parts of Asia. 2,2-diphenyl-1-picrylhydrazyl (DPPH) model of in vitro free radical scavenging test was performed with leaf extracts. The extract was also tested on wound healing activity using mice excision model. The anti-inflammatory effect was tested in mice using the formalin induced paw licking test model. In the DPPH test, Nephelium lappaceum leaf extract at all concentrations was found to cause significant free radical inhibition (p<0.05). All concentrations of Nephelium lappaceum leaf extract showed significantly (p<0.05) high wound contraction on Day 6. In the animal model of inflammation, all doses of Nephelium lappaceum showed reduced number of paw licking. In conclusion, Nephelium lappaceum leaf extract demonstrated in vitro free radical scavenging, anti-inflammatory and wound healing effects.

Exposure to ethanol during critical period of development can cause severe impairments in the central nervous system (CNS). This study was conducted to assess the neurotoxic effects of chronic embryonic exposure to ethanol in the zebrafish, taking into consideration the time dependent effect. Two types of exposure regimen were applied in this study. Withdrawal exposure group received daily exposure starting from gastrulation until hatching, while continuous exposure group received daily exposure from gastrulation until behavioural assessment at 6dpf (days post fertilization). Chronic embryonic exposure to ethanol decreased spontaneous tail coiling at 24hpf (hour post fertilization), heart rate at 48hpf and increased mortality rate at 72hpf. The number of apoptotic cells in the embryos treated with ethanol was significantly increased as compared to the control. We also measured the morphological abnormalities and the most prominent effects can be observed in the treated embryos exposed to 1.50% and 2.00%. The treated embryos showed shorter body length, larger egg yolk, smaller eye diameter and heart edema as compared to the control. Larvae received 0.75% continuous ethanol exposure exhibited decreased swimming activity and increased anxiety related behavior, while withdrawal ethanol exposure showed increased swimming activity and decreased anxiety related behavior as compared to the respective control. Biochemical analysis exhibited that ethanol exposure for both exposure regimens altered proteins, lipids, carbohydrates and nucleic acids of the zebrafish larvae. Our results indicated that time dependent effect of ethanol exposure during development could target the biochemical processes thus leading to induction of apoptosis and neurobehavioral deficits in the zebrafish larvae. Thus it raised our concern about the safe limit of alcohol consumption for pregnant mother especially during critical periods of vulnerability for developing nervous system.

Sunburn, premature skin aging, skin cancers and suppression of the immune system are linked to exposure of the skin to UV light. In recent years, plant extracts are becoming a popular active ingredient in natural sunscreen formulations. In the present study, the ethyl acetate fraction of Zanthoxylum rhetsa bark (commonly called as Indian prickly Ash) was used as an active ingredient in two sunscreen cream formulations (F1 and F2). Primarily, the constituents present in the active fraction were identified using LC–MS/MS analysis. Coumaric acid, benzoic acid, p-hydroxybenzoic acid and its isomers, hesperitin, trihydroxyoctadecenoic acid and columbamine were identified in the ethyl acetate fraction of Z. rhetsa bark extract. The UV protection properties of the formulated creams were evaluated by assessment of parameters such as their SPF values (F1: 3.60 ± 0.28, F2: 6.90 ± 0.57), UVA effectiveness (moderate for both test formulations) and critical wavelengths (F1: 365.4, F2: 360.3). Moreover, the physicochemical and microbial count of the formulated creams was also assessed based on various parameters such as colour, pH, centrifugation, viscosity and microbial load over a storage period of 28 days. Both formulations showed pseudo plastic behaviour and were stable at all conditions except for samples kept at 40 °C. Altogether, these results suggested that the ethyl acetate fraction of Z. rhetsa bark has great potential to reduce exposure to harmful UVA/UVB radiations and may be utilized as an active ingredient in natural sunscreen formulation.

Chronic exposure to mercury (Hg) can lead to cumulative impairments in motor and cognitive functions including alteration in anxiety responses. Although several risk factors have been identified in recent year, little is known about the environmental factors that either due exposure toward low level of inorganic mercury that may led to the developmental disorders. The present study investigated the effects of embryonic exposure of mercury chloride on motor function and anxiety-like behavior. The embryo exposed to 6 different concentrations of HgCl2 (7.5, 15, 30, 100, 125, 250 nM) at 5 hpf until hatching (72 hpf) in a semi-static condition. The mortality rate increased in a dose dependent manner where the chronic embryonic exposure to 100 nM decreased the number of tail coiling, heartbeat, and swimming activity. Aversive stimulus was used to examine the effects of 100 nM interferes with the development of anxiety-related behavior. No elevation in both thigmotaxis and avoidance response of 6 dpf larvae exposed with 100 nM were found. Biochemical analysis showed HgCl2 exposure affects proteins, lipids, carbohydrates and nucleic acids of the zebrafish larvae. These results showed that implication of HgCl2 on locomotor and biochemical defects affects motor performance and anxiety-like responses. Yet, the potential underlying mechanisms these responses need to be further investigated which is crucial to prevent potential hazards on the developing organism due to neurotoxicant exposure.

Chronic exposure to mercury (Hg) can lead to cumulative impairments in motor and cognitive functions including alteration in anxiety responses. Although several risk factors have been identified in recent year, little is known about the environmental factors that either due exposure toward low level of inorganic mercury that may led to the developmental disorders. The present study investigated the effects of embryonic exposure of mercury chloride on motor function and anxiety-like behavior. The embryo exposed to 6 different concentrations of HgCl2 (7.5, 15, 30, 100, 125, 250 nM) at 5 hpf until hatching (72 hpf) in a semi-static condition. The mortality rate increased in a dose dependent manner where the chronic embryonic exposure to 100 nM decreased the number of tail coiling, heartbeat, and swimming activity. Aversive stimulus was used to examine the effects of 100 nM interferes with the development of anxiety-related behavior. No elevation in both thigmotaxis and avoidance response of 6 dpf larvae exposed with 100 nM were found. Biochemical analysis showed HgCl2 exposure affects proteins, lipids, carbohydrates and nucleic acids of the zebrafish larvae. These results showed that implication of HgCl2 on locomotor and biochemical defects affects motor performance and anxiety-like responses. Yet, the potential underlying mechanisms these responses need to be further investigated which is crucial to prevent potential hazards on the developing organism due to neurotoxicant exposure.

Sunburn, premature skin aging, skin cancers and suppression of the immune system are linked to exposure of the skin to UV light. In recent years, plant extracts are becoming a popular active ingredient in natural sunscreen formulations. In the present study, the ethyl acetate fraction of Zanthoxylum rhetsa bark (commonly called as Indian prickly Ash) was used as an active ingredient in two sunscreen cream formulations (F1 and F2). Primarily, the constituents present in the active fraction were identified using LC–MS/MS analysis. Coumaric acid, benzoic acid, p-hydroxybenzoic acid and its isomers, hesperitin, trihydroxyoctadecenoic acid and columbamine were identified in the ethyl acetate fraction of Z. rhetsa bark extract. The UV protection properties of the formulated creams were evaluated by assessment of parameters such as their SPF values (F1: 3.60 ± 0.28, F2: 6.90 ± 0.57), UVA effectiveness (moderate for both test formulations) and critical wavelengths (F1: 365.4, F2: 360.3). Moreover, the physicochemical and microbial count of the formulated creams was also assessed based on various parameters such as colour, pH, centrifugation, viscosity and microbial load over a storage period of 28 days. Both formulations showed pseudo plastic behaviour and were stable at all conditions except for samples kept at 40 °C. Altogether, these results suggested that the ethyl acetate fraction of Z. rhetsa bark has great potential to reduce exposure to harmful UVA/UVB radiations and may be utilized as an active ingredient in natural sunscreen formulation.

Exposure to ethanol during critical period of development can cause severe impairments in the central nervous system (CNS). This study was conducted to assess the neurotoxic effects of chronic embryonic exposure to ethanol in the zebrafish, taking into consideration the time dependent effect. Two types of exposure regimen were applied in this study. Withdrawal exposure group received daily exposure starting from gastrulation until hatching, while continuous exposure group received daily exposure from gastrulation until behavioural assessment at 6dpf (days post fertilization). Chronic embryonic exposure to ethanol decreased spontaneous tail coiling at 24hpf (hour post fertilization), heart rate at 48hpf and increased mortality rate at 72hpf. The number of apoptotic cells in the embryos treated with ethanol was significantly increased as compared to the control. We also measured the morphological abnormalities and the most prominent effects can be observed in the treated embryos exposed to 1.50% and 2.00%. The treated embryos showed shorter body length, larger egg yolk, smaller eye diameter and heart edema as compared to the control. Larvae received 0.75% continuous ethanol exposure exhibited decreased swimming activity and increased anxiety related behavior, while withdrawal ethanol exposure showed increased swimming activity and decreased anxiety related behavior as compared to the respective control. Biochemical analysis exhibited that ethanol exposure for both exposure regimens altered proteins, lipids, carbohydrates and nucleic acids of the zebrafish larvae. Our results indicated that time dependent effect of ethanol exposure during development could target the biochemical processes thus leading to induction of apoptosis and neurobehavioral deficits in the zebrafish larvae. Thus it raised our concern about the safe limit of alcohol consumption for pregnant mother especially during critical periods of vulnerability for developing nervous system.

Microcosm describes a miniature representation of a living or non-living thing. For example, there are miniature buildings, cars, furniture, dolls, mushrooms, flowers and plants. Microcosm bioluminescent mushrooms are miniatures of real mushrooms that produce light which is visible in the dark as a result of a biochemical reaction.

Microcosm describes a miniature representation of a living or non-living thing. For example, there are miniature buildings, cars, furniture, dolls, mushrooms, flowers and plants. Microcosm bioluminescent mushrooms are miniatures of real mushrooms that produce light which is visible in the dark as a result of a biochemical reaction.

Zanthoxylum rhetsa is an aromatic tree, known vernacularly as "Indian Prickly Ash". It has been predominantly used by Indian tribes for the treatment of many infirmities like diabetes, inflammation, rheumatism, toothache and diarrhea. In this study, we identified major volatile constituents present in different solvent fractions of Z. rhetsa bark using GC-MS analysis and isolated two tetrahydrofuran lignans (yangambin and kobusin), a berberine alkaloid (columbamine) and a triterpenoid (lupeol) from the bioactive chloroform fraction. The solvent fractions and purified compounds were tested for their cytotoxic potential against human dermal fibroblasts (HDF) and mouse melanoma (B16-F10) cells, using the MTT assay. All the solvent fractions and purified compounds were found to be non-cytotoxic to HDF cells. However, the chloroform fraction and kobusin exhibited cytotoxic effect against B16-F10 melanoma cells. The presence of bioactive lignans and alkaloids were suggested to be responsible for the cytotoxic property of Z. rhetsa bark against B16-F10 cells.

The use of herbal medicine and traditional healing practices for diagnosis, treatment and prevention of illness and ailment continue to have more awareness among the scientific community due to their safety and also as a source of alternatives to synthetic products. This research assessed the total phenolic compounds and in vitro total antioxidant potentials of water extracts in selected species of Zingiberaceae rhizomes use as spice, drinks and medicine. DPPH and FRAP were used to determine the antioxidant capacity, total flavonoid, phenolic acids and polyphenol contents assays to evaluate the quality of the antioxidant activity and the control was ascorbic acid. The results showed that all extracts contain significant antioxidant activity with Zingiber officinale having the highest activity in all assays. DPPH (222.30mg/TE/g DW), FRAP (98.04mg/TE/g DW), Flavonoid (38.58mg/NGN/g DW) phenolic acid (10.78mg/GAE/g DW) and polyphenols (22.84mg/GAE/g DW). Significant and positive linear correlation were found in DPPH, FRAP and total flavonoid, phenolic acids and polyphenol contents. This study reveals some phytochemicals present in Zingiberaceae species, which might be responsible for their biological activities and reason for it use in folkloric medicine in Southeast Asia.

Poor systemic delivery of curcumin outside the gut due to its rapid metabolism has severely limited its application to many chronic diseases. Previously, our research group synthesized curcumin analogues 2, 6-bis (2, 5-dimethoxybenzylidene) cyclohexanone (BDMC33) that has potent anti-inflammatory activities. Therefore, the aim of this study is to evaluate the effects of curcumin analog (BDMC33) on the activities of drug metabolizing enzymes in Caco-2 cells, which was compared with that of curcumin and 3-(2-Fluoro-benzylidene)-5-(2-fluorocyclohexylmethylene)-piperidin-4-one (EF-24). BDMC-33 was synthesized through the appropriate reaction of the aromatic aldehyde with cyclohexanone, under base catalyzed aldol condensation, at the ratio of ketone: aldehyde (1:2). Activity of drug metabolizing enzymes such as NADPH-cytochrome p450 reductase (CPR), UDP-glucuronosyltransferase (UGT), glutathione-S-transferase (GST) and Sulfotransferase (SULT) in Caco-2 cells were evaluated upon exposure to 50µM of BDMC33, curcumin, and EF-24, separately, for 4 hours. The BDMC33, EF-24, and curcumin treatments did not affect the activities of UGT, GST, SULT, and CPR in respect to their controls (29.45, 27.18, 23.64 and 2.08µmol/mg), respectively, at all periods of incubation. Hence, BDMC33 was able to maintain the activities of both phases I and II drug metabolizing enzymes, and therefore it could be a potential lead, anti-inflammatory agents.

Despite progressive research being done on drug therapy to treat breast cancer, the number of patients succumbing to the disease is still a major issue. Combinatorial treatment using different drugs and herbs to treat cancer patients is of major interest in scientists nowadays. Doxorubicin is one of the most used drugs to treat breast cancer patients. The combination of doxorubicin to other drugs such as tamoxifen has been reported. Nevertheless, the combination of doxorubicin with a natural product-derived agent has not been studied yet. Morinda citrifolia has always been sought out for its remarkable remedies. Damnacanthal, an anthraquinone that can be extracted from the roots of Morinda citrifolia is a promising compound that possesses a variety of biological properties. This study aimed to study the therapeutic effects of damnacanthal in combination with doxorubicin in breast cancer cells. Collectively, the combination of both these molecules enhanced the efficacy of induced cell death in MCF-7 as evidenced by the MTT assay, cell cycle, annexin V and expression of apoptosis-related genes and proteins. The effectiveness of doxorubicin as an anti-cancer drug was increased upon addition of damnacanthal. These results could provide a promising approach to treat breast cancer patients.

Cell culture involves a complex of processes of cell isolation from their natural environment (in vivo) and subsequent growth in a controlled environmental artificial condition (in vitro). Cells from specific tissues or organs are cultured as short term or established cell lines which are widely used for research and diagnosis, most specially in the aspect of viral infection, because pathogenic viral isolation depends on the availability of permissible cell cultures. Cell culture provides the required setting for the detection and identification of numerous pathogens of humans, which is achieved via virus isolation in the cell culture as the “gold standard” for virus discovery. In this review, we summarized the views of researchers on the current role of cell culture technology in the diagnosis of human diseases. The technological advancement of recent years, starting with monoclonal antibody development to molecular techniques, provides an important approach for detecting presence of viral infection. They are also used as a baseline for establishing rapid tests for newly discovered pathogens. A combination of virus isolation in cell culture and molecular methods is still critical in identifying viruses that were previously unrecognized. Therefore, cell culture should be considered as a fundamental procedure in identifying suspected infectious viral agent.

Poor systemic delivery of curcumin outside the gut due to its rapid metabolism has severely limited its application to many chronic diseases. Previously, our research group synthesized curcumin analogues 2, 6-bis (2, 5-dimethoxybenzylidene) cyclohexanone (BDMC33) that has potent anti-inflammatory activities. Therefore, the aim of this study is to evaluate the effects of curcumin analog (BDMC33) on the activities of drug metabolizing enzymes in Caco-2 cells, which was compared with that of curcumin and 3-(2-Fluoro-benzylidene)-5-(2-fluorocyclohexylmethylene)-piperidin-4-one (EF-24). BDMC-33 was synthesized through the appropriate reaction of the aromatic aldehyde with cyclohexanone, under base catalyzed aldol condensation, at the ratio of ketone: aldehyde (1:2). Activity of drug metabolizing enzymes such as NADPH-cytochrome p450 reductase (CPR), UDP-glucuronosyltransferase (UGT), glutathione-S-transferase (GST) and Sulfotransferase (SULT) in Caco-2 cells were evaluated upon exposure to 50µM of BDMC33, curcumin, and EF-24, separately, for 4 hours. The BDMC33, EF-24, and curcumin treatments did not affect the activities of UGT, GST, SULT, and CPR in respect to their controls (29.45, 27.18, 23.64 and 2.08µmol/mg), respectively, at all periods of incubation. Hence, BDMC33 was able to maintain the activities of both phases I and II drug metabolizing enzymes, and therefore it could be a potential lead, anti-inflammatory agents.

Zanthoxylum rhetsa is an aromatic tree, known vernacularly as "Indian Prickly Ash". It has been predominantly used by Indian tribes for the treatment of many infirmities like diabetes, inflammation, rheumatism, toothache and diarrhea. In this study, we identified major volatile constituents present in different solvent fractions of Z. rhetsa bark using GC-MS analysis and isolated two tetrahydrofuran lignans (yangambin and kobusin), a berberine alkaloid (columbamine) and a triterpenoid (lupeol) from the bioactive chloroform fraction. The solvent fractions and purified compounds were tested for their cytotoxic potential against human dermal fibroblasts (HDF) and mouse melanoma (B16-F10) cells, using the MTT assay. All the solvent fractions and purified compounds were found to be non-cytotoxic to HDF cells. However, the chloroform fraction and kobusin exhibited cytotoxic effect against B16-F10 melanoma cells. The presence of bioactive lignans and alkaloids were suggested to be responsible for the cytotoxic property of Z. rhetsa bark against B16-F10 cells.

The use of herbal medicine and traditional healing practices for diagnosis, treatment and prevention of illness and ailment continue to have more awareness among the scientific community due to their safety and also as a source of alternatives to synthetic products. This research assessed the total phenolic compounds and in vitro total antioxidant potentials of water extracts in selected species of Zingiberaceae rhizomes use as spice, drinks and medicine. DPPH and FRAP were used to determine the antioxidant capacity, total flavonoid, phenolic acids and polyphenol contents assays to evaluate the quality of the antioxidant activity and the control was ascorbic acid. The results showed that all extracts contain significant antioxidant activity with Zingiber officinale having the highest activity in all assays. DPPH (222.30mg/TE/g DW), FRAP (98.04mg/TE/g DW), Flavonoid (38.58mg/NGN/g DW) phenolic acid (10.78mg/GAE/g DW) and polyphenols (22.84mg/GAE/g DW). Significant and positive linear correlation were found in DPPH, FRAP and total flavonoid, phenolic acids and polyphenol contents. This study reveals some phytochemicals present in Zingiberaceae species, which might be responsible for their biological activities and reason for it use in folkloric medicine in Southeast Asia.

Despite progressive research being done on drug therapy to treat breast cancer, the number of patients succumbing to the disease is still a major issue. Combinatorial treatment using different drugs and herbs to treat cancer patients is of major interest in scientists nowadays. Doxorubicin is one of the most used drugs to treat breast cancer patients. The combination of doxorubicin to other drugs such as tamoxifen has been reported. Nevertheless, the combination of doxorubicin with a natural product-derived agent has not been studied yet. Morinda citrifolia has always been sought out for its remarkable remedies. Damnacanthal, an anthraquinone that can be extracted from the roots of Morinda citrifolia is a promising compound that possesses a variety of biological properties. This study aimed to study the therapeutic effects of damnacanthal in combination with doxorubicin in breast cancer cells. Collectively, the combination of both these molecules enhanced the efficacy of induced cell death in MCF-7 as evidenced by the MTT assay, cell cycle, annexin V and expression of apoptosis-related genes and proteins. The effectiveness of doxorubicin as an anti-cancer drug was increased upon addition of damnacanthal. These results could provide a promising approach to treat breast cancer patients.