Combining In Silico and In Vitro Studies to Evaluate the Acetylcholinesterase Inhibitory Profile of Different Accessions and the Biomarker Triterpenes of Centella asiatica.

Alzheimer's disease (AD) is a neurodegenerative disease and the most cause of dementia in elderly adults. Acetylcholinesterase (AChE) is an important beneficial target for AD to control cholinergic signaling deficit. Centella asiatica (CA) has proven to be rich with active ingredients for memory enhancement. In the present study, the chemical profiling of three accession extracts of CA namely SECA-K017, SECA-K018, and, SECA-K019 were performed using high-performance liquid chromatography (HPLC). Four biomarker triterpene compounds were detected in all CA accessions. Quantitative analysis reveals that madecassoside was the highest triterpene in all the CA accessions. The biomarker compounds and the ethanolic extracts of three accessions were investigated for their acetylcholinesterase (AChE) inhibitory activity using Ellman's spectrophotometer method. The inhibitory activity of the triterpenes and accession extracts was compared with the standard AChE inhibitor eserine. The results from the in vitro study showed that the triterpene compounds exhibited an AChE inhibitory activity with the half-maximal inhibitory concentration (IC50) values between 15.05 ± 0.05 and 59.13 ± 0.18 µg/mL. Asiatic acid was found to possess strong AChE inhibitory activity followed by madecassic acid. Among the CA accession extracts, SECA-K017 and SECA-K018 demonstrated a moderate AChE inhibitory activity with an IC50 value of 481.5 ± 0.13 and 763.5 ± 0.16 µg/mL, respectively from the in silico docking studies, it is observed that asiatic acid and madecassic acid showed very good interactions with the active sites and fulfilled docking parameters against AChE. The present study suggested that asiatic acid and madecassic acid in the CA accessions could be responsible for the AChE inhibitory action and could be used as markers to guide further studies on CA as potential natural products for the treatment of AD.

Enhancing Trichomonas vaginalis Identification in Cytology Students through Gamification.

Background and Objective: Trichomonas vaginalis is a causative agent of trichomoniasis, a sexually transmitted disease. In the cytology course context, students are taught to identify the cytomorphological characteristics of this organism. However, the limited learning period hinders their ability to recognize this infection effectively. This study intended to evaluate the efficacy of gamification by utilizing a web-based online game to enhance students' capacity to recognize Trichomonas vaginalis infection in cytology. Materials and Methods: The study involved 50 Medical Laboratory Technology students who were randomly assigned to three groups. Group 1 (G1) participants received an interactive web-based online game called CytoUniverse, which comprised three components: a story-based game, a cytomorphology game, and a quiz focusing on Trichomonas vaginalis infection in cytology. Group 2 (G2) participants received the same information from a video lecture. Group 3 (G3) received both the web-based online game and the video lecture. The participants were assessed before the intervention (T1) and after the intervention (T2) to measure the effectiveness of the respective learning methods. IBM SPSS version 28 and GraphPad Prism version 9.0 were used to collect, tabulate, and analyze the data. By using descriptive analysis, the normality of the data was checked. Knowledge score and age were described as mean and standard deviation (SD) for numerical data. On the contrary, the categorical data, such as gender and group categories, were reported as frequencies and percentages. Fisher's exact test, paired t-test, and one-way ANOVA test were used in this study to determine the significance between groups. Results: The study's results indicated a statistically significant improvement (P < 0.05) in knowledge scores at T2 compared to T1 for both G1 and G2 when compared to G3. However, there were no significant differences in knowledge scores between all groups for T1 or T2. Conclusions: In conclusion, gamification through a web-based online game may improve understanding of Trichomonas vaginalis infection. It looks to be a promising strategy for boosting students' knowledge and awareness to recognize Trichomonas vaginalis infection in cytology.

Nanoemulsion of Ethanolic Extract of Centella asiatica (NanoSECA) Ameliorates Learning and Memory Performance by Enhancing Cholinergic Activities, Increasing Antioxidative Levels, and Attenuating oxidative stress markers in adult rats.

BACKGROUND: Centella asiatica (C. asiatica) has long been traditionally used as a memory enhancer. Nanoemulsion of ethanolic extract C. asiatica (NanoSECA) has been developed to improve brain functions. However, the effect of NanoSECA on enhancing memory and cognitive functions remains unexplored. OBJECTIVES: This research aimed to investigate the potential of NanoSECA on cognitive tasks and memory enhancement pathways in a normal adult rat model. METHODS: Thirty male Sprague Dawley rats (7-8 weeks old) were randomly subjected to five groups (n=six per group). Treatment groups were supplemented with NanoSECA and ethanolic extract of C. asiatica (SECA) for 28 days by oral gavages. Different brain sections were isolated, homogenized, and tested for acetylcholinesterase, antioxidants (glutathione and malondialdehyde), and anti-inflammatory agents (nitric oxide, tumour necrosis factor-α, and prostaglandin E2). RESULT: NanoSECA supplementation markedly enhanced the acetylcholine, glutathione levels and reduced a distinct diminution in plasma activities of acetylcholinesterase, malondialdehyde, nitric oxide, prostaglandin E, and tumor necrosis factor-α levels. CONCLUSION: NanoSECA can be used as a memory enhancer through enhanced cholinergic activity, increased antioxidant level, and reduced oxidative stress.

Effect of Storage Temperatures on Physico-Chemicals, Phytochemicals and Antioxidant Properties of Watermelon Juice (Citrullus lanatus).

Watermelon (Citrullus lanatus) consists of high moisture content and is favoured for its juice products. The popular fruit has a tempting taste, sweet aroma and attractive flesh colour. It is enriched with phytochemicals and antioxidant properties that are beneficial to human health. Due to convenience, the majority of individuals are likely to consume watermelon juice. However, little is known about the fruit juice storage and temperatures that may affect its beneficial properties. This study investigated the effect of storage temperature at room temperature, refrigerator cold, refrigerator freeze and freeze-dried, and analyzed the juice physico-chemicals (weight loss, pH, ash, moisture, total soluble solid, browning and turbidity), phytochemicals (total phenolic, total flavonoid, lycopene and ß-carotene) and antioxidant scavenging activities during 9 days of storage. The results showed that watermelon juice was affected by storage temperatures and conditions with significant changes in physico-chemical appearance and decrease in total phytochemical content, thus consequently affecting their antioxidant activities during 9 days of storage. Although fresh watermelon juice can be consumed for its high nutritional values, freeze-drying is the preferable technique to retain its benefits and to delay juice degradation.

Development and Optimization of Nanoemulsion from Ethanolic Extract of Centella asiatica (NanoSECA) Using D-Optimal Mixture Design to Improve Blood-Brain Barrier Permeability.

The evidence on the neuroprotective impact of Centella asiatica (C. asiatica) has been greatly documented in recent years. However, a major obstacle that remains to be overcome is the capacity of the active molecules in C. asiatica to cross the blood-brain barrier (BBB). In this study, we explored the possibilities of using a D-optimal mixture design to fabricate nanoemulsion of C. asiatica (NanoSECA) for better brain bioavailability. The parameters for optimization were the percentage of water (10-80% w/v) and virgin coconut oil (VCO) (10-80% w/v). Nanoemulsions were formulated using a high-pressure homogenization approach and were characterized for their physicochemical properties. The optimal VCO-based nanoemulsion (VBN: F2) conditions were found at 80% (w/v) of water and 10% (w/v) of VCO. Subsequently, viability tests were conducted on neuroblastoma (SH-SY5Y) and macrophage (RAW 264.7) cell lines. NanoSECA was distinguished for its antioxidant, acetylcholinesterase (AChE), anti-inflammatory, and parallel artificial membrane permeability assay (PAMPA) activities in vitro. The NanoSECA has a particle size of 127.833 ± 8.280 nm, zeta potential (ZP) of -24.9 ± 0.011 mV, polydispersity index (PDI) of 0.493 ± 4.681, percentage prediction error (PPE) of -12.02%, and pH of 6.0 ± 0.006 and is also stable under different storage conditions. Cell viability was improved in a dose-dependent manner on SH-SY5Y and RAW 264.7 cell lines. In addition, NanoSECA significantly reduced the AChE activity, suppressing the level of proinflammatory mediators and oxidative stress. Moreover, NanoSECA showed high BBB permeation with a high value of experimental permeability to cross the BBB. Thus, NanoSECA could efficiently potentiate the central nervous system (CNS) therapeutic activities through enhanced penetration of BBB. These nano-delivery systems are crucial to unlock the full potential of C. asiatica for treating numerous CNS disorders.

Histopathological changes in placenta and liver of pregnant rats administered with aqueous extract of Dioscorea hispida var. daemona (Roxb) Prain & Burkill.

Dioscorea hispida var. daemona (Roxb) Prain & Burkill (DH), also known a tropical yam or intoxicating yam is a bitter wild tuber which is consumed as a staple food and traditionally used as a remedy in Malaysia. However, DH is also notorious for its intoxicating effects and there is currently a dearth of study of possible effects of DH on liver and placental tissues and hence its safe consumption warrants in-depth investigation. This study was therefore designed to investigate into the effect of DH on liver and placenta of pregnant rat via histopathological examination. Thirty pregnant Sprague-Dawley rats were randomly divided into five groups consisting of a control (distilled water) and four DH aqueous extract groups (250, 500, 1000 and 2000 mg/kg body weight). The extracts were administered via oral gavage daily throughout the study and animals were sacrificed on day 21. Paraffin-embedded, hematoxylin and eosin stained sections of placenta and liver were examined. Significant changes (p < 0.05) were observed on relative liver and placental weights of animals treated with 2000 mg/kg body weight DH extract. The placental numbers were decreased with the increased of DH extract concentration. Liver histological examination in all treated groups showed that tissues underwent degeneration characterized by hepatocyte swelling, cytoplasmic vacuolation, cytolysis, margination and clumping of nucleus chromatin. Changes of the basal and labyrinth zone were observed in placental tissues in all treated groups. Glycogen cells were reduced with fibrin deposition in the basal zone, while irregular vessel formation was demonstrated in the labyrinth zone. UHPLC-ESI-MS analysis showed the presence four steroidal saponins DH. In conclusion, DH aqueous extract exert hepatotoxicity and adverse effects on the placenta of rats. However, the underlying mechanism and phytochemicals inducing the observed toxicity require further investigation.

Development of Isocratic RP-HPLC Method for Separation and Quantification of L-Citrulline and L-Arginine in Watermelons.

Watermelons (Citrullus lanatus) are known to have sufficient amino acid content. In this study, watermelons grown and consumed in Malaysia were investigated for their amino acid content, L-citrulline and L-arginine, by the isocratic RP-HPLC method. Flesh and rind watermelons were juiced, and freeze-dried samples were used for separation and quantification of L-citrulline and L-arginine. Three different mobile phases, 0.7% H3P04, 0.1% H3P04, and 0.7% H3P04 : ACN (90 : 10), were tested on two different columns using Zorbax Eclipse XDB-C18 and Gemini C18 with a flow rate of 0.5 mL/min and a detection wavelength at 195 nm. Efficient separation with reproducible resolution of L-citrulline and L-arginine was achieved using 0.1% H3P04 on the Gemini C18 column. The method was validated and good linearity of L-citrulline and L-arginine was obtained with R2 = 0.9956, y = 0.1664x + 2.4142 and R2 = 0.9912, y = 0.4100x + 3.4850, respectively. L-citrulline content showed the highest concentration in red watermelon of flesh and rind juice extract (43.81 mg/g and 45.02 mg/g), whereas L-arginine concentration was lower than L-citrulline, ranging from 3.39 to 11.14 mg/g. The isocratic RP-HPLC method with 0.1% H3P04 on the Gemini C18 column proved to be efficient for separation and quantification of L-citrulline and L-arginine in watermelons.

Effects of different tube potentials and iodine concentrations on image enhancement, contrast-to-noise ratio and noise in micro-CT images: a phantom study.

The study aimed to investigate the effects of different tube potentials and concentrations of iodinated contrast media (CM) on the image enhancement, contrast-to-noise ratio (CNR) and noise in micro-computed tomography (µCT) images. A phantom containing of five polyethylene tube was filled with 2 mL of deionized water and iodinated CM (Omnipaque 300 mgI/mL) at four different concentrations: 5, 10, 15, and 20 mol/L, respectively. The phantom was scanned with a µCT machine (SkyScan 1176) using various tube potentials: 40, 50, 60, 70, 80, and 90 kVp, a fixed tube current; 100 µA, and filtration of 0.2 mm aluminum (Al). The percentage difference of image enhancement, CNR and noise of all images, acquired at different kVps and concentrations, were calculated. The image enhancement, CNR and noise curves with respect to tube potential and concentration were plotted and analysed. The highest image enhancement was found at the lowest tube potential of 40 kVp. At this kVp setting, the percentage difference of image enhancement [Hounsfield Unit (HU) of 20 mol/L iodine concentration over HU of deionized water] was 43%. By increasing the tube potential, it resulted with the reduction of HU, where only 17.5% different were noticed for 90 kVp. Across all iodine concentrations (5-20 M), CNR peaked at 80 kVp and then these values showed a slight decreasing pattern, which might be due insufficient tube current compensation. The percentage difference of image noise obtained at 40 and 90 kVp was 72.4%. Lower tube potential setting results in higher image enhancement (HU) in conjunction with increasing concentration of iodinated CM. Overall, the tube potential increment will substantially improve CNR and reduce image noise.