Abstract:The aims of this study were to determine the antioxidant and antiproliferative activity of the following Theobroma cacao plant part methanolic extracts: leaf, bark, husk, fermented and unfermented shell, pith, root, and cherelle. Antioxidant activity was determined using 2,2-diphenyl-2-picrylhydrazyl (DPPH), thiobarbituric acid-reactive substances (TBARS), and Folin-Ciocalteu assays; the 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium (MTT) assay was used to determine antiproliferative activity. The root extract had the highest antioxidant activity; its median effective dose (EC50) was 358.3 ± 7.0 µg/mL and total phenolic content was 22.0 ± 1.1 g GAE/100 g extract as compared to the other methanolic plant part extracts. Only the cherelle extract demonstrated 10.4% ± 1.1% inhibition activity in the lipid peroxidation assay. The MTT assay revealed that the leaf extract had the highest antiproliferative activity against MCF-7 cells [median inhibitory concentration (IC50) = 41.4 ± 3.3 µg/mL]. Given the overall high IC50 for the normal liver cell line WRL-68, this study indicates that T. cacao methanolic extracts have a OPEN ACCESSMolecules 2014, 19 18318 cytotoxic effect in cancer cells, but not in normal cells. Planned future investigations will involve the purification, identification, determination of the mechanisms of action, and molecular assay of T. cacao plant extracts.

While considerable information has been presented recently on the alleviating effects of calcium (Ca) on aluminium (AI) toxicity, the interaction between Ca and AI on nodulation and N2-fixation of legumes is little understood. A 28 d solution culture experiment using groundnut (Arachis hypogaea L.) cv. Matjam was conducted to evaluate the effects of four Ca concentrations and four A1 levels on nodule development, N2-fixation and plant growth. The Ca concentrations were maintained at 500, 1000, 2500 or 5000/xM, and the sum of activities of monomeric AI species (ZaAI . . . . ) were 0, 15, 30 and 60/~M. With ZaAuno,o >I-30 ~M in solution, the time to appearance of the first nodule increased, and, with 60/zM ,~aA~ . . . . in solution, plants remained chlorotic throughout the experiment. Activities~ > 3 0 / x M reduced nodule number and nodule dry mass per plant, particularly with high (5000/xM) Ca in solution. Also, plant top growth was decreased at ~anlmono/> 30 /xM; the effect only being alleviated by 1000/zM Ca at 30/zM 2aA~ . . . . . The Ca concentration in the youngest expanded leaf (YEL) increased with increased Ca concentration in solution, but was little affected by AI treatment. Nitrogen concentrations mirrored treatment effects on nodule number and nodule dry mass; AI in solution decreased the N concentration particularly with 5000 ~M Ca in solution. Furthermore, increased Ca and AI in solution decreased the Mg concentration in the YEL. This suggested that the absence of any alleviating effect of Ca and AI toxicity (indeed the opposite effect was often observed) resulted from interference in Mg nutrition.

Cocoa is a rich source of dietary polyphenol, highly potential antioxidant against free radicals. This study was designed to identify the effect of cocoa polyphenol extract in protecting from ethanol-induced liver injury in rats. Fifty male Sprague-dawley rats were divided into five groups fed with or without ethanol (4 g/kg/d), cocoa extract (300 mg/kg/d) and silymarin (200 mg/kg/d) continuously for 3 weeks using an enteral feeding protocol. All treatments were given orally every day for three weeks and continuously supply food and water ad libitum. Results showed that cocoa extract (CE) from unfermented cocoa beans had a total polyphenol content of 335.70±27.51 mg GAE/g and 38.10±4.52 mg CaE/g. Meanwhile, analysis normal phase-high performance liquid chromatography shows CE contains 59.47±9.44 mg/g and 14.69±1.63 mg/g of epicathechin and catechin, respectively, which is three fold higher compared to commercial cocoa powder. It also contains 59.69±2.15 mg/g theobromine which also three fold higher compared to caffeine 19.87±1.37 mg/g. In vitro study showed cocoa extract contains high antioxidant activities by 91.9±1.00 % against superoxide scavenging system (O 2-) and 97.7±0.15% against α-α-diphenyl-β-picrylhydrazyl radical (DPPH) systems. In vivo study showed increasing level in both liver function enzymes, aspartase aminotransferase (AST) and alanine aminotransferase (ALT) in ethanol intoxication by 116.80±5.23 mmol/L and 56.37±2.71 mmol/L, respectively. Ethanol intoxication was blocked by cocoa extract nearly 89.95±1.18 mmol/L and 46.75±0.74 mmol/L, respectively, and it was comparable with SDT group for both enzymes AST and ALT by 112.19±6.02 mmol/L and 42.49±0.62 mmol/L, respectively. Furthermore, ethanol groups showed significantly lower (p<0.05) of glutathione level by 0.29 ±0.03 µmol/g, however cocoa extract with antioxidant defense system either direct or indirectly protect liver injury by increasing glutathione level at 0.53±0.02 µmol/g. As a result, cocoa extract shows its potential as antioxidant agents to protect ethanol-induced liver injury.

The cocoa pod borer, Conopomorpha cramerella (Snellen) is a serious pest in cocoa plantations in Southeast Asia.  It causes significant losses in the crop.  Unfortunately, genetic resources for this insect is extremely scarce.  To improve these resources, we sequenced the transcriptome of C. cramerella representing the three stages of development, larva, pupa and adult moth using Illumina NovaSeq6000.  Transcriptome assembly was performed by Trinity for all the samples.  A total number of 147,356,088 high quality reads were obtained.  Of these, 285,882 contigs were assembled.  The mean contig size was 374 bp.  Protein coding sequence (CDS) was extracted from the reconstructed transcripts by TransDecoder.  Subsequently, BlastX and InterProScan were applied for homology search to make a prediction of the function of CDS in unigene.  Additionally, we identified a number of genes that are involved in reproduction and development such as genes involved in general function processes in the insect.  Genes found to be involved in reproduction such as porin, dsx, bol and fruitless were associated with sex determination, spermatogenesis and pheromone binding.  Furthermore, transcriptome changes during development were analysed.  There were 2,843 differentially expressed genes (DEG) detected between the larva and pupa samples.  A total of 2,861 DEG were detected between adult and larva stage whereas between adult and pupa stage, 1,953 DEG were found.  In conclusion, the transcriptomes could be a valuable genetic resource for identification of genes in C. cramerella and the study will provide putative targets for RNAi pest control.