The Aquilaria malaccensis species of the genus Aquilaria is an abundant source of agarwood resin and many bioactive phytochemicals. Recent data regarding the chemical constituents and biological activities of Aquilaria leaves led us to attempt to qualitatively profile the metabolites of Aquilaria malaccensis leaves from a healthy, noninoculated tree through phytochemical screening, GC-MS, and LC/Q-TOF-MS. The present work is also the first to report the antilipoxygenase activity of A. malaccensis leaves from healthy noninoculated tree and investigate its toxicity on oral mucosal cells. A total of 53 compounds were tentatively identified in the extract, some of which have been described in literature as exhibiting anti-inflammatory activity. A number of compounds were identified for the first time in the extract of A. malaccensis leaf, including quercetin, quercetin-O-hexoside, kaempferol-O-dirhamnoside, isorhamnetin-O-hexoside, syringetin-O-hexoside, myricetin, tetrahydroxyflavanone, hesperetin, sissotrin, and lupeol. The antilipoxygenase assay was used to determine the lipoxygenase (LOX) inhibitory potential of the extract, while a WST-1 assay was conducted to investigate the effect of the extract on oral epithelial cells (OEC). The extract implied moderate anti-LOX activity with IC50 value of 71.6 µg/mL. Meanwhile, the cell viability of OEC ranged between 92.55% (10 µg/mL)–76.06% ± (100 µg/mL) upon treatment, indicating some potential toxicity risks. The results attained encourage future studies of the isolation of bioactive compounds from Aquilaria malaccensis leaves, as well as further investigation on the anti-inflammatory mechanisms and toxicity associated with their use.

Agarwood plant has been used in traditional medicine to treat wide range of disorders aside from the main function of its fragrant wood in perfumery. In this present study, we are interested to maximize the yield of extract and investigate the cytotoxic/anti-cancer effects of uninfected branch extracts. Uninfected branch is readily available from agarwood plantation particularly during the early years before inoculation process that formed the fragrant wood (resin); and as such is seen as an economical raw material. Uninfected branch from Aquilaria subintegra was subjected to response surface methodology (RSM)-guided ethanolic extraction to achieve maximum yield while maintaining the biological activity. Cytotoxicity/anti-cancer assays including cell attachment assay, cell viability assay and sulforhodamine B (SRB) assay were carried out on the extracts. Run 11 with 12 hours extraction time, 50 °C temperature, 100 rpm and 60 ml extraction volume gave the highest yield of 0.2130 ± 0.036 g/g agarwood branch extract (ABE). Experimental extract from run 16 showed the most promising cytotoxic effects against MCF-7 with IC50 of 8 µg/ml. The study showed that ABE possess potential cytotoxic/anti-cancer activities against MCF-7 cells and further research is warranted to identify the bioactive compounds and mechanism of action.

This study aims to identify the major phytochemical constituents in Aquilaria malaccensis (Thymelaeaceae) ethanolic leaf extract (ALEX-M) and elucidate their ability to suppress nitric oxide (NO) production from a murine macrophage-like cell line (RAW 264.7) stimulated by lipopolysaccharide (LPS) and interferon-γ (IFN-γ). Dichloromethane (DCM) and ethyl acetate (EtOAc) fractions of ALEX-M were subjected to column chromatography. Eight known compounds were isolated for the first time from this species. Compounds were identified using spectroscopic techniques (IR, UV, HRESIMS, and 1D and 2D NMR). Anti-inflammatory activity of both extract and isolated compounds were investigated in vitro. The fractions offered the isolation of epifriedelanol ( 1 ), 5-hydroxy-7,4′-dimethoxyflavone ( 2 ), luteolin-7,3′,4′-trimethyl ether ( 3 ), luteolin-7,4′-dimethyl ether ( 4 ), acacetin ( 5 ), aquilarinenside E ( 6 ), iriflophenone-2- O -α- l -rhamnopyranoside ( 7 ), and iriflophenone-3- C -β-glucoside ( 8 ). The findings suggest the pharmacological potential of the crude extract (ALEX-M) and its isolates as natural anti-inflammatory agents, capable of suppressing NO production in RAW 264.7 cells stimulated by LPS/IFN-γ.

The rapid emergence of resistant Gram-negative bacteria and the limited discovery of novel antibiotic is a global healthcare challenge. Many medicinal plants with potent bioactivities have been developed for the treatment of bacterial infections. Aquilaria malaccensis exhibits wide applications from perfumes and aromatic foods ingredients and great potential in medicines. In this study, crude leaf extract of A. malaccensis was evaluated for its antibacterial activity against several pathogenic Gram-negative bacteria. The leaves were processed and extracted by Soxhlet method using ethanol as the solvent. The antibacterial activity of the crude extract was tested by disc diffusion method, minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) against Acinetobacter baumannii (ATCC 19606), Klebsiella pneumoniae (ATCC 10031 and ATCC 700603) and Escherichia coli (ATCC 1129). Using the optimized method, the Soxhlet extract produced a yield of 178.41 mg/g. Treatment of the extract at 200 mg/mL displayed the largest inhibition zones of 14.0 mm and 9.7 mm against A. baumannii and K. pneumoniae ATCC 10031, respectively. In contrast, against E. coli and K. pneumoniae ATCC 700603, smaller zones of inhibitions of 3.3 mm were demonstrated. The MIC values of the extract were 32 mg/mL against A. baumannii and K. pneumoniae ATCC 10031 and 64 mg/mL against E. coli and K. pneumoniae ATCC 700603. The MBC values of the extract were consistent with the MIC values for all the bacteria investigated. Overall, this study was the first to show antibacterial activity of A. malaccensis leaves extract particularly against A. baumannii and K. pneumoniae and potentially develop for the treatment of resistant bacteria.

Aquilaria malaccensis has been traditionally used to treat several medical disorders including inflammation. However, the traditional claims of this plant as an anti-inflammatory agent has not been substantially evaluated using modern scientific techniques. The main objective of this study was to evaluate the anti-inflammatory effect of Aquilaria malacensis leaf extract (ALEX-M) and potentiate its activity through nano-encapsulation. The extract-loaded nanocapsules were fabricated using water-in-oil-in-water ( w/o/w ) emulsion method and characterized via multiple techniques including DLS, TEM, FTIR, and TGA. The toxicity and the anti-inflammatory activity of ALEX-M and the extract-loaded nanocapsules (ALEX-M-PNCs) were evaluated in-vitro on RAW 264.7 macrophages and in-vivo on zebrafish embryos. The nanocapsules demonstrated spherical shape with mean particle diameter of 167.13 ± 1.24 nm, narrow size distribution (PDI = 0.29 ± 0.01), and high encapsulation efficiency (87.36 ± 1.81%). ALEX-M demonstrated high viability at high concentrations in RAW 264.7 cells and zebrafish embryos, however, ALEX-M-PNCs showed relatively higher cytotoxicity. Both free and nanoencapsulated extract expressed anti-inflammatory effects through significant reduction of the pro-inflammatory mediator nitric oxide (NO) production in LPS/IFNγ-stimulated RAW 264.7 macrophages and zebrafish embryos in a concentration-dependent manner. The findings highlight that ALEX-M can be recognized as a potential anti-inflammatory agent, and its anti-inflammatory activity can be potentiated by nano-encapsulation. Further studies are warranted toward investigation of the mechanistic and immunomodulatory roles of ALEX-M.

The Halal food industry is founded based on the concept of Halalan Tayyiban (HT), as commanded in the Quran. However, the current approach of HT, as reflected in the current Halal standards, requirements, and practices, may overlook some components that are supposed to be integral part of the concept. This article utilises the modern food system approach to frame the discussion for a holistic overview that links the cycle of the current food system and the HT concept towards obtaining its conceptual clarity. The attributes of HT in six different components of the established food system – production, distribution, processing, marketing, consumption, and waste recovery; are contextualized. An explicit conceptual clarity of HT shall create a ripple effect towards a change of mindset and behaviour of all stakeholders leading to the attainment of the wholesomeness of HT. Beyond the conceptual clarity, HT attributes can be elegantly positioned in the modern food system as vehicles to provide safe, high quality, good and wholesome food; in an ethical and sustainable ecosystem.

Mammalian cell lines, in particular CHO-K1 is vital for the multibillion dollar biotechnology industry. The majority of large scale bioprocessing of commercially valuable protein biopharmaceuticals is produced using this type of cell. An ideal mammalian cell system as host for biologics production should retain efficient use of energy sources in order to boost productivity at minimum cost. Various analyses such as cell counting and monitoring of specific biochemical responses are used to provide data to enable bioprocess control in order to achieve the ideal system. Our study aimed to see whether global metabolite analysis using Gas Chromatography Mass Spectrometry (GCMS) would be a potential alternative approach in providing data for bioprocess control. In this study, we analyzed metabolites of CHO-K1 cells at different growth phases using GCMS. CHO-K1 cells producing insulin like growth factor-I (IGF1) were obtained from ATCC. Cells were grown in T-flask and incubated at 37°C/ 5% CO2 until 70-80% confluent in RPMI 1640 media. Samples (cells and spent/conditioned media) were taken at designated intervals for routine cell counting (Trypan Blue dye exclusion method); glucose, glutamine and lactate determination (YSI 2700); IGF-1 production (ELISA kit R&D Sstems, Inc); and global metabolite analysis (GCMS). Conditioned media from each time point were spun down before subjecting into GCMS. Data from GCMS was then transferred to SIMCA P+12.0 for chemometric evaluation using Principal Component Analysis (PCA). The first component, PC1 results was able to explain 36% of the variation of the data with clear separation between exponential phase and other phases (initial and death phase). This suggests that GCMS-based global metabolite analysis has the ability to capture cell growth behaviour and offered insights of factors that may influence the biological system.ABSTRAK: Produk yang berupa sel kekal mamalia, terutamnya CHO-K1 adalah penting dan menguntungkan industri bioteknologi. Majoritinya pemprosesan protein biofarmaseutikal secara besar-besaran dihasilkan dengan menggunakan sel jenis ini. Sistem sel mamalia yang ideal sebagai hos untuk penghasilan produk ubatan harus mengekalkan penggunaan sumber tenaga secara efisien untuk meningkatkan produktiviti pada kos yang minima. Pelbagai analisa seperti penghitungan sel dan pemerhatian tindak balas biokimia tertentu digunakan untuk memberikan data dan untuk menentukan bioproses terkawal untuk mendapat sistem yang ideal. Kajian ini dijalankan untuk mengkaji sama ada analisa metabolit global menggunakan Spektrometri Jisim Kromatografi Gas (Gas Chromatography Mass Spectrometry (GCMS)) boleh berpotensi sebagai pendekatan alternatif dalam membekalkan data untuk kawalan bioproses. Dalam kajian ini, metabolit sel CHO-K1 dikaji pada peringkat tumbesaran berbeza menggunakan GCMS. Sel CHO-K1 menghasilkan insulin seperti faktor pertumbuhan-I (IGF1) didapati daripada ATCC. Sel dibesarkan dalam kelalang-T dan dieramkan pada 37°C/ 5% CO2 sehingga 70-80% konfluen dalam perantara RPMI 1640. Sampel (sel dan perantara yang digunakan/dilazimkan) diambil kiraan pada selang masa yang telah ditetapkan untuk sel rutin (kaedah eksklusi pencelup Trypan Blue), glukosa, glutamin dan penentuan laktat (YSI 2700); penghasilan IGF -1 (kit ELISA R&D Sstems, Inc); dan analisa global metabolit (GCMS). Perantara yang dilazimkan dari setiap poin masa dikurangkan kelajuannya pada setiap pusingan sebelum dianalisa secara GCMS. Data daripada GCMS kemudiannya dipindahkan ke SIMCA P+12.0 untuk taksiran kemometrik menggunakan Analisis Komponen Utama (Principal Component Analysis (PCA)). Keputusan komponen pertama, PC1 berupaya menjelaskan variasi 36% data dengan pengasingan jelas antara fasa eksponen dan fasa-fasa lain (fasa permulaan dan fasa akhir). Ini menunjukkan bahawa analisa metabolit global berasaskan GCMS mampu menjelaskan tingkah laku pertumbuhan sel dan memberikan pemahaman tentang faktor-faktor yang mempengaruhi sistem biologikal.KEY WORDS: CHO-K1 cells, IGF-1, metabolomics, metabolite profile, PCA, GCMS.

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