Zerumbone-loaded nanostructured lipid carrier induces apoptosis in human colorectal adenocarcinoma (Caco2) cell line

The incorporation of zerumbone (Zer) into nanostructured lipid carrier (NLC) is hypothesized to increase the efficacy of the drug. Nanostructured lipid carrier has sustained-drug release characteristics and is able to improve the solubility and bioavailability of the lipophilic drug. In this study, the anti-cancer effect of Zer was tested on human colorectal adenocarcinoma (Caco-2) cell line. The effect of Zer, zerumbone-loaded nanostructured lipid carrier (Zer-NLC) and NLC on the Caco-2 cell viability were determined using the MTT assay. The treatment concentration ranges from 0 to 120 μM at four different time intervals (i.e., 0 h, 24 hrs, 48 hrs and 72 hrs) were evaluated. At 24 hrs, the half-growth inhibitory concentration (GI50) of Zer-NLC (i.e., 4.25 μM) is lower than that of Zer (i.e., 23.75 μM). However, Zer outperformed the Zer-NLC at the subsequent time points. Similar trend was observed in other parameters including the cytostatic concentration (CC) and half-lethal concentration 50 (LC50). Phase contrast imaging and AO/PI fluorescence staining were performed at the CC and LC50 values. The morphological changes and the apoptosis features could be seen in cells treated with Zer and Zer-NLC while cells treated with NLC showed minor morphological changes. The cells treated with Zer-NLC demonstrated a slightly slower progression of apoptosis, which could be due to the controlled release of Zer from the NLC matrix. It was concluded that the incorporation of Zer into NLC did not compromise the potency and efficacy of the drug

Extractive purification of recombinant thermostable lipase from fermentation broth of Escherichia coli using an aqueous polyethylene glycol impregnated resin system

This study aimed at recovery of thermostable lipase from Escherichia coli BL21 using porous glass beads grafted with polyethylene glycol (PEG) in aqueous impregnated resins system (AIRS). The influencing parameters such as concentration and pH of extraction solution, concentration of NaCl, size of the beads, and pH of the desorption solution on the partition behaviour of lipase were evaluated. Smaller adsorbent (4 mm) had a 65.5% of recovery yield with approximately two-fold higher purification factor compared to that obtained with the larger adsorbent. Recombinant lipase was purified successfully using AIRS with a purification factor of 7.6 and yield of 78.4% under optimum conditions of 18% (w/w) PEG 4000, 10% (w/w) of potassium citrate at pH 9 with 3% (w/w) of NaCl. Optimum desorption was obtained with 4.0 mm of porous glass beads at pH 9

Enrichment of minor components from crude palm oil and palm-pressed mesocarp fibre oil via sequential adsorption-desorption strategy

Crude palm oil (CPO) and palm-pressed mesocarp fibre oil (PPMFO) consist of minor components such as Vitamin E, phytosterols, squalene, and carotene that play numerous health function in human. However, limitations are present in current separation processes to separate or enrich minor components from plant sources. In this study, interest is focusing on the enrichment efficiency of the minor components from CPO and PPMFO through proposed sequential adsorption-desorption strategy. The strategy started with the adsorption-desorption properties evaluation of six adsorbents. Both Diaion HP20 and Sepabeads SP850 showed better adsorption-desorption properties than silica gel, Florisil, Diaion HP2MG and Amberlite XAD-7HP. Diaion HP20 was selected as the suitable adsorbent to perform a series of selective desorption process using three different solvents: methanol, IPA, and n-hexane in Soxhlet extraction. Vitamin E, phytosterols, and squalene from CPO were obtained in the 1st fraction using methanol solution and their concentration increased from the initial concentration with the enrichment factor (EF) of 3.4, 3.9, and 1.8, respectively, which slightly higher than those minor components obtained from PPMFO, 1.2, 1.8, and 1.4, respectively. The carotene from both CPO and PPMFO was enriched in the 3rd fraction by using n-hexane solution with an enrichment factor of 1.1 and 1.5, respectively. In conclusion, the obtained result revealed the efficiency of the proposed sequential adsorption-desorption strategy to enrich the minor components from CPO and PPMFO

Primary capture of cyclodextrin glycosyltransferase derived from Bacillus cereus by aqueous two phase system.

In this works, the polymer–salt aqueous two phase system (ATPS) which is polyethylene–glycol (PEG) with sodium citrate was constructed to purify the enzyme CGTase from fermentation broth. Impacts of parameters such as phase composition, tie-line length (TLL), volume ratio (VR), crude sample loading, pH and the addition of sodium chloride (NaCl) on the partition behavior of cyclodextrin glycosyltransferase (CGTase) were investigated. The study exhibited that the optimum system for the purification of the enzyme CGTase was achieved on the 19.0% PEG and 11.5% citrate system with TLL of 38.89% (w/w), VR of 2.0, with additional 4% (w/w) NaCl and 20% crude load at pH 7.0. CGTase from Bacillus sp. was partially purified by the ATPS up to 16.3-fold with a yield of 70%

Extractive bioconversion of cyclodextrins by Bacillus cereus cyclodextrin glycosyltransferase in aqueous two-phase system

An extractive bioconversion with Bacillus cereus cyclodextrin glycosyltransferase (CGTase, EC 2.4.1.19) in aqueous two-phase system (ATPS) was investigated for the synthesis and recovery of cyclodextrins (CDs). Optimum condition for the extractive bioconversion of CDs was achieved in ATPS consisted of 7.7% (w/w) polyethylene glycol (PEG) 20,000 and 10.3% (w/w) dextran T500 with volume ratio (VR) of 4.0. Enzymatic conversion of starch occurred mainly in dextran-rich bottom phase whereas the product, CDs was transferred to top phase and a higher partition coefficient of CDs was achieved. Repetitive batch of CDs synthesis was employed by replenishment of the top phase components and addition of starch every 8 h. An average total CDs concentration of 13.7 mg/mL, (4.77 mg/mLα-CD, 5.02 mg/mLβ-CD and 3.91 mg/mLγ-CD) was recovered in the top phase of PEG 20,000/dextran T500 ATPS. This study showed the effectiveness of ATPS application in extractive bioconversion of CDs synthesis with B. cereus CGTase

Optimization of recovery of esterase from Serratia marcescens using combination of the solvent impregnated resin and aqueous two-phase extraction techniques

The performance of tunable aqueous polymer phase impregnated resins (TAPPIR) which is the combination of the solvent impregnated resin principle and an aqueous two-phase system for the separation of esterase from Serratia marcescens was evaluated in this study. Different molecular weight of polyethylene glycol (PEG) (2000, 4000 and 6000) at concentration ranging from 5% to 20% (w/w) and potassium citrate were used to construct the aqueous phase in TAPPIR technology. Optimum composition of PEG and salt for esterase partitioning was determined using response surface methodology. The optimum condition for the purification of esterase was impregnation of 25% (w/w) of PEG 2000 into 4 mm porous glass beads and extraction of esterase using 15% (w/w) potassium citrate at pH 8 containing 12% (w/w) crude loading with the addition of 4% (w/w) NaCl. Esterase from S. marcescens was successfully purified by the TAPPIR technology up to 5.32 of purification factor with a yield of 75.98%

Development of a novel direct compressible co-processed excipient and its application for formulation of Mirtazapine orally disintegrating tablets

Introduction: Orally disintegrating tablets (ODTs) are designed to dissolve in the oral cavity within 3 min, providing a convenient option for patients as they can be taken without water. Direct compression is the most common method used for ODTs formulations. However, the availability of single composite excipients with desirable characteristics such as good compressibility, fast disintegration, and a good mouth-feel suitable for direct compression is limited. Objective: This research was proposed to develop a co-processed excipient composed of xylitol, mannitol, and microcrystalline cellulose for the formulation of ODTs. Methods: A total of 11 formulations of co-processed excipients with different ratios of ingredients were prepared, which were then compressed into ODTs, and their characteristics were thoroughly examined. The primary focus was on evaluating the disintegration time and hardness of the tablets, as these factors are important in ensuring the ODTs meet the desired criteria. The model drug, Mirtazapine was then incorporated into the chosen optimized formulation. Results: The results showed that the formulation comprised of 10% xylitol, 10% mannitol and 80% micro-crystalline cellulose demonstrated the fastest disintegration time (1.77 ± 0.119 min) and sufficient hardness (3.521 ± 0.143 kg) compared to the other formulations. Furthermore, the drug was uniformly distributed within the tablets and fully released within 15 min. Conclusion: Therefore, the developed co-processed excipients show great potential in enhancing the functionalities of ODTs, offering a promising solution to improve the overall performance and usability of ODTs in various therapeutic applications

Drying characteristics of Orthosiphon stamineus Benth by solar assisted heat pump drying

Processing methods of Misai Kucing still remain crude and lack technological advancements. In terms of drying, very few studies have attempted to apply advanced drying technology to improve Misai Kucing quality and drying time. This paper presents first attempt to improve Misai Kucing drying kinetics and product quality through solar-assisted heat pump drying and comparison was made against solar drying. Experimental results showed that solar-dried samples had the greatest total color change and loss of two bioactive ingredients as compared to solar-assisted heat pump-dried samples due to its longer time process, higher drying temperature, and chlorophyll degradation. By comparing the statistical values, it showed that the Page model had the best goodness of fit at all tested dried samples by both drying methods

Study on retention of metabolites composition in misai kucing (orthosiphon stamineus) by heat pump assisted solar drying

In this research, the effects of different drying methods on the two biomarker compounds concentrations, such as sinensetin (SEN) and rosmarinic acid (RA) of Orthosiphon staminues Benth leaves, flowers, stems and mix were investigated. SEN and RA retention among the dehydrated samples were maximum 83.07% in leaves and 92.54% in flowers, respectively by heat pump assisted solar drying method. For heat pump assisted solar dried samples, there are no significant differences (p > .05) in the retention of SEN and RA as compared to fresh samples. However, there are significant differences (p < 0.05) in SEN and RA retention in solar dried leaves, flowers, stems and mix as compared to fresh samples. Besides, an optimization study was performed to evaluate the optimal drying conditions for obtaining high SEN yields from Misai Kucing's leaves, flowers, stems and mixes with high RA contents within the drying parameters

Basic science232. Certolizumab pegol prevents pro-inflammatory alterations in endothelial cell function

Background: Cardiovascular disease is a major comorbidity of rheumatoid arthritis (RA) and a leading cause of death. Chronic systemic inflammation involving tumour necrosis factor alpha (TNF) could contribute to endothelial activation and atherogenesis. A number of anti-TNF therapies are in current use for the treatment of RA, including certolizumab pegol (CZP), (Cimzia ®; UCB, Belgium). Anti-TNF therapy has been associated with reduced clinical cardiovascular disease risk and ameliorated vascular function in RA patients. However, the specific effects of TNF inhibitors on endothelial cell function are largely unknown. Our aim was to investigate the mechanisms underpinning CZP effects on TNF-activated human endothelial cells. Methods: Human aortic endothelial cells (HAoECs) were cultured in vitro and exposed to a) TNF alone, b) TNF plus CZP, or c) neither agent. Microarray analysis was used to examine the transcriptional profile of cells treated for 6 hrs and quantitative polymerase chain reaction (qPCR) analysed gene expression at 1, 3, 6 and 24 hrs. NF-κB localization and IκB degradation were investigated using immunocytochemistry, high content analysis and western blotting. Flow cytometry was conducted to detect microparticle release from HAoECs. Results: Transcriptional profiling revealed that while TNF alone had strong effects on endothelial gene expression, TNF and CZP in combination produced a global gene expression pattern similar to untreated control. The two most highly up-regulated genes in response to TNF treatment were adhesion molecules E-selectin and VCAM-1 (q 0.2 compared to control; p > 0.05 compared to TNF alone). The NF-κB pathway was confirmed as a downstream target of TNF-induced HAoEC activation, via nuclear translocation of NF-κB and degradation of IκB, effects which were abolished by treatment with CZP. In addition, flow cytometry detected an increased production of endothelial microparticles in TNF-activated HAoECs, which was prevented by treatment with CZP. Conclusions: We have found at a cellular level that a clinically available TNF inhibitor, CZP reduces the expression of adhesion molecule expression, and prevents TNF-induced activation of the NF-κB pathway. Furthermore, CZP prevents the production of microparticles by activated endothelial cells. This could be central to the prevention of inflammatory environments underlying these conditions and measurement of microparticles has potential as a novel prognostic marker for future cardiovascular events in this patient group. Disclosure statement: Y.A. received a research grant from UCB. I.B. received a research grant from UCB. S.H. received a research grant from UCB. All other authors have declared no conflicts of interes