In vitro and in vivo cytotoxic effects of chlorella against various types of cancer

Chlorella is one of the microalgae that had been studied intensively owing to its rapid growth and easily cultured at a large scale compared to other microalgae and valuable nutrient compositions. Numerous studies have observed that Chlorella possess various health benefit including antioxidant, anti-cholesterol, anti-inflammatory, and antiproliferative effect against many types of cancer. In this review paper, the effects of various Chlorella species against cancer cells and animal induced cancer are discussed and an overview on Chlorella is briefed. The Chlorella deleterious effect on cancer through various mechanisms such as enhancement of immune system and apoptosis; improving lipid peroxidation; synthesis and expression of the protein-degrading matrix; and preventing the formation of new blood vessels are elaborated as well. Based on the findings of many studies reported in this article, it can be suggested that Chlorella has the potential in supporting cancer therapy and may develop to become an anti-cancer agent

A-002 (Varespladib), a phospholipase A2 inhibitor, reduces atherosclerosis in guinea pigs

<p>Abstract</p> <p>Background</p> <p>The association of elevated serum levels of secretory phospholipase A₂(sPLA₂) in patients with cardiovascular disease and their presence in atherosclerotic lesions suggest the participation of sPLA₂enzymes in this disease. The presence of more advanced atherosclerotic lesions in mice that overexpress sPLA₂enzymes suggest their involvement in the atherosclerotic process. Therefore, the sPLA₂family of enzymes could provide reasonable targets for the prevention and treatment of atherosclerosis. Thus, A-002 (varespladib), an inhibitor of sPLA₂enzymes, is proposed to modulate the development of atherosclerosis.</p> <p>Methods</p> <p>Twenty-four guinea pigs were fed a high saturated fat, high cholesterol diet (0.25%) for twelve weeks. Animals were treated daily with A-002 (n = 12) or vehicle (10% aqueous acacia; n = 12) by oral gavage. After twelve weeks, animals were sacrificed and plasma, heart and aorta were collected. Plasma lipids were measured by enzymatic methods, lipoprotein particles size by nuclear magnetic resonance, aortic cytokines by a colorimetric method, and aortic sinus by histological analyses.</p> <p>Results</p> <p>Plasma total cholesterol, HDL cholesterol and triglycerides were not different among groups. However, the levels of inflammatory cytokines interleukin (IL)-10, IL-12 and granulocyte-macrophage colony-stimulating factor (GM-CSF) were significantly reduced in the treatment group. This group also had a significant 27% reduction in cholesterol accumulation in aorta compared with placebo group. Morphological analysis of aortic sinus revealed that the group treated with A-002 reduced atherosclerotic lesions by 24%.</p> <p>Conclusion</p> <p>The use of A-002 may have a beneficial effect in preventing diet-induced atherosclerosis in guinea pigs.</p

FILTRATION OF NUCLEOCAPSID PROTEIN OF NEWCASTLE DISEASE VIRUS FOR IMMUNODIAGNOSTIC KIT

The aim of the present study is to purify recombinant nucleocapsid (NP) protein of Newcastle disease virus (NDV) using microfitration fitration system and improve the processing conditions as well. A microporous membrane screening using two different sizes of membrane which are 0.1µm and 0.45µm was performed before further improvement on NP protein filtration was carried out. Several conditions of the independent variables that affect the microfiltration process such as the temperature, transmembrane pressure (TMP), and viscosity were observed. It turned out to be that 0.45µm membrane gave higher yield of NP protein than that of 0.1µm membrane. Thus, 0.45µm membrane was used to improve the filtration process for NP protein. Based on the Full Factorial Design (STATISTICA 8.0, Statsoft, Inc.), eight sets of experiment were designed to identify the best conditions for the NP protein filtration. From these experiments, the optimal conditions that gave the highest yield of NP protein are; TMP 4.5psi, viscosity 2.39cP at 4ºC. Based on ANOVA (analysis of variance), all the independent variables studied do not significantly affect the purification of NP protein. However, based on the lowest p value for each variable corresponding to the response, temperature has the largest effect for the NP protein yield

Determination of IGF-1-Producing CHO-K1 Growth Phases Using GCMS-Based Global Metabolite Analysis

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

Metabolomics profiling of extracellular metabolites in CHO-K1 cells cultured in different types of growth media

An efficient mammalian cell system for producing bioproducts should retain high cell viability and efficient use of energy sources rendering the need to understand the effects of various variables on the cell system. In this study, global metabolite (metabolomics) analysis approach was used to try and understand the relationships between types of media used, culture growth behavior and productivity. CHO-KI cells producing IGF-1 were obtained from ATCC and grown in T-flask (37 °C, 5 % CO(2)) until 70–80 % confluent in RPMI 1640 and Ham’s F12, respectively. Samples were taken at 8-hourly intervals for routine cell counting, biochemical responses, insulin like growth factor—1 (IGF-1) protein concentration and global metabolite analysis (gas chromatography mass spectrometry, GCMS). Conditioned media from each time point were spun down before injection into GCMS. Data from GCMS were then transferred to SIMCA-P + Version 12 for chemometric evaluation using principal component analysis. The results showed that while routine analysis gave only subtle differences between the media, global metabolite analysis was able to clearly separate the culture based on growth media with growth phases as confounding factor. Different types of media also appeared to affect IGF-1 production. Asparagine was found to be indicative of healthiness of cells and production of high IGF-1. Meanwhile identification of ornithine and lysine in death phase was found to be associated with apoptosis and oversupplied nutrient respectively. Using the biomarkers revealed in the study, several bioprocessing strategies including medium improvement and in-time downstream processing can be potentially implemented to achieve efficient CHO culture system

Hypolipidemic Effect of Celastrus paniculatus in Experimentally Induced Hypercholesterolemic Wistar Rats

The objective of the present study was to evaluate hypolipidemic effect of methanolic extract of Celastrus paniculatus in experimentally induced hypercholesterolemic rats. Hypercholesterolemia was induced by feeding the animals with high fat diet. Oral administration of methanolic seed extract (50%) of Celastrus paniculatus at the optimized dose of 65 mg/kg body weight, substantially reduced the plasma total cholesterol, triglycerides and LDL cholesterol in comparison with induced hypercholesterolemic animal group and the results were comparable with the standard hypocholesterolemic drug and almost similar to the control group. Atherogenic index and liver weight of treated animals also showed significant decrease compared to the hypercholesterolemic animals. It substantially increased the HDL cholesterol level as compared to control group. A significant increase in the activities of lipoprotein lipase and plasma LCAT enhanced hepatic bile acid synthesis and thereby, increased degradation of cholesterol to neutral sterols. Furthermore, the activities of HMG-CoA reductase, glucose 6-phosphate dehydrogenase and malate dehydrogenase were significantly reduced. Histological studies showed less cholesterol deposits in the aorta of animals fed with seed extract of C. paniculatus compared to the induced hypercholesterolemic animals not given C. paniculatus supplement