Penghasilan Sel Hibridoma Menggunakan Teknik Elektrofusion

Pelakuran sel-sel somatik untuk menghasilkan sel hibrid adalah teknik yang telah meluas digunakan dalam kebanyakan bidanq biologi sel. Elektrofusion adalah di antara teknik yang digunakan untuk tujuan pelakuran sel. Fenomena pemecahan membran secara berbalik apabila diaruh oleh arus elektrik adalah faktor utama berlakunya pelakuran sel. Parameter yang sesuai seperti kekuatan medan arus elektrik, komposisi media pelakuran, jenis sel dan cara pengendalian perlu dikaji selidik sebelum melakukan proses pelakuran. Kaj ian dihuat untuk memilih beberapa parameter yang sesuai dalam penghasilan sel hibridoma melalui pelakuran sel Sp2/-Ag14 dengan sel limfosit mencit yang diimunisasi dengan Virus Sampar Ayam (NDV) strain AF2240. Kajian kemandirian menunjukkan osmolari ti media pelakuran, kekuatan dan masa pendedahan medan elektrik adalah parameter yang memheri kesan kepada kemandirian sel. Media pelakuran inositol didapati lebih baik sebagai media pelakuran berbanding media pelakuran glukosa berdasarkan kepada peratus penghasilan sel hibridoma dan kemampuan dalam mengekalkan tahap kemandirian sel. Arus ulang alik pada voltan 350 v/cm dan frekuensi 2 MHz didapati sesuai untuk proses penjajaran sel limfosit dan Sp2/0-Ag14. Arus terus pada voltan 3 kV/cm dengan tiga kali denyutan pada tempoh masa 10 mikrosaat bagi setiap denyutan adalah berkesan untuk mengaruh pelakuran sel. Perawatan dengan media hipo-osmolar 75 mOs/kg juga dapat meninggikan kemungkinan untuk ber lakunya pelakuran. Teknik elektrofusion juga didapati sesuai digunakan jika kepekatan sel yang digunakan adalah kurang daripada 106/ml. Tetapi jika sel yang digunakan adalah pada kepekatan tinggi iai tu melebihi 1011ml, teknik PEG adalah lebih sesuai kerana lebih murah dan mudah. Dalam penghasilan antibodi monoklon, teknik elektrofusion telah berjaya menghasilkan beberapa klon sel hibridoma yang merembeskan antibodi monoklon terhadap virus Sampar Ayam (NDV). Pengkelasan antibodi juga telah dibuat dan didapati kebanyakkan klon merembeskan antibodi kelas IgG yang mempunyai rantai ringan jenis lambda. Analisis imunoblot menunjukkan antibodi monoklon bertindakbalas dengan protein antigen subunit F2

Antioxidant and angiotensin converting enzyme (ACE) inhibitory activities of cocoa (Theobroma cacao L.) autolysates.

The present study investigated antioxidant and angiotensin converting enzyme (ACE) inhibitory activities of cocoa autolysates. After removal of cocoa fat, alkaloids and polyphenols, the remaining proteinous powder was autolyzed at pH 3.5 and 5.2. At similar concentrations, autolysates produced at pH 3.5 indicated the highest reducing power and ACE inhibition activity. However, those generated at pH 5.2 showed the highest antioxidant activity based on β-carotene bleaching assay. The results displayed a dose-dependent trend. Based on amino acids composition, slight differences were detected between autolysates, and as it was found, they were rich in hydrophobic amino acids. Qualitative and quantitative tests were applied to assure that the results from the assays were not due to the polyphenols of cocoa autolysates. Based on the results no polyphenols could be detected from cocoa autolysates. It can be indicated that among other useful substances of cocoa, its peptides and amino acids could contribute to its health-promoting properties. Furthermore, these bioactive substances can be exploited into functional foods or used as a source of nutraceuticals

Properties of kojic acid and curcumin: assay on cell B16-F1

Ultra violet (UV) exposure and oxidative stress are casually linked to skin disorders. They can increase melanin synthesis, proliferation of melanocytes, and hyperpigmentation. It is possible that antioxidants or inhibitors may have a beneficial effect on skin health to reduce hyperpigmentation. In the last few years, a huge number of natural herbal extracts have been tested to reduce hyperpigmentation. The objective of this study was to determine and to compare of kojic acid and curcumin properties to viability cell B16-F1. In this study, our data showed that the viability of cell B16-F1 was 63.91% for kojic acid and 64.12% for curcumin at concentration 100 µg/ml. Further investigation assay of antioxidant activities, indicated that IC50 for kojic acid is 63.8 µg/ml and curcumin is 16.05 µg/ml. Based on the data, kojic acid and curcumin have potential antioxidant properties to reduce hyperpigmentation with low toxicity effect in cell B16-F1

Ficus deltoidea: A Potential Alternative Medicine for Diabetes Mellitus

Ficus deltoidea from the Moraceae family has been scientifically proven to reduce hyperglycemia at different prandial states. In this study, we evaluate the mechanisms that underlie antihyperglycemic action of Ficus deltoidea. The results had shown that hot aqueous extract of Ficus deltoidea stimulated insulin secretion significantly with the highest magnitude of stimulation was 7.31-fold (P < 0.001). The insulin secretory actions of the hot aqueous extract involved K+ ATP channel-dependent and K+ ATP-channel-independent pathway. The extract also has the ability to induce the usage of intracellular Ca2+ to trigger insulin release. The ethanolic and methanolic extracts enhanced basal and insulin-mediated glucose uptake into adipocytes cells. The extracts possess either insulin-mimetic or insulin-sensitizing property or combination of both properties during enhancing glucose uptake into such cells. Meanwhile, the hot aqueous and methanolic extracts augmented basal and insulin-stimulated adiponectin secretion from adipocytes cells. From this study, it is suggested that Ficus deltoidea has the potential to be developed as future oral antidiabetic agent

Effects of oil palm (elais guineensis) fruit extracts on glucose uptake activity of muscle, adipose and liver cells

The effect of oil palm (Elaeis guineensis) fruit aqueous extract (OPF) on glucose uptake activity of three different cell lines was investigated. The cell lines were incubated with different concentrations of OPF to evaluate the stimulatory effect of OPF towards glucose uptake activity of L6 myotubes, 3T3F442A adipocytes and Chang liver cell line. The glucose uptake activities of all tested cells were enhanced in the presence of OPF extract (basal condition). Nevertheless in combination of OPF extract and 100 nM insulin, the glucose uptake activity was only significantly enhanced in L6 myotubes and 3T3F442A adipocytes cell lines. The extracts enhanced the glucose uptake into cells through either insulin-mimetic or insulin-sensitizing property or combination of these two properties. It can be suggested that the OPF extract exerts its antihyperglycemic action partly by mediated glucose uptake into the glucose-responsive disposal cells, muscle, adipose and liver

Chemical profile, total phenolic content, DPPH free radical scavenging and α-lucosidase inhibitory activities of cosmos caudatus kunth leaves

Herbs and medicinal plants are major sources of traditional or folk medicines for many countries of the world, including Malaysia. This study evaluated the bioactive potential of the leaf ethanolic extract and solvent fractions of Cosmos caudatus Kunth, in scavenging free radicals and inhibiting the enzyme α-glucosidase. In addition, their metabolite profiles were also characterized using liquid chromatography–mass spectrometry. The bioactivity was found to be concentrated in the EtOAc and BuOH fractions which largely contained rutin, quercetin 3-O-galactoside, quercetin 3-O-glucoside, quercetin 3-O-xyloside, quercetin 3-O-arabinofuranoside, quercetin 3-O-rhamnoside, and quercetin 3-O-galactoside, as profiled by LC-MS/MS. It was further shown that the flavonoids glycosides contributed to the free radical scavenging and glucose lowering effects of C. caudatus leaves. The results indicated that the leaves of C. caudatus are a rich source of bioactive compounds and could be prospective materials for development of new anti-diabetic agents

Insulinotropic activity of standardized methanolic extracts of Ficus deltoidea from seven varities

Ficus deltoidea is a traditional medicinal plant that has been proven to show antidiabetic effects. This study focus is to assess the insulin secretion activity of Ficus deltoidea standardized methanolic extracts from seven independent varieties and mechanisms that underlie the insulin secretion action of the extracts. The cytotoxicity of Ficus deltoidea extracts was tested using viability assay. The insulin secretion assay was carried out by treating clonal BRIN BD11 cell line with standardized methanolic Ficus deltoidea extracts or glybenclamide. The clonal BRIN BD11 cell was also treated with insulin agonist and antagonist to elucidate the insulin secretion mechanism. Only the viability percentage for Ficus deltoidea var. kunstleri and intermedia was identified to be toxic at 500 and 1000 μg/ml (P<0.001). The insulin secretion for Ficus deltoidea var. deltoidea, angustifolia, and motleyana was dose-dependent; further evaluation suggested that Ficus deltoidea var. trengganuensis was involved in K ᴬᵀᴾ -independent pathway. This study suggests that standardized methanolic extracts of Ficus deltoidea varieties have an insulinotropic effect on clonal BRIN BD11 cell line and can be utilized as a modern candidate of antidiabetic agents targeting the escalation for insulin secretion from pancreatic beta cells

Penguasaan Dan Kepemilikan Hak Atas Tanah Menurut Hukum Adat Larvul Ngabal

Customary law develops following the development of society and existing folk traditions. Customary law is a deposit of decency in a society whose truth is recognized in that society. Purposes of the research: 1. To find out and analyze how land rights are regulated based on Larvul Ngabal Customary Law. 2. To find out and analyze the obstacles to enforcing Larvul Ngabal customary law. Methods of the research: Normative Juridical Law is used as a type in this study. This type of research in a normative juridical manner conducts studies originating from legal materials, doctrines, principles, legal principles, as well as positive legal provisions that aim to answer the legal issues being studied. Another name for normative research is doctrinal legal research which is commonly called library research because the purpose of this research is written regulations or other legal materials. The results of the study show that the contents of the Larvul Ngabal Law do not specifically regulate the regulation of land rights, control and ownership of land parcels by the Kei indigenous people, which are regulated by each Orong Kai (Head of Ohoi) and Head of Marga based on the regulations in force within the indigenous peoples. . The regulations for each Ohoi regarding land tenure and ownership existed long before the Larvul Ngbal Law existed. Obstacles in upholding larvul Ngabal customary law, namely with the development of the era the habits and understanding of indigenous peoples faded, making it difficult for customary functionaries to apply customary law, apart from that, there was no support from the local government in this matter to form a regional regulation that supports the implementation of the law. customs related to land rights

Effect of ficus deltoidea extracts on hepatic basal and insulin-stimulated glucose uptake

The present study was designed to evaluate the potential of five extract and three fractions of Ficus deltoidea to enhance basal and insulin-stimulated glucose uptake into Chang liver cell line. The results showed that all Ficus deltoidea extracts and fractions except petroleum ether extract have the ability to enhance either basal or insulin-stimulated glucose uptake into liver cell line. Ethanolic and methanolic extracts as well as acidified chloroform and bacified chloroform fractions possess insulin-mimetic activity. Of all extracts and fractions, ethanolic extract possess the highest insulin-mimetic activity. Methanolic extract and n-butanolic fraction possess insulin-sensitizing activity, with the highest activity shown by methanolic extract. There is no synergistic effect between Ficus deltoidea extracts or fractions with 100 nM insulin. It can be suggested that antidiabetic action of Ficus deltoidea is partly associated with glucose disposal into liver cells

Scopariadulcis (SDF7) endowed with glucose uptake properties on L6 myotubes compared insulin.

Aim of the study Insulin stimulates glucoseuptake and promotes the translocation of glucose transporter 4 (Glut 4) to the plasma membrane on L6myotubes. The aim of this study is to investigate affect of Scopariadulcis Linn water extracts on glucoseuptake activity and the Glut 4 translocation components (i.e., IRS-1, PI 3-kinase, PKB/Akt2, PKC and TC 10) in L6myotubescompared to insulin. Materials and methods Extract from TLC fraction-7 (SDF7) was used in this study. The L6myotubes were treated by various concentrations of SDF7 (1 to 50 μg/ml) and insulin (1 to 100 nM). The glucoseuptake activities of L6myotubes were evaluated using 2-Deoxy-D-glucoseuptake assay in with or without fatty acid-induced medium. The Glut 4 translocation components in SDF7-treated L6myotubes were detected using immunoblotting and quantified by densitometry compared to insulin. Plasma membrane lawn assay and glycogen colorimetry assay were carried out in SDF7- and insulin-treated L6myotubes in this study. Results Here, our data clearly shows that SDF7 possesses glucoseuptakeproperties on L6myotubes that are dose-dependent, time-dependent and plasma membrane Glut 4 expression-dependent. SDF7 successfully stimulates glucoseuptake activity as potent as insulin at a maximum concentration of 50 μg/ml at 480 min on L6myotubes. Furthermore, SDF7 stimulates increased Glut 4 expression and translocation to plasma membranes at equivalent times. Even in the insulin resistance stage (free fatty acids-induced), SDF7-treated L6myotubes were found to be more capable at glucose transport than insulin treatment. Conclusions Thus, we suggested that Scopariadulcis has the potential to be categorized as a hypoglycemic medicinal plant based on its good glucose transport properties