Penentuan Aktivitas Antikanker Fraksi Etil Asetat Daun Bandotan (ageratum conyzoides linn.) Terhadap Cell Line Kanker Kolon WiDr

 A research to test the anticancer activity of in vitro anticancer activity against colon cancer cell line WiDr Ageratum conyzoides Linn.leafs. The aims of this study are to determine anticancer activity using MTT method. The method used is the extraction of secondary metabolites using maceration with n-hexane, ethyl acetate and ethanol, followed by TLC. Crude extract maceration results that have the highest yield selected and separated by fractions using KKV. Fractions with the greatest weight was chosen to be tested anticancer activity against WiDr cell line by MTT assay. Phytochemical screening and identification of compounds by GC-MS carried out on the fraction which has the lowest IC50 value. Maceration results showed that crude ethyl acetate extract had the highest amount of yield at 7.31% and has the best separation results of TLC is characterized by the highest number of stains. Separation by KKV produce 21 fractions, 4 fractions were selected based on the weight of the total of the fraction 6, 13, 14 and 15. Test anticancer activity with a concentration of 62.5; 125; 250; 500 and 1000 mg mL-1 shows the fraction 6 has the lowest IC50 value is 251.48 mg mL-1. Based on this, the fraction 6 does not have anticancer activity. Although, the phytochemical screening showed alkaloids and terpenoids as well as the result of identification with GC-MS indicated the presence of compounds 2H-benzopyran and neophytadiene Keywords: Ageratum conyzoides Linn. leaves, Column Cromatography Vacuum, anticancer, phytochemical, GC-M

Identifikasi senyawa-senyawa fraaksi kloroform dari kulit batang.

Sebagai usaba untuk menggali amber daya alam nabati, terutama dari gudut metabolit sekunder, telah dilakukan penelitian terftsdap Artocarpus elasticus, suatu tanaman khas Indonesia yang keberadaannya raasih jarang. Penelitian ini dilakukan untuk mengisolasi senyawa-senyawa dalam ekstrak klorofonn kulit batang species tersebut. Peraisahan dilakukan dengan kromatografi kolom dan kromatografi kolom vakum, berfasa diam silika gel. Setelah melalui proses pemurnian, berhasil diperoleh 2 senyawa, yaitu senyawa T dan senyawa Berdasarkan basil analisis terbadap senyawa basil isolasi dan melalui pendekatan khemotaksonomi, diusulkan bahwa senyawa I merupakan golongan triteipenoid dengan gugus ester. Senyawa II merupakan suatu glikosida, yang setelah melalui hidrolisis dengan a.sam klorida, berhasil dipisahkan antara gula dan aglikon. Analisis lebih lanjut me;nunjukkan bahwa jenis gula yang terikat adalah xilosa, sedangkan aglikonnya memberi basil positif terbadap uji golongan triteipenoid. In an effort to explore the plant potention of natural resources, especially from secondary metabolism's corner, there had been done an investigation to Artocarpus elasticus, which is rare species endemic in Indonesia. The research was done to isolate some chemical constituents in chloroform extract from tree bark of this species. The coloumn chromatography and the vacuum colour= chromatography were choosen as separation methods, using silica gel as a stationary phase. After purifying process, two compounds, compound I and II respectively could be isolated. Base on the chemical analysis of these compounds and through chemotaxonomy approach, the first compound was proposed as triterpene with esther group. The second compound was identified as a glycoside, but it was succesfully to be separated between sugar and aglicone by hydrolysis using chloride acid. Further analysis showed that the sugar bonded at the second compound was xylose, while the aglicone gave positive test on triterpene analysis

Flow cytometric analysis of MCF-7 cell line in its treatment with leaves extract of Eugenia uniflora L

The use of natural product in treatment of cancer has attracted more attention since the existing treatments have not yet given any satisfactory result. Dewandaru (Eugeniauniflora.L), one of promising medicinal plants in Indonesia, was reported to have suppressionactivity on DNA polymerase of EBV (Eipstein-Bar Virus). This study aimed to reveal thecytotoxicity of E.uniflora L leaves extract and examine more deeply whether or not thisactivity will trigger the apoptosis process of human breast cancer MCF-7. The extraction process was conducted using petroleum ether, dichloromethane andmethanol. Each extract was subjected for MTT assay for cytotoxicity analysis.Theprospective compounds were then separated using vacuum column chromatography andpreparative thin layer chromatography.MTT method was used to performed cytotoxicity testof each separated fraction on MCF-7 cells. The selected compound which show the mostpotential activity was analyze to its ICso value. MCF-7 cells whichtreated with this toxiccompound was analyze for its cell cycle using flow cytometry assay and propidium iodide staining. The results showed that the ICso of the toxic compound tested on MCF-7 cells were 10ug/mL.Flow cytometry analysis showed that this compound has capability in inducingapoptosis.Cell cycle arrest was observed in MCF-7 cells in which cell accumulation occurredin Gl phase

Marine Actinomycetes as a Potential Source of Bioactive Compounds

The Sea is a complex world which have diverse forms of life including the existence of extreme variation of pressure, salinity and temperature. Marine actinomycetes growing in different habitats than terrestrial actinomycetes, with different stress and obstacles, and become dependent on the marine environment. Adaptation of marine actinomycetes against this condition, allegedly causing microorganisms have the ability to synthesize themetabolite that is unique and not found on the mainland. Recently, the rate of discovery of new compounds from existing generaobtained from terrestrial sources has decreased. Thus, it is crucial that newgroups of actinomycetes from unexplored or underexploited habitats bepursued as sources of novel compounds. The diversity of metabolitesproduced by marine actinomycetes is a source for the discovery of newbioactive compounds. These efforts can be done through two approaches,namely the molecular detection of genes encoding the enzymes that play arole in the biosynthesis of certain secondary metabolites and the isolation ofactive compounds by bioassay guided isolation that combines chemical separation techniques with in vitro testing techniques. Identification of marine actinomycetes producing bioactive compounds can be conducted by analyzing its 16S rRNA gene sequences as molecular chronometers.Phylogenetic analysis will show a relationship between the isolates that we found with isolates that have been found previously

TOXIC COMPOUNDS EXTRACTED FROMI EUGENIA UNIFLORA LAGAINST T47D CELL LINE

Breast cancer is a type of cancer most commonly had by women and is the second most frequentcause of death after lung and bronchial cancer.Today, cancer treatment using natural products has attractedmore attention as the existing treatments do not provide satisfactory results. Dewandaru(Eugeniauniflora.L),also known as the Surinam Cherry,has been reported to have antioxidant activity and cansuppress DNA polymerase of EBV (Eipstein-Bar Virus); however, research on its anticancer activity has notbeen reported.Therefore,the objective of this research is to study the potential application of E. uniflora asanticancer and isolate the toxic compound on T47D cell line. Petroleum ether,dichlormethane and methanol are used to extract E. uniflora leaves.The extract wastested for its cytotoxicactivity on T47D cell line using MTT method.The toxic compounds were separatedusing vacuum column chromatography and Preparative Thin Layer Chromatography methods.Cytotoxicitytest on T47D cells was performed for extracts of each separation stage.Chemical type identification of thetoxic compounds was performed using TLC, UV and IR spectrophotometry analysis. The results showed that dichlormethane extracts of E. uniflora has cytotoxic potential,having ICsovalue of 96 μg/ml and The ICso values of the compounds was 8 μg/ml.Finally,the isolated compounds arebelieved to be of triterpenoid group

UJIAKTIVITAS EKSTRAK DAUN KATU(Sauropus androgynus L. Merr.) SEBAGAI ANTIOKSIDAN PADA MINYAK KELAPA

Sirih merupakan tanaman yang tumbuh subur di Indonesia dan telah diketahui mengandung berbagai macamakst salah satunya sebagai antioksidan Penelitian ini bertujuan untuk mengetahui aktivitas--aatan ekstrak daun katu dalam menghambat oksidasi minyak kelapa. Sampel yang digunakan adalahm latu(Sauropus androgynus L.Merr.)yang diperoleh dari daerah Purworejo.Penelitian ini dilakukann mengekstraksi senyawa dalam daun katu menggunakan tiga pelarut dengan variasi kepolaran,yaitua,oroform,dan metanol. Uji aktivitas antioksidan dilakukan dengan mengukur banyaknyata minyak kelapa dengan metode Tiosianat dari hari pertama sampai dengan hari keempat oksidasi.Sampel yang digunakan adalah kulit pisang ambon kuning yang diperoleh dari pasar tradisional Gowokkualitatif dilakukan dengan menggunakan metode MicroConway Diffusi sedangkan Analisisaifrya.Dari hasil uji aktivitas antioksidan diketahui bahwa ekstrak n-heksana dan kloroform daun katukomsentrasi 0.05%(v/v) dalam minyak kelapa memiliki aktivitas antioksidan yang lebih kecil daripadaT 105%(v/v) yang merupakan kontrol positif. Sedangkan ekstrak metanol mempunyai aktivitassaticksitan yang lebih besar daripada kontrol positif dengan aktivitas antioksidan optimum sebesar 72% padaman petama dan selanjutnya mengalami penurunan hingga hari keempat. Berdasarkan hasil tersebut dapathmpukan bahwa ekstrak metanol daun katu adalah ekstrak yang memiliki aktivitas antioksidan optimumengan aktivitas antioksidan sebesar 72% pada hari pertama

AKTIVITAS FRAKSI n-HEKSANA DAN ETANOL DAUN SIRIH (Piper bettle Linn.)TERHADAP Staphylococcus aureus, Escherichia coli, dan Bacillus subtilis

Sirih merupakan tanaman yang tumbuh subur di Indonesia dan telah digunakan secara tradisional untuk mengobati berbagai macam penyakit. Penelitian ini bertujuan untuk mengetahui aktivitas antibakteri ekstrak daun sirih terhadap Staphylococcus aureus, Escherichia coli,danBacillus subtilis. Penelitian ini dilakukan dengan mengekstrak senyawa aktif dalam daun sirih menggunakan pelarut n-heksana dan etanol. Identifikasi metabolit sekunder dalam daun sirih dilakukan sesuai prosedur standar untuk skrining fitokimia, sedangkan uji aktivitas antibakteri dilakukan menggunakan metode difusi agar dengan paper disk. Bioautografi dilakukan untuk mengetahui profil metabolit sekunder yang menghambat pertumbuhan bakteri. Sampel yang digunakan adalah kulit pisang ambon kuning yang diperoleh dari pasar tradisional Gowok Analisis kualitatif dilakukan dengan menggunakan metode Micro Conway Diffusi sedangkan Analisis kuantitatifnya. Berdasarkan hasil skrining fitokimia, diketahui bahwa fraksi n-heksana daun sirih mengandung kumarin/antrakuinon dan steroid/terpenoid,sedangkan fraksi etanolnya mengandung senyawa fenol dan kumarin/antrakuinon. Uji aktivitas antibakteri menunjukkan bahwa kedua fraksimampu menghambat pertumbuhan bakteri, tetapi fraksi etanol memilki aktivitas antibakteri yanglebih besar.Nilai MIC (Minimum Inhibitory Concentration) ekstrak n-heksana adalah 6,25 mg/mL,sedangkan ekstrak etanol adalah 1,56 mg/mL

Functional characterization of DYRK1A point mutants related to human monogenic disorders

Functional characterization of DYRK1A point mutants related to human monogenic disorders(Esti Wahyu Widowati)DYRK1A is a member of DYRK (dual-specificity tyrosine (Y) phosphorylation-regulated kinase) family of protein kinases which share conserved structure of the DH (DYRK-homology) box and kinase domain, but differ in their N- and C-terminal sequences. DYRK1A achieves full catalytic activity by tyrosine autophosphorylation which takes place as a one-time event during or immediately after translation. Several truncation mutations, microdeletions and missense variants result in the neurodevelopmental syndrome termed mental retardation autosomal dominant 7 (MRD7). The phenotype of MRD7 includes microcephaly, intellectual disability, epileptic seizures, autism spectrum disorder and language delay and is caused by haploinsufficiency of DYRK1A. This study aims to address missense mutations in kinase domain and provide a functional analysis of those mutants. To characterize the disease-causing missense variants that affect the catalytic domain of DYRK1A, we used a mammalian expression system. Four of the substitutions eliminated tyrosine autophosphorylation (L245R, F308V, S311F, S346P), indicating that these variants lacked kinase activity. Tyrosine phosphorylation of DYRK1A-L295F in mammalian cells was comparable to wild type although this mutant showed lower catalytic activity and reduced thermodynamic stability in cellular thermal assays. One variant (DYRK1A-T588N) with a mutation outside the catalytic domain did not differ from wild type DYRK1A in tyrosine autophosphorylation, catalytic activity or cellular localization. In addition, we investigated two DYRK1A mutants (D138P and K150C) located in the DH-box which correspond to missense variants in the related DYRK1B kinase that are associated with a familial form of metabolic syndrome (AOMS3). The DH-box contributes to the conformational stability of catalytic domain in DYRK1A. Expression in HeLa cells showed that there is no significant difference between DYRK1A-D138P and K150C regarding tyrosine autophosphorylation or catalytic activity. However, reduced tyrosine phosphorylation was observed in both DYRK1A variants when expressed in a bacterial cell free in vitro translation system. In summary, these studies suggest that 1) pathogenic missense variants in the catalytic domain of DYRK1A impair enzymatic function by affecting catalytic amino acid residues or by compromising the structural integrity of the kinase domain and 2) D138 and K150 participate in the maturation of DYRK1A albeit these mutations are compensated under physiological conditions

Senyawa Antikanker Dari Aktinomisetes Laut

Cancer cells show uncontrolled growth and cause inappropriate ratio of alive anddead cell. The selectivity of drugs used in cancer therapy and cancer pathogenicity make itthe second cause of death in the world. Various efforts have been conducted to find suitableanticancer which includes usingmarine micro organisms as bioreactors ofactive metabolites. Among the micro organismscomposing the marine biodiversity, actinomycetes is one of the most interesting groupbecause of their ability to produce various compounds with a wide range of activity,including as anticancer. Strategies to find anticancer compounds from marineactinomycetes can be grouped into two approaches, 1) detection of genes encoding theenzymes that play a role in the biosynthesis of particular secondary metabolites and 2)isolation of active compounds by bioassay guided isolation that combines chemicalseparation techniques with in vitro testing techniques

Revealing The Motifs, Properties, and Phylogeny of Lupeol Synthase Using Bioinformatics Approach

Lupane-type triterpenoid saponins are potent plants’ secondary metabolites for drug development as they showed various activities such as anticancer, Sarcoplasmic Reticulum Ca2+-ATPase (SERCA) activator which is very important for neurons, and antileishmanial. The triterpenoid saponin backbone is produced by cyclization and rearrangement of the 2,3-oxidosqualene precursor by the oxidosqualene cyclase. The type of oxidosqualene cyclase involved determines the type of saponins so it is referred to as a key enzyme. Lupane-type saponins are produced by 2,3-oxidosqualene cyclization through the chair-chair-chair conformation and the formation of various cation intermediates. This study aimed to analyze lupeol synthase, the key enzyme which determines the conversion of 2,3-oxidosqualene into lupane-type saponins.  This in silico project was done using bioinformatics programs including Multiple Em for Motif Elicitation (MEME), ProtParam, and Molecular Evolutionary Genetic Analysis (MEGA-X) for relationship analysis. The amino acid sequences analysis using the MEME program showed that lupeol synthase has QW, DCTAE, and CYCR conserved motifs in the oxidosqualene cyclase family even though some evolutions were also present. Analysis of chemical and physical parameters with ProtParam indicated that lupeol synthase had lower stability than lanosterol synthase from Saccharomyces cerevisiae. The phylogenetic tree showed that lupeol synthase was closely related to other plant oxidosqualene cyclases. The results of this study are expected to support the modification strategy determination to increase the production of lupane-type saponins using a biotechnological approach in the pharmaceutical industry