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Polyphenols extracted from the Green Tea (Camellia sinensis) augments the protective immune responses in mice challanged with Salmonella typhimurium Ratnaningsih, Tri; Asmara, Widya; Sismindari, Sismindari
Medical Journal of Indonesia Vol 13, No 1 (2004): January-March
Publisher : Faculty of Medicine Universitas Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (333.117 KB) | DOI: 10.13181/mji.v13i1.122

Abstract

Green tea is an aqueous infusion of dried unfermented leaves of Camellia sinensis. Numerous biological activities of green tea have been reported. The aqueous infusion and its polyphenolic substance are known for their activity as an antimutagenic,  antibacterial, hypocholesterolemic, antioxidant, and mutagenic of B lymphocyte. Studies have demonstrated that green tea polyphenols increase IL-12 production. Salmonella spp infection is an important public health problem in many countries. Cell-mediated immunity (CMI), especially T-cell help is important for protection against this infection. Recent evidence indicates that IL-12 is one such factor that plays a crucial role in the development of CMI. These studies were carired out to investigate the effect of green tea polyphenols to the immune cellulare in mice responses of mice during Salmonella typhimurium infection. The subject consisted of 36 female mice (Balb/C), 6-8 weeks old, divided into 3 groups. The first group was given 10 mg polyphenols/mouse, the second group was given 5 mg polyphenols/mouse, and the third group as the control. In day 31, all mice were infected with 108 CFU Salmonella typhimurium orally. On day 0, 3, 5, and 7 postinfection, 3 mice from each groups were sacrificed, the splenocytes were extracted and cultured to measure  the level of IFN-g in the supernatan and. The peritoneal macrophages were also extracted and cultured to measure the phagocytic activity. The level of IFN-g in splenocyte culture supernatant  increased during infection  in all groups, but the level of the experimental groups  were higher than in control group. The  percentage of phagocytic activity of peritoneal macrophages were higher in the experimental groups than in the control group. The increase of the phagocytic activities were seen corelate with the level of IFN-g supernatan splenocyte culture. (Med J Indones 2004; 13: 1-7)Keywords: polyphenols, green tea, macrophages, phagocytosis
Cytotoxic effects of methanol extract isolated from Erythrina fusca Lour on cancer cell-lines Sismindari, Sismindari
Journal of the Medical Sciences (Berkala Ilmu Kedokteran) Vol 35, No 2 (2003)
Publisher : Journal of the Medical Sciences (Berkala Ilmu Kedokteran)

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Abstract

Background: Erythrina is a medicinal plant which is frequently used to treat cancer in Africa. People in Java, however, use Erythrina fusca (cangkring) to treat varicella and measles. Previous works demonstrated that the methanol extract of this plants leaves induced DNA topoisomerase II mediated DNA cleavage. This activity has been used widely as a target to find anticancer medicine. In order to be scientifically proofed the activity, therefore, it is necessary to analyze directly on the cancer cell-lines. Objectives: To identify the cytotoxicity effect of methanol extract of E. fusca leaves against cancer cell-lines.Methods: Cytotoxicity analysis of methanol extract isolated from E. fusca leaves was carried out against myeloma and HeLa S-3 cancer cell-lines, and to normal mononuclear cell. The level of cytotoxicity was determined by calculating the level of IC50 which was based on the percentage of the cell death following the 24 hours incubation with the extract.Results: It was demonstrated that this methanol extract was cytotoxic to myeloma and HeLa S-3 cell-lines with the IC50 of 0.005 mg/ml and IC50 of 0.08 mg/ml respectively. The percentage of the cell death on treated normal mononuclear cell with the extract, however, was very much low 110%). This was similar to that on the DMSO treated cells.Conclusion: The methanol extract isolated from E. fusca leaves was demonstrated had a selective cytotoxicity effect, as indicated by the level of the IC50 which was higher to myeloma compared to HeLa S3 cell-line, and had much less cytotoxic on normal mononuclear cells.Key words: Erythrina fusca, cytotoxicity, cancer cell-lines, mononuclear cell
Impact of Curcuma mangga Val. Rhizome Essential Oil to p53, Bcl-2, H-Ras and Caspase-9 expression of Myeloma Cell Line Astuti, Endang; Sunarminingsih, Retno; Jenie, Umar Anggara; Mubarika, Sofia; S, Sismindari
Indonesian Journal of Biotechnology Vol 19, No 1 (2014)
Publisher : Universitas Gadjah Mada

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Abstract

Cancer is a disease, a public health problem, which is found in the world as well as in Indonesia. Ingeneral, some of cancer theraphies are ineffective, characterized by the resistance performance of cancer cell line,the exposed normal cell and by the side effects. Nowadays, studies to fi nd the specifi c and safely anti-cancerdrugs were increased by the time. Several studies revealed that Curcuma mangga Val. Rhizome contains somesecondary metabolites, essential or non-essential oil, which has cytotoxic activities to the cancer cells. Basedon these anti-cancer potentials, this study has several aims to recognize anti-cancer selectivity and molecularmechanism by inducting apoptosis and inhibiting myeloma cell proliferation. To C. mangga Val. essential oil,immunocyto chemical test was performed to determine the expression of p53, caspase-9, Bcl-2, H-Ras proteinwhile TUNEL test was performed to determine the number of apoptosis cells.The results of this study shown that anti-cancer molecular mechanism of C. mangga Val. essential oil tomyeloma cell line was performed by increasing apoptosis; by increasing the expression of pro-apoptosis p53,caspase-9 protein and reducing protein which is increasing proliferation Bcl-2 and H-Ras.
Cytotoxic Activity of Tegari (Dianella nemorosa Lam.) Methanol Extract Against HeLa Cells Karim, Aditya Krishar; ., Sismindari; Asmara, Widya; ., Istriyati
Indonesian Journal of Biotechnology Vol 17, No 1 (2012)
Publisher : Universitas Gadjah Mada

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Abstract

Dianella nemorosa Lam. also known as tegari belonging to the Liliaceae family. This plant has been utilized for Papua traditional medicine as well as anticancer agent. This research examined potential cytotoxic activity of tegari (D. nemorosa) leaves extract against cervical cancer cell line (HeLa). Methanol extract was obtained by extracting the leaves powder using methanol. Extract was then applied into HeLa cell line to find out the cytotoxic activity. MTT [3-(4,5-dimetilthiazol-2-il)2,5-difeniltetrazolium bromida) assay was used to measure the cytotoxic activity. The result indicated that D. nemorosa leaves extract possessed cytotoxic activity in HeLa cell line with IC50 values were 685,69 µg/ml, 506,43 µg/ml and 708 µg/ml at the incubation period of 24, 48 and 72 h respectively. The strongest cytotoxic was showed by methanol extract incubated in 48 h.
POTENSI PENGGUNAAN KITOSAN RANTAI PENDEK SEBAGAI PEMBAWA DALAM PENGHANTARAN GEN; EVALUASI IN VITRO Winarti, Lina; Martien, Ronny; Sismindari, S
STOMATOGNATIC- Jurnal Kedokteran Gigi Vol 10, No 2 (2013)
Publisher : Fakultas Kedokteran Gigi Universitas Jember

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Abstract

Gene therapy involves the introduction of DNA or ribonucleic acid (RNA) into the target cell either to express or suppress the biosynthesis of proteins. The success of gene therapy is highly dependent on a suitable carrier system that can efficiently deliver genes in to specific desired cell with minimum cytotoxicity on target cells. Chitosan is interesting to be used as a gene carrier because it has a high positive charge and low toxicity to cells. Positive charge chitosan can form complexes with the plasmid. DNA complexes provide protection against enzymes degradation and promote internalization of plasmids that have been condensed. In this study the complex formation of chitosan-pEFneo-GFP and chitosan / TPP-pEFneo-GFP is by complex coaservation method. The results of complex analysis with a 0.8% agarose gel electrophoresis for 30 minutes 100Volt showed that the complex was stayed in the well and the plasmids did not migrate like plasmids which were dissolved in the water and buffer solvent. Stability evaluation in storage at room temperature for 14 days showed that the complex with chitosan concentration 0.02%-0.04% were the most stable, so that transfection analysis performed for the complex with chitosan concentration of 0.02%-0.04%. Transfection results showed that chitosan is able to protect plasmid and promote the internalization of plasmid into the nucleus and then expressed as a green luminescence. From these results chitosan potentially be developed as a carrier system in gene delivery.
THE OPTIMISED CONDITIONS OF INDUCTION OF RECOMBINANT RIP rMJC15310 ACTIVITY ISOLATED FROM Mirabilis jalapa L. LEAVES Astuti, Puji; ., Sudjadi; ., Sismindari
INDONESIAN JOURNAL OF PHARMACY Vol 23 No 2, 2012
Publisher : Faculty of Pharmacy Universitas Gadjah Mada, Yogyakarta, Skip Utara, 55281, Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (836.5 KB) | DOI: 10.14499/indonesianjpharm0iss0pp93-98

Abstract

Ribosome Inactivating Proteins (RIPs) are compounds isolated from plants with ability to inhibit protein synthesis. The inhibition of protein synthesis is due to inactivation of ribosomal RNA through a site-specific  deadenylation  mediated  by  RNA  N-glycosidase. Reportedly, RIPs mainly possess wide range of bioactivity including antiviral  activity  against  plant  infections.  Other  activities  of  RIP were  as  abortifacien,  antivirus  and  anticancer.  This  study  was aimed  to  isolate  and  characterize  the  optimum  conditions  for inducing  the  expression  of  recombinant  RIPs  isolated  from  the leaves  of  Mirabilis  Jalapa  L.  We  have  been  successfully  isolated several  RIPs  and  engineered  these  proteins  to  be  expressed  in  E. coli. These recombinant proteins were obtained by screening cDNA library  originated  from  the  mRNA  of  Mirabilis  jalapa  L  leaves,  and inserted  into  pUC19  carrying  lacZ  gene.  The  presence  of recombinant  plasmid  was  tested  by  using  α-complementation assay. Many RIPs have been isolated from plants and these proteins express  enzymatic  activity  by  cutting  supercoiled  double  stranded DNA. One RIP namely rMJC15310 was obtained from this study and the  proteins  having  ~  8kb  in  size,  cut  the  supercoiled  DNA  into linear  form  at  the  concentration  as  low  as  5  µg.  The  ability  to  cut supercoiled  DNA  increased  on  inducing  its  expression  with  0.4% IPTG.Key words:   Ribosome  Inactivating  Proteins  (RIP),  IPTG,  Mirabilis  jalapa L., recombinant protein 
Effects of pH, temperature and storage on the stability of MJ-30 protein isolated from Mirabilis jalapa L leaves ., Sudjadi; Ikawati, Zulies; ., Sismindari; Rahayu, Putu Riana Suastari
INDONESIAN JOURNAL OF PHARMACY Vol 15 No 1, 2004
Publisher : Faculty of Pharmacy Universitas Gadjah Mada, Yogyakarta, Skip Utara, 55281, Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (358.748 KB) | DOI: 10.14499/indonesianjpharm0iss0pp1-6

Abstract

The total protein of Mirabilis jalapa L leaves had the ability to cleave supercoiled DNA and showed toxicity on HeLa and Raji cell-lines. The 30 kD protein (MJ-30) purified by cationic exchange chromatography possessed activites as RIP e.g DNA supercoiled cleavage and RNA N-glycosidase activity. The aim of this study was to prepare pure MJ-30 and to observe the stability of MJ-30 .MJ-30 of M.jalapa L leaves was purified using the combination of ammonium sulphate fractionation and cationic exchange chromatography with NaCl gradient elution. MJ-30 was subjected for its stability to assays against pH, temperature, and storage period.Stability assay showed that MJ-30 is stabil at pH 5-6, then the activity decreased as the pH increased. This protein was stable at 300 – 550C, and the activity decreased when the temperature increased. Storage at 40C, MJ-30 was stable until 12 days but the activity was decreasing. However, at 300C, MJ-30 was stable for 3 days only. Glycerol addition to the MJ-30 solution has made the activity stable for 18 days at 40C and 300C storage.Keyword : Protein MJ-30, Leaves of Mirabilis jalapa L, stability, pH, temperature, storage
EFFECT OF BENZALDEHYDE EXCESS IN THE SYNTHESIS OF LR-2 AND CYTOTOXIC ACTIVITY OF LR-2 AGAINTS HeLa CELL Ritmaleni, Ritmaleni; Arifin, Muhammad Fajar; Laksmiani, Ni Putu Linda; ., Rumiyati; ., Sismindari
INDONESIAN JOURNAL OF PHARMACY Vol 23 No 1, 2012
Publisher : Faculty of Pharmacy Universitas Gadjah Mada, Yogyakarta, Skip Utara, 55281, Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (405.791 KB) | DOI: 10.14499/indonesianjpharm0iss0pp9-17

Abstract

LR-2(4-phenyl-3,4-dihydro-indeno[2’,1’]pyramidine-2(1H)- thione;  Leni  Ritmaleni  2),  which  designed  and  assumed  to  have biologically  activity  as  anticancer,  has  been  successfully synthesized  by  using  the  Biginelli  reaction.  This  research  was aimed  to  investigate  the  effect  of  benzaldehyde  excess  in  the synthesis  of  LR-2  and  to  evaluate  the  cytotoxic  activity  of  LR-2against HeLa cancer cell lines. The synthesis was done by reacting benzaldehyde, 2-indanone and together  with thiourea at one time as  said  as  one  pot  reaction  synthetic  methodology  and  the reaction was acid catalysed. The mole equivalent of benzaldehyde was  in  excess  compare  to  others.  The  effect  of  benzaldehyde  in excess is the higher the mole of benzaldehyde, the lower the yield of  LR-2.  The  cytotoxicity  of  LR-2  was  done  by  using  MTT  method and the LC50 was 268.15 μM.Key words : LR-2, benzaldehyde, cytotoxic, HeLa 
VALIDASI METODE ANALISA PENETAPAN KADAR EPIGALOKATEKIN GALAT DENGAN KROMATOGRAFI CAIR KINERJA TINGGI Sugihartini, Nining; Fudholi, Achmad; Pramono, Suwidjiyo; Sismindari, Sismindari
Pharmaciana Vol 4, No 2 (2014): Pharmaciana
Publisher : Universitas Ahmad Dahlan

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (109.743 KB) | DOI: 10.12928/pharmaciana.v4i2.1567

Abstract

High Performance Liquid Chromatography (HPLC) was one of analytical methodscommonly used to determine the concentration of epigallocatechin gallate (EGCG) on green teaextract. The method must be validated in order to fit to its purpose. The aim of this research was toprove that the used method has selectifity, liniearity, precise, accurate and know limit of detection(LOD) and limit of quantification (LOQ) is acceptable. The selectivity of analytical method wasdetermined by calculating the resolution value between two peak. Data from 10 μg/mL and100 μg/mL with 5 replicates would give precition and accuration. Precition was known from CV value and accuration was known from recovery value in each concentration. Liniearity was knownfrom regression linear between concentration and wide area of peak. From regresion linear couldcalculate LOD and LOQ. Research show that method of analyse have selectificity withRs= 2.27>1.5; liniearity with r= 0.99; precision with CV 8.74% at concentration 200 µg/mL and3.74% at concentration 500 µg/mL; accuration with recovery 99.76% at concentration 200 µg/mLand 100.52% at concentration 500 µg/mL and the value of LOD is 33.28 μg/mL and LOQ is110.93 μg/mL.
Detection of nptII Gene and 35SCaMV Promoter in Tomatoes (Solanum lycopersicum L.) Suratman, A.; Ughude, J. O.; ., Sismindari
Journal of Food and Pharmaceutical Sciences Vol 1, No 1 (2013): J.Food Pharm.Sci (January-April)
Publisher : Fakultas Farmasi, Universitas Gadjah Mada

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (395.879 KB) | DOI: 10.14499/jfps

Abstract

The detection of nptII (kanamycin resistance) as a transgenic marker gene and 35SCaMV as promoter in tomatoes has been carried out. DNA from tomatoes samples was isolated using PureLinkTM plant total DNA purification kit. The purity of DNA samples was estimated using UV-Vis spectrophotometry at 260 nm and 280 nm. They gave an absorbance ratio (A260/A280) of 1.74-1.79 which indicated its purities. The quality of the DNA was confirmed by a clear and thick band, as analyzed in 0.8% agarose gel electrophoresis. In order to identify the transgenic tomatoes, a 786-bp fragment of the nptII gene and a 86-bp fragment of 35SCaMV promoter were amplified using polymerase chain reaction (PCR). PCR reaction was prepared at optimum condition, namely annealing temperature at 56°C and 55°C for nptII gene and 35SCaMV promoter, respectively and 300 ng of DNA template. The PCR results were visualized on 2% agarose gel electrophoresis. The results showed that one of three tomatoes (code ST2) contains 35SCaMV promoter and no tomatoes contain nptII gene, indicating that ST2 is transgenic tomato.Key words: tomatoes, PCR, nptII, 35SCaMV