The Dose Effect of Mangrove Leaf Extract (Rhizophora apiculata) on Anticancer Activity in HeLa Cells

Disease cancer caused by abnormal growth of tissue where there has been an error, fast and out of control. Judging from the fact of gender, more than 270,000 women die every year caused by cervical cancer. To inhibit the growth of cancer cells, a compound is needed that causes the cell cycle to stop so that the ability of cell proliferation decreases. Alkaloid compounds can inhibit proliferation through oxidative inhibition processes that can cause cancer. Mangrove plants have potential as anticancer, antimicrobial, and antioxidant. The content of chemical compounds found in mangroves are flavonoids, steroids, alkaloids, phenolites, saponins and tannins. These compounds show high antioxidant activity and are shown to have a real relationship with the properties of the material's bioactivity against cancer cells. One of the mangrove species is Rhizophora apiculata. The purpose of this study was to determine the IC50 value produced by Rhizophora apiculata mangrove leaf extract on HeLa cell viability and to see the effect of Rhizophora apiculata mangrove leaf extract dosage on HeLa cell viability. The method used in this research is the experimental method. The research parameters included yield, proximate test, phytochemical test, toxicity test, total phenol test, cytotoxicity test and LC-MS test. The experimental design used was a simple and complex completely randomized design (CRD) with the Tukey test.The results of this study showed that the highest yield was in the ethanol extract of 5.91%, while the n-hexane and ethyl acetate extracts respectively had yields of 1.18% and 1.31%. The results of the proximate test on the water content of leaves and powder were 64.53% and 13.86%, respectively, the results of the ash content in the leaves and powder of Rhizophora apiculata were 3.94% and 8.41%, respectively. while the water content in the extract obtained the highest yield in the ethanol extract of 21.42%, while the n-hexane extract and ethyl acetate extract were 11.08% and 15.42%, respectively. For phytochemical results, it was found that n-hexane extract only contained alkaloids, flavonoids and steroids. Ethyl acetate extract contains steroid compounds. Meanwhile, the ethanol extract contains the most bioactive compounds, namely saponins, flavonoids, tannins and triterpenoids. The toxicity test using the Brine Shrimp Lethality Test (BSLT) method resulted in the lowest IC50 of ethanol extract at 49.45 ppm while the n-hexane and ethyl acetate extracts were 251.63 ppm and 920.45 ppm respectively. In the total phenol test, the n-hexane extract was 66.79 mg GAE / 100 gr, 222.97 mg GAE / 100 gr ethyl acetate extract and 929.04 mg GAE / 100 gr ethanol extract. HeLa cell cytotoxicity testing using the MTT method (3- (4,5-dimethiltiazol-2-yl) -2,5-dipheniltetra zolium bromide) assay resulted in the highest cell viability value at a dose of 125 ppm of 46.97%. As for the doses of 250 ppm, 500 ppm 1000 ppm, and 2000 ppm resulted in a percentage of viability of 42.95% 37.70% 35.82% and 32.12%, respectively. The IC50 value of Rhizophora apiculata leaf extract was 64.42 ppm. This value indicates that the Rhizophora apiculata extract is toxic to HeLa cells.

The Effect Of Mangrove Leaf Extract Dosage Sonneratia Alba On Hela Cell Viability

Cervical cancer is caused due to infection from the Human Papilloma Virus (HPV) which attacks the sexually active female reproductive organs. Treatment is carried out alternatively using natural ingredients such as mangroves. Sonneratia alba is a type of mangrove plant that has been used in alternative medicine because of its potential as an anticancer. This study aims to determine the effect of Sonneratia alba mangrove extract on heLa cell viability. The Sonneratia alba leaf powder was extracted using stratified maceration. The solvents used include n-hexane, ethyl acetate, and ethanol. The results showed that the LC50 value was 3.59 ppm, this means that the ethyl acetate extract has toxic properties. Phytochemical test results of Sonneratia alba leaf extract contain alkaloid compounds, steroids / triterpenoids, and tannins. The results of the test yield extract were 12.60%, extract water content was 21.24%, and total phenol was 504.08 mg / g GAE Test The results of the LC-MS test resulted in the suspicion of compounds including diosmetin, caffeine, and turmeron. The ethyl acetate extract of Sonneratia alba leaves was cytotoxic against heLa cell viability with the resulting IC50 value of 478.630 µg / mL

The Effect Of Dosage Of Mangrove Leaf Extract Avicennia Marina On The Viability Of Hela Cells

Cervical cancer is caused by infection with the Human Papilloma Virus (HPV) which attacks the reproductive organs of sexually active women. Treatment is done alternatively using natural materials such as mangrove plants. Avicennia marina is a type of mangrove plant that has been used in alternative medicine because of its potential as an anticancer. This study aimed to determine the effect of Avicennia marina mangrove leaf extract on the viability of HeLa cells. Avicennia marina mangrove leaf powder was extracted using graded maceration. The solvents used include n-hexane, ethyl acetate, and ethanol. The results showed that the LC50 value was 98.55 ppm, it means that the ethanol extract has toxic properties. Phytochemical test results of Avicennia marina mangrove leaf extract contain saponins, steroids/triterpenoids, flavonoids and tannins. The test results showed that the extract yield was 14.40%, the water content of the extract was 16.57%, and the total phenol was 1915.92 mg/g GAE. The results of the LC- MS test resulted in suspected compounds including Caffeine and Diosmetin. The ethanol extract of Avicennia marina mangrove leaves was cytotoxic to heLa cell viability with the resulting IC50 value of 115.345 g/mL.

Effect Of Alpha-Tocopherol On Spermatozoa Death Of Apoptosis and Necrosis in Rats (Rattus Norvegicus) Exposed 2,3,7,8- Tetrachlorodibenzo-P-Dioxin

The aims of this research was to investigate effect of alpha-tocopherol on spermatozoa death in form of apoptosis and necrosis in rats (Rattus norvegicus) exposed 2,3,7,8- tetrachlorodibenzo-p-dioxin. Male rats were administered TCDD and alpha-tocopherol in experimental groups. Five experimental groups of a combination of TCDD and alpha- tocopherol were designed as follows; 0.5 ml of corn oil (control negative group, K-), 700 ng/kg/d of TCDD and 0.5 ml of corn oil (treatment control), 700 ng/kg/d of TCDD and 77 ng/kg/d of alpha-tocopherol (Group P1), 700 ng/kg/d of TCDD and 140 mg/kg/d of alpha-tocopherol (Group P2), 700 ng/kg/d of TCDD and 259 mg/kg/d of alpha- tocopherol (Group P3) respectively. Alpha-tocopherol and TCDD were given by oral gavage for 20 days. The result indicated that TCDD increased spermatozoa death in form of apoptosis and also necrosis. Alpha-tocopherol at 259 mg/kg/d most effective to decreased spermatozoa death number. The conclusion indicated that alpha-tocopherol at 259 mg/kg/d effective to decreased the spermatozoa death in form of apoptosis and necrosis in rats (Rattus norvegicus) exposed 2,3,7,8-tetrachlorodibenzo-p-dioxin.

Effect Of Mangrove Leaf Extract Dosage Rhizophora Mucronata Lmk. On The Viability Of Hela Cells

This study aims to determine the IC50 value of the mangrove leaf extract type Rhizophora mucronata Lmk. against the viability of Hela cells. The samples extracted with three types of solvents were previously conducted preliminary research on phytochemical compounds and their toxicity values with the Meted Brine Shrimp Letality Test (BSLT). In the toxicity test, the highest level of toxicity was obtained in the ethanol extract with a value of 166.72± 7.72 ppm, then the sample was continued for the cytotoxicity test using the MTT method. The dosage variants used in this study were 62.5 ppm; 125 ppm; 250 ppm; 500 ppm; and 1000 ppm. The variation in dosage shows an effect on the viability of Hela cells, namely a decrease in the percentage of living cells along with the addition of the ethanol extract dose of Rhizophora mucronata Lmk leaves. which is given. And the IC50 value obtained from this study was 63.67 µg / mL with the toxic category

Genetic Stability The Protein Encoding Envelope (E) Genes Dengue Virus Serotype-4 Passaged in Vero Cell As A Candidate Chimera Vaccine Material

This study aims to analyze genetic stability of the gene encoding the envelope protein (E) dengue virus serotype-4 passaged in vero cells, Denv-4 passaged in vero cells serially then continued with RNA extraction at passage 0, 10, 20, 30, 40 ,50 , and 60, and then continued with two step PCR and amplification, and sequencing then analyze the nucleotide stability with BLAST and MEGA 5 software. The result shows that there are many variable site in nucleotide and amino acid with high mutation rate 57.4% for nucleotide and 71.9% for amino acid ,while the similarity between passages are high ranging from 91% - 98%. The conclusion for this study is Denv-4 after analyzed shows that the gene encoding protein E has many variable site but high in similarity.

The Effect of Bone Marrow Transplantation on Oocyte-Granulosa Cell Interaction and Follicular Development of Cisplatin-Induced Ovarian Failure in Rat

Introduction: Chemotherapy has cytotoxic effect that induces follicular damage and abnormal folliculogenesis leads to ovarian failure. Two crucial growth factors in abnormal folliculogenesis, Growth Differentiation Factor-9 (GDF-9) and Kit-Ligand, will be disrupted and affect follicular development. In this study, we evaluate whether bone marrow transplantation (BMT) has a role on oocyte-granulosa cell interaction by analyzing GDF-9 and Kit-Ligand expressions and also follicular development by analyzing primordial, primary, secondary and graafian follicles of cisplatin-induced ovarian failure in rat. Material and method: Forty eight rats were divided into three groups: control, cisplatin and cisplatin + BMT. Ovarian failure was induced by administration of intraperitoneal cisplatin 5 mg/kg body weight for 1 week. BMT 2 x107 cells were injected through rat tail vein after cisplatin administration. Bone marrow was isolated from rat femur and characterized by CD44(+), CD45(-), CD105(+). Immunohistochemistry examinations for ovarian GDF-9, Kit-Ligand and follicle development evaluation were performed after 2 weeks of BMT injection. Results: The expressions of Kit-ligand among three groups by ANOVA were significant different (p=0.00), whereas by Post Hoc: cisplatin group lower than control group (p=0.00); cisplatin + BMT group higher than cisplatin group (p=0.00); and no significant different between control group and cisplatin + BMT group (p=0.955). The expressions of GDF-9 by Kruskal Wallis showed significant different (p=0.00) among three groups whereas cisplatin + BMT group higher than cisplatin group and control group. In cisplatin + BMT group the number of primordial, primary, secondary and graafian follicles were higher than those in cisplatin group; but were lower than those in control group (p=0.000). Positive Paul Kart Horan (PKH) labeling was seen in cisplatin + BMT group only. Conclusion: In cisplatin-induced ovarian failure in rat, bone marrow transplantation may improve oocytegranulosa cell interaction and follicular development.

The Therapy of Burn Wound Healing in Rat (Wistar) by Using the Combination of Peripheral Blood Mononuclear Cells (PBMCs) and Bone Marroe BM-Derived Mesenchymal Stem Cell

The therapy to heal the bum wound is still imperfect, therefore it is important to conduct specific research concerning this topic that benefits to society. Using both Bone Marrow-stem cell (BM-MSc) and peripheral blood mononuclear cells (PBMCs) from allogenic donors as part of the therapy to heal the bum wound seems to give positive prospect for the future treatment. In this experiment, PBMC and rat BM- derived from mesenchymal stem cell were used as the therapy model. lmmunocytochemistry was used as the method to characterize the phenotype of MSc, It was also used to express the collagen type I and the Indirect ELISA in analyzing the TGF·P 1 secretion. The rats with bum wound were divided into 2 kinds of group; the first group of rats was selected to control the use of PBS; while second group of rats was used as the treatment object that was medicated by the applying of the combination of both BM-MSC and PBMC. Stem-cells subcutaneously administered dose applied to each rat was around of 2 x 106 cells. The result showed that the levels of TGF-β1 secretion in day 3rd and day 7th on the rats which were treated by using the combination of BM-MSC and PBMC were higher compared to the rats from the control group. The experiment that concerns on the thickness of the collagen showed that combination between BM-MSC and PBMC stem cell make it possible in increasing the thickness of collagen 1, besides; it also showed significant differences (p=0.000). This research proved that the combination of BM-MSC and PBMC stem cell served can accelerate the healing process for the bum wound on rats through Increasing of TGF-P 1 secretion and collagen type 1 expression, It means that PBMCs can be applied as good as chemoattractant.

Characterization of CD4+ Lymphocyte From Bone Marrow Stem Cell Using Indirect Immunofluoresence For HIV & AIDS Treatment

Acquired immune deficiency syndrome (AIDS) is caused by Human Immunodeficiency Virus (HN). At the beginning of infection, gpl20 virus interacts with CD4 receptor at the surface of the target cell. The interaction between gpI20 and CD4 leads to the occurrence of the binding of specific chemokine receptor CXCR4 and CCR5, which are also present on the membrane of the target cell. Therefore, CCR5 arid CXCR4 also determine the fate of the target cell. It is the performance of CCR5 and CXCR4, guided by controlling gene that determines susceptibility or resistance to HN infection. Coding gene CCR5 may mutate to become protective or resistant against HTV infection. In homozygote individuals, it tends to be resistant against infection while in heterozygote individuals it tends to be susceptible to HN infection. Objective: To characterize TCD4 lymphocyte in the next that is resistant against HN infection by using gene therapy deletion 32 CCR5 to use for HTV & AIDS treatment. Method: Sample collection, mononucleated cell collection, lymphocyte culture, CD4 identification, CCR5 variance analysis, co-cultivation with PBMC HN and comparison to control. Result: This study was performed in several steps, such as mononucleated cell isolation, followed with cell culture, lymphocyte purification, lymphocyte and CD4 expression identification. Conclusion: Lymphocyte T CD4 had been mature after seven passages, once passage is about 5 days so for maturity lymphocyte T CD4 need 35 days and that cell as be candidate to resistant against HN infection by using gene therapy deletion 32 CCR5 to use for HN & AIDS treatment.

The Role of adaptive MSCs as an Attempt Regeneration of Spermatogenesis Process Using by Hypoxia Precondition In Vitro

The aim of this research was to obtain to get adaptive MSCs by an treatment in hypoxia precondition in viro culture. Ini this research, hypoxia precondition was to be given 3% O2 concentration and compared to those of normoxia culture in O2 > 20%. Results showed that under 3% O2 concentration, CD105+ and CD45- by flowcytometri, immunocytokimia and immunofluorescence didn’t experience of change (undifferentiated). Meanwhile under > 20% O2 concentration, cells have experienced of change (not undifferentiated again), that was indicated with CD105+ become decrease and CD45- increase by flowcytometri, immunocytokimia and immunofluorescence. Conclusion, in this research showed that hypoxia precondition with 3% O2 concentration very support MSCs to constantly adaptive before transplantated for disturbance of spermatogenesis process, because did’nt experience become progenitor cells was not expectation (still undifferentiated).