Produksi senyawa bioaktif antimikroba pada Dumortiera hirsuta dengan bioteknologi kultur jaringan : Identifikasi senyawa bioaktif antimikroba pada lumut hati (Dumortiera hirsuta)

Lurnut hati Dumortiera hirsuta memiliki potensi sebagai antimikroba. Tujuan penelitian ini adalah mengetahui aktivitas antimikroba ekstrak kalus dan mekanisme penghambatan senyawa antimikroba ekstrak kalus terhadap mikroba patogen Staphylocossus aureu.{ ATCC 25923, Escherichia coli ATCC 25922, dan Candida albicans ATCC 10231. Aktivitas antimikroba diuj i dengan metode difusi cakram dan pengenceran dalam tabung. Data diameter zona hambat dan nilai MIC dianalisis secara deskriptif Ekstrak kalus dianalisis dengan GCMS. Berdasarkan hasil peneltian ekstrak kloroform menunjukkan aktivitas antimikroba yang tinggi terhadap Candida albicans dibandingkan dengan Staphylococcus aureus dan Escherichia coli. Mekanisme penghambatan senyawa antimikroba ekstrak kalus terhadap mikroba patogen adalah dengan merusak biomembran mikroba

ISOLASI FRAKSI SITOTOKSIK IN VITRO TERHADAP SEL KANKER LEHER RAHIM (HeLa) DARI EKSTRAK KLOROFORM BUAH MERAH (Pandanus conoideus Lamk.) VARIETAS MALER

Recently buah merah has been extensively used to treat cancer. The scientific studies of anticancer activity of buah merah on the growth of cancer are relatively very few. It was found that the best extract that had the highest toxicity on breast cancer and cervic cancer cells was chloroform extract of Maler. But the compound of those extract has not be identified and isolated. Therefore the objective of these studies were to identify and isolate the in-vitro cytotoxic fraction on cervic cancer cells from chloroform extract of buah merah (Pandanus conoideus Lamk.) Maler variety. Buah merah local variety Maler and cancer cell line HeLa were used in this study. Buah merah was extracted using chloroform. The extract was fractionated using vacuum coloumn chromatography followed by TLC. Cytotoxicity was performed using MTT assay. The IC50 was determined using linear regression analysisis.The toxic compound of the most toxic fraction was separated using preparative TLC. The products of preparative TLC were the lower position, middle position and upper position were tested using a MTT assay. Profile of the highest cytotoxic fraction was monitored by TLC using n-hexan (4 ml) : aceton (4 ml) : glacial acetic acid (6 drops) as a solvent system. This study showed that the low position of preparative TLC profile had the lowest IC50 (109.99 µg/mL). Chloroform extract of buah merah Maler variety 75.20 gram could produce 0.166 gram cytotoxic fraction. Chromatogram TLC of those cytotoxic fraction showed that the profil was similar to toxic compound on breast cancer T47D. It could be concluded that the cytotoxic fraction in chloroform extract of buah merah Maler variety could be separated using preparative TLC. The ratio of extract and cytotoxic fraction on cervic cancer was 453 : 1

IDENTIFIKASI GOLONGAN SENYAWA ANTIMIKROBA PADA LUMUT HATI Dumortiera hirsuta Nees. DAN PERBANYAKANNYA DENGAN KULTUR JARINGAN

The resistance of infectious microbes is increasing. One alternative of reducing this resistance is by exploring plant-based natural anti-microbes on bryophytes. Based on the previous research, acetone and methanol-water extract of Dumortiera hirsuta in have shown inhibitory activities as antibacterial and antifungal. However its active compound that have the highest inhibitory activities as antibacterial and antifungal have not been elucidated. The problem of using the liverwort as a source of antimicrobial compound was its low availability. Therefore, the objective of this research was to identify the class of bioactive antimicrobial compound in liverwort and to produce the antimicrobial compound by plant-tissue culture. In this research, Dumortiera hirsuta that was taken from Cangar forest, Batu as source of antimicrobial compound. Staphylococcus aureus ATCC 25923,Escherichia coli ATCC 25922, and Candida albicans ATCC 10231 collections of laboratory of Biology Department, Faculty of Science and Technology, Airlangga University were used as test microbes. Dumortiera hirsuta was extracted using three solvents chloroform, methanol, water. Antimicrobial activity of extracts were tested by using disc diffusion and tube dillution methods. The best extract was fractioned by Vaccum Liquid Chromatography and bioactive compounds are monitored by using Thin Layer Chromatography (TLC). The most potential activity of the fractions was determined by the same methods. The best fractions with highest inhibitory activities will be analyzed by Preparative-TLC to detect the bioactive compound class. The explants of sporophyte and gametophyte with various size (0,

EFEK INFUSA BIJI KEMRUNGGI (Caesalpinia crista L.) TERHADAP KADAR GLUKOSA DARAH DAN SGPT, SERTA STRUKTUR HISTOLOGIS HATI DAN GINJAL TIKUS PUTIH (Rattus norvegicus Berkenhout, 1769) HIPERGLIKEMIK

The objective of this research was to know the effect of Kemrunggi (Caesalpinia crista L.) seeds infusion on blood glucose, SGPT level, histological of liver and kidney of hyperglycaemic rats (Rattus norvegicus Berkenhout, 1769). Kemrunggi (C. crista L.) seeds were collected from West Papua and rats (R. norvegicus Berkenhout, 1769) male, Wistar. Rats were divided into 7 groups of 5, 4 groups as control (independent, burden, solvent, and drug) and 3 groups used as a treatment groups that were given 10%, 20% and 30% Kemrunggi seeds infusion. The hyperglycaemic rat were made by administering D-glucose monohydrate 6,75 g/kg body weight. The seeds was infundation and was treats to hyperglycaemic rats during seven days. The result showed that on the seventh days, blood glucose level was lowest on independent group (AUC0-300 32421,32). There was no significantly different among infusion treatments, eventhough the 20% infusion group showed the highest percentage of lowering blood glucose value (55,282%). No significantly different among treatments also showed on the lowering SGPT level, at 10%, 20%, and 30%, were 26,13%, 10,02%, and 17,55% respectively. Kemrunggi (C. crista L.) seeds infusion were caused degeneration on liver histological (cloudy sweeling, hydrophic), steatosis, and karyolisis, but showed no significantly different with control group. It also were caused degeneration on kidney histological (cloudy sweeling, hydrophic), plasmolysis, and karyolysis. It could be concluded that infusion of Kemrunggi seeds at 20% lowering blood glucose effectively and also lowering SGPT level at 10-30% concentration. Kemrunggi seeds infusion did not have any effect on liver histological structure, but not on kidney

IDENTIFIKASI GOLONGAN SENYAWA PENURUN KADAR GLUKOSA DARAH TIKUS PUTIH (Rattus norvegicus Berkenhout, 1769) HIPERGLIKEMIA PADA DAUN SUKUN (Artocarpus altilis (Park.) Fosberg.)

Sukun (Artocarpus altilis (Park.) Fosberg.) leaf has been extensively used to treat various health problems including diabetes mellitus. . The scientific studies of hypoglycemic activity of breadfruit leaves are relatively very few. Therefore the objective of this research was to identify the bioactive compound that have activity to lower blood glucose. Forty hyperglycemia rats, male Wistar 2 month old were used in this study. Hyperglycemia rats were made by injecting glucose monohydrate 1,35 g/kg body weight. For extract efectivity test, 20 rats were divided into 4 groups of 5. One group as a control, the next two groups were given extract of chloroform and methanol respectively, and the last group were given boiled water extract of sukun leaves. The best extract was fractionated by partition using ethyl acetate. Another 20 rats were used to analyze the activity of the fraction. They were again divided into 4 groups of 5. One group as a control, another one group were given glibenclamide as a reference, and another two groups were given 2 kinds of fraction (water fraction and ethyl acetate fraction). The dose that used for each extract, boiled water extract of sukun leaves, and fraction of the best extract were at 150 mg/kg body weight. Blood glucose was determined at minute 0, 60, 120, 180, 240, and 300. Ethyl acetate fraction of boiled water extract was analyzed using Thin Layer Chromatography followed by application of various detection reagents for identification of compounds.. ANOVA was used to analyze the difference among samples followed by LSD 5% to locate the difference. The results showed that boiled water extract of sukun leaves has the highest activity, followed by chloroform extract. Both ethyl acetate and water fraction of boiled water extract of sukun leaves could lower the blood glucose, but were not as good as glibenclamide. The bioactive compound groups that have hypoglycemic activity were classified as phenolic, flavonoid, alkaloid and terpenoid. It could be concluded that traditional used of sukun leaf boiled water to treat diabetes mellitus has been already proper way

EFEK EKSTRAK DAUN TEH (Camellia sinensis (L.) Kuntze) TERHADAP GAMBARAN HEMATOLOGI, KADAR NO, AKTIVITAS GPx PADA INDIVIDU DENGAN POLIMORFISME GENA NOS3 DAN δ-ALAD YANG TERPAPAR PLUMBUM

Lead have used as ceramics, fireproof materials and additive agent of gasoline. Lead is toxic to human especially for lead-exposed individual. Green tea has been extensively used to treat many kind of disease because its high contain antioxidant called catechin. The objective of these studies were to know the effect of green tea as a chelating agent of Pb. This research was conducted in two stages. The first stage was to know the best green tea formulation for treatment using Wistar rat Fourty two male Wistar rats were used in this studies. Rats were devided into 7 groups of 6. First group as a negative control. The rest were the treatment groups that were given Pb acetate orally for 66 day. On the 35 th day of studies group II, III, IV and V were given leaf green tea extract at 0 g/kg/bw/day (positive control), 0.75 g/kg/bw/day, 1.5 g/kg/bw/day, 3 g/kg/bw/days respectively. The group VI was given green tea infusion 5 g/kg/bodyweight/days and the group VII was given dimercaprol 3 g/kg/bw/days, as a standard clinical medicine. Parameter recorded were hematological profile that measured by hematology analyzer and blood lead, malondialdehyde, glutathione peroxidase activity, nitric oxide that measured by commercial kits. The different among treatments were analyzed using ANOVA, followed by Duncan to locate the different. The supporting parameters histopathological liver and kidney of rat, was observed on 66 th day, urine Pb levels and body weight of rats was observed on 0 and 66 th day. These results showed that treatment of all doses of green tea extract, green tea infusion and dimercaprol could decrease Pb and MDA level and increase NO level and GPx activity. The groups of rats that treated with green tea solution dose 5 g/kg/bw/day, gave a better histological figure compare with all of rats group. Based on these results, green tea infusion dose 5 g/kg/bw/day is the best treatment. The second stage of studies using a survey method with a cohort design These subject were 30 autorepair workers as cases subject and 30 control subject come from non polluted area (Pamijen village). The genotype individu polymorphisms NOS3 and ALAD gene were analyzed by Polymerase Chain Reaction (PCR)- Restriction Fragment Length Polymorphisms (RFLP) with MBoI restricted enzyme for NOS3, MspI restricted enzyme for ALAD gene. Parameter recorded were hematologic profile, blood lead level, malondialdehyde level, glutathione peroxidase activity and nitric oxide level. Suppporting parameters include systolic and dyastolic blood preassure. Data were analyzed by pair t-test

IDENTIFIKASI GOLONGAN SENYAWA ANTIHIPERGLIKEMIK Caesalpinia globulorum Bakh. f. & Van Royen PADA TIKUS PUTIH (Rattus norvegicus L.) HIPERGLIKEMIA

Caesalpinia globulorum Bakh. f. & Van Royen is an original plant in Buton island, South east Sulawesi. The benefits of this plant are known rarely by local people, whereas this plant has potential as a herbal medicine. In India Caesalpinia bonducella have used as antihyperglycaemia. Therefore the objective of this study was to identify antihyperglycaemic compound of Caesalpinia globulorum Bakh. f. & Van Royen on hyperglycaemia rat. The Caesalpinia globulorum Bakh. f. & Van Royen seeds were collected from Buton island. Hyperglycaemia rats (Rattus norvegicus L.) male, wistar were used in this study. The hyperglycaemia rat were made by injecting glucose monohydrate 1,35 mg/kg BB. The seed was extracted using ethanol, to detect the effectiveness of ethanolic extract, 200 mg/kg BB extract were given to hyperglycaemia rat using two methods. They were preventive method (exstract were given at initial treatments) and curative method (exstract were given at 45 th minutes). Ethanolic extract of Caesalpinia globulorum Bakh. f. & Van Royen seed was triturated by n-heksan. The yield of trituration were liquid fractions and solid fractions. The effectiveness of both fractions were detected using the same method as in the extract detection. Both of an extract and fractions were analyzed using thin layer chromatography followed by application of various detection reagents for identification of compounds. The results showed that the extract given to the preventive group lowered blood glucose level significantly compared with other groups. The extract (in preventive group) had the lowest AUC0-300 was 4085,5 and the highest percentage of lowering blood glucose (10,75%). The effectiveness of liquid fractions showed that at the preventive group the AUC0-300 was 4526,32 and the percentage of lowering blood glucose was -14,46 %. The AUC0-300 of liquid fraction in curative group was 4194,17 and the percentage of lowering blood glucose was -3,59 %. While the AUC0-300 of solid fraction (in preventive group) was 4585,72 and the percentage of lowering blood glucose was -16,35 % and the AUC0-300 in curative was 4472,87 and the percentage of lowering blood glucose was -13,05 %. The glibenklamid group had the lowest AUC0-300 was 3659,47 and the highest percentage of lowering blood glucose was 12,77%. The identification of antihyperglycaemic compound showed that both either extract or fraction was terpenoid. It could be concluded that the antihyperglycaemic compound on hyperglycaemia rat, in the ethanolic extract of the seed of Caesalpinia globulorum Bakh. f. & Van Royen was terpenoid which has a few conjugated double bonds

KAJIAN PENGARUH CHITOSAN TERHADAP PROFILLIPOPROTEIN, FUNGSI ENDOTELIAL AORTA DAN ARTERIA KORONARIA TIKUS PUTIH SPRAGUE DAWLEY DIINDUKSI LEMAK TINGGI

Atherosclerosis, the most common cause of death in developing country is a complex process involving the interplay of genetic and environmental factors and the involvement of multiple cell types. Injury to the vascular endothelium is thought to iniate the atherosclerotic lesion. The objective of this study was to analyze the influences of chitosan on lipoprotein profile, endothelial function of aorta, endothelial function of coronary artery, and C-Reactive Protein in rats by high fat induction. Twenty male Sprague Dawley rats, 1,5 months of age were used as experimental animals. All rats were adapted in 20 single cages for 7 days and given basal diet containing normal fat and water ad libitum. The rats were divided into 4 groups of 5. Group I was fed basal diet containing normal fat for 3 months. Group II, III and IV was fed high fat diet. Group II was used as a control (no treatment). Group III was fed chitosan 180 mg kg BW per day. Group IV was given chitosan using the same doses as group III, one month after feeding high fat diet. Chitosan was given orally in 2 ml aquadest. After 90 days, each rat was fast for 12-14 hours before blood collected. Blood was collected from the orbital vein for lipoprotein profile analysis. The rats were then sacrificed and the heart including aorta were collected for histopathological and immunohistochemical (expression of CRP) analysis. The difference between treatment was statistically analyze by Anava, and continued by LSDT to locate the difference. The result showed that high fat diet decreased HDL level, apoprotein A concentration, endothelial function of aorta and coronary artery, but increased total cholesterol level, LDL level and apoprotein B concentration. Chitosan increased HDL level and apoprotein A concentration, but decreased total cholesterol level, LDL level and apoprotein B concentration in rats by high fat induction. High fat diet increased coronary artery wall thickness and chitosan could be decreased coronary artery in rats. High fat diet + chitosan that given simultaneously could be increased coronary artery lumen diameter in rats. All those results were statistically significant (P<0,05). High fat diet did not significantly decreased coronary artery lumen diameter, as well as the rats that given chitosan after one month. Histopatologic analysis of aorta and coronary artery of rats that given normal diet showed that there was no atherosclerotic lesions, whereas aorta in rats fed high fat diet (P1) showed atheroma plaques. Structural illustration of aorta and coronary artery in rats fed high fat diet + chitosan (P2) did not show atherosclerotic lesions, whereas the group of rats that given chitosan after 1 month (P3), also did not show atherosclerotic lesions, in spite of several of aortas were atheroma plaques illustration. Immunohistochemistry analysis using antibody anti CRP of aortas showed that expression of CRP negative for all groups. CRP expression of coronary artery of rats that given normal diet (K) were negative, whereas rats that given high fat diet (P1) and no treatment were positive. CRP expression of coronary artery for rats that given high fat diet and given chitosan (P2) were negative, but rats that given chitosan after one month (P3) were positive. The overall results can be said that: 1. High fat diet decreased HDL level, apoprotein A concentration, endothelial function of aorta and coronary arter

BIOSISTEMATIKA PAMELO (Citrus maxima (Burm.) Merr.) DI INDONESIA BERDASARKAN KAJIAN MORFOLOGIS, FITOKIMIA, DAN MOLEKULAR

Pummelo (Citrus maxima (Burm.) Merr.) is native fruit of tropical South East Asia regions. There are a number of commercial pummelo cultivars intensively cultivated in various orchards in Indonesia. Based on the information found in many literature, pummelo is known as having high morphological diversity, especially in fruit characters. Nevertheless, so far there is no comprehensive publication documenting this diversity as well as the taxonomic treatment at infraspecific level. Systematics study reported here used both phenetic and phylogenetic approaches to disclose infraspecific variation, taxonomic affinitity, and clarification of taxonomic identity of pummelo at infraspecific level based on morphological, phytochemistry, and molecular analysis. The objectives of this study were to generate open classification system a infraspecific level for Indonesian pummel, and reveal morphological, phytochemical, and molecular markers as basis for the infraspecific classification. Materials used in this study were vegetative and reproductive organs of 107 individuals. Morphological data from leaves, flowers, fruits and seeds were obtained from observation and measurements either in the field or laboratory. Phenetic affinity based on morphology was conducted on 60 accessions using 21 characters. Meanwhile, phenetic affinity based on leaf essential oils was performed on 40 accessions using compounds� composition and concentration analyzed using gas chromatography � mass spectrometry. Morphological and essential oil data were subjected to cluster analysis and principal component analysis. The molecular approach was conducted using DNA sequence analysis of ITS1 on 70 accesions and (GA)n repeat motifs of microsatellites on 80 accessions. Phylogenetic relationships was examined using minimum evolution method. Result of cluster analysis on morphological data revealed two main clusters corresponding to the existence of two cultivar groups of pummelo. Six morphological characters from leaves and fruit were identified as the major distinguishing characters for these two clusters based on principal component analysis. Results of morphological analysis as used as a basis for constructing open classification in which 60 accessions of pummelo was divided into two cultivar groups, the �Gulung Cultivar Group� and �Pamelo Cultivar Group�. Combination of leaf and fruit characters was found to be morphological marker for the recognition of cultivar groups. Analysis on the GC-MS chromatograms of 40 pummelo accessions revealed infraspecific variation on leaf essential oil. The qualitative and quantitative infraspecific variation was used to differentiate accessions into three chemotypes. They were recignized as �nerol-loliolide-allo aromadendrene�, �alpha selinene�, and �alpha pinene-delta carene� chemotypes, respectively. Analysis on the ITS1 sequence showed a low infraspecific variation of 1.83%, and thus supported the role of ITS1 as molecular marker for differentiating species. Meanwhile, sequence analysis on microsatellite containing (GA)n dinucleotide repeat discloses high genotypic variation on pummelo. This high genotypic variation was reflected in phylogeny tree constructed by minimum evolution method which showed the existence of 9 clades