Triana Hertiani

Antimicrobial effects of Indonesian Medicinal Plants Extracts on Planktonic and Biofilm Growth of Pseudomonas aeruginosa and Staphylococcus aureus

Objective: The increasing rates of antibiotic-resistant microbial infections requires continuous development of new antimicrobial agents. Moreover, microbial biofilms exhibit elevated resistance to most antimicrobial drugs and the host defense systems, which often results in persistent and difficult-to-treat infections. The discovery of anti-infective agents which are active against both planktonic and biofilm microbial are consequently required to deal with these biofilm-mediated infections. The aim of this study is to evaluate the activity of Indonesian medicinal plants extracts on planktonic and biofilm growth of Pseudomonas aeruginosa PAO1 and Staphylococcus aureus Cowan I. Methods: Fifty four (54) ethanol extracts were obtained from a variety of known Indonesian medicinal plants. The growth inhibitory concentration (MIC), effects on biofilm formation and biofilm breakdown, and biofilm architecture in the absence and presence of the extracts by confocal laser-scanning microscopy along with LIVE/DEAD staining was performed. Results: Plantextracts showed an inhibitory effect on planktonic growth of these bacteria and also on their biofilm formation. At a concentration as low as 0.12 mg/ml, biofilm formation of P. aeruginosa PAO1 and S. aureus Cowan I is inhibited by 5 plant ethanol extracts: Kaempferia rotunda L., Caesalpinia sappan L., Cinnamomum burmanii Nees ex Bl., C. sintoc and Nymphaea nouchali Burm. f. Limited bacteriostatic activity was evident. Conclusion: The results clearly indicate the extracts obtained are interesting sources of putative antibiofilm agents. This research can contribute to the development of new strategies to prevent and treat biofilm infections.

Potency of Massoia Bark in Combating Immunosuppressed-related Infection.

Background: As part of our search for new potential natural resources to eradicate infection, we have revealed the prominent potency of massoia bark (Massoia aromatica Becc, Lauraceae) in combating immunosuppressed-related infection. Materials and Methods: The extract was prepared by macerating the pulverized dried bark in ethanol 95%, followed by solvent evaporation. The oil was extracted from the dried bark by steam-hydrodistillation of which preparative thin-layer chromatography was performed on the oil to isolate the active constituent, C-10 massoia lactone (ML). Anti-biofilm assay against Candida albicans was conducted on polystyrene 96 wells microtiter plates, followed by a confocal laser scanning microscope observation to get three-dimensional profiles of the affected biofilms. Effects on the hyphae development inoculated on RPMI-1640 agar plates were observed for 7 days. Influences of samples on mice macrophage phagocytosis were examined by an in vitro technique. Samples concentration tested were in the range of 2.0–0.0625 mg/mL and done in triplicate. Results: Massoia bark extracts (oil and solid phase) and ML exhibited promising activities as anti-biofilm against C. albicans at IC50 0.074% v/v, 271 μg/mL and 0.026 μg/mL, respectively. The ML did not inhibit the hyphae development at the concentration tested; however, the extracts showed inhibition at 62.5 μg/mL. Macrophage phagocytosis stimulation was correlated to the ML content. Conclusion: Massoia bark is potential to be developed as anti-infective in immunosuppressed condition of which the C10 ML (C10H16O2) plays a major role in exerting activity.

Ant Plant (Myrmecodia tuberosa) Hypocotyl Extract Modulates TCD4+ and TCD8+ Cell Profile of Doxorubicin-Induced Immune-Suppressed Sprague Dawley Rats In Vivo

Myrmecodia tuberosa Jack (Rubiaceae) has been used as part of traditional Indonesian remedies for a wide range of therapeutic usages in West Papua. Our preliminary study revealed the significant potency of these plant extracts and fractions as an immunomodulator by an in vitro technique on Balb/c mice. This study explored the effect of M. tuberosa hypocotyl ethanol extract on the TCD4+ and TCD8+ cell profiles of doxorubicin (Dox)-induced immune-suppressed Sprague Dawley (SD) rats by an in vivo method. Dried powder of M. tuberosa hypocotyl was macerated in 95% ethanol. Following solvent evaporation in a vacuum, the ethanol extract (EE) was partitioned to yield an n-hexane fraction (FH) and residue (FNH). FNH was further partitioned to yield ethyl acetate (FEtOAc) and water fractions (FW). The extract and fractions in the concentrations 10, 20, 50, and 100 μg/mL were tested on macrophage cells by the latex bead method, while the proliferation of lymphocyte cells was evaluated by the MTT assay. The total phenolic and flavonoid contents of those fractions were evaluated. The active fraction was administrated orally on Dox-induced SD rats for 28 days by an in vivo method to observe the TCD4+ and TCD8+ cell profiles. The in vivo assay showed that the FNH could maintain the number of TCD4+ cells, but not the number of TCD8+ cells. The ED50 observed was 24.24 mg/kg BW. Steroid/terpenoid compounds were detected in this fraction along with the phenolics and flavonoids. The FNH contained 3.548 ± 0.058% GAE of total phenolics and 0.656 ± 0.026% QE of total flavonoids. M. tuberosa hypocotyl extract is a potent immunomodulatory agent and may act as co-chemotherapy in Dox use.

In vitro Evaluation of Massoia Bark Essential Oil and C-10 Massoialactone Potency as Immunomodulator

Abstract Massoia aromatica Becc (Lauraceae) bark essential oil has been widely used as part of traditional healing remedy in Indonesia. The major constituent, C-10 Massoia lactone has been reported previously as the antimicrobial principle. However, its effect on the immune response has not been reported so far. This research was aimed to find out the immunomodulatory effects of the Massoia essential oil and the C-10 Massoia lactone against Candida albicans as well as to explore the potential toxicities against Vero and primary culture of fibroblast cells. Material and methods: The essential oil was obtained by a steam-hydrodistillation method. The C-10 Massoia lactone was isolated from the essential oil by a preparative chromatography. Cytotoxicity assays were done on Vero and primary culture of fibroblast cells by using the MTT assay. The immunological assay were performed in vitro on lymphocytes and macrophages taken from Sprague-Dawley mice. The concentration range of samples were 30 - 2.5 μg/mL and done in triplicate. ANOVA followed by the Tukey-HSD (honestly significance diffirence) was used for the statistical analysis. All sample showed significant activation of the macrophage phagocytosis both on latex and Candida albicans in a concentration dependant manner. IC50s of the C-10 Massoia lactone and the essential oil on primary culture of fibroblast were 11.29 and 26.07 μg/ml, respectively, while on Vero cells were found at 28.35 and 37.34 μg/ml, respectively. All samples exhibited no effect on lymphocyte cell proliferation.

Effect of Massoia (Massoia aromatica Becc.) Bark on the Phagocytic Activity of Wistar Rat Macrophages

The essential oil of Massoia (Massoia aromatica Becc., Lauraceae) bark is a potential immunomodulator in vitro. This study evaluated the potential immunomodulatory effects of Massoia bark infusion on the nonspecific immune response (phagocytosis) of Wistar rats. For the in vitro assay, macrophages were treated with the freeze-dried infusion at the concentrations of 2.5, 5, 10, 20, or 40 µg/mL media. For the in vivo assay, two-month-old male Wistar rats were divided into five groups. The baseline group received distilled water at the dose of 1 mL/100 g body weight (BW), with the herbal product containing Phyllanthus niruri extract that was administered as the positive control at the dose of 0.54 mL/rat. The treatment groups received the infusion at a dose of 100, 300, or 500 mg/100 g BW. Treatments were given orally every day for 14 days. The ability of macrophage cells to phagocyte latex was determined as phagocytic index (PI), and it was observed under microscopy with 300 macrophages. The in vitro study revealed that the phagocytic activity of the infusion-treated macrophages significantly increased in comparison with that of the control macrophages in a concentration-dependent manner. Among all of the treatment concentrations, the concentration of 40 µg/mL provided the highest activity with a PI value of 70.51 ± 1.11%. The results of the in vivo assay confirmed those of the in vitro assay. The results of the present study indicate that Massoia bark can increase the phagocytic activity of rat macrophage cells.

HUBUNGAN PENGETAHUAN, PERSEPSI KUALITAS, DAN NIAT APOTEKER UNTUK MEREKOMENDASIKAN FITOFARMAKA

Apoteker yang mempunyai pengetahuan yang baik dan persepsi yang positif terhadap kualitas fitofarmaka diharapkan akan merekomendasikan produk fitofarmaka kekonsumen di apotek. Penelitian bertujuan untuk mengetahui gambaran pengetahuan dan persepsi apoteker terhadap kualitas fitofarmaka, serta pengaruhnya terhadap niat merekomendasikan produk fitofarmaka. Penelitian merupakan penelitian korelasional dengan instrumen berupa kuesioner. Responden adalah apoteker yang bekerja di apotek yang menjual fitofarmaka di Yogyakarta. Kuesioner terdiri dari 4 bagian yaitu karakteristik responden, pengetahuan fitofarmaka, persepsi kualitas fitofarmaka, dan niat merekomendasikan produk fitofarmaka. Persepsi kualitas dan niat merekomendasikan diukur dengan skala Likert. Teknik pengambilan sampel dilakukan secara acak sederhana. Data dianalisis dengan analisis deskriptif dan regresi linier sederhana. Hasil penelitian menunjukkan bahwa pengetahuan apoteker di apotek kota Yogyakarta secara rata-rata adalah cukup (nilai mean 6,67). Persepsi apoteker tentang kualitas fitofarmaka adalah baik (positif) berturut-turut dari nilai dimensi tertinggi adalah keamanan, efikasi, ketersediaan, akseptabilitas, merek, dan harga. Pengetahuan apoteker tentang fitofarmaka memberikan pengaruh positif dan signifikan terhadap persepsi kualitas fitofarmaka. Pengetahuan dan persepsi apoteker tentang kualitas fitofarmaka masing–masing memberikan pengaruh positif dan signifikan terhadap niat merekomendasikan produk fitofarmaka. Kata kunci : fitofarmaka, pengetahuan, persepsi kualitas

TOXICITY TEST OF MAKUTADEWA (Phaleria macrocarpa (Scheff.)Boerl.) BARK AGAINST Artemia salina Leach AND THIN LAYER CHROMATOGRAPHY PROFILE OF THE ACTIVE FRACTION

Makutadewa (Phaleria macrocarpa (Scheff) Boerl.) is one of the herbal medicines used against cancer, however, the scientific basis of the activity still void. Daphne mezereum, the same family (Thymelaeaceae) to makutadewa has been proved to contain cytotoxic substances in its bark. Therefore, the aims of this research were to investigate the toxic effect of makutadewa bark against Artemia salina Leach. as the primary step to identify an anticancer activity and find out substances responsible to this activity. The material powder was extracted using Soxhlet apparatus with chloroform, followed by methanol and finally by distilled water. The toxicity of chloroform, methanolic and aqueous extracts were assayed against A.salina Leach. The chloroform extract (most active) with the LC50 of 29.6  0.14 g/ml was fractionated by vacuum liquid chromatography using wash benzene 100 % (a); wash benzene-ethyl acetate =20:1 (b); 15:1 (c); 10:1 (d); 5:1 (e); 2:1 (f) and chloroform-methanol 1:1 (g) as mobile phases. The active fractions against A.salina were E {combination of (e) and (f)} with LC50 of 106,9 g/ml and F (g) with the LC50 of 131,53 g/ml, hence less toxic than the original extract. The thin layer chromatogram profiles showed that E and F fractions contained terpenoid and alkaloid substances. Key words : toxicity test , (Phaleria macrocarpa (Scheff)Boerl.), bark

TOXICITY OF ETHANOLIC EXTRACT OF FRUIT, SEED AND LEAVES OF MAKUTADEWA (Phaleria macrocarpa (Scheff.) Boerl.) TO Artemia salina Leach AND THIN LAYER CHROMATOGRAM PROFILE OF ACTIVE EXTRACT

A research has been done to determine the toxicity of ethanolic extract of fruit, seed and leaves of makutadewa (Phaleria macrocarpa (Scheff) Boerl.) using Brine Shrimp Lethality Test method. Extract were made by maceration of the materials for 24 hours. The bioassay has been done using 48 hour old Artemia saline Leach. The effect of ethanolic extract was identified by determining and analysing the shrimp death percentage and LC50 of each extract using probit analysis. The result showed that these ethanolic extract were toxic to the shrimp. The ethanolic extract of seed was the most active with the LC50 of 1.60 x 102 g/ml, whereas that of the fruit has the LC50 of 30.42 g/ml. The thin layer chromatogram profile of the active extract suggested that the extract contains alkaloids. Key words : toxicity, makutadewa, brine shrimp lethality test