Isolation of active compounds from Streptomyces sennicomposti GMY01 and cytotoxic activity on breast cancer cells line.

The occurrence of resistance to anticancer and the emergence of serious side effects due to chemotherapy is one of the main problems in cancer treatment, including breast cancer. The need for effective anticancer with a specific target is urgently required. Streptomyces are widely known as the potential producers of new anticancer molecules. Previously reported that the methanol extract of Streptomyces sennicomposti GMY01 isolated from Krakal Coast, Gunungkidul had very strong cytotoxic activity against MCF-7 and T47D breast cancer cells with IC50 values of 0.6 and 1.3 µg/mL, respectively. The following study aimed to isolate and identify active compounds of the S. sennicomposti GMY01 and evaluate its cytotoxic activity. The study was started by re-culturing and re-fermented optimization of S. sennicomposti GMY01 in a larger volume, then the bacteria were extracted using methanol following the bioassay-guided isolation of the extract obtained. The active compounds obtained were then structurally determined using UV/Vis spectroscopy, Fourier Transform-Infrared (FT-IR), Liquid Chromatography-Mass Spectroscopy (LC-MS), 1H NMR, and 13C NMR and analyzed for their cytotoxic activity using MTT assay on MCF-7 and normal Vero cells line. The results showed that the culture of the S. sennicomposti GMY01 using Starch Nitrate Broth (SNB) media yields the best results compared to other culture media. An active anticancer compound namely mannotriose was successfully isolated from the methanol extract with an IC50 value of 5.6 µg/mL and 687 µg/mL against the MCF-7 and Vero cells lines, respectively, indicating that this compound showed strong cytotoxic activity with high selectivity.

Antibiofilm activity of marine microbial natural products: potential peptide- and polyketide-derived molecules from marine microbes toward targeting biofilm-forming pathogens.

Controlling and treating biofilm-related infections is challenging because of the widespread presence of multidrug-resistant microbes. Biofilm, a naturally occurring matrix of microbial aggregates, has developed intricate and diverse resistance mechanisms against many currently used antibiotics. This poses a significant problem, especially for human health, including clinically chronic infectious diseases. Thus, there is an urgent need to search for and develop new and more effective antibiotics. As the marine environment is recognized as a promising reservoir of new biologically active molecules with potential pharmacological properties, marine natural products, particularly those of microbial origin, have emerged as a promising source of antibiofilm agents. Marine microbes represent an untapped source of secondary metabolites with antimicrobial activity. Furthermore, marine natural products, owing to their self-defense mechanisms and adaptation to harsh conditions, encompass a wide range of chemical compounds, including peptides and polyketides, which are primarily found in microbes. These molecules can be exploited to provide novel and unique structures for developing alternative antibiotics as effective antibiofilm agents. This review focuses on the possible antibiofilm mechanism of these marine microbial molecules against biofilm-forming pathogens. It provides an overview of biofilm development, its recalcitrant mode of action, strategies for the development of antibiofilm agents, and their assessments. The review also revisits some selected peptides and polyketides from marine microbes reported between 2016 and 2023, highlighting their moderate and considerable antibiofilm activities. Moreover, their antibiofilm mechanisms, such as adhesion modulation/inhibition targeting biofilm-forming pathogens, quorum sensing intervention and inhibition, and extracellular polymeric substance disruption, are highlighted herein.

Flavonoid from Moringa oleifera leaves revisited: A review article on in vitro, in vivo, and in silico studies of antidiabetic insulin-resistant activity.

Diabetes mellitus (DM) occurs when the body experiences insulin deficiency or is unable to use insulin appropriately, which increases the blood glucose levels over the threshold. Moringa oleifera leaf is a widely used and scientifically proven herbal medicine to treat DM. The demand for the development of new drugs has prompted in vitro, in vivo, and in silico studies of antidiabetic insulin-resistant activity. This study aims to conduct a comprehensive study of the types of flavonoid and nonflavonoid compounds that have antidiabetic activity in insulin resistance mellitus using in vitro, in vivo, and in silico approaches. The literature review was conducted in accordance with the offered reporting items for systematic review. Major bibliographic databases, i.e. Scopus, PubMed, and DOAJ, covering original articles about the aforementioned issues between January 1, 2011 and December 31, 2021 were used. In this study, 274 articles were retrieved, of which 4 were duplicates, and after the titles were read, only 108 were left for analysis. After the abstract screening, 32 articles were eligible for the literature review. The results exhibit that flavonoids, including quercetin and kaempferol, and nonflavonoids, including anthraquinone, cytogluside (glycoside), hemlock tannin, phenolic steroid, and 2-phenylchromenylium (anthocyanins), have potential insulin-resistant antidiabetic activity in vitro, in vivo, and in silico. This has broadened the research into the development of new drugs.

Marine-Derived Streptomyces sennicomposti GMY01 with Anti-Plasmodial and Anticancer Activities: Genome Analysis, In Vitro Bioassay, Metabolite Profiling, and Molecular Docking.

To discover novel antimalarial and anticancer compounds, we carried out a genome analysis, bioassay, metabolite profiling, and molecular docking of marine sediment actinobacteria strain GMY01. The whole-genome sequence analysis revealed that Streptomyces sp. GMY01 (7.9 Mbp) is most similar to Streptomyces sennicomposti strain RCPT1-4T with an average nucleotide identity (ANI) and ANI based on BLAST+ (ANIb) values of 98.09 and 97.33% (>95%). An in vitro bioassay of the GMY01 bioactive on Plasmodium falciparum FCR3, cervical carcinoma of HeLa cell and lung carcinoma of HTB cells exhibited moderate activity (IC50 value of 46.06; 27.31 and 33.75 µg/mL) with low toxicity on Vero cells as a normal cell (IC50 value of 823.3 µg/mL). Metabolite profiling by LC-MS/MS analysis revealed that the active fraction of GMY01 contained carbohydrate-based compounds, C17H29NO14 (471.15880 Da) as a major compound (97.50%) and mannotriose (C18H32O16; 504.16903 Da, 1.96%) as a minor compound. Molecular docking analysis showed that mannotriose has a binding affinity on glutathione reductase (GR) and glutathione-S-transferase (GST) of P. falciparum and on autophagy proteins (mTORC1 and mTORC2) of cancer cells. Streptomyces sennicomposti GMY01 is a potential bacterium producing carbohydrate-based bioactive compounds with anti-plasmodial and anticancer activities and with low toxicity to normal cells.

Application of Response Surface Methodology (RSM) for the Optimization of Ultrasound-Assisted Extraction (UAE) of Moringa oleifera: Extraction Yield, Content of Bioactive Compounds, and Biological Effects In Vitro.

This study optimized ultrasound-assisted extraction conditions to maximize the extraction yield, total flavonoid content (TFC), total phenolic content (TPC), and DPP IV enzyme inhibitory activity from Moringa oleifera. The four UAE factors, solvent ratio (A), solvent-solid ratio (B), extraction temperature (C), and extraction time (D), were optimized using response surface methodology (RSM). A Box-Behnken design was used for the experimental design. The optimal conditions were found to be a 50% v/v solvent ratio, a 30% v/w solvent-solid ratio, 35 °C extraction temperature, and 45 min extraction time. The experimental value of extraction yield (R1), TFC (R2), TPC (R3), and DPP IV enzyme inhibitory activity (R4) (87.99% w/w, 56.63 mg QE/g extract, 97.26 mg GAE/g extract, and 93.32% inhibition, respectively) agreed with those predicted by RSM models (88.10% w/w, 56.61 mg QE/g extract, 97.16 mg GAE/g extract, and93.38% inhibition, respectively), thus demonstrating the appropriateness of the model used and the ability of the RSM to optimize the extraction conditions. Excellent DPP IV enzyme inhibitory activity was exhibited by M. oleifera compared with the standard, sitagliptin. While the modeled equation fits the data, the t-test is not significant, suggesting that the experimental values agree with those predicted by the RSM-BBD.

Quantification of Enhydrin and Uvedalin in the Ethanolic Extract of Smallanthus sonchifolius Leaves Validated by the HPLC Method.

Yacon leaf (Smallanthus sonchifolius, Asteraceae) ethanolic extracts are widely used in herbal medicine preparation for diabetes. They contain two sesquiterpene lactones (enhydrin (1) and uvedalin (2)) as major bioactive compounds. To provide a suitable method of analysis for the extract's quality control, we developed and validated a simultaneous HPLC-UV method using the compounds as markers. Compounds 1 and 2 were isolated using a freeze crystallization technique followed by a preparative HPLC. Spectrometry data for 1 and 2 were determined and compared to the literature. Chromatographic separation was carried out for 30 min with a mobile phase that used 60% water and 40% acetonitrile and a C18 column (250 × 4.6 mm, 5 µm) as the stationary phase. The flow was set to 1 mL min-1 and detection was conducted at 210 nm. The validation method was conducted according to the ICH guidelines, which included linearity, precision, accuracy, LOD, and LOQ. The calibration curve of both compounds was linear (R 2 > 0.9999), with the limit of detection and quantification as follows, respectively, 0.52 and 1.57 µg/mL for 1, and 0.144 and 0.436 µg/mL for 2. The percentages of recovery and repeatability (%RSD) were, 101.46 and 0.30% for 1, and 97.68 and 0.08% for 2, respectively. The 1 and 2 were 1.67 and 0.88% in the Ykal extract, and 1.26 and 0.56% in the Ycin extract, respectively. The method was found to be linear, precise, accurate, and suitable to be applied for control quality analyses of yacon leaf extract.

A Comprehensive Update on the Bioactive Compounds from Seagrasses.

Marine angiosperms produce a wide variety of secondary metabolites with unique structural features that have the potential to be developed as effective and potent drugs for various diseases. Recently, research trends in secondary metabolites have led to drug discovery with an emphasis on their pharmacological activity. Among marine angiosperms, seagrasses have been utilized for a variety of remedial purposes, such as treating fevers, mental disorders, wounds, skin diseases, muscle pain, and stomach problems. Hence, it is essential to study their bioactive metabolites, medical properties, and underlying mechanisms when considering their pharmacological activity. However, there is a scarcity of studies on the compilation of existing work on their pharmacological uses, pharmacological pathways, and bioactive compounds. This review aims to compile the pharmacological activities of numerous seagrass species, their secondary metabolites, pharmacological properties, and mechanism of action. In conclusion, this review highlights the potency of seagrasses as a promising source of natural therapeutical products for preventing or inhibiting human diseases.

Antibiofilm effect of C-10 massoia lactone toward polymicrobial oral biofilms.

This study is aimed to test the efficacy of C-10 Massoia lactone in oral polymicrobial degradation. Polymicrobial of Streptococcus sanguinis, Streptococcus mutans, Lactobacillus acidophilus, and Actinomyces viscosus were studied. C-10 Massoia lactone against biofilm degradation was investigated using modified crystal violet for biofilm staining. The effectiveness of C-10 Massoia lactone against biofilms was calculated by the minimum biofilm inhibitory concentration (MBIC50) and the minimum value of biofilm eradication concentration (MBEC50). Scanning electron microscope was used to study biofilm cell viability and morphological changes. The results showed a degradation effect of C-10 Massoia lactone against mature oral polymicrobial at 0.25% v/v. C-10 Massoia lactone can degrade polymicrobial biofilms of S. mutans, S. sanguinis, L. acidophilus and A. viscosus. This compound can destroy the extracellular polymeric substances (EPS) of polymicrobial biofilms. The potential application of C-10 Massoia lactone for anti-polymicrobial medication should be applied in such a way that any negative effects are minimized. Further research is needed to confirm the findings of this study.

Evaluation of the efficacy and toxicity of massoia oil nanoemulsion.

In order to enhance essential oil's stability and water insolubility, Massoia aromatica oil nanoemulsion was formulated and tested on the planktonic growth and biofilm formation of Pseudomonas aeruginosa, Staphylococcus aureus and Candida albicans; macrophage phagocytosis and on Vero cells viability. Oil in water nanoemulsion formula was optimized by using several solvents and co-solvents composition. The stability test of the formula was conducted by using a six cycle's freeze-thaw technique. Particle size and morphology were analyzed using a particle size analyzer and transmission electron microscopy. Microbial growth, biofilm formation inhibition, and cytotoxicity assays were performed on the optimized formula by using micro dilution methods. Mice macrophage phagocytosis activities against latex and C. albicans in the presence of samples were evaluated. Massoia nanoemulsion was obtained as a transparent yellowish emulsion having 99.6-99.9% of transmittance; physically and chemically stable; showed stronger antibacterial and antibiofilm on P. aeruginosa and S. aureus, moderate to C. albicans; no significant different on phagocytic activities. The IC50 of massoia oil nanoemulsion and massoia oil towards Vero cells were 35.9µg/mL and 107.5µg/mL respectively. Massoia oil nanoemulsion can protect the stability and decreases the hydrophobicity of the oil, conserve the antimicrobial and immunomodulatory activities, but increases its cytotoxicity.

In vivo Immunomodulatory Activity of Faloak Bark Extract (Sterculia quadrifida R.Br).

BACKGROUND AND OBJECTIVE: Faloak (Sterculia quadrifida R.Br) is widely used as traditional medicine in Indonesia to improve stamina (reduce tiredness for heavy workers). However, no scientific reports so far on the immunomodulatory effect. The aim of this study was to determine the effect of the bark of faloak as immunomodulatory agents by evaluating their effect on BALB/c mice lymphocytes proliferation, the activity of macrophage, nitric oxide production and the immunoglobulin G titer by in vivo techniques. MATERIALS AND METHODS: Decoction of the faloak bark was used for the in vivo assay. BALB/c mice were divided into 5 dose groups, each consisting of 5 mice. One group was chosen as the baseline, 3 groups were used for the group treated with the test substance at doses of 7.5, 11.75 and 17.5 g kg-1 of body weight of mice (p.o) and a positive control group was treated with Phyllanthus niruri Linn. (PN) extract (Stimuno®) 0.585 g kg-1 b.wt., (p.o). The test samples were given every day. All mice were induced by hepatitis B vaccine at day 7 and 14. The activity of in vivo assay was determined at day 19. The activity of immunomodulatory effect is expressed in phagocytic capacity, phagocytosis index, nitric oxide, OD of lymphocyte proliferation and IgG titers. RESULTS: The macrophage phagocytic capacity and phagocytosis index were significantly increased (p<0.05), nitric oxide production were altered significantly (p<0.05), but OD of lymphocyte proliferation and production of IgG titers were unchanged (p>0.05). CONCLUSION: This study showed that the faloak bark could increase the macrophages phagocytic activity, but no effect on lymphocyte cells and therefore did not influence the adaptive immune response.

Investigation on Inhibitory Potential of Myrmecodia tuberosa on Quorum Sensing-related Pathogenicity in Pseudomonas aeruginosa PAO1 and Staphylococcus aureus Cowan I Strains.

BACKGROUND AND OBJECTIVES: Tuber of Myrmecodia tuberosa Jack (Rubiaceae) has been widely used as herbal remedy in Indonesia. This research aimed to evaluate the effects of M. tuberosa Jack on quorum-sensing related pathogenicity of Pseudomonas aeruginosa and Staphylococcus aureus. MATERIALS AND METHODS: Following delipidation with petroleum ether, pulverized tuber was macerated in methanol. After evaporation to obtain methanol extract, liquid-liquid partition was perform on the extract to yield hexane, ethyl acetate and water fractions. The extract and fractions were screened for the effects on the planktonic growth of P. aeruginosa and S. aureus. Inhibition of pigment production was observed on cetrimide Agar following sample treatment on Pseudomonas aeruginosa. Samples were prepared as 2-0.0625 mg mL-1 concentration. The effects on swimming, swarming and twitching motility of Pseudomonas aeruginosa PAO1 following sample application were observed. All experiments were done in triplicate. RESULTS: Results showed that the ethyl acetate fraction caused a prominent effect on quorum sensing inhibition which might explain its biofilm inhibition effect on P. aeruginosa. Significant inhibitory effect in a concentration dependent manner towards pigment production inhibitor and motilities were observed over control. CONCLUSION: Despite being active as planktonic growth inhibitor towards S. aureus and P. aeruginosa, M. tuberosa ethyl acetate fraction is recommended to be investigated further as anti-infective against P. aeruginosa.

Effects of Costus speciosus ethanolic extract on male rats: The action mechanism and the ability to impregnate.

To examine the action mechanism that mediates the anti-fertility effect of Costus speciosus extract, research was conducted on male Sprague-Dawley rats. Costus extract was given to male rats for 14 days at various doses, namely 275, 550 and 1,100 mg kg-1 day-1 in 0.5% sodium CMC. The results showed that Costus extract with doses ranging from 275 to 1,100 mg kg-1 day-1 was able to inhibit pregnancy among female rats by 10-70%. No obstacles in terms of sexual behavior were identified among male rats. The anti-fertility effect of Costus extract kicked in without involving a decreased level of male reproductive hormones.

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.

Combinational effects of non n-Hexane Fractions of ant-plant (Myrmecodia tuberosa Jack) hypocotyl with doxorubicin against lymphocyte and cancer cells.

Doxorubicin is widely used as a chemotherapeutic drug despite having many side effects. It may cause the dysfunction of macrophage, decreasing proliferation of lymphocytes, decreasing CD4+/CD8+ ratio and inducing hepatotoxicity. Doxorubicin inhibits the growth of Vero, HeLa, and T47D cell lines, and also induces a resistance of MCF-7 cells. Previous studies showed that ethanolic extract and ethyl acetate fraction of ant-plant (Myrmecodia tuberose Jack) hipocotyl could increase macrophage phagocytosis activity and lymphocyte proliferation in vitro. Therefore, antplant is a potential immune stimulator. Combinational treatment of non n-hexane fraction (NHF) of ant-plant with doxorubicin did not affect the doxorubicin's potency. Nevertheless, increased lymphocyte viability induced by doxorubicin in varied dosages of NHF that lethal to HeLa, MCF-7 and T47D cells. Moreover, on Vero cells, doxorubicin became less toxic when induced together with NHF. Thus, NHF of ant-plant is potential to be proposed as doxorubicin co-chemotherapeutic agent against cancer cells.

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. SUMMARY: Massoia bark extracts (oily and solid phase) and C-10 Massoia lactone exhibited promising activities as antibiofilm against Candida albicans at IC50 are 0.074 %v/v, 271 µg/mL and 0.026 µg/mL respectively. The major constituent, C-10 Massoia lactone (C10H16O2) plays major role in exerting anticandida activity and potentially acts as an immunomodulator as well. However extracts showed inhibition of hyphae development of C. albicans which showed no correlation to the content of the Massoia lactone. Abbreviations used: GC/MS: Gas Chromatography/Mass Spectrometry, ML: Massoia Lactone, TLC: Thin Layer Chromatography, ATCC: American Type Culture Collection, RPMI: Roswell Park Memorial Institute, PBS: Phosphate Buffer Sterile, LSM: Laser scanning microscope, DMSO: Dimethyl sulfoxide, UV: Ultra violet, SDB: Sabouraud dextrose agar, MeOH: Methanol, LB: Luria Bertani, EtOAc: Ethyl acetate, CLSM: Confocal Laser Scanning Microscope, PI: Propidium iodide.

Pyrophen Produced by Endophytic Fungi Aspergillus sp Isolated from Piper crocatum Ruiz and Pav Exhibits Cytotoxic Activity and Induces S Phase Arrest in T47D Breast Cancer Cells.

Ethyl acetate extracts obtained from culture of endophytic fungi Aspergillus sp isolated from Piper crocatum Ruiz and Pav, have been shown to possess cytotoxic activity against T47D breast cancer cells. Investigations were here conducted to determine bioactive compounds responsible for the activity. Bioassay guided fractionation was employed to obtain active compounds. Structure elucidation was performed based on analysis of LC-MS, 1H-NMR, 13C-NMR, COSY, DEPT, HMQC, HMBC data. Cytotoxity assays were conducted in 96 well plates against T47D and Vero cell lines. Bioassay guided isolation and chemical investigation led to the isolation of pyrophen, a 4-methoxy-6-(1'-acetamido-2'-phenylethyl)-2H-pyran-2-one. Further analysis of its activity against T47D and Vero cells showed an ability to inhibit the growth of T47D cells with IC50 values of 9.2 µg/mL but less cytotoxicity to Vero cells with an IC50 of 109 µg/mL. This compound at a concentration of 400 ng/mL induced S-phase arrest in T47D cells.

Hydnophytum formicarum Jack ethanol extract modulates quorum sensing-controlled pathogenicity in Pseudomonas aeruginosa.

The discovery of new mechanism to control microbial pathogenicity by quorum sensing modulation has generated the search for quorum sensing inhibitor from natural resources. The objective of this research was to evaluate the ability of Hydnophytum formicarum Jack (Rubiaceae) ethanol extract to antagonize cell-to cell communication. Pulverized H. formicarum tuber was macerated in ethyl alcohol 96% and evaporated to yield ethanol extract. A dillution technique using Luria-Bertani (LB) medium was used to observe the capability of the extract to reduce the violacein production in Chromobacterium violaceum. Samples in two-fold dilution were prepared to obtain 2 - 0.0625 mg/mL concentration. The effects on swimming, swarming and twitching motility as well as the formation of biofilm towards Pseudomonas aeruginosa PAO1 were recorded over control. All experiments were done in triplicate. The architecture of Ps. aeruginosa biofilm treated with samples was examined by CLSM (Confocal Laser Scanning Microscopy) . Our results suggested that the ethanol extract of H. formicarum caused violacein production inhibition. Furthermore, inhibition of Ps. aeruginosa motility and biofilm formation were recorded to be significant over control in a concentration dependent manner. H. formicarum serves as a potential source for new QS-based antibacterial drugs towards Ps. aeruginosa.

Polysaccharide-Rich Fraction of Noni Fruit (Morinda citrifolia L.) as Doxorubicin Co-Chemotherapy: Evaluation of Catalase, Macrophages, and TCD8+ Lymphocytes.

Noni fruit (Morinda citrifolia L.) has been acknowledged for its cytotoxic and immunostimulatory activity. Our previous results on the immunomodulatory effect of a noni juice polysaccharide-rich fraction encouraged this research to evaluate the potency of the polysaccharide-rich fraction as co-chemotherapy with doxorubicin (DOX) administration. Macrophage activity (MA) was evaluated with the latex bead method. The phagocytic index (PI) was measured as the number of latex beads ingested by 100 macrophages, while the phagocytosis ratio (PR) was indicated by the percentage of macrophages that ingested three or more latex beads. The CEC was evaluated by using a commercial assay kit, while CD8+ T lymphocyte proliferation was evaluated using a flowcytometry method following in vivo administration. Thirty male Wistar rats were divided into five groups (n = 6 each). The control group received DOX via i.p. at a concentration of 4.67 mg/kg BW on days 1 and 4; four treatment groups received PF p.o. at a concentration of 25; 50; 100; 200 mg/kg BW daily, respectively, and additionally DOX i.p. 4.67 mg/kg BW (days 1 and 4) for 7 days. The phagocytic activity was not affected significantly by PF administration compared to the Dox control, but PF administration at a dose of 25 and 50 mg/kg BW has been proven to increase TCD8+ cell proliferation in combination with DOX. The catalase concentration, on the other hand, significantly decreased following PF administration at a dose of 100 mg/kg BW. The results suggest that the polysaccharide-rich fraction of noni juice might induce immunomodulatory effects via TCD8+ activation, have antioxidant activity, and thus might be a potential candidate to be used as an adjuvant to DOX chemotherapy.

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.

From anti-fouling to biofilm inhibition: new cytotoxic secondary metabolites from two Indonesian Agelas sponges.

Chemical investigation of Indonesian marine sponges Agelas linnaei and A. nakamurai afforded 24 alkaloid derivatives representing either bromopyrrole or diterpene alkaloids. A. linnaei yielded 16 bromopyrrole alkaloids including 11 new natural products with the latter exhibiting unusual functionalities. The new compounds include the first iodinated tyramine-unit bearing pyrrole alkaloids, agelanesins A-D. These compounds exhibited cytotoxic activity against L5178Y mouse lymphoma cells with IC(50) values between 9.25 and 16.76 muM. Further new compounds include taurine acid substituted bromopyrrole alkaloids and a new dibromophakellin derivative. A. nakamurai yielded eight alkaloids among them are three new natural products. The latter include the diterpene alkaloids (-)-agelasine D and its oxime derivative and the new bromopyrrole alkaloid longamide C. (-)-Agelasine D and its oxime derivative exhibited cytotoxicity against L5178Y mouse lymphoma cells (IC(50) 4.03 and 12.5 microM, respectively). Furthermore, both agelasine derivatives inhibited settling of larvae of Balanus improvisus in an anti-fouling bioassay and proved to be toxic to the larvae. (-)-Agelasine D inhibited the growth of planktonic forms of biofilm forming bacteria S. epidermidis (MIC<0.0877 microM) but did not inhibit biofilm formation whereas the oxime derivative showed the opposite activity profile and inhibited only biofilm formation but not bacterial growth. The structures of the isolated secondary metabolites were elucidated based on extensive spectroscopic analysis involving one- and two-dimensional NMR as well as mass spectrometry and comparison with literature data.