Targeted Delivery Prodigiosin to Choriocarcinoma by Peptide-Guided Dendrigraft Poly-l-lysines Nanoparticles.

The available and effective therapeutic means to treat choriocarcinoma is seriously lacking, mainly due to the toxic effects caused by chemotherapy and radiotherapy. Accordingly, we developed a method for targeting delivery of chemotherapeutical drugs only to cancer cells, not normal cells, in vivo, by using a synthetic placental chondroitin sulfate (CSA)-binding peptide (plCSA-BP) derived from malarial protein VAR2CSA. A 28 amino acids placental CSA-binding peptide (plCSA-BP) from the VAR2CSA was synthesized as a guiding peptide for tumor-targeting delivery, dendrigraft poly-L-lysines (DGL) was modified with plCSA-BP and served as a novel targeted delivery carrier. Choriocarcinoma was selected to test the effect of targeted delivery carrier, and prodigiosin isolated from Serratia marcescens subsp. lawsoniana was selected as a chemotherapeutical drug and encapsulated in the DGL modified by the plCSA-BP nanoparticles (DGL/CSA-PNPs). DGL/CSA-PNPs had a sustained slow-release feature at pH 7.4, which could specifically bind to the JEG3 cells and exhibited better anticancer activity than that of the controls. The DGL/CSA-PNPs induced the apoptosis of JEG3 cells through caspase-3 and the P53 signaling pathway. DGL/CSA-PNPs can be used as an excellent targeted delivery carrier for anticancer drugs, and the prodigiosin could be an alternative chemotherapeutical drug for choriocarcinoma.

Extended pulsated drug release from PLGA-based minirods.

The kinetics of degradation and sustained cancer drugs (paclitaxel (PT) and prodigiosin (PG)) release are presented for minirods (each with diameter of ~5 and ~6 mm thick). Drug release and degradation mechanisms were studied from solvent-casted cancer drug-based minirods under in vitro conditions in phosphate buffer solution (PBS) at a pH of 7.4. The immersed minirods were mechanically agitated at 60 revolutions per minute (rpm) under incubation at 37 °C throughout the period of the study. The kinetics of drug release was studied using ultraviolet visible spectrometry (UV-Vis). This was used to determine the amount of drug released at 535 nm for poly(lactic-co-glycolic acid) loaded with prodigiosin (PLGA-PG) samples, and at 210 nm, for paclitaxel-loaded samples (PLGA-PT). The degradation characteristics of PLGA-PG and PLGA-PT are elucidated using optical microscope as well as scanning electron microscope (SEM). Statistical analysis of drug release and degradation mechanisms of PLGA-based minirods were performed. The implications of the results are discussed for potential applications in implantable/degradable structures for multi-pulse cancer drug delivery.

A comparative study of the adhesion of biosynthesized gold and conjugated gold/prodigiosin nanoparticles to triple negative breast cancer cells.

This paper explores the adhesion of biosynthesized gold nanoparticles (AuNPs) and gold (Au) nanoparticle/prodigiosin (PG) drug nanoparticles to breast cancer cells (MDA-MB-231 cells). The AuNPs were synthesized in a record time (less than 30 s) from Nauclea latifolia leaf extracts, while the PG was produced via bacterial synthesis with Serratia marcescens sp. The size distributions and shapes of the resulting AuNPs were characterized using transmission electron microscopy (TEM), while the resulting hydrodynamic diameters and polydispersity indices were studied using dynamic light scattering (DLS). Atomic Force Microscopy (AFM) was used to study the adhesion between the synthesized gold nanoparticles (AuNPs)/LHRH-conjugated AuNPs and triple negative breast cancer cells (MDA-MB-231 cells), as well as the adhesion between LHRH-conjugated AuNP/PG drug and MDA-MB-231 breast cancer cells. The adhesion forces between LHRH-conjugated AuNPs and breast cancer cells are shown to be five times greater than those between AuNPs and normal breast cells. The increase in adhesion is shown to be due to the over-expression of LHRH receptors on the surfaces of MDA-MB-231 breast cancer cells, which was revealed by confocal immuno-fluorescence microscopy. The implications of the results are then discussed for the selective and specific targeting and treatment of triple negative breast cancer.

Towards an understanding of bacterial metabolites prodigiosin and violacein and their potential for use in commercial sunscreens.

OBJECTIVES: To exploit the microbial ecology of bacterial metabolite production and, specifically, to: (i) evaluate the potential use of the pigments prodigiosin and violacein as additives to commercial sunscreens for protection of human skin, and (ii) determine antioxidant and antimicrobial activities (against pathogenic bacteria) for these two pigments. METHODS: Prodigiosin and violacein were used to supplement extracts of Aloe vera leaf and Cucumis sativus (cucumber) fruit which are known to have photoprotective activity, as well as some commercial sunscreen preparations. For each, sunscreen protection factors (SPFs) were determined spectrophotometrically. Assays for antimicrobial activity were carried out using 96-well plates to quantify growth inhibition of Staphylococcus aureus and Escherichia coli. RESULTS: For the plant extracts, SPFs were increased by an order of magnitude (i.e. up to ~3.5) and those for the commercial sunscreens increased by 10-22% (for 4% w/w violacein) and 20-65% (for 4% w/w prodigiosin). The antioxidant activities of prodigiosin and violacein were approximately 30% and 20% those of ascorbic acid (a well-characterized, potent antioxidant). Violacein inhibited S. aureus (IC50 6.99 ± 0.146 µM) but not E. coli, whereas prodigiosin was effective against both of these bacteria (IC50 values were 0.68 ± 0.06 µM and 0.53 ± 0.03 µM, respectively). CONCLUSION: The bacterial pigments prodigiosin and violacein exhibited antioxidant and antimicrobial activities and were able to increase the SPF of commercial sunscreens as well as the extracts of the two plant species tested. These pigments have potential as ingredients for a new product range of and, indeed, represent a new paradigm for sunscreens that utilize substances of biological origin. We discussed the biotechnological potential of these bacterial metabolites for use in commercial sunscreens, and the need for studies of mammalian cells to determine safety.

Prodigiosins: a novel family of immunosuppressants with anti-cancer activity.

Prodigiosins (PrGs) are a family of promising therapeutic molecules, isolated mostly from Gram-negative bacteria and characterized by a common pyrryldipyrrylmethene structure with varying side chains. They show a broad spectrum of activities such as anti-microbial, anti-malarial, anti-cancer and immunosuppressive. PrGs are attracting increasing attention due to the ongoing research for less toxic, but effective agents for cancer chemotherapy and immunosuppression for preventing allograft rejection and autoimmunity. Different analogues have been synthesized and evaluated. This review discusses the immunosuppressive and anti-cancer activities of this class of compounds, as both involve inhibition of cell proliferation. The main focus is on the in vitro and in vivo immunosuppressive activity of the different PrGs and the mechanisms involved. PrGs primarily target the T cells, though some effects are observed on other cell types also. Unlike the well-known immunosuppressant cyclosporin A, PrGs do not inhibit the secretion of IL-2 but inhibit the mitogenic signaling from IL-2, suggesting a different mechanism of action. Janus tyrosine kinase 3 (Jak3) that associates with IL-2R upon activation is considered as the molecular target for PrGs. Its restricted expression makes Jak3 as an attractive target for immunosuppressive therapy. However, the available literature suggests that some other pathways are also influenced by the PrGs. These may be important for the anti-cancer activity, as well as immunosuppressive action. Therefore, PrGs appear to be potential candidates for pharmaceutical development as immunosuppressants and also as anti-cancer agents.

Differential genomic damage in different tumor lines induced by prodigiosin.

Prodigiosin is a secondary metabolite produced by Serratia marcercens. As this pigment is suggested to be a cancer drug, genotoxicity studies are necessary. The aim of the present investigation was to evaluate the genotoxic effects of prodigiosin on tumoral and normal cell lines, NCIH-292, MCF-7 and HL-60. A normal line BGMK was used as control. Genomic damage induced by prodigiosin was observed in all tumor lines as well as the control line. The pigment induced the formation of micronuclei in tumor cells. The present data confirm the antitumor potential of prodigiosin. However, these findings also raise concerns regarding its target-specific action, as genotoxic effects on normal cells also occurred.

Suppression of cytotoxic T cell induction in vivo by prodigiosin 25-C.

Prodigiosin 25-C (PrG25-C) was discovered as an immunosuppressant in the course of our screening for immunomodulating substances. In this system, PrG25-C inhibited T lymphocytes proliferation and was less suppressive against B lymphocytes. PrG25-C was also a powerful inhibitor of cytotoxic T cell induction by mixed lymphocyte reaction and completely suppressed induction of H-2 specific cytotoxic cells at 12.7 nM. PrG25-C also inhibited in vivo induction of H-2 restricted cytotoxic T lymphocytes at a dose of 0.5 mg/kg but had little myelotoxicity because numbers of blood leukocytes and splenocytes of PrG25-C-treated mice were comparable to those of nonsensitized mice. No inhibitory effects of PrG25-C were observed on the production of anti-SRBC antibody. These results indicate that PrG25-C is a T-lymphocyte-specific immunosuppressant.

Undecylprodigiosin selectively induces apoptosis in human breast carcinoma cells independent of p53.

Undecylprodigiosin (UP) is a bacterial bioactive metabolite produced by Streptomyces and Serratia. In this study, we explored the anticancer effect of UP. Human breast carcinoma cell lines BT-20, MCF-7, MDA-MB-231 and T47D and one nonmalignant human breast epithelial cell line, MCF-10A, were tested in this study. We found that UP exerted a potent cytotoxicity against all breast carcinoma cell lines in a dose- and time-dependent manner. In contrast, UP showed limited toxicity to MCF-10A cells, indicating UP's cytotoxic effect is selective for malignant cells. UP's cytotoxic effect was due to apoptosis, as confirmed by positive TUNEL signals, annexin V-binding, caspase 9 activation and PARP cleavage. Notably, UP-induced apoptosis was blocked by the pan-caspase inhibitor z-VAD.fmk, further indicating the involvement of caspase activity. Moreover, UP caused a marked decrease of the levels of antiapoptotic BCL-X(L), Survivin and XIAP while enhancing the levels of proapoptotic BIK, BIM, MCL-1S and NOXA, consequently favoring induction of apoptosis. Additionally, we found that cells with functional p53 (MCF-7, T47D) or mutant p53 (BT-20, MDA-MB-231) were both susceptible to UP's cytotoxicity. Importantly, UP was able to induce apoptosis in MCF-7 cells with p53 knockdown by RNA interference, confirming the dispensability of p53 in UP-induced apoptosis. Overall, our results establish that UP induces p53-independent apoptosis in breast carcinoma cells with no marked toxicity to nonmalignant cells, raising the possibility of its use as a new chemotherapeutic drug for breast cancer irrespective of p53 status.

[Effectiveness of treating chronic cholecystitis by prodigiozain electrophoresis]. / Ob éffektivnosti lechenii élektroforezom prodigiozana bol'nykh khronicheskim kholetsistitom

136 patients suffering from chronic cholecystitis were treated in the phase of exacerbation with antibiotics in combination with prodigiosan. It was found that inclusion of prodigiosan into the therapeutic complex had a positive effect on the immediate and late results of the treatment. Since intramuscular administration of prodigiosan was accompanied by a number of side effects, a method of the drug administration by means of electrophoresis on the area of the bile bladder was tested. It was shown that prodigiosan administered by the method of electrophoresis totally preserved its stimulating capacity with respect to the host. As with intramuscular administration of prodigiosan, the general reaction of the host and positive dynamics of the clinical and laboratory indices of the active process were observed and the content of lysozyme in the blood phagocytes reliably increased. The method of prodigiosan administration by means of electrophoresis was better tolerated by the patients and no side effects characteristic of the drug intramuscular administration were noted. When indicated, the use of prodigiosan by means of electrophoresis is recommended.

[Effect of prodigiosin and its combination with immunodepressants on the graft versus host reaction in mice]. / Vliianie prodigiozana i ego sochetaniia s immunodepressorami na reaktsiiu transplantat protiv khoziaina u myshei.

The local (lymph node) graft-versus-host reaction (GVHR) in F1 (CBA X C57BL/6) mice and the lethal GVHR in C57BL/6 mice were induced by transfer of lymph node cells of CBA mice with skin allotransplants from C57BL/6 mice. Prednisolone in combination with asathioprin (imuran) administered to CBA mice inhibited the GVHR. Prodigiosan used alone was not active, while in combination with immunodepressants it increased their inhibitory effect. Adhesive cells with a suppressive activity were detected in the spleen of mice treated with prodigiosan. Such cells were capable of suppressing the capacity of syngeneic lymphocytes for inducing the GVHR.

[Use of prodigioza in the overall treatment of chronic osteomyelitis]. / Primenenie prodigiozana v kompleksnom lechenii khronicheskogo osteomielita

Treatment of 188 patients suffering from chronic osteomyelitis of various etiology and localization with prodigiosan, a bacterial lipopolysaccharide favoured an increase in the nonspecific immunobiological reactivity: the phagocytic activity and leucocytic intensity increased, the average titers of iso- and heteroagglutinins, as well as the complementary activity of the serum became higher. The results of the clinico-laboratory studies are indicative of advisable use of prodigiosan in complex therapy of cases with chronic osteomyelitis.

[Study of the effect of tetracycline, prodigiosin and their combination on the dynamics of phagocytosis of plague bacteria]. / Izuchenie vliianiia tetratsiklina, prodigiozana i ikh sochetaniia na dinamiku fagotsitoza chumnykh bakterii

The advantage of the combined use of prodigiozan and tetracycline was observed in tissue culture of peritoneal macrophages of albino mice. Earlier digestion of the intracellular Y. pestis EV by the animal macrophages exposed to prodigiozan and treated with tetracycline was noted. It was shown that the macrophages preserved during several hours of cultivation in vitro their properties acquired in the animal organism under the effect of the substances administered.

[Effect of prodigiozan on the course of candidiasis of the oral mucosa clinically and in an experiment]. / Vliianie prodigiozana na techenie kandidoza slizistoi obolochki polosti rta v klinike i éksperimente

The effect of various doses of prodigiozan on Candida infection was studied under experimental and clinical conditions. Hamsters and guinea pigs were used in the experiments. Prodigiozan was administered intramuscularly to the hamsters in doses of 100 gamma/kg of the body weight 1 hour before the infection and intracutaneously to the guinea pigs in doses of 50 gamma/kg 2 days before the infection. The results showed that the drug accelerated the recovery of the animals. In complex therapy of a group of 22 patients with candidosis of the mouth mucosa of various localization prodigiozan was administered thrice in doses of 50 gamma/kg. This provided faster elimination of the pathological process.

[Effect of several antibiotics and their combination with prodigiozan on Y. pestis EV, engulfed by macrophages]. / Vliianie mekotorykh antibiotikov i ikh sochetanii s prodigiozanom na Y. pestis EV, pogloshchennye makrofagami.

The effect of various antibiotics, such as streptomycin, gentamicin, ampicillin, benzylpenicillin, tetracycline, chlortetracycline and levomycetin on the plague bacteria (strain Y. pestis EV) located inside the cells was studied. Peritoneal macrophages of albino mice with aceptic inflammation of the abdominal cavity caused by intraperitoneal administration of 2 ml of sterile meat-peptone broth were used in the experiments. The ratio of the macrophages and microbes was 1 : 50. A part of the mice were treated with prodigiozan 24 hours before taking the exudate. The preparations of the macrophages of albino mice with the microbes absorbed by them served as the control. The effect of the antibiotics and their combinations with prodigiozan was stimated by the coefficient of multiplication suppression against the control. The observations were made in dynamics. The studies showed that the macrophage activity of the mice treated with prodigiozan after exposure to the antibiotics was reliably higher than that in the control and digestion of the microbes located inside the cells started earlier, providing more complete phagocytosis.

[Effectiveness of prodigiozan in the therapy of chronic nonspecific diseases of the lungs]. / Effectivnost' prodigiozana v terapii bol'nykh s khronicheskimi nespetsificheskimi zabolevaniiami legkikh

The efficiency of the Soviet prodigiozan in therapy of chronic non-specific diseases of the lungs (18 cases with bronchial asthma and 26 cases with chronic pneumonia of the 1st and 2nd stages) in the phase of exacerbation was studied. The results of the treatment were estimated by the clinico-roentgenological pictures. The effect of prodigiozan on breathing and the functional state of the adrenal gland was studied in particular. Satisfactory results were obtained in the patients suffering from chronic pneumonia of the 1st and 2nd stages without clinical signs of the bronchial tree affection and in the patients with infection-allergic forms of bronchial asthma. Modest results were observed in the patients suffering from chronic pneumonia of the 1st and 2nd stages with the bronchial tree affection, especially when exacerbated by the asthma syndrome. In some patients the results of the treatment with prodigiozan were not stable. Prodigiozan improved the bronchial permeability, stimulated the sympatho-adrenal system and had no effect on the adrenal cortex.