Biogenic synthesis of gold nanoparticles using dual extract of tulsi-Vinca for breast cancer tumor regression in mice.

Aim: This work aims to synthesize the gold nanoparticles (GNPs) using a dual extract of tulsi and Vinca (T+V-Gold) for breast cancer tumor regression. Methods: The GNPs were synthesized and characterized for their microscopic, spectroscopic and crystalline properties. Further, the GNPs were investigated for in vitro and in vivo studies for the treatment of the 4T1-induced triple-negative breast cancer murine model. Results: The GNPs for 4T1 tumor-challenged mice resulted in delayed tumor development and lower tumor burden, with T+V-Gold demonstrating the highest prevention of tumor spread. The antitumor effect of T+V-Gold is highly significant in the glutathione family antioxidants glutathione S-transferase and glutathione in tumor tissue samples. Conclusion: The bioefficacy and anticancer outcomes of T+V-Gold nanoformulation can be used as therapeutic agents and drug-delivery vehicles.

Phytochemical Analysis and In Vitro Antileukemic Activity of Alkaloid-Enriched Extracts from Vinca sardoa (Stearn) Pignatti.

Vinca sardoa (Stearn) Pignatti, known as Sardinian periwinkle, is widely diffused in Sardinia (Italy). This species contains indole alkaloids, which are known to have a great variety of biological activities. This study investigated the antileukemic activity against a B lymphoblast cell line (SUP-B15) of V. sardoa alkaloid-rich extracts obtained from plants grown in Italy, in Iglesias (Sardinia) and Rome (Latium). All the extracts showed a good capacity to induce reductions in cell proliferation of up to 50% at the tested concentrations (1-15 µg/mL). Moreover, none of the extracts showed cytotoxicity on normal cells at all the studied concentrations.

Development of PSA@PS-TiO2 nanocomposite photocatalyst: structure, mechanism, and application using response surface designs and molecular modeling.

Using periwinkle shell ash (PSA) and polystyrene (PS), a new-fangled PSA@PS-TiO2 photocatalyst was fabricated. The morphological images of all the samples studied using a high-resolution transmission electron microscope (HR-TEM) showed a size distribution of 50-200 nm for all samples. The SEM-EDX showed that the membrane substrate of PS was well dispersed, confirming the presence of anatase/rutile phases of TiO2, and Ti and O2 were the major composites. Given the very rough surface morphology (atomic force microscopy (AFM)) due to PSA, the main crystal phases (XRD) of TiO2 (rutile and anatase), low bandgap (UVDRS), and beneficial functional groups (FTIR-ATR), the 2.5 wt.% of PSA@PS-TiO2 exhibited better photocatalytic efficiency for methyl orange degradation. The photocatalyst, pH, and initial concentration were investigated and the PSA@PS-TiO2 was reused for five cycles with the same efficiency. Regression modeling predicted 98% efficiency and computational modeling showed a nucleophilic initial attack initiated by a nitro group. Therefore, PSA@PS-TiO2 nanocomposite is an industrially promising photocatalyst for treating azo dyes, particularly, methyl orange from an aqueous solution.

Structural and vibrational investigations and molecular docking studies of a vinca alkoloid, vinorelbine.

Vinorelbine, a vinca alkaloid, is an antimitotic drug that inhibits polymerisation process of tubulins to microtubules, and is widely used in cancer chemotherapy. Due to the importance of the structure-activity relationship, in this work the conformational preferences of the vinorelbine molecule were surched by PM3 method. The obtained lowest energy conformer was then optimized at DFT/B3LYP/6-31G(d,p) level of theory and the structural characteristics were determined. Frontier orbital (HOMO, LUMO) and molecular electrostatic potential (MEP) analyses were performed for the optimized structure. The experimental FT-IR, Raman and UV-VIS spectral data of vinorelbine along with the theoretical DFT/B3LYP/6-31G(d,p) calculations were investigated in detail. The vibrational wavenumbers were assigned based on the calculated potential energy distribution (PED) of the vibrational modes. To shed light into the anticancer property of vinorelbine as microtubule destabilizer, the most favourable binding mode and the interaction details between vinorelbine and tubulin were revealed by molecular docking studies of vinorelbine into the α,ß-tubulin (PDB IDs: 4O2B; 1SA0; 7CNN) and binding free energies were calculated by the combination of Molecular Mechanics/Generalized Born Surface Area (MMGBSA) and Molecular Mechanics/Poisson-Boltzmann Surface Area (MM-PBSA) methods {MM/PB(GB)SA}. The calculated vinorelbine-7CNN binding free energy, using by MM/PB(GB)SA approach, was found to be the best (-50.39 kcal/mol), and followed by vinorelbine-4O2B (-28.5 kcal/mol) and vinorelbine-1SA0 (-17.59 kcal/mol) systems. Moreover, the interaction of vinorelbine with the cytochrome P450 enzymes (CYP), which are known to help in the metabolism of many drugs in the body, was investigated by docking studies against CYP2D6 and CYP3A4 targets.Communicated by Ramaswamy H. Sarma.

Agrobacterium-Mediated in Planta Transformation in Periwinkle.

Madagascar periwinkle (Catharanthus roseus, family Apocynaceae) is a reservoir of more than 130 monoterpene indole alkaloids (MIAs) including the famous anti-neoplastic dimeric MIAs vinblastine and vincristine, and anti-hypertensive monomeric MIAs ajmalicine and serpentine. Understanding the biosynthetic steps and regulatory factors leading to the formation of MIAs is crucial for rational engineering to achieve targeted enhancement of different MIAs. Due to its highly recalcitrant nature, C. roseus is considered genetically non-tractable for transformation at the whole-plant level. Though few reports have demonstrated tissue culture-mediated regeneration and transformation of C. roseus at whole-plant level recently, the efficiency and reproducibility of these protocols have been a major challenge. To overcome this, we have developed a tissue-culture-independent Agrobacterium-mediated in planta transformation method in C. roseus. Using this method, we were able to efficiently generate stable transgenic plants without relying on the cumbersome methods of tissue-culture regeneration and transformation. Moreover, the transformed plants obtained through this in planta method exhibited stability in subsequent generations. Our method is useful not only for the elucidation of biosynthetic and regulatory steps involved in MIA formation through transgenic plant approach but also for metabolic engineering at the whole-plant level in C. roseus.

Detecção de 'CA. Phytoplasma phoenicium' em plantas de pervinca (Cathranthus roseus) em Uttar Pradesh, Índia

Cathranthus roseus also known as periwinkle, an ornamental plant contains several medicinal values, was found with the symptoms of little leaf and witches' broom at Shahjahanpur location with the incidence of up to 8%. The phytoplasma etiology was confirmed through scanning electron microscopy examination in all the four-leaf samples. Molecular analysis through PCR with universal primer pairs P1/P6 followed by nested PCR with R16F2n/R16r2 primers yielded ~1.2kbp amplicons in all the four symptomatic leaf samples. One amplicon was eluted, purified, sequenced, and used in BLASTn searches, which showed maximum identity of periwinkle isolate with several isolates of 16SrIX group of phytoplasma. Further, phylogenic analysis and in silico RFLP confirmed the association of 16SrIX-C subgroup phytoplasma in little leaf and witches broom plants which is the first report from India.

Fluorometric quantification of alkaloids in the homeopathic mother tinctures of Vinca minor L. and Fumaria officinalis L.

To assess the toxic potential of the alkaloids, a quantification method is necessary. An ion pair extraction method was used for quantitative fluorometric determination of vincamine, protopine and all contained alkaloids in the mother tinctures of Vinca minor and Fumaria officinalis. The non-fluorescent alkaloids were transformed into an ion pair with sodium-9,10-dimethoxy-anthracene-sulfonate and then fluorometrically determined and quantified in this study. The applicable ion pair was extracted in a suitable organic solvent, where dichloromethane has proven to be beneficial. Conditions for the ion pairing and fluorometric quantification are given. The recovery rate was used to investigate the quality of determinability and the influence of the mother tincture matrix. The method was applied to determine the concentration of protopine in the range 0.1 - 15 µg/ml and of vincamine in the range of 0.5 - 20 µg/ml. The limit of detection was < 0.3 µg/ml, and the limit of quantification < 0.9 µg/ml for both alkaloids.

The Phytochemical Analysis of Vinca L. Species Leaf Extracts Is Correlated with the Antioxidant, Antibacterial, and Antitumor Effects.

The phytochemical analysis of Vinca minor, V. herbacea, V. major, and V. major var. variegata leaf extracts showed species-dependent antioxidant, antibacterial, and cytotoxic effects correlated with the identified phytoconstituents. Vincamine was present in V. minor, V. major, and V. major var. variegata, while V. minor had the richest alkaloid content, followed by V. herbacea. V. major var. variegata was richest in flavonoids and the highest total phenolic content was found in V. herbacea which also had elevated levels of rutin. Consequently, V. herbacea had the highest antioxidant activity followed by V. major var. variegata. Whereas, the lowest one was of V. major. The V. minor extract showed the most efficient inhibitory effect against both Staphylococcusaureus and E. coli. On the other hand, V. herbacea had a good anti-bacterial potential only against S. aureus, which was most affected at morphological levels, as indicated by scanning electron microscopy. The Vinca extracts acted in a dose-depended manner against HaCaT keratinocytes and A375 melanoma cells and moreover, with effects on the ultrastructure, nitric oxide concentration, and lactate dehydrogenase release. Therefore, the Vinca species could be exploited further for the development of alternative treatments in bacterial infections or as anticancer adjuvants.

Recovery of the salt marsh periwinkle (Littoraria irrorata) 9 years after the Deepwater Horizon oil spill: Size matters.

Prior studies indicated salt marsh periwinkles (Littoraria irrorata) were strongly impacted in heavily oiled marshes for at least 5 years following the Deepwater Horizon oil spill. Here, we detail longer-term effects and recovery over nine years. Our analysis found that neither density nor population size structure recovered at heavily oiled sites where snails were smaller and variability in size structure and density was increased. Total aboveground live plant biomass and stem density remained lower over time in heavily oiled marshes, and we speculate that the resulting more open canopy stimulated benthic microalgal production contributing to high spring periwinkle densities or that the lower stem density reduced the ability of subadults and small adults to escape predation. Our data indicate that periwinkle population recovery may take one to two decades after the oil spill at moderately oiled and heavily oiled sites, respectively.

Green Synthesis of Ag-MnO2 Nanoparticles using Chelidonium majus and Vinca minor Extracts and Their In Vitro Cytotoxicity.

Medicinal plants are often used as reducing agents to prepare metal nanoparticles through green-synthesis due to natural compounds and their potential as chemotherapeutic drugs. Thus, three types of eco-friendly Ag-MnO2 nanoparticles (Ag-MnO2NPs) were synthesized using C. majus (CmNPs), V. minor (VmNPs), and a 1:1 mixture of the two extracts (MNPs). These NPs were characterized using S/TEM, EDX, XRD, and FTIR methods, and their biological activity was assessed in vitro on normal keratinocytes (HaCaT) and skin melanoma cells (A375). All synthesized NPs had manganese oxide in the middle, and silver oxide and plant extract on the exterior. The NPs had different forms (polygonal, oval, and spherical), uniformly distributed, with crystalline structures and different sizes (9.3 nm for MNPs; 10 nm for VmNPs, and 32.4 nm for CmNPs). The best results were obtained with VmNPs, which reduced the viability of A375 cells up 38.8% and had a moderate cytotoxic effect on HaCaT (46.4%) at concentrations above 500 µg/mL. At the same concentrations, CmNPs had a rather proliferative effect, whereas MNPs negatively affected both cell lines. For the first time, this paper proved the synergistic action of the combined C. majus and V. minor extracts to form small and uniformly distributed Ag-MnO2NPs with high potential for selective treatments.

Anti-tubulin agents of natural origin: Targeting taxol, vinca, and colchicine binding domains.

Microtubules are a protein which is made of α- and ß-heterodimer. It is one of the main components of the cell which play a vital role in cell division especially in G2/M-phase. It exists in equilibrium dynamic of polymerization and depolymerization of α- and ß-heterodimer. It is one of the best targets for developing anti-cancer drugs. Various natural occurring molecules are well known for their anti-tubulin effect such as vinca, paclitaxel, combretastatin, colchicine etc. These microtubule-targeted drugs are acted through two processes (i) inhibiting depolymerization of tubulin (tubulin stabilizing agents) and (ii) inhibiting polymerization of tubulin (tubulin destabilizing agents). Now days, various binding domains have been explore through which these molecules are binding to tubulin but the three major binding domain of tubulin are taxol, vinca and colchicine binding domain. The present article mainly focus on the classification of various naturally occurring compounds on the basis of their inhibition processes (depolymerization and polymerization) and the site of interaction (targets taxol, vinca and colchicine binding domain) which has been hitherto reported. By placing all the naturally occurring taxol, vinca and colchicine binding site analogues at one place makes a better understanding of the tubulin interactions with known natural tubulin binders that would helps in the discovery of new and potent natural, semi-synthetic and synthetic analogues for treating cancer.

Catharanthus roseus (Apocynaceae) naturally infected with diverse phytoplasmas in Costa Rica / Catharanthus roseus (Apocynaceae) naturalmente infectada con diversos fitoplasmas en Costa Rica

Abstract Phytoplasmas (class Mollicutes) are causal agents of plant diseases with an economic impact on crops or threatening local biodiversity. A survey was conducted from 2012 to 2016 on infected Catharanthus roseus plants that exhibited symptoms reminiscent of phytoplasma infection throughout Costa Rica. A total of 73 plants were collected exhibiting symptoms such as virescence, phyllody, axillary proliferation, little leaf, leaf malformation, chlorosis, or yellowing. All samples were tested by nested PCR using phytoplasma universal and specific primer pairs. Phytoplasma infection was detected in 52 (71.2 %) of the plants collected. Phytoplasmas of six subgroups belonging to 16Sr groups I, III, IX, XIII and XV were identified based on sequencing and in silico RFLP analyses. 'Candidatus Phytoplasma asteris' (16SrI) was the predominant group among the positive samples (n = 30) showing variety of symptoms and wide distribution from sea level to ca. 1 400 m.a.s.l. in six of the seven Costa Rican provinces. Group 16SrIII was the second most abundant (14 samples); and the remaining three groups were seldom found in C. roseus (8 samples). Moreover, group 16SrXIII phytoplasma was detected for the first time in the country. To the best of our knowledge, this is the first report of natural infection of C. roseus with phytoplasma subgroups 16SrI-B, 16SrI-P, 16SrIII-F, 16SrIX-F, 16SrXIII-A, and 16SrXV-B in Costa Rica and Central America.

Resumen Los fitoplasmas (clase Mollicutes) son agentes causales de enfermedades de plantas que provocan pérdidas económicas o amenazan la biodiversidad local. Una recolecta de plantas de Catharanthus roseus que mostraban síntomas de posible infección con fitoplasmas se realizó en diferentes lugares de Costa Rica desde 2012 a 2016. Un total de 73 plantas fueron recolectadas con síntomas tales como viriscencia, filodia, brotación axilar múltiple, reducción foliar, deformación foliar, clorosis, y amarillamiento. Todas las muestras fueron evaluadas mediante PCR anidado usando los pares de imprimadores universales y específicos para fitoplasmas. Infección por fitoplasmas se detectó en 52 (71.2 %) de las muestras. Fitoplasmas de seis subgrupos dentro de los grupos 16Sr I, III, IX, XIII y XV fueron identificados basados en secuenciación del ADN y análisis de polimorfismos de restricción (RFLP) in silico. El grupo predominante encontrado en las muestras positivas (n = 30) fue el 16SrI ('CandidatusPhytoplasma asteris'), éste mostró variedad de síntomas y amplia distribución desde el nivel del mar hasta casi los 1 400 m.s.n.m. en seis de las siete provincias de Costa Rica. El grupo 16SrIII fue el segundo más abundante (14 muestras); y los restantes tres grupos se encontraron en pocas muestras de C. roseus (8 muestras). Además, fitoplasmas del grupo 16SrXIII se detectaron por primera vez en el país. De acuerdo a nuestro conocimiento, este es el primer informe de infección natural de C. roseus con fitoplasmas de los subgrupos 16SrI-B, 16SrI-P, 16SrIII-F, 16SrIX-F, 16SrXIII-A y 16SrXV-B en Costa Rica y Centroamérica.

[BIOLOGICALLY ACTIVE ALKALOIDS FROM RHIZOMES WITH ROOTS OF VINCA HERBACEA WALDST. ET KIT, GROWING IN GEORGIA].

Roots and rhizomes of Vinca herbacea Waldst. et Kit, were collected during early flowering and fruiting. Рhenophases biologically active substances I and II were obtained by liquid-liquid extraction. Dominant alkaloids: tabersonin, reserpine, maidine, norfluorocurarin and copsinin were obtained after the dispertion in citrare-phosfhate buffer and subsequent TLC. Accelerated restitution of granulocytopoiesis was observed in mice during both irradiation and myelotoxic drug-induced acute leucopenia. Increase in total WBC over 200% was observed after treatment by substance I in drug-induced leucopenia model (fivefold oral administration) and over 130% after treatment by substance I in irradiate mice (fivefold intraperitoneal administration). Morphological and anatomical structures of the underground organs of V. herbacea have been studied. The main microstructural characteristics are revealed - Rhizomes are characterized by coutinized epidermis, lamellar collenchyma, fibers and the texture of the vascular system of a monocyclic structure. The root system shows the whole cortex, the endoderm with Kaspar spots; the outer, radially continuous phloem tissue is located in the conducting system and distinguishes the cylindrical xylem tissue with annular and spiral-circular blood vessels.

Effects of Deepwater Horizon Oil on the Movement and Survival of Marsh Periwinkle Snails (Littoraria irrorata).

The Deepwater Horizon (DWH) oil spill resulted in the release of millions of barrels of oil into the Gulf of Mexico, and some marsh shorelines experienced heavy oiling including vegetation laid over under the weight of oil. Periwinkle snails (Littoraria irrorata) are a critical component of these impacted habitats, and population declines following oil spills, including DWH, have been documented. This study determined the effects of oil on marsh periwinkle movement and survivorship following exposure to oil. Snails were placed in chambers containing either unoiled or oiled laid over vegetation to represent a heavily impacted marsh habitat, with unoiled vertical structure at one end. In the first movement assay, snail movement to standing unoiled vegetation was significantly lower in oiled chambers (oil thickness ≈ 1 cm) compared to unoiled chambers, as the majority (∼75%) of snails in oiled habitats never reached standing unoiled vegetation after 72 h. In a second movement assay, there was no snail movement standing unoiled structure in chambers with oil thicknesses of 0.1 and 0.5 cm, while 73% of snails moved in unoiled chambers after 4h. A toxicity assay was then conducted by exposing snails to oil coated Spartina stems in chambers for periods up to 72 h, and mortality was monitored for 7 days post exposure. Snail survival decreased with increasing exposure time, and significant mortality (∼35%) was observed following an oil exposure of less than 24 h. Here, we have shown that oil impeded snail movement to clean habitat over a short distance and resulted in oil-exposure times that decreased survival. Taken together, along with declines documented by others in field surveys, these results suggest that marsh periwinkle snails may have been adversely affected following exposure to DWH oil.

Synthesis of LoganVir, a new carbocyclic nucleoside analogue.

Starting from a natural cyclopentanoid monoterpene belonging to the class of iridoid glucosides called loganin, we performed the synthesis of a new carbocyclic nucleoside, allowing the preparation of a new lead compound, with a potential HIV antiviral activity as an reverse transcriptase competitive inhibitor that we named LoganVir. The stereocontrol of the coupling reaction was completed utilizing the procedure described by Mitsunobu with a purinic base.

Three new monoterpenoid indole alkaloids from Vinca major.

Three new monoterpenoid indole alkaloids, 19-hydroxyl-10-methoxy-19, 20-dihydrovinorine (1), 19-O-acetyl-10-methoxy-19, 20-dihydrovinorine (2), and 19, 21α-dihydroxyl-10-methoxy-19, 20-dihydrovinorine (3), along with five known analogues (4-8), were isolated from the whole plants of Vinca major. The new structures were elucidated by extensive NMR and MS analysis and comparison with known compounds. In addition, compounds 1-3 were evaluated for their cytotoxicities against five human cancer cell lines.

Suppression of aggressive strains of 'Candidatus phytoplasma mali' by mild strains in Catharanthus roseus and Nicotiana occidentalis and indication of similar action in apple trees.

To study antagonistic interactions of 'Candidatus Phytoplasma mali' strains, graft inoculation of Catharanthus roseus and Nicotiana occidentalis was performed with mild strains 1/93Vin and 1/93Tab as suppressors and three aggressive strains as challengers. Inoculation of the suppressors was carried out in either the cross-protection modus prior to grafting of the challengers or by co-inoculating suppressors and challengers. Monitoring using multiplex real-time polymerase chain reaction assays revealed that, in long-term cross-protection trials with C. roseus, suppressor 1/93Vin was present in all root and randomly collected stem samples over the entire observation period. In contrast, the challengers were never detected in such stem samples and rarely in the roots. Following simultaneous inoculation, the suppressor successively colonized all stem and root regions whereas detection of challenger AT steadily decreased. However, this strain remained detectable in up to 13 and 27% of stem and root samples, respectively. The cross-protection trials with N. occidentalis yielded results similar to that of the cross-protection experiments with C. roseus. Comparison of the symptomatology of infected apple trees with the presence of putatively suppressive strains indicated that suppression of severe strains also occurs in apple. Phylogenetic analysis using a variable fragment of AAA+ ATPase gene AP460 of 'Ca. P. mali' revealed that suppressors 1/93Vin and 1/93Tab, together with several other mild strains maintained in apple, cluster distantly from obviously nonsuppressive strains that were predominantly highly virulent.

Pharmacological mechanisms of black cohosh in Sprague-Dawley rats.

BACKGROUND: Studies indicate that extracts and purified components from black cohosh inhibit the growth of human breast cancer cells, but the molecular targets and signaling pathways have not yet been defined. PURPOSE: This study examines the pharmacological mechanisms and toxicological effects in the short term of the herb black cohosh on female Sprague-Dawley rats. MATERIALS AND METHODS: To assess effects on gene activity and lipid content, we treated female Sprague-Dawley rats with an extract of black cohosh enriched in triterpene glycosides (27%) at 35.7 or 0mg/kg. Four animals for each group were sacrificed at 1, 6 and 24h after treatment; liver tissue and serum samples were obtained for gene expression and lipid analysis. RESULTS: Microarray analysis of rat liver tissue indicated that black cohosh markedly downregulated mitochondrial oxidative phosphorylation genes. Phospholipid biosynthesis and remodeling, PI3-Kinase and sphingosine signaling were upregulated, driven largely by an upregulation of several isoforms of phospholipase C. Hierarchical clustering indicated that black cohosh clustered with antiproliferative compounds, specifically tubulin binding vinca alkaloids and DNA alkylators. In support of this, black cohosh repressed the expression of cyclin D1 and ID3, and inhibited the proliferation of HepG2, p53 positive, liver cancer cells. Black cohosh reduced the level of free fatty acids at 6 and 24h and triglycerides at 6h in the serum, but increased the free fatty acid and triglyceride content of the treated livers at 24h. CONCLUSION: Our results suggest that black cohosh warrants further study for breast cancer prevention and therapy.

[Thermodynamics of calmodulin and tubulin binding to the vinca-alkaloid vinorelbine].

Vinca-alkaloids, such as vinblastine, and some of their derivatives, as for example vinorelbine, are widely used in clinical therapy of leukemia and several types of tumors. Their effects are associated with the disfunctioning of the mitotic spindle, which leads to mitosis blockage and a shutdown of the cell cycle. Their primary target is tubulin, however recent research has shown that some of the vinca-alkaloids inhibit calmodulin binding to its targets. Vinka-alkaloids binding with other proteins could be responsible for their efficiency and neuroprotection. Here we investigated the thermodynamics of vinorelbine interactions with calmodulin and tubulin. It was determined that unlike the other vinca-alkaloids both vinorelbine binding sites are located in the C-domain of calmodulin, and characterized by association constants of 4.0 x 10(5) and 5.4 x 10(4) M(-1). At the same time the thermodynamics of vinorelbine binding to tubulin are not much different from that of other vinca-alkaloids. These results will allow getting a better insight on the reaction mechanisms of vinca-alkaloids on a secondary protein target.