Costus igneus, 'the Insulin plant' causing hypoglycaemia: Two case reports.

Costus igneus, commonly known as the 'Insulin plant' has been traditionally used in India to lower blood sugar levels. Its method of action is through a protein peculiar to itself, which has hypoglycaemic action. It therefore may prove harmful if used in excess, or together with other hypoglycaemic agents.

HPLC-ESI/MS-MS characterization of compounds in Dolomiaea costus extract and evaluation of cytotoxic and antiviral properties: molecular mechanisms underlying apoptosis-inducing effect on breast cancer.

BACKGROUND: Dolomiaea costus (syn: Saussurea costus; Family Asteraceae) occupies an important place in the traditional Chinese medicinal plants and is prescribed for a wide range of disorders. The current study aimed to tentatively identify the phytoconstituents of D. costus extract and to explore antiproliferative activity against human breast cancer cells and its possible apoptotic mechanism along with antiviral activity against human adenovirus 5 (Adv-5). METHODS: The phytoconstituents of 70% ethanol extract of D. costus were assessed using HPLC/ESI-MS/MS technique. The cell viability was investigated against breast cancer cell line (MCF-7) via 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Mechanistically, the apoptotic effects on the Bax, Bcl2 and Caspase 3 were determined via quantitative reverse transcriptase-polymerase chain reaction (RT-qPCR). Further, the antiviral activity was assessed against Adv-5 based on virucidal and adsorption mechanisms. RESULTS: The HPLC/MS analysis of the extract revealed tentative identification of twenty compounds of polyphenolic nature, mainly flavonoids, lignans, coumarins, and anthocyanidins. The plant extract showed a cytotoxic effect against MCF-7 and Vero cells with IC50 values of 15.50 and 44 µg/ml, respectively, indicating its aggressiveness against the proliferation of breast cancer cells as confirmed by apoptotic genes expression which revealed upregulation of Bax and Caspase 3 but further insight analysis is needed to explore exact mechanistic pathway. Antiviral activity against Adv-5 was observed at a non-toxic concentration of the tested extract. CONCLUSIONS: Such observations against human breast cancer and viral replication supported further studies for nanoformulations in drug delivery systems as targeting therapy and in vivo studies before biomedical applications.

Nephroprotective Effect of Costus (Saussurea costus) Ethanolic Extract on Oxaliplatin®-induced Nephrotoxicity in Adult Male Wistar Rats.

<b>Background and Objective:</b> Oxaliplatin^® is an antineoplastic platinum-based compound; nephrotoxicity is one of its most serious side effects. This study aimed to explore the nephroprotective potential of Costus Ethanolic Extract (CEE) against Oxaliplatin^®-induced nephrotoxicity. <b>Materials and Methods:</b> Adult male Wistar rats, weighting 140-160 g, were randomly divided into four groups: (1) Normal rats, (2) Rats ingested with CEE (67.08 mg kg¹ day¹), (3) Rats injected (ip) with Oxaliplatin^® (10 mg kg¹ week¹) and (4) rats treated with CEE in combination Oxaliplatin^® injection. <b>Results:</b> After six weeks of treatments, the results revealed that CEE ingestion along with Oxaliplatin^® injection markedly minimized the Oxaliplatin^®-induced renal deterioration; this was evidenced by the significant reduction in serum urea, creatinine, uric acid, Tumor Necrosis Factor Alpha (TNF-α), Interleukin 1Beta (IL¹ß) and Sodium ion (Na⁺) levels as well as kidney Malondialdehyde (MDA), Nitric Oxide (NO) and DNA fragmentation values. Controversially, a marked rise in serum Calcium, Potassium Ion (K⁺) and Cluster of Differentiation 4 (CD4) levels besides renal Glutathione (GSH), Catalase (CAT) and Superoxide Dismutase (SOD) values. Similarly, the histopathological findings confirmed the biochemical ones as the CEE restored the Oxaliplatin^®-induced histological degenerations. <b>Conclusion:</b> In conclusion, CEE exhibited nephron-protection efficiency against Oxaliplatin^®-induced nephrotoxicity; this promising effect may be achieved through the antioxidant and radical scavenging activities of its constituents.

Synthesis of ZnO nanoparticles using insulin-rich leaf extract: Anti-diabetic, antibiofilm and anti-oxidant properties.

Here, we report the novel fabrication of ZnO nanoparticles using the Costus igneus leaf extract. Gas chromatography-mass spectrometry (GC-MS) and proton nuclear magnetic resonance (1H NMR) spectroscopy to determine the bioactive components present in the plant extract. The synthesis of Ci-ZnO NPs (C. igneus- coated zinc oxide nanoparticles) was accomplished using a cost-effective and simple technique. Ci-ZnO NPs were specified using UV-visible spectroscopy, FTIR, XRD, and TEM. Ci-ZnO NPs was authenticated by UV-Vis and exhibited a peak at 365 nm. The XRD spectra proved the crystalline character of the Ci-ZnO NPs synthesized as hexagonal wurtzite. The FTIR spectrum illustrated the presence of possible functional groups present in Ci-ZnO NPs. The TEM micrograph showed evidence of the presence of a hexagonal organization with a size of 26.55 nm typical of Ci-ZnO NPs. The α-amylase and α-glucosidase inhibition assays demonstrated antidiabetic activity of Ci-ZnO NPs (74 % and 82 %, respectively), and the DPPH [2,2-diphenyl-1-picrylhydrazyl hydrate] assay demonstrated the antioxidant activity of the nanoparticles (75%) at a concentration of 100 µg/ml. The Ci-ZnO NPs exhibited promising antibacterial and biofilm inhibition activity against the pathogenic bacteria Streptococcus mutans, Lysinibacillus fusiformis, Proteus vulgaris, and Vibrio parahaemolyticus. Additionally, the Ci-ZnO NPs showed biocompatibility with mammalian RBCs with minimum hemolytic activity (0.633 % ±â€¯0.005 %) at a concentration of 200 µg/ml.

Raffinose from Costus speciosus attenuates lipid synthesis through modulation of PPARs/SREBP1c and improves insulin sensitivity through PI3K/AKT.

Among several metabolic disorders, the pathogenesis of insulin resistance is considered to be multifactorial. Raffinose, an oligosaccharide isolated from the rhizome of Costus speciosus showed ≤50% inhibition of lipid accumulation in differentiated HepG2 and 3T3-L1 cells through exhibiting partial agonism to PPARγ, and, an enhanced secretion of adiponectin in 3T3-L1 adipocytes. Raffinose was also observed to attenuate the expression of SREBP1c, ACC and FAS which are involved in the fatty acid synthesis. A corresponding upregulation of PPARα and ACO involved in fatty acid oxidation was observed in steatotic HepG2 hepatocytes and 3T3-L1 adipocytes. In vitro evaluation of its anti-diabetic potential showed a dose dependent enhancement of glucose uptake. Investigation of the insulin sensitizing efficacy of Raffinose revealed an increase in Glut4 translocation via phosphorylation of IRß/PI3K/Akt in differentiated L6 myocytes and 3T3-L1 preadipocytes. In addition, Raffinose was potentially involved in glycogen synthesis by inhibiting the activation of GSK3ß. Hence, Raffinose could be a useful therapeutic agent for metabolic maladies.

Comparative metabolic profiling of Costus speciosus leaves and rhizomes using NMR, GC-MS and UPLC/ESI-MS/MS.

Costus speciosus had been used in oriental systems of medicines, to treat diverse ailments. The present study was focused on NMR, GC-MS and UPLC/ESI-MS/MS-based metabolic profiling of C. speciosus. This metabolic study resulted in the identification of 91 and quantification of 69 metabolites. Caffeic acid derivatives previously unreported in C. speciosus were also identified. High quantity of steroidal saponins namely methyl protogracillin (297.97 ± 0.07 mg/g dried wt.) and dioscin (158.72 ± 0.27 mg/g dried wt.) were observed in butanol fraction of rhizomes. Health care metabolites including caffeic acid (37.88 ± 0.04 mg/g dried wt.) and trehalose (75.12 ± 0.08 mg/g dried wt.) were also detected in ethyl acetate and aqueous fractions of rhizomes, respectively. Metabolites of nutraceutical and biological significance including eremanthine (5.14 ± 0.68%, peak area), tocopherols (~22%), sterols (~25%) were also identified from hexane fractions of rhizomes and leaves using GC-MS. The analytical techniques used had successfully differentiated metabolites composition among leaves and rhizomes.

Growth kinetics and diosgenin estimation from callus cultures of Costus speciosus (Koen. ex. Retz.).

The present investigation reports the growth kinetics and diosgenin accumulation in callus cultures of Costus speciosus. Effect of explants, media and plant growth regulators was evaluated with respect to callus induction and growth. Out of the two explants viz pseudostem and seed, pseudostem showed maximum callus induction frequency of 90% on MS medium. The fresh weight of callus was maximum (9-folds) on 28th day on 1.0 mg/L picloram containing medium. The callus obtained was white compact hard (WCH). For growth kinetics study pseudostem derived callus was transferred on different media supplemented with 1.0 mg/L picloram. All phases of growth were seen in callus inoculated on all the three media except the absence of stationary phase on MS and SH media. MS medium proved to be the best for maximum biomass accumulation (9-fold) on 28th day of culture and callus in post-exponential phase showed maximum diosgenin accumulation (33 ppm).

Triagem fitoquímica e atividade antioxidante de Costus spicatus (Jacq. ) S. w / Phytochemical screening and antioxidant activity of Costus spicatus (Jacq. ) Sw

O objetivo do trabalho foi avaliar o perfil fitoquímico e a atividade antioxidante comparando-se os resultados obtidos entre diferentes órgãos da Costus spicatus. Foram utilizados caules, folhas e flores da Costus spicatus, colhidos na Universidade Estadual do Norte Fluminense Darcy Ribeiro, no município de Campos dos Goytacazes, RJ. Os diferentes órgãos foram avaliados quanto ao perfil fitoquímico e atividade antioxidante nas frações hexânica, acetato de etila e butanólica. Os resultados obtidos comprovaram a presença de Triterpenos e Esteróides (Lieberman-Burchard), Flavonóides (Shinoda), Saponinas (Índice de espuma), Alcalóides (Drangendorff) e Taninos (Cloreto Férrico). Foi demonstrada a atividade antioxidante potencial. Concluiu-se que a Costus spicatus apresentou todas as classes de metabólitos pesquisadas, dentre as quais, algumas possuem atividade biológica já conhecida, fazendo-se necessária a realização de estudos quantitativos e pesquisas que demonstrem seus efeitos farmacológicos, contribuindo para o desenvolvimento de novos fármacos.

The objective of this work was to evaluate the phytochemical profile and the antioxidant activity comparing the results between different organs of Costus spicatus. We used the stems, leaves and flowers of Costus spicatus harvested at the State University of Norte Fluminense "Darcy Ribeiro" in the city of Campos dos Goytacazes, Rio de Janeiro, Brazil. The different organs were evaluated for antioxidant activity and phytochemical profile in the hexane, ethyl acetate and butanol fractions. The results obtained proved the presence of triterpenes and steroids (Liebermann-Burchard), flavonoids (Shinoda), saponins (foaming index), alkaloids (Dragendorff) and tannins (ferric chloride). We demonstrated the potential antioxidant activity. We concluded that Costus spicatus presented all classes of metabolites studied, among which some already have their biological activity known by the literature, being necessary the performance of quantitative and research studies that demonstrate their pharmacological effects, thus contributing to the development of new drugs.

Orally active hypoglycemic protein from Costus igneus N. E. Br.: an in vitro and in vivo study.

Plants have been used for the treatment of diabetes since time immemorial. In the present study, insulin-like protein (ILP) is purified from Costus igneus belonging to family Costaceae from Western ghats of India. The ILP showed cross reactivity with murine anti-insulin antibodies hence was purified by affinity chromatography using anti-insulin antibodies. The characterization of ILP showed that it is structurally different from insulin but functionally similar. The ILP showed a hypoglycemic activity in an in vitro assay with insulin responsive cell line RIN 5f. Interestingly ILP showed significant decrease in blood glucose level when administered orally in oral glucose tolerance test. This was compared to insulin a positive control given intraperitoneally in streptozotocine induced diabetic mice. There was no toxic effect seen on animals after administrating the ILP. Therefore we conclude that the ILP purified in the present study from C. igneus is a novel protein having hypoglycemic activity.

Next generation sequencing and de novo transcriptome analysis of Costus pictus D. Don, a non-model plant with potent anti-diabetic properties.

BACKGROUND: Phyto-remedies for diabetic control are popular among patients with Type II Diabetes mellitus (DM), in addition to other diabetic control measures. A number of plant species are known to possess diabetic control properties. Costus pictus D. Don is popularly known as "Insulin Plant" in Southern India whose leaves have been reported to increase insulin pools in blood plasma. Next Generation Sequencing is employed as a powerful tool for identifying molecular signatures in the transcriptome related to physiological functions of plant tissues. We sequenced the leaf transcriptome of C. pictus using Illumina reversible dye terminator sequencing technology and used combination of bioinformatics tools for identifying transcripts related to anti-diabetic properties of C. pictus. RESULTS: A total of 55,006 transcripts were identified, of which 69.15% transcripts could be annotated. We identified transcripts related to pathways of bixin biosynthesis and geraniol and geranial biosynthesis as major transcripts from the class of isoprenoid secondary metabolites and validated the presence of putative norbixin methyltransferase, a precursor of Bixin. The transcripts encoding these terpenoids are known to be Peroxisome Proliferator-Activated Receptor (PPAR) agonists and anti-glycation agents. Sequential extraction and High Performance Liquid Chromatography (HPLC) confirmed the presence of bixin in C. pictus methanolic extracts. Another significant transcript identified in relation to anti-diabetic, anti-obesity and immuno-modulation is of Abscisic Acid biosynthetic pathway. We also report many other transcripts for the biosynthesis of antitumor, anti-oxidant and antimicrobial metabolites of C. pictus leaves. CONCLUSION: Solid molecular signatures (transcripts related to bixin, abscisic acid, and geranial and geraniol biosynthesis) for the anti-diabetic properties of C. pictus leaves and vital clues related to the other phytochemical functions like antitumor, anti-oxidant, immuno-modulatory, anti-microbial and anti-malarial properties through the secondary metabolite pathway annotations are reported. The data provided will be of immense help to researchers working in the treatment of DM using herbal therapies.

The centromeric heterochromatin of Costus spiralis: poorly methylated and transiently acetylated during meiosis.

The centromeric region of Costus spiralis is characteristically composed of a small heterochromatic DAPI(+) band flanked by a discrete decondensed region. High concentrations of serine 10 of histone H3 (H3S10ph) around the DAPI(+) band in pachytene chromosomes and the location of this heterochromatin at the chromosome region directed towards the poles during metaphase-anaphase I confirm its integration into the centromeric region. Antibodies against both typical components of euchromatin histones (histone H4 acetylated at lysine 5 (H4K5ac) and histone H3 dimethylated at lysine 4 (H3K4me2)) and heterochromatin (dimethylated lysine 9 of H3 (H3K9me2) and anti-5-methylcytosine (5-mC)) were used to characterize the centromeric chromatin of this species during meiosis. In pachytene chromosomes, the decondensed terminal euchromatin of the chromosome arms were seen to be richer in H4K5ac and H3K4me2 histones, while the more condensed proximal region was relatively stronger labeled with anti-H3K9me2 and anti-5-methylcytosine (5-mC). The centromeric region itself, including the DAPI(+) band, was poor in all of these chromatin modifications, but it was highly enriched in H4K5ac at pachytene. Before and after this stage, the centromeric region was poorly labeled with anti-H4K5ac. Hypomethylation and hyperacetylation of any kind of heterochromatin has rarely been reported, and it may be related to the dominant role of the centromere domain over the heterochromatin repeats.

Changes in expression pattern of the teosinte branched1-like genes in the Zingiberales provide a mechanism for evolutionary shifts in symmetry across the order.

PREMISE OF THE STUDY: Floral symmetry is a trait of key importance when considering floral diversification because it is thought to play a significant role in plant-pollinator interactions. The CYCLOIDEA/TEOSINTE BRANCHED1 (CYC/TB1)-like genes have been implicated in the development and evolution of floral symmetry in numerous lineages. We thus chose to investigate a possible role for these genes in the evolution of floral symmetry within petaloid monocots, using the order Zingiberales as a model system. In the Zingiberales, evolutionary shifts in symmetry have occurred in all floral whorls, making the order ideal for studying the evolution of this ecologically significant trait. METHODS: We analyzed TB1-like (TBL) genes from taxa spanning the order in a phylogenetic context. Using RNA in situ hybridization, we examined the expression of two TBL genes in Costus spicatus (Costaceae) and Heliconia stricta (Heliconiaceae), taxa with divergent floral symmetry patterns. KEY RESULTS: We identified Zingiberales-specific gene duplications as well as a duplication in the TBL gene lineage that predates the diversification of commelinid monocots. Shifts in TBL gene expression were associated with evolutionary shifts in floral symmetry and stamen abortion. ZinTBL1a expression was found in the posterior (adaxial) staminode of H. stricta and in the abaxial staminodial labellum of C. spicatus. ZinTBL2 expression was strongest in the anterior (abaxial) sepals of H. stricta and in the adaxial fertile stamen of C. spicatus. CONCLUSIONS: This study adds to the growing body of evidence that CYC/TB1-like genes have been repeatedly recruited throughout the course of evolution to generate bilateral floral symmetry (zygomorphy).

Different types of plant chromatin associated with modified histones H3 and H4 and methylated DNA.

Eukaryotic chromosomes are organized into two large and distinct domains, euchromatin and heterochromatin, which are cytologically characterized by different degrees of chromatin compaction during interphase/prophase and by post-synthesis modifications of histones and DNA methylation. Typically, heterochromatin remains condensed during the entire cell cycle whereas euchromatin is decondensed at interphase. However, a fraction of the euchromatin can also remain condensed during interphase and appears as early condensing prophase chromatin. 5S and 45S rDNA sites and telomere DNA were used to characterize these regions in metaphase and interphase nuclei. We investigated the chromosomal distribution of modified histones and methylated DNA in the early and late condensing prophase chromatin of two species with clear differentiation between these domains. Both species, Costus spiralis and Eleutherine bulbosa, additionally have a small amount of classical heterochromatin detected by CMA/DAPI staining. The distribution of H4 acetylated at lysine 5 (H4K5ac), H3 phosphorylated at serine 10 (H3S10ph), H3 dimethylated at lysine 4 or 9 (H3K4me2, H3K9me2), and 5-methylcytosine was compared in metaphase, prophase, and interphase cells by immunostaining with specific antibodies. In both species, the late condensing prophase chromatin was highly enriched in H4K5ac and H3K4me2 whereas the early condensing chromatin was very poor in these marks. H3K9me2 was apparently uniformly distributed along the chromosomes whereas the early condensing chromatin was slightly enriched in 5-methylcytosine. Signals of H3S10ph were restricted to the pericentromeric region of all chromosomes. Notably, none of these marks distinguished classical heterochromatin from the early condensing euchromatin. It is suggested that the early condensing chromatin is an intermediate type between classical heterochromatin and euchromatin.

Tamizaje fitoquímico, actividad analgésica y antiinflamatoria de decocción de Costus pictus D Don / Phytochemical screening, analgesic and antiinflammatory properties of Costus pictus D Don decoction

Introducción: Costus pictus D Don (caña mexicana) es una especie cultivada como ornamental y la decocción de hojas y tallos frescos se emplea tradicionalmente para afecciones urinarias, como infecciones, litiasis y cólicos renales en Cuba. Objetivos: identificar los principales grupos de metabolitos secundarios y validar la actividad analgésica y antiinflamatoria de la decocción de hojas y tallos frescos de C. pictus. Métodos: se identificaron los principales grupos de metabolitos secundarios presentes en la decocción mediante un tamizaje fitoquímico. Se evaluó en los modelos farmacológicos de granuloma inducido por algodón en ratas, retorcimientos inducidos por ácido acético 0,75 por ciento (0,1 mL/10 g intraperitoneal) y retirada de la cola inmersa en agua a 55 °C en ratones; las dosis administradas, por vía oral, fueron equivalentes a 0,5; 1 y 5 g de material vegetal fresco/kg de peso corporal. Resultados: la decocción mostró presencia de flavonoides, compuestos fenólicos, compuestos lactónicos y cumarinas, saponinas y azúcares reductores. Inhibió significativamente la respuesta dolorosa en ambos modelos, pero no tuvo efecto antiinflamatorio en el granuloma provocado por algodón. Conclusiones: los resultados contribuyen a validar el uso tradicional de la decocción de hojas y tallos frescos de C. pictus para aliviar el dolor por causa de afecciones renales

Introduction: Costus pictus D Don (Mexican reed) is a species for decorative purposes and the decoction of leaves and fresh stems is traditionally used for urinary disorders like infections, lithiasis and renal colic in Cuba. Objectives: to identify the main groups of secondary metabolites and to validate the analgesic and anti-inflammatory properties of a decoction of fresh leaves and stems from C. pictus. Methods: the main groups of secondary metabolites present in the decoction were identified by a phytochemical screening. The decoction was evaluated in pharmacological models of cotton-induced granulomas in rats; 0,75 por ciento acetic acid-induced cramps (0.1 mL/10 g intraperitoneal) and removal of the tail submerged into water at 55 °C in mice; the orally administered doses were 0.5; 1 and 5 g of fresh vegetable material/kg of body weight. Results: the decoction disclosed the presence of flavonoids, phenolic compounds, lactonic compounds and coumarins, saponins, and reducing sugars. It significantly reduced the painful response both models, but there was no antinflammatory effect on cotton-induced granuloma. Conclusions: the results contributed to validate the traditional use of C. pictus fresh leaves and stems in order to relieve the pain caused by renal diseases

Tamizaje fitoquímico, actividad analgésica y antiinflamatoria de decocción de Costus pictus D Don / Phytochemical screening, analgesic and antiinflammatory properties of Costus pictus D Don decoction

Introducción: Costus pictus D Don (caña mexicana) es una especie cultivada como ornamental y la decocción de hojas y tallos frescos se emplea tradicionalmente para afecciones urinarias, como infecciones, litiasis y cólicos renales en Cuba. Objetivos: identificar los principales grupos de metabolitos secundarios y validar la actividad analgésica y antiinflamatoria de la decocción de hojas y tallos frescos de C. pictus. Métodos: se identificaron los principales grupos de metabolitos secundarios presentes en la decocción mediante un tamizaje fitoquímico. Se evaluó en los modelos farmacológicos de granuloma inducido por algodón en ratas, retorcimientos inducidos por ácido acético 0,75 por ciento (0,1 mL/10 g intraperitoneal) y retirada de la cola inmersa en agua a 55 °C en ratones; las dosis administradas, por vía oral, fueron equivalentes a 0,5; 1 y 5 g de material vegetal fresco/kg de peso corporal. Resultados: la decocción mostró presencia de flavonoides, compuestos fenólicos, compuestos lactónicos y cumarinas, saponinas y azúcares reductores. Inhibió significativamente la respuesta dolorosa en ambos modelos, pero no tuvo efecto antiinflamatorio en el granuloma provocado por algodón. Conclusiones: los resultados contribuyen a validar el uso tradicional de la decocción de hojas y tallos frescos de C. pictus para aliviar el dolor por causa de afecciones renales(AU)

Introduction: Costus pictus D Don (Mexican reed) is a species for decorative purposes and the decoction of leaves and fresh stems is traditionally used for urinary disorders like infections, lithiasis and renal colic in Cuba. Objectives: to identify the main groups of secondary metabolites and to validate the analgesic and anti-inflammatory properties of a decoction of fresh leaves and stems from C. pictus. Methods: the main groups of secondary metabolites present in the decoction were identified by a phytochemical screening. The decoction was evaluated in pharmacological models of cotton-induced granulomas in rats; 0,75 por ciento acetic acid-induced cramps (0.1 mL/10 g intraperitoneal) and removal of the tail submerged into water at 55 °C in mice; the orally administered doses were 0.5; 1 and 5 g of fresh vegetable material/kg of body weight. Results: the decoction disclosed the presence of flavonoids, phenolic compounds, lactonic compounds and coumarins, saponins, and reducing sugars. It significantly reduced the painful response both models, but there was no antinflammatory effect on cotton-induced granuloma. Conclusions: the results contributed to validate the traditional use of C. pictus fresh leaves and stems in order to relieve the pain caused by renal diseases(AU)