Pyrogallol from Spirogyra neglecta Inhibits Proliferation and Promotes Apoptosis in Castration-Resistant Prostate Cancer Cells via Modulating Akt/GSK-3ß/ß-catenin Signaling Pathway.

Castration-resistant prostate cancer (CRPC) is an advanced form of prostate cancer associated with poor survival rates. The high proliferation and metastasis rates have made CRPC one of the most challenging types of cancer for medical practitioners and researchers. In this study, the anti-cancer properties and inhibition of CRPC progression by S. neglecta extract and its active constituents were determined using two CRPC cell lines, DU145 and PC3. The ethyl acetate fraction of S. neglecta (SnEA) was obtained using a solvent-partitioned extraction technique. The active constituents of SnEA were then determined using the HPLC technique, which showed that SnEA mainly contained syringic acid, pyrogallol, and p-coumaric acid phenolic compounds. After the determination of cytotoxic properties using the SRB assay, it was found that pyrogallol, but not the other two major compounds of SnEA, displayed promising anti-cancer properties in both CRPC cell lines. SnEA and pyrogallol were then further investigated for their anti-proliferation and apoptotic induction properties using propidium iodide and Annexin V staining. The results showed that SnEA and pyrogallol inhibited both DU145 and PC3 cell proliferation by inducing cell cycle arrest in the G0/G1 phase and significantly decreased the expression of cell cycle regulator proteins (cyclin D1, cyclin E1, CDK-2, and CDK-4, p < 0.001). SnEA and pyrogallol treatments also promoted apoptosis in both types of CRPC cells through significantly downregulating anti-apoptotic proteins (survivin, Bcl-2, and Bcl-xl, p < 0.001) and upregulating apoptotic proteins (cleaved-caspase-9, cleaved-caspase-3 and cleaved-PARP-1, p < 0.001). Mechanistic study demonstrated that SnEA and pyrogallol inactivated the Akt signaling pathway leading to enhancement of the active form of GSK-3ß in CRPC cell lines. Therefore, the phosphorylation of ß-catenin was increased, which caused degradation of the protein, resulting in a downregulation of ß-catenin (unphosphorylated form) transcriptional factor activity. The current results reflect the potential impact of S. neglecta extract and pyrogallol on the management of castration-resistant prostate cancer.

Heat stress response in the closest algal relatives of land plants reveals conserved stress signaling circuits.

All land plants (embryophytes) share a common ancestor that likely evolved from a filamentous freshwater alga. Elucidating the transition from algae to embryophytes - and the eventual conquering of Earth's surface - is one of the most fundamental questions in plant evolutionary biology. Here, we investigated one of the organismal properties that might have enabled this transition: resistance to drastic temperature shifts. We explored the effect of heat stress in Mougeotia and Spirogyra, two representatives of Zygnematophyceae - the closest known algal sister lineage to land plants. Heat stress induced pronounced phenotypic alterations in their plastids, and high-performance liquid chromatography-tandem mass spectroscopy-based profiling of 565 transitions for the analysis of main central metabolites revealed significant shifts in 43 compounds. We also analyzed the global differential gene expression responses triggered by heat, generating 92.8 Gbp of sequence data and assembling a combined set of 8905 well-expressed genes. Each organism had its own distinct gene expression profile; less than one-half of their shared genes showed concordant gene expression trends. We nevertheless detected common signature responses to heat such as elevated transcript levels for molecular chaperones, thylakoid components, and - corroborating our metabolomic data - amino acid metabolism. We also uncovered the heat-stress responsiveness of genes for phosphorelay-based signal transduction that links environmental cues, calcium signatures and plastid biology. Our data allow us to infer the molecular heat stress response that the earliest land plants might have used when facing the rapidly shifting temperature conditions of the terrestrial habitat.

Heavy Metals Assessment in Water, Sediments, Algae and Two Fish Species from River Swat, Pakistan.

The heavy metals were studied in water, sediments, algae, and various tissues of Glyptosternon reticulatum and Cyprinus carpio from River Swat, Pakistan, using flame atomic absorption spectrophotometer. The Zn, Cu, Pb and Ni were higher in water at sewage site compared to upstream and downstream sites. In sediments, the Ni and Cd were not detected whereas Cu, Pb and Zn were higher at downstream followed by sewage and upstream sites. The Ni and Zn in algae were higher at upstream and sewage sites compared to downstream site whereas Pb and Cd were higher at upstream site compared to sewage and downstream sites and Cu was found same at all the three sites. The heavy metals (Zn > Cu > Pb and Ni) in tissues (liver > gills > skin > muscles) of G. reticulatum was higher than in C. carpio. This study recommends the proper monitoring of River Swat in order to save its water and inhabitant aquatic life.

Removal of copper from an electroplating industrial effluent using the native and modified spirogyra.

In the present study, biosorption behavior of a green filamentous alga, spirogyra in its native and modified states was investigated for copper removal from an electroplating industrial effluent. For this, the effluent containing 194 mg·L-1 Cu2+ in sulfate medium was contacted with both forms of spirogyra, under the parametric variations of effluent pH, adsorbent dosage, contact time, and sorption temperature. The study revealed spirogyra as a prominent candidate for removing contaminant metal cation; however, at the same condition, biosorption capacity of modified biomass in gel form was higher than the native spirogyra. At the optimized condition with 6 g sorbent dosage treated to 100 mL effluent for 30 min at pH 6.0 and temperature 20 °C, the maximum 82.8% and 96.4% copper could be adsorbed by the native and modified spirogyra, respectively. The batch sorption data using native biomass followed pseudo-first-order kinetic; exhibiting the multilayer sorption mechanism via surface diffusion could be defined by the Freundlich model. In contrast, the sulfuric acid treated modified spirogyra followed pseudo-second-order kinetics and intra particle diffusion as the rate-limiting step.

Transcriptome Profiling of the Green Alga Spirogyra pratensis (Charophyta) Suggests an Ancestral Role for Ethylene in Cell Wall Metabolism, Photosynthesis, and Abiotic Stress Responses.

It is well known that ethylene regulates a diverse set of developmental and stress-related processes in angiosperms, yet its roles in early-diverging embryophytes and algae are poorly understood. Recently, it was shown that ethylene functions as a hormone in the charophyte green alga Spirogyra pratensis Since land plants evolved from charophytes, this implies conservation of ethylene as a hormone in green plants for at least 450 million years. However, the physiological role of ethylene in charophyte algae has remained unknown. To gain insight into ethylene responses in Spirogyra, we used mRNA sequencing to measure changes in gene expression over time in Spirogyra filaments in response to an ethylene treatment. Our analyses show that at the transcriptional level, ethylene predominantly regulates three processes in Spirogyra: (1) modification of the cell wall matrix by expansins and xyloglucan endotransglucosylases/hydrolases, (2) down-regulation of chlorophyll biosynthesis and photosynthesis, and (3) activation of abiotic stress responses. We confirmed that the photosynthetic capacity and chlorophyll content were reduced by an ethylene treatment and that several abiotic stress conditions could stimulate cell elongation in an ethylene-dependent manner. We also found that the Spirogyra transcriptome harbors only 10 ethylene-responsive transcription factor (ERF) homologs, several of which are regulated by ethylene. These results provide an initial understanding of the hormonal responses induced by ethylene in Spirogyra and help to reconstruct the role of ethylene in ancestral charophytes prior to the origin of land plants.

Heavy metal bioaccumulation in sediment, common reed, algae, and blood worm from the Shoor river, Iran.

Concentrations of 11 metals (cadmium, zinc, copper (Cu), vanadium (V), lead, magnesium (Mg), manganese, aluminum, iron (Fe), chromium (Cr), and nickel), and one metalloid (arsenic (As)) were measured in sediment, common reed (Phragmites australis), algae (Spirogyra sp.), and blood worm (Chironomus sp.) tissues of samples collected from the Shoor river. Samples were dried, acid digested, and the concentrations of metals were measured using inductively coupled plasma-optical emission spectrometer. A higher concentration of heavy metals was accumulated in Spirogyra and Chironomids than sediment and common reed. The highest rate of accumulation was found for Mg, V, Fe, As, Cu, and Cr. Spirogyra and Chironomids are capable of accumulating and thereby removing metals from polluted water bodies and are suitable for biomonitoring purposes.

Evidence for land plant cell wall biosynthetic mechanisms in charophyte green algae.

BACKGROUND AND AIMS: The charophyte green algae (CGA) are thought to be the closest living relatives to the land plants, and ancestral CGA were unique in giving rise to the land plant lineage. The cell wall has been suggested to be a defining structure that enabled the green algal ancestor to colonize land. These cell walls provide support and protection, are a source of signalling molecules, and provide developmental cues for cell differentiation and elongation. The cell wall of land plants is a highly complex fibre composite, characterized by cellulose cross-linked by non-cellulosic polysaccharides, such as xyloglucan, embedded in a matrix of pectic polysaccharides. How the land plant cell wall evolved is currently unknown: early-divergent chlorophyte and prasinophyte algae genomes contain a low number of glycosyl transferases (GTs), while land plants contain hundreds. The number of GTs in CGA is currently unknown, as no genomes are available, so this study sought to give insight into the evolution of the biosynthetic machinery of CGA through an analysis of available transcriptomes. METHODS: Available CGA transcriptomes were mined for cell wall biosynthesis GTs and compared with GTs characterized in land plants. In addition, gene cloning was employed in two cases to answer important evolutionary questions. KEY RESULTS: Genetic evidence was obtained indicating that many of the most important core cell wall polysaccharides have their evolutionary origins in the CGA, including cellulose, mannan, xyloglucan, xylan and pectin, as well as arabino-galactan protein. Moreover, two putative cellulose synthase-like D family genes (CSLDs) from the CGA species Coleochaete orbicularis and a fragment of a putative CSLA/K-like sequence from a CGA Spirogyra species were cloned, providing the first evidence that all the cellulose synthase/-like genes present in early-divergent land plants were already present in CGA. CONCLUSIONS: The results provide new insights into the evolution of cell walls and support the notion that the CGA were pre-adapted to life on land by virtue of the their cell wall biosynthetic capacity. These findings are highly significant for understanding plant cell wall evolution as they imply that some features of land plant cell walls evolved prior to the transition to land, rather than having evolved as a result of selection pressures inherent in this transition.

Proteomic analysis of Spirogyra varians mutant with high starch content and growth rate induced by gamma irradiation.

This study was conducted to develop a high-efficiency strain of Spirogyra varians for the production of biomass by radiation breeding. The characteristics of wild-type and mutant S. varians were analyzed through phenomenological and proteomic observations. The results of our phenomenological observations of the S. varians mutant demonstrated increases in growth rate and content of chlorophyll a, b, and a + b; in particular, a significant threefold increase was observed in starch accumulation. Proteomic analysis to investigate the differences in expression between wild-type and mutant proteins identified 18 proteins with significantly different expressions. From the literature review, it was confirmed that the up-regulated proteins were mainly involved in photosynthesis, carbohydrate biosynthesis, and energy metabolism. These results suggest the possibility of algae development by radiation breeding for the production of biofuel.

Combined in vitro and in vivo investigation of barite microcrystals in Spirogyra (Zygnematophyceae, Charophyta).

We have investigated the biomineralisation of barite ‒a useful proxy for reconstructing paleoproductivity‒ in a freshwater alga, Spirogyra, by combining in vitro and in vivo approaches to unveil the nature of its barite microcrystals. Scanning electron microscope (SEM) and energy-dispersive X-ray spectroscopy (EDXS) observations on simply dried samples revealed that the number and size of barite crystals were related to the barium concentration in the media. Additionally, their morphology showed a crystallographic face (011), which is not normally observed, suggesting the influence of organic molecules on the growth kinetics. The critical point drying method was used to preserve the internal and external structures of Spirogyra cells for SEM imaging. Crystals were found adjacent to the cytoplasmic membrane, near chloroplasts and fibrillary network. In vivo optical microscopy and Raman tweezer microspectroscopy in living cells showed that barite microcrystals are optically visible and follow cytoplasmic streaming. These results led us to propose that barite formation in Spirogyra occurs in the cytoplasm where barium and sulphate are both available: barium supplied non-selectively through the active transport of the divalent cations needed for actin polymerisation, and sulphate because necessary for amino acid biosynthesis in chloroplasts.

Algae utilization in assessment of the large Turawa Lake (Poland) pollution with heavy metals.

This investigation was undertaken to determine the applicability of algae for the assessment of contamination level of water reservoirs with heavy metals. The alga Spirogyra sp. collected in the littoral zone of the Large Turawa Lake (artificial lake in Southern Poland) was used for the study. The concentrations of heavy metals accumulated in the alga inhabiting a flow-through water basin of the Large Turawa Lake were found to correlate with sources of these metals, such as benthic sediments and contaminated watercourses. The highest concentrations of metals were found in alga samples collected at the outlet of the lake: c (Mn) = 12330 mg/kg dry mass, c (Fe) = 15059 mg/kg d.m., c (Cu) = 47.5 mg/kg d.m., c (Zn) = 1411 mg/kg d.m., c (Cd) = 108.8 mg/kg d.m., and c (Pb) = 684 mg/kg d.m. The metals originated from benthic sediments (sapropelic mud) deposited close to the outlet of the lake. Statistically significant differences in the concentrations of cadmium accumulated in the alga were found between samples from the sites, where cadmium occurred in sandy sediments (max. 27.6 mg/kg d.m.), and samples from the sites located far from the contaminated sediments (max. 12.8 mg/kg d.m.).

Enhancement of fermentative hydrogen production from Spirogyra sp. by increased carbohydrate accumulation and selection of the biomass pretreatment under a biorefinery model.

In this work, hydrogen (H2) was produced through the fermentation of Spirogyra sp. biomass by Clostridium butyricum DSM 10702. Macronutrient stress was applied to increase the carbohydrate content in Spirogyra, and a 36% (w/w) accumulation of carbohydrates was reached by nitrogen depletion. The use of wet microalga as fermentable substrate was compared with physically and chemically treated biomass for increased carbohydrate solubilisation. The combination of drying, bead beating and mild acid hydrolysis produced a saccharification yield of 90.3% (w/w). The H2 production from Spirogyra hydrolysate was 3.9 L H2 L-1, equivalent to 146.3 mL H2 g-1 microalga dry weight. The presence of protein (23.2 ± 0.3% w/w) and valuable pigments, such as astaxanthin (38.8% of the total pigment content), makes this microalga suitable to be used simultaneously in both food and feed applications. In a Spirogyra based biorefinery, the potential energy production and food-grade protein and pigments revenue per cubic meter of microalga culture per year was estimated on 7.4 MJ, US $412 and US $15, respectively, thereby contributing to the cost efficiency and sustainability of the whole bioconversion process.

Cancer chemopreventive effect of Spirogyra neglecta (Hassall) Kützing on diethylnitrosamine-induced hepatocarcinogenesis in rats.

Spirogyra neglecta, a freshwater green alga, is a local food in the northern and northeastern parts of Thailand. This investigation explored the anticarcinogenicity of S neglecta and its possible cancer chemopreventive mechanisms in rats divided into 14 groups. Groups 1 and 10 served as positive and negative control groups, respectively. Groups 1-9 were intraperitoneally injected with diethylnitrosamine (DEN) once a week for 3 weeks. Groups 10-14 received normal saline instead. One week after the last DEN injection, groups 2-5 were administered for 9 consecutive weeks various doses of S neglecta extract (SNE) and dried S neglecta (SND), mixed with basal diet. Groups 6-9 and 11-14 similarly were administered various doses of SNE and SND starting from the first week of the experiment. Administration of SNE and SND was not associated with formation of glutathione-S- transferase placental form (GST-P) positive foci in rat liver. SNE and SND during initiation phase significantly reduced the number of GST-P positive foci in rats injected with DEN. The number of GST-P also diminished in groups treated with SNE and SND after injection with DEN, except for the low dose extract group. SNE showed stronger anticarcinogenic potency than SND. Furthermore, SNE also decreased the number of Ki-67 positive cells. However, the numbers of TUNEL-positive cells in the liver of the SNE-treated groups were not statistically different from the controls. The GST activity in 50 mg/kg bw of SNE and 1% of SND groups was significantly increased as compared to the positive control. In conclusion, Spirogyra neglecta (Hassall) Kutzing showed cancer chemopreventive properties at the early stages of diethylnitrosamine-induced hepatocarcinogenesis in rats. Possible inhibitory mechanisms include enhancement of the activities of some detoxifying enzymes and/or suppression of precancerous cells.

[Effects of filamentous macroalgae on the methane emission from urban river: a review].

The global warming caused by greenhouse gases emission has raised serious concerns. Recent studies found that the carbon dioxide (CO2) and methane (CH4) emissions from river ecosystem can partly offset the carbon sequestration by terrestrial ecosystem, leading to a rethink of the effects of river ecosystem on the global carbon balance and greenhouse gases emission inventory. As an important primary producer in urban river ecosystem, filamentous macroalgae can deeply affect the carbon cycle process of river system through changing the abiotic and biotic factors in the interface of water-sediment. This paper reviewed the effects of filamentous macroalgae on the CH4 emission from urban river system from the aspects of 1) the effects of urbanization on the river ecosystem and its CH4 emission flux, 2) the effects of filamentous macroalgae on the CH4 generation and emission process in natural river systems, and 3) the effects of filamentous macroalgae on the primary productivity and CH4 emission process in urban river systems. The current problems and future directions in related researches were discussed and prospected.

Characterization of gamma radiation inducible thioredoxin h from Spirogyra varians.

In this study, thioredoxin h (Trxh) was isolated and characterized from the fresh water green alga Spirogyra varians, which was one amongst the pool of proteins induced upon gamma radiation treatment. cDNA clones encoding S. varians thioredoxin h were isolated from a pre-constructed S. varians cDNA library. Trxh had a molecular mass of 13.5kDa and contained the canonical WCGPC active site. Recombinant Trxh showed the disulfide reduction activity, and exhibited insulin reduction activity. Also, Trxh had higher 5,5'-dithiobis(2-nitrobenzoic acid) reduction activity with Arabidopsis thioredoxin reductase (TR) than with Escherichia coli TR. Specific expression of the Trxh gene was further analyzed at mRNA and protein levels and was found to increase by gamma irradiation upto the absorbed dose of 3kGy, suggesting that Trxh may have potential functions in protection of biomolecules from gamma irradiation.

Study of the efficiency of immobilized algal technology for wastewater treatment.

The present paper deals with the study of efficiency of immobilized algal technology in wastewater treatment. The acclimatized algal species and wastewater samples were collected from three different sampling sites such as Kham River, Waluj [MIDC], Salim Ali Lake for the study. The encapsulation of collected algal species such as Spirogyra, Cyanobacteria in mixture form and Arthospira from selected sampling sites were made in sodium alginate for wastewater treatment. The percentage efficiency of immobilized algal technology for wastewater treatment was studied with respect to physico-chemical parameters. The physicochemical parameters were analyzed before and after treatment and compared for percentage efficiency study. The results obtained from present investigation reveal that the immobilized algal technology has maximum percentage efficiency in reduction of BOD, COD, Total Hardness, Total Alkalinity, Chloride. TSS, TDS and TSS. Whereas, the minimum fluctuations were found in pH and temperature. However, the immobilized algal technology is also useful to increase the dissolved oxygen concentration in wastewater treatment process.

Continuous metal removal from solution and industrial effluents using Spirogyra biomass-packed column reactor.

The granules of Spirogyra neglecta biomass, diameter 0.2-0.5mm, were successfully prepared by boiling it in urea-formaldehyde mixture. Metal sorption performance of the column packed with Spirogyra granules was assessed under variable operating conditions, such as, different influent metal concentrations, bed heights and flow rates. These conditions greatly influenced the breakthrough time and volume, saturation time and volume, and the ability of the column to attain saturation after reaching the breakthrough. The experimental breakthrough curves obtained under varying experimental conditions were modeled using Bohart-Adams, Wolborska, Thomas, Yoon-Nelson and modified dose-response models. The first two models were valid only in representing the initial part of the breakthrough curves; however, the other three models were good in representing the entire breakthrough curve. The granule-packed column could be successfully used up to 6 and 9 cycles of sorption and desorption for the removal of Cu(II) and Pb(II), respectively. The column could efficiently remove different metals from real industrial effluents, and hence the test biomass (Spirogyra granules) is a good candidate for commercial application.

The biosorption of heavy metals from aqueous solution by Spirogyra and Cladophora filamentous macroalgae.

The aim of this research was to develop a low cost adsorbent for wastewater treatment. The prime objective of this study was to search for suitable freshwater filamentous algae that have a high heavy metal ion removal capability. This study evaluated the biosorption capacity from aqueous solutions of the green algae species, Spirogyra and Cladophora, for lead (Pb(II)) and copper (Cu(II)). In comparing the analysis of the Langmuir and Freundlich isotherm models, the adsorption of Pb(II) and Cu(II) by these two types of biosorbents showed a better fit with the Langmuir isotherm model. In the adsorption of heavy metal ions by these two types of biosorbents, chemical and physical adsorption of particle surfaces was perhaps more significant than diffusion and adsorption between particles. Continuous adsorption-desorption experiments discovered that both types of biomass were excellent biosorbents with potential for further development.

Sorption properties of algae Spirogyra sp. and their use for determination of heavy metal ions concentrations in surface water.

Kinetics of heavy-metal ions sorption by alga Spirogyra sp. was evaluated experimentally in the laboratory, using both the static and the dynamic approach. The metal ions--Mn(2+), Cu(2+), Zn(2+) and Cd(2+)--were sorbed from aqueous solutions of their salts. The static experiments showed that the sorption equilibria were attained in 30 min, with 90-95% of metal ions sorbed in first 10 min of each process. The sorption equilibria were approximated with the Langmuir isotherm model. The algae sorbed each heavy metal ions proportionally to the amount of this metal ions in solution. The experiments confirmed that after 30 min of exposition to contaminated water, the concentration of heavy metal ions in the algae, which initially contained small amounts of these metal ions, increased proportionally to the concentration of metal ions in solution. The presented results can be used for elaboration of a method for classification of surface waters that complies with the legal regulations.