Non-invasive microelectrode cadmium flux measurements reveal the spatial characteristics and real-time kinetics of cadmium transport in hyperaccumulator and nonhyperaccumulator ecotypes of Sedum alfredii

This study aims to determine the spatial characteristics and real-time kinetics of cadmium transport in hyperaccumulator (HE) and non hyperaccumulator (NHE) ecotypes of Sedum alfredii using a non-invasive Cd-selective microelectrode. Compared with the NHE S. alfredii, the HE S. alfredii showed a higher Cd influx in the root apical region and root hair cells, as well as a significantly higher Cd efflux in the leaf petiole after root pre-treatment with cadmium chloride (CdCl₂). Thus, HE S. alfredii has a higher capability for the translocation of absorbed Cd to the shoot. Moreover, the mesophyll tissues, isolated mesophyll protoplasts, and intact vacuoles from HE S. alfredii exhibited an instantaneous influx of Cd in response to CdCl₂ treatment with mean rates that are markedly higher than those from NHE S. alfredii. Therefore, the hyper-accumulating trait of HE S. alfredii is characterized by the rapid Cd uptake in specific root regions, including the apical region and root hair cells, as well as by the rapid root-to-shoot translocation and the highly efficient Cd-permeable transport system in the plasma membrane and mesophyll cell tonoplast. We suggest that the non-invasive Cd-selective microelectrode is an excellent method with a high degree of spatial resolution for the study of Cd transport at the tissue, cellular, and sub-cellular levels in plants.     

Antiproliferative and Apoptosis Inducing Effects of Non-Polar Fractions from Lawsonia inermis L. in Cervical (HeLa) Cancer Cells

Two non-polar fractions viz. hexane (Hex-LI) and chloroform fraction (CHCl₃-LI) of Lawsonia inermis were studied for their antiproliferative potential in various cancer cell lines viz. HeLa, MCF-7, A549 and C6 glioma cells. Both the fractions showed more than 60 % of growth inhibition in all the tested cell lines at highest tested concentration. In clonogenic assay, different concentrations of Hex-LI and CHCl₃-LI decreased the number and size of colonies as compared to control in HeLa cells. The apoptotic effects as nuclear condensation, fragmentation were visualized with Hoechst-33342 staining of HeLa cells using confocal microscope. Both fractions induced apoptotic cell death in human cervical carcinoma (HeLa) cells as evident from flow cytometric analysis carried out using Annexin V-FITC and propidium iodide dyes. CHCl₃-LI treated cells significantly induced apoptosis (25.43 %) in comparison to control. Results from Neutral Comet assay demonstrated that both fractions induced double stranded breaks (DSB’s) in HeLa cells. Our data indicated that Hex-LI and CHCl₃-LI treated cells showed significant increase of 32.2 and 18.56 % reactive oxygen species (ROS) levels in DCFH-DA assay respectively. Further, experimental studies to decipher exact pathway via which these fractions induce cell death are in progress.     

Cadmium induces hypodermal periderm formation in the roots of the monocotyledonous medicinal plant Merwilla plumbea

Background and Aims

Merwilla plumbea is an important African medicinal plant. As the plants grow in soils contaminated with metals from mining activities, the danger of human intoxication exists. An experiment with plants exposed to cadmium (Cd) was performed to investigate the response of M. plumbea to this heavy metal, its uptake and translocation to plant organs and reaction of root tissues.

Methods

Plants grown from seeds were cultivated in controlled conditions. Hydroponic cultivation is not suitable for this species as roots do not tolerate aquatic conditions, and additional stress by Cd treatment results in total root growth inhibition and death. After cultivation in perlite the plants exposed to 1 and 5 mg Cd L⁻¹ in half-strength Hoagland''s solution were compared with control plants. Growth parameters were evaluated, Cd content was determined by inductively coupled plasma mass spectroscopy (ICP-MS) and root structure was investigated using various staining procedures, including the fluorescent stain Fluorol yellow 088 to detect suberin deposition in cell walls.

Key Results

The plants exposed to Cd were significantly reduced in growth. Most of the Cd taken up by plants after 4 weeks cultivation was retained in roots, and only a small amount was translocated to bulbs and leaves. In reaction to higher Cd concentrations, roots developed a hypodermal periderm close to the root tip. Cells produced by cork cambium impregnate their cell walls by suberin.

Conclusions

It is suggested that the hypodermal periderm is developed in young root parts in reaction to Cd toxicity to protect the root from radial uptake of Cd ions. Secondary meristems are usually not present in monocotyledonous species. Another interpretation explaining formation of protective suberized layers as a result of periclinal divisions of the hypodermis is discussed. This process may represent an as yet unknown defence reaction of roots when exposed to elemental stress.     

Azadirachta indica A. Juss (neem) induced morphological changes on oocytes of Rhipicephalus sanguineus (Latreille, 1806) (Acari: Ixodidae) tick females

Many studies have been conducted with plants whose extracts have the potential to be used for pest control. One of these plants is Azadirachta indica (neem), whose main active ingredient is azadirachtin, a compound shown to have acaricide and insecticide activity. Rhipicephalus sanguineus (brown dog tick) is currently considered to be an “urban pest,” because of its high levels of infestation and its ability to attack humans. In the present study partially and fully engorged R. sanguineus females were exposed to aqueous extracts of neem at concentrations of 10% and 20%, and to a control treatment. The results showed that differently from what was observed in the control, the pedicel cells of females exposed to neem at both concentrations lost their original shape. In the latter cases, the cytoplasm of the cells became fully vacuolated, especially near the germinal vesicle (oocyte nuclei) and in the oocyte pole, which is in contact with the cells of the pedicel. Oocytes in early stages of development (I and II) of ticks treated with both concentrations had irregular germinal vesicle, including the presence of two nucleoli as well as fragments of these. Oocytes in stages IV and V of exposed individuals showed full granular cytoplasm with bigger yolk granules when compared to the early stages. Chorion of mature oocytes was also altered, showing folds and deformations along their entire extension. The observed changes in cells of the reproductive system of R. sanguineus, especially in the oocytes, indicated the potential of neem as a new alternative method to control these ectoparasites.     

Oxidative damage and antioxidant responses in Microcystis aeruginosa exposed to the allelochemical berberine isolated from golden thread

Berberine, extracted from golden thread (Coptis chinensis Franch), is an allelochemical exhibiting inhibitory effects on the growth of Microcystis aeruginosa. Berberine-induced oxidative damage and antioxidant responses in M. aeruginosa cells were investigated to elucidate the mechanisms involved in berberine inhibition on algal growth. Malondialdehyde content in M. aeruginosa cells exposed to berberine increased with increased exposure concentration and the prolongation of exposure time. The same changes were observed in O₂⁻ activity of M. aeruginosa cells exposed to berberine. Berberine upregulated superoxide dismutase (SOD) activity at low concentrations while downregulating it at high concentrations. SOD activity transitioned from an increase to a decrease from 0 to 72 h exposure to 0.10% berberine. We observed that berberine exposure increased glutathione content in M. aeruginosa cells. The results suggested that berberine-induced oxidative damage might be at least partially responsible for berberine inhibition on M. aeruginosa growth.     

Analysis of inter-/intra-E-plate repeatability in the real-time cell analyzer

Over the past decade, the real-time cell analyzer (RTCA) has provided a good tool to the cell-based in vitro assay. Unlike the traditional systems that label the target cells with luminescence, fluorescence, or light absorption, RTCA monitors cell properties using noninvasive and label-free impedance measuring. However, realization of the maximum value of RTCA for applications will require assurance of within-experiment repeatability, day-to-day repeatability, and robustness to variations in conditions that might occur from different experiments. In this article, the performance and variability of RTCA is evaluated and a novel repeatability index (RI) is proposed to analyze the intra-/inter-E-plate repeatability of RTCA. The repeatability assay involves six cell lines and two media (water [H₂O] and dimethyl sulfoxide [DMSO]). First, six cell lines are exposed to the media individually, and time-dependent cellular response curves characterized as a cell index (CI) are recorded by RTCA. Then, the variations along sampling time and among repeated tests are calculated and RI values are obtained. Finally, a discriminating standard is set up to evaluate the degree of repeatability. As opposed to the standardized methodologies, it is shown that the presented index can give the quantitative evaluation for repeatability of RTCA within E-plate and variation on different days.     

Metallothionein and cellular energy allocation in the estuarine mysid shrimp Neomysis integer exposed to cadmium at different salinities

In the present study the induction of metallothioneins (MTs) and effects on the cellular energy allocation (CEA) in euryhaline crustacean Neomysis integer exposed to Cd at different salinities were studied. N. integer was exposed to the same sub-lethal concentration of free cadmium ion (1/5 of the cadmium activity of the reported 96 h LC₅₀ value) in hypo-osmotic (7.2 μg Cd/L at 5 psu), isosmotic (23.0 μg Cd/L at 16 psu) and hyper-osmotic media (38.1 μg Cd/L at 25 psu) for 7 days. By using the free Cd concentration as the basis for conducting the exposures, the effect of salinity on cadmium speciation was eliminated and therefore the true effect of salinity as an abiotic factor on the MT induction and CEA could be studied.MT content was quantified by differential pulse voltammetry (DPV); this is the first time that this method is applied to assays with N. integer. No significant differences in MT levels between the control and Cd-exposed groups were observed. The measured MT levels (ranging from 32.3 to 75.7 μg/mg_{(cytosolic protein)}) probably represent the constitutive MT levels responsible for binding essential metals. The differences in MT levels observed at the different salinities indicate a possible relationship between some physiological process (possibly osmoregulation) other than detoxification and MT induction. A decrease in salinity caused an increase in MT level in N. integer, and this was significantly correlated to the CEA values (Spearman correlation coefficient r = − 0.56, p = 0.016). Our results indicate that salinity can change the energy status and MT content of N. integer. These findings should be taken into account when using these biomarkers - and especially MTs - in field studies and environmental monitoring.     

Micronucleus frequency and lipid peroxidation in <Emphasis Type="Italic">Allium sativum</Emphasis> root tip cells treated with gibberellic acid and cadmium

Gibberellic acid (GA₃) is a very potent hormone whose natural occurrence in plants controls their development. Cadmium is a particularly dangerous pollutant due to its high toxicity and great solubility in water. In this study, the effect of GA₃ on Allium sativum root tip cells was investigated in the presence of cadmium. A. sativum root tip cells were exposed to CdNO₃ (50, 100, 200 μM), GA₃ (10-3 M), both CdNO₃ and GA₃. Cytogenetic analyses were performed as micronucleus (MN) assay and mitotic index (MI). Lipid peroxidation analysis was also performed in A. sativum root tip cells for determination of membrane damage. MN exhibited a dose-dependent increase in Cd treatments in A. sativum. GA₃ significantly reduced the effect of Cd on the MN frequency. MN was observed in GA₃ and GA₃ + 50 μm Cd treatments at very low frequency. MI slightly decreased in GA₃ and GA₃ + Cd treatments. MI decreased more in high concentrations of Cd than combined GA₃ + Cd treatments. The high concentrations of cadmium induce MN, lipid peroxidation and lead to genotoxicity in A. sativum. Current work reveals that the effect of Cd on genotoxicity can be partially restored with GA₃ application.     

Characterization of abalone Haliotis tuberculata–Vibrio harveyi interactions in gill primary cultures

The decline of European abalone Haliotis tuberculata populations has been associated with various pathogens including bacteria of the genus Vibrio. Following the summer mortality outbreaks reported in France between 1998 and 2000, Vibrio harveyi strains were isolated from moribund abalones, allowing in vivo and in vitro studies on the interactions between abalone H. tuberculata and V. harveyi. This work reports the development of primary cell cultures from abalone gill tissue, a target tissue for bacterial colonisation, and their use for in vitro study of host cell—V. harveyi interactions. Gill cells originated from four-day-old explant primary cultures were successfully sub-cultured in multi-well plates and maintained in vitro for up to 24 days. Cytological parameters, cell morphology and viability were monitored over time using flow cytometry analysis and semi-quantitative assay (XTT). Then, gill cell cultures were used to investigate in vitro the interactions with V. harveyi. The effects of two bacterial strains were evaluated on gill cells: a pathogenic bacterial strain ORM4 which is responsible for abalone mortalities and LMG7890 which is a non-pathogenic strain. Cellular responses of gill cells exposed to increasing concentrations of bacteria were evaluated by measuring mitochondrial activity (XTT assay) and phenoloxidase activity, an enzyme which is strongly involved in immune response. The ability of gill cells to phagocyte GFP-tagged V. harveyi was evaluated by flow cytometry and gill cells-V. harveyi interactions were characterized using fluorescence microscopy and transmission electron microscopy. During phagocytosis process we evidenced that V. harveyi bacteria induced significant changes in gill cells metabolism and immune response. Together, the results showed that primary cell cultures from abalone gills are suitable for in vitro study of host-pathogen interactions, providing complementary assays to in vivo experiments.     

Poly(gamma-glutamylcysteinyl)glycine Synthesis in Datura innoxia and Binding with Cadmium : Role in Cadmium Tolerance

The effects of Cd on poly(γ-glutamylcysteinyl)glycine [(γEC)_nG] biosynthesis and formation of (γEC)_nG:Cd complexes were measured in two cell lines of Datura innoxia with differing Cd tolerance. In addition, RNA synthesis, protein synthesis, and GSH concentrations were measured during a 48 hour exposure to Cd. Exposure to 250 micromolar CdCl₂ was toxic to the sensitive line, whereas the tolerant line survived and grew in its presence. Cd-sensitive cells synthesized the same amount of (γEC)_nG as tolerant cells during an initial 24 hour exposure to 250 micromolar CdCl₂. However, rates of (γEC)_nG:Cd complex formation differed between the two cell lines with the sensitive cells forming complexes later than tolerant cells. In addition, the complexes formed by sensitive cells were of lower molecular weight than those of tolerant cells and did not bind all of the cellular Cd. Pulse-labeling of cells with l-[³⁵S]cysteine resulted in equivalent rates of incorporation into the (γEC)_nG of both cell lines during the initial 24 hours after Cd. Rates of protein and RNA synthesis were similar for both cell lines during the initial 8 hours after Cd but thereafter declined rapidly in sensitive cells. This was reflected by a decline in viability of sensitive cells. The GSH content of both cell lines declined rapidly upon exposure to Cd but was higher in sensitive cells throughout the experiment. These results show that the biosynthetic pathway for (γEC)_nG synthesis in sensitive cells is operational and that relative overproduction of (γEC)_nG is not the mechanism of Cd-tolerance in a Cd-tolerant cell line of D. innoxia. Rapid formation of (γEC)_nG:Cd complexes that bind all of the cellular Cd within 24 hours appears to correlate with tolerance in these cells.     

可降解螯合剂对镉胁迫下籽粒苋根系形态及生理生化特征的影响

采用盆栽试验研究了可降解螯合剂EDDS和NTA对镉胁迫下籽粒苋(Amaranthus hybridus L.)根系形态及生理生化特征的影响。结果表明:当螯合剂施入10 mg/kg的镉污染土壤后,籽粒苋根系生物量和总长等根系形态指标与对照无显著差异,过氧化物酶(POD)、过氧化氢酶(CAT)活性、谷胱甘肽(GSH)和可溶性蛋白含量显著上升。当螯合剂施入100 mg/kg的镉污染土壤后,籽粒苋根系生物量、总长、表面积、体积及侧根数比对照显著减少了12.30%—23.98%、17.01%—24.90%、41.87%—57.93%、16.46%—32.94%和23.48%—53.35%;EDDS的施入使籽粒苋根系POD、CAT活性、GSH和可溶性蛋白含量显著升高;而NTA施入后,根系中的POD活性比对照降低了4.12%—35.95%,并且CAT活性和可溶性蛋白含量在2 mmol/kg NTA处理下分别显著降低了14.66%—15.79%和26.81%—30.48%;EDDS和NTA施入后,GSH含量比对照显著升高了14.73%—65.65%和28.05%—84.10%。当镉处理浓度分别为10 mg/kg和100 mg/kg时,螯合剂的施入显著增强了籽粒苋根系对镉的吸收,比对照分别增加了40.76%—103.10%和15.03%—49.49%。因此,EDDS和NTA施入镉污染土壤后,通过影响籽粒苋根系形态和生理生化过程以响应重金属镉的胁迫。     

Genista sessilifolia DC. and Genista tinctoria L. inhibit UV light and nitric oxide-induced DNA damage and human melanoma cell growth

Many Genista species (Leguminosae), containing isoflavones as biologically active substances, show interesting biological properties such as hypoglycemic, antiinflammatory, antiulcer, spasmolytic, antioxidant, estrogenic and cytotoxic activity against different human cancer cell lines. In this work, we describe the chemical composition of the methanolic extracts from aerial parts of Genista sessilifolia DC. and Genista tinctoria L., and their biological activity testing the effect on pBR322 DNA cleavage induced by hydroxyl radicals (OH), generated from UV-photolysis of hydrogen peroxide (H₂O₂) and by nitric oxide (NO). In addition, we investigated the growth inhibitory activity of these natural products against human melanoma cell line (M14). The extracts of G. sessilifolia and G. tinctoria, for their isoflavone components, showed a protective effect on UV light and nitric oxide-mediated plasmid DNA damage, and inhibited the growth of melanoma cells. The data of the present study also suggest that these natural products could trigger apoptotic death in M14 cells. In fact, a high DNA fragmentation (COMET assay) and a significant increase of caspase-3 activity, not correlated to LDH release, a marker of membrane breakdown, occurred in melanoma cells exposed to these extracts. The significant production of reactive oxygen species (ROS) evidenced in these experimental conditions could contribute to trigger the apoptosis cascades.     

Cathepsin B-like and cell death in the unicellular human pathogen Leishmania

In several studies reporting cell death (CD) in lower eukaryotes and in the human protozoan parasite Leishmania, proteolytic activity was revealed using pan-caspase substrates or inhibitors such as carbobenzoxy-valyl-alanyl-aspartyl-[O-methyl]-fluoromethylketone (Z-VAD-FMK). However, most of the lower eukaryotes do not encode caspase(s) but MCA, which differs from caspase(s) in its substrate specificity and cannot be accountable for the recognition of Z-VAD-FMK. In the present study, we were interested in identifying which enzyme was capturing the Z-VAD substrate. We show that heat shock (HS) induces Leishmania CD and leads to the intracellular binding of Z-VAD-FMK. We excluded binding and inhibition of Z-VAD-FMK to Leishmania major metacaspase (LmjMCA), and identified cysteine proteinase C (LmjCPC), a cathepsin B-like (CPC) enzyme, as the Z-VAD-FMK binding enzyme. We confirmed the specific interaction of Z-VAD-FMK with CPC by showing that Z-VAD binding is absent in a Leishmania mexicana strain in which the cpc gene was deleted. We also show that parasites exposed to various stress conditions release CPC into a soluble fraction. Finally, we confirmed the role of CPC in Leishmania CD by showing that, when exposed to the oxidizing agent hydrogen peroxide (H₂O₂), cpc knockout parasites survived better than wild-type parasites (WT). In conclusion, this study identified CPC as the substrate of Z-VAD-FMK in Leishmania and as a potential additional executioner protease in the CD cascade of Leishmania and possibly in other lower eukaryotes.     

Cadmium stress stimulates tissue turnover in Helix pomatia: increasing cell proliferation from metal tolerance to exhaustion in molluscan midgut gland

In terrestrial pulmonate snails, cadmium (Cd) uptake leads to the induction of a Cd-specific metallothionein isoform (Cd-MT) that protects against adverse interactions of this toxic metal ion. Increasing concentrations of Cd cause increased individual mortality possibly linked to pathological alterations in the snail midgut gland. Histological, immuno-histochemical, and electron-microscopic methods in combination with tissue metal analyses and quantification of MT induction parameters were applied to the midgut gland of Cd-exposed Roman snails (Helix pomatia). Conspicuous concentration-dependent alterations occurred in this organ, including the metal-induced increase of Cd-MT concentration and manifestation of Cd-MT mRNA precipitations in all midgut gland cell types. The most evident alteration was an increase of cellular turnover reflected by enhanced cell proliferation. Intensified vesiculation of endoplasmic reticulum was noted in basophilic cells and an increasing formation of lipofuscin granules in excretory cells. At the highest Cd concentrations, mitochondrial membranes were disrupted in basophilic cells, and lipofuscin granules were released from excretory cells into the midgut gland tubular system. Some of these alterations (e.g., increased cell proliferation rate, vesiculation of endoplasmic reticulum) detected at low Cd concentrations were interpreted as adaptive response processes enhancing the tolerance of exposed individuals to metal stress. Cellular alterations at higher Cd concentrations (e.g., mitochondrial structural damage) clearly represented ongoing irreversible cellular disruption. Combined evaluation of cellular biomarkers and MT saturation levels indicated that the transition from stress resistance to depletion of resistance capacity occurred above a threshold of 0.8 µmol Cd/g dry weight in the midgut gland of H. pomatia. At these Cd concentrations, Cd-MT was saturated with Cd²⁺ ions, whereas at the cellular level, structural alterations turned into pathological deterioration.     

Cadmium(II)-stimulated enzyme activation of Arabidopsis thaliana phytochelatin synthase 1

Phytochelatin (PC), a class of heavy metal-binding peptides, is synthesized from the tripeptide glutathione (GSH) and/or previously synthesized PC in a reaction mediated by PC synthase (PCS). In the present study, the PC production rate catalyzed by recombinant Arabidopsis PCS1 (rAtPCS1) in the presence of a constant free Cd(II) level increased steadily and the kinetic parameters were approximated using a substituted-enzyme mechanism in which GSH and bis(glutathionato)cadmium acted as co-substrates. In contrast, the PC production rate as a function of GSH concentration at a constant total Cd(II) concentration reached a maximum, which shifted toward higher GSH concentrations as the concentration of Cd(II) was increased. These observations are consistent with the suggestion that rAtPCS1 possesses a Cd(II) binding site where Cd(II) binds to activate the enzyme. The affinity constant, optimized using a one-site mathematical model, successfully simulated the experimental data for the assay system using lower concentrations of Cd(II) (5 or 10 μM) but not for the assay using higher concentrations (50 or 500 μM), where a sigmoidal increase in PCS activity was evident. Furthermore, the PCS activity determined at a constant GSH concentration as a function of Cd(II) concentration also reached a maximum. These findings demonstrate that rAtPCS1 also possesses a second Cd(II) binding site where Cd(II) binds to induce an inhibitory effect. A two-site mathematical model was applied successfully to account for the observed phenomena, supporting the suggestion that rAtPCS1 possesses two Cd(II) binding sites.     

Sensitivity of Meloidogyne incognita and Rotylenchulus reniformis to Abamectin

Avermectins are macrocyclic lactones produced by Streptomyces avermitilis. Abamectin is a blend of B_1a and B_1b avermectins that is being used as a seed treatment to control plant-parasitic nematodes on cotton and some vegetable crops. No LD₅₀ values, data on nematode recovery following brief exposure, or effects of sublethal concentrations on infectivity of the plant-parasitic nematodes Meloidogyne incognita or Rotylenchulus reniformis are available. Using an assay of nematode mobility, LD₅₀ values of 1.56 μg/ml and 32.9 μg/ml were calculated based on 2 hr exposure for M. incognita and R. reniformis, respectively. There was no recovery of either nematode after exposure for 1 hr. Mortality of M. incognita continued to increase following a 1 hr exposure, whereas R. reniformis mortality remained unchanged at 24 hr after the nematodes were removed from the abamectin solution. Sublethal concentrations of 1.56 to 0.39 μg/ml for M. incognita and 32.9 to 8.2 μg/ml for R. reniformis reduced infectivity of each nematode on tomato roots. The toxicity of abamectin to these nematodes was comparable to that of aldicarb.     

Cadmium accumulation in Atriplex halimus subsp. schweinfurthii and its influence on growth,proline, root hydraulic conductivity and nutrient uptake

Atriplex halimus subsp. schweinfurthii is a newly found cadmium (Cd)-hyperaccumulator, but there have been no detailed studies on its physiological responses when Cd is hyperaccumulated. A. halimus was grown in hydroponic conditions to investigate the effect of cadmium chloride (CdCl₂) on growth, water status, leaf chlorophyll concentration, proline and Cd accumulation. Treatments were prepared by adding 0, 50, 100, 200 and 400 μM CdCl₂ to the nutrient medium. Plant growth was significantly affected at high-Cd treatments. Increased CdCl₂ decreased chlorophyll concentration, transpiration and root hydraulic conductivity (L₀). Hence water flux had only a little effect on the uptake of Cd in A. halimus seedlings. In contrast, proline content increased with increasing CdCl₂ concentration. Plants accumulated substantial amount of Cd in different plant parts (shoot and root). Most of the Cd taken up was retained in roots (606.51 μg g⁻¹DW after 15 d at 400 μM CdCl₂). The addition of Cd in the culture medium affected calcium (Ca) and potassium (K) nutrition in both shoot and root. A. halimus provides a new plant resource for exploring the mechanism of Cd hyperaccumulation and has potential for use in the phytostabilization of Cd-contaminated salt soils.     

In-Vitro and in-Silico characterization of Sophora interrupta plant extract as an anticancer activity

Sophora interrupta belongs to the family of Fabaceae and the species in this genus have a diverse medicinal importance as a folk medicine for preventing many ailments including cancer. In order to evaluate the anticancer activity of S.interrupta, we have performed in vitro anti-oxidant, anti-inflammatory, anti-proliferative, and cell based anticancer activity in MCF-7 and PC-3 cell lines. Secondary metabolites of S.interrupta were used to identify anticancer compounds using Open Eye software. The antioxidant activity of the S.interrupta root ethylacetate (SEA) extract at 100 µg/ml is equal to that of ascorbic acid at 50 µg/ml. The antiinflammatory activity of SEA is half of that of diclofenac at 50 µg/ml. Anticancer activity was detected by measuring the mitochondrial dehydrogenase activity (MTT assay). The half maximal inhibitory concentrations (IC₅₀) for MCF-7 and PC-3 cell lines are 250 and 700 µg/ml respectively. This was supported by the morphological changes such as membrane blebbing, cell detachment and rounded cell morphology when compared to the parental cells. In addition, we observed few green cells (live) over red cells (dead) based on the uptake of acridine orange and ethidium bromide dyes. Kaempferol-3-O-b-D-glucopyranoside, a Secondary metabolite of S.interrupta form 6 hydrogen bond interactions with Arg 202, Gln 207, Gly 227, Gly 229, Thr 231 and Ala 232 human DEAD box RNA helicase, DDX3 protein and is equivalent to crystal structure of adenosine mono phosphate to DDX3. Overall, it suggests that the SEA extract has anticancer compounds, and it can be used to enhance death receptor mediated cancer cell death.     

Characteristics of Cd uptake, translocation and accumulation in a novel Cd-accumulating tree, star fruit (Averrhoa carambola L., Oxalidaceae)

Cadmium (Cd) accumulation by terrestrial higher plants is an intriguing phenomenon that may be exploited for phytoextraction of Cd-contaminated soils. Characterizing the physiological processes responsible for elevated concentrations of Cd in shoots is a first step towards a comprehensive understanding of the mechanisms underlying Cd accumulation in plants and may eventually improve the efficiency of phytoextraction. Woody species that can accumulate Cd have been recently recommended as good candidates for phytoextraction of Cd-contaminated soils. However, little is known about the mechanisms of Cd accumulation by woody species. In an attempt to understand the physiological processes contributing to Cd accumulation in woody species, Cd uptake and translocation by a novel tropical Cd-accumulating tree, star fruit (Averrhoa carambola) were characterized and compared with those of a non-Cd-accumulating tree (Clausena lansium). Our results showed that A. carambola had higher Cd uptake and root-to-shoot translocation efficiencies than C. lansium, which might account for its greater Cd-accumulating capacity. Furthermore, Cd accumulation by A. carambola was not significantly affected by zinc (Zn), whereas Zn accumulation was greatly lowered by Cd. This phenomenon could not be fully explained by a simple competition between Cd²⁺ and Zn²⁺, implying the existence of a transport system with a preference for Cd over Zn. Collectively, our results indicate that A. carambola has noteworthy physiological traits associated with accumulation of Cd to high levels.