NaCl 150mM Research Articles

Pretreatment with LEDs regulates antioxidant capacity and polyphenolic profile in two genotypes of basil under salinity stress.

In the present study, we evaluated a pretreatment with four LED light sources (red, blue, red + blue, and white) in two genotypes (green and purple) of basil on the growth parameters, stress oxidative markers, non-enzymatic antioxidants, osmoprotectant compounds, ion content, and polyphenolic profile under both control and salinity stress conditions. The results indicated that 150mM of NaCl decreased biomass, RWC, and K+/Na+ ratio but increased the content of proline and antioxidant capacity in the leaves of both genotypes of basil grown under GH (greenhouse) conditions. The results suggested that RB LED-exposed plants in the green genotype and R LED-exposed plants in the purple genotype improved accumulation of shoot biomass, K+/Na+ ratio, proline and soluble sugars, glutathione and ascorbate, polyphenolic profile, and thioredoxin reductase activity in the leaves of basil under both control and salinity stress conditions. NaCl stress (150mM) increased oxidative markers, which are responsible for disturbance of routine functions of various plantcellular modules. LED light pretreatments diminished these markers under both control and salinity stress conditions. It could be concluded that intensification of non-enzymatic antioxidant systems during light-mediated priming can diminish the deleterious effects of ROS induced by NaCl stress (150mM) through preventing the lipid peroxidation, scavenging cytotoxic H2O2, and enhancement of antioxidant potentials. Therefore, usage of LED lighting systems as a pretreatment or to supplement natural photoperiods under both control and salinity stress conditions may be advantageous for increasing biomass and phytochemical accumulation in basil.

Salinity decreases cadmium accumulation in Vicia faba.

The study investigates the effect of cadmium (Cd), salinity (NaCl), and combined stress on rhizosphere pH, growth parameters, membrane leakage, and genotoxicity in Vicia faba. Germinated seeds were exposed for 48h to 0.01mM Cd(NO3)2 (Cd), 50mM NaCl (S50), 150mM NaCl (S150), and Cd-NaCl (CdS50 and CdS150). An accumulation of Cd and Na was found essentially in Vicia roots under each single stress factor associated with variations in rhizosphere pH. Additional NaCl in metallic solution significantly dropped the rhizosphere pH and decreased Cd concentrations in roots by 2.3 and 3.8 times for CdS50 and CdS150, respectively. Growth parameters (root length and fresh and dry matters), mitotic activity, and micronucleus formation were not influenced by Cd and low concentration of NaCl when applied separately or together, while 150mM of NaCl, alone or combined with Cd, affected negatively all the studied parameters, as well as chromosome and nucleus stability. V. faba seems to reduce the transport of Cd in saline conditions and therefore salinity (50mM) may act as a protection against Cd accumulation.

Priming Induction in Neighbouring Plants of Gossypium hirsutum under Salt Stress

Plants are subjected to various types of environmental stresses throughout their lifecycle. It has been found that plants are able to communicate with the neighbouring plants under stress conditions through volatile organic compounds. These volatiles act as signals for the neighbouring plants thus preparing them for the upcoming stress, a phenomenon known as priming. So, the present study explores the effects of salt stress on cotton plants and the resultant induction of priming in the nearby plants. For this purpose, salt tolerant cotton (Gossypium hirsutum) variety was used. Two concentration levels, 100 mM, and 150 mM of salt were used to study the impacts of the stress. The experiment was divided into two steps for each treatment. In the first step, a set of plants (emitters) was given salt stress. The second set of plants (receivers) was placed adjacent to the stressed plants (emitters), while the third set of plants was placed separately as a control for both the treatments. Various physiological and morphological parameters were measured at the beginning and the end of the first step. In the second step, the receiver plants now termed as "primed" were given the same levels of stress while a new set of non-primed plants was placed near the primed plants. These non-primed plants were now treated with 100mM and 150mM of NaCl respectively and the results were compared. The results show that plants were able to get signals from neighbouring stressed plants. Plants responded by altering morphology and physiology to prepare themselves for future stress conditions.

Biochemical performance and protein profile of sensitive and tolerant varieties of chickpea under salinity

Chickpea varieties (BG-256 and CSG-8962) were subjected to salt stress (50mM, 100mM and 150mM of NaCl) in a pot experiment. Untreated plants served as control. Plants were analyzed from 20 DAS up to 60 DAS at ten-day interval. Salt stress significantly reduced growth parameters like biomass, net assimilation rate, relative water content and biochemical parameters viz., total nitrogen and protein content of both the varieties. However, decrement was more pronounced in sensitive (BG-256) than in tolerant (CSG-8962) variety. Proline content increased with increase in salt exposure in both the varieties. SDS-PAGE of the protein reveal large amount of protein degradation in plants treated with high concentration of salt.

Structural and functional characterisation of two novel durum wheat annexin genes in response to abiotic stress.

Abiotic stress results in massive loss of crop productivity throughout the world. Understanding the plant gene regulatory mechanisms involved in stress responses is very important. Annexins are a conserved multigene family of Ca-dependent, phospholipid-binding proteins with suggested functions in response to environmental stresses and signalling during plant growth and development. Annexins function to counteract oxidative stress, maintain cell redox homeostasis and enhance drought tolerance. A full-length cDNA of two genes (TdAnn6 and TdAnn12) encoding annexin proteins were isolated and characterised from Tunisian durum wheat varieties (Triticum turgidum L. subsp. durum cv. Mahmoudi). Analyses of the deduced proteins encoded by annexin cDNAs (TdAnn6 and TdAnn12) indicate the presence of the characteristic four repeats of 70-75 amino acids and the motifs proposed to be involved in Ca2+ binding. Gene expression patterns obtained by real-time PCR revealed differential temporal and spatial regulation of the two annexin genes in durum wheat under different abiotic stress conditions such as salt (NaCl 150mM), osmotic (10% polyethylene glycol 8000), ionic (LiCl 10mM), oxidative (H2O2), ABA (100µM), salicylic acid (10mM), cold (4°C) and heat (37°C) stress. The two annexin genes were not regulated by heavy metal stress (CdCl2 150µM). Moreover, heterologous expression of TdAnn6 and TdAnn12 in yeast improves its tolerance to abiotic stresses, suggesting annexin's involvement in theses stress tolerance mechanisms. Taken together, our results show that the two newly isolated wheat annexin might play an active role in modulating plant cell responses to abiotic stress responses.

Interactive effect of salinity and zinc stress on growth and photosynthetic responses of the perennial grass, Polypogon monspeliensis

A glasshouse experiment was designed to investigate the interactive effect of zinc from 0 to 300ppm and/or 150mM NaCl on the growth and the photosynthetic apparatus of Polypogon monspeliensis from two Tunisian provenances (North and South) seedlings by measuring Growth, photosynthesis, chlorophyll fluorescence and chlorophyll concentration after 30days of treatments. These parameters were severely reduced by this heavy metal. The zinc excess involves the degradation of photosystem II and the chlorophyll decomposition. However, these effect of Zn concentrations alleviated by 150mM of NaCl. Adequate Zn also prevents uptake and accumulation of Na and Cl− in shoot, by increasing membrane integrity of root cells. Our results show that at the same time, the variation of these parameters depends on the seeds origin, Zn stress and saline conditions.

Highly stable polyglutamate derivatives/siRNA polyplex efficiently down-relegate survivin expression and augment the efficacy of cisplatin

RNA interfere (RNAi)-based technology holds great promise in cancer treatment. The use of small interfering RNA (siRNA), however, is hampered by its low delivery efficiency in vivo when they are diluted in blood biofluids and in the presence of serum and salt. In this study, we developed the polyglutamate derivative polymer brush, poly(ethyleneglycol) monomethyl ether-b-polyglutamate-g-spermine (mPEG-b-PG-g-spermine, PPGS), which could efficiently deliver survivin-siRNA under ultra-high dilution and in the presence of salt (NaCl 150mM) and serum (10% FBS), most likely due to its PEG-shelled polymer brush structure. On the contrary, aggregation occurred when PEI/siRNA polyplex dispersed in saline and serum-containing media and PEI polyplex dissociated after making a 256-fold dilution. PPGS/si-survivin polyplex exhibited high cellular uptake efficiency and efficiently down-regulated the expression of survivin mRNA in the cisplatin-resistance of non-small cell human lung adenocarcinoma (A549/DDP) cells in the presence of serum. However, either PEI polyplex or Lipofectmine 2000 complex was unstable in serum and salt-containing media and at high dilution rates, which resulted in their dramatical decrease of cellular uptake and gene-silencing efficiency in these conditions. The PPGS/si-survivin polyplex also exhibited synergistic effects of killing the cancer cells by combination treatment with cisplatin. Therefore, the PPGS gene carrier showed great potential in systemic siRNA delivery, and its combination with chemotherapeutic drug is promising in treating drug resistant cancers.

Screening Six Varieties of Rice (Oryzasativa) for Salinity Tolerance

Salinity is a constraint to the sustainable agricultural production. Increasing salinity is found to inhibit growth, height, and total of leaf area ofrice from vegetative to generative stages. Electrophysiologically, increasing salinity and exposure duration reduce the PD of the plant leaves. The screening focuses on the response of six varieties of rice (Oryza sativa) grown in a glasshouse subjected to five different salinity levels [0mM; 50mM; 100mM; 150mM and 200mM of NaCl]. Death occurred at 200mM of NaCl though all six varieties were tolerant to 100mMofNaClduring 4 weeks of application, and four varieties of rice were found to be sensitive to salinity around 150mM of NaCl during 3 weeks of application.In turn, this study will help increase the security of the major food demand consumed by around 90% of Indonesian and protect the livelihood sustainability of around 67% Indonesian farmers, and help reduce starvation risk of a community due to salinity effects.

Tolerance of Miscanthus × giganteus to salinity depends on initial weight of rhizomes as well as high accumulation of potassium and proline in leaves

Miscanthus×giganteus, a species useful for sustainable agriculture, can be cultivated on soils with lower bonitation and also with higher salinity. The study aimed to estimate NaCl concentrations which limit growth of Miscanthus×giganteus plants and to determine physiological parameters best reflecting the response of Miscanthus to salt stress. The experiment was conducted on plants grown at 0–200mM of NaCl. All analyses were done after 12 weeks of salt treatment. In rhizomes and leaves, fresh (FW) and dry weight (DW), relative water content (RWC), electrolyte leakage (EL), and ion content were determined; in leaves only, chlorophyll fluorescence, net photosynthesis rate (PN), transpiration (E), stomatal conductance (gs) and proline content. Shoots developed at all NaCl concentrations, but at 200mM many rhizomes produced none; thus analyses on leaves were done only on plants grown at 0–150mM of NaCl. Salinity decreased FW and DW of roots and shoots significantly, and retarded increases in rhizome weight. In leaves, salinity of 100 and 150mM NaCl did not damage membrane structures, while in rhizomes only 200mM increased EL considerably. Most parameters of chlorophyll fluorescence and gas exchange declined under salinity. High amounts of proline, K and Na were accumulated in leaves at 100 and 150mM NaCl. RWC remained unchanged in roots and leaves and decreased in rhizomes under rising salinity. The initial FW of the rhizomes studied correlated with the salt tolerance of Miscanthus plants. Changes in chlorophyll fluorescence, PN, FW and DW of roots and leaves as well as potassium and proline concentrations in leaves are the most reliable indicators of the response of Miscanthus to salinity. A high accumulation of K+ and proline in leaves reduces the sodium effect and determines the tolerance of Miscanthus to salinity.

Growth stage-based modulation in antioxidant defense system and proline accumulation in two hexaploid wheat (Triticum aestivum L.) cultivars differing in salinity tolerance

Changes in the antioxidant defense system and proline accumulation were examined at different growth stages (vegetative, boot and reproductive) in plants of two hexaploid spring wheat cultivars (S-24, salt tolerant; MH-97, salt sensitive), grown in hydroponics and salinity-affected with 0, 50, 100 and 150mM of NaCl. Salt stress provoked a marked decline in plant dry mass and ascorbic acid contents, and increased proline, total soluble proteins and H2O2 contents in both wheat cultivars at all growth stages. However, higher accumulation of proline and H2O2 contents was recorded at the vegetative and boot stages, respectively, in both wheat cultivars. Salt stress caused a consistent rise in the activities of some key antioxidant enzymes (CAT, SOD, POD, and APX) at all growth stages only in the salt tolerant cultivar S-24, whereas such pattern of enhanced activities of enzymatic antioxidants in cv. MH-97 was found only at the vegetative stage under saline regimes. Maximum activities of various enzymatic antioxidants were observed at the vegetative stage in both wheat cultivars under varying external salt treatments. The results showed that high salinity tolerance of cv. S-24, as manifested by lower decrease in its dry matter under salt stress, was associated with higher activities of antioxidant enzymes, increased accumulation of proline, and lower levels of H2O2, as compared with cv. MH-97 at all growth stages under saline regimes.

Alleviation of salt stress in lemongrass by salicylic acid

Soil salinity is one of the key factors adversely affecting the growth, yield, and quality of crops. A pot study was conducted to find out whether exogenous application of salicylic acid could ameliorate the adverse effect of salinity in lemongrass (Cymbopogon flexuosus Steud. Wats.). Two Cymbopogon varieties, Krishna and Neema, were used in the study. Three salinity levels, viz, 50, 100, and 150mM of NaCl, were applied to 30-day-old plants. Salicylic acid (SA) was applied as foliar spray at 10(-5)M concentration. Totally, six SA-sprays were carried out at 10-day intervals, following the first spray at 30days after sowing. The growth parameters were progressively reduced with the increase in salinity level; however, growth inhibition was significantly reduced by the foliar application of SA. With the increase in salt stress, a gradual decrease in the activities of carbonic anhydrase and nitrate reductase was observed in both the varieties. SA-treatment not only ameliorated the adverse effects of NaCl but also showed a significant improvement in the activities of these enzymes compared with the untreated stressed-plants. The plants supplemented with NaCl exhibited a significant increase in electrolyte leakage, proline content, and phosphoenol pyruvate carboxylase activity. Content and yield of essential oil was also significantly decreased in plants that received salinity levels; however, SA overcame the unfavorable effects of salinity stress to a considerable extent. Lemongrass variety Krishna was found to be more adapted to salt stress than Neema, as indicated by the overall performance of the two varieties under salt conditions.

Reversal of Glanzmann's Thrombasthenia & Glycoprotein IIb/Iiia Antagonists by Combined Haemostatic Intervention with Fibrinogen Concentrate and Recombinant Factor VIIa.

Abstract 1325Poster Board I-347 IntroductionGlanzmann's thrombasthenia (GT) is a rare platelet dysfunction caused by deficiency of gp IIb/IIIa. Treatment with platelet glycoprotein (gp) IIb/IIIa antagonists is common practice in patients undergoing urgent percutaneous cardiac intervention and causes a phenotypic defect similar to GT. Urgent need of acute open-heart surgery may cause excessive bleeding due to the presence of gpIIb/IIIa antagonists and access to platelet transfusion may cause delay. AimThe present laboratory study explores treatment modalities for reversal of GT and platelet gpIIb/IIIa antagonists, based on recombinant factor VIIa (rFVIIa), fibrinogen, as well as combination of the two drugs. Materials and methodsA patient with severe GT, and healthy volunteers provided whole blood (WB) samples for the study. Blood samples were taken in tubes containing 3.2% citrate and corn trypsin inhibitor (final conc. 18.3 μg/mL) to abolish artificial contact activation. Blood samples from healthy volunteers were spiked with aliquots of buffer (HEPES 20mM, NaCl 150mM, pH=7.4) and abciximab reaching final concentrations of 1, 2, 4, 8, 16, 32, and 64 μg/ml. Subsequently, blood samples treated with abciximab 32 μg/ml were spiked with fibrinogen concentrate (0.5, 1, 2, 4, 8 mg/mL) or rFVIIa (1, 2, 4, 8 μg/mL) as well as the combination of fibrinogen and rFVIIa. Haemostatic capacity was evaluated by recording dynamic WB ROTEM thromboelastometry using activation with minute amounts of tissue factor (TF, Innovin̈, final dilution 1:17,000) as well as clot stability using TF + tissue plasminogen activator (t-PA, final concentration 2 nM). WB platelet adhesion was visualized by Impact-R̈. Data was visualized graphically and interpreted using descriptive statistics. ResultsTitration experiments with abciximab increased the clotting time (CT) and reduced the maximum clot firmness (MCF) (Figure 1). The patients clotting profiles were comparable to the highest dose of abciximab. In vitro addition of fibrinogen concentrate dose dependently reversed the MCF, but further prolonged the CT (Figure 2). In contrast, rFVIIa significantly shortened the CT, whereas is had limited effect on the MCF (Figure 2). Combined intervention with fibrinogen (4mg/mL) and rFVIIa (4μg/ml) completely reversed the coagulopathy (CT=745 sec, MCF= 69mm) induced by GT or abciximab as evaluated by thromboelastometry. Test run following activation with TF and t-PA showed that abciximab significantly reduced the total area under the elasticity curve from 139×103 to 36×103. Furthermore, addition of fibrinogen dose dependently increased clot stability to 153×103. In contrast, rFVIIa only induced minor increases in clot stability to 50×103. Experiments with Impact R showed that GT and addition of abciximab abolished platelet adhesion and aggregation. No changes were observed following addition of rFVIIa, fibrinogen or their combination. Furthermore, no changes were seen following prolonging the standard duration of the measurement from 2 to 8 mins, neither following increasing the shear from 720/sec to 1500/sec. [Display omitted] ConclusionsOur laboratory studies suggest that urgent reversal of Glanzmann's thrombasthenia and gpIIb/IIIa antagonists may be achieved by combined haemostatic intervention of fibrinogen concentrate and rFVIIa. The mechanisms seem to depend on fibrin polymerisation and improved clot stability rather than shear stress induced impacts on platelet adhesion and aggregation. Future clinical studies are needed to verify the findings. DisclosuresOff Label Use: Fibrinogen is not licensed for use in Glanzmann's thrombasthenia.

Investigation of Interaction Between Enolase and Phosphoglycerate Mutase using Molecular Dynamics Simulation

Two glycolytic enzymes, phosphoglycerate mutase (PGM) and enolase from Saccharomyces cerevisiae have been chosen to detect complex formation between active centers (a/c), using molecular dynamics simulation. Enzymes have been separated by 10 Å distance and placed in a water box of size 173 × 173 × 173 Å. Three different orientations where a/c of PGM and enolase were positioned toward each other have been used for investigation. The two initial 3-phosphoglycerate substrates at near active centers of initial structure of PGM have been replaced with final 2-phosphoglycerate products. 150mM of NaCl have been added to the system to observe binding activity in the near physiological conditions. Analysis of interaction energies and conformation changes for 3ns simulation indicates that PGM and enolase do show binding affinity between their near active regions. Moreover the similarity between final conformations of the first two orientations with the initial conformation of the third orientation suggests that complex formation between a/c of enzymes is not confined only by discussed orientations. Clear interaction of enolase with C-terminal tail of PGM has been recorded. These results suggest that substrate direct transfer mechanism may exist between enzymes.

O polissacarídeo do Anacardium occidentale L. na fase inflamatória do processo cicatricial de lesões cutâneas

O efeito do polissacarídeo de Anacardium occidentale L. (POLICAJU) foi avaliado na fase inflamatória do processo cicatricial em camundongos (Mus musculus) Swiss (n=90), organizados de acordo com o tratamento empregado: Grupo I (NaCl 150mM), Grupo II (ácido ascórbico 75mg ml-1) e Grupo III (emulsão contendo POLICAJU 150mg ml-1 preparado em ácido ascórbico 75mg ml-1). As lesões cutâneas foram realizadas assepticamente na região torácica dorsal e cada ferida foi tratada em dose única (200µl) imediatamente após a cirurgia. As feridas foram avaliadas diariamente sob o ponto de vista clínico e histopatológico até o 6° dia de pós-operatório (PO). No 5° dia PO, observou-se um menor percentual de edema e hiperemia no Grupo III em relação aos grupos controle, ao passo que os valores de área da ferida e do percentual de contração não foram estatisticamente significativos. A avaliação histopatológica do grupo tratado com POLICAJU demonstrou a presença de tecido de granulação fibrovascular no 6° dia PO, enquanto os grupos controle apresentavam tecido de granulação com padrão vascular. O tratamento proposto propiciou sinais flogísticos menos acentuados (edema e hiperemia) durante o período inflamatório, compatível com o processo de reparação mais avançado do ponto de vista histopatológico, sugerindo a possível utilização clínica da emulsão contendo POLICAJU.

Evaluation of the Profile of Thrombin Generation during the Process of Whole Blood Clotting as Assessed by Thromboelastography.

The evaluation of thrombin generation during blood coagulation has become the preferred clinically relevant approach to assess the global integrity of this complex process. In order to give the full picture of the physiology of this multifactorial reaction, all cellular and soluble plasma participants to this reaction have to be present in their natural whole blood environment. Whole blood thrombin generation tests are best suited for this purpose but are somewhat cumbersome for use in real time clinical practice. Thrombelastography has long been recognized as a clinically useful technology for assessment of whole blood coagulation. The purpose of this study was to evaluate the possibility of linking the tracing generated by clotting of whole blood in a Thrombelastograph® (TEG®) Hemostasis System with the generation of thrombin as assessed by thrombin/antithrombin complex (TAT). 320μL of citrated whole blood (3.2% buffered citrate and 1000μg/ml of corn trypsin inhibitor; 1 part to 9 parts of blood) from volunteers was clotted in the presence of 20μL CaCl2 (200mM) and 20μL of Innovin (1/100 diluted in PBS/albumin 4%, pH 7.4). Clotting was monitored with 8 channels of the TEG® system. At different time points (4 to 40 min) the whole blood TEG® reaction cups were put in 2640μL of cold quenching solution (EDTA 50mM, L-benzamidine 10mM, NaCl 150mM, HEPES 2mM, pH 7.4, and 10μL of 10mM D-Phe-Pro-Arg CH2Cl chloromethylketone diluted in 0.01M HCl), vortexed, and centrifuged, and the supernatants were kept at −80°C until assayed for TAT by ELISA. Total Thrombus Generation (TTG) was calculated from the first derivative of the TEG® waveform and was compared to the thrombin generation as measured by TAT as described above. The two vectors of values - the TAT thrombin generation data and the corresponding TEG® TTG - were analyzed using Pearson correlation coefficients and linear, non-linear, and natural log (Ln) transformation of TAT values for least-squares goodness-of-fit curves. The best least-squares fit is the exponential curve (Fig. 1) and its corresponding natural log transformation of TAT value as shown by Fig. 2, with R2 = 0.8787 (P < 0.0001). The prediction equation for TAT thrombin generation is y = 3122.7e0.0005x, where x is the TEG® TTG variable (P < 0.0001). Linearizing using the natural log of the TAT thrombin generation variable (Fig. 2), produces the same R2 as the exponential curve. The prediction equation is y = 8.0465 + 0.0005x, significant at P < 0.0001, where y is the TAT thrombin generation in the Ln transformation variable and x is the TEG® TTG variable. The high magnitude of R2 and the high significance of the prediction equation demonstrate the high efficacy of the prediction of TAT thrombin generation using the TEG® TTG. [Display omitted] [Display omitted]

Distribuição da matéria seca e composição química das raízes, caule e folhas de goiabeira submetida a estresse salino

Um experimento em casa de vegetação foi conduzido com o objetivo de estudar a resposta de plantas de goiabeira (Psidium guajavaL.) a diferentes concentrações de NaCl. As plantas foram cultivadas em solução nutritiva, com 0, 25, 50, 75, 100, 125 e 150 mM de NaCl, e colhidas com 30 e 50 dias após estresse, num delineamento inteiramente casualizado. Foram determinadas: matéria seca das raízes e parte aérea; e porcentagens de Na, Cl, K, Ca e Mg nas raízes, caule e folhas. O estresse salino reduziu a matéria seca total, e a relação parte aérea/raiz foi reduzida. O estresse causou um aumento nos níveis dos íons Na+ e Cl-, com maior acúmulo nas folhas, seguido das raízes. Os teores de Ca foram estáveis nas raízes, mas decresceram no caule e folhas. Os teores de K foram reduzidos, particularmente nas folhas. Ao contrário, os teores de Mg não foram afetados pela salinidade no caule e nas raízes, havendo, entretanto, redução nas folhas. O teor de Na apresentou relação direta com o de Cl e inversa com o de K nas raízes e folhas. O teor de Ca não variou com o de Na nas folhas, mas apresentou uma relação inversa nas raízes. O teor de Mg não variou com o de Na nas raízes e folhas.