Effects of Acibenzolar-S-methyl on the specific activities of peroxidase, chitinase and phenylalanine ammonia-lyase and phenolic content of host leaves in cucumber-powdery mildew interaction

One of the properties of systemic acquired resistance in plants is its concomitance with the biochemical changes including enhancement of activities of defense-related enzymes. In this study, the effects of Acibenzolar-S-methyl (ACI) on the some of defense responses of the cucumber plants (healthy or inoculated with spore suspension of Podosphaerafusca, the causal agent of cucumber powdery mildew) were surveyed via in vivo tests. Changes of defense responses in ACI-treated cucumber plants, inoculated with pathogen or not, were studied and compared with those of non-treated control plants. Results indicated that specific activity of peroxidase increased significantly in treated plants. Increase in enzyme activity was higher in pathogen-inoculated than non-inoculated plants, thus pathogen attack stress to plant plays a role in enhancement of enzyme activity. Specific activity of phenylalanine ammonialyase showed no changes in ACI-treated non-inoculated plants, but in inoculated plants it Increased due to interaction between ACI treatment and pathogen attack stress. Specific activity of chitinase increased in both inoculated and non-inoculated ACI-treated plants at 24 hours after treatment onwards, and pathogen attack stress did not affect it. Phenolic content of ACI-treated plant tissues, despite of small fluctuations, did not show any definite pattern of changes.     

Evaluation of laboratory-based measures of soil mineral nitrogen and potentially mineralizable nitrogen as predictors of field-based indices of soil nitrogen supply in potato production

Accurate estimation of soil nitrogen (N) supply in the field is required to optimize fertilizer N management and to minimize environmental N losses in humid environments. Laboratory-based measures of N availability were evaluated as predictors of field-based indices of soil N supply within potato production systems. Pre-plant soil samples (0–15 cm) were collected from a series of forty treatments in established potato trials located in Atlantic Canada and Maine, USA. Total plant N uptake at topkill with no fertilizer N applied (PNU_0N), PNU_0N plus soil mineral N to 30 cm depth at harvest and relative yield were considered as field-based indices of soil N supply. The potentially mineralizable N (N₀) was determined by aerobic incubation at 25°C and periodic leaching for 24 weeks. A series of laboratory-based measures of soil N supply were measured in soil samples. Pre-plant soil nitrate or total mineral N at 0–30 cm depth was the best single predictor of PNU_0N (r = 0.67 and 0.71, respectively) and relative yield (r = 0.58 and 0.61). The ultraviolet absorbance of 0.01 M NaHCO₃ extract at 205 nm (NaHCO₃-205) was suitable as a predictor of PNU_0N and relative yield in each growing season, however, the relationship between this parameter and PNU_0N or relative yield varied somewhat among years. A combination of pre-plant mineral N plus N mineralized in the first 2 weeks period of incubation after re-wetting is proposed as a more robust measure of N availability compared with use of mineral N alone.     

Antileishmanial and Cytotoxic Effects of Essential Oil and Methanolic Extract of Myrtus communis L.

Plants used for traditional medicine contain a wide range of substances that can be used to treat various diseases such as infectious diseases. The present study was designed to evaluate the antileishmanial effects of the essential oil and methanolic extract of Myrtus communis against Leishmania tropica on an in vitro model. Antileishmanial effects of essential oil and methanolic extract of M. communis on promastigote forms and their cytotoxic activities against J774 cells were evaluated using MTT assay for 72 hr. In addition, their leishmanicidal activity against amastigote forms was determined in a macrophage model, for 72 hr. Findings showed that the main components of essential oil were α-pinene (24.7%), 1,8-cineole (19.6%), and linalool (12.6%). Findings demonstrated that M. communis, particularly its essential oil, significantly (P<0.05) inhibited the growth rate of promastigote and amastigote forms of L. tropica based on a dose-dependent response. The IC₅₀ values for essential oil and methanolic extract was 8.4 and 28.9 μg/ml against promastigotes, respectively. These values were 11.6 and 40.8 μg/ml against amastigote forms, respectively. Glucantime as control drug also revealed IC₅₀ values of 88.3 and 44.6 μg/ml for promastigotes and amastigotes of L. tropica, respectively. The in vitro assay demonstrated no significant cytotoxicity in J774 cells. However, essential oil indicated a more cytotoxic effect as compared with the methanolic extract of M. communis. The findings of the present study demonstrated that M. communis might be a natural source for production of a new leishmanicidal agent.     

Molecular pathogenesis of sialic acid storage diseases: insight gained from four missense mutations and a putative polymorphism of human sialin

Background information. Free sialic acid storage diseases are caused by mutations of a lysosomal sialic acid transporter called sialin. We showed recently that the milder clinical form, Salla disease, and a related non‐Finish case, are characterized by residual transport, whereas sialin mutants found in lethal infantile cases are inactive. In the present study, we have characterized the molecular effects of a putative polymorphism (M316I) and of four pathogenic mutations associated with either infantile (G127E and R57C) or Salla‐like (G409E) phenotypes, or both (G328E). The transport activity of human sialin was analysed using a novel assay that was based on a construct without the functional lysosomal sorting motif, which is expressed at the plasma membrane. Results. The lysosomal localization of human sialin was not (M316I and G328E) or only partially (R57C, G127E and G409E) affected by the missense mutations. In contrast, all pathogenic mutations abolished transport, whereas the putative M316I polymorphism induced an approx. 5‐fold decrease of sialic acid transport. Conclusions. The molecular effects of the R57C and G127E mutations strengthen the conclusion that the infantile phenotype is caused by loss‐of‐function mutations. On the other hand, the milder severity of the heterozygous G409E patient may reflect an incomplete expression of the splicing mutation present on the second allele. In the case of the G328E mutation, found in the homozygous state in a clinically heterogeneous family, the apparent severity of the transport phenotype suggests that the genetic or environmental factors underlying this clinical heterogeneity might be protective.     

Establishment of callus and cell suspension culture of Scrophularia striata Boiss.: an in vitro approach for acteoside production

In the present study, a protocol was optimized for establishment of callus and cell suspension culture of Scrophularia striata Boiss. as a strategy to obtain an in vitro acteoside producing cell line for the first time. The effects of growth regulators were analyzed to optimize the biomass growth and acteoside production. The stem explant of S. striata was optimum for callus induction. Modified Murashige and Skoog medium supplemented with 0.5 mg/l naphthalene acetic acid + 2.0 mg/l benzyl adenine was the most favorable medium for callus formation with the highest induction rate (100 %), the best callus growth and the highest acteoside content (1.6 μg/g fresh weight). Incompact and rapid growing suspension cells were established in the liquid medium supplemented with 0.5 mg/l naphthalene acetic acid + 2.0 mg/l benzyl adenine. The optimum time of subculture was found to 17–20 days. Acteoside content in the cell suspension was high during exponential growth phase and decreased subsequently at the stationary phase. The maximum content of acteoside (about 14.25 μg/g cell fresh weight) was observed on the 17th day of the cultivation cycle. This study provided an efficient way to further regulation of phenylethanoid glycoside biosynthesis and production of valuable acteoside, a phenylethanoid glycoside, on scale-up in S. striata cell suspension culture.     

Identification of FABP7 in fibroblastic reticular cells of mouse lymph nodes

Fatty acids and their metabolites regulate immune cell function. The present study was undertaken to examine the detailed distribution of fatty acid binding proteins (FABPs), the cytosolic chaperones of fatty acids, in mouse peripheral immune organs. Using immunohistochemistry, FABP7 was localized to the alpha-smooth muscle actin (SMA)⁺ fibroblastic reticular cells, which construct the stromal reticula in the T cell areas of the peripheral lymph nodes and spleen. Immunoelectron microscopy showed that FABP7⁺ cells enclosed the collagen fibers, forming a conduit system, which transport lymph and associated low-molecular-mass proteins. In contrast, FABP5⁺ cells were distributed throughout the lymph node and contained well-developed lysosome and phagocytic materials within the cytoplasm. The mesenteric lymph nodes of FABP7 knockout mice showed normal histological features, but the percentage of CD4⁺ cells was significantly increased compared with that in wild-type mice. These data indicate that FABP7 may be involved in T cell homeostasis, possibly by modulating lipid metabolism in fibroblastic reticular cells within the peripheral lymph nodes.     

EFFECT OF MANIPULATION OF WATER AND NITROGEN SUPPLIES ON THE QUANTITATIVE PHENOLOGY OF LARREA TRIDENTATA (CREOSOTE BUSH) IN THE SONORAN DESERT OF CALIFORNIA

Two years of water and nitrogen augmentation experiments on Larrea tridentata (creosote bush) were carried out in a southern Californian warm desert wash plant community. Treatments consisted of control (C), water (W), water and soil nitrogen (W + SN), and soil nitrogen (SN). Quantitative phenological data and microclimatic measurements were collected prior to the onset of and during the growth period and treatments. Predawn and midday water potentials were lower in nonirrigated than irrigated individuals. Leaf conductance was higher in irrigated than in nonirrigated shrubs, with a maximum difference of 1 cm s^-1 observed in July 1984 under relatively low vapor pressure deficit conditions. Leaf production rates were significantly higher in the irrigated (W and W + SN) treatments than in the nonirrigated (C and SN) treatments in 1984. Addition of soil nitrogen caused no increased in vegetative growth rates in 1984. In 1985, a drier year, there was only minimal growth during the spring and summer growth periods in the nonirrigated treatments, while the W and W + SN treatments resulted in significantly higher leafand shoot growth rates. Growth rates in 1985 were significantly higher in the W + SN treatment than in the W treatment. Reproductive growth was higher in the nonirrigated than the irrigated treatments, with the lowest reproductive activity noted in the W treatment.     

Strategies of enzyme immobilization on nanomatrix supports and their intracellular delivery

Abstract

Enzymes are one of the foundations and regulators for all major biological activities in living bodies. Hence, enormous efforts have been made for enhancing the efficiency of enzymes under different conditions. The use of nanomaterials as novel carriers for enzyme delivery and regulating the activities of enzymes has stimulated significant interests in the field of nano-biotechnology for biomedical applications. Since, all types of nanoparticles (NPs) offer large surface to volume ratios, the use of NPs as enzyme carriers affect the structure, performance, loading efficiency, and the reaction kinetics of enzymes. Hence, the immobilization of enzymes on nanomatrices can be used as a useful approach for direct delivery of therapeutic enzymes to the targeted sites. In other words, NPs can be used as advanced enzyme delivery nanocarriers. In this paper, we present an overview of different binding of enzymes to the nanomaterials as well as different types of nanomatrix supports for immobilization of enzymes. Afterwards, the enzyme immobilization on nanomaterials as a potential system for enzyme delivery has been discussed. Finally, the challenges associated with the enzyme delivery using nano matrices and their future perspective have been discussed.

Communicated by Ramasamy H. Sarma