Histological characterization of <Emphasis Type="Italic">Passiflora pohlii</Emphasis> Mast. root tips cryopreserved using the V-Cryo-plate technique

Cryopreservation stands out as the main strategy to ensure safe and cost efficient long-term conservation of plant germplasm, especially for biotechnological materials. However, the injuries associated with the procedure may result in structural damage and low recovery rates after cooling. Histological analysis provides useful information on the effects of osmotic dehydration, LN exposure, and recovery conditions on cellular integrity and tissue organization, allowing the determination of the critical steps of the cryopreservation protocol and, thus, the use of optimized treatments. Passiflora pohlii Mast. (Passifloraceae) is a native species from Brazil with potential agronomic interest. Recent studies showed the presence of saponins in its roots, which presented antioxidant activity. The goal of this work was to develop a cryopreservation technique for root tips of in vitro-derived plants of P. pohlii using the V-Cryo-plate technique and to characterize the anatomical alterations that occurred during the successive steps of the protocol. Root tips were excised from in vitro plants and precultured before adhesion to cryo-plates and then treated for different periods with the plant vitrification solutions PVS2 or PVS3. Treatment with PVS2 for 45 min resulted in higher recovery (79%) when compared with PVS3 (43%). The greatest number of adventitious roots per cryopreserved explant was also observed after a 45-min exposure to PVS2. Plasmolysis levels were higher in cortical cells of cryopreserved explants treated with PVS2, while pericycle and central cylinder cells were not damaged after this treatment. Thirty days after rewarming, no plasmolysis could be detected, regardless of the experimental conditions.     

Impact of the erineum strain of <Emphasis Type="Italic">Colomerus vitis</Emphasis> (Acari: Eriophyidae) on the development of plants of grapevine cultivars of Iran

The present experiment was aimed at determining the influence of the grape erineum strain of Colomerus vitis (GEM) (Acari: Eriophyidae) on responses of local grapevine cultivars. GEM was applied at five density levels to each of five cultivars, i.e. Shahani, Sahebi Uroomie, Khalili Bovanat, Rishbaba and Sezdang Ghalat (listed from early to late grape ripening). The experiment was performed in a full factorial design (12 replicates each) and effects of the mite on the relative content of leaf chlorophyll, internode and cane length, leaf area and weight, number and size of the erinea, and percentage of leaves with erinea were investigated. Also mite density on leaves and in buds was assessed. Data were analyzed with a two-way ANOVA followed by Tukey’s test to separate means among treatment levels and cultivars. The relative content of chlorophyll (expressed in Spad units) in infested leaves was reduced along with an increase in mite density and it was shown to be highly significant at the two higher mite density levels for Khalili Bovanat, Rishbaba and Sezdang Ghalat; Shahani and Sahebi Uroomie leaves appeared to be less affected by mite infestation. The highest mite density treatment displayed a strong correlation with weight (positive correlation) and size (negative correlation) of the leaves of four cultivars; leaves of Sahebi Uroomie appeared to be less affected. The reduced internode length was weak in infested plants. Most infested plants produced shorter canes and their lengths appeared to have a strong negative correlation with the highest mite density in four cultivars; canes of Sahebi Uroomie did not appear affected. At the highest mite density, canes of Khalili Bovanat and Sahebi Uroomie displayed the most and the least shortening effects, respectively. The percentage of leaves with erinea, as well as the number of erinea per leaves and the diameter of erinea increased along with the mite population density. The mite densities in buds (April 2014) and on leaves with erinea (in November 2013) were higher at the highest treatment level in the medium-late (Rishbaba) and late ripening (Sezdang Ghalat) cultivars, than in the early and early-medium ripening ones. Almost all data collected in the current experiment allowed the conclusion that Sahebi Uroomie and Shahani were less affected than the other cultivars (Khalili Bovanat, Rishbaba and Sezdang Ghalat).     

Characterization of human placental glycosaminoglycans and regional binding to VAR2CSA in malaria infected erythrocytes

Placental malaria is a serious problem in sub-Saharan Africa. Young women are particular susceptible to contracting this form of malaria during their first or second pregnancy despite previously acquired immunity from past infections. Placental malaria is caused by Plasmodium falciparum parasites expressing VAR2CSA on the erythrocyte surface. This protein adheres to a low-sulfated chondroitin sulfate-A found in placental tissue causing great harm to both mother and developing fetus. In rare cases, the localization of infected erythrocytes to the placenta can even result in the vertical transmission of malaria. In an effort to better understand this infection, chondroitin sulfate was isolated from the cotyledon part of the placenta, which should be accessible for parasite adhesion, as well as two non-accessible parts of the placenta to serve as controls. The placental chondroitin sulfate structures and their VAR2CSA binding were characterized. All portions of human placenta contained sufficient amounts of the appropriate low-sulfated chondroitin sulfate-A to display high-affinity binding to a recombinant truncated VAR2CSA construct, as determined using surface plasmon resonance. The cotyledon is the only placental tissue accessible to parasites in the bloodstream, suggesting it is the primary receptor for parasite infected red blood cells.     

Antiviral activity of <Emphasis Type="Italic">Cleome rosea</Emphasis> extracts from field-grown plants and tissue culture-derived materials against acyclovir-resistant <Emphasis Type="Italic">Herpes simplex</Emphasis> viruses type 1 (ACVr-HSV-1) and type 2 (ACVr-HSV-2)

Extracts from Cleome rosea were investigated for their activity against acyclovir-resistant strains of Herpes simplex type 1 (ACVr-HSV-1) and type 2 (ACVr-HSV-2). Methanolic and acidified (1% (v/v) HCl) methanolic extracts were prepared from field-grown plants and in vitro propagated plants, as well as from calli and cell suspension cultures. The extracts presented low cytotoxicity and caused virus titer reduction above 70%, with different mechanisms of action. Extracts from leaves of field-grown plants inhibited viral infection mainly by affecting the virus particle itself (virucidal effect), while extracts from calli acted mainly on cell receptors. On the other hand, all extracts evaluated affected the virus entry across the cell membrane and the intracellular viral replication at similar percentages, causing reduction on titers in the range of 68–90%. This study validated the potential use of in vitro materials as sources of antiherpetic agents from C. rosea.     

Mycelium of Arbuscular Mycorrhizal fungi (AMF) from different genera: form, function and detection

It is often assumed that all species of arbuscular mycorrhizal fungi (AMF) have the same function because of the ubiquity of the arbuscular mycorrhizal symbiosis and the fact that all AMF occupy the same plant/soil niche. Despite apparent differences in the timing of evolutionary divergence and the morphological characteristics of AMF from the different genera, the majority of studies on these fungi use only species of Glomus. There is increasing evidence, however, that the mechanisms involved in the establishment of a mycorrhiza may differ for species and genera of AMF and influence their subsequent function. The aim of this paper is to highlight the diversity in the form and function of AMF from different genera, knowledge of which is vital in understanding their ecological roles. Potential use of biochemical and molecular approaches to detect AMF in planta and ex planta is also discussed. This revised version was published online in June 2006 with corrections to the Cover Date.     

Radiosensitivity of mammalian cell lines engineered to overexpress cytosolic glutathione peroxidase

Reactive oxygen species are believed to be involved in radiation lethality. Glutathione peroxidase is an intracellular enzyme with antioxidant functions. To determine whether increasing the cellular antioxidant capacity can confer radiation resistance, the effect of overexpression of glutathione peroxidase on radiosensitivity was determined in two different cell types. An expression construct including the bovine cytosolic glutathione peroxidase cDNA was used to overexpress this enzyme in cells of the human lymphoblast cell line Sup-T1 as well as the Chinese hamster ovary cell line AA8. Supplementation of the culture media with 30 nM sodium selenite was included to obtain optimal glutathione peroxidase activity. Northern blot analysis confirmed the presence of the construct mRNA, and a standard coupled spectrophotometric assay demonstrated significantly increased glutathione peroxidase activity in the transfected cell lines. An approximately 8-fold increase was found in the Sup-T1 cells, and an approximately 30-fold increase was obtained in the Chinese hamster ovary AA8 cells. Clonogenic survival was assayed in the overexpressing cells and compared to that in control cells transfected with vector alone. Despite significantly increased glutathione peroxidase activity, no observable radioprotection was conferred in either of the two cell lines studied, indicating that increased glutathione peroxidase activity is insufficient to confer radioresistance in the two cell types examined. These data are discussed in the context of using antioxidants as adjuncts to clinical radiotherapy.     

A critical review of the use of Clara cell secretory protein (CC16) as a biomarker of acute or chronic pulmonary effects

Biomarkers associated with asthma aetiology and exacerbation have been sought to shed light on this multifactorial disease. One candidate is the serum concentration of the Clara cell secretory protein (CC16, sometimes referred to as CC10 or uteroglobin). In this review, we examine serum CC16's relation to asthma aetiology and exacerbation. There is evidence that acute exposures to certain pulmonary irritants can cause a transient increase in serum CC16 levels, and limited evidence also suggests that a transient increase in serum CC16 levels can be caused by a localized pulmonary inflammation. Research also indicates that a transient increase in serum CC16 is not associated with measurable pulmonary damage or impairment of pulmonary function. The biological interpretation of chronic changes in serum CC16 is less clear. Changes in serum CC16 concentrations (either transient or chronic) are not specific to any one agent, disease state, or aetiology. This lack of specificity limits the use of serum CC16 as a biomarker of specific exposures. To date, many of the critical issues that must be understood before serum CC16 levels can have an application as a biomarker of effect or exposure have not been adequately addressed.