Disuccinimidyl suberate cross-linked hemoglobin as a novel red blood cell substitute

Disuccinimidyl suberate (DSS) intramolecularly cross-linked hemoglobin (Hb) was developed as a novel red blood cell substitute. A multi-angle laser light scattering detector coupled with size exclusion HPLC was applied to determine the molecular weight of the modified Hb. SDS-PAGE was also used as a complement. It was proved that 83.8% of the product was intramolecularly cross-linked Hb with weight-average molecular weights (Mw) of 67.5 kD, 12% was dimeric Hb with Mw of 146.6 kD, and 4.2% was trimeric Hb with Mw of 306.4 kD. The tetramer structure of the cross-linked Hb was stable as shown in size-exclusion chromatography using a mobile phase containing 1 mol/L MgCI2. Analysis by LC-MS demonstrated that the reaction of DSS with Hb mainly took place between the two a subunits within a Hb molecule, resulting in stabilization of the tetramer structure. However, the cross-linking was not site-specific. The P50 of the cross-linked Hb decreased from 21.8 mmHg to 14.3 mmHg, and the Hill coefficient decreased from 2.22 to 1.41. Result of isoelectric focusing showed that the pi of DSS cross-linked Hb was in the range of 4.6-5.2, similar to that of serum albumin. The safety of DSS cross-linked Hb was favored by animal tests on rats and guinea pigs. Exchange transfusion experiment with DSS cross-linked Hb using rats as a model indicated no pressor effect or other significant side effects. The characteristics and properties of DSS cross-linked Hb were also compared with that of diaspirin cross-linked Hb reported in the literature.

Growth phenology adjusts to seasonal changes in water availability in coexisting evergreen and deciduous mediterranean oaks

Different leaf(evergreen vs.deciduous habit)and xylem(diffuse-vs.ring-porous wood)traits represent contrasting strategies to face seasonal changes in water availability and temperature.However,how contrasting leaf and xylem habits of coexisting tree species affect stem wood formation and tree-ring development remains poorly understood.Here,we investigated the spatio-temporal patterns of wood formation in two deciduous oaks(Quercus faginea and Quercus petraea)and two evergreen oaks(Quercus ilex and Quercus suber)coexisting in seasonally dry Mediterranean forests along an aridity gradient in Spain.We hypothesized that growth responses to drought and intra-and inter-annual growth patterns would differ between functional groups.We simulated intra-and interannual growth using a modified version of the Vaganov-Shashkin(VS)process-based,growth model.The VS model simulations were used to estimate growth changes under a high emission scenario(RCP 8.5)for the current distribution of the study oak species and to forecast their future performance under warm(4.8℃)conditions in the Iberian Peninsula.Our simulations indicate that climate warming would induce a shortening of the ringgrowth season and a reduction of radial growth in evergreen and deciduous Mediterranean oaks,particularly in dry sites from southern and eastern Iberia currently occupied by Q.ilex and Q.faginea.Evergreen oaks may better recover after dry periods than deciduous oaks by resuming growth after the summer drought.Low soil water availability in spring would be more detrimental to growth of deciduous oaks.Process-based growth models should be refined and validated to better forecast changes in tree growth as a function of climate.

Development of cork oak (Quercus suber L.) seedlings in response to tree shelters and mulching in northwestern Tunisia

The need for reforestation in cork oak （Quercus suber L.） areas is challenged by difficulties. Principal among these is herbivory of young plants, vegetative competition, and slow growth rates of cork oak seedlings. We evaluated the early development of cork oak seedlings treated using tree shelters and mulching in northwestern Tunisia. We tested three tree shelter treatments （non-vented, vented, and control） to shield seedlings from animal damage and five mulch types to control competing vegetation （Italian Stone Pine, Lentisk, combination of Italian Stone Pine and Lentisk （organic mulches）, gravel （inorganic mulch） and no mulch）. At the end of the two-year experiment, sheltered seedlings were 89-99% taller than unshelteredseedlings and had higher numbers and lengths of shoot growth flushes. In contrast, both stem diameter growth and dry weight biomass （from samples extracted after two years） were significantly reduced inside tree shelters. Root-to-shoot ratio was not significantly different in sheltered vs. unsheltered seedlings, suggesting that tree shelters do not adversely affect this parameter. Mulching alone did not favour growth, but could be beneficial when combined with tree shelters. The combination of vented tree shelters and gravel mulch was the most effective treatment for promoting diameter, height and stem volume growth.

Abscisic acid induces differential expression of genes involved in wound-induced suberization in postharvest tomato fruit

Fruit wounding occurred at harvest and transportation requires rapid suberization as a major part of the healing process to prevent infection and desiccation. The focus of this work was to explore the mediation of abscisic acid(ABA) on woundinduced suberization and to determine expression profiles of specific genes involved in wound-induced suberization in tomato fruit. The measurements of weight loss and fruit firmness suggested wound-induced suberization started at 2 d after wounding. The suberization process with the accumulation of suberin polyphenolics(SPP) and polyaliphatics(SPA) observed through autofluorescence microscopy and Sudan IV staining was accelerated by ABA. Expressions of SlPAL5 and Sl4 CL involved in the synthesis of SPP reached the highest at 4 and 8 d after wounding following ABA application, respectively. Associated with SPA biosynthesis, SlLACS1 and SlLACS2 showed the most abundant transcripts at 8 and 6 d in ABA group, respectively. Transcript levels including SlKCSs, SlCYP86 B1, SlFAR3, and Sl GPATs were up-regulated at 2 d after wounding by ABA. Activities of polyphenol oxidase and lipoxygenase were also enhanced during wound-induced suberization following ABA application. The results in this study proved that ABA accelerated the wound-induced suberization progress by increasing the transcript levels of relevant genes in postharvest tomato fruit.

Climate-driven variations in productivity reveal adaptive strategies in Iberian cork oak agroforestry systems

Background:Cork oak agroforestry systems(AFS)have been managed for centuries by humans to produce cork and other goods and services and have recently been recognised as an important reservoir for biodiversity improvement and conservation.However,despite having recently been included as a natural habitat of community-wide interest within the EU Habitats Directive,these systems are in a critical situation of decline.Among other factors,they are strongly threatened by climate change,the effects of which are also expected to be particularly severe in the Mediterranean region.In this study,we aimed to evaluate the influence of climate variability by examining primary production indicators and also to analyse whether the geographical location may have a role in the incidence of the adverse effects of climate.Methods:Cork oak AFS were identified in the Forest Map of Spain and the Land use map of Portugal and categorized on the basis of canopy cover.Seasonal climate data from 2001 to 2020 were used to model relationships with climate predictors and proximity to the coast.Hotspot analysis was conducted to identify significant spatial clusters of high-and low-efficiency areas.Results:The responses to the influence of climatic conditions differed among the various cork oak AFS categories,particularly in the forest category,which was less dependent on climate variations.Relative humidity and water availability were the main drivers of net primary production(NPP).Carbon use efficiency(CUE)was limited by relative humidity and spring temperature in open ecosystems.Proximity to the coast proved beneficial,especially in years with adverse weather conditions,but was not a limiting factor for survival of the ecosystem.Finally,the results of the hotspot analysis supported the other findings,highlighting high-efficiency areas close to the coast and cold spots grouped in specific areas or dispersed inland.Conclusions:Canopy plays a key role in the influence of climatic conditions,particularly in forest categories in which a high density seems to generate microclimate conditions.Water availability,both via the soil and air moisture,is the main driver of primary production,reflecting different adaptive strategies.The oceanic atmosphere may act as a buffer in years of extreme drought.

Effect of Amino Acids on Secondary Somatic Embryogenesis of Moroccan Cork Oak (<i>Quercus suber</i>L.) Tree

In the present study, we tested the effect of amino acids on secondary somatic embryogenesis of Moroccan cork oak (Quercus suber L.). Secondary mature and immature somatic embryos were obtained from primary somatic embryos cultured in N30K medium supplemented with nineteen amino acids. Stimulation of embryogenesis was dependent on the type and concentration of amino acid in the medium. Thus, L-glutamine and L-asparagine at 3.42 mM have been proved to be the most favorable for the formation of functional somatic embryos and the induction of secondary somatic embryos.

Macronutrients Effect on Secondary Somatic Embryogenesis of Moroccan Cork Oak (<i>Quercus suber</i>L.)

To define the preliminary embryogenesis culture conditions of Moroccan Cork Oak (Quercus suber L.) in secondary propagation systems, secondary embryos formation from primary embryos were analyzed using seven macronutrient medias: (Chalupa) (BTM), Murashige and Skoog (MS), Schenk and Hildebrant (SH), Schenk and Hildebrant with half content macronutrients (SH ?), full Gamborg (G), Margara (N30K) and Woody Plant Media(WPM). Mature primary embryos at cotyledonal stage of 8 - 10 mm, were placed in each culture medium, and supplemented with 30 g/l of glucose and 7 g/l of agar without PGR. The experimental design consisted of a Petri dish containing three embryos explants. Each one of the seven treatments was composed of ten Petri dishes. Mean number of secondary somatic embryos, clusters and new embryogenic formation on clusters were recorded after 8 weeks, and evaluated by statistical analysis. There were no significant differences (p ≤ 0.05) in clusters and new embryos on clusters formation among evaluated media;but mean number of secondary embryos was significantly higher in N30K (4.37 ± 0.48) compared with control media (1.37 ± 0.15). The morphology of secondary embryos grown in the N30K medium exclusively showed the presence of three embryogenic stages: early cotyledonal with translucide aspect, white opaque, or green, and mature embryos. These results indicate that the medium do influence the morphogenic characteristics of produced embryos. Our finding revealed that secondary somatic embryos produced in N30K medium presented better morphogenic potential, with different stages of embryogenic formation.

Evaluation of Phenotypic Diversity by Use of Variable Analysis Multi of Various Populations of Oak Cork (<i>Quercus suber</i>L.) in Tunisia

Populations of oak cork (Quercus suber L.) originated from different areas in Tunisia were analysed based on morphological characters to assess the overall degree of phenotypic variability, to detect similarities between the genotypes and to evaluate significant forest features. One-way ANOVA analysis shows a significant difference between oak cork populations and demonstrates that parameters measured on leaves show an important degree of variability and permit to discriminate oak cork individuals. Vegetative variables are leaf length, leaf width, petioles length, number of leaf veins, petioles width, spines number, leaf area, length nipples, diameter nipples, weight nipples, and height trees. Diameter trees were scored and subjected to multivariate analysis. Results showed a considerable diversity among local germoplasme of oak cork. Principal component analysis (PCA) revealed that the characters related to the length and width of the sheets and the nipples accounted for a large proportion of the observed variability. Cluster analysis showed a typically continuous phenotypic diversity among oak cork accessions, and little asso-ciations between accessions from same geographic origin. Also, some heterogeneity within accessions that received the same denomination was evidenced. The variability observed between different populations is attached to phenotypic characters which depend primarily on two factors, gene and environment.

Osmotic Regulation and Viability during Storage of Potato Microtubers Obtained in Vitro

Dehydration,viability,proline,sucrose and glucose concentrations were evaluated on green and white microtubers during storage at two different temperatures.After 10 months at 4℃,the green and white microtubers showed shrinkage with a dry weight loss of 3.91%and 3.15%.Both,the green and white microtubers at 4℃ presented an enhanced sprouting after storage.At 23℃,the green microtubers lost the lowest quantity in dry weight(0.8%)and white microtubers lost 2.2%.This behavior is possibly related to the increase in the thickness of the peridermis observed in green microtubers or to the osmotic regulation mediated principally by the observed concentrations of proline and glucose but not sucrose.The best storage conditions for potato microtubers obtained in vitro were at 4℃ for green or white microtubers for up to 10 months with little loss of viability.

Callogenesis of Cork Oak(Quercus suber L.)through In Vitro Culture of Nodes and Internodes

The present study is about in vitro culture of cork oak, through callogenesis from nodes and internodes, withdrawn from seedlings of three months. These latter were obtained after acorns germination on peat. Nodes showed a high capacity for callogenesis and the best rate was obtained on a medium containing Woody Plant Medium (WPM) macronutrients, Murashige and Skoog (MS) micronutrients and vitamins, 4.5 μM of 6-Benzylaminopurine (BAP), 7 μM of 2-naphthalineacetic acid (NAA) and 30 g/L of Sucrose. Calluses are transplanted onto the same mineral solution, with or without growth regulators, and in the 3rd transplanting;small white embryos appear on the surface of calluses.

Vegetation Dynamism in the Mixed Conifer/Broadleaves Species Artificial Stands Crossed by Fire： The Arci Mountain Case Study in Sardinia

Immediate cutting and logging of mixed conifer/broadleaves species in a man-made forest crossed by fire in 2009 in the Arci Mountain in Sardinia can not guarantee as the best post-fire regeneration. The study case reports the differences in the density and in the height of seedlings. The release of damaged Pinus halepensis Mill. stems seems to confirm the tree seeder role of this species.

Effect on membrane transport in the erythrocytes by band 3 cross-linking

Band 3 and glucose transport protein (GluTl) are two kinds of important proteins in the human erythro-cyte membranes. Bis(sulfosuccinimidyl)suberate (BS ), an impermeable cross-linker of band 3, inhibited NO2- transport, showing that anion exchange is affected by the association state of band 3 in the intact erythrocyte membranes. At the same time, the rates of glucose transport of both exit and entry declined. The amount of monomers of band 3 was decreased after treatment of the erythrocytes with BS3, but there was no change in GluTl according to the SDS-PAGE patterns. This demonstrates that band 3 and GluTl would be linkaged together in the erythrocyte membranes for the requirement of rapid and cooperative performance of physiological functions of the membrane proteins.

Field trapping of the flathead oak borer Coroebus undatus （Coleoptera： Buprestidae） with different traps and volatile lures

The flathead oak borer Coroebus undatus E （Coleoptera： Buprestidae） is one of the primary pests of cork oak Quercus suber L. in the Mediterranean region causing great economic losses to the cork industry. Very little is known about its biology and behavior and, so far, no control measures have been established. We present the results of a pilot study aimed to develop an efficient trapping method for monitoring this harmful pest. In a 3-year field study, purple-colored prism traps baited with a mixture of green leaf volatiles （GLVs） from the host have been shown the most effective combination to catch C. undatus adults （solely females） compared to other trap and lure types tested. Wavelength and reflectance measurements revealed that purple traps exhibit reflectance peak values similar to those found in the abdominal and elytral cuticle of both sexes, suggesting the involvement of visual cues for mate location in this species. The data presented are the first to demonstrate captures of adults of the genus Coroebus by an attractant-based trapping method.