Amelioration of human osteoarthritis symptoms with topical ‘biotherapeutics’: a phase I human trial

Osteoarthritis (OA) treatments presently rely on analgesics, which manage pain but fail to restore imbalances between catabolic and anabolic processes that underlie OA pathogenesis. Recently, biologic (biotherapeutic) drugs, which alter the activity of catabolic agents such as nitric oxide and inflammatory cytokines in ways, allowing tissue regeneration, were evaluated for efficacy in OA treatment. These studies failed to demonstrate dramatic abatement of OA symptoms by these drugs, but suggested strategies by which biologic agents might be used to treat OA. The present review summarizes current understanding of OA pathogenesis and evolving treatments. Preliminary evaluations of a novel biotherapeutic strategy are presented here. Twenty OA patients receiving sour topical cherry seed extract (SCE), an inducer of heme oxygenase-1 (HO-1), a major physiological protectant against oxidative stress exhibited significantly decreased joint pain and activation of CD4+ T cells expressing inflammatory cytokines (p < 0.05), significantly decreased peripheral blood C-reactive protein (CRP), and increased leukocyte HO-1 (p < 0.05) in comparison with ten placebo-treated patients. SCE inhibits joint-damaging inflammatory mediator production. This agent therefore meets the main criterion for classification as a “biotherapeutic,” or “biologic” agent. The negligible toxicity and low cost of such materials make them promising contributors to OA treatment, sustainable within resource limitations of a wide range of patients.     

Increased growth in elevated [CO2]: an early,short-term response?

Saplings of four clones of Sitka spruce and cherry were grown for three and two growing seasons, respectively, in open top chambers at two CO₂ concentrations (≈ 350 and ≈ 700 μmol mol^–1) to determine whether the increase in total biomass brought about by enhanced [CO₂] is a result of a transient or persistent effect in nonlimiting conditions. Classical growth analysis was applied to both species and mean current relative growth rate of total dry mass (R_T) and leaf dry mass (R_L), and period relative growth rate of total dry mass ( ) and leaf dry mass ( ) were calculated. Sitka spruce saplings and cherry seedlings showed a positive growth response to elevated [CO₂], and at the end of the experiments both species were ≈ 40% larger in elevated [CO₂] than in ambient [CO₂]. As a result, the period mean and were significantly higher in elevated [CO₂]. The differences in plant dry mass at the end of the experiments were a consequence of the more rapid growth in the early phase of exposure to elevated [CO₂]. After this initial phase mean R_T and R_L were similar or even lower in elevated [CO₂] than in ambient [CO₂]. NAR of both species was much higher in elevated [CO₂], whereas both LAR, SLA, and LMR showed the opposite trend. The higher LAR and SLA of plants in ambient [CO₂] contributed to a compensation by which they maintained R_T similar to that of elevated [CO₂] saplings despite lower NAR and photosynthetic rate. However, when the same size the trees were similar amongst the [CO₂] treatments, indicating that one of the main effect of elevated [CO₂] on tree growth is to speed-up early development in all aspects.     

Differential response of four tree species to ozone-induced acceleration of foliar senescence

Ozone (O₃)-induced accelerated senescence of leaves was measured in four tree species: black cherry ( Prunus serotina ), hybrid poplar ( Populus maximowizii x trichocarpa , clone 245), northern red oak ( Quercus rubra ) and sugar maple ( Acer saccharum ). Seedlings or ramets of the four species were subjected to chronic O₃ exposures and designated leaves harvested periodically from emergence to senescence. Gas exchange was analysed, and concentrations of total soluble protein and ribulose-1,5-bisphosphate carboxylase/oxygenase were measured as indicators of leaf senescence. Total antioxidant potential and ascorbate peroxidase and glutathione reductase activities also were determined. Black cherry and hybrid poplar exhibited O₃-induced accelerated leaf senescence, whereas sugar maple and northern red oak did not. When the O₃ effects were related to cumulative uptake of the gas, black cherry was the most sensitive of the four species. Although hybrid poplar exhibited similar symptoms of O₃-induced accelerated senescence after the same exposure period as did black cherry, this species took up much greater quantities of O₃ to achieve the same response. The O₃-induced increase in glutathione reductase activity in hybrid poplar was consistent with the capacity of this species to take up high concentrations of the gas. Relative tolerance of northern red oak and sugar maple could be explained only in part by lower cumulative O₃ uptake and lower rate of uptake. Sugar maple had the highest antioxidant potential of all four species, which may have contributed to O₃ tolerance of this species. Ascorbate peroxidase activity, when expressed on a fresh weight basis, could not account for differential sensitivity among the four species.     

Acclimatizing micropropagated black cherry by comparison with half-sib seedlings

Black cherry ( Prunus serotina Ehrh.) plantlets were cultured in vitro, transferred to potting mixture and evaluated for 8 weeks for acclimatization to a varying, but controlled environment. Whole plantlet growth and water relations were monitored and compared to seedlings of comparable size of the same maternal genotype grown under different conditions, but given the same pretest environment. At one week ex vitro. gravimetrically determined leaf conductance of plantlets was high, but became closer to that of seedlings in both magnitude and diurnal pattern as acclimatization progressed. By 8 weeks, leaf conductance of plantlets was nearly identical to that of seedlings, but the xylem water potential of the plantlets was significantly less. Logarithmic regressions of shoot vs root dry weight indicated that seedlings were allocating twice as much dry matter to shoot than to root growth compared to 4- and 8-week plantlets. Over the same period, stomatal densities of both seedlings and plantlets decreased and stomatal pore lengths increased. Multiple adventitious roots of plantlets emerged from a single site just below the root collar whereas secondary or lateral roots of seedlings originated acropetally along the central root axis. Leaf conductance of plantlets at 8 weeks was similar to that of seedlings indicating satisfactory acclimatization. The larger relative root growth rate of plantlets compared to the shoot, however, was associated with lower stem xylem water potential. The anomaly may be a consequence of the method of in vitro root formation. Plantlet growth rates were lower than growth rates of seedlings and their leaf area was correspondingly less.     

Identification and Genomic Variability of Prune dwarf virus Variants Infecting Stone Fruit Trees in Turkey

Prune dwarf virus (PDV), a pathogen of stone fruit trees worldwide, shows a great variability in its biological, serological and molecular properties. The coat protein sequences of ten PDV variants from different stone fruit tree species in Turkey were determined. The sequence for each variant was 657 nucleotides in length. The phylogenetic analyses of the sequences of the Turkish variants and of additional sequences of other PDV variants from international databases indicate the existence of four groups of PDV variants: one contains cherry variants that differs in group specific conserved amino acids; a second contains one apricot and cherry variants, most members of the group being Turkish variants; a third contains mixed variants from cherry, peach, plum trees and an almond tree; and a fourth contains only variants from almond trees. Based on their hosts, we propose the nomenclature of these groups as cherry I, cherry II, mixed and almond groups, respectively.     

Studies on water transport through the sweet cherry fruit surface: characterizing conductance of the cuticular membrane using pericarp segments

Water conductance of the cuticular membrane (CM) of mature sweet cherry fruit (Prunus avium L. cv. Sam) was investigated by monitoring water loss from segments of the outer pericarp excised from the cheek of the fruit. Segments consisted of epidermis, hypodermis and several cell layers of the mesocarp. Segments were mounted in stainless-steel diffusion cells with the mesocarp surface in contact with water, while the outer cuticular surface was exposed to dry silica (22 ± 1 °C). Conductance was calculated by dividing the amount of water transpired per unit area and time by the difference in water vapour concentration across the segment. Conductance values had a log normal distribution with a median of 1.15 × 10⁻⁴ m s⁻¹ (n=357). Transpiration increased linearly with time. Conductance remained constant and was not affected by metabolic inhibitors (1 mM NaN₃ or 0.1 mM carbonylcyanide m-chlorophenylhydrazone) or thickness of segments (range 0.8–2.8 mm). Storing fruit (up to 42 d, 1 °C) used as a source of segments had no consistent effect on conductance. Conductance of the CM increased from cheek (1.16 ± 0.10 × 10⁻⁴ m s⁻¹) to ventral suture (1.32 ± 0.07 × 10⁻⁴ m s⁻¹) and to stylar end (2.53 ± 0.17 × 10⁻⁴ m s⁻¹). There was a positive relationship (r²=0.066**; n=108) between conductance and stomatal density. From this relationship the cuticular conductance of a hypothetical astomatous CM was estimated to be 0.97 ± 0.09 × 10⁻⁴ m s⁻¹. Removal of epicuticular wax by stripping with cellulose acetate or extracting epicuticular plus cuticular wax by dipping in CHCl₃/methanol increased conductance 3.6- and 48.6-fold, respectively. Water fluxes increased with increasing temperature (range 10–39 °C) and energies of activation, calculated for the temperature range from 10 to 30 °C, were 64.8 ± 5.8 and 22.2 ± 5.0 kJ mol⁻¹ for flux and vapour-concentration-based conductance, respectively. Received: 23 March 2000 / Accepted: 28 July 2000     

Prunus serotina in Italy: a challenging candidate for the national list of priority invasive alien species

Abstract

In accordance with the European regulation on Invasive Alien Species (IAS), the black cherry tree (Prunus serotina Ehrh.) has recently been indicated as one of the 96 species proposed for the development of the national list of priority invasive alien species in Italy. The species, native to North America, is recognised as one of the most harmful IAS in Europe, given its high spread potential and the associated ecological and economic impacts reported in its alien range. Although P. serotina is recognised as a pest within the EU, plants are still available on the market, confirming intentional introduction as a current potential pathway for this species arrival into new areas. Since a comprehensive overview of the main features characterizing the invasive potential of this species is still lacking, we aim to underpin the high priority status of P. serotina as a IAS of main concern in Italy by outlining the main biological features, pathways and impacts of the black cherry in its secondary range. Management measures to be potentially included in specific action plans are also summarised.