Pilot-scale culture ofCoffea arabica in a novel loop fluidised bed reactor

A novel bubble free loop fluidized bed reactor for plant cell cultures was developed and tested usingCoffea arabica as a model cell line. The effects of main operational parameters like morphology and size of inoculum, oxygen supply as well as recirculation of sparingly soluble gases on cell growth and alkaloid production rates in this reactor were studied and the results were compared with standard shake flask experiments. By on-line monitoring of biomass and oxygen uptake rates the main kinetic parameters for cell growth and alkaloid production were evaluated. It was demonstrated that the novel reactor is easy to run and is particularly adequate for measuring kinetic parameters necessary for scale up.     

Photosynthetic responses of Coffea arabica leaves to a short-term high light exposure in relation to N availability

It is known that the coffee (Coffea arabica L.) plant which is originally from shade habitats would have a limited ability to grow under full sun. Previous work has shown that nitrogen fertilisation can reduce the leaf damage when the plants are exposed to high light intensities during several days. In the present work we aimed to study the effects of the high irradiance during the first hours and evaluate the positive contribution of nitrogen fertilisation in the case of short-term exposure to strong light. Young plants (1.5–2 years old) grown in 1.5 kg of a mixed soil were supplemented with a nutrient solution containing 15 mM nitrogen in the form of NH₄NO₃, every 7 days (2N treatment), 15 days (1N treatment) and 45 days (0N treatment). Top mature leaves were exposed to a photosynthetic photon flux density of 1 500 μmol m^?2 s^?1 for a maximal period of 8 h, and changes in photosynthesis and pigment composition were monitored along the period of high light exposure. Photosynthetic capacity, leaf conductance to water vapour, electron transport capacity and maximum carboxylation activity, as well as some leaf fluorescence parameters (minimal fluorescence, photochemical efficiency of PSII and quantum yield of photosynthetic electron transport) were reduced by the stress, with a generally stronger impact observed in the 0N plants. The photochemical quenching was affected only in the 0N plants, while the non-photochemical quenching increased in 2N plants but decreased in the 0N ones. The results showed that 2N plants presented a better initial status of the photosynthetic parameters and of the content of photoprotective pigments. Those plants showed ability to trigger some protective mechanisms, as observed by the tendency to increase the xanthophyll pool content, specially in zeaxanthin and in non-photochemical quenching. Also, protein content presented a tendency to increase after 1.5 h, which was maintained until the end of the high light period. We conclude that nitrogen availability is a key factor in the acclimation process to high light.     

Lowering the alemtuzumab dose in reduced intensity conditioning allogeneic hematopoietic cell transplantation is associated with a favorable early intense natural killer cell recovery

Background aimsThe anti-CD52 monoclonal antibody alemtuzumab is employed in allogeneic hematopoietic cell transplantation (alloHCT) for the prevention of graft-versus-host disease (GVHD). However, its optimal dosing in this setting has not been determined yet. We compared three different alemtuzumab dose levels in reduced intensity conditioning (RIC) alloHCT with respect to lymphocyte recovery and outcome.MethodsIn 127 consecutive patients with predominantly advanced stage hematologic malignancies, a first alloHCT after RIC was performed, applying a fludarabine-based protocol (in 93% FBM: fludarabine, bis-chloroethyl-nitrosourea [BCNU], and melphalan). For GVHD prophylaxis, cyclosporine and alemtuzumab at three different dose levels (40 mg, 20 mg, 10 mg) were administered. Recovery of the peripheral blood (PB) lymphocyte sub-populations and clinical outcome were determined with regard to the alemtuzumab dose.ResultsNatural killer (NK) cell concentrations in PB around day +30 correlated inversely with the alemtuzumab dose, whereas other PB lymphocyte subtypes remained essentially unaffected by dosing of alemtuzumab. Lower alemtuzumab doses were associated with a tendency toward improved overall survival mainly during the early post-transplantation months. With regard to the PB NK cell concentration around day +30, “early intense NK cell reconstituters” tended to show an overall survival benefit.ConclusionsAn alemtuzumab dose reduction to only 10–20 mg provides sufficient GVHD prophylaxis and supports improved NK cell regeneration early after alloHCT in PB (“NK cell saving effect”), which may have a positive effect on overall survival.