The Interaction of Coumarin and Temperature in the Germination of Lettuce Seed

Dormancy was induced in lettuce seed by exposing to coumarin during imbibition. The level of dormancy which is imposed is dependent on the temperature during germination, coumarin being ineffective below 20°C. At 25°C there is marked interaction between coumarin and this temperature, and the response of the seed. It was shown by exposing treated seeds to temperature regimes involving periods at 25°C and 10°C. that the action of coumarin can be explained more readily by concluding that the events which are affected are cell elongation and division. There is also evidence that the coumarin is metabolised more rapidly at 25°C, and that the effectiveness of the coumarin in the experiments cited depends on its presence at the time when embryo growth is initiated. The mode of action of coumarin is unknown.     

Cleavage efficient 2A peptides for high level monoclonal antibody expression in CHO cells

Linking the heavy chain (HC) and light chain (LC) genes required for monoclonal antibodies (mAb) production on a single cassette using 2A peptides allows control of LC and HC ratio and reduces non-expressing cells. Four 2A peptides derived from the foot-and-mouth disease virus (F2A), equine rhinitis A virus (E2A), porcine teschovirus-1 (P2A) and Thosea asigna virus (T2A), respectively, were compared for expression of 3 biosimilar IgG1 mAbs in Chinese hamster ovary (CHO) cell lines. HC and LC were linked by different 2A peptides both in the absence and presence of GSG linkers. Insertion of a furin recognition site upstream of 2A allowed removal of 2A residues that would otherwise be attached to the HC. Different 2A peptides exhibited different cleavage efficiencies that correlated to the mAb expression level. The relative cleavage efficiency of each 2A peptide remains similar for expression of different IgG1 mAbs in different CHO cells. While complete cleavage was not observed for any of the 2A peptides, GSG linkers did enhance the cleavage efficiency and thus the mAb expression level. T2A with the GSG linker (GT2A) exhibited the highest cleavage efficiency and mAb expression level. Stably amplified CHO DG44 pools generated using GT2A had titers 357, 416 and 600 mg/L for the 3 mAbs in shake flask batch cultures. Incomplete cleavage likely resulted in incorrectly processed mAb species and aggregates, which were removed with a chromatin-directed clarification method and protein A purification. The vector and methods presented provide an easy process beneficial for both mAb development and manufacturing.     

Influence de l'âge des grand-parents sur l'apparition des mâles chez le Rotifère Notommata copeus Ehr.

When parental females (♀♀P = mothers) of the rotifer Notommata copeus are placed in light conditions inducing the appearance of mictic females in their offspring (F₁), the age of the parents of these parental females significantly affects the ratio of mictic females in the F₁ generation.

The preparental females (♀♀PP = grandmothers), the parental females and the F₁ females are isolated in a medium changed at each generation and, in the second experimental series, changed daily: the preparental age effect implicates the transmission of substances on two generations.

This influence of the preparental age is rhythmic: the ratio of mictic females in F₁ related to this age varies in a sinusoidal manner. This influence is endogenous: it persists, always in a sinusoidal form, when the medium in which each grandmother is placed is changed daily.

Furthermore, the net reproduction ratio, Ro, does not vary significantly with the preparental age during these experiments.     

Lower Temperatures More Effective than Atmosphere Modification in Controlling Botrytis and Nectria Rots in Stored Apples

A series of small‐scale controlled inoculation experiments has been conducted during 2005–2009 to determine whether temperature and controlled atmosphere (CA) storage conditions affect significantly the incidence of Botrytis cinerea and Neonectria galligena rots of apples and to assess whether CA regimes can be ‘fine‐tuned’ to suppress fungal rotting. The incidence of B. cinerea and N. galligena rots on apple was reduced consistently by storage in lower temperatures (1.5–2°C). In no case was the disease incidence significantly higher than that under air storage conditions. However, the effect of CA conditions on rot development varied greatly from year to year so that overall there were no significant effects of CA conditions on the incidence of rot during storage till the following April. Thus, we can neither fine‐tune CA conditions to reduce rot development nor exploit the advantages of lower storage temperatures for Cox and Bramley for this purpose due to inevitable development of low temperature breakdown. Further research is needed to study the effects of ethylene control technologies and modulated storage temperatures on rot development.     

A Phase Ia Study to Assess the Safety and Immunogenicity of New Malaria Vaccine Candidates ChAd63 CS Administered Alone and with MVA CS

Background

Plasmodium falciparum (P. falciparum) malaria remains a significant cause of mortality and morbidity throughout the world. Development of an effective vaccine would be a key intervention to reduce the considerable social and economic impact of malaria.

Methodology

We conducted a Phase Ia, non-randomized, clinical trial in 24 healthy, malaria-naïve adults of the chimpanzee adenovirus 63 (ChAd63) and modified vaccinia virus Ankara (MVA) replication-deficient viral vectored vaccines encoding the circumsporozoite protein (CS) of P. falciparum.

Results

ChAd63-MVA CS administered in a heterologous prime-boost regime was shown to be safe and immunogenic, inducing high-level T cell responses to CS. With a priming ChAd63 CS dose of 5×10⁹ vp responses peaked at a mean of 1947 SFC/million PBMC (median 1524) measured by ELIspot 7 days after the MVA boost and showed a mixed CD4+/CD8+ phenotype. With a higher priming dose of ChAd63 CS dose 5×10¹⁰ vp T cell responses did not increase (mean 1659 SFC/million PBMC, median 1049). Serum IgG responses to CS were modest and peaked at day 14 post ChAd63 CS (median antibody concentration for all groups at day 14 of 1.3 µg/ml (range 0–11.9), but persisted throughout late follow-up (day 140 median antibody concentration groups 1B & 2B 0.9 µg/ml (range 0–4.7).

Conclusions

ChAd63-MVA is a safe and highly immunogenic delivery platform for the CS antigen in humans which warrants efficacy testing.

Trial Registration

ClinicalTrials.gov {"type":"clinical-trial","attrs":{"text":"NCT01450280","term_id":"NCT01450280"}}NCT01450280     

A benefit-finding intervention for family caregivers of persons with Alzheimer disease: study protocol of a randomized controlled trial

Background

Patients with ST-elevation myocardial infarction (STEMI) not treated with primary or rescue percutaneous coronary intervention (PCI) are at risk for recurrent ischemia, especially when viability in the infarct-area is present. Therefore, an invasive strategy with PCI of the infarct-related coronary artery in patients with viability would reduce the occurrence of a composite end point of death, reinfarction, or unstable angina (UA).

Methods

Patients admitted with an (sub)acute myocardial infarction, who were not treated by primary or rescue PCI, and who were stable during the first 48 hours after the acute event, were screened for the study. Eventually, we randomly assigned 216 patients with viability (demonstrated with low-dose dobutamine echocardiography) to an invasive or a conservative strategy. In the invasive strategy stenting of the infarct-related coronary artery was intended with abciximab as adjunct treatment. Seventy-five (75) patients without viability served as registry group. The primary endpoint was the composite of death from any cause, recurrent myocardial infarction (MI) and unstable angina at one year. As secondary endpoint the need for (repeat) revascularization procedures and anginal status were recorded.

Results

The primary combined endpoint of death, recurrent MI and unstable angina was 7.5% (8/106) in the invasive group and 17.3% (19/110) in the conservative group (Hazard ratio 0.42; 95% confidence interval [CI] 0.18-0.96; p = 0.032). During follow up revascularization-procedures were performed in 6.6% (7/106) in the invasive group and 31.8% (35/110) in the conservative group (Hazard ratio 0.18; 95% CI 0.13-0.43; p < 0.0001). A low rate of recurrent ischemia was found in the non-viable group (5.4%) in comparison to the viable-conservative group (14.5%). (Hazard-ratio 0.35; 95% CI 0.17-1.00; p = 0.051).

Conclusion

We demonstrated that after acute MI (treated with thrombolysis or without reperfusion therapy) patients with viability in the infarct-area benefit from a strategy of early in-hospital stenting of the infarct-related coronary artery. This treatment results in a long-term uneventful clinical course. The study confirmed the low risk of recurrent ischemia in patients without viability.

Trial registration

ClinicalTrials.gov: NCT00149591.     

The chalk-stream environment

Chalk is a very pure and porous form of limestone. Water percolates down through the rock to form aquifers which overflow as springs forming chalk streams which receive little surface runoff. Passage through the rock is relatively slow and smooths out irregularities in rainfall and stabilises the water temperature. As a result the streams tend to have a regular annual hydrograph with a relatively small difference between winter and summer flows and with no spate conditions; and the temperature range in the streams remains closer to the annual mean than in rivers receiving more surface runoff. The water is calcareous with a pH of 7.4–8.0 and an ample supply of plant nutrients. The streams support diverse and productive communities of plants and animals and the restricted temperature range is particularly suitable for the growth of brown trout.     

Abscisic acid as an endogenous component in lettuce fruits,Lactuca sativa L. cv. Grand Rapids. Does it control thermodormancy?

Summary Abscisic acid (ABA) has been unequivocally identified as an endogenous component of seeds of Grand Rapids lettuce. Both free and bound ABA levels were followed during imbibition at various temperatures but no clear role for ABA in the imposition of thermodormancy emerged. Furthermore it was apparent that different seed batches showed different ratios of free to bound ABA and patterns of changes. An unsuccessful attempt was made to identify bound ABA, presumed to be the glucosyl ester.Abbreviations ABA abscisic acid - ABA Me ester methyl abscisate - ECD electron capture detector - FID flame ionisation detector - GC-MS combined gas liquid chromatography, mass spectrometry - GLC gas liquid chromatography - PVP polyvinyl-pyrrolidone - TLC thin layer chromatography     

Possible role of volatile Fatty acids and abscisic Acid in the dormancy of oats

Species of Avena differ markedly in their levels of pre- and post-harvest dormancy. These species offer the opportunity of determining if dormancy is related to the endogenous level of growth inhibitor. Germinability in two species of differing levels of dormancy, common oat Avena sativa L., and wild oat Avena fatua L. was assessed as were the contents of abscisic acid and volatile fatty acids of chain length C₆-C₁₀. In A. sativa which did not possess postharvest dormancy there was no correlation between germination and inhibitor levels but in A. fatua the relationship between the content of fatty acid and dormancy was good. The loss of these fatty acids in dry storage by evaporation could explain after ripening.