High potential, but low actual, glycine uptake of dominant plant species in three Australian land-use types with intermediate N availability

The traditional view of the nitrogen (N) cycle has been challenged since the discovery that plants can compete with microbes for low molecular weight (LMW) organic N. Despite a number of studies that have shown LMW organic N uptake by plants, there remains a debate on the overall ecological relevance of LMW organic N uptake by plants across ecosystems with different N availabilities. We here report patterns of glycine N uptake by plants from three different Australian land-use types with intermediate N availability and low inherent glycine concentrations in the soil. Using ¹⁵N labeled tracers, we tested the potential of these plants to acquire glycine in ex-situ laboratory experiments and attempted to validate these results in the field by determining actual uptake of glycine by plants directly from the soil. We found in the ex-situ experiments that plants from all three land-use types were able to take up significant amounts of glycine. In contrast, glycine uptake directly from the soil was minimal in all three land-use types and ¹⁵N tracers were largely immobilized in the soil organic N pool. Our study confirms that the potential for LMW organic N uptake by plants is a widespread phenomenon. However, our in-situ experiments show that in the three land-use types tested here plants are inferior competitors for LMW organic N and rely on NH ₄ ⁺ as their main N source. In contrast to several previous studies in arctic, alpine and even temperate ecosystems, our study suggests that in ecosystems with intermediate N availability, mineral N is the plants’ main N source, while LMW organic N is of less ecological relevance to plant N nutrition.     

Two technetium-99m-labeled cholecystokinin-8 (CCK8) peptides for scintigraphic imaging of CCK receptors

A broad spectrum of radiolabeled peptides with high affinity for receptors expressed on tumor cells is currently under preclinical and clinical investigation for scintigraphic imaging and radionuclide therapy. The present paper evaluates two (99m)Tc-labeled forms of the C-terminal octapeptide of cholecystokinin (CCK8): sulfated (s)CCK8, with high affinity for CCK1 and CCK2 receptors, and nonsulfated (ns)CCK8, with high affinity for CCK2 receptors but low affinity for CCK1 receptors. Peptides were conjugated with the bifunctional chelator N-hydroxysuccinimidyl hydrazino niconitate (s-HYNIC). (99m)Tc-labeling, performed in the presence of nicotinic acid and tricine, was highly efficient (approximately 95%) and yielded products with a high specific activity (approximately 700 Ci/mmol) and good stability (approximately 5% release of radiolabel during 16 h incubation in phosphate buffered saline at 37 degrees C). Chinese hamster ovary cells stably expressing the CCK1 receptor (CHO-CCK1 cells) internalized approximately 3% of added (99m)Tc-sCCK8 per confluent well during 2 h at 37 degrees C. Internalization was effectively blocked by excess unlabeled sCCK8. CHO-CCK1 cells did not internalize (99m)Tc-nsCCK8. Displacement of (99m)Tc-sCCK8 and -nsCCK8 by unlabeled CCK-8 (performed at 0 degrees C to prevent internalization) revealed 50% inhibitory concentrations (IC(50)) of 8 nM and >1 microM, respectively. CHO-CCK2 cells internalized approximately 25% and approximately 5% of added (99m)Tc-sCCK8 and -nsCCK8, respectively. In both cases internalization was blocked by excess unlabeled peptide. IC(50) values for the displacement of (99m)Tc-sCCK8 and -nsCCK8 were 3 nM and 10 nM, respectively. CHO-CCK1 cell-derived tumors present in one flank of athymic mice accumulated 2.0% of injected (99m)Tc-sCCK8 per gram tissue at 1 h postinjection. This value decreased to 0.6% following coinjection with excess unlabeled peptide. Uptake of (99m)Tc-nsCCK8 was low (0.2%) and not did change by excess unlabeled peptide (0.3%). Accumulation of (99m)Tc-sCCK8 and -nsCCK8 by CHO-CCK2 cell-derived tumors (present in the other flank) amounted to 4.2% and 0.6%, respectively. In both cases uptake was significantly reduced by excess unlabeled peptide to 1.0% and 0.4% for sCCK8 and nsCCK8, respectively. Accumulation of (99m)Tc-sCCK8 was also high in pancreas (11.7%), stomach (2.0%), and kidney (2.1%), whereas uptake of (99m)Tc-nsCCK8 was high in stomach (0.7%) and kidney (1.4%). Both radiolabeled peptides showed a rapid blood clearance. In conclusion, these data show that CCK8 analogues can be efficiently labeled with (99m)Tc using s-HYNIC as chelator and nicotinic acid/tricine as coligand system without compromising receptor binding. Furthermore, the present study demonstrates that CCK1 tumors hardly accumulate (99m)Tc-nsCCK8, CCK2 tumors accumulate 2 times more (99m)Tc-sCCK8 than CCK1 tumors, and CCK2 tumors accumulate 15 times more (99m)Tc-sCCK8 than (99m)Tc-nsCCK8. Although accumulation in some nontarget organs was also higher with (99m)Tc-sCCK8, this may not reflect the human situation due to a different receptor expression pattern in humans as compared to mice. Therefore, further studies are warranted to investigate the possible use of (99m)Tc-sCCK8 for scintigraphic imaging of CCK receptor-positive tumors in humans.     

Binding of Two Intrinsically Disordered Peptides to a Multi-Specific Protein: A Combined Monte Carlo and Molecular Dynamics Study

The unique ability of intrinsically disordered proteins (IDPs) to fold upon binding to partner molecules makes them functionally well-suited for cellular communication networks. For example, the folding-binding of different IDP sequences onto the same surface of an ordered protein provides a mechanism for signaling in a many-to-one manner. Here, we study the molecular details of this signaling mechanism by applying both Molecular Dynamics and Monte Carlo methods to S100B, a calcium-modulated homodimeric protein, and two of its IDP targets, p53 and TRTK-12. Despite adopting somewhat different conformations in complex with S100B and showing no apparent sequence similarity, the two IDP targets associate in virtually the same manner. As free chains, both target sequences remain flexible and sample their respective bound, natively -helical states to a small extent. Association occurs through an intermediate state in the periphery of the S100B binding pocket, stabilized by nonnative interactions which are either hydrophobic or electrostatic in nature. Our results highlight the importance of overall physical properties of IDP segments, such as net charge or presence of strongly hydrophobic amino acids, for molecular recognition via coupled folding-binding.     

Oil palm and rubber expansion facilitates earthworm invasion in Indonesia

Deforestation, plantation expansion and other human activities in tropical ecosystems are often associated with biological invasions. These processes have been studied for above-ground organisms, but associated changes below the ground have received little attention. We surveyed rainforest and plantation systems in Jambi province, Sumatra, Indonesia, to investigate effects of land-use change on the diversity and abundance of earthworms—a major group of soil-ecosystem engineers that often is associated with human activities. Density and biomass of earthworms increased 4—30-fold in oil palm and rubber monoculture plantations compared to rainforest. Despite much higher abundance, earthworm communities in plantations were less diverse and dominated by the peregrine morphospecies Pontoscolex corethrurus, often recorded as invasive. Considering the high deforestation rate in Indonesia, invasive earthworms are expected to dominate soil communities across the region in the near future, in lieu of native soil biodiversity. Ecologically-friendly management approaches, increasing structural habitat complexity and plant diversity, may foster beneficial effects of invasive earthworms on plant growth while mitigating negative effects on below-ground biodiversity and the functioning of the native soil animal community.

     

Effect of knee joint cooling on the electromyographic activity of lower extremity muscles during a plyometric exercise

During sporting events, injured athletes often return to competition after icing because of the reduction in pain. Although some controversy exists, several studies suggest that cryotherapy causes a decrease in muscle activity, which may lead to a higher risk of injury upon return to play. The purpose of this study was to investigate the effect of a 20-min knee joint cryotherapy application on the electromyographic activity of leg muscles during a single-leg drop jump in twenty healthy subjects, randomly assigned to an experimental and a control group. After the pre-tests, a crushed-ice bag was applied to the knee joint of the experimental group subjects for 20 min, while the control group subjects rested for 20 min. All subjects were retested immediately after this period and retested again after another 20 min of rest. Average electromyographic activity and ground contact time were calculated for the pre- and post-test sessions. Decreases in electromyographic activity of the lower extremity musculature were found in pre-activation, eccentric (braking), and concentric (push-off) phases immediately after the icing, and after 20 min of rest. The results lend support to the suggestion that cryotherapy during sporting events may place the individuals in a vulnerable position.     

Spatio-temporal variation in post-recovery dynamics in a large Peregrine Falcon (Falco peregrinus) population in the Jura mountains 2000–2020†

After spectacular population crashes in the 1960 and 1970s, Peregrine Falcon Falco peregrinus populations recovered worldwide, and in the 2000s many study populations were back to normal. However, post-recovery trends have been documented less well. We combined three long-term population studies covering the entire Jura mountain range (16 304 km²) to examine spatio-temporal variation in population dynamics during 2000–2020 in 420 known nesting sites in five regions: Ain, Jura and Doubs in France, and Jura Sud and Jura Nord in Switzerland. Every year about 60% of all sites were surveyed at least once, and this proportion increased over time. We used Bayesian occupancy modelling to investigate site persistence and colonization probabilities and correct status and trend estimates for bias due to non-random site coverage. Based on the average of two models correcting for preferential sampling, we estimated that population size peaked in 2002 (Jura), 2005 (Doubs), 2007 (Ain, Jura Nord) and in 2007 for the Jura as a whole. Thereafter, the population declined significantly in four regions and in the Jura as a whole, with the strongest decline found in the Jura Nord ( –38%, 95% credible interval –34%/–41%) during 2007–2020. Thus, we found considerable spatial heterogeneity in post-recovery trends and widespread post-recovery declines. Reasons for the latter may include Eagle Owl Bubo bubo predation, illegal persecution and human disturbance. Our results illustrate how quickly positive population trends may be reversed, such that continuous long-term monitoring even for species seemingly ‘out of danger’ remains invaluable. Our study also emphasizes the importance of correcting for both coverage bias in general and preferential sampling in particular when assessing population trends in studies where not every territory can be surveyed in all years.     

A challenging insight on the structural unit 1 of molluscan Rapana venosa hemocyanin

Hemocyanins of mollusks are high molecular mass glycoproteins with a complex quaternary structure which still remains to be defined in detail for most of its species as far as number, spatial distribution and interactions of their structural units is concerned. In the present study, we isolated the functional units of the structural subunit RvH1 of Rapana venosa hemocyanin, combining enzymatic and non-enzymatic methods. Our results suggest that Hc's carbohydrate moieties play a basic role in the organization of the structural units, resulting from post-translational polymerization of the 50 kDa functional units and involving sugar moieties that link between them.