Refining Genotypes and Phenotypes in KCNA2-Related Neurological Disorders

Pathogenic variants in KCNA2, encoding for the voltage-gated potassium channel K_v1.2, have been identified as the cause for an evolving spectrum of neurological disorders. Affected individuals show early-onset developmental and epileptic encephalopathy, intellectual disability, and movement disorders resulting from cerebellar dysfunction. In addition, individuals with a milder course of epilepsy, complicated hereditary spastic paraplegia, and episodic ataxia have been reported. By analyzing phenotypic, functional, and genetic data from published reports and novel cases, we refine and further delineate phenotypic as well as functional subgroups of KCNA2-associated disorders. Carriers of variants, leading to complex and mixed channel dysfunction that are associated with a gain- and loss-of-potassium conductance, more often show early developmental abnormalities and an earlier onset of epilepsy compared to individuals with variants resulting in loss- or gain-of-function. We describe seven additional individuals harboring three known and the novel KCNA2 variants p.(Pro407Ala) and p.(Tyr417Cys). The location of variants reported here highlights the importance of the proline(405)–valine(406)–proline(407) (PVP) motif in transmembrane domain S6 as a mutational hotspot. A novel case of self-limited infantile seizures suggests a continuous clinical spectrum of KCNA2-related disorders. Our study provides further insights into the clinical spectrum, genotype–phenotype correlation, variability, and predicted functional impact of KCNA2 variants.     

Chemical Synthesis and Enzymatic Testing of CMP‐Sialic Acid Derivatives

The cycloSal approach has been used in the past for the synthesis of a range of phosphorylated bioconjugates. In those reports, cycloSal nucleotides were allowed to react with different phosphate nucleophiles. With glycopyranosyl phosphates as nucleophiles, diphosphate‐linked sugar nucleotides were formed. Here, cycloSal‐nucleotides were used to prepare monophosphate‐linked sugar nucleotides successfully in high anomeric purity and high chemical yield. The method was successfully used for the synthesis of three nucleotide glycopyranoses as model compounds. The method was then applied to the syntheses of CMP‐N‐acetyl‐neuraminic acids (CMP‐Neu5NAc) and of four derivatives with different modifications at their amino functions (N‐propanoyl, N‐butanoyl, N‐pentanoyl and N‐cyclopropylcarbonyl). The compounds were used for initial enzymatic studies with a bacterial polysialyltransferase (polyST). Surprisingly, the enzyme showed marked differences in terms of utilisation of the four derivatives. The N‐propanoyl, N‐butanoyl, and N‐pentanoyl derivatives were efficiently used in a first transfer with a fluorescently labelled trisialo‐acceptor. However, elongation of the resulting tetrasialo‐acceptors worsened progressively with the size of the N‐acyl chain. The N‐pentanoyl derivative allowed a single transfer, leading to a capped tetramer. The N‐cyclopropylcarbonyl derivative was not transferred.     

Behavior-change trials to assess the feasibility of improving complementary feeding practices and micronutrient intake of infants in rural Bangladesh

This study used simple rapid-assessment techniques to test the feasibility of increasing the consumption of complementary foods by infants by asking mothers to increase meal quantity or frequency or by altering the viscosity/energy density of the food. The feasibility of using micronutrient supplements either added directly to food or administered as liquid drops was also examined. The study was conducted in rural Bangladesh and involved four separate short-term behavioral change trials. Depending on the trial, fieldworkers recruited 30 to 45 infants 6 to 12 months of age. Following recommendations to increase the amount of food provided to infants, the mean intakes from single meals increased from 40 +/- 23 g on day 1 to 64 +/- 30 g on day 7 (p < 0.05). In a second trial, the mean meal frequency increased from 2.2 +/- 1.3 on day 1 to 4.1 +/- 1.3 on day 7 (p < 0.05). Provision of high-energy-density diets, prepared by decreasing viscosity with alpha-amylase or by hand-mashing rice and dhal into a paste before feeding, increased single-meal energy consumption from 54 +/- 35 kcal to 79 +/- 52 kcal or 75 +/- 37 kcal (p < 0.05), respectively. Both types of micronutrient supplements were well accepted and used according to recommendations. In conclusion, it was possible to change short-term child-feeding behaviors to promote increased food intake, mealfrequency, energy density, and micronutrient consumption. Because each of these interventions lasted for only about 1 week, however, the long-term sustainability of these changes is not known. Moreover, the effect of increased feeding of complementary foods on intakes of breastmilk and total daily consumption of energy and nutrients requires further study.     

Alanine Scan of the Peptide Antibiotic Feglymycin: Assessment of Amino Acid Side Chains Contributing to Antimicrobial Activity

The antibiotic feglymycin is a linear 13‐mer peptide synthesized by the bacterium Streptomyces sp. DSM 11171. It mainly consists of the nonproteinogenic amino acids 4‐hydroxyphenylglycine and 3,5‐dihydroxyphenylglycine. An alanine scan of feglymycin was performed by solution‐phase peptide synthesis in order to assess the significance of individual amino acid side chains for biological activity. Hence, 13 peptides were synthesized from di‐ and tripeptide building blocks, and subsequently tested for antibacterial activity against Staphylococcus aureus strains. Furthermore we tested the inhibition of peptidoglycan biosynthesis enzymes MurA and MurC, which are inhibited by feglymycin. Whereas the antibacterial activity is significantly based on the three amino acids D ‐Hpg1, L ‐Hpg5, and L ‐Phe12, the inhibitory activity against MurA and MurC depends mainly on L ‐Asp13. The difference in the position dependence for antibacterial activity and enzyme inhibition suggests multiple molecular targets in the modes of action of feglymycin.     

Spatial scale and social impacts of biofuel production

The prospect of biofuels going ‘mainstream’ has drawn more attention to the social impacts of the production and use of transport biofuels. Since 2007, many media stories have appeared about alleged negative impacts of biofuels, notably the price of food going up or land-grab by plantation developers. These stories stand in stark contrast with the rosy picture painted by some academics involved in the technical development of bioethanol or biodiesel. This paper explores the questions when and why negative social impacts are likely to occur and under what circumstances more positive impacts might be expected. These impacts are discussed for three geographically defined biofuel supply chains; north-north, south-north and south-south. These three systems differ in the spatial scale of production and consumption and with that comes a different distribution of environmental, social and economic impacts. In the case of domestic production and consumption in developed countries, the social impacts are relatively minor and can be mitigated by social policies. Large scale, export-oriented production systems in developing countries could theoretically yield positive social impacts, but this would require on the one hand the tailored design of ‘pro-poor’ social innovations and interventions on the ground and on the other hand a certification of the supply chain feeding into consumer demand for ‘ethical’ fuel. The latent existence of this demand might be significant but recent NGO campaigns have severely undermined the ethical credentials of biofuels. It would require a persistent and collaborative effort to restore the brand value of ‘green’ fuel, an effort which will require better legislation and radically improved monitoring and enforcement practices in countries where the very absence of these has led to, and is still causing, the large scale destruction of habitats that are carbon sinks of global importance. The significant levels of government funding for biofuels stand in strong contrast with the problematic environmental and social governance of international biofuels supply chains. Notwithstanding the ‘must tackle climate change’ rhetoric by policy makers and in policy documents, this suggests that biofuels policy may be primarily driven by other concerns, especially regarding energy security. We argue that policies that are designed for a rather narrowly defined purpose of ‘security of supply’, cannot be realistically expected to yield high social or environmental benefits, and certainly not abroad.     

Phytohormone Profiles Induced by Trichoderma Isolates Correspond with Their Biocontrol and Plant Growth-Promoting Activity on Melon Plants

The application of Trichoderma strains with biocontrol and plant growth-promoting capacities to plant substrates can help reduce the input of chemical pesticides and fertilizers in agriculture. Some Trichoderma isolates can directly affect plant pathogens, but they also are known to influence the phytohormonal network of their host plant, thus leading to an improvement of plant growth and stress tolerance. In this study, we tested whether alterations in the phytohormone signature induced by different Trichoderma isolates correspond with their ability for biocontrol and growth promotion. Four Trichoderma isolates were collected from agricultural soils and were identified as the species Trichoderma harzianum (two isolates), Trichoderma ghanense, and Trichoderma hamatum. Their antagonistic activity against the plant pathogen Fusarium oxysporum f. sp. melonis was tested in vitro, and their plant growth-promoting and biocontrol activity against Fusarium wilt on melon plants was examined in vivo, and compared to that of the commercial strain T. harzianum T-22. Several growth- and defense-related phytohormones were analyzed in the shoots of plants that were root-colonized by the different Trichoderma isolates. An increase in auxin and a decrease in cytokinins and abscisic acid content were induced by the isolates that promoted the plant growth. Principal component analysis (PCA) was used to evaluate the relationship between the plant phenotypic and hormonal variables. PCA pointed to a strong association of auxin induction with plant growth stimulation by Trichoderma. Furthermore, the disease-protectant ability of the Trichoderma strains against F. oxysporum infection seems to be more related to their induced alterations in the content of the hormones abscisic acid, ethylene, and the cytokinin trans-zeatin riboside than to the in vitro antagonism activity against F. oxysporum.     

Constructing the Forced Migrant and the Politics of Space and Place‐making

Mobility is one of the defining concepts of globalization processes. For some migrants, however, mobility is restricted by international and national laws as well as sociopolitical discourses, which regulate the migrant body and her ability to create social relations. Based on interviews in asylum seeker accommodations in Germany, this study illustrates how asylum seekers are spatially constructed and arrested through bureaucratic labeling and assignment to heterotopias and as a discursive location of transience and difference. Those processes freeze the forced migrant in place, in social and semiotic spaces, and position it as a politicized discursive location. The positioning is indicative of monitoring the Other as a symbol of threat to the nation in times of risk. Overall, the study illustrates the tensions between transnational mobility and fixity and the intersections between globalization, communication, social, legal, and political practice, and space/place‐making.     

The effect of biphasic waveform tilt in transvenous atrial defibrillation

Atrial defibrillation can be accomplished using low energy shocks and transvenous catheters. The biphasic waveform tilt required to achieve optimal atrial defibrillation thresholds (ADFTs) is, however, not known. The effect of single capacitor biphasic waveform tilt modification on ADFT was assessed in 20 patients. Following AF induction the defibrillation pulses were delivered between the catheters positioned in the coronary sinus and the right atrium. The single capacitor biphasic waveform shocks, delivered over the same pathways, consisted of 65% tilt (65/65 biphasic waveform) to produce an overall tilt of 88%, or 50% tilt (50/50 biphasic waveform) to produce an overall tilt of 75%. Although 65/65 biphasic waveform delivers more energy, the shorter duration 50/50 biphasic waveform reduced stored energy ADFT 21%, from 1.34 +/- 0.82 J with 65/65 biphasic to 1.06 +/- 0.81 J. These differences were not statistically significant. Nine patients had lower ADFT with 50/50 biphasic waveform while five patients had lower ADFT with 65/65 biphasic waveform. Equivalent reduction in ADFT was seen in the remaining six patients. The ADFT was 0.83 +/- 0.65 J when both tilts were considered. In conclusion, biphasic waveform tilt modification may affect the ADFT in an individual patient. The optimal biphasic waveform for ADFT is not known.