Novel Alkaloids from Marine Actinobacteria: Discovery and Characterization

The marine environment is an excellent resource for natural products with therapeutic potential. Its microbial inhabitants, often associated with other marine organisms, are specialized in the synthesis of bioactive secondary metabolites. Similar to their terrestrial counterparts, marine Actinobacteria are a prevalent source of these natural products. Here, we discuss 77 newly discovered alkaloids produced by such marine Actinobacteria between 2017 and mid-2021, as well as the strategies employed in their elucidation. While 12 different classes of alkaloids were unraveled, indoles, diketopiperazines, glutarimides, indolizidines, and pyrroles were most dominant. Discoveries were mainly based on experimental approaches where microbial extracts were analyzed in relation to novel compounds. Although such experimental procedures have proven useful in the past, the methodologies need adaptations to limit the chance of compound rediscovery. On the other hand, genome mining provides a different angle for natural product discovery. While the technology is still relatively young compared to experimental screening, significant improvement has been made in recent years. Together with synthetic biology tools, both genome mining and extract screening provide excellent opportunities for continued drug discovery from marine Actinobacteria.     

The water vapor sorption mechanism and its hysteresis in wood: the water/void mixture postulate

A new method of wood moisture sorption analysis is presented using sorption isotherms of a series of mildly heat-treated specimens with varied and known elemental composition. This method allows the determination of the occupancy of accessible sorption sites in wood as a function of relative humidity h, θ(h) ≈ h, found in agreement with the literature data on the non-freezing water occupancy of hydroxyl groups for h < 0.9. Complementary sorption isotherm shape analysis identifies an empirical power law occupancy function, θ(h) = h ^α , α ≈ 0.73, which is close to the former two determinations in the same humidity range. The validity of widely accepted surface sorption theories for wood with a strongly bound primary layer and loosely bound secondary layers is disproven. To explain the found occupancy function, θ(h) ≈ h, a near-ideal liquid mixture of moisture and polar dynamic microvoids in the cell wall substance is postulated. The power law occupancy function is used to calculate the humidity-dependent number of sorption sites in adsorption/desorption isotherms to show that (1) the number of sorption sites from the adsorption line monotonically increases with increasing humidity—argued to represent the equilibrium number of sorption sites at each humidity, and (2) the number of sorption sites from the desorption line fails to fully return to that of the (equilibrium) adsorption line. Hysteresis is quantitatively explained as the result of non-equilibrium excess sorption sites being occupied according to the occupancy law. The relaxation of non-equilibrium excess sorption sites is satisfactorily modeled by a first-order rate equation. Applying the analysis to study mild thermal modification of moisture sorption isotherms revealed that (1) moisture contents decrease directly linear to the removed amount of sorption sites at all humidity <0.95, and (2) the absolute hysteresis is nearly unaffected as a result of counter-acting effects of the reduced number of sorption sites and reduced amount of relaxation.     

Vergleichende Untersuchungen der Mineralstoffgehalte geschädigter und nicht geschädigter Rotbuchen (Fagus sylvatica) auf Muschelkalkstandorten im Saarland

Comperative study of the mineral content of damaged and visibly healthy red beeches (Fagus sylvatica) on limestone soils in the Saarland 50-80 years old red beeches from a limestone region in the Saarland, differentiated in apparently healthy and diseased trees, were investigated on their mineral content beginning with the roots over stem up to the leaves. Macro- and microelement contents were different between healthy and diseased trees. Magnesium content in the roots of the latter was markedly reduced. Also zinc and phosphorous were decreased in the damaged trees, while manganese was partially accumulated. On the other side, a high sulphur content was observed in the crown area of both types of trees.     

Energetics of load carrying in Nepalese porters

Nepalese porters routinely carry head-supported loads equal to 100 to 200% of their body weight (Mb) for many days up and down steep mountain footpaths at high altitudes. Previous studies have shown that African women carry head-supported loads of up to 60% of their Mb far more economically than army recruits carrying equivalent loads in backpacks. Here we show that Nepalese porters carry heavier loads even more economically than African women. Female Nepalese porters, for example, carry on average loads that are 10% of their Mb heavier than the maximum loads carried by the African women, yet do so at a 25% smaller metabolic cost.     

Does nectar reward affect rarity and extinction probabilities of orchid species? An assessment using historical records from Belgium and the Netherlands

The long-term persistence of plant populations may partly depend on pollination processes and seed production. The exact role of pollination in determining plant population viability still remains largely unclear. Orchid species have often been shown to be pollinator limited. Especially, non-rewarding species are characterized by infrequent pollinator visits and low seed set. Therefore, it can be hypothesized that, if overall population fitness of orchid species is influenced by seed production, non-rewarding species should be more prone to local extinction than rewarding species. To study the importance of nectar reward on orchid persistence, we collected historical records about the distribution of 32 orchid species in Flanders (Belgium) and 37 orchid species in the Netherlands. For both regions, present distribution patterns were compared with historical data. Between 1930 (Flanders) or 1950 (the Netherlands) and 2000, 26 (81%) species showed decreases in distribution range in Flanders, eight of which had gone extinct, whereas 29 (78%) species showed declines in distribution area in the Netherlands, five of which went extinct. Contrary to previously reported results, orchid distribution patterns were not related to nectar reward. There was also no significant relationship between nectar reward and extinction for both regions. Orchids typically occurring in wet grasslands and heathlands suffered greater losses than orchids typically confined to forest habitats or calcareous grasslands. We conclude that the production of nectar does not represent a safeguard for local extinction of orchid species. Habitat loss and other deterministic threats associated with habitat fragmentation and deterioration are more important determinants of orchid persistence.     

Comparative genetic characterization of Porcine Circovirus type 2 samples from German wild boar populations

PCV-2 is involved in "postweaning multisystemic wasting syndrome" (PMWS), "porcine dermatitis and nephropathy syndrome" (PDNS), respiratory and reproductive disorders, and thereby plays a crucial role in today's swine production world-wide. The virus is apparently ubiquitous in domestic pigs and has also been demonstrated in wild pigs. Up to now, a characterization of PCV-2 samples from wild pigs, which might help to estimate the possible role of wild pigs as sources of domestic pig infection, has not been carried out. Spleen samples from 16 PCV-2-positive wild pigs from hunting grounds of four regions in Germany were used for the analysis of the viral genome. In one sample, the complete sequence of the genome was determined. In the other, a 742 nucleotide fragment from the highly variable capsid sequence of the ORF2 was sequenced. Analysis of the sequences led to the identification of three PCV-2 strains. One strain, representing 14 of the 16 samples, was closely related with Chinese, but not with German strains. The genome of this strain was shortened by one nucleotide by a deletion close to the end of ORF2. The deletion led to a shift of the stop-codon and to the insertion of a further codon. Two further strains differed in up to 4.7% of nucleotides and up to 10.5% of amino acids (aa). These strains were aligned with clusters of PCV-2 samples from mainly French and German domestic origin.     

ATP release guides neutrophil chemotaxis via P2Y2 and A3 receptors

Cells must amplify external signals to orient and migrate in chemotactic gradient fields. We find that human neutrophils release adenosine triphosphate (ATP) from the leading edge of the cell surface to amplify chemotactic signals and direct cell orientation by feedback through P2Y2 nucleotide receptors. Neutrophils rapidly hydrolyze released ATP to adenosine that then acts via A3-type adenosine receptors, which are recruited to the leading edge, to promote cell migration. Thus, ATP release and autocrine feedback through P2Y2 and A3 receptors provide signal amplification, controlling gradient sensing and migration of neutrophils.     

Stable carbon isotope composition of perirenal adipose tissue fatty acids from Engadine sheep grazing either mountain or lowland pasture

To provide further insights into ruminant lipid digestion and metabolism, and into cis-9,trans-11 18:2 synthesis, 12 growing Engadine lambs grazing either mountain pasture (2,250 m above sea level; n = 6) or lowland pasture (400 m above sea level; n = 6) were studied. Both pastures consisted exclusively of C(3) plants. Before the experiment, all animals grazed a common pasture for 6 wk. Grasses and perirenal adipose tissues of the sheep were analyzed for fatty acids by gas chromatography. Stable C-isotope ratios (δ(13)C values in ‰ vs. the Vienna Pee Dee Belemnite standard) were determined in the composite samples by elemental analysis-isotope ratio mass spectrometry. The δ(13)C of the individual fatty acids were measured by gas chromatography-combustion-isotope ratio mass spectrometry. The δ(13)C value of the entire mountain pasture grass was -27.5‰ (SD 0.31), whereas that of the lowland pasture grass was -30.0‰ (SD 0.07). This difference was reflected in the perirenal adipose tissues of the corresponding sheep (P < 0.05), even though the δ(13)C values were less in the animals than in the grass. The δ(13)C values for cis-9 16:1 and cis-9 18:1 in perirenal fat differed between mountain and lowland lambs (P < 0.05). The 16:0 in the adipose tissue was enriched in (13)C by 5‰ compared with the dietary 16:0, likely as a result of partly endogenous synthesis. The δ(13)C values of cis-9,trans-11 18:2 (cis-9,trans-11 CLA) in the adipose tissue were smaller than those of its dietary precursors, cis-9,cis-12 18:2 and cis-9,cis-12,cis-15 18:3; conversely, the δ(13)C values of trans-11 18:1 were not, suggesting that large proportions of perirenal cis-9,trans-11 18:2 were of endogenous origin and discrimination against (13)C occurred during Δ(9)-desaturation. The same discrimination was indicated by the isotopic shift between 16:0 and cis-9 16:1 in the mountain grazing group. Furthermore, the δ(13)C values of cis-9,trans-11 18:2 were smaller relative to the precursor fatty acids in the mountain lambs compared with the lowland group. This result suggests a reduced extent of biohydrogenation in lambs grazing on mountain grass in comparison with those grazing on lowland grass. This was supported by the smaller cis-9,trans-11 18:2 concentrations in total fatty acids found in the adipose tissues of the lowland lambs (P < 0.001). The results of this study demonstrate that natural differences between δ(13)C values of swards from different pastures and the adipose tissue fatty acids could be used as tracers in studies of lipid metabolism in ruminants.