Accelerated Nitrate Uptake in Wheat Seedlings: Effects of Ammonium and Nitrite Pretreatments and of 6-Methylpurine and Puromycin

The experiments reported herein had two objectives. One was to determine if the slow rate of nitrate uptake which occurs upon initial exposure of nitrogen-depleted wheat (Triticum vulgare cv. Knox) plants to nitrate was the result of insufficient reduced nitrogen. The second was to determine the impact of restrictions in ribonucleic acid or protein synthesis on both nitrate uptake and nitrate reduction. Pretreatments of 14-day-old seedlings for a few hours in ammonium or nitrite did not result in an enhancement of the initial slow rate of nitrate uptake. Growth for two additional weeks in ammonium also failed to eliminate the induction period. The evidence indicates that the presence of nitrate, rather than a product of its reduction, was required to initiate development of the accelerated rate of nitrate uptake. Puromycin (400 μg ml^?1) and 6-methylpurine (0.5 mM) prevented development of the accelerated phase of nitrate uptake. With both compounds, the relative restriction of nitrate uptake was greater than that of nitrate reduction as revealed by incorporation of ¹⁵N from labeled nitrate into reduced forms. The proportion of reduction which occurred in the root system under the imposed treatments could not be delineated precisely, preventing an unequivocal determination of the extent to which the two processes are coupled in the root system. The data nevertheless indicate nitrate reduction was closely associated with nitrate uptake. Accumulation of nitrate in the shoots was markedly restricted in presence of 6 methylpurine. This effect most likely was a result of a severe restriction in the translocation of nitrate into the xylem, rather than an increase in the reduction rate in the shoots.     

Trans‐equatorial migration,staging sites and wintering area of Sabine’s Gulls Larus sabini in the Atlantic Ocean

The migrations and winter distributions of most seabirds, particularly small pelagic species, remain poorly understood despite their potential as indicators of marine ecosystem health. Here we report the use of miniature archival light loggers (geolocators) to track the annual migration of Sabine’s Gull Larus sabini, a small (c. 200 g) Arctic‐breeding larid. We describe their migratory routes and identify previously unknown staging sites in the Atlantic Ocean, as well as their main Atlantic wintering area in the southern hemisphere. Sabine’s Gulls breeding in northeast Greenland displayed an average annual migration of almost 32 000 km (n = 6), with the longest return journey spanning close to 39 000 km (not including local movements at staging sites or within the wintering area). On their southern migration, they spent an average of 45 days in the Bay of Biscay and Iberian Sea, off the coasts of France, Spain and Portugal. They all wintered in close association with the cold waters of the Benguela Upwelling, spending an average of 152 days in that area. On their return north, Sabine’s Gulls staged off the west African coast (Morocco, Mauritania, Senegal), spending on average 19 days at this site. This leg of migration was particularly rapid, birds travelling an average of 813 km/day, assisted by the prevailing winds. Sabine’s Gulls generally followed a similar path on their outbound and return migrations, and did not exhibit the broad figure‐of‐eight pattern (anti clockwise in the southern hemisphere and clockwise in the northern hemisphere) seen in other trans‐equatorial seabirds in the Atlantic and Pacific oceans.     

The Active and the Inactive Form of the hAT1 Receptor Have an Identical Ligand-Binding Environment: An MPA Study on a Constitutively Active Angiotensin II Receptor Mutant

Several models of activation mechanisms were proposed for G protein-coupled receptors (GPCRs), yet no direct methods exist for their elucidation. The availability of constitutively active mutants has given an opportunity to study active receptor conformations within acceptable limits using models such as the angiotensin II type 1 (AT₁)¹ receptor mutant N111G-hAT₁ which displays an important constitutive activity. Recently, by using methionine proximity assay, we showed for the hAT₁ receptor that TMD III, VI, and VII form the ligand-binding pocket of the C-terminal amino acid of an antagonistic AngII analogue. In the present contribution, we investigated whether the same residues would also constitute the ligand-binding contacts in constitutively activated mutant (CAM) receptors. For this purpose, the same Met mutagenesis strategy was carried out on the N111G double mutants. Analysis of 43 receptors mutants in the N111G-hAT₁ series, photolabeled and CNBr digested, showed that there were only subtle structural changes between the wt-receptor and its constitutively active form.     

Mating behavior and genitalic counterparts in tiger beetles (Carabidae: Cicindelinae)

Comparative studies on the structure of genitalia in Pseudoxychila tarsalis Bates and the copulating behavior in 5 species of Cicindela respectively complement similar findings by Freitag [1] on Cicindela spp. and Palmer [4] on P. tarsalis. These strengthen the hypothesis that in tiger beetles the flagellum fits into the spermatheca duct during copulation; that the main function of the flagellum, which is closed at the apex and not connected to the ejaculatory duct, is to open and prepare the lumen of the spermatheca duct for sperm movement from the bursa copulatrix to the spermatheca; and that copulation proceeds in 3 phases: phase 1 in which the lumen of the spermatheca duct is cleared by the flagellum, phase 2 in which the flagellum is withdrawn from the spermatheca duct, and phase 3 in which semen is transferred from the gonopore of the ejaculatory duct to the bursa copulatrix, usually with a spermatophore.     

Prefeeding larval development time is not correlated with egg size in regular echinoids (Strongylocentrotus species)

Summary

We compared prefeeding development times, from fertilized egg to prism larva, for Strongylocentrotus embryos from four clutches of eggs (each from a different species) differing in size. Development times did not vary consistently with egg diameter, and trends among eggs of different sizes varied with stage of development. In some cases, development times for eggs of intermediate diameter (S. franciscanus) were longer than those for larger or smaller eggs. Although mean egg diameters in clutches ranged from 84 μm (S. purpuratus) to 162 μm (S. pallidus), differences in development time to the last embryonic stage (prism) were very small. We conclude that the inverse relationship between parental investment in offspring and premetamorphic development time in echinoids depends only on the functional consequences of reduced size of feeding larval stages: effects of egg size on prefeeding development time are not evident.     

A new diverse shark fauna from the Wordian (Middle Permian) Khuff Formation in the interior Haushi‐Huqf area,Sultanate of Oman

Abstract: Chondrichthyans are newly reported from the autochthonous Wordian Khuff Formation (middle Permian), cropping out in well‐exposed, low‐palaeolatitude sections in the interior Haushi‐Huqf area of Oman. The shark remains comprise isolated teeth, dermal denticles and fin spines and have been recovered by processing limestone in buffered acetic acid from bulk rock samples. The fauna consists of mainly ctenacanthiform and hybodontiform taxa, identified as Glikmanius cf. myachkovensis, Glikmanius culmenis sp. nov., Omanoselache hendersoni gen. et sp. nov., Omanoselache angiolinii gen. et sp. nov., cf. Omanoselache sp., Reesodus underwoodi gen et sp. nov., Teresodus amplexus gen. et sp. nov., Gunnellodus bellistriatus, Khuffia lenis gen. et sp. nov., Khuffia prolixa gen. et sp. nov. and Euselachii sp. indet. Additional specimens include rare teeth of the lonchidiid cf. ‘Palaeozoic Genus 1’ sp., of the neoselachian Cooleyella cf. fordi and a further indeterminate neoselachian, of an indeterminate petalodont and of the holocephalan Deltodus aff. mercurei and Solenodus cf. crenulatus. Fin spines add a further two taxa, Nemacanthus sp. and Amelacanthus cf. sulcatus, which have neoselachian affinities and therefore an unclear relationship to the recovered teeth. The occurrence of Nemacanthus within this Wordian fauna represents the oldest record of this taxon and its only known occurrence in the Palaeozoic. Of the remaining genera, Glikmanius has previously been recorded from the Wordian, whereas for all the others, this study represents their youngest known stratigraphic occurrence and first occurrence in Guadalupian (middle Permian) strata. This adds significantly to our knowledge of the global diversity of chondrichthyans preceding the end‐Guadalupian biotic crisis. Palaeogeographically, for all taxa, this study represents the first record from the western fringe of the marine Neotethyan basin, and only Cooleyella was previously known from the southern (Gondwanan) part of the Pangaean continental margin.     

Identification of senescent cells in multipotent mesenchymal stromal cell cultures: Current methods and future directions

Regardless of their tissue of origin, multipotent mesenchymal stromal cells (MSCs) are commonly expanded in vitro for several population doublings to achieve a sufficient number of cells for therapy. Prolonged MSC expansion has been shown to result in phenotypical, morphological and gene expression changes in MSCs, which ultimately lead to the state of senescence. The presence of senescent cells in therapeutic MSC batches is undesirable because it reduces their viability, differentiation potential and trophic capabilities. Additionally, senescent cells acquire senescence-activated secretory phenotype, which may not only induce apoptosis in the neighboring host cells following MSC transplantation, but also trigger local inflammatory reactions. This review outlines the current and promising new methodologies for the identification of senescent cells in MSC cultures, with a particular emphasis on non-destructive and label-free methodologies. Technologies allowing identification of individual senescent cells, based on new surface markers, offer potential advantage for targeted senescent cell removal using new-generation senolytic agents, and subsequent production of therapeutic MSC batches fully devoid of senescent cells. Methods or a combination of methods that are non-destructive and label-free, for example, involving cell size and spectroscopic measurements, could be the best way forward because they do not modify the cells of interest, thus maximizing the final output of therapeutic-grade MSC cultures. The further incorporation of machine learning methods has also recently shown promise in facilitating, automating and enhancing the analysis of these measured data.     

Eleven polymorphic microsatellite loci in Lindera benzoin,Lauraceae

Microsatellite markers were developed for studies of the genetic diversity and population substructure of Lindera benzoin, Lauraceae (spicebush). Nuclear microsatellite sequences were obtained from DNA libraries that were enriched for (CA), (GA), (AAG) and (ATG) repeat motifs. From 69 microsatellite sequences, 20 primer sets were developed. Of these, 11 primer pairs resulted in amplified polymorphic loci. In 29 samples of eastern Pennsylvania spicebush plants, the number of microsatellite alleles ranged from two to 16 per locus with observed heterozygosity values ranging from 0.10 to 0.82.