Influence of HIV-1 Genomic RNA on the Formation of Gag Biomolecular Condensates

Biomolecular condensates (BMCs) play an important role in the replication of a growing number of viruses, but many important mechanistic details remain to be elucidated. Previously, we demonstrated that the pan-retroviral nucleocapsid (NC) and HIV-1 pr55^Gag (Gag) proteins phase separate into condensates, and that HIV-1 protease (PR)-mediated maturation of Gag and Gag-Pol precursor proteins yields self-assembling BMCs that have HIV-1 core architecture. Using biochemical and imaging techniques, we aimed to further characterize the phase separation of HIV-1 Gag by determining which of its intrinsically disordered regions (IDRs) influence the formation of BMCs, and how the HIV-1 viral genomic RNA (gRNA) could influence BMC abundance and size. We found that mutations in the Gag matrix (MA) domain or the NC zinc finger motifs altered condensate number and size in a salt-dependent manner. Gag BMCs were also bimodally influenced by the gRNA, with a condensate-promoting regime at lower protein concentrations and a gel dissolution at higher protein concentrations. Interestingly, incubation of Gag with CD4⁺ T cell nuclear lysates led to the formation of larger BMCs compared to much smaller ones observed in the presence of cytoplasmic lysates. These findings suggest that the composition and properties of Gag-containing BMCs may be altered by differential association of host factors in nuclear and cytosolic compartments during virus assembly. This study significantly advances our understanding of HIV-1 Gag BMC formation and provides a foundation for future therapeutic targeting of virion assembly.     

Starch in prasinophycean algae

Reserve products isolated from three species of prasinophycean algae: Pyramimonas parkeae Norris and Pearson, Pyramimonas amylifera Conrad and Platymonas tetrathele West were compared. Three pieces of experimental evidence indicate that the reserve product of each of the species analyzed is a true starch. (I) There is no essential difference between the light absorption curve for the complex formed between corn starch and iodine and the corresponding curves for the algal products. (2) The β-amylase breakdown limit of these compounds is very close to that of corn starch. (3) The X-ray diffraction diagrams produced by the reserve material from Pyramimonas parkeae and Platymonas tetrathele is a so-called A-spectrum characteristic of cereal starches while the diagram produced by grains from Pyramimonas amylifera corresponds to the B-spectrum typical for tuber starches. The dissimilarity between the X-ray diagrams produced by the starches from the Pyramimonas species suggests that a re-examination of the taxonomy of these species is warranted.     

Holocene pollen diagrams from Greece

Pollen diagrams are presented from three sites in Greece. Two cores from the Strymon Delta in Macedonia are thought to span parts of the period from Classical to Turkish times. A dated core from Kleonai in the Peloponnese has good pollen preservation in the early Neolithic, Bronze Age and Roman periods. A peat bog at Asi Gonia in the White Mountains of Crete has a continuous pollen record for the last 1500 years. The palynological evidence for human impact in various historic periods is assessed in the light of archaeological evidence. The problems of interpretation of pollen cores from Greece are discussed and the complex inter‐relationship between human impact and environmental factors is emphasised.     

Antibiotic purification from fermentation broths by counter-current chromatography: analysis of product purity and yield trade-offs

Counter-current chromatography (CCC) is a low pressure, liquid–liquid chromatographic technique which has proven to be a powerful purification tool for the high-resolution fractionation of a variety of active pharmaceutical compounds. The successful integration of CCC into either existing or new manufacturing processes requires the predictable purification of target compounds from crude, fermentation-derived, feed streams. This work examines the feasibility of CCC for the purification of fermentation-derived erythromycin A (EA) from its structurally and chemically similar analogues. At the laboratory scale, the effect of feed pre-treatment using either clarified, forward extracted (butyl acetate) or back extracted broth on EA separation was investigated. This defined the degree of impurity removal required, i.e. back extracted broth, to ensure a reproducible elution profile of EA during CCC. Optimisation and scale-up of the separation studied the effects of mobile phase flow (2–40 mlmin^–1) and solute loading (0.1–10 g) on the attainable EA purity and yield. The results in all cases demonstrated a high attainable EA purity (>97% w/w) with throughputs up to 0.33 kgday^–1. Secondly, a predictive scale-up model was applied demonstrating, that from knowledge of the solute distribution ratio of EA (K_EA) at the laboratory scale, the EA elution time at the pilot scale could be predicted to within 3–10%, depending upon the solute injection volume. In addition, this study has evaluated a fractionation diagram approach to visually determine the effects of key operational variables on separation performance. This resulted in accurate fraction cut-point determination for a required degree of product purity and yield. Overall, the results show CCC to be a predictable and scaleable separation technique capable of handling real feed streams.     

DNA complexes with polycations useful for delivery of DNA into cells

Generation and physicochemical properties of complexes formed by high-molecular thymus DNA and plasmid DNA with synthetic polymers of (dimethyl amino)ethyl methacrylate, (diethyl amino)ethyl methacrylate, and poly(vinyl amine) were studied in solutions of different ionic strength using low-gradient viscometry, electrophoresis, circular dichroism, spectrophotometry, and dynamic light scattering. The complexes were tested for toxicity with T98G cell cultures. Condensation of DNA was shown to occur when the ratio of charged groups in the polycations and DNA exceeded unity. This condensation manifested itself as an increase in the optical density of DNA solutions. Condensation-associated changes in the dimensions of DNA molecules were determined, and phase diagrams of DNA-polycation systems were analyzed in the presence of NaCl. MTT analysis revealed no toxicity of these complexes.     

The Art of Sorting: Using Venn Diagrams to Learn Science Process Skills

In this article, the author describes activities that have been proven to teach young learners to sort and classify objects that contain more than one attribute. The activities require that students employ the use of sorting hoops and attribute blocks to create Venn diagrams, the assembly of which requires practice. The author also describes crosscurricular uses of these learning tools.     

Observations on graphing paleozoological data: Suggestions for better graphs

Paleozoologists have long used graphs of diverse styles to describe, analyze, and summarize their data. Some of these graphs provide excellent visual representations of complex data and are readily deciphered. Other graph styles require close study to be interpreted. Ease of visual decoding of information contained in a graph – graph perception – varies from graph style to graph style. Historical instances of graphing paleozoological data indicate some difficult to decipher graph styles have been used for at least a century. Graphs with three-dimensions, moiré effects, superimposed lines, or segmented bars, or which demand simultaneous decipherment of position and magnitude, are ill-advised. Temporal trends in data are best graphed following the principle of superposition such that data from old material is graphed at the bottom and data from younger material is graphed at the top of the diagram.     

Floral morphology,micromorphology and palinology of selected Sedum s.l. species (Crassulaceae)

Sedum L. (Crassulaceae) is a large and taxonomically difficult genus whose delimitation and classification are under debate. Due to the controversial results of previous cytological, morphological, and molecular studies, further investigations are needed in order to gain a shared taxonomy of the current recognized species clades. In the present paper, morphological and micromorphological features of 23 selected Mediterranean species of Sedum s.l. – collected from exsiccata or fresh specimens throughout Italy – were investigated, in order to provide additional data toward their classification above species level. In particular, the study focused on flower structure and morphology, floral epidermal surfaces and pollen morphology. The distribution pattern of the examined microcharacters across the species revealed a wide range of variation and different combinations of the single characters. NMDS analysis allowed individuating discrete groups that showed a general consistency with the current systematic delimitation of species groups. Our study also evidenced for the first time the wide morphological variability of nectaries and of the glandular indumentum, not previously investigated in detail. In addition, we proposed the combined use of floral diagrams and floral formulae as valuable tools in studying the variability of flower structure at genus level.     

Fundamental physical cellular constraints drive self‐organization of tissues

Morphogenesis is driven by small cell shape changes that modulate tissue organization. Apical surfaces of proliferating epithelial sheets have been particularly well studied. Currently, it is accepted that a stereotyped distribution of cellular polygons is conserved in proliferating tissues among metazoans. In this work, we challenge these previous findings showing that diverse natural packed tissues have very different polygon distributions. We use Voronoi tessellations as a mathematical framework that predicts this diversity. We demonstrate that Voronoi tessellations and the very different tissues analysed share an overriding restriction: the frequency of polygon types correlates with the distribution of cell areas. By altering the balance of tensions and pressures within the packed tissues using disease, genetic or computer model perturbations, we show that as long as packed cells present a balance of forces within tissue, they will be under a physical constraint that limits its organization. Our discoveries establish a new framework to understand tissue architecture in development and disease.