Inhibition of a malaria host–pathogen interaction by a computationally designed inhibitor

Malaria is a substantial global health burden with 229 million cases in 2019 and 450,000 deaths annually. Plasmodium vivax is the most widespread malaria-causing parasite putting 2.5 billion people at risk of infection. P. vivax has a dormant liver stage and therefore can exist for long periods undetected. Its blood-stage can cause severe reactions and hospitalization. Few treatment and detection options are available for this pathogen. A unique characteristic of P. vivax is that it depends on the Duffy antigen/receptor for chemokines (DARC) on the surface of host red blood cells for invasion. P. vivax employs the Duffy binding protein (DBP) to bind to DARC. We first de novo designed a three helical bundle scaffolding database which was screened via protease digestions for stability. Protease-resistant scaffolds highlighted thresholds for stability, which we utilized for selecting DARC mimetics that we subsequentially designed through grafting and redesign of these scaffolds. The optimized design small helical protein disrupts the DBP:DARC interaction. The inhibitor blocks the receptor binding site on DBP and thus forms a strong foundation for a therapeutic that will inhibit reticulocyte infection and prevent the pathogenesis of P. vivax malaria.     

The levator ani of the female rat: a suitable model for studying the effects of testosterone on the development of mammalian muscles

The present results give evidence by using cytochemical markers of motor end-plates (ChE, AchR) and ultrastructural techniques that the Levator ani (LA) muscle is also present in adult females: it is composed of differentiated and innervated fibres. A significant difference, both on the number of fibres per muscle (n) and on their average cross-sectional area (acsa), was observed between sexes:-male: n = 5300 +/- 687, acsa: 522 +/- 68.6 microns 2;-female: n = 565 +/- 246.9, acsa: 68 +/- 8.6 microns 2. These results suggest that testosterone could control, at least partially, the number and the diameter of muscle fibres during development.     

Implications of Ordovician (≈460 Myr) marine cement for constraining seawater temperature and atmospheric pCO2

The marine diagenetic history of the Effna buildups (Virginia, lower Caradocian, ≈460 Myr) is atypical of Phanerozoic carbonates and reflects variable redox conditions in the semi-restricted Appalachian foreland basin. Fibrous marine cements are the focus of this study. Least-altered fibrous calcite from the Effna Formation has a translucent appearance in transmitted light and a 2-3 mol% MgCO₃ composition; the latter is similar to values (2-5 mol%) from translucent fibrous calcite in other Caradocian units [Holston (Tennessee) and Kullsberg (Sweden) formations]. Some fibrous calcite is overlain by hardgrounds that in places have an irregular micro-topography with sharp overhangs indicative of syn-depositional dissolution. Geochemical evidence supporting the marine dissolution includes δ¹⁸O values from altered fibrous calcite that are more positive (up to −3.9‰ PDB) than coeval least-altered fibrous calcite from Holston buildups in Tennessee (up to −4.8 to −5.8‰ PDB). Syn-depositional dissolution was likely associated with sulfide oxidation along an oxic-anoxic interface and not due to upwelling of cold bottom water in the Appalachian foredeep. Faunal and lithologic evidence suggests that oxic surface waters had a normal salinity and a water temperature conducive for metazoan growth (<33°C). This paper constrains the temperature of near-surface seawater in the Appalachian foreland basin (22-33°C) and atmospheric pCO₂ (>14 present atmospheric level) during the early Caradocian.     

Split versions of Cleave and Rescue selfish genetic elements for measured self limiting gene drive

Gene drive elements promote the spread of linked traits, providing methods for changing the composition or fate of wild populations. Drive mechanisms that are self-limiting are attractive because they allow control over the duration and extent of trait spread in time and space, and are reversible through natural selection as drive wanes. Self-sustaining Cleave and Rescue (ClvR) elements include a DNA sequence-modifying enzyme such as Cas9/gRNAs that disrupts endogenous versions of an essential gene, a tightly linked recoded version of the essential gene resistant to cleavage (the Rescue), and a Cargo. ClvR spreads by creating loss-of-function (LOF) conditions in which those without ClvR die because they lack functional copies of the essential gene. We use modeling to show that when the Rescue-Cargo and one or both components required for LOF allele creation (Cas9 and gRNA) reside at different locations (split ClvR), drive of Rescue-Cargo is self-limiting due to a progressive decrease in Cas9 frequency, and thus opportunities for creation of LOF alleles, as spread occurs. Importantly, drive strength and duration can be extended in a measured manner—which is still self-limiting—by moving the two components close enough to each other that they experience some degree of linkage. With linkage, Cas9 transiently experiences drive by hitchhiking with Rescue-Cargo until linkage disequilibrium between the two disappears, a function of recombination frequency and number of generations, creating a novel point of control. We implement split ClvR in Drosophila, with key elements on different chromosomes. Cargo/Rescue/gRNAs spreads to high frequency in a Cas9-dependent manner, while the frequency of Cas9 decreases. These observations show that measured, transient drive, coupled with a loss of future drive potential, can be achieved using the simple toolkit that make up ClvR elements—Cas9 and gRNAs and a Rescue/Cargo.     

Parasympathetic NANC-secretion of saliva in the mink, and effects of substance P and VIP.

In both parotid and submandibular glands a parasympathetic non-adrenergic, non-cholinergic (NANC) nerve-evoked secretion of saliva was demonstrated. Saliva evoked by exogenous substance P was poor in protein, while saliva evoked by VIP was protein-rich. In a subthreshold dose for fluid secretion VIP released protein and potentiated the responses elicited by substance P, particularly regarding the output of protein. The two neuropeptides may contribute to the parasympathetic NANC secretion of saliva in the mink. Further, agonists responsible for the secretory NANC response are also likely to contribute to the secretory response of the glands to parasympathetic stimulation in the absence of autonomic receptor blockade in this species.     

Optical rotation of some α-1,4-linked glucopyranosides in the system H2O–DMSO and solution conformation of amylose

The specific rotation of starch components, corrected for refractive index variation, exhibits a discontinuity in the region of the water–dimethyl sulfoxide (H₂O–DMSO) system that corresponds to the composition of the complex 2H₂O–DMSO. This discontinuity is a property dependent upon the presence of a number of consecutively linked α-1,4 glucose units and, therefore, must reflect a change in symmetry of a segment, of polymer chain. The optical rotation of amylose between 26.5 and 92.5°C. does not change in DMSO and is only slightly lowered in water at the higher temperature. The behavior of amylose in both DMSO and H₂O is like that of a random coil, as indicated by viscosity and sedimentation measurements. These results may be interpreted either as being compatible with models of amylose in solution in which the polymer backbone has helical twist, or as indicating removal of strong interactions between polymer chain segments by a good solvent.