Genome-wide CRISPR Screens Reveal Host Factors Critical for SARS-CoV-2 Infection

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Obesity-inducing lesions of the central nervous system alter leptin uptake by the blood-brain barrier.

Leptin regulates body adiposity by decreasing feeding and increasing thermogenesis. Obese humans and some obese rodents are resistant to peripherally administered leptin, suggesting a defect in the transport of leptin across the blood-brain barrier (BBB). Defective transport of exogenous leptin occurs in some models of obesity, but in other models transport is normal. This shows that factors other than obesity are associated with impairment of leptin transport across the BBB. In order to further investigate these factors, we determined leptin transport in rats made obese by lesioning of the ventromedial hypothalamus (VMH), paraventricular nucleus (PVN), or posterodorsal amygdala (PDA). These regions all contain leptin receptors and lesions there induce obesity and hyperleptinemia and alter the levels of many feeding hormones which might participate in leptin transporter regulation. We measured the uptake of radioactively labeled leptin by the BBB by multiple-time regression analysis which divides uptake into a reversible phase (Vi, e.g., receptor/transporter binding to the brain endothelial cell) and an irreversible phase (Ki, complete transport across the BBB). Leptin uptake was not affected in rats with VMH lesions. No significant change occurred in the entry rate (Ki) for any group, although Ki declined by over 35% in rats with PVN lesions. Decreased uptake was observed in rats with PVN lesions and with PDA lesions. This was primarily due to a reduced Vi (about 21% for the PDA). This decreased uptake is most likely explained by decreased binding of leptin to the brain endothelial cell, which could be because of decreased binding by either receptors or transporters. This suggests that some of the feeding hormones controlled by the PVN and PDA may participate in regulating leptin uptake by the BBB.     

Vasculature of the head and respiratory organs in an obligate air-breathing fish,the swamp eel Monopterus (=Amphipnous) cuchia

Methyl methacrylate vascular corrosion replicas were used to examine the macrocirculation in the head region and the microcirculation of respiratory vessels in the air-breathing swamp eel Monopterus cuchia. Fixed respiratory tissue was also examined by SEM to verify capillary orientation. The respiratory and systemic circulations are only partially separated, presumably resulting in supply of mixed oxygenated and venous blood to the tissues. A long ventral aorta gives rise directly to the coronary and hypobranchial arteries. Two large shunt vessels connect the ventral aorta to the dorsal aorta, whereas the remaining ventral aortic flow goes to the respiratory islets and gills. Only two pairs of vestigial gill arches remain, equivalent to the second and third arches, yet five pairs of aortic arches were identified. Most aortic arches supply the respiratory islets. Respiratory islet capillaries are tightly coiled spirals with only a fraction of their total length in contact with the respiratory epithelium. Valve-like endothelial cells delimit the capillary spirals and are unlike endothelial cells in other vertebrates. The gills are highly modified in that the lamellae are reduced to a single-channel capillary with a characteristic three-dimensional zig-zag pathway. There are no arterio-arterial lamellar shunts, although the afferent branchial artery supplying the gill arches also supplies respiratory islets distally. A modified interlamellar filamental vasculature is present in gill tissue but absent or greatly reduced in the respiratory islets. The macro- and micro-circulatory systems of M. cuchia have been considerably modified presumably to accommodate aerial respiration. Some of these modifications involve retention of primitive vessel types, whereas others, especially in the microcirculation, incorporate new architectural designs some of whose functions are not readily apparent.     

Foraging Efficacy of a Larval Parasitoid in a Cotton Patch: Influence of Chemical Cues and Learning

Plant-herbivore chemical signals and behavioral plasticity may enhance parasitoid host-foraging efficacy in the field; however, no studies have quantified the potential benefits from these factors under field-type conditions. The effect of plant-herbivore signals and learning on the foraging efficacy of Microplitis croceipes was quantified by directly observing and recording total and sequential duration of various foraging behaviors relative to 5 randomly placed herbivore-damaged and host-infested cotton plants and 20 undamaged and non-host-infested plants. Microplitis croceipes spent significantly more time searching (flying and antennation) on host infested versus uninfested plants. Antennation time was significantly and negatively correlated with successive host stings. Contrary to expectations of increased duration, flight time remained constant throughout the foraging bout, which may indicate that there was some learning associated with flight. These results suggest that plant-herbivore chemical signals and learning enhances the foraging efficacy of M. croceipes.     

ADAPTATIONS TO CLIFF-NESTING IN SOME ARCTIC GULLS (LARUS)

Various aspects of the breeding of three gulls, Larus thayeri, glaucoides and hyperboreus, which nested in colonies both on cliff ledges and on level ground in the eastern Canadian Arctic, were compared with those of the ground-nesting L. argentatus and the cliff-nesting Rissa tridactyla. As the result of adaptation to cliff ledge nesting, many aspects of the breeding biology of R. tridactyla were strikingly different from those of the ground-nesting European L. argentatus, but the behaviour of L. thayeri, glaucoides and hyperboreus clearly spanned these differences. Cliff-nesting individuals of thayeri and glaucoides were most like Rissa; ground-nesting individuals of these species were most like argentatus. L. thayeri was more like Rissa than was glaucoides. With but few exceptions, both cliff- and ground-nesting individuals of hyperboreus were most like argentatus. The factors responsible for the intra-specific differences between cliff- and ground-nesters of thayeri and glaucoides are not clear. Limited gene exchange between cliff and ground colonies occurs. Because of physical features of the nest, first-laid eggs were more liable to fall from ledges than second or third eggs. L. thayeri and glaucoides have evolved separate mechanisms to cope with this problem. Egg shape was multimodal in thayeri and glaucoides. Long pyriform eggs were less liable to fall from ledges than eggs of other shapes. L. thayeri laid more long pyriform eggs as first eggs than did glaucoides. L. thayeri lost fewer eggs than did glaucoides, but glaucoides replaced all lost eggs while thayeri did not. Delayed follicular atresia provided glaucoides with insurance of egg replacement. In thayeri, accessory follicles were reabsorbed after the first egg was laid; in argentatus, after the second egg, and in glaucoides after the third egg. At the approach of a predator, it was advantageous for cliff-dwelling chicks to remain motionless but for ground-dwelling chicks to flee their nests and to hide. Among the cliff-nesters, the “freezing in place” reaction of chicks was best developed in thayeri, to a lesser extent in glaucoides, and least in hyperboreus. Among the ground-nesters, chicks of glaucoides and hyperboreus behaved like those of argentatus and fled their nests when disturbed, but chicks of thayeri froze like their cliff-dwelling siblings. Reciprocal transfers of eggs and chicks between cliff and ground colonies indicated that in argentatus, glaucoides and hyperboreus, the factors determining a chick's reaction to disturbance came into play between hatching and the eighth day. In thayeri, the reaction appeared to be effectively innate. Chicks of glaucoides showed a greater predisposition to this behaviour than chicks of argentatus after both had received identical experience on cliff ledges. In thayeri, stereotypy of the freezing reaction has probably been a factor limiting the colonisation of areas where cliffs are scarce but predators present. In argentatus, lack of perfection of this behaviour (compared to glaucoides and thayeri) has probably been a factor preventing argentatus from attaining cliff ledges. L. hyperboreus, although nowhere abundant, is a widespread species nesting on level ground and on cliff ledges but lacking the modifications observed in glaucoides and thayeri; this is due to its size and aggressiveness, the fact that it picks nest-sites before glaucoides and thayeri arrive in the colonies, and that on cliffs it chooses the largest and most level ledges.     

Ultrastructure of Volvox carteri

Summary Somatic cells of mature asexual colonies of Volvox carteri do not possess a true cell wall, but are otherwise similar in ultrastructure to Chlamydomonas. Somatic cells are embedded in multilayered fibrillar material of the colonial matrix. The reproductive cells (gonidia) of Volvox carteri lie internal to the somatic cell layer of the colony matrix in an apparently structureless portion of the colony matrix. Mature gonidia are large vacuolate cells with a central nucleus and parietal chloroplasts and mitochondria. They are non-flagellated at maturity, but each contains a pair of kinetosomes.     

Arginine utilization by Mycoplasma fermentans is not regulated by glucose metabolism: A 13C-NMR study

Abstract Electrofusion of protoplasts of two mutant strains of Hansenula polymorpha resulted in high fusion and hybrid yields when the calcium ions present in the conventional fusion medium replaced by zinc ions. The optimal fusion conditions were an alignment field of 0.4 kV cm⁻¹ strength and 2 MHz frequency for 30 s, followed by two consecutive pulses of 12 kV cm⁻¹ strength and 15 μs duration. With 0.05–0.1 mM zinc ions in the fusion medium an average clone number of 10⁴–10⁵ clones per 10⁸ input cells was reached. The presence of about 0.6 mM magnesium ions in the zinc fusion medium was essential.     

DNA sequence homology between attB-related sites of Corynebacterium diphtheriae, Corynebacterium ulcerans, Corynebacterium glutamicum, and the attP side of γ-Cornephage

Chromosomal restriction fragments of Corynebacterium ulcerans and C. diphtheriae, containing an integration site for corynephages of the beta family, show homology on Southern blots. Homologous DNA in also found in the soil isolate C. glutamicum, although this strain is not susceptible to beta-corynephages. Three of these DNA fragments, one for each bacterial strain, and a fragment of gamma-corynephage DNA previously shown to contain the phage integration site, were cloned and sequenced. Alignment of the 3 bacterial sequences shows a very high degree of homology in a stretch of ca 120 nucleotides, whereas the rest of the sequences is generally non-homologous. Within this common bacterial portion, a segment of ca. 96 nucleotides (core sequence) is also highly homologous to the phage sequence. The first half (ca. 50 bp) of the core sequence is identical in all aligned sequences whereas the second half, which is largely occupied by a stem-and-loop structure, contains point mutations peculiar to each clone. The described sequences are likely to be involved in phage integration/excision processes.