Interactions between Allosteric Modulators and 4-DAMP and Other Antagonists at Muscarinic Receptors: Potential Significance of the Distance between the N and Carboxyl C Atoms in the Molecules of Antagonists

Allosteric enhancement of the affinity of muscarinic receptors for their ligands offers a new way to influence cholinergic neurotransmission. The structure of the allosteric binding domain(s) and the features of agonists, antagonists and modulators which determine the occurrence of either positive or negative cooperativity require clarification. We tested interactions between allosteric modulators alcuronium, strychnine and brucine and eight antagonists at muscarinic receptors expressed in CHO cells. In experiments with unlabeled antagonists, all three modulators enhanced the affinity for 4-diphenylacetoxy-N-dimethylpiperidinium (4-DAMP) at the M₂ receptors, and strychnine did so also at the M₄ receptors. Positive interactions were also observed between alcuronium and L-hyoscyamine (M₂) and scopolamine (M₂), between strychnine and butylscopolamine (M₄), L-hyoscyamine (M₂ and M₄) and scopolamine (M₄), and between brucine and scopolamine (M₂). Positive effects of alcuronium, strychnine and brucine on the affinity of the M₂ receptors for 4-DAMP have been confirmed by direct measurements of the binding of [³H]-4-DAMP. A comparison of molecular models of several antagonists which are esters revealed that antagonists in which the distance between the N and the carboxyl C atoms corresponds to five chemical bonds are more likely to display positive cooperativity with alcuronium at the M₂ receptors than the antagonists in which the N-carboxyl C distance corresponds to four chemical bonds.     

Effect of suramin on the human pathogen Candida albicans: implications on the fungal development and virulence

Candida albicans is an opportunistic pathogen that is of growing medical importance because it causes superficial, mucosal and systemic infections in susceptible individuals. Here, the effect of suramin, a polysulfonated naphthylurea derivative, on C. albicans development and virulence was evaluated. Firstly, it was demonstrated that suramin (500 microM) arrested its growth, showing a fungicidal action dependent on cell number. Suramin treatment caused profound changes in the yeast ultrastructure as shown by transmission electron microscopy. The more important changes were the enlargement of the fungi cytoplasmic vacuoles, the appearance of yeasts with an empty cytoplasm resembling ghost cells and a reduction in cell wall thickness. Suramin also blocked the transformation of yeast cells to the germ-tube and the interaction between C. albicans and epithelial cells. In order to ascertain that the action of suramin on C. albicans growth is a general feature instead of being strain-specific, the effects of suramin on 14 oral clinical strains isolated from healthy children and HIV-positive infants were analyzed. Interestingly, the strains of C. albicans isolated from HIV-positive patients were more resistant to suramin than strains isolated from healthy patients. Altogether, the results produced here show that suramin interfered with essential fungal processes, such as growth, differentiation and interaction with host cells.     

Compartment‐specific aggregases direct distinct nuclear and cytoplasmic aggregate deposition

Disruption of the functional protein balance in living cells activates protective quality control systems to repair damaged proteins or sequester potentially cytotoxic misfolded proteins into aggregates. The established model based on Saccharomyces cerevisiae indicates that aggregating proteins in the cytosol of eukaryotic cells partition between cytosolic juxtanuclear (JUNQ) and peripheral deposits. Substrate ubiquitination acts as the sorting principle determining JUNQ deposition and subsequent degradation. Here, we show that JUNQ unexpectedly resides inside the nucleus, defining a new intranuclear quality control compartment, INQ, for the deposition of both nuclear and cytosolic misfolded proteins, irrespective of ubiquitination. Deposition of misfolded cytosolic proteins at INQ involves chaperone‐assisted nuclear import via nuclear pores. The compartment‐specific aggregases, Btn2 (nuclear) and Hsp42 (cytosolic), direct protein deposition to nuclear INQ and cytosolic (CytoQ) sites, respectively. Intriguingly, Btn2 is transiently induced by both protein folding stress and DNA replication stress, with DNA surveillance proteins accumulating at INQ. Our data therefore reveal a bipartite, inter‐compartmental protein quality control system linked to DNA surveillance via INQ and Btn2.     

Cellular characterisation of Candida tropicalis presenting fluconazole-related trailing growth

We assessed fluconazole susceptibility in 52 Candida tropicalis clinical strains using seven antifungal susceptibility methods, including broth microdilution (BMD) [standard M27 A3 (with neutral and acid pH), ATB Fungus 3, Vitek 2 system and flow cytometric analysis] and agar-based methods (disk diffusion and E-test). Trailing growth, detection of cell-associated secreted aspartic proteases (Saps) and morphological and ultrastructural traits of these clinical strains were also examined. The ranges of fluconazole 24 h-minimum inhibitory concentration (MIC) values were similar among all methods. The essential agreement among the methods used for MIC determinations was excellent and all methods categorised all strains as susceptible, except for one strain that showed a minor error. The presence of the trailing effect was assessed by six methods. Trailing positivity was observed for 86.5-100% of the strains. The exception was the BMD-Ac method where trailing growth was not observed. Morphological and ultrastructural alterations were detected in C. tropicalis trailing cells, including mitochondrial swelling and cell walls with irregular shapes. We tested the production of Saps in 13 C. tropicalis strains expressing trailing growth through flow cytometry. Our results showed that all of the C. tropicalis strains up-regulated surface Sap expression after 24 h or 48 h of exposure to fluconazole, which was not observed in untreated yeast strains. We concluded that C. tropicalis strains expressing trailing growth presented some particular features on both biological and ultrastructural levels.     

Towards resolving conservation issues through historical aerial imagery: vegetation cover changes in the Central European tundra

Mountain ecosystems are considered to be sensitive to global change and human disturbance. Our retrospective analysis of archival aerial imagery showed dynamics of arctic-alpine tundra vegetation in the Krkono?e Mountains, Czech Republic; image classification revealed a change in land cover classes in 44% of the study area over the past eighty years. This particular ecosystem holds many rare features, such as high numbers of endemic, glacial relict and rare species as well as relict geomorphological components such as sorted patterned ground. Our study revealed an accelerating expansion of the native and planted shrub, Pinus mugo, from 30.6% of the study area in 1936 to 48.6% in 2018, mostly at the expense of grasslands that decreased from 59.3% in 1936 to 44.2% in 2018. Shrub expansion represents a threat to relict periglacial landforms, covering 8% of the sorted patterned ground in 1936 and 26.5% in 2018. Shrub encroachment was shown to be due to artificial planting of the pine in the past as well as the cessation of former farming (mowing and cattle grazing) and, most probably, also by global change. Both dwarf pine stands and tundra grasslands hold high conservation value (Natura 2000 habitats); a balance between different nature protection interests must, therefore, be ensured. Detailed spatio-temporally, explicit outputs of the remote sensing analysis can serve as a baseline for nature conservation in order to prepare corresponding management plans.

     

Rethinking Human–Nonhuman Primate Contact and Pathogenic Disease Spillover

Zoonotic transmissions are a major global health risk, and human–animal contact is frequently raised as an important driver of transmission. A literature examining zooanthroponosis largely agrees that more human–animal contact leads to more risk. Yet the basis of this proposition, the term contact, has not been rigorously analyzed. To understand how contact is used to explain cross-species spillovers, we conducted a multi-disciplinary review of studies addressing human–nonhuman primate (NHP) engagements and pathogenic transmissions and employing the term contact. We find that although contact is frequently invoked, it is employed inconsistently and imprecisely across these studies, overlooking the range of pathogens and their transmission routes and directions. We also examine a related but more expansive approach focusing on human and NHP habitats and their spatial overlap, which can potentially facilitate pathogenic transmission. Contact and spatial overlap investigations cannot, however, explain the processes that bring together people, animals and pathogens. We therefore examine another approach that enhances our understanding of zoonotic spillovers: anthropological studies identifying such historical, social, environmental processes. Comparable to a One Health approach, our ongoing research in Cameroon draws contact, spatial overlap and anthropological–historical approaches into dialog to suggest where, when and how pathogenic transmissions between people and NHPs may occur. In conclusion, we call for zoonotic disease researchers to specify more precisely the human–animal contacts they investigate and to attend to how broader ecologies, societies and histories shape pathogen–human–animal interactions.     

Impact of Two Accidentally Introduced Encarsia Species (Hymenoptera: Aphelinidae) and Other Biotic and Abiotic Factors on the Spiralling Whitefly Aleurodicus dispersus (Russell) (Homoptera: Aleyrodidae), in Benin, West Africa

In early 1993, the spiralling whitefly, Aleurodicus dispersus (Russell), was observed in Benin for the first time, inflicting damage to ornamental and shade trees and cassava. The parasitoids Encarsia ?haitiensis Dozier and E. guadeloupae Viggiani were observed in the second half of 1993. They were known to have the same host in the Pacific region, and were thought to have been introduced accidentally. The impact of these parasitoids was quantified using four surveys from 1993 to 1995 (on 2541 trees in 537 localities) and by population studies on guava. In 1993, A. dispersus occurred mostly in towns in the southern part of Benin; penetration into farmland was observed later. E. ?haitiensis was more abundant and widespread than E. guadeloupae , and by 1995 it had been recovered from most (84%) of the infested localities. On guava trees, the annual peaks of A. dispersus population declined by ca. 80% between 1993 and 1996. During the same period parasitism rates increased. Econometric multiple regression analyses based on 996 infested trees demonstrated that A. dispersus population densities, the proportion of infested trees and damage scores all declined significantly with increasing duration of the presence of the parasitoids, indicating their impact. Other variables were also significantly related to A. dispersus levels.