Molecular docking and 3D-quantitative structure activity relationship analyses of peptidyl vinyl sulfones: Plasmodium Falciparum cysteine proteases inhibitors

Comparative molecular field analysis (CoMFA) and comparative molecular similarity indices analysis (CoMSIA) based on three-dimensional quantitative structure- activity relationship (3D-QSAR) studies were conducted on a series (39 molecules) of peptidyl vinyl sulfone derivatives as potential Plasmodium Falciparum cysteine proteases inhibitors. Two different methods of alignment were employed: (i) a receptor-docked alignment derived from the structure-based docking algorithm GOLD and (ii) a ligand-based alignment using the structure of one of the ligands derived from a crystal structure from the PDB databank. The best predictions were obtained for the receptor-docked alignment with a CoMFA standard model (q(2) = 0.696 and r(2) = 0.980) and with CoMSIA combined electrostatic, and hydrophobic fields (q(2) = 0.711 and r(2) = 0.992). Both models were validated by a test set of nine compounds and gave satisfactory predictive r(2) pred values of 0.76 and 0.74, respectively. CoMFA and CoMSIA contour maps were used to identify critical regions where any change in the steric, electrostatic, and hydrophobic fields may affect the inhibitory activity, and to highlight the key structural features required for biological activity. Moreover, the results obtained from 3D-QSAR analyses were superimposed on the Plasmodium Falciparum cysteine proteases active site and the main interactions were studied. The present work provides extremely useful guidelines for future structural modifications of this class of compounds towards the development of superior antimalarials

Coxiella burnetii Infection in Livestock, Pets, Wildlife, and Ticks in Latin America and the Caribbean: a Comprehensive Review of the Literature

Purpose of the Review Q fever , a bacterial zoonosis caused by Coxiella burnetii, is reported very heterogeneously in humans in Latin America. The objective of this study was to review the data on Coxiella burnetii Infection in animals in Latin America and the Caribbean. Recent Findings A comprehensive literature review was carried out in the 47 countries and territories of Latin America on various search engines and grouped into four groups: livestock, pets, wildlife, and ticks. Summary Thus, 113 studies were selected between 1950 and 2022. Among the 47 countries, only 25 (53%) had at least one publication related to C. burnetii infection in animals. The most productive country was Brazil (N = 51), followed by French Guiana (N = 21), and Colombia (N = 16). Studies in livestock from 20 countries have shown widely varying country-to-country rates of seroprevalence, ranging from 0 to 67%. Some studies from seven countries, especially French Guiana and Brazil, found antibodies and sometimes positive PCR in dogs and cats, generally in the context of investigations around human clustered cases. Knowledge remained fragmented about infection in wildlife from only five countries (Chile, Colombia, Brazil, French Guiana, and Uruguay). C. burnetii infection was identified by PCR in Chiroptera (7 species), Rodentia (6 species), Suina (2 species), Xenartha (1 species), Cingulata (1 species), and Perissodactyla (1 species). Studies on Coxiella sp. in ticks have been performed in 11 countries, mostly in Brazil, and mainly found Coxiella-like endosymbionts. Thus, data on C. burnetii infection in animals are sparse and incomplete in Latin America and the Caribbean, and more research is warranted

Preferential Entry of Botulinum Neurotoxin A Hc Domain through Intestinal Crypt Cells and Targeting to Cholinergic Neurons of the Mouse Intestine

Botulism, characterized by flaccid paralysis, commonly results from botulinum neurotoxin (BoNT) absorption across the epithelial barrier from the digestive tract and then dissemination through the blood circulation to target autonomic and motor nerve terminals. The trafficking pathway of BoNT/A passage through the intestinal barrier is not yet fully understood. We report that intralumenal administration of purified BoNT/A into mouse ileum segment impaired spontaneous muscle contractions and abolished the smooth muscle contractions evoked by electric field stimulation. Entry of BoNT/A into the mouse upper small intestine was monitored with fluorescent HcA (half C-terminal domain of heavy chain) which interacts with cell surface receptor(s). We show that HcA preferentially recognizes a subset of neuroendocrine intestinal crypt cells, which probably represent the entry site of the toxin through the intestinal barrier, then targets specific neurons in the submucosa and later (90–120 min) in the musculosa. HcA mainly binds to certain cholinergic neurons of both submucosal and myenteric plexuses, but also recognizes, although to a lower extent, other neuronal cells including glutamatergic and serotoninergic neurons in the submucosa. Intestinal cholinergic neuron targeting by HcA could account for the inhibition of intestinal peristaltism and secretion observed in botulism, but the consequences of the targeting to non-cholinergic neurons remains to be determined

Botulinum hemagglutinin disrupts the intercellular epithelial barrier by directly binding E-cadherin

Botulinum neurotoxin's nontoxic HA protein binds E-cadherin to disrupt cell–cell adhesion in a species-specific manner

Striking HIV-1 Entry by Targeting HIV-1 gp41. But, Where Should We Target?

HIV-1 gp41 facilitates the viral fusion through a conformational switch involving the association of three C-terminal helices along the conserved hydrophobic grooves of three N-terminal helices coiled-coil. The control of these structural rearrangements is thought to be central to HIV-1 entry and, therefore, different strategies of intervention are being developed. Herewith, we describe a procedure to simulate the folding of an HIV-1 gp41 simplified model. This procedure is based on the construction of plausible conformational pathways, which describe protein transition between non-fusogenic and fusogenic conformations. The calculation of the paths started with 100 molecular dynamics simulations of the non-fusogenic conformation, which were found to converge to different intermediate states. Those presenting defined criteria were selected for separate targeted molecular dynamics simulations, subjected to a force constant imposing a movement towards the gp41 fusogenic conformation. Despite significant diversity, a preferred sequence of events emerged when the simulations were analyzed in terms of the formation, breakage and evolution of the contacts. We pointed out 29 residues as the most relevant for the movement of gp41; also, 2696 possible interactions were reduced to only 48 major interactions, which reveals the efficiency of the method. The analysis of the evolution of the main interactions lead to the detection of four main behaviors for those contacts: stable, increasing, decreasing and repulsive interactions. Altogether, these results suggest a specific small cavity of the HIV-1 gp41 hydrophobic groove as the preferred target to small molecules

Bacterial Toxins and the Nervous System: Neurotoxins and Multipotential Toxins Interacting with Neuronal Cells

Toxins are potent molecules used by various bacteria to interact with a host organism. Some of them specifically act on neuronal cells (clostridial neurotoxins) leading to characteristics neurological affections. But many other toxins are multifunctional and recognize a wider range of cell types including neuronal cells. Various enterotoxins interact with the enteric nervous system, for example by stimulating afferent neurons or inducing neurotransmitter release from enterochromaffin cells which result either in vomiting, in amplification of the diarrhea, or in intestinal inflammation process. Other toxins can pass the blood brain barrier and directly act on specific neurons

Endomicroscopy and electromyography of neuromuscular junctions in situ

OBJECTIVE: Electromyography (EMG) is used routinely to diagnose neuromuscular dysfunction in a wide range of peripheral neuropathies, myopathies, and neuromuscular degenerative diseases including motor neuron diseases such as amyotrophic lateral sclerosis (ALS). Definitive neurological diagnosis may also be indicated by the analysis of pathological neuromuscular innervation in motor-point biopsies. Our objective in this study was to preempt motor-point biopsy by combining live imaging with electrophysiological analysis of slow degeneration of neuromuscular junctions (NMJs) in vivo. METHODS: We combined conventional needle electromyography with fiber-optic confocal endomicroscopy (CEM), using an integrated hand-held, 1.5-mm-diameter probe. We utilized as a test bed, various axotomized muscles in the hind limbs of anaesthetized, double-homozygous thy1.2YFP16: Wld(S) mice, which coexpress the Wallerian-degeneration Slow (Wld(S)) protein and yellow fluorescent protein (YFP) in motor neurons. We also tested exogenous vital stains, including Alexa488-α-bungarotoxin; the styryl pyridinium dye 4-Di-2-Asp; and a GFP conjugate of botulinum toxin Type A heavy chain (GFP-HcBoNT/A). RESULTS: We show that an integrated EMG/CEM probe is effective in longitudinal evaluation of functional and morphological changes that take place over a 7-day period during axotomy-induced, slow neuromuscular synaptic degeneration. EMG amplitude declined in parallel with overt degeneration of motor nerve terminals. EMG/CEM was safe and effective when nerve terminals and motor endplates were selectively stained with vital dyes. INTERPRETATION: Our findings constitute proof-of-concept, based on live imaging in an animal model, that combining EMG/CEM may be useful as a minimally invasive precursor or alternative to motor-point biopsy in neurological diagnosis and for monitoring local administration of potential therapeutics

Passage de la neurotoxine botulique à travers la barrière intestinale

paralysis, is produced by anaerobic bacteria of the Clostridium genus. BoNTs are classified into 7 types (A to G) and form different complexes by association with non-toxic proteins, including the non-toxic non-hemagglutinin component (NTNH) and hemagglutinins (HAs). En coding genes are clustered in the botulinum locus and organized in two divergent operons, ntnh-bont/A and ha34-ha17-ha70 for type A, whose expressions are positively regulated by the alternative sigma factor BotR. A synchronous expression peak of all the botulinum locus genes is measured by real-time RT-PCR at the transition between exponential and stationary growth phases, concomitantly to toxin accumulation in culture supernatant. In an intestinal epithelium model, purified BoNT/A is transcytosed after binding, via the Hc/A domain, to apical receptors including gangliosides and putative SV2-related proteins. Binding affinity and transport efficiency into intestinal cells are higher with crypt-type than with enterocytetype cell lines. Injected into the lumen of ligatured mouse ileal loops, BoNT/A inhibits smooth muscle contractions and fluorescent Hc/A migrates across mucosa, through some crypt cells, to submucosa and musculosa where it targets certain nerve endings, mostly cholinergic. Hc /A is internalized via different pathways into neuronal cells (dynamin- and clathrin-dependent) and intestinal cells (dynamin- and Cdc42-dependent, and clathrinindependent).flasque, est produite par des bactéries anaérobies du genre Clostridium. Les BoNTs, classifiées en 7 types (A à G), forment divers complexes avec des protéines non toxiques, dont le composant non toxique non hémagglutinant (NTNH) et les hémagglutinines (HAs). Les gènes sont organisés, au sein du locus botulique, selon deux opérons divergents, ntnhbont/ A et ha34-ha17-ha70 pour le type A, dont l'expression est positivement régulée par le facteur sigma alternatif BotR. Un pic d'expression synchrone de tous les gènes du locus de type A est mesuré par RT-PCR en temps réel lors de la transition entre les phases exponentielle et stationnaire de croissance, en parallèle avec l'augmentation du titre en toxine du surnageant de culture. Dans un modèle d'épithélium intestinal, BoNT/A purifiée est transcytosée après liaison via le domaine Hc/A à des récepteurs apicaux comprenant des gangliosides et des protéines potentiellement apparentées à SV2. L'intensité de liaison et l'efficacité de transport de la toxine sont supérieures dans les cellules intestinales de type crypte plutôt qu'entérocyte. Injectés dans la lumière d'une anse iléale ligaturée, BoNT/A inhibe les contractions des muscles lisses et le domaine Hc/A fluorescent progresse de la muqueuse, via certaines cellules des cryptes, vers la sous-muqueuse et la musculeuse où il cible certaines terminaisons nerveuses, majoritairement cholinergiques. Hc/A entre par des voies distinctes dans les cellules neuronales (voie clathrine dépendante de la dynamine) et les cellules intestinales (voie non-clathrine, dépendante de Cdc42 et de la dynamine)