Antagonists with a long residence time at the receptors are desired for the possibility of reducing daily doses and side effects. KH-5 (1-{2-[4-(hexyloxy)benzoyloxy]ethyl}-1-methyl-1,2,3,6-tetrahydropyridin-1-ium iodide) is the long-acting M1-preferring bitopic muscarinic antagonist with a half-life at muscarinic receptors of up to five hours. The binding of 2-hexyloxy and 3-hexyloxy analogues of KH-5 was simulated in silico, compounds were synthesized and their binding and antagonistic properties were measured experimentally in CHO cells expressing individual subtypes of muscarinic acetylcholine receptors. The overall binding affinities of the new compounds were similar to their respective parent compounds. Shifting the hexyloxy chain to ortho and meta positions led to a decrease in potency at the M1 receptor but an increase in potency at the M2 receptor and abolition of long-term antagonism. Preservation of the para position of the hexyloxy chain is essential for the further development of M1-preferring antagonists. Modifications of the basic centre may be the way to improve the geometry of antagonists towards long residence times to obtain the desired long-acting muscarinic antagonists in the future. The additional challenge for further development is the low metabolic stability of compounds.

• MeSH
• Muscarinic Antagonists * pharmacology   MeSH
• CHO Cells MeSH
• Cricetulus * MeSH
• Pyridines pharmacology   chemistry   MeSH
• Receptor, Muscarinic M1 metabolism   antagonists & inhibitors   MeSH
• Receptors, Muscarinic metabolism   MeSH
• Structure-Activity Relationship MeSH
• Animals MeSH
• Check Tag
• Animals MeSH
• Publication type
• Journal Article MeSH

Hyperthermia along with hydrocortisone (HC) are proven teratogens that can negatively influence embryo development during early pregnancy. Proliferation of cells is one of the main developmental processes during the early embryogenesis. This study was focused on testing the effect of elevated temperature and HC addition on proliferation of cells in in vitro cultures. The V79-4 cell line was treated with HC and cultured in vitro at 37 °C or 39 °C, respectively. To reveal the effect of both factors, the proliferation of cells cultured under different conditions was evaluated using various approaches (colony formation assay, generation of growth curves, computation of doubling times, and mitotic index estimation). Our results indicate that a short-term exposure to elevated temperature slightly stimulates and a long-term exposure suppresses cell proliferation. However, HC (0.1 mg/ml) acts as a stimulator of cell proliferation. Interestingly, the interaction of HC and long-term elevated temperature (39 °C) exposure results in at least partial compensation of the negative impact of elevated temperature by HC addition and in higher proliferation if compared with cells cultured at 39 °C without addition of HC.

• MeSH
• Cricetulus MeSH
• Fibroblasts * drug effects   cytology   metabolism   MeSH
• Hydrocortisone * pharmacology   MeSH
• Cells, Cultured MeSH
• Cell Proliferation * drug effects   MeSH
• Temperature MeSH
• Hot Temperature MeSH
• Animals MeSH
• Check Tag
• Animals MeSH
• Publication type
• Journal Article MeSH

Leishmania, the dixenous trypanosomatid parasites, are the causative agents of leishmaniasis currently divided into four subgenera: Leishmania, Viannia, Sauroleishmania, and the recently described Mundinia, consisting of six species distributed sporadically all over the world infecting humans and/or animals. These parasites infect various mammalian species and also cause serious human diseases, but their reservoirs are unknown. Thus, adequate laboratory models are needed to enable proper research of Mundinia parasites. In this complex study, we compared experimental infections of five Mundinia species (L. enriettii, L. macropodum, L. chancei, L. orientalis, and four strains of L. martiniquensis) in three rodent species: BALB/c mouse, Chinese hamster (Cricetulus griseus) and steppe lemming (Lagurus lagurus). Culture-derived parasites were inoculated intradermally into the ear pinnae and progress of infection was monitored for 20 weeks, when the tissues and organs of animals were screened for the presence and quantity of Leishmania. Xenodiagnoses with Phlebotomus duboscqi were performed at weeks 5, 10, 15 and 20 post-infection to test the infectiousness of the animals throughout the experiment. BALB/c mice showed no signs of infection and were not infectious to sand flies, while Chinese hamsters and steppe lemmings proved susceptible to all five species of Mundinia tested, showing a wide spectrum of disease signs ranging from asymptomatic to visceral. Mundinia induced significantly higher infection rates in steppe lemmings compared to Chinese hamsters, and consequently steppe lemmings were more infectious to sand flies: In all groups tested, they were infectious from the 5th to the 20th week post infection. In conclusion, we identified two rodent species, Chinese hamster (Cricetulus griseus) and steppe lemming (Lagurus lagurus), as candidates for laboratory models for Mundinia allowing detailed studies of these enigmatic parasites. Furthermore, the long-term survival of all Mundinia species in steppe lemmings and their infectiousness to vectors support the hypothesis that some rodents have the potential to serve as reservoir hosts for Mundinia.

• MeSH
• Arvicolinae * parasitology   MeSH
• Cricetulus MeSH
• Cricetinae MeSH
• Leishmania * classification   MeSH
• Leishmaniasis * parasitology   MeSH
• Disease Models, Animal * MeSH
• Mice, Inbred BALB C * MeSH
• Mice MeSH
• Animals MeSH
• Check Tag
• Cricetinae MeSH
• Mice MeSH
• Female MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH

Selective activation of individual subtypes of muscarinic receptors is a promising way to safely alleviate a wide range of pathological conditions in the central nervous system and the periphery as well. The flexible G-protein interface of muscarinic receptors allows them to interact with several G-proteins with various efficacy, potency, and kinetics. Agonists biased to the particular G-protein mediated pathway may result in selectivity among muscarinic subtypes and, due to the non-uniform expression of individual G-protein alpha subunits, possibly achieve tissue specificity. Here, we demonstrate that novel tetrahydropyridine-based agonists exert specific signalling profiles in coupling with individual G-protein α subunits. These signalling profiles profoundly differ from the reference agonist carbachol. Moreover, coupling with individual Gα induced by these novel agonists varies among subtypes of muscarinic receptors which may lead to subtype selectivity. Thus, the novel tetrahydropyridine-based agonist can contribute to the elucidation of the mechanism of pathway-specific activation of muscarinic receptors and serve as a starting point for the development of desired selective muscarinic agonists.

• MeSH
• Muscarinic Agonists * pharmacology   MeSH
• CHO Cells MeSH
• Cricetulus MeSH
• Carbachol pharmacology   MeSH
• Humans MeSH
• GTP-Binding Protein alpha Subunits metabolism   genetics   MeSH
• GTP-Binding Proteins metabolism   MeSH
• Pyridines pharmacology   MeSH
• Receptors, Muscarinic * metabolism   MeSH
• Signal Transduction drug effects   MeSH
• Animals MeSH
• Check Tag
• Humans MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH

The thermo- and pain-sensitive Transient Receptor Potential Melastatin 3 and 8 (TRPM3 and TRPM8) ion channels are functionally associated in the lipid rafts of the plasma membrane. We have already described that cholesterol and sphingomyelin depletion, or inhibition of sphingolipid biosynthesis decreased the TRPM8 but not the TRPM3 channel opening on cultured sensory neurons. We aimed to test the effects of lipid raft disruptors on channel activation on TRPM3- and TRPM8-expressing HEK293T cells in vitro, as well as their potential analgesic actions in TRPM3 and TRPM8 channel activation involving acute pain models in mice. CHO cell viability was examined after lipid raft disruptor treatments and their effects on channel activation on channel expressing HEK293T cells by measurement of cytoplasmic Ca2+ concentration were monitored. The effects of treatments were investigated in Pregnenolone-Sulphate-CIM-0216-evoked and icilin-induced acute nocifensive pain models in mice. Cholesterol depletion decreased CHO cell viability. Sphingomyelinase and methyl-beta-cyclodextrin reduced the duration of icilin-evoked nocifensive behavior, while lipid raft disruptors did not inhibit the activity of recombinant TRPM3 and TRPM8. We conclude that depletion of sphingomyelin or cholesterol from rafts can modulate the function of native TRPM8 receptors. Furthermore, sphingolipid cleavage provided superiority over cholesterol depletion, and this method can open novel possibilities in the management of different pain conditions.

• MeSH
• Analgesics pharmacology   therapeutic use   MeSH
• beta-Cyclodextrins * pharmacology   MeSH
• Pain chemically induced   drug therapy   metabolism   MeSH
• CHO Cells MeSH
• Cholesterol metabolism   MeSH
• Cricetulus MeSH
• HEK293 Cells MeSH
• TRPM Cation Channels * metabolism   genetics   MeSH
• Humans MeSH
• Membrane Microdomains metabolism   drug effects   MeSH
• Disease Models, Animal MeSH
• Mice MeSH
• Pregnenolone pharmacology   MeSH
• Pyrimidinones pharmacology   MeSH
• Sphingomyelin Phosphodiesterase * metabolism   pharmacology   MeSH
• Cell Survival drug effects   MeSH
• Animals MeSH
• Check Tag
• Humans MeSH
• Mice MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH

In late 2023, several SARS-CoV-2 XBB descendants, notably EG.5.1, were predominant worldwide. However, a distinct SARS-CoV-2 lineage, the BA.2.86 variant, also emerged. BA.2.86 is phylogenetically distinct from other Omicron sublineages, accumulating over 30 amino acid mutations in its spike protein. Here, we examined the virological characteristics of the BA.2.86 variant. Our epidemic dynamics modeling suggested that the relative reproduction number of BA.2.86 is significantly higher than that of EG.5.1. Additionally, four clinically available antivirals were effective against BA.2.86. Although the fusogenicity of BA.2.86 spike is similar to that of the parental BA.2 spike, the intrinsic pathogenicity of BA.2.86 in hamsters was significantly lower than that of BA.2. Since the growth kinetics of BA.2.86 are significantly lower than those of BA.2 both in vitro and in vivo, the attenuated pathogenicity of BA.2.86 is likely due to its decreased replication capacity. These findings uncover the features of BA.2.86, providing insights for control and treatment.

• MeSH
• Amino Acids MeSH
• COVID-19 * MeSH
• Kinetics MeSH
• Cricetinae MeSH
• Mutation MeSH
• SARS-CoV-2 genetics   MeSH
• Animals MeSH
• Check Tag
• Cricetinae MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH

Background:Brucella microti is a pathogen of rodents and wild mammals. Here, we report the first probable infection with B. microti in a mammalogist. Materials and Methods: In the study, we provided complete clinical description as well as laboratory analysis of probable human infection caused by B. microti. Results: Considering the clinical course of the infection, the obvious epidemiological link (a bite by an infected rodent), the isolation of a pathogen from a sick vole that was affected by clinical infection with B. microti, and the specific serological response (slow agglutination test) in human patient, we can conclude that the human disease described here was probably caused by B. microti, an emerging bacterial pathogen transmitted by rodents. Conclusion: Rodents and other wildlife need to be monitored not only for established zoonotic agents such as hantaviruses, lymphocytic choriomeningitis virus, Leptospira spp., Francisella tularensis, but also for Brucella microti and other atypical rodent-borne brucellae.

• MeSH
• Arvicolinae microbiology   MeSH
• Bacteria MeSH
• Brucella * MeSH
• Animals, Wild MeSH
• Rodentia MeSH
• Humans MeSH
• Animals MeSH
• Check Tag
• Humans MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH

In late 2022, SARS-CoV-2 Omicron subvariants have become highly diversified, and XBB is spreading rapidly around the world. Our phylogenetic analyses suggested that XBB emerged through the recombination of two cocirculating BA.2 lineages, BJ.1 and BM.1.1.1 (a progeny of BA.2.75), during the summer of 2022. XBB.1 is the variant most profoundly resistant to BA.2/5 breakthrough infection sera to date and is more fusogenic than BA.2.75. The recombination breakpoint is located in the receptor-binding domain of spike, and each region of the recombinant spike confers immune evasion and increases fusogenicity. We further provide the structural basis for the interaction between XBB.1 spike and human ACE2. Finally, the intrinsic pathogenicity of XBB.1 in male hamsters is comparable to or even lower than that of BA.2.75. Our multiscale investigation provides evidence suggesting that XBB is the first observed SARS-CoV-2 variant to increase its fitness through recombination rather than substitutions.

• MeSH
• COVID-19 * MeSH
• Phylogeny MeSH
• Spike Glycoprotein, Coronavirus genetics   MeSH
• Cricetinae MeSH
• Humans MeSH
• Recombination, Genetic MeSH
• SARS-CoV-2 genetics   MeSH
• Animals MeSH
• Check Tag
• Cricetinae MeSH
• Humans MeSH
• Male MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH

In late 2022, various Omicron subvariants emerged and cocirculated worldwide. These variants convergently acquired amino acid substitutions at critical residues in the spike protein, including residues R346, K444, L452, N460, and F486. Here, we characterize the convergent evolution of Omicron subvariants and the properties of one recent lineage of concern, BQ.1.1. Our phylogenetic analysis suggests that these five substitutions are recurrently acquired, particularly in younger Omicron lineages. Epidemic dynamics modelling suggests that the five substitutions increase viral fitness, and a large proportion of the fitness variation within Omicron lineages can be explained by these substitutions. Compared to BA.5, BQ.1.1 evades breakthrough BA.2 and BA.5 infection sera more efficiently, as demonstrated by neutralization assays. The pathogenicity of BQ.1.1 in hamsters is lower than that of BA.5. Our multiscale investigations illuminate the evolutionary rules governing the convergent evolution for known Omicron lineages as of 2022.

• MeSH
• Biological Assay MeSH
• COVID-19 * MeSH
• Phylogeny MeSH
• Cricetinae MeSH
• Antibodies, Neutralizing MeSH
• Antibodies, Viral MeSH
• SARS-CoV-2 genetics   MeSH
• Amino Acid Substitution MeSH
• Animals MeSH
• Check Tag
• Cricetinae MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH

As presently defined, the tapeworm genus Andrya Railliet, 1895 (Cyclophyllidea: Anoplocephalidae sensu stricto) includes the type species A. rhopalocephala (Riehm, 1881) in hares of the genus Lepus Linnaeus (Leporidae) in western Eurasia and four species in cricetid (Neotominae, Sigmodontinae) and octodontid rodents in North and South America. The host range of Andrya is puzzling, because it is the only genus of anoplocephalid (s. s.) cestodes parasitising both rodents and lagomorphs. The present morphological analysis shows that the American species of Andrya share multiple consistent features, in which they differ from those of A. rhopalocephala and the morphologically related Neandrya cuniculi (Blanchard, 1891). The main differences concern the position of the uterus with respect to the longitudinal osmoregulatory canals and testes. Consequently, a new genus Andryoides gen. n. is proposed for the American species, resulting in the following combinations: Andryoides neotomae (Voge, 1946) comb. n. (type species), Andryoides octodonensis (Babero et Cattan, 1975) comb. n., Andryoides vesicula (Haverkost et Gardner, 2010) comb. n. and Andryoides boliviensis (Haverkost et Gardner, 2010) comb. n. However, A. boliviensis is regarded here as a junior synonym of A. vesicula (new synonymy). The present study also defines the morphological key features for all the valid genera of cestodes of the family Anoplocephalidae (s. s.), and discusses the phylogenetic affinities and historical biogeography of Andryoides and other endemic American anoplocephalid cestodes.

• MeSH
• Arvicolinae MeSH
• Cestoda * MeSH
• Cestode Infections * epidemiology   veterinary   MeSH
• Phylogeny MeSH
• Rodentia MeSH
• Mammals MeSH
• Sigmodontinae MeSH
• Animals MeSH
• Check Tag
• Female MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH