Stenotrophomonas maltophilia, Achromobacter xylosoxidans, and Burkholderia cenocepacia are considered emerging pathogens classified as a public health problem due to extensive antimicrobial resistance. Therefore, the discovery of new therapeutic strategies has become crucial. This study aimed to evaluate the antimicrobial activity of gallic acid and methyl gallate against non-fermenting bacteria. The study included five clinical isolates of Stenotrophomonas maltophilia, Achromobacter xylosoxidans, and Burkholderia cenocepacia. The minimum inhibitory concentrations of gallic acid and methyl gallate were determined by the broth microdilution method. Growth curves, metabolic activity, and biofilm formation of each bacterial strain in the presence or absence of phenolic compounds were performed. Finally, the therapeutic efficacy of the compounds was evaluated using an in vivo model. Gallic acid and methyl gallate showed antibacterial activity against bacterial strains in a concentration range of 64 to 256 μg/mL, both compounds reduced bacterial growth and metabolic activity of the strains, even at subinhibitory concentrations. Only, methyl gallate exhibited activity to inhibit the formation of bacterial biofilms. Moreover, gallic acid and methyl gallate increased larval survival by up to 60% compared to 30% survival of untreated larvae in a bacterial infection model in Galleria mellonella. Our results highlight the potential of gallic acid and methyl gallate as therapeutic alternatives for infections by emerging non-fermentative bacteria.
UNLABELLED: The Aspergillus genus encompasses a diverse array of species, some of which are opportunistic pathogens. Traditionally, human aspergillosis has primarily been linked to a few Aspergillus species, predominantly A. fumigatus. Changes in epidemiology and advancements in molecular techniques have brought attention to less common and previously unrecognized pathogenic cryptic species. Despite the taxonomic recognition of many cryptic species in section Terrei, their virulence potential and clinical implications, compared to A. terreus sensu stricto, remain poorly understood. Hence, the current study utilized the alternative in vivo model Galleria mellonella to evaluate the virulence potential of 19 accepted Aspergillus species in section Terrei, classified into three series (major phylogenetic clades): Terrei, Nivei, and Ambigui. Analyzing the median survival rates of infected larvae of all species in each series revealed that series Ambigui has a significantly lower virulence compared to series Terrei and Nivei. Taking a closer look at series Terrei and Nivei revealed a trend of survival within each clade, dividing the species into two groups: highly virulent (up to 72 h survival) and less virulent (up to 144 h survival). Histological observation, considering fungal distribution and filamentation, further supported this assessment, revealing increased distribution and hyphal formation in virulent species. Additionally, the susceptibility profile of conventional antifungals was determined, revealing an increased azole minimum inhibitory concentration for some tested cryptic species such as A. niveus and A. iranicus. Our results highlight the importance of cryptic species identification, as they can exhibit different levels of virulence and show reduced antifungal susceptibility. IMPORTANCE: With changing fungal epidemiology and an increasingly vulnerable population, cryptic Aspergillus species are emerging as human pathogens. Their diversity and clinical relevance remain underexplored, with some species showing reduced antifungal susceptibility and higher virulence, highlighting the need for better preparedness in clinical practice. Using the Galleria mellonella model, we assessed the virulence of Aspergillus species of section Terrei, including cryptic and non-cryptic species, across three series Terrei, Nivei, and Ambigui. The results revealed significant virulence variation among the series, with some cryptic species displaying high virulence. Histological analysis confirmed increased hyphal formation and fungal spread in the more virulent species. Additionally, elevated azole minimum inhibitory concentrations were also observed in certain cryptic species. This study presents novel insights into the pathogenicity of Aspergillus section Terrei, emphasizing the critical importance of accurately identifying cryptic species due to their diverse virulence potential and antifungal resistance, which may have substantial clinical implications.
- MeSH
- antifungální látky farmakologie MeSH
- Aspergillus * patogenita klasifikace účinky léků genetika MeSH
- aspergilóza * mikrobiologie MeSH
- fylogeneze MeSH
- larva mikrobiologie MeSH
- lidé MeSH
- modely nemocí na zvířatech MeSH
- můry * mikrobiologie MeSH
- virulence MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
Toxocara canis is a widespread parasite of canids with a wide range of paratenic hosts, but also one of the overlooked agents causing nervous system infections of humans. Previous experimental infections of mice demonstrated the impact of high infection doses of larvae on neurobehavioral disorders and pathological changes. In contrast to previous studies, we aimed to investigate the long-term (up to 100 weeks) impact of low- to high-dose infection in mice. We focused on their physical condition, motor skills, and the accompanying pathologies in the brain. Three groups of BALB/c mice were infected with 10, 100, and 1000 T. canis larvae/mouse and specific anti-T. canis excretory-secretory antigens immunoglobulin G antibody response, general condition, and motor skills were tested in defined intervals within 100 weeks after infection. The number of larvae in selected organs was assessed and the pathological changes in the brain were studied histologically. As a result, subtle to severe impairments in general condition and motor skills were detected, with generally earlier onsets occurring the higher the infection dose was. The specific immunoglobulin G antibody levels corresponding to the infection dose were detected in all infected groups. Necrosis, cellular infiltrations, and foamy cells developed in moderate- and high-infection dose mice, in contrast with hemorrhages detected in all groups. This study demonstrated the long-term negative impact of T. canis infection on the paratenic host, particularly at moderate and high infectious doses. Although pathological changes in the brain were observed even in low-infection dose mice, their physical and motor condition was comparable to the control group.
- MeSH
- imunoglobulin G * krev MeSH
- larva MeSH
- modely nemocí na zvířatech MeSH
- mozek * parazitologie patologie MeSH
- myši inbrední BALB C * MeSH
- myši MeSH
- protilátky helmintové * krev MeSH
- Toxocara canis * imunologie MeSH
- toxokaróza * parazitologie MeSH
- zvířata MeSH
- Check Tag
- mužské pohlaví MeSH
- myši MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
Amphotericin B (AmB) is one of the most effective antifungal drugs, with a strong, dose-dependent activity against most Candida and Aspergillus species responsible for life-threatening infections. However, AmB is severely toxic, which hinders its broad use. In this proof-of-concept study, we demonstrate that prodrugging AmB considerably decreases AmB toxicity without affecting its fungicidal activity. For this purpose, we modified the AmB structure by attaching a designer phosphate promoiety, thereby switching off its mode of action and preventing its toxic effects. The original fungicidal activity of AmB was then restored upon prodrug activation by host plasma enzymes. These AmB prodrugs showed a safer toxicity profile than commercial AmB deoxycholate in Candida and Aspergillus species and significantly prolonged larval survival of infected Galleria mellonella larvae. Based on these findings, prodrugging toxic antifungals may be a viable strategy for broadening the antifungal arsenal, opening up opportunities for targeted prodrug design.
- MeSH
- amfotericin B * farmakologie MeSH
- antifungální látky * farmakologie chemie chemická syntéza MeSH
- Aspergillus účinky léků MeSH
- Candida účinky léků MeSH
- larva účinky léků MeSH
- mikrobiální testy citlivosti * MeSH
- molekulární struktura MeSH
- můry účinky léků MeSH
- prekurzory léčiv * farmakologie chemie chemická syntéza MeSH
- vztah mezi dávkou a účinkem léčiva MeSH
- vztahy mezi strukturou a aktivitou MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
Neurofibromatosis type 2 (NF-2) is a dominantly inherited genetic disorder that results from variants in the tumor suppressor gene, neurofibromin 2 (NF2). Here, we report the generation of a conditional zebrafish model of neurofibromatosis established by inducible genetic knockout of nf2a/b, the zebrafish homologs of human NF2. Analysis of nf2a and nf2b expression revealed ubiquitous expression of nf2b in the early embryo, with overlapping expression in the neural crest and its derivatives and in the cranial mesenchyme. In contrast, nf2a displayed lower expression levels. Induction of nf2a/b knockout at early stages increased the proliferation of larval Schwann cells and meningeal fibroblasts. Subsequently, in adult zebrafish, nf2a/b knockout triggered the development of a spectrum of tumors, including vestibular Schwannomas, spinal Schwannomas, meningiomas and retinal hamartomas, mirroring the tumor manifestations observed in patients with NF-2. Collectively, these findings highlight the generation of a novel zebrafish model that mimics the complexities of the human NF-2 disorder. Consequently, this model holds significant potential for facilitating therapeutic screening and elucidating key driver genes implicated in NF-2 onset.
- MeSH
- dánio pruhované * genetika embryologie MeSH
- geneticky modifikovaná zvířata MeSH
- genový knockout * MeSH
- larva metabolismus MeSH
- lidé MeSH
- modely nemocí na zvířatech * MeSH
- neurofibromatóza 2 genetika patologie metabolismus MeSH
- neurofibromatózy genetika patologie metabolismus MeSH
- neurofibromin 2 * genetika metabolismus nedostatek MeSH
- proliferace buněk MeSH
- proteiny dánia pruhovaného * genetika metabolismus nedostatek MeSH
- Schwannovy buňky metabolismus patologie MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
Bacillus thuringiensis (Bt) is known for its Cry and Vip3A pesticidal proteins with high selectivity to target pests. Here, we assessed the potential of a novel neotropical Bt strain (UFT038) against six lepidopteran pests, including two Cry-resistant populations of fall armyworm, Spodoptera frugiperda. We also sequenced and analyzed the genome of Bt UFT038 to identify genes involved in insecticidal activities or encoding other virulence factors. In toxicological bioassays, Bt UFT038 killed and inhibited the neonate growth in a concentration-dependent manner. Bt UFT038 and HD-1 were equally toxic against S. cosmioides, S. frugiperda (S_Bt and R_Cry1 + 2Ab populations), Helicoverpa zea, and H. armigera. However, larval growth inhibition results indicated that Bt UFT038 was more toxic than HD-1 to S. cosmioides, while HD-1 was more active against Chrysodeixis includens. The draft genome of Bt UFT038 showed the cry1Aa8, cry1Ac11, cry1Ia44, cry2Aa9, cry2Ab35, and vip3Af5 genes. Besides this, genes encoding the virulence factors (inhA, plcA, piplC, sph, and chi1-2) and toxins (alo, cytK, hlyIII, hblA-D, and nheA-C) were also identified. Collectively, our findings reveal the potential of the Bt UFT038 strain as a source of insecticidal genes against lepidopteran pests, including S. cosmioides and S. frugiperda.
- MeSH
- Bacillus thuringiensis * genetika metabolismus MeSH
- bakteriální proteiny genetika metabolismus MeSH
- biologická kontrola škůdců MeSH
- endotoxiny metabolismus MeSH
- faktory virulence metabolismus MeSH
- Glycine max MeSH
- hemolyziny genetika metabolismus farmakologie MeSH
- insekticidy * farmakologie metabolismus MeSH
- larva MeSH
- lidé MeSH
- můry * MeSH
- novorozenec MeSH
- Spodoptera metabolismus MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- novorozenec MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
BACKGROUND: Regenerative medicine and transplantation science continuously seek methods to circumvent immune-mediated rejection and promote tissue regeneration. Sertoli cells, with their inherent immunoprotective properties, emerge as pivotal players in this quest. However, whether Sertoli cells can play immunomodulatory role in tadpole tail regeneration and can thus benefit the regeneration process are needed to be discovered. METHODS: Immature Sertoli cells from Xenopus tropicalis (XtiSCs) were transplanted into X. tropicalis tadpoles, followed by the amputation of the final third of their tails. We assessed the migration of XtiSCs, tail regeneration length, muscle degradation and growth, and macrophage counts across various regions including the entire tail, tail trunk, injection site, and regeneration site. The interactions between XtiSCs and macrophages were examined using a confocal microscope. To deplete macrophages, clodronate liposomes were administered prior to the transplantation of XtiSCs, while the administration of control liposomes acted as a negative control. Student's t-test was used to compare the effects of XtiSCs injection to those of a 2/3PBS injection across groups with no liposomes, control liposomes, and clodronate liposomes. RESULTS: XtiSCs have excellent viability after transplantation to tadpole tail and remarkable homing capabilities to the regeneration site after tail amputation. XtiSCs injection increased macrophage numbers at 3 days post-amputation and 5 days post-amputation in the tail trunk, specifically at the injection site and at the regenerated tail, in a macrophage depleted environment (clodronate-liposome injection). What's more, XtiSCs injection decreased muscle fibers degradation significantly at 1 day post-amputation and facilitated new muscle growth significantly at 3 days post-amputation. In addition, whole-mount immunostaining showed that some XtiSCs co-localized with macrophages. And we observed potential mitochondria transport from XtiSCs to macrophages using MitoTracker staining in tadpole tail. CONCLUSIONS: Our study delineates the novel role of XtiSCs in facilitating muscle regeneration post tadpole tail amputation, underscoring a unique interaction with macrophages that is crucial for regenerative success. This study not only highlights the therapeutic potential of Sertoli cells in regenerative medicine but also opens avenues for clinical translation, offering insights into immunoregulatory strategies that could enhance tissue regeneration and transplant acceptance.
- MeSH
- imunomodulace MeSH
- larva * MeSH
- makrofágy * metabolismus imunologie MeSH
- ocas MeSH
- regenerace * MeSH
- Sertoliho buňky * cytologie metabolismus MeSH
- Xenopus * MeSH
- zvířata MeSH
- Check Tag
- mužské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
We investigated gene expression patterns in Lutzomyia and Phlebotomus sand fly vectors of leishmaniases. Using quantitative PCR, we assessed the expression stability of potential endogenous control genes commonly used in dipterans. We analyzed Lutzomyia longipalpis and Phlebotomus papatasi samples from L3 and L4 larval stages, adult sand flies of different sexes, diets, dsRNA injection, and Leishmania infection. Six genes were evaluated: actin, α-tubulin, GAPDH, 60 S ribosomal proteins L8 and L32 (RiboL8 and RiboL32), and elongation factor 1-α (EF1-α). EF1-α was among the most stably expressed along with RiboL8 in L. longipalpis larvae and RiboL32 in adults. In P. papatasi, EF1-α and RiboL32 were the top in larvae, while EF1-α and actin were the most stable in adults. RiboL8 and actin were the most stable genes in dissected tissues and infected guts. Additionally, five primer pairs designed for L. longipalpis or P. papatasi were effective in PCR with Lutzomyia migonei, Phlebotomus duboscqi, Phlebotomus perniciosus, and Sergentomyia schwetzi cDNA. Furthermore, L. longipalpis RiboL32 and P. papatasi α-tubulin primers were suitable for qPCR with cDNA from the other four species. Our research provides tools to enhance relative gene expression studies in sand flies, facilitating the selection of endogenous control for qPCR.
- MeSH
- esenciální geny * MeSH
- hmyz - vektory genetika MeSH
- hmyzí geny MeSH
- larva genetika MeSH
- Leishmania genetika MeSH
- Phlebotomus * genetika MeSH
- Psychodidae genetika MeSH
- stanovení celkové genové exprese metody MeSH
- zvířata MeSH
- Check Tag
- mužské pohlaví MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
Po koupání v přírodních nádržích se v létě 2023 na několika místech v České republice objevila u lidí, zejména u dětí, cerkáriová dermatitida. Jde o imunitní reakci na průnik larvy motolice – ptačí krevničky – do kůže nesprávného hostitele. Mezi hostiteli parazita jsou plicnatí vodní plži – plovatky a okružáci – a definitivní hostitelé vodní ptáci, zejména kachny. Hodiny až dny po koupání se vyvine svědivá vyrážka, následně s puchýřky. Hojení trvá dva týdny, symptomy lze tišit antihistaminiky. Čisté vodní nádrže bez plžů jsou bezpečné a v některých přírodních nádržích po nahlášení onemocnění přistoupila samospráva nebo provozovatel koupaliště k fyzické likvidaci plžů.
After swimming in natural reservoirs in the summer of 2023, cercarial dermatitis or Swimmer's itch appeared in several places in the Czech Republic in people, especially in children. It is an immune reaction to the penetration of a fluke larva – a bird blood fluke into the skin of the wrong host. The intermediate hosts of the parasite are pulmonate aquatic gas- tropods – pond snails and ramshorns – and the definitive hosts are waterfowl, especially ducks. An itchy rash develops hours to days after bathing, followed by blisters. Healing takes two weeks symptoms can be calmed by antihistamines. Clean water reservoirs without snails are safe, and in some natural ones, after the disease has been reported, the municipality or the operator of the swimming pool proceeded to physically dispose of the snails.
- MeSH
- cerkárie * patogenita MeSH
- dítě MeSH
- interakce hostitele a parazita MeSH
- lidé MeSH
- parazitární onemocnění kůže * diagnóza prevence a kontrola terapie MeSH
- rybníky MeSH
- Check Tag
- dítě MeSH
- lidé MeSH
- Geografické názvy
- Česká republika MeSH
Introduction. The fungal pathogen Aspergillus fumigatus can induce prolonged colonization of the lungs of susceptible patients, resulting in conditions such as allergic bronchopulmonary aspergillosis and chronic pulmonary aspergillosis.Hypothesis. Analysis of the A. fumigatus secretome released during sub-lethal infection of G. mellonella larvae may give an insight into products released during prolonged human colonisation.Methodology.Galleria mellonella larvae were infected with A. fumigatus, and the metabolism of host carbohydrate and proteins and production of fungal virulence factors were analysed. Label-free qualitative proteomic analysis was performed to identify fungal proteins in larvae at 96 hours post-infection and also to identify changes in the Galleria proteome as a result of infection.Results. Infected larvae demonstrated increasing concentrations of gliotoxin and siderophore and displayed reduced amounts of haemolymph carbohydrate and protein. Fungal proteins (399) were detected by qualitative proteomic analysis in cell-free haemolymph at 96 hours and could be categorized into seven groups, including virulence (n = 25), stress response (n = 34), DNA repair and replication (n = 39), translation (n = 22), metabolism (n = 42), released intracellular (n = 28) and cellular development and cell cycle (n = 53). Analysis of the Gallerial proteome at 96 hours post-infection revealed changes in the abundance of proteins associated with immune function, metabolism, cellular structure, insect development, transcription/translation and detoxification.Conclusion. Characterizing the impact of the fungal secretome on the host may provide an insight into how A. fumigatus damages tissue and suppresses the immune response during long-term pulmonary colonization.
- MeSH
- Aspergillus fumigatus * metabolismus MeSH
- aspergilóza mikrobiologie metabolismus MeSH
- faktory virulence metabolismus MeSH
- fungální proteiny * metabolismus genetika MeSH
- hemolymfa mikrobiologie metabolismus MeSH
- larva * mikrobiologie MeSH
- můry * mikrobiologie MeSH
- proteom analýza MeSH
- proteomika MeSH
- sekretom metabolismus MeSH
- virulence MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
