Tailocins are nano-scale phage tail-like protein complexes that can mediate antagonistic interactions between closely related bacterial species. While the capacity to produce R-type tailocin was found widely across Gammaproteobacteria, the production of F-type tailocins seems comparatively rare. In this study, we examined the freshwater isolate, Pragia fontium 24613, which can produce both R- and F-type tailocins. We investigated their inhibition spectrum, focusing on clinically relevant enterobacteria, and identified the associated tailocin gene cluster. Transmission electron microscopy confirmed that inactivation of the tape measure protein within the tailocin cluster disrupted R-tailocin production. Comparative analysis of Budviciaceae gene clusters showed high conservation of R-type tailocin genes, whereas F-type tailocin genes were found in only a few species, with little conservation. Our findings indicate a high prevalence of bacteriocin production among underexplored Enterobacteriales species. Detected tailocins showed potential as antimicrobials targeting clinically significant pathogens.
BACKGROUND: Renal cell carcinoma (RCC) is a disease typified by anomalies in cell metabolism. The function of mitochondria, including subunits of mitochondrial respiratory complex II (CII), in particular SDHB, are often affected. Here we investigated the state and function of CII in RCC patients. METHODS: We evaluated tumour tissue as well as the adjacent healthy kidney tissue of 78 patients with RCC of different histotypes, focusing on their mitochondrial function. As clear cell RCC (ccRCC) is by far the most frequent histotype of RCC, we focused on these patients, which were grouped based on the pathological WHO/ISUP grading system to low- and high-grade patients, indicative of prognosis. We also evaluated mitochondrial function in organoids derived from tumour tissue of 7 patients. RESULTS: ccRCC tumours were characterized by mutated von Hippel-Lindau gene and high expression of carbonic anhydrase IX. We found low levels of mitochondrial DNA, protein and function, together with CII function in ccRCC tumour tissue, but not in other RCC types and non-tumour tissues. Mitochondrial content increased in high-grade tumours, while the function of CII remained low. Tumour organoids from ccRCC patients recapitulated molecular characteristics of RCC tissue. CONCLUSIONS: Our findings suggest that the state of CII, epitomized by its assembly and SDHB levels, deteriorates with the progressive severity of ccRCC. These observations hold the potential for stratification of patients with worse prognosis and may guide the exploration of targeted therapeutic interventions.
- MeSH
- Antigens, Neoplasm MeSH
- Adult MeSH
- Carbonic Anhydrase IX metabolism genetics MeSH
- Carcinoma, Renal Cell * pathology metabolism genetics MeSH
- Middle Aged MeSH
- Humans MeSH
- DNA, Mitochondrial genetics metabolism MeSH
- Mitochondria * metabolism pathology genetics MeSH
- Mutation MeSH
- Von Hippel-Lindau Tumor Suppressor Protein genetics metabolism MeSH
- Kidney Neoplasms * pathology metabolism genetics MeSH
- Electron Transport Complex II * metabolism genetics MeSH
- Aged MeSH
- Succinate Dehydrogenase genetics metabolism MeSH
- Check Tag
- Adult MeSH
- Middle Aged MeSH
- Humans MeSH
- Male MeSH
- Aged MeSH
- Female MeSH
- Publication type
- Journal Article MeSH
Nitro-fatty acids (NO2FAs) are endogenously produced electrophiles and NRF2 activators with therapeutic potential. We developed a synthetic protocol combining a Henry reaction and base-promoted β-elimination, yielding ultrapure regio/stereoisomers of nitro-stearic (NO2SA), nitro-oleic (NO2OA), and conjugated/bis-allylic nitro-linoleic (NO2LA) acids. These were tested for NRF2 pathway activation in bone marrow cells under different oxygen conditions. We observed that 9- and 10-NO2OA, and 10-NO2LA increased NRF2 stabilization under hypoxia, while 9- and 10-NO2OA significantly upregulated Hmox1 and Gclm at all oxygen levels. 9- and 10-NO2OA enhanced HO-1 and GCLM proteins independently of oxygen, while 10-NO2LA was oxygen-dependent, boosting HO-1 under hypoxia and GCLM under ambient conditions. Moreover, 10-NO2OA and 10-NO2LA induced NRF2 nuclear translocation. In contrast, the saturated 10-NO2SA, which has lower electron-acceptor ability, was inactive. In summary, these findings suggest the biological activity of NO2FAs is dependent on oxygen level, which could be used in future research of other oxidative stress-dependent pathways.
- MeSH
- Nitro Compounds * pharmacology chemical synthesis chemistry MeSH
- NF-E2-Related Factor 2 * metabolism MeSH
- Heme Oxygenase-1 metabolism MeSH
- Cell Hypoxia MeSH
- Linoleic Acids chemical synthesis chemistry pharmacology MeSH
- Oxygen metabolism MeSH
- Fatty Acids * pharmacology chemical synthesis chemistry MeSH
- Mice MeSH
- Signal Transduction drug effects MeSH
- Stereoisomerism MeSH
- Animals MeSH
- Check Tag
- Mice MeSH
- Animals MeSH
- Publication type
- Journal Article MeSH
Nedd4-2 E3 ligase regulates Na+ homeostasis by ubiquitinating various channels and membrane transporters, including the epithelial sodium channel ENaC. In turn, Nedd4-2 dysregulation leads to various conditions, including electrolytic imbalance, respiratory distress, hypertension, and kidney diseases. However, Nedd4-2 regulation remains mostly unclear. The present study aims at elucidating Nedd4-2 regulation by structurally characterizing Nedd4-2 and its complexes using several biophysical techniques. Our cryo-EM reconstruction shows that the C2 domain blocks the E2-binding surface of the HECT domain. This blockage, ubiquitin-binding exosite masking by the WW1 domain, catalytic C922 blockage and HECT domain stabilization provide the structural basis for Nedd4-2 autoinhibition. Furthermore, Ca2+-dependent C2 membrane binding disrupts C2/HECT interactions, but not Ca2+ alone, whereas 14-3-3 protein binds to a flexible region of Nedd4-2 containing the WW2 and WW3 domains, thereby inhibiting its catalytic activity and membrane binding. Overall, our data provide key mechanistic insights into Nedd4-2 regulation toward fostering the development of strategies targeting Nedd4-2 function.
- MeSH
- Cryoelectron Microscopy MeSH
- HEK293 Cells MeSH
- Humans MeSH
- Models, Molecular MeSH
- Protein Domains MeSH
- 14-3-3 Proteins * metabolism chemistry MeSH
- Ubiquitination MeSH
- Nedd4 Ubiquitin Protein Ligases * metabolism chemistry genetics ultrastructure MeSH
- Calcium * metabolism MeSH
- Protein Binding MeSH
- Check Tag
- Humans MeSH
- Publication type
- Journal Article MeSH
Východiska: Cholangiocelulární karcinom je agresivní maligní onemocnění s rostoucí incidencí a nepříznivou prognózou, přičemž medián přežití u pokročilého onemocnění činí přibližně 12 měsíců. Standardem léčby zůstává systémová chemoterapie na bázi platinového derivátu, avšak její účinnost je limitovaná. Genetické změny jako mutace IDH1 představují potenciální cíle pro cílenou terapii, zejména u pacientů s intrahepatálním cholangiocelulárním karcinomem. Ivosidenib, perorální inhibitor IDH1, prokázal ve studii fáze III ClarIDHy zlepšení přežití bez progrese onemocnění u pacientů s cholangiocelulárním karcinomem a mutací IDH1. Pozorování: Prezentujeme případ 62letého pacienta s pokročilým cholangiocelulárním karcinomem a mutací IDH1, u něhož na standardní chemoterapii po přechodné stabilizaci došlo k progresi. Pro přítomnost alterace IDH1 byl pacient léčen ivosidenibem v druhé linii. Na této léčbě došlo ke stabilizaci nemoci dle RECIST kritérií. Subjektivně však došlo k významnému kvalitativnímu zlepšení. Pacient dosáhl více než trojnásobně delšího přežití bez progrese v porovnání s přežitím dosaženým v rámci klinické studie, a to bez nutnosti redukce dávky. Závěr: Tento případ potvrzuje význam cílené terapie u pacientů s IDH1-mutovaným cholangiocelulárním karcinomem, kde je nejen dosažena objektivní stabilizace nemoci, ale i výrazné subjektivní zlepšení kvality života. Tento přístup podtrhuje význam molekulárního vyšetření a podporuje využití personalizované medicíny v léčbě vzácných typů nádorů, jako je cholangiocelulární karcinom, i přes relativně nízkou frekvenci objektivních odpovědí.
Background: Cholangiocarcinoma is an aggressive cancer with an increasing incidence and a poor prognosis, typically resulting in a median survival of about 12 months for advanced cases. The standard treatment has been platinum-based systemic chemotherapy, although its effectiveness is often limited. Genetic alterations, such as mutations in the IDH1 gene, offer potential targets for targeted therapies, particularly in patients with intrahepatic cholangiocarcinoma. Ivosidenib, an oral IDH1 inhibitor, has shown improved progression-free survival in patients with IDH1-mutated cholangiocarcinoma, according to phase III ClarIDHy study. Observation: We present the case of a 62-year-old patient diagnosed with advanced cholangiocarcinoma and an IDH1 mutation. The patient initially responded to standard chemotherapy, which led to a temporary stabilisation of the disease; however, progression was noted after 6 months. Given the presence of the IDH1 alteration, the patient was treated with ivosidenib as a second-line therapy. This treatment resulted in disease stabilisation according to RECIST criteria. Subjectively, the patient experienced a significant improvement in the quality of life. The patient achieved more than three times longer progression-free survival than was achieved in the clinical trial, without the need for dose reduction. Conclusion: This case highlights the importance of targeted therapy for patients with IDH1-mutated cholangiocarcinoma. Not only was objective disease stabilisation achieved, but there was also a significant subjective improvement in the quality of life. This underscores the value of molecular testing and supports the use of personalised medicine when treating rare cancer types like cholangiocarcinoma, even in instances where objective responses are relatively uncommon.
- Keywords
- ivosidenib,
- MeSH
- Cholangiocarcinoma diagnostic imaging diagnosis drug therapy complications MeSH
- Molecular Targeted Therapy methods MeSH
- Isocitrate Dehydrogenase antagonists & inhibitors genetics therapeutic use MeSH
- Middle Aged MeSH
- Humans MeSH
- Femoral Neoplasms diagnostic imaging diagnosis radiotherapy secondary MeSH
- Liver Neoplasms drug therapy secondary MeSH
- Tomography, X-Ray Computed methods MeSH
- Check Tag
- Middle Aged MeSH
- Humans MeSH
- Male MeSH
- Publication type
- Case Reports MeSH
Extracellular vesicles can play an important role in the processes occurring after stem cell transplantation, preventing cell apoptosis, stimulating immunological processes, and promoting the synthesis of extracellular matrix. Human follicular fluid (FF) can be a source of a subpopulation of cells with mesenchymal stem cells (MSCs) properties. Moreover these subpopulations of FF cells can differentiate into osteoblasts. In presented studies flow cytometry of ovarian FF cells confirmed positive expression of MSCs markers such as: CD44, CD90, CD105, CD73 and negative expression of a hematopoietic marker: CD45. The CD90+, CD105+, CD45- cell subpopulation has been obtained during magnetic separation using appropriate antibodies conjugated with microbeads. The extracellular vesicles (EVs) secreted by the cells during osteodifferentiation process differed from those secreted by cells culture in the basal medium. Based on the previous and current electron microscopy research, changes in size, number, and shape would support the notion that released EVs could be crucial to the ovarian FF cell subpopulation differentiation process. Osteogenic differentiation has been confirmed via Alizarin red staining. Therefore, follicular fluid (FF) can be a new source of a cell subpopulation with MSC properties, with the cells capable of differentiating into the osteogenic lineage. EVs could play a key role as mediators in tissue regeneration, especially bone tissue regeneration.
- MeSH
- Cell Differentiation * MeSH
- Extracellular Vesicles * ultrastructure metabolism MeSH
- Follicular Fluid * cytology metabolism MeSH
- Cells, Cultured MeSH
- Humans MeSH
- Mesenchymal Stem Cells * cytology metabolism MeSH
- Osteoblasts cytology metabolism MeSH
- Osteogenesis * MeSH
- Flow Cytometry MeSH
- Check Tag
- Humans MeSH
- Female MeSH
- Publication type
- Journal Article MeSH
Neurons rely on the microtubule cytoskeleton to create and maintain their sophisticated cellular architectures. Advances in cryogenic electron microscopy, expansion microscopy, live imaging, and gene editing have enabled novel insights into mechanisms of centrosomal and acentrosomal microtubule nucleation, the key process generating new microtubules. This has paved the way for the functional dissection of distinct microtubule networks that regulate various processes during neuronal development, including neuronal delamination, polarization, migration, maturation, and synapse function. We review recent progress in understanding the molecular concepts of microtubule nucleation, how these concepts underlie neurodevelopmental processes, and pinpoint the open questions. Since microtubules play a pivotal role in axon regeneration within the adult central nervous system, understanding the processes of microtubule nucleation could inform strategies to enhance the regenerative capabilities of neurons in the future.
- MeSH
- Centrosome * metabolism physiology MeSH
- Humans MeSH
- Microtubules * metabolism physiology MeSH
- Neurogenesis * physiology MeSH
- Neurons * physiology metabolism MeSH
- Animals MeSH
- Check Tag
- Humans MeSH
- Animals MeSH
- Publication type
- Journal Article MeSH
- Review MeSH
Recent advances in optical sensing technologies underpin the development of high-performance, surface-sensitive analytical tools capable of reliable and precise detection of molecular targets in complex biological media in non-laboratory settings. Optical fibre sensors guide light to and from a region of interest, enabling sensitive measurements of localized environments. This positions optical fibre sensors as a highly promising technology for a wide range of biochemical and healthcare applications. However, their performance in real-world biological media is often limited by the absence of robust post-modification strategies that provide both high biorecognition and antifouling capabilities. In this study, we present the proof-of-concept antifouling and biorecognition performance of a polymer brush nano-coating synthesized at the sensing region of optical fibre long-period grating (LPG) sensors. Using a newly developed antifouling terpolymer brush (ATB) composed of carboxybetaine methacrylamide, sulfobetaine methacrylamide, and N-(2-hydroxypropyl)methacrylamide, we achieve state-of-the-art antifouling properties. The successful on-fibre ATB synthesis is confirmed through scanning electron microscopy (SEM), fluorescence microscopy, and label-free bio-detection experiments based on antibody-functionalized ATB-coated LPG optical fibres. Despite the challenges in handling optical fibres during polymerization, the resulting nano-coating retains its remarkable antifouling properties upon exposure to blood plasma and enables biorecognition element functionalization. These capabilities are demonstrated through the detection of IgG in buffer and diluted blood plasma using anti-IgG-functionalized ATB-coated sensing regions of LPG fibres in both label-based (fluorescence) and label-free real-time detection experiments. The results show the potential of ATB-coated LPG fibres for use in analytical biosensing applications.
Skin represents the largest organ in the human body, functioning as a protective barrier against environmental factors while playing a critical role in thermoregulation. Acne vulgaris is recognized as the most common dermatological condition affecting adolescents, and if left untreated, it can result in lasting skin damage and associated psychosocial challenges. This study aims to develop innovative polymeric biomaterials that could effectively support the treatment of acne vulgaris. The synthesis of these biomaterials involves the use of polyethylene glycol 6000, sodium alginate, and the antioxidant protein glutathione (GHS) to create polymeric hydrogels. These hydrogels were generated via a UV-mediated crosslinking process. To enhance the functional properties of the hydrogels, zinc oxide microparticles (ZnO), synthesized through a wet precipitation method, were incorporated into the formulations. Characterization of the ZnO was performed using Fourier-Transform Infrared Spectroscopy (FTIR), X-ray Diffraction (XRD), particle sizer analysis, and Scanning Electron Microscopy (SEM). Additionally, the bioactivity of the synthesized materials was evaluated through incubation in media simulating physiological body fluids. The cytotoxic effects of the biomaterials were assessed using an indirect test on mouse fibroblast (L929) cells, in accordance with ISO 10993-5 guidelines. The results of our research indicate that the developed biomaterials exhibit potential as a carrier for active substances, contributing positively to the treatment of acne vulgaris and potentially improving overall skin health.
- MeSH
- Acne Vulgaris drug therapy MeSH
- Alginates chemistry MeSH
- Biocompatible Materials chemistry pharmacology MeSH
- Cell Line MeSH
- Fibroblasts drug effects metabolism MeSH
- Glutathione * metabolism MeSH
- Hydrogels * chemistry MeSH
- Skin * drug effects metabolism MeSH
- Humans MeSH
- Mice MeSH
- Drug Carriers chemistry MeSH
- Zinc Oxide * chemistry pharmacology MeSH
- Regeneration drug effects MeSH
- Spectroscopy, Fourier Transform Infrared MeSH
- Animals MeSH
- Check Tag
- Humans MeSH
- Mice MeSH
- Animals MeSH
- Publication type
- Journal Article MeSH
Apolipoprotein E (APOE) is distributed across various human tissues and plays a crucial role in lipid metabolism. Recent investigations have uncovered an additional facet of APOE's functionality, revealing its role in host defense against bacterial infections. To assess the antibacterial attributes of APOE3 and APOE4, we conducted antibacterial assays using Pseudomonas aeruginosa and Escherichia coli. Exploring the interaction between APOE isoforms and lipopolysaccharides (LPSs) from E. coli, we conducted several experiments, including gel shift assays, CD, and fluorescence spectroscopy. Furthermore, the interaction between APOE isoforms and LPS was further substantiated through atomic resolution molecular dynamics simulations. The presence of LPS induced the aggregation of APOE isoforms, a phenomenon confirmed through specific amyloid staining, as well as fluorescence and electron microscopy. The scavenging effects of APOE3/4 isoforms were studied through both in vitro and in vivo experiments. In summary, our study established that APOE isoforms exhibit binding to LPS, with a more pronounced affinity and complex formation observed for APOE4 compared with APOE3. Furthermore, our data suggest that APOE isoforms neutralize LPS through aggregation, leading to a reduction of local inflammation in experimental animal models. In addition, both isoforms demonstrated inhibitory effects on the growth of P. aeruginosa and E. coli. These findings provide new insights into the multifunctionality of APOE in the human body, particularly its role in innate immunity during bacterial infections.
- MeSH
- Apolipoprotein E3 * metabolism chemistry pharmacology MeSH
- Apolipoprotein E4 * metabolism chemistry pharmacology MeSH
- Escherichia coli metabolism MeSH
- Humans MeSH
- Lipopolysaccharides * metabolism chemistry MeSH
- Mice MeSH
- Protein Isoforms chemistry metabolism MeSH
- Pseudomonas aeruginosa metabolism MeSH
- Molecular Dynamics Simulation MeSH
- Animals MeSH
- Check Tag
- Humans MeSH
- Mice MeSH
- Animals MeSH
- Publication type
- Journal Article MeSH
