Cyanobacteria are prokaryotic organisms characterised by their complex structures and a wide range of pigments. With their ability to fix CO2, cyanobacteria are interesting for white biotechnology as cell factories to produce various high-value metabolites such as polyhydroxyalkanoates, pigments, or proteins. White biotechnology is the industrial production and processing of chemicals, materials, and energy using microorganisms. It is known that exposing cyanobacteria to low levels of stressors can induce the production of secondary metabolites. Understanding of this phenomenon, known as hormesis, can involve the strategic application of controlled stressors to enhance the production of specific metabolites. Consequently, precise measurement of cyanobacterial viability becomes crucial for process control. However, there is no established reliable and quick viability assay protocol for cyanobacteria since the task is challenging due to strong interferences of autofluorescence signals of intercellular pigments and fluorescent viability probes when flow cytometry is used. We performed the screening of selected fluorescent viability probes used frequently in bacteria viability assays. The results of our investigation demonstrated the efficacy and reliability of three widely utilised types of viability probes for the assessment of the viability of Synechocystis strains. The developed technique can be possibly utilised for the evaluation of the importance of polyhydroxyalkanoates for cyanobacterial cultures with respect to selected stressor-repeated freezing and thawing. The results indicated that the presence of polyhydroxyalkanoate granules in cyanobacterial cells could hypothetically contribute to the survival of repeated freezing and thawing.
- MeSH
- fluorescence MeSH
- fluorescenční barviva * metabolismus chemie MeSH
- fyziologický stres * MeSH
- mikrobiální viabilita * MeSH
- polyhydroxyalkanoáty metabolismus MeSH
- průtoková cytometrie * MeSH
- sinice metabolismus fyziologie MeSH
- Synechocystis * metabolismus MeSH
- Publikační typ
- časopisecké články MeSH
Alexander disease (AxD) is a rare and severe neurodegenerative disorder caused by mutations in glial fibrillary acidic protein (GFAP). While the exact disease mechanism remains unknown, previous studies suggest that mutant GFAP influences many cellular processes, including cytoskeleton stability, mechanosensing, metabolism, and proteasome function. While most studies have primarily focused on GFAP-expressing astrocytes, GFAP is also expressed by radial glia and neural progenitor cells, prompting questions about the impact of GFAP mutations on central nervous system (CNS) development. In this study, we observed impaired differentiation of astrocytes and neurons in co-cultures of astrocytes and neurons, as well as in neural organoids, both generated from AxD patient-derived induced pluripotent stem (iPS) cells with a GFAPR239C mutation. Leveraging single-cell RNA sequencing (scRNA-seq), we identified distinct cell populations and transcriptomic differences between the mutant GFAP cultures and a corrected isogenic control. These findings were supported by results obtained with immunocytochemistry and proteomics. In co-cultures, the GFAPR239C mutation resulted in an increased abundance of immature cells, while in unguided neural organoids and cortical organoids, we observed altered lineage commitment and reduced abundance of astrocytes. Gene expression analysis revealed increased stress susceptibility, cytoskeletal abnormalities, and altered extracellular matrix and cell-cell communication patterns in the AxD cultures, which also exhibited higher cell death after stress. Overall, our results point to altered cell differentiation in AxD patient-derived iPS-cell models, opening new avenues for AxD research.
- MeSH
- Alexanderova nemoc * genetika patologie metabolismus MeSH
- astrocyty * metabolismus patologie MeSH
- buněčná diferenciace * fyziologie MeSH
- gliový fibrilární kyselý protein * metabolismus genetika MeSH
- indukované pluripotentní kmenové buňky * metabolismus MeSH
- kokultivační techniky MeSH
- kultivované buňky MeSH
- lidé MeSH
- mutace MeSH
- nervové kmenové buňky metabolismus MeSH
- neurony metabolismus patologie MeSH
- organoidy metabolismus patologie MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
Bioethanol production from lignocellulosic materials is hindered by the high costs of pretreatment and the enzymes. The present study aimed to evaluate whether co-cultivation of four selected cellulolytic fungi yields higher cellulase and xylanase activities compared to the monocultures and to investigate whether the enzymes from the co-cultures yield higher saccharification on selected plant materials without thermo-chemical pretreatment. The fungal isolates, Trichoderma reesei F118, Penicillium javanicum FS7, Talaromyces sp. F113, and Talaromyces pinophilus FM9, were grown as monocultures and binary co-cultures under submerged conditions for 7 days. The cellulase and xylanase activities of the culture filtrates were measured, and the culture filtrates were employed for the saccharification of sugarcane leaves, Guinea grass leaves, and water hyacinth stems and leaves. Total reducing sugars and individual sugars released from each plant material were quantified. The co-culture of Talaromyces sp. F113 with Penicillium javanicum FS7 and of T. reesei F118 with T. pinophilus FM9 produced significantly higher cellulase activities compared to the corresponding monocultures whereas no effect was observed on xylanase activities. Overall, the highest amounts of total reducing sugars and individual sugars were obtained from Guinea grass leaves saccharified with the co-culture of T. reesei F118 with T. pinophilus FM9, yielding 63.5% saccharification. Guinea grass leaves were found to be the most susceptible to enzymatic saccharification without pre-treatment, while water hyacinth stems and leaves were the least. Accordingly, the study suggests that fungal co-cultivation could be a promising approach for the saccharification of lignocellulosic materials for bioethanol production.
- MeSH
- celulasa * metabolismus MeSH
- endo-1,4-beta-xylanasy metabolismus MeSH
- ethanol metabolismus MeSH
- Hypocreales enzymologie metabolismus růst a vývoj MeSH
- kokultivační techniky * MeSH
- lignin * metabolismus MeSH
- listy rostlin mikrobiologie MeSH
- Penicillium * enzymologie metabolismus růst a vývoj MeSH
- Saccharum * mikrobiologie metabolismus MeSH
- Talaromyces * enzymologie metabolismus růst a vývoj MeSH
- Publikační typ
- časopisecké články MeSH
Halophilic bacteria are extremophiles that thrive in saline environment. Their ability to withstand such harsh conditions makes them an ideal choice for industrial applications such as lignocellulosic biomass degradation. In this study, a halophilic bacterium with the ability to produce extracellular cellulases and hemicellulases, designated as Nesterenkonia sp. CL21, was isolated from mangrove sediment in Tanjung Piai National Park, Malaysia. Thus far, studies on lignocellulolytic enzymes concerning bacterial species under this genus are limited. To gain a comprehensive understanding of its lignocellulose-degrading potential, the whole genome was sequenced using the Illumina NovaSeq 6000 platform. The genome of strain CL21 was assembled into 25 contigs with 3,744,449 bp and a 69.74% GC content and was predicted to contain 3,348 coding genes. Based on taxonomy analysis, strain CL21 shares 73.8 to 82.0% average nucleotide identity with its neighbouring species, below the 95% threshold, indicating its possible status as a distinct species in Nesterenkonia genus. Through in-depth genomic mining, a total of 81 carbohydrate-active enzymes were encoded. Among these, 24 encoded genes were identified to encompass diverse cellulases (GH3), xylanases (GH10, GH11, GH43, GH51, GH127 and CE4), mannanases (GH38 and GH106) and pectinases (PL1, PL9, and PL11). The production of lignocellulolytic enzymes was tested in the presence of several substrates. This study revealed that strain CL21 can produce a diverse array of enzymes which are active at different time points. By combining experimental data with genomic information, the ability of strain CL21 to produce lignocellulolytic enzymes has been elucidated, with potential applications in biorefinery industry.
- MeSH
- bakteriální proteiny genetika metabolismus MeSH
- celulasy genetika metabolismus MeSH
- fylogeneze * MeSH
- genom bakteriální * MeSH
- genomika * MeSH
- geologické sedimenty mikrobiologie MeSH
- glykosidhydrolasy * genetika metabolismus MeSH
- lignin * metabolismus MeSH
- RNA ribozomální 16S genetika MeSH
- sekvenování celého genomu MeSH
- zastoupení bazí MeSH
- Publikační typ
- časopisecké články MeSH
Fibrilácia predsiení (FP) je najčastejšia arytmia v klinickej praxi prispievajúca k zvýšenej morbidite a mortalite. Humorálne biomarkery, ako natriuretické peptidy, troponín, aldosterón, kortizol, kopeptín a apelín, zohrávajú čoraz dôležitejšiu úlo- hu v diagnostike, predikcii prognózy a manažmente FP. Zvýšené hladiny týchto biomarkerov naznačujú nielen poruchu funkcie myokardu a remodeláciu predsiení, ale aj zápalové a prokoagulačné stavy, ktoré ovplyvňujú vývoj a komplikácie FP. Sledovanie hladín biomarkerov poskytuje hlbší náhľad na patofyziologické mechanizmy FP a môže pomôcť pri identi- fikácii pacientov so zvýšeným rizikom komplikácií, ako sú tromboembolické príhody alebo progresia ochorenia. Integrácia biomarkerov do klinickej praxe môže výrazne zlepšiť stratifikáciu rizika, umožniť personalizovanejší prístup k liečbe FP a prispieť k efektívnejšiemu monitorovaniu priebehu ochorenia. Dôkazy o spojitosti medzi biomarkermi a FP sú povzbud- zujúce, avšak sú potrebné ďalšie štúdie na potvrdenie ich klinického využitia v štandardnej starostlivosti o pacientov s týmto závažným ochorením.
Atrial fibrillation (AF) is the most common arrhythmia in clinical practice, contributing to increased morbidity and mortality. Humoral biomarkers, such as natriuretic peptides, troponin, aldosterone, cortisol, copeptin, and apelin, are gaining importance in the diagnosis, prognosis, and management of AF. Elevated levels of these biomarkers indicate not only myocardial dysfunction and atrial remodeling but also inflammatory and procoagulant states that influence the progression and complications of AF. Monitoring biomarker levels provides deeper insight into the pathophysiological mechanisms of AF and can aid in identifying patients at higher risk of complications, such as thromboembolic events or disease progression. Integrating biomarkers into clinical practice can significantly improve risk stratification, facilitate a more personalized approach to AF treatment, and contribute to more effective disease monitoring. Evidence linking biomarkers with AF is promising; however, further studies are needed to confirm their clinical utility in standard care for patients with this serious condition.
- MeSH
- aldosteron MeSH
- apelin MeSH
- arginin vasopresin MeSH
- biologické markery MeSH
- fibrilace síní * diagnóza MeSH
- hydrokortison MeSH
- lidé MeSH
- natriuretické peptidy MeSH
- troponin krev MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- přehledy MeSH
The use of nanoparticles as a delivery system for a specific antigen could solve many limitations of mucosal vaccine applications, such as low immunogenicity, or antigen protection and stabilization. In this study, we tested the ability of nasally administered chitosan nanoparticles loaded with glycoprotein B of murine cytomegalovirus to induce an immune response in an animal model. The choice of chitosan nanoparticle type was made by in vitro evaluation of sorption efficiency and antigen release. Three types of chitosan nanoparticles were prepared: crosslinked with tripolyphosphate, coated with hyaluronic acid, and in complex with polycaprolactone. The hydrodynamic size of the nanoparticles by dynamic light scattering, zeta potential, Fourier transform infrared spectroscopy, scanning electron microscopy, stability, loading efficiency, and release kinetics with ovalbumin were evaluated. Balb/c mice were immunized intranasally using the three-dose protocol with nanoparticles, gB, and adjuvants Poly(I:C) and CpG ODN. Subsequently, the humoral and cell-mediated antigen-specific immune response was determined. On the basis of the properties of the tested nanoparticles, the cross-linked nanoparticles were considered optimal for further investigation. The results show that nanoparticles with Poly(I:C) and with gB alone raised IgG antibody levels above the negative control. In the case of mucosal IgA, only gB alone weakly induced the production of IgA antibodies compared to saline-immunized mice. The number of activated cells increased slightly in mice immunized with nanoparticles and gB compared to those immunized with gB alone or to negative control. The results demonstrated that chitosan nanoparticles could have potential in the development of mucosal vaccines.
- MeSH
- adjuvancia imunologická MeSH
- aplikace intranazální MeSH
- chitosan * chemie MeSH
- glykoproteiny MeSH
- imunizace MeSH
- imunoglobulin A MeSH
- Muromegalovirus * MeSH
- myši inbrední BALB C MeSH
- myši MeSH
- nanočástice * chemie MeSH
- slizniční imunita MeSH
- vakcíny * MeSH
- zvířata MeSH
- Check Tag
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
Using an active targeting approach of chemotherapeutics-loaded nanocarriers (NCs) with monoclonal antibodies is a potential strategy to improve the specificity of the delivery systems and reduce adverse reactions of chemotherapeutic drugs. Specific targeting of the human epidermal growth factor receptor-2 (HER-2), expressed excessively in HER-2-positive breast cancer cells, can be achieved by conjugating NCs with an anti-HER-2 monoclonal antibody. We constructed trastuzumab-conjugated chitosan iodoacetamide-coated NCs containing doxorubicin (Tras-Dox-CHI-IA-NCs) as a tumor-targeted drug delivery system, during the study. Chitosan-iodoacetamide (CHI-IA) was synthesized and utilized to prepare trastuzumab-conjugated NCs (Tras-NCs). The morphology, physicochemical properties, drug loading, drug release, and biological activities of the NCs were elucidated. The Tras-NCs were spherical, with a particle size of approximately 76 nm, and had a positive zeta potential; after incorporating the drug, the size of the Tras-NC increased. A prolonged, 24-h drug release from the NCs was achieved. The Tras-NCs exhibited high cellular accumulation and significantly higher antitumor activity against HER-2-positive breast cancer cells than the unconjugated NCs and the drug solution. Therefore, Tras-Dox-CHI-IA-NCs could be a promising nanocarrier for HER-2-positive breast cancer.
- MeSH
- chitosan * chemie MeSH
- doxorubicin chemie MeSH
- jodacetamid MeSH
- lékové transportní systémy MeSH
- lidé MeSH
- monoklonální protilátky chemie MeSH
- nádory prsu * farmakoterapie MeSH
- nanočástice * chemie MeSH
- nosiče léků chemie MeSH
- trastuzumab MeSH
- Check Tag
- lidé MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
BACKGROUND: Chronic lymphocytic leukemia (CLL) is a common adult leukemia characterized by the accumulation of neoplastic mature B cells in blood, bone marrow, lymph nodes, and spleen. The disease biology remains unresolved in many aspects, including the processes underlying the disease progression and relapses. However, studying CLL in vitro poses a considerable challenge due to its complexity and dependency on the microenvironment. Several approaches are utilized to overcome this issue, such as co-culture of CLL cells with other cell types, supplementing culture media with growth factors, or setting up a three-dimensional (3D) culture. Previous studies have shown that 3D cultures, compared to conventional ones, can lead to enhanced cell survival and altered gene expression. 3D cultures can also give valuable information while testing treatment response in vitro since they mimic the cell spatial organization more accurately than conventional culture. METHODS: In our study, we investigated the behavior of CLL cells in two types of material: (i) solid porous collagen scaffolds and (ii) gel composed of carboxymethyl cellulose and polyethylene glycol (CMC-PEG). We studied CLL cells' distribution, morphology, and viability in these materials by a transmitted-light and confocal microscopy. We also measured the metabolic activity of cultured cells. Additionally, the expression levels of MYC, VCAM1, MCL1, CXCR4, and CCL4 genes in CLL cells were studied by qPCR to observe whether our novel culture approaches lead to increased adhesion, lower apoptotic rates, or activation of cell signaling in relation to the enhanced contact with co-cultured cells. RESULTS: Both materials were biocompatible, translucent, and permeable, as assessed by metabolic assays, cell staining, and microscopy. While collagen scaffolds featured easy manipulation, washability, transferability, and biodegradability, CMC-PEG was advantageous for its easy preparation process and low variability in the number of accommodated cells. Both materials promoted cell-to-cell and cell-to-matrix interactions due to the scaffold structure and generation of cell aggregates. The metabolic activity of CLL cells cultured in CMC-PEG gel was similar to or higher than in conventional culture. Compared to the conventional culture, there was (i) a lower expression of VCAM1 in both materials, (ii) a higher expression of CCL4 in collagen scaffolds, and (iii) a lower expression of CXCR4 and MCL1 (transcript variant 2) in collagen scaffolds, while it was higher in a CMC-PEG gel. Hence, culture in the material can suppress the expression of a pro-apoptotic gene (MCL1 in collagen scaffolds) or replicate certain gene expression patterns attributed to CLL cells in lymphoid organs (low CXCR4, high CCL4 in collagen scaffolds) or blood (high CXCR4 in CMC-PEG).
- MeSH
- buněčné kultury metody MeSH
- chronická lymfatická leukemie * patologie metabolismus MeSH
- gely chemie MeSH
- kolagen * chemie farmakologie MeSH
- lidé MeSH
- polyethylenglykoly * chemie MeSH
- receptory CXCR4 metabolismus MeSH
- sodná sůl karboxymethylcelulosy * chemie farmakologie MeSH
- techniky 3D buněčné kultury metody MeSH
- tkáňové podpůrné struktury * chemie MeSH
- viabilita buněk účinky léků MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
Successful engraftment of skin grafts highly depends on the quality of the wound bed. Good quality of blood vessels near the surface is critical to support the viability of the graft. Ischemic, irradiated scar tissue, bone and tendons will not have the sufficient blood supply. In such situations flaps are to be resorted. However, the flaps also need to have good vascularity over the limbs. The introduction of dermal substitutes has provided a novel method for repairing various severe skin defects. These substitutes act as dermal regenerative templates, which facilitate dermal reconstruction and regeneration. This study was done to ascertain the effectiveness of these substitutes in the treatment of complex wounds. Between January 2022 and June 2023, 20 patients who had complex wounds, which could not be treated with simple skin grafting and who were treated with collagen and elastin matrix and split skin grafting (SSG) were retrospectively studied. The percentage of SSG take as per the records was noted at a 10-day post-operative period. Patient characteristics, comorbidities, duration and outcomes of the treatment were noted. Twenty patients were included in the study. The minimum size of the ulcer was 5 × 4 cm (area of 20 cm2) and the maximum size of the ulcer was 15 × 15 cm (225 cm2). Average take of skin graft was 93.7% at 10th post-operative day. Recurrence at 6 months was nil. The scar quality was assessed by patient and observer at 3 months and 6 months post-operatively. The lower-limb ulcers with compromised surrounding tissue are complex. The major goal in these cases is to do simple surgery and prevent recurrence. The collagen and elastin matrices provide structural support for cellular infiltration, which helps maximize a SSG take and a stable long-term scar.
- MeSH
- bércové vředy patologie terapie MeSH
- elastin terapeutické užití MeSH
- kolagen terapeutické užití MeSH
- lidé středního věku MeSH
- lidé MeSH
- poranění dolní končetiny * patologie terapie MeSH
- průzkumy zdravotní péče metody statistika a číselné údaje MeSH
- transplantace kůže * metody MeSH
- vředy dolních končetin patologie terapie MeSH
- Check Tag
- lidé středního věku MeSH
- lidé MeSH
- mužské pohlaví MeSH
- ženské pohlaví MeSH
- Publikační typ
- klinická studie MeSH
