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Biopriming seeds with beneficial bacteria has potential to enhance seed germination. Therefore, in this investigation, five sulphur-oxidizing bacterial cultures, viz., Pantoea dispersa SOB2, Bacillus velezensis SN06, Bacillus cereus SN20, Bacillus tropicus SN16, and Bacillus megaterium SN11, were evaluated for different plant growth-promoting traits and their impact on Vigna radiata L. (mung bean) and Brassica juncea L. (mustard) seed germination. Among these, three bacterial cultures Pantoea dispersa SOB2, Bacillus velezensis SN06, and Bacillus megaterium SN11 evinced potential for mineral solubilization on solid medium where Pantoea dispersa SOB2 had the maximum solubilization indices-3.06, 5.14, and 2.48 for phosphate, zinc, and potassium respectively. The culture also displayed higher indole acetic acid (113.12 μg/mL), gibberellic acid (162.66 μg/mL), ammonia (5.23 μg/mL), and siderophore (69.53%) production than other bacterial cultures whereas Bacillus cereus SN20 showed maximum exopolysaccharide production (9.26 g/L). Bacterial culture Pantoea dispersa SOB2 significantly ameliorated the germination rate (3.73 no./day) and relative seed germination (208%) of Brassica juncea L., while Bacillus velezensis SN06 and Bacillus cereus SN20 followed with germination rate and relative seed germination of 2.86 no./day and 207%, respectively. Pantoea dispersa SOB2 displayed lowest mean germination time 2.91 days followed by Bacillus megaterium SN11 with 3.19 days. Biopriming with sulphur-oxidizing bacterial cultures, germination parameters of Vigna radiata L. were also markedly improved. Pantoea dispersa SOB2 demonstrated the highest germination rate (6.72 no./day), relative seed germination (115.56%), and minimum mean generation time (1.73 days). Bacillus velezensis SN06 inoculation had a beneficial effect on the seedling growth of Vigna radiata L., whereas Pantoea dispersa SOB2 greatly aided the seedling growth of Brassica juncea L. Results corroborated a prominent positive correlation between seed germination and plant growth-promoting traits. This is the first study on Pantoea dispersa as sulphur oxidizer, displaying plant growth promoting traits and seed germination potential. The potent sulphur-oxidizing bacterial cultures possessing plant growth promoting activities enhanced seed germination under in vitro conditions that could be further explored in field as biofertilizers to enhance the growth and yield of Brassica juncea L. and Vigna radiata L. crop.
- MeSH
- Bacillus * metabolismus MeSH
- Bacteria * metabolismus MeSH
- hořčice rodu Brassica * růst a vývoj mikrobiologie MeSH
- klíčení * MeSH
- kyseliny indoloctové metabolismus MeSH
- oxidace-redukce MeSH
- Pantoea metabolismus MeSH
- regulátory růstu rostlin metabolismus MeSH
- semena rostlinná * mikrobiologie růst a vývoj MeSH
- síra * metabolismus MeSH
- vigna * růst a vývoj mikrobiologie MeSH
- Publikační typ
- časopisecké články MeSH
The intertumoral and intratumoral heterogeneity of colorectal adenocarcinoma (CRC) at the morphologic level is poorly understood. Previously, we identified morphological patterns associated with CRC molecular subtypes and their distinct molecular motifs. Here we aimed to evaluate the heterogeneity of these patterns across CRC. Three pathologists evaluated dominant, secondary, and tertiary morphology on four sections from four different FFPE blocks per tumor in a pilot set of 22 CRCs. An AI-based image analysis tool was trained on these tumors to evaluate the morphologic heterogeneity on an extended set of 161 stage I-IV primary CRCs (n = 644 H&E sections). We found that most tumors had two or three different dominant morphotypes and the complex tubular (CT) morphotype was the most common. The CT morphotype showed no combinatorial preferences. Desmoplastic (DE) morphotype was rarely dominant and rarely combined with other dominant morphotypes. Mucinous (MU) morphotype was mostly combined with solid/trabecular (TB) and papillary (PP) morphotypes. Most tumors showed medium or high heterogeneity, but no associations were found between heterogeneity and clinical parameters. A higher proportion of DE morphotype was associated with higher T-stage, N-stage, distant metastases, AJCC stage, and shorter overall survival (OS) and relapse-free survival (RFS). A higher proportion of MU morphotype was associated with higher grade, right side, and microsatellite instability (MSI). PP morphotype was associated with earlier T- and N-stage, absence of metastases, and improved OS and RFS. CT was linked to left side, lower grade, and better survival in stage I-III patients. MSI tumors showed higher proportions of MU and TB, and lower CT and PP morphotypes. These findings suggest that morphological shifts accompany tumor progression and highlight the need for extensive sampling and AI-based analysis. In conclusion, we observed unexpectedly high intratumoral morphological heterogeneity of CRC and found that it is not heterogeneity per se, but the proportions of morphologies that are associated with clinical outcomes.
- MeSH
- adenokarcinom * patologie genetika mortalita MeSH
- dospělí MeSH
- kolorektální nádory * patologie genetika mortalita MeSH
- lidé středního věku MeSH
- lidé MeSH
- senioři nad 80 let MeSH
- senioři MeSH
- staging nádorů MeSH
- Check Tag
- dospělí MeSH
- lidé středního věku MeSH
- lidé MeSH
- mužské pohlaví MeSH
- senioři nad 80 let MeSH
- senioři MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
Daunomycin is a chemotherapeutic agent widely used for the treatment of leukemia, but its toxicity toward healthy dividing cells limits its clinical use and its production by fermentation. Herein, we describe the development of a specialized cultivation medium for daunomycin production, including a shift to oil rather than sugar as the primary carbon source. This achieved an almost threefold increase in daunomycin yields, reaching 5.5-6.0 g/L. Daunomycin produced in the oil-based medium was predominantly found in the solid sediment, whereas that produced in the sugar-based medium was mostly soluble. The oil-based medium thus induces an autonomous daunomycin-resistance mechanism involving biogenic nanoparticle formation. The characterization of the nanoparticles confirmed the incorporation of iron and daunomycin, indicating that this approach has the potential to mitigate cytotoxicity while improving yields. The presence of proteins associated with iron homeostasis and oxidative stress responses revealed the ability of the production strain to adapt to high iron concentrations. Our findings provide insight into the mechanisms of biogenic nanoparticle formation and the optimization of cultivation processes. Further investigation will help to refine microbial production systems for daunomycin and also broaden the application of similar strategies for the synthesis of other therapeutically important compounds.
- Publikační typ
- časopisecké články MeSH
As organoids and organ-on-chip (OoC) systems move toward preclinical and clinical applications, there is an increased need for method validation. Using a liquid chromatography-mass spectrometry (LC-MS)-based approach, we developed a method for measuring small-molecule drugs and metabolites in the cell medium directly sampled from liver organoids/OoC systems. The LC-MS setup was coupled to an automatic filtration and filter flush system with online solid-phase extraction (SPE), allowing for robust and automated sample cleanup/analysis. For the matrix, rich in, e.g., protein, salts, and amino acids, no preinjection sample preparation steps (protein precipitation, SPE, etc.) were necessary. The approach was demonstrated with tolbutamide and its liver metabolite, 4-hydroxytolbutamide (4HT). The method was validated for analysis of cell media of human stem cell-derived liver organoids cultured in static conditions and on a microfluidic platform according to Food and Drug Administration (FDA) guidelines with regards to selectivity, matrix effects, accuracy, precision, etc. The system allows for hundreds of injections without replacing chromatography hardware. In summary, drug/metabolite analysis of organoids/OoCs can be performed robustly with minimal sample preparation.
- MeSH
- chromatografie kapalinová metody MeSH
- extrakce na pevné fázi MeSH
- hmotnostní spektrometrie metody MeSH
- játra * metabolismus MeSH
- kapalinová chromatografie-hmotnostní spektrometrie MeSH
- knihovny malých molekul analýza metabolismus chemie MeSH
- laboratoř na čipu MeSH
- léčivé přípravky metabolismus analýza MeSH
- lidé MeSH
- organoidy * metabolismus cytologie MeSH
- tolbutamid metabolismus analýza MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- validační studie MeSH
OBJECTIVES: Cisplatin is a widely used anticancer drug for the treatment of many solid cancers. DNA damage is thought to be the key mechanism of cisplatin’s anticancer activity. However, cisplatin may also affect cellular metabolism. The aim of this study was to determine the effect of cisplatin on the types of ATP production (OXPHOS versus glycolysis) and their rate in prostate cancer cells and to determine the potentially protective effect of autophagy and amino acids during cisplatin treatment. We also wanted to investigate the potential synergy between the metabolic effects of cisplatin on ATP production and the inhibition of autophagy. METHODS: Cisplatin treatment can significantly affect the metabolism of cancer cells. Important metabolic pathways can be altered, leading to changes in energy production and nutrient utilization. Autophagy and amino acid pool modulations can serve as protective mechanisms significantly affecting tumor cell survival under metabolic stress caused by anticancer treatment. By enabling the recycling of amino acids, autophagy helps cancer cells maintain cellular homeostasis and overcome nutrient limitations. Thus, inhibition of autophagy could have a supportive effect on the metabolic effects of cisplatin. RESULTS: After cisplatin treatment, ATP production by way of OXPHOS was significantly decreased in 22Rv1 and PC-3 cells. On the other hand, ATP production by glycolysis was not significantly affected in 22Rv1 cells. DU145 cells with dysfunctional autophagy were the most sensitive to cisplatin treatment and showed the lowest ATP production. However, short-term autophagy inhibition (24h) by autophinib or SAR405 in 22Rv1 and PC-3 cells did not alter the effect of cisplatin on ATP production. Levels of some amino acids (arginine, methionine) significantly affected the fitness of cancer cells. CONCLUSION: Persistent defects of autophagy can affect the metabolic sensitivity of cancer cells due to interference with arginine metabolism. Amino acids contained in the culture medium had an impact on the overall effect of cisplatin (Fig. 3, Ref. 38).
Tannase-producing filamentous fungi residing alongside tannin-rich ambient in the Northwest Himalayas were isolated at laboratory conditions and further identified by 18S ribosomal RNA gene sequencing. Five most potent tannase producing strains (EI ≥ 2.0), designated Aspergillus fumigatus AN1, Fusarium redolens AN2, Penicillium crustosum AN3, Penicillium restrictum AN4, and Penicillium commune AN5, were characterized. The strain Penicillium crustosum AN3 exhibited a maximum zone dia (25.66 mm ± 0.38). During solid-state fermentation, a maximal amount of tannase was attained with Penicillium crustosum AN3 using pine needles (substrate) by adopting response surface methodology for culture parameter optimization. Gel filtration chromatography yielded 46.48% of the partially purified enzyme with 3.94-fold of tannase purification. We found two subunits in enzyme-117.76 KDa and 88.51 KDa, respectively, in the SDS-PAGE. Furthermore, the characterization of partially purified tannase revealed a maximum enzyme activity of 8.36 U/mL at 30 °C using a substrate concentration (methyl gallate) of 10 mM. To broaden the knowledge of crude enzyme application, dye degradation studies were subjected to extracellular crude tannase from Penicillium crustosum AN3 where the maximum degradation achieved at a low enzyme concentration (5 ppm).
- MeSH
- barvicí látky metabolismus chemie MeSH
- fermentace MeSH
- fungální proteiny genetika metabolismus izolace a purifikace chemie MeSH
- Fusarium enzymologie genetika MeSH
- fylogeneze MeSH
- houby enzymologie genetika MeSH
- karboxylesterhydrolasy * metabolismus genetika izolace a purifikace chemie MeSH
- kultivační média chemie MeSH
- molekulová hmotnost MeSH
- Penicillium * enzymologie genetika MeSH
- RNA ribozomální 18S genetika MeSH
- stabilita enzymů MeSH
- substrátová specifita MeSH
- teplota MeSH
- Publikační typ
- časopisecké články MeSH
Yeast populations can undergo diversification during their growth and ageing, leading to the formation of different cell-types. Differentiation into two major subpopulations, differing in cell size and density and exhibiting distinct physiological and metabolic properties, was described in planktonic liquid cultures and in populations of colonies growing on semisolid surfaces. Here, we compare stress resistance, metabolism and expression of marker genes in seven differentiated cell subpopulations emerging during cultivation in liquid fermentative or respiratory media and during colony development on the same type of solid media. The results show that the more-dense cell subpopulations are more stress resistant than the less-dense subpopulations under all cultivation conditions tested. On the other hand, respiratory capacity, enzymatic activities and marker gene expression differed more between subpopulations. These characteristics are more influenced by the lifestyle of the population (colony vs. planktonic cultivation) and the medium composition. Only in the population growing in liquid respiratory medium, two subpopulations do not form as in the other conditions tested, but all cells exhibit a range of characteristics of the more-dense subpopulations. This suggests that signals for cell differentiation may be triggered by prior metabolic reprogramming or by an unknown signal from the structured environment in the colony.
Industrial processes generate various quantities of waste that can be contaminated with radionuclides of natural origin (naturally occurring radioactive material ('NORM waste')). The efficient management of this waste is essential for any industries affected by NORM waste generation. To obtain an overview of current practices and approaches in Europe, the IRPA Task Group on NORM conducted a survey among task group members and other experts from European countries. The results revealed significant differences in methods and approaches in the European countries. In many countries, landfills are used to dispose of small- and medium-sized quantities of NORM waste with limited activity concentration. But our survey indicated that despite a uniform reference basis for national legislation in Europe, there are different framework conditions for the disposal of NORM waste in practice. In some countries, the disposal is hampered because the interface between the radiation protection system and the waste regime is not yet clearly regulated. Particular practical problems include the lack of acceptance of waste because of the 'radioactivity' stigma and only vague specifications by the legislators on acceptance obligations of the waste management sector.
- MeSH
- nakládání s odpady * MeSH
- odpadky - odstraňování * MeSH
- průmysl MeSH
- radiační ochrana * MeSH
- Publikační typ
- časopisecké články MeSH
- Geografické názvy
- Evropa MeSH
A novel ultra-high performance chromatography method with multichannel detection that allows fast, sensitive, and robust analysis of an antifungal drug terbinafine and its three main impurities β-terbinafine, (Z)-terbinafine, and 4-methylterbinafine in just 5.0 min has been developed. Analysis of terbinafine is important in pharmaceutical analysis since it enables the detection of its impurities at very low concentrations. In this study, we focused on the development, optimization, and validation of the UHPLC method as well as its subsequent application in the evaluation of terbinafine and its three main impurities in the dissolution medium to reveal the incorporation of terbinafine in two poly(lactic-co-glycolic acid) (PLGA) carriers and testing of the drug release at pH 5.5. PLGA based drug delivery systems such as solid dispersions, thin films, microparticles, and nanoparticles are new favorable ways of terbinafine administration. PLGA features excellent tissue compatibility, biodegradation, and adjustable drug release profile. Our pre-formulation study indicates that poly(acrylic acid) branched PLGA polyester has more suitable properties than tripentaerythritol branched PLGA polyester. Therefore, the former is likely to enable design of a new drug delivery system for topically applied terbinafine that could facilitate its administration and increase patient compliance.
I přes rychlý pokrok v oblasti testování citlivosti k antibiotikům genetickými metodami je fenotypové testování stále důležitou součástí diagnostiky tuberkulózy, především proto, že pro některá novější antibiotika nelze získat spolehlivé výsledky jiným způsobem. Tradičně se stanovení citlivosti kmenů M. tuberculosis provádí proporční metodou na tuhých médiích (např. Löwenstein-Jensenova vaječná půda, nebo agary Middlebrook 7H10 a 7H11). Zde se věnujeme nejdůležitějším faktorům, které ovlivňují spolehlivost a reprodukovatelnost proporční metody, jako jsou například kvalita použitého média, kvalita a správná koncentrace inokula a správná interpretace výsledků. Uvádíme také nejaktuálnější mezinárodně uznávané hodnoty kritické koncentrace pro antituberkulotika doporučovaná Světovou zdravotnickou organizací k léčbě tuberkulózy.
Despite the fast advances in genotypic drug susceptibility testing, phenotypic testing is still an important part of diagnostics of tuberculosis, namely because it is not possible to obtain reliable results for some of the newer antituberculous drugs in any other way. Traditionally, the phenotypic drug susceptibility testing of M. tuberculosis strains is performed by proportion method on solid media (e.g. Löwenstein-Jensen egg based medium or agar based Middlebrook 7H10 and 7H11). Here we focus on the most important factors impacting the reliability and reproducibility of the proportion method,such as media quality, the quality and right concentration of inoculum and correct result interpretation. We also provide the most up-to-date critical concentration values for antituberculous drugs recommended for tuberculosis treatment by the World Health Organisation.
- MeSH
- antibiotická rezistence MeSH
- antituberkulotika aplikace a dávkování klasifikace terapeutické užití MeSH
- kultivační média klasifikace MeSH
- lidé MeSH
- mikrobiální testy citlivosti metody MeSH
- Mycobacterium tuberculosis * izolace a purifikace metabolismus účinky léků MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- přehledy MeSH
