indicator compound
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Bacteria have developed different intra- and inter-specific communication mechanisms that involve the production, release, and detection of signaling molecules, because these molecules serve as the autoinducers involved in "quorum sensing" systems. Other communication mechanisms employ volatile signaling molecules that regulate different bacterial processes. The Arthrobacter agilis strain UMCV2 is a plant growth promoting actinobacterium, which induces plant growth and inhibits phytopathogenic fungi by emitting the dimethylhexadecylamine (DMHDA). However, little is known about the effect of this volatile compound on A. agilis UMCV2 itself, as well as on other bacteria. By exposing A. agilis UMCV2 and bacteria of the genus Bacillus and Pseudomonas to different concentrations of DMHDA, this study showed the dose-dependent effects of DMHDA on A. agilis UMCV2 growth, cellular viability, swarming motility, and expression of marker genes of the flagellar apparatus of bacteria. DMHDA was found to also modulate swarming motility of Bacillus sp. ZAP018 and P. fluorescens UM270, but not that of P. aeruginosa PA01. These data indicate that DMHDA is involved in both intra- and inter-specific bacterial interaction.
- MeSH
- Arthrobacter účinky léků růst a vývoj MeSH
- Bacillus účinky léků růst a vývoj MeSH
- methylaminy farmakologie MeSH
- mikrobiální interakce účinky léků MeSH
- pohyb účinky léků MeSH
- Pseudomonas účinky léků růst a vývoj MeSH
- quorum sensing účinky léků MeSH
- těkavé organické sloučeniny farmakologie MeSH
- Publikační typ
- časopisecké články MeSH
Pro stanovení diagnózy tuberkulózy je vedle klinických příznaků, rentgenového vyšetření plic a kožního tuberkulinového testu rozhodujícím vodítkem průkaz původce onemocnění – Mycobacterium tuberculosis přímou mikroskopií z biologického materiálu a kultivací na pevných vaječných půdách a půdách tekutých (Ogawova, Löwensteinova-Jensenova, Šulova půda). Pomalý růst většiny mykobakteriálních druhů je limitujícím faktorem jak při potvrzení etiologie onemocnění, tak při následných laboratorních testech lékové citlivosti a druhové identifikace, na kterých závisí včasné zahájení léčby, stanovení léčebného režimu a protiepidemických opatření. Jednou z metod navržených k urychlení průkazu mykobakterií a vhodnou k zavedení do praxe v diagnostických laboratořích je kultivační metoda BD BBL (Becton Dickinson): BD-1 Becton Drive Franclin Lakes, NJ USA 07417, MGIT (MGIT-Mycobacteria Growth Indikator Tube suplement), který pro detekci iniciální fáze pomnožování mykobakterií v tekuté půdě, tj. v době, kdy ještě nelze růst makrokolonií vizuálně prokázat, využívá fluorescenční technologii. Fluorescenční látka (Tris, -diphenyl-1, 10-phenaltroline ruthenium chloride pentahydrate) je v tomto systému vázána silikonem na dně zkumavek s tekutým kultivačním médiem a úbytek kyslíku provázející růst mykobakterií vyvolává fluorescenci, jejíž intenzita je registrována UV transiluminátorem.
Diagnosis of tuberculosis is based on clinical symptoms, lung X-ray, skin tuberculin test and primarily on the detection of the causative agent Mycobacterium tuberculosis by direct microscopy of biological specimens and culture on solid egg media and in liquid media (Ogawa, Löwenstein-Jensen, Šula). Slow growth of most mycobacterial species is a limiting factor in both the confirmation of etiology and subsequent drug susceptibility tests and species identification that are of crucial relevance to early institution of treatment, selection of treatment regimen and implementation of antiepidemic measures. One of the methods proposed for more rapid detection of mycobacteria and suitable for use in routine diagnostic laboratories is the BD BBL MGIT culture system (Becton Dickinson, 1 Becton Drive, Franklin Lakes, NJ 07417, USA) based on fluorescence detection of the initial phase of mycobacterial multiplication at which macrocolony growth is still not visible. The fluorescent compound Tris, 4,7-diphenyl-1,10-phenalthroline ruthenium chloride pentahydrate is embedded in silicone on the bottom of tubes with liquid culture medium in which growing, actively respiring mycobacteria consume the oxygen and allow the fluorescence to be detected and visualized using a UV transluminator.
- MeSH
- bifenylové sloučeniny analýza MeSH
- látky znečišťující životní prostředí analýza MeSH
- lidé MeSH
- mateřské mléko MeSH
- monitorování životního prostředí MeSH
- odběr biologického vzorku MeSH
- rezidua pesticidů analýza MeSH
- Světová zdravotnická organizace MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- srovnávací studie MeSH
In 2006, levels of seven indicator polychlorinated biphenyl congeners (PCB28, 52, 101, 118, 138, 153, and 180) in blood serum of 202 blood donors residing for more than 2 years in five urban areas included in the Czech Human Biomonitoring project were measured by GC/MS/MS method. PCB congeners 138, 153 and 180 accounted for about 97% of the sum of the indicator congeners analyzed. Overall, the median and 95th percentile of the most abundant congener PCB 153 were 438 ng/g lipid and 1079 ng/g lipid, respectively. The highest median levels were found in Uherske Hradiste (669 ng/g lipid) and Ostrava (672 ng/g lipid in males compared to 341 ng/g lipid in females). Serum PCB concentrations were significantly associated with age, gender, place of residence and smoking habit, but not with body mass index and education. The results suggest the importance of PCB body burden in the Czech general population and the existence of hot spots.
- MeSH
- dospělí MeSH
- financování organizované MeSH
- index tělesné hmotnosti MeSH
- látky znečišťující vzduch krev MeSH
- lidé středního věku MeSH
- lidé MeSH
- mladiství MeSH
- plynová chromatografie s hmotnostně spektrometrickou detekcí MeSH
- polychlorované bifenyly krev MeSH
- senioři MeSH
- sexuální faktory MeSH
- věkové faktory MeSH
- vystavení vlivu životního prostředí analýza MeSH
- zeměpis MeSH
- Check Tag
- dospělí MeSH
- lidé středního věku MeSH
- lidé MeSH
- mladiství MeSH
- mužské pohlaví MeSH
- senioři MeSH
- ženské pohlaví MeSH
- Geografické názvy
- Česká republika MeSH
Abnormalities in cancer metabolism represent potential targets for cancer therapy. We have recently identified a natural compound Quambalarine B (QB), which inhibits proliferation of several leukemic cell lines followed by cell death. We have predicted ubiquinone binding sites of mitochondrial respiratory complexes as potential molecular targets of QB in leukemia cells. Hence, we tracked the effect of QB on leukemia metabolism by applying several omics and biochemical techniques. We have confirmed the inhibition of respiratory complexes by QB and found an increase in the intracellular AMP levels together with respiratory substrates. Inhibition of mitochondrial respiration by QB triggered reprogramming of leukemic cell metabolism involving disproportions in glycolytic flux, inhibition of proteins O-glycosylation, stimulation of glycine synthesis pathway, and pyruvate kinase activity, followed by an increase in pyruvate and a decrease in lactate levels. Inhibition of mitochondrial complex I by QB suppressed folate metabolism as determined by a decrease in formate production. We have also observed an increase in cellular levels of several amino acids except for aspartate, indicating the dependence of Jurkat (T-ALL) cells on aspartate synthesis. These results indicate blockade of mitochondrial complex I and II activity by QB and reduction in aspartate and folate metabolism as therapeutic targets in T-ALL cells. Anti-cancer activity of QB was also confirmed during in vivo studies, suggesting the therapeutic potential of this natural compound.
- Publikační typ
- časopisecké články MeSH
N-Benzyl-2-nitrobenzenesulfonamides underwent base-mediated intramolecular arylation at the benzyl sp(3) carbon to yield benzhydrylamines. The presence of electron withdrawing groups on the aromatic ring of the benzyl group was required to facilitate the C-arylation. Unsymmetrically substituted benzhydrylamines are advanced intermediates toward nitrogenous heterocycles, as exemplified in the syntheses of indazole oxides and quinazolines.
- MeSH
- alkoholy chemie MeSH
- benzhydrylové sloučeniny chemie MeSH
- benzylové sloučeniny chemie MeSH
- chinazoliny chemická syntéza MeSH
- heterocyklické sloučeniny chemická syntéza MeSH
- indazoly chemická syntéza MeSH
- indikátory a reagencie MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
6-Chloropurines substituted at the position 9 with variously modified bicyclic skeletons represent promising antiviral and anticancer agents. This work aimed to investigate the transport mechanisms of 9-[(1R*,2R*,4S*)-bicyclo[2.2.1]hept-2-yl]-6-chloro-9H-purine (9-norbornyl-6-chloropurine, NCP) and their relationship to the metabolism and biological activity of the compound. Transport experiments were conducted in CCRF-CEM cells using radiolabeled compound ([(3)H]NCP). The pattern of the intracellular uptake of [(3)H]NCP in CCRF-CEM cells pointed to a combination of passive and facilitated diffusion as prevailing transport mechanisms. NCP intracellular metabolism was found to enhance its uptake by modifying NCP concentration gradient. The transport kinetics reached steady state under the conditions of MRP and MDR proteins blockade, indicating that NCP is a substrate for these efflux pumps. Their inhibition also increased the cytotoxicity of NCP. Our findings suggest that the novel nucleoside analog NCP has potential to become a new orally available antileukemic agent due to its rapid membrane permeation.
- MeSH
- ABC transportéry antagonisté a inhibitory genetika metabolismus MeSH
- biologický transport MeSH
- buthionin sulfoximin farmakologie MeSH
- chinoliny farmakologie MeSH
- dibenzocyklohepteny farmakologie MeSH
- exprese genu MeSH
- kinetika MeSH
- kyselina ethakrynová farmakologie MeSH
- lidé MeSH
- nádorové buněčné linie MeSH
- permeabilita buněčné membrány účinky léků MeSH
- propionáty farmakologie MeSH
- protinádorové látky chemická syntéza metabolismus farmakologie MeSH
- puriny chemická syntéza metabolismus farmakologie MeSH
- T-lymfocyty účinky léků metabolismus patologie MeSH
- tritium MeSH
- usnadněná difuze MeSH
- viabilita buněk účinky léků MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
AIMS: TheAIM of our study is to present a novel approach for preparing a compound heterozygous reference material (hetRM) using gene synthesis technology with inverted insertion of wild-type and mutant fragments into a single cloning vector. Factor II (G20210A) and Factor V (G1691A Leiden) gene mutations were used as an experimental model. METHODS: During the gene synthesis, DNA fragments were aligned in the following order: G1691 FV wild-type forward strain, G20210 FII wild-type forward strain, 1691A FV mutant reverse strain, 20210A FII mutant reverse strain. The complete chain was inserted into a pIDT SMART cloning vector and amplified in an E. coli competent strain. For assessing hetRM characteristics and commutability, we used real-time PCR with subsequent melting curve analysis, real-time PCR with hydrolysis probes, allele-specific amplification, reverse hybridization, and dideoxynucleotide DNA sequencing. RESULT: All five methods yielded concordant results of DNA analysis of the hetRM. Differences in real-time PCR cycle threshold values after six-months of storage at -80 °C were not statistically significant from those obtained from freshly prepared hetRM aliquots, which is a good indication of their stability. CONCLUSION: By applying the procedures of gene synthesis and cloning technology, we prepared and verified a model genetic reference material for FII G20210A and FV G1691A testing with a compound heterozygous genotype. The hetRM was stable, commutable, and available in large quantities and in a wide concentration range.
- MeSH
- Escherichia coli MeSH
- faktor V genetika MeSH
- genetické techniky * MeSH
- klonování organismů metody MeSH
- kvantitativní polymerázová řetězová reakce MeSH
- mutace MeSH
- protrombin genetika MeSH
- sekvenční analýza DNA MeSH
- trombofilie genetika MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
