Selective enumeration
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Mycobacterium avium subsp. paratuberculosis (MAP) represents a slow-growing bacterium causing paratuberculosis, especially in domestic and wild ruminants. Until recently, the assessment of MAP viability relied mainly on cultivation, which is very time consuming and is unable to detect viable but non-culturable cells. Subsequently, viability PCR, a method combining sample treatment with the DNA-modifying agent ethidium monoazide (EMA) or propidium monoazide (PMA) and quantitative PCR (qPCR), was developed, enabling the selective detection of MAP cells with an intact cell membrane. However, this technology requires a laborious procedure involving the need to work in the dark and on ice. In our study, a method based on a combination of platinum compound treatment and qPCR, which does not require such a demanding procedure, was investigated to determine mycobacterial cell viability. The conditions of platinum compound treatment were optimized for the fast-growing mycobacterium M. smegmatis using live and heat-killed cells. The optimal conditions consisting of a single treatment with 100 μM cis-dichlorodiammine platinum(II) for 60 min at 5°C resulted in a difference in quantification cycle (Cq) values between live and dead membrane-compromised mycobacterial cells of about 6 Cq corresponding to about 2 log10 units. This optimized viability assay was eventually applied to MAP cells and demonstrated a better ability to distinguish between live and heat-killed mycobacteria as compared to PMA. The viability assay combining the Pt treatment with qPCR thereby proved to be a promising method for the enumeration of viable MAP cells in foodstuffs, environmental, and clinical samples which could replace the time-consuming cultivation or laborious procedures required when using PMA.
- Publikační typ
- časopisecké články MeSH
Objectives. The aim of the study was to highlight the specific features in the “gentleman and dog” drawings of children with hearing impairment who experience problems with verbalization. Sample and setting. The primary sample was 53 preschool children with hearing impairment. The design of the research was mixed. The drawings were qualitatively analysed with an enumeration of character frequency. The hypotheses were verified by a two-factor analysis and a two-sample T-test. Hypotheses. H1 There is no relation between the level of intelligence and the drawing. H2 There is no relation between the drawing and the sex of the child. H3 There is no relation between the drawing and the age of the child. H3 There is no relation between the drawing and the hearing impairment of the parents. Statistical analysis. There was a correlation between the results in the IQ test and the raw scores of the gentleman drawing at the level of 0.05 and the IQ test results and the raw scores of the dog drawing at level 0.01. The relationship between the sex, age, and level of the gentleman drawing has not been established. In the case of dog drawing, a statistically significant effect on the significance level of 0.05 only for sex (F (1, 48) = 6.15, p = 0.02) was demonstrated. In the event of the influence of the hearing impairment of parents on the child drawing, a statistically significant relationship was not supported. Results. Drawings of “gentleman and dog” of children with hearing impairment show signs of a lower developmental level by one to two years compared to hearing peers. Limits of the study. From the point of view of statistical processing requirements, the number of respondents may be considered to be limiting, but this is 80% of the selected population.
Phage therapy could offer a safe and effective alternative to antibiotic treatment of infections caused by Gram-positive bacterium Staphylococcus aureus that have emerged as a significant threat in hospital and community environment and is attracting growing interest among clinicians. The legislation process of approving the phage therapeutics by pharmaceutical authorities requires rapid analytical techniques for assessment of phage activity. Here, we present a three-step method for on-line monitoring the phage effect on bacterial cells dynamically adhered from microliter volumes of high conductivity matrix onto the inner surface of fused silica capillary with a part etched with supercritical water. Phage K1/420 particles of the Kayvirus genus generated by propagation on the host S. aureus cells together with the uninfected cells were concentrated, separated and detected using capillary electrophoretic methods. The phage interactions with selected S. aureus strains exhibiting differences in phage susceptibility were compared. The method allowed determination of the phage burst size and time of phage latent period in analyzed strains. Apart from enumeration of bacteriophages by the plaque assays, the proposed method is suitable for phage activity testing.
- MeSH
- antibakteriální látky MeSH
- bakteriofágy * MeSH
- lidé MeSH
- oxid křemičitý MeSH
- stafylokokové infekce * MeSH
- Staphylococcus aureus MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
Developmental responses to auxin are regulated by facilitated uptake and efflux, but detailed molecular understanding of the carrier proteins is incomplete. We have used pharmacological tools to explore the chemical space that defines substrate preferences for the auxin uptake carrier AUX1. Total and partial loss-of-function aux1 mutants were assessed against wild-type for dose-dependent resistance to a range of auxins and analogues. We then developed an auxin accumulation assay with associated mathematical modelling to enumerate accurate IC50 values for a small library of auxin analogues. The structure activity relationship data were analysed using molecular field analyses to create a pharmacophoric atlas of AUX1 substrates. The uptake carrier exhibits a very high level of selectivity towards small substrates including the natural indole-3-acetic acid, and the synthetic auxin 2,4-dichlorophenoxyacetic acid. No AUX1 activity was observed for herbicides based on benzoic acid (dicamba), pyridinyloxyacetic acid (triclopyr) or the 6-arylpicolinates (halauxifen), and very low affinity was found for picolinic acid-based auxins (picloram) and quinolinecarboxylic acids (quinclorac). The atlas demonstrates why some widely used auxin herbicides are not, or are very poor substrates. We list molecular descriptors for AUX1 substrates and discuss our findings in terms of herbicide resistance management.
- MeSH
- Arabidopsis metabolismus MeSH
- biologické modely MeSH
- biotest MeSH
- herbicidy metabolismus MeSH
- indoly metabolismus MeSH
- inhibiční koncentrace 50 MeSH
- kořeny rostlin růst a vývoj MeSH
- kyselina 2,4-dichlorfenoxyoctová metabolismus MeSH
- kyseliny indoloctové metabolismus MeSH
- mutace genetika MeSH
- proteiny huseníčku metabolismus MeSH
- semenáček růst a vývoj MeSH
- substrátová specifita MeSH
- tabák cytologie MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
... Messora -- S41 Flow Cytometry: Evolution of Microbiological -- Methods for Probiotics Enumeration -- ... ... , please contact the Ovid Regional Sales Office near you or visit www.ovid.com/site/index.jsp and select ... ... Selected articles are abstracted in Modern Medicine. ...
Journal of clinical gastroenterology, ISSN 0192-0790 Volume 52, Supplement 1, November/December 2018
115 stran : ilustrace ; 28 cm
- MeSH
- léčivé rostliny MeSH
- nutriční vědy MeSH
- potraviny MeSH
- potravní doplňky MeSH
- prebiotika MeSH
- probiotika MeSH
- střevní mikroflóra MeSH
- Publikační typ
- kongresy MeSH
- sborníky MeSH
- zprávy MeSH
- Konspekt
- Patologie. Klinická medicína
- NLK Obory
- gastroenterologie
- nutriční terapie, dietoterapie a výživa
BACKGROUND: We present a series of papillary renal cell carcinomas (PRCC) reminiscent of so-called "oncocytic variant of papillary renal cell carcinoma" (OPRCC), included in the 2016 WHO classification as a potential type 3 PRCC. OPRCC is a poorly understood entity, cytologically characterized by oncocytic cells with non-overlapping low grade nuclei. OPRCC is not genotypically distinct and the studies concerning this variant have shown an inconsistent genetic profile. The tumors presented herein demonstrated predominantly papillary/tubulopapillary architecture and differed from OPRCC by pseudostratification and grade 2-3 nuclei (Fuhrman/ISUP). Because there is a morphologic overlap between renal oncocytoma (RO) and PRCC in the cases included in this study, the most frequently affected chromosomes in RO and PRCC were analyzed. MATERIALS AND METHODS: 147 PRCC composed of oncocytic cells were retrieved from our registry in order to select a group of morphologically uniform tumors. 10 cases with predominantly papillary, tubulopapillary or solid architectural patterns were identified. For immunohistochemical analysis, the following antibodies were used: vimentin, antimitochondrial antigene (MIA), AMACR, PAX8, CK7, CK20, AE1-3, CAM5.2, OSCAR, Cathepsin K, HMB45, SDHB, CD10, and CD117. Enumeration changes of locus 1p36, chromosomes 7, 14, 17, X, Y and rearrangement of CCND1 were examined by FISH. For further study, only tumors showing karyotype similar to that of RO were selected. The tumors exhibiting either trisomy of chromosomes 7, 17 or gain of Y, thus abnormalities characteristic for PRCC, were excluded. RESULTS: There were 5 males and 5 females, with patient age ranging from 56 to 79 years (mean 66.8 years). The tumor size ranged from 2 to 10 cm (mean 5.1 cm). Follow-up was available for 8/10 patients (mean 5.2 years); one patient died of the disease, while 7 of 8 are alive and well. Immunohistochemically, all cases were reactive for AMACR, vimentin, PAX8, OSCAR, CAM5.2, and MIA. SDHB was retained in all cases. 9/10 cases were positive for CD10, 7/10 cases reacted with CK7, 4/10 with Cathepsin K, and 2/10 with AE1-3. None of the cases were positive for CD117, HMB45 and CK20. All 10 cases were analyzable by FISH and showed chromosomal abnormalities similar to that usually seen in RO (i.e. loss of 1p36 gene loci, loss of chromosome Y, rearrangement of CCND1 and numerical changes of chromosome 14). CONCLUSIONS: We analyzed a series of renal tumors combining the features of PRCC/OPRCC and RO, that included pseudostratification and mostly high grade oncocytic cells lining papillary/tubulopapillary structures, karyotype characterized by loss of 1p36, loss of chromosome Y, rearrangement of CCND1 gene and numerical changes of chromosome 14. Despite the chromosomal numerical abnormalities typical of RO, we classified these tumors as part of the spectrum of PRCC because of their predominant papillary/tubulopapillary architecture, immunoprofile that included reactivity for AMACR, vimentin and lack of reactivity for CD117, all of which is incompatible with the diagnosis of RO. This study expands the morphological spectrum of PRCC by adding a cohort of diagnostically challenging cases, which may be potentially aggressive.
- MeSH
- chromozomální aberace MeSH
- diferenciální diagnóza MeSH
- karcinom z renálních buněk genetika metabolismus patologie MeSH
- lidé středního věku MeSH
- lidé MeSH
- nádorové biomarkery genetika metabolismus MeSH
- nádory ledvin genetika metabolismus patologie MeSH
- oxyfilní adenom genetika metabolismus patologie MeSH
- papilární karcinom genetika metabolismus patologie MeSH
- senioři MeSH
- Check Tag
- lidé středního věku MeSH
- lidé MeSH
- mužské pohlaví MeSH
- senioři MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
Eligibility to anti-HER2 therapy for breast tumors strictly depends on demonstrating HER2 overexpression (by immunohistochemistry) or HER2 gene amplification by in situ hybridization (ISH), usually defined by the ratio of HER2 gene to chromosome 17 centromere (CEP17) copies. However, the CEP17 copy number increase (CNI) has been proven responsible for misleading HER2 FISH results and recent small cohort studies suggest that chromosome 17 polysomy is actually very rare. Here we investigated by FISH the frequency of true chromosome 17 polysomy in a consecutive cohort of 5,477 invasive breast cancer patients. We evaluated and selected the LSI 17p11.2 probe for chromosome 17 enumeration on a training cohort of 67 breast cancer samples (CEP17 ≥ 2.5). LSI 17p11.2 was used in the 297/5,477 patients from the validation cohort displaying CEP17 CNI (CEP17 ≥ 3.0). Using HER2/17p11.2 scoring criteria, 37.3%/1.5% patients initially classified as equivocal/non-amplified were reclassified as amplified. For a more accurate assessment of chromosome 17 and ploidy in the samples, we tested six markers located on chromosome 17 and centromeric regions of chromosome 8 (CEP8) and 11 (CEP11) in 67 patients with CEP17 and LSI 17p11.2 CNI. True polysomy (hyperdiploidy) according to these markers was found in 0.48% of cases (24/5,020). CEP8 and CEP11 CNI (≥3.0) was more frequent in the hyperdiploid than CEP17 non-polysomic group (55.6% vs. 6.1% and 25% vs. 2.3%, respectively). Our results suggest that chromosome 17 polysomy is a rare event found in <1% breast cancer cases and that polysomy of other chromosomes frequently occurs with chromosome 17 polysomy.
Přehledné sdělení týkající se chronické stabilní anginy pectoris, její definice, praktického postupu vyšetření, volby jednotlivých druhů zátěžových testů a stratifikace rizika nemocných. Jsou uvedeny principy farmakologické léčby, která sleduje 2 hlavní cíle: odstranění symptomů (antiischemická léčba) a zlepšení prognózy (prevence kardiovaskulárních příhod) a jsou přehledně vyjmenovány základní indikace ke koronarografii a revaskularizaci myokardu.
Review article dealing with chronic stable angina pectoris, its definition, practical examinations, selection of different types of stress tests and patient risk stratification. Principles of pharmacological treatment, which pursues two main objectives: the elimination of symptoms (ischemic treatment) and improved prognosis (prevention of cardiovascular events), are clearly enumerated as well as basic indications for coronary angiography and revascularization.
- MeSH
- angina pectoris * diagnóza patofyziologie terapie MeSH
- beta blokátory terapeutické užití MeSH
- blokátory kalciových kanálů terapeutické užití MeSH
- elektrokardiografie MeSH
- hodnocení rizik MeSH
- inhibitory agregace trombocytů terapeutické užití MeSH
- koronární angiografie MeSH
- lidé MeSH
- nitroglycerin terapeutické užití MeSH
- revaskularizace myokardu MeSH
- rizikové faktory MeSH
- zátěžový test metody MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- přehledy MeSH
An international standard already exists for the selective enumeration of bifidobacteria in milk products. This standard uses Transgalactosylated oligosaccharides (TOS) propionate agar supplemented with mupirocin. However, no such standard method has been described for the selective enumeration of bifidobacteria in probiotic supplements, where the presence of bifidobacteria is much more variable than in milk products. Therefore, we enumerated bifidobacteria by colony count technique in 13 probiotic supplements using three media supplemented with mupirocin (Mup; 100mg/l): TOS, Bifidobacteria selective medium (BSM) and modified Wilkins-Chalgren anaerobe agar with soya peptone (WSP). Moreover, the potential growth of bifidobacterial strains often used in probiotic products was performed in these media. All 13 products contained members of the genus Bifidobacterium, and tested mupirocin media were found to be fully selective for bifidobacteria. However, the type strain Bifidobacterium bifidum DSM 20456 and collection strain B. bifidum DSM 20239 showed statistically significant lower counts on TOS Mup media, compared to BSM Mup and WSP Mup media. Therefore, the TOS Mup medium recommended by the ISO standard cannot be regarded as a fully selective and suitable medium for the genus Bifidobacterium. In contrast, the BSM Mup and WSP Mup media supported the growth of all bifidobacterial species.
- MeSH
- antibakteriální látky metabolismus MeSH
- bakteriální nálož metody MeSH
- Bifidobacterium účinky léků růst a vývoj MeSH
- kultivační média chemie MeSH
- mupirocin metabolismus MeSH
- počet mikrobiálních kolonií metody MeSH
- probiotika analýza MeSH
- Publikační typ
- časopisecké články MeSH
- hodnotící studie MeSH
- práce podpořená grantem MeSH
- srovnávací studie MeSH
Cíl: Cílem výzkumu bylo zmapovat specifika komunikace o smrti s dítětem v rodině se zdravými členy. Design: Kvalitativní výzkum. Metodika: Pro počáteční orientaci v dosud neprozkoumané oblasti byla zvolena kvalitativní metodologie. Metoda polostrukturovaného rozhovoru byla zacílena na matky předškolních dětí od 5 do 6 let, v jejichž nukleární rodině se nevyskytuje umírající člen. Kombinací nepravděpodobnostních metod výběru vzorku bylo vybráno 14 matek. K analýze dat se využilo metody vytváření trsů, metody prostého výčtu a metody zachycení gestaltů. Výsledky: Získaná data jsou deskripcí rodinného rozhovoru o smrti. Popisují osobu matky a dítěte, stejně tak i okolnosti, ve kterých tématická debata probíhá. Výsledky poukazují na intenzivní zájem dítěte o téma smrti, který je provázen většinovou tendencí matek informace dítěti zprostředkovat. Závěr: Téma smrti se v rodinném rozhovoru s předškolním dítětem ukázalo jako aktuální. Přesto, že je v současné společnosti smrt stále percipována negativně, členové rodiny by měli být na možnost otevření tématu před dětmi připraveni.
Aim: The aim of the research was to map the specifications of communication of death with a child in the family of healthy members. Design: Qualitative research. Methods: A qualitative methodology was selected for initial orientation in a hitherto unexplored area. The method of semi-structured interview was aimed at mothers of preschool children aged between five and six, within their immediate family with no terminally ill family members. 14 mothers were selected by combining improbability sampling methods. A cluster method, simple enumeration method and method of searching for patterns were used to analyse the data. Results: The obtained data describe family discussion about death. They describe mother and child as well as the circumstances in which thematic discussion takes place. The results indicate the strength of children ́s interest in the topic of death, which is accompanied by the tendency of the majority of mothers to convey such information to the child. Conclusion: The theme of death in family discussion with preschool children has proved topical. Despite the fact that contemporary society continues to perceive death negatively, family members should be prepared to explore the topic with their children.
