non-invasive sampling
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The discovery of presence of fetal non cell DNA in mother's blood and possibility of their measurement in 1997 (Lo ,USA) opened perspectives of safe prenatal screening of aneuploidies. Invasive diagnostics procedures like chorionic villus sampling and amniocentesis which have been widely implemented since 70's and 80's present risk of pregnancy loss in amount 1:100 or 1: 50. This approach of invasive and risky prenatal screening in time of demographic disaster in Europe is no longer unsustainable. NIPT on the contrary represents method which is completely safe and achieves the same accuracy regarding screening of trisomy 21 (Down syndrome). Results Our centre has performed about 100 NIPT's (MaterniT21, Harmony, Panorama Prenascan) since 2012. NIPT is offered only to those pregnant women who have low or intermediate risk of aneuploidies after first trimester screening. High percentage of pregnant after assisted reproduction procedures are among those tested by NIPT. The NIPT is a good choice for pregnant women after IVF who are already in advanced age in comparison to normal population. Invasive procedures like CVS or amniocentesis are recommended in case of high risk of aneuploidies ( calculation risk 1:10 resp. 1:50) or ultrasound abnormal findings. We have experience that running NIPT technology is very easy, and has been accepted our patients. Conclusions Our experience with NIPT is very positive. Main advantage of this new technology is safety and we are sure that NIPT is ideal for using in gynaecological practise. The first trimester ultrasound skills and NIPT technology allows gynaecologist to detect majority of aneupliodies and congenital anomalies of fetus.
- MeSH
- amniocentéza škodlivé účinky MeSH
- aneuploidie MeSH
- druhý trimestr těhotenství krev MeSH
- genetické nemoci vrozené krev ultrasonografie MeSH
- genetické poradenství metody normy MeSH
- genetické testování * metody MeSH
- lidé MeSH
- prenatální diagnóza * metody MeSH
- první trimestr těhotenství * krev MeSH
- těhotenství MeSH
- ultrasonografie prenatální metody přístrojové vybavení MeSH
- Check Tag
- lidé MeSH
- těhotenství MeSH
- ženské pohlaví MeSH
Where microbes colonizing skin surface may help maintain organism homeostasis, those that invade living skin layers cause disease. In bats, white-nose syndrome is a fungal skin infection that affects animals during hibernation and may lead to mortality in severe cases. Here, we inferred the amount of fungus that had invaded skin tissue of diseased animals. We used simulations to estimate the unobserved disease severity in a non-lethal wing punch biopsy and to relate the simulated pathology to the measured fungal load in paired biopsies. We found that a single white-nose syndrome skin lesion packed with spores and hyphae of the causative agent, Pseudogymnoascus destructans, contains 48.93 pg of the pathogen DNA, which amounts to about 1560 P destructans genomes in one skin lesion. Relating the information to the known UV fluorescence in Nearctic and Palearctic bats shows that Nearctic bats carry about 1.7 µg of fungal DNA per cm2, whereas Palearctic bats have 0.04 µg cm-2 of P. destructans DNA. With the information on the fungal load that had invaded the host skin, the researchers can now calculate disease severity as a function of invasive fungal growth using non-destructive UV light transillumination of each bat's wing membranes. Our results will enable and promote thorough disease severity assessment in protected bat species without the need for extensive animal and laboratory labor sacrifices.
- MeSH
- Ascomycota * metabolismus patogenita MeSH
- Chiroptera mikrobiologie MeSH
- dermatomykózy * mikrobiologie prevence a kontrola terapie veterinární MeSH
- hibernace * MeSH
- křídla zvířecí mikrobiologie MeSH
- kůže mikrobiologie MeSH
- ultrafialové záření * MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Článek se zabývá popisem a kritickým hodnocením vývoje přístrojové techniky pro semiinvazivní a neinvazivní stanovení glukózy v letech 1996 až 2001. Jsou uvedeny techniky odběru vzorku a vysvětleny principy optických a elektrochemických měřících metod používaných v prototypech přístrojů. Zatím nelze doporučit neinvazivní nebo minimálně invazivní metody analýzy glukózy jako náhradu monitorování krevní glukózy pacientem či biochemickou laboratoří.
The paper deals with description and critical evaluation of the instrumentation trends towards semiinvasive and non-invasive glucose determination from 1996 to 2001. The sample collection modes are mentioned and the principles of optical and electrochemical measuring methods used in prototypes of instruments are explained. The use of non-invasive or minimal invasive glucose analysis methods instead of blood glucose monitoring by the patient or the biochemical lab cannot be recommended yet.
In accordance with the 3 Rs principle (to replace, reduce and refine) animal models in biomedical research, we have developed and applied a new approach for sampling and analyzing hair follicles in various experimental settings. This involves use of a convenient device for non-invasive collection of hair follicles and processing methods that provide sufficient amounts of biological material to replace stressful and painful biopsies. Moreover, the main components of hair follicles are live cells of epithelial origin, which are highly relevant for most types of malignant tumors, so they provide opportunities for studying aging-related pathologies including cancer. Here, we report the successful use of the method to obtain mouse hair follicular cells for genotyping, quantitative PCR, and quantitative immunofluorescence. We present proof of concept data demonstrating its utility for routine genotyping and monitoring changes in quality and expression levels of selected proteins in mice after gamma irradiation and during natural or experimentally induced aging. We also performed pilot translation of animal experiments to human hair follicles irradiated ex vivo. Our results highlight the value of hair follicles as biological material for convenient in vivo sampling and processing in both translational research and routine applications, with a broad range of ethical and logistic advantages over currently used biopsy-based approaches.
- MeSH
- fluorescenční protilátková technika MeSH
- genotypizační techniky MeSH
- myši inbrední C57BL MeSH
- myši MeSH
- ocas patologie MeSH
- poškození DNA * účinky záření MeSH
- stárnutí patologie fyziologie MeSH
- vlasový folikul anatomie a histologie metabolismus fyziologie účinky záření MeSH
- zvířata MeSH
- Check Tag
- mužské pohlaví MeSH
- myši MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
V této prospektivní studii shrnujeme naše první zkušenosti s neinvazivní prenatální RH genotypizací plodu u aloimunizovaných těhotenství s využitím PCR v reálném čase, specifických primerů a TaqMan sond navržených pro RHD a RHCE geny. Celkem bylo testováno 22 aloimunizovaných těhotných žen (15 anti-D, 5 anti-D+C, 2 anti-E) v rozmezí 11. až 33. týdne gravidity. Výsledky neinvazivní prenatální RHD a RHCE genotypizace plodu provedené na DNA extrahované z periferní krve matek jsme korelovali s výsledky imunohematologické analýzy pupečníkové krve. Výsledky RHD a RHCE genotypizace plodu byly ve shodě s vyšetřením pupečníku u všech vyšetřených aloimunizovaných těhotných žen. Neinvazivní prenatální RHD a RHCE genotypizace plodu umožňuje identifikaci plodů v riziku hemolytického onemocnění. Detekce negativních plodů v současné graviditě může vyloučit potřebu invazivního prenatálního vyšetření.
In this prospective study, we described our first experience with non-invasive foetal RH genotyping in alloimmunised pregnancies by analysis of DNA extracted from maternal plasma samples by using real-time PCR and primers and probes targeted toward RHD and RHCE genes. We analysed 22 alloimmunised pregnant women (15 anti-D, 5 anti-D+C, 2 anti-E) within 11th and 33rd week of pregnancy and correlated the results with serological analysis of cord blood. Non-invasive prenatal foetal RHD and RHCE genotyping analysis of maternal plasma samples was in complete concordance with the analysis of cord blood in all alloimmunised pregnancies. Non-invasive foetal RHD and RHCE genotyping enables the identification of foetuses at risk of haemolytic disease of the newborn. An identification of negative foetuses may exclude the demand of invasive prenatal procedures.
- MeSH
- antikoagulancia farmakologie klasifikace terapeutické užití MeSH
- Publikační typ
- přehledy MeSH
- směrnice pro lékařskou praxi MeSH
BACKGROUND: In endemic areas of zoonotic leishmaniosis caused by L. infantum, early detection of Leishmania infection in dogs is essential to control the dissemination of the parasite to humans. The aim of this study was to evaluate the serological and/or molecular diagnostic performance of minimally and non-invasive samples (conjunctiva cells (CS) and peripheral blood (PB)) for monitoring Leishmania infection/exposure to Phlebotomus perniciosus salivary antigens in dogs at the beginning and the end of sand fly seasonal activity (May and October, respectively) and to assess associated risks factors. METHODS: A total of 208 sheltered dogs from endemic areas of leishmaniosis were screened. Leishmania DNA detection in PB on filter paper and CS was performed by nested-PCR (nPCR), while the detection of anti-Leishmania antibodies was performed using IFAT and ELISA. The exposure to P. perniciosus salivary antigens (SGH, rSP01 and rSP03B + rSP01) was measured by ELISA. RESULTS: Ninety-seven (46.6%) and 116 (55.8%) of the 208 dogs were positive to Leishmania antibodies or DNA by at least one test at the beginning and end of the sand fly season, respectively. IFAT and ELISA presented a substantial agreement in the serodiagnosis of leishmaniosis. Discrepant PB nPCR results were obtained between sampling points. Leishmania DNA was detected in CS of 72 dogs at the end of the phlebotomine season. The presence of antibodies to the parasite measured by ELISA was significantly higher in dogs presenting clinical signs compatible with leishmaniosis at both sampling points. Phlebotomus perniciosus salivary antibodies were detected in 179 (86.1%) and 198 (95.2%) of the screened dogs at the beginning and end of the phlebotomine season, respectively. CONCLUSIONS: The association between ELISA positivity and clinical signs suggests its usefulness to confirm a clinical suspicion. CS nPCR seems to be an effective and non-invasive method for assessing early exposure to the parasite. PB nPCR should not be used as the sole diagnostic tool to monitor Leishmania infection. The correlation between the levels of antibodies to P. perniciosus saliva and Leishmania antibodies suggests the use of a humoral response to sand fly salivary antigens as biomarkers of L. infantum infection.
- MeSH
- antigeny protozoální imunologie MeSH
- endemické nemoci prevence a kontrola MeSH
- hmyz - vektory parazitologie MeSH
- hmyzí proteiny imunologie MeSH
- imunoglobulin G krev MeSH
- konjunktiva cytologie parazitologie MeSH
- kousnutí a bodnutí hmyzem MeSH
- Leishmania infantum izolace a purifikace MeSH
- leishmanióza krev imunologie veterinární MeSH
- nemoci psů parazitologie prevence a kontrola přenos MeSH
- Phlebotomus parazitologie MeSH
- protilátky protozoální krev MeSH
- protozoální proteiny imunologie MeSH
- psi MeSH
- rizikové faktory MeSH
- sérologické testy MeSH
- slinné proteiny a peptidy imunologie MeSH
- zvířata MeSH
- Check Tag
- psi MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
No study has systematically compared the suitability of DNA methylation (DNAme) profiles in non-invasive samples for the detection of breast cancer (BC). We assess non-tumour DNAme in 1,100 cervical, buccal, and blood samples from BC cases and controls and find that cervical samples exhibit the largest nuber of differentially methylated sites, followed by buccal samples. No sites were significant in blood after FDR adjustment. Deriving DNAme-based classifiers for BC detection in each sample type (WID-buccal-, cervical-, or blood-BC), we achieve validation AUCs of 0.75, 0.66, and 0.51, respectively. Buccal and cervical BC-associated DNAme alterations distinguish between BC cases and controls in both surrogate and breast tissue (AUC > 0.88), yet individual sites and the directionality of methylation changes are not identical between these two sample types, and buccal sample DNAme aligns with breast methylation changes more closely. Pending additional validation, these insights may have the potential to improve non-invasive personalized BC prevention.
- MeSH
- dospělí MeSH
- epigeneze genetická * MeSH
- epigenomika * metody MeSH
- lidé středního věku MeSH
- lidé MeSH
- metylace DNA * MeSH
- nádory prsu * genetika diagnóza MeSH
- studie případů a kontrol MeSH
- ústní sliznice metabolismus MeSH
- Check Tag
- dospělí MeSH
- lidé středního věku MeSH
- lidé MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
INTRODUCTION: The methods for diagnosing compartment syndrome non-invasively remain under debate. Bioimpedance measurements offer a promising avenue in clinical practice, detecting subtle changes in organ impedance due to volume shifts. This study explores bioimpedance measurement as a novel, painless method for diagnosing compartment syndrome, potentially enabling continuous monitoring. OBJECTIVE: This work aims to develop a prototype device for non-invasive diagnosis of compartment syndrome based on bioimpedance changes and assess initial results through in vitro experiments on inanimate biological material. We assume a change in the bioimpedance value after the application of physiological solution. MATERIALS AND METHODS: Between 2018 and 2022, a prototype device for diagnosing limb compartment syndrome was collaboratively developed with the Department of Cybernetics and Biomedical Engineering at the Technical University of Ostrava, Czech Republic. This device operates by comparing bioimpedance between two compartments, one of which is pathologically affected (experiencing compartment syndrome). The Bioimpedance Analyzer for Compartment Syndrome (BACS) has been utilized to conduct measurements on inanimate biological material in laboratory settings. Two samples of duck and chicken tissue, as well as piglets, were employed for these experiments. According to the size of sample was compartment syndrome simulated by injecting 20-120 mL saline into one limb (breast) while leaving the other as a control. Invasive intramuscular pressure measurements were conducted post-saline injection using a conventional device (Stryker). Changes in bioimpedance were evaluated following saline application. RESULTS: The non-invasive bioimpedance measurement instrument has been developed. It meets the safety requirements of European standard EN 60601-1. Measurement of accuracy showed minimal deviation for both channels (1.08% for the left channel and 1.84% for the right channel) when measuring on resistors. Ten measurements were conducted using the BACS prototype - two on chicken legs, two on duck breasts, two on duck legs, and four on piglets. Compartment syndrome simulation was achieved for all 10 measurements (IMP variance 31-45 mmHg). Following saline application, a notable decrease in bioimpedance was observed in the compartment simulating compartment syndrome (decrease by 12-78 Ω). CONCLUSION: Non-invasive methods could revolutionize limb compartment syndrome diagnosis, offering advantages such as non-invasiveness and continuous monitoring of compartment swelling.
- Publikační typ
- časopisecké články MeSH
Provedli jsme modifikaci a optimalizaci postupů pro izolaci extracelulární DNA z mateřské cirkulace pro neinvazivní určení pohlaví plodu u těhotenství s rizikem X-vázaných onemocnění u plodu a pro neinvazivní RHD a RHCE genotypizaci plodu u aloimunizovaných těhotenství s rizikem hemolytického onemocnění plodu. V této studii srovnáváme výsledky amplifikace SRY, RHD a RHCE genu na extracelulární DNA izolované z mateřské periferní krve pomocí QIAamp DSP Virus kitu a QIAamp DNA Blood Mini kitu. Výsledky našich analýz prokázaly, že QIAamp DSP Virus kit zvyšuje výtěžnost extracelulární fetální DNA přítomné v mateřské plazmě, což je klíčové zejména u amplifikace paternálně zděděných alel, které se liší od alel maternálních pouze v jednom nukleotidu. Doporučujeme proto provádět detekci paternální Rhc alely a RhE alely RHCE genu na extracelulární DNA izolované z mateřké plazmy pouze pomocí QIAamp DSP Virus kitu. Pro amplifikaci SRY a RHD genu dostačuje extracelulární DNA extrahovaná z mateřské plazmy prostřednictvím QIAamp DNA Blood Mini kitu. V případě sporných výsledků, tj. při nálezu pozdních amplifikačních cyklů (po 40. cyklu), doporučujeme rovněž provedení PCR analýzy na extracelulární DNA izolované z mateřské plazmy pomocí QIAamp DSP Virus kitu. Spolehlivá detekce plodů negativních na aglutinogeny, vůči nimž je matka aloimunizována, vylučuje riziko fetální erytroblastózy. Spolehlivá detekce plodů ženského pohlaví u těhotenství s rizikem X-vázaných onemocnění může vyloučit potřebu invazivního prenatálního vyšetření.
We carried out the modification and optimalisation of extracellular fetal DNA isolation from maternal circulation for non-invasive fetal sex determination in pregnancies at risk of X-linked disorders and for non-invasive fetal RHD and RHCE genotyping in alloimmunized pregnancies at risk of haemolytic disease of newborn. In this study, we compared the results of fetal SRY, RHD and RHCE genotyping by analysis of DNA extracted from maternal plasma samples by using QIAamp DSP Virus kit and QIAamp DNA Blood Mini kit. We showed that QIAamp DSP Virus kit enhanced the recovery of extracellular fetal DNA from maternal plasma that is crucial especially for the detection of those paternally-inherited alleles that differ from maternal alleles only in one nucleotide. Therefore we recommend to perform the detection of fetal Rhc allele and RhE allele of RHCE gene by analysis of extracellular DNA extracted from maternal plasma samples by using QIAamp DSP Virus kit only. We showed that the use of extracellular DNA extracted from maternal plasma samples by using QIAamp DNA Blood Mini kit is sufficient for non-invasive fetal SRY and RHD genotyping. In case of controversial results due to the late amplification (Ct ≥ 40) we recommend to perform real-time PCR analysis on extracellular DNA extracted from maternal plasma samples by using QIAamp DSP Virus kit. Reliable non-invasive detection of negative foetuses in alloimmunized pregnancies may exclude the risk of haemolytic disease of newborn. Reliable non-invasive detection of female foetuses in pregnancies at risk of X-linked disorders may exclude the performance of invasive prenatal diagnostic procedures.
- MeSH
- analýza určování pohlaví metody MeSH
- finanční podpora výzkumu jako téma MeSH
- genetické nemoci vázané na chromozom X prevence a kontrola MeSH
- hemolytická nemoc plodu a novorozence prevence a kontrola MeSH
- lidé MeSH
- odběr vzorku krve využití MeSH
- prenatální diagnóza metody MeSH
- reagenční diagnostické soupravy využití MeSH
- sekvenční analýza DNA MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- hodnotící studie MeSH
Kondenzát vydechovaného vzduchu je tekutina získána chlazením vzduchu při klidovém dýchání ústy. Takto získaný materiál obsahuje látky nacházející se v dolních dýchacích cestách podobně jako bronchoalveolární tekutina. Kondenzát obsahuje mnoho mediátorů a znaků probíhajícího zánětu sliznice dolních dýchacích cest, např. metabolity kyseliny arachidonové (leukotrieny, isoprostany, prostaglandiny), proteiny (cytokiny), vazoaktivní peptidy a řadu dalších, jako je peroxid vodíku, nitrity/nitráty, nitrotyrozin a další. Autoři uvádějí současné možnosti odběru, vyšetření i limity u chronických plicních nemocí, hlavně u asthma bronchiale a chronické obstrukční pulmonální nemoci (CHOPN), u kterých je možné vyšetřením kondenzátu vydechovaného vzduchu zánět detekovat ještě v bezpříznakovém období a sledovat jeho tíži.
Exhaled breath condensate is a liquid obtained by cooling the air at the calm mouth breathing. That way obtained material conta ins sub- stances located in the lower respiratory tract similar as bronchoalveolar fluid. The condensate contains many mediators and sig ns of inflammation of lower respiratory tract mucous such as metabolites of arachidonic acid (leukotrienes, isoprostanes, prostagland ins), proteins (cytokines), vasoactive peptides and many others such as hydrogen peroxide, nitrites/nitrates, nitrotyrozin and more. The au- thors present current possibilities of sampling, testing and limits at chronic pulmonary diseases, especially for asthma and CO PD, where is possible by examination of exhaled breath condensate detected inflammation even in asymptomatic period and observe their severity.
- MeSH
- bronchiální astma MeSH
- dechové testy * metody přístrojové vybavení MeSH
- diagnostické techniky dýchacího ústrojí MeSH
- dýchání MeSH
- ikosanoidy MeSH
- klinické zkoušky jako téma MeSH
- lidé MeSH
- mediátory zánětu MeSH
- nemoci dýchací soustavy * diagnóza MeSH
- nukleosidy MeSH
- nukleotidy MeSH
- odběr biologického vzorku MeSH
- oxid dusnatý MeSH
- proteomika MeSH
- puriny MeSH
- vydechnutí MeSH
- vzduch * analýza MeSH
- zánět MeSH
- Check Tag
- lidé MeSH
