PURPOSE: Preimplantation genetic testing for monogenic disorders (PGT-M) allows early diagnosis in embryos conceived in vitro. PGT-M helps to prevent known genetic disorders in affected families and ensures that pathogenic variants in the male or female partner are not passed on to offspring. The trend in genetic testing of embryos is to provide a comprehensive platform that enables robust and reliable testing for the causal pathogenic variant(s), as well as chromosomal abnormalities that commonly occur in embryos. In this study, we describe PGT protocol that allows direct mutation testing, haplotyping, and aneuploidy screening. METHODS: Described PGT protocol called OneGene PGT allows direct mutation testing, haplotyping, and aneuploidy screening using next-generation sequencing (NGS). Whole genome amplification product is combined with multiplex PCR used for SNP enrichment. Dedicated bioinformatic tool enables mapping, genotype calling, and haplotyping of informative SNP markers. A commercial software was used for aneuploidy calling. RESULTS: OneGenePGT has been implemented for seven of the most common monogenic disorders, representing approximately 30% of all PGT-M indications at our IVF centre. The technique has been thoroughly validated, focusing on direct pathogenic variant testing, haplotype identification, and chromosome abnormality detection. Validation results show full concordance with Sanger sequencing and karyomapping, which were used as reference methods. CONCLUSION: OneGene PGT is a comprehensive, robust, and cost-effective method that can be established for any gene of interest. The technique is particularly suitable for common monogenic diseases, which can be performed based on a universal laboratory protocol without the need for set-up or pre-testing.
- MeSH
- aneuploidie MeSH
- blastocysta patologie MeSH
- genetické testování metody MeSH
- lidé MeSH
- mutace genetika MeSH
- preimplantační diagnóza * metody MeSH
- těhotenství MeSH
- vysoce účinné nukleotidové sekvenování metody MeSH
- Check Tag
- lidé MeSH
- mužské pohlaví MeSH
- těhotenství MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
Pochopení vztahu mezi senzomotorickými proměnnými a exerkiny, které ovlivňují funkci mozku a kognici, nám umožňuje hlouběji porozumět biologickému procesu stárnutí. Hlavním cílem této studie bylo zjistit, jak silně jsou mozkový neurotrofický faktor (brain-derived neurotrophic factor, BDNF), irisin, svalová hmota a svalová síla asociovány s výsledky testů vybraných kognitivních funkcí u starších žen a jak dobře je predikují. Padesát sedm starších žen (průměrný věk 70,4 ± 4,1 roku) absolvovalo baterii neuropsychologických testů, měření izometrické dynamometrie a bioelektrické impedance. Hladiny v krevním séru sledovaných exerkinů byly stanoveny enzymatickým imunosorbentním testem (ELISA). Pro testování predikcí byly využity hierarchické vícenásobné regresní modely. Odhadli jsme, že rozptyl 46,1 % v krátkodobé paměti byl zapříčiněn hladinami BDNF v séru, přičemž druhým statisticky významným prediktorem byl věk (beta = –0,22; p = 0,030). Síla dolních končetin (lower limb strength, LLS) prokázala významnou prediktivní sílu jak u paměti – bezprostřední vybavení (beta = 0,39; p = 0,004), tak u paměti – oddálené vybavení (beta = 0,45; p = 0,001). Hladiny BDNF v séru byly významným prediktorem u oddáleného vybavení (beta = 0,29; p = 0,048). Přidání hladin BDNF do modelu prokázalo významné zvýšení jeho prediktivní síly o přibližně 5,6 % (p = 0,048) u paměti – oddálené vybavení. Index kosterní svalové hmoty (skeletal muscle index, SMI) a úroveň vzdělání byly významnými prediktory mentální flexibility. Byla zjištěna silná pozitivní asociace mezi hladinami BDNF, irisinem, svalovou silou a kognitivní funkcí, přičemž irisin a svalová síla jsou silnými prediktory hladin BDNF u starších žen. Studie byla realizována s podporou grantu Univerzity Karlovy – PRIMUS/19/HUM/012, Specifického vysokoškolského výzkumu SVV 260599, projektu COOPERATIO a Grantové agentury UK číslo grantu 268321. Korespondenční adresa: PhDr. Veronika Holá Katedra gymnastiky a úpolových sportů FTVS UK José Martího 269/31 162 52 Praha 6-Veleslavín e-mail: veronika.hola@ftvs.cuni.cz
Understanding the relationship between sensorimotor variables and exerkines related to brain function and cognition may help better understand biological ageing. The main aim of this study was to determine how strongly brain-derived neurotrophic factor (BDNF), irisin, muscle mass and muscle strength are associated and predict scores on selected cognitive domain tests in older women. Fifty seven older women (mean age 70.4 ± 4.1 years) underwent a battery of cognitive and psychological tests and measurements of isometric dynamometry and bioelectrical impedance. Serum exerkines levels were measured by enzyme-linked immunosorbent assay (ELISA). Hierarchical multiple regression models were used to test the predictions. We estimated that 46.1% of the variance in short-term memory was accounted for by serum BDNF levels, with age being the second statistically significant predictor (Beta = -0.22; p = 0.030). Lower limb strength (LLS) showed significant predictive power in both immediate (Beta = 0.39; p = 0.004) and delayed memory (Beta = 0.45; p = 0.001), serum BDNF levels were a significant predictor in delayed memory (Beta = 0.29; p = 0.048). Adding serum BDNF levels to the model showed a significant increase in predictive power of approximately 5.6% (p = 0.048) in delayed memory. Skeletal muscle index (SMI) and education level were significant predictors of mental flexibility. A strong positive association between BDNF levels, irisin, muscle strength, and cognitive function was found, with irisin and muscle strength being strong predictors of BDNF levels in older women.
- Klíčová slova
- irisin,
- MeSH
- fibronektinová doména typu III fyziologie MeSH
- kognice fyziologie MeSH
- kognitivní stárnutí * fyziologie MeSH
- lidé MeSH
- mozkový neurotrofický faktor krev MeSH
- neuropsychologické testy * statistika a číselné údaje MeSH
- paměť fyziologie MeSH
- prognóza MeSH
- průřezové studie MeSH
- regresní analýza MeSH
- senioři MeSH
- svalová atrofie etiologie MeSH
- svalová síla fyziologie MeSH
- Check Tag
- lidé MeSH
- senioři MeSH
- ženské pohlaví MeSH
- Publikační typ
- práce podpořená grantem MeSH
BACKGROUND: Maintaining healthy brain function during ageing is of great importance, especially for the self-sufficiency of older adults. The main aim of this study was to determine the effects of dance and martial arts on exerkines Brain Derived Neurotrophic Factor (BDNF) and irisin blood serum levels. METHODS: This randomized controlled trial examined the effects of dance and martial arts on serum Brain-Derived Neurotrophic Factor (BDNF) and irisin levels, as well as cognitive function, mood, and physical measures in older adults. Seventy-seven independently living older adults (mean age 70.3±3.8 years) were randomized into three groups: dance (DG), martial arts (MaG), and control (CG), followed over 12 weeks. Generalized linear models were used to assess the interventions' effects. RESULTS: There was a significant increase in BDNF levels in both the DG (1.8 ± 4.9, p < 0.05) and MaG (3.5 ± 6.3, p < 0.05), while CG experienced a decrease (-4.9 ± 8.2, p < 0.05). Between-group effects were significant for BDNF, with DG and MaG showing higher levels than CG (p < 0.05). No significant changes in irisin levels were found. Cognitive performance, particularly attention and mental flexibility (measured by the Trail Making Test A and B), significantly improved in the DG compared to CG (p < 0.05). Additionally, participants in DG showed improved mood based on the Geriatric Depression Scale (p < 0.05) compared to CG. Anthropometric T-scores were significantly associated with changes in irisin levels (p < 0.05) after intervention. CONCLUSION: The study found that dance and martial arts upregulated BDNF levels, with dance showing notable improvements in cognitive function and mood in older adults. Changes in anthropometric measures were linked to increased irisin levels. These findings suggest that both dance and martial arts may promote healthy brain function in aging populations. TRIAL REGISTRATION: NCT05363228.
- MeSH
- afekt MeSH
- bojové sporty * fyziologie MeSH
- fibronektiny * krev MeSH
- kognice * MeSH
- lidé MeSH
- mozkový neurotrofický faktor * krev MeSH
- senioři MeSH
- tanec * fyziologie MeSH
- tělesná výkonnost * fyziologie MeSH
- Check Tag
- lidé MeSH
- mužské pohlaví MeSH
- senioři MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- randomizované kontrolované studie MeSH
The search for the "Holy Grail" in clinical diagnostic microbiology-a reliable, accurate, low-cost, real-time, easy-to-use method-has brought up several methods with the potential to meet these criteria. One is Raman spectroscopy, an optical, nondestructive method based on the inelastic scattering of monochromatic light. The current study focuses on the possible use of Raman spectroscopy for identifying microbes causing severe, often life-threatening bloodstream infections. We included 305 microbial strains of 28 species acting as causative agents of bloodstream infections. Raman spectroscopy identified the strains from grown colonies, with 2.8% and 7% incorrectly identified strains using the support vector machine algorithm based on centered and uncentred principal-component analyses, respectively. We combined Raman spectroscopy with optical tweezers to speed up the process and captured and analyzed microbes directly from spiked human serum. The pilot study suggests that it is possible to capture individual microbial cells from human serum and characterize them by Raman spectroscopy with notable differences among different species. IMPORTANCE Bloodstream infections are among the most common causes of hospitalizations and are often life-threatening. To establish an effective therapy for a patient, the timely identification of the causative agent and characterization of its antimicrobial susceptibility and resistance profiles are essential. Therefore, our multidisciplinary team of microbiologists and physicists presents a method that reliably, rapidly, and inexpensively identifies pathogens causing bloodstream infections-Raman spectroscopy. We believe that it might become a valuable diagnostic tool in the future. Combined with optical trapping, it offers a new approach where the microorganisms are individually trapped in a noncontact way by optical tweezers and investigated by Raman spectroscopy directly in a liquid sample. Together with the automatic processing of measured Raman spectra and comparison with a database of microorganisms, it makes the whole identification process almost real time.
- MeSH
- algoritmy MeSH
- lidé MeSH
- optická pinzeta MeSH
- pilotní projekty MeSH
- Ramanova spektroskopie * metody MeSH
- sepse * MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
BACKGROUND: This pilot study aimed to investigate how fixed orthodontic appliances simultaneously applied on the upper and lower arches affect the oral environment in the medium term. METHODS: The oral status of 30 orthodontic patients was evaluated using the number of decay-missing-filled teeth (DMFT), plaque (PI), and gingival indices (GI) before bonding of fixed orthodontic appliances (T0) and during the therapy (T1). Besides, the gingival crevicular fluid (GCF) and a dental plaque were collected. Samples were analyzed for selected Candida sp. and for 10 selected oral bacteria using mass spectroscopy and multiplex polymerase chain reaction, respectively. RESULTS: In 60% of patients, deterioration of the oral status (demonstrated by the increase in PI) was recorded (p < 0.05). Moreover, the changes in PI correlated with those of GI (p < 0.001). At the T1 time point, the mean representation of Actinomyces sp. in the total prokaryotic DNA in GCF and dental plaque of individual patients increased compared to T0 (p < 0.05). The probability of finding any of the 7 selected periodontal bacteria combined with Candida sp. was 10 times higher in patients in whom PI deteriorated between T0 and T1 (p < 0.01). CONCLUSIONS: Changes in the oral microbial diversity and an increase in PI were observed in the medium term after bonding of orthodontic appliance. Our study highlights the importance of a complex approach in this type of research as the association between clinical characteristics and combined microbial parameters is higher than when evaluated separately.
Severe Early Childhood Caries (sECC) is a multifactorial disease associated with the occurrence of specific oral microorganisms and other environmental, behavioral, and genetic factors. This study aimed to construct a multivariable model including the occurrence of Candida spp. and selected behavioral factors (length of breastfeeding, serving sweet beverages and beginning of brushing child's teeth) to determine their relationships to the occurrence of sECC. In this case-control study 164 children with sECC and 147 children without dental caries were included. MALDI-TOF MS and multiplex qPCR were used to identify Candida spp. and selected bacteria in dental plaque samples, respectively. A questionnaire on oral hygiene, diet, and children's health was filled in by the parents. The constructed multivariable logistic regression model showed an independent influence of the microbial and behavioral factors in sECC etiopathogenesis. The occurrence of C. albicans and C. dubliniensis was associated with higher odds of sECC development (odds ratio, OR: 9.62 and 16.93, respectively), together with breastfeeding of 6 months or less (OR: 2.71), exposure to sweet beverages (OR: 3.77), and starting to brush child's teeth after the 12th month of age (OR: 4.10), all statistically significant (p < 0.01). Considering the high occurrence of C. albicans and C. dubliniensis in dental plaque in children with sECC, we propose them as "keystone pathogens" and risk factors for sECC. The models showed that presence of specific species of Candida in dental plaque may be a better descriptor of sECC than the mentioned behavioral factors.
- MeSH
- Candida albicans MeSH
- Candida MeSH
- dítě MeSH
- lidé MeSH
- náchylnost k zubnímu kazu MeSH
- předškolní dítě MeSH
- Streptococcus mutans MeSH
- studie případů a kontrol MeSH
- zubní kaz * MeSH
- zubní plak * MeSH
- Check Tag
- dítě MeSH
- lidé MeSH
- předškolní dítě MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Rapid and accurate identification of pathogens causing infections is one of the biggest challenges in medicine. Timely identification of causative agents and their antimicrobial resistance profile can significantly improve the management of infection, lower costs for healthcare, mitigate ever-growing antimicrobial resistance and in many cases, save lives. Raman spectroscopy was shown to be a useful-quick, non-invasive, and non-destructive -tool for identifying microbes from solid and liquid media. Modifications of Raman spectroscopy and/or pretreatment of samples allow single-cell analyses and identification of microbes from various samples. It was shown that those non-culture-based approaches could also detect antimicrobial resistance. Moreover, recent studies suggest that a combination of Raman spectroscopy with optical tweezers has the potential to identify microbes directly from human body fluids. This review aims to summarize recent advances in non-culture-based approaches of identification of microbes and their virulence factors, including antimicrobial resistance, using methods based on Raman spectroscopy in the context of possible use in the future point-of-care diagnostic process.
- MeSH
- analýza jednotlivých buněk MeSH
- antiinfekční látky * MeSH
- faktory virulence MeSH
- lidé MeSH
- Ramanova spektroskopie * metody MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- přehledy MeSH
Physical exercise may activate a number of important biochemical processes in the human body. The aim of this systematic review and meta-analysis was to identify the long-term effect of physical activity on irisin blood levels. We searched PubMed, Scopus, and Web of Science for articles addressing the long-term effect of physical exercise on irisin blood levels. Fifty-nine articles were included in the final qualitative and quantitative syntheses. A statistically significant within-group effect of exercise on irisin blood levels was in 33 studies; out of them, the irisin level increased 23× and decreased 10×. The significant positive between-groups effect was found 11×. Furthermore, the meta-analysis indicated that physical exercise had a significant positive effect on irisin blood levels (SMD = 0.39 (95% CI 0.27-0.52)). Nevertheless, considerably high heterogeneity was found in all the analyses. This systematic review and meta-analysis indicate that physical exercise might increase irisin blood levels; however, the results of individual studies were considerably inconsistent, which questions the methodological detection of irisin by ELISA kits.
- Publikační typ
- časopisecké články MeSH
- přehledy MeSH
- MeSH
- antibakteriální látky MeSH
- biofilmy * MeSH
- infekce močového ústrojí * MeSH
- lidé MeSH
- životní styl MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- úvodníky MeSH
Actinotignum schaalii is an emerging, opportunistic pathogen and its connection to non-infectious diseases and conditions, such as prostate or bladder cancer, or chronic inflammation has been proposed. Here, we analyzed 297 urine, ureteral and urinary catheter samples from 128 patients by Polymerase Chain Reaction followed by Denaturing Gradient Gel Electrophoresis and Sequencing (PCR-DGGE-S), and culture, and 29 of these samples also by 16S rRNA Illumina sequencing, to establish A. schaalii's prevalence in urinary tract-related samples, its relation to other bacteria, and its potential association with patients' conditions and samples' characteristics. A. schaalii-positive samples were significantly more diverse than A. schaalii negative and between-group diversity was higher than intra-group. Propionimicrobium lymphophilum, Fusobacterium nucleatum, Veillonella sp., Morganella sp., and Aerococcus sp. were significantly more often present in A. schaalii-positive samples; thus, we suggest these species are A. schaalii's concomitants, while Enterobacter and Staphylococcaceae were more often identified in A. schaalii-negative samples; therefore, we propose A. schaalii and these species are mutually exclusive. Additionally, a significantly higher A. schaalii prevalence in patients with ureter stricture associated hydronephrosis (p = 0.020) was noted. We suggest that A. schaalii could be an early polybacterial biofilm colonizer, together with concomitant species, known for pro-inflammatory features.
- Publikační typ
- časopisecké články MeSH
