The aim of this prospective study was to assess the duration of culture-viable SARS-CoV-2 and to monitor the emergence of mutations in a cohort of 23 kidney transplant recipients (KTRs) from June 2022 to June 2023. Combined nares/oropharyngeal swabs were collected weekly starting as soon as possible after symptom onset. The time from symptom onset to a negative culture was 11 days (interquartile range, 8-14), while the time to negative reverse transcriptase quantitative polymerase chain reaction was 18 days (interquartile range, 15-30). Beyond the first swab, 21.7% had a positive culture, and 8.7% replicated viable virus for longer than 30 days. T cell depletion (rate ratio, 2.5; 95% confidence interval [95% CI], 1.9-3.3; P < .001) and time from transplantation (rate ratio, 0.93; 95% CI, 0.90-0.97; P = .006) were associated with the time of viable virus shedding. A cycle threshold value of 24.2 demonstrated a 91.3% negative predictive value of viability (95% credible interval [95% CrI], 76-100). The odds of viability decreased by 69% per week of infection (odds ratio, 0.31; 95% CrI, 0.12-0.76). Overall, ribonucleic acid sequencing did not show accelerated molecular evolution though mutation rate could be increased in molnupiravir-treated KTRs. In conclusion, viable SARS-CoV-2 is eliminated rapidly, the risk of virus evolution is low, and prolonged self-isolation is generally unnecessary for most KTRs.

• MeSH
• COVID-19 * virologie   epidemiologie   MeSH
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• molekulární evoluce * MeSH
• mutace MeSH
• příjemce transplantátu * MeSH
• prospektivní studie MeSH
• SARS-CoV-2 * genetika   izolace a purifikace   fyziologie   MeSH
• senioři MeSH
• transplantace ledvin * MeSH
• vylučování virů * MeSH
• Check Tag
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• mužské pohlaví MeSH
• senioři MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• pozorovací studie MeSH

The largest obstacle in the promotion of biopesticides is the existence of counterfeit products available in the market. Identification and quantification of antagonistic organisms in biopesticide products are the key to the reduction of spurious microbial pesticides. In this study, we have developed a simple, sensitive, isothermal-based colourimetric assay for specific detection of Bacillus subtilis from the biopesticide formulations and soil samples. A region specific to B. subtilis which codes for shikimate dehydrogenase was identified through in silico analysis. We employed conventional PCR, loop-mediated isothermal amplification (LAMP), recombinase polymerase amplification (RPA), and qPCR for specific detection of B. subtilis in soil samples and biopesticide formulations. Specificity tests showed that the PCR primers amplified an amplicon of 521 bp in four strains of B. subtilis only, and no amplification was found in negative control samples. Similarly, the LAMP assay showed sky blue colour in all four strains of B. subtilis and violet colour in negative control samples. Whereas in the RPA assay, upon the addition of SYBR Green dye, a bright green colour was seen in B. subtilis strains, while a brick-red colour was observed in negative control samples by visualizing under a UV transilluminator. The qPCR assay showed specific amplifications with a Ct value of 12 for B. subtilis strains and no amplification in negative control samples. In the sensitivity test, PCR could amplify DNA of B. subtilis up to 500 pg/μL. DNA concentration as low as 10 pg/μL was enough to show the colour change in the LAMP as well as the RPA assays, whereas the qPCR assay showed sensitivity till 100 pg/μL. All four diagnostic assays developed in the study have been validated in soil samples and B. subtilis-based biopesticides. Compared to conventional PCR, the qPCR assay has the advantage of quantification and visualizing the result in real-time, whereas LAMP and RPA assays have the benefits of being colourimetric and less time-consuming. The other advantages are that the results can be visualized with the naked eye, and these assays do not require a costly thermal cycler and gel documentation system. Hence, LAMP and RPA assays are highly suitable for developing point-of-need diagnostic kits and, in turn, help regulators assess the quality of biopesticides in the market.

• MeSH
• alkoholoxidoreduktasy * genetika   MeSH
• Bacillus subtilis * genetika   izolace a purifikace   enzymologie   MeSH
• bakteriální proteiny genetika   MeSH
• diagnostické techniky molekulární * metody   MeSH
• kolorimetrie * metody   MeSH
• kvantitativní polymerázová řetězová reakce MeSH
• půdní mikrobiologie MeSH
• senzitivita a specificita MeSH
• techniky amplifikace nukleových kyselin * metody   MeSH
• Publikační typ
• časopisecké články MeSH

INTRODUCTION: Plasma phosphorylated tau (p-tau)181, glial fibrillary acidic protein (GFAP), neurofilament light chain (NfL), and amyloid beta ratio (Aβ42/40) may have diagnostic and prognostic value in Alzheimer's disease (AD). Here we assess which markers can best identify AD from controls and other non-AD dementias in a large international multi-center study. METHODS: Plasma samples (n = 1298) were collected from six international centers. Aβ40, Aβ42, GFAP, NfL, and p-tau181 were measured using single molecule array. In each group, AD diagnosis/co-pathology was defined according to cerebrospinal fluid biomarkers or amyloid positron emission tomography. Validations were performed in three separate cohorts via single and dual cut-off models. RESULTS: p-tau181 showed the best area under the curve value to separate AD from frontotemporal dementia, controls, and Aβ- dementia with Lewy bodies. However, this discriminative power could not be reproduced by applying pre-defined cut-offs. DISCUSSION: p-tau181 was the best single plasma marker for detecting AD at any stage. Specific cut-offs are needed to maximize diagnostic performances. HIGHLIGHTS: Phosphorylated tau (p-tau)181 provided a clear differentiation between controls and Alzheimer's disease (AD) participants, with evidence of increased levels in the preclinical stage of AD. Plasma biomarkers demonstrated that when amyloid co-pathology is removed from dementia with Lewy bodies (DLB), only glial fibrillary acidic protein and neurofilament light chain remain to predict DLB. Given the low prevalence of amyloid co-pathology in frontotemporal dementia (FTD), p-tau181 and its ratio with amyloid beta 42 are strong biomarkers to differentiate FTD from AD.

• MeSH
• Alzheimerova nemoc * krev   diagnóza   MeSH
• amyloidní beta-protein * krev   MeSH
• biologické markery krev   MeSH
• demence * krev   diagnóza   MeSH
• fosforylace MeSH
• gliový fibrilární kyselý protein * krev   MeSH
• lidé středního věku MeSH
• lidé MeSH
• neurofilamentové proteiny * krev   MeSH
• peptidové fragmenty * krev   MeSH
• proteiny tau * krev   MeSH
• senioři MeSH
• Check Tag
• lidé středního věku MeSH
• lidé MeSH
• mužské pohlaví MeSH
• senioři MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• multicentrická studie MeSH

INTRODUCTION: Chitinase-3-like protein 1 (CHI3L1) is a glycoprotein implicated in various neurological conditions. It is associated with neuroinflammation and tissue remodeling. The study aimed to validate the reference interval (RI) of serum (S) CHI3L1 in a control group, to correlate S CHI3L1 values with other biomarkers of neurodegenerative damage, and to estimate the diagnostic accuracy of S CHI3L1. METHODS: Samples from 108 healthy volunteers were used to estimate the S CHI3L1 RI. For the comparison, we used cerebrospinal fluid (CSF) and serum (S) samples from 121 patients with cognitive disorders, and cognitive deterioration was assessed using the Mini-Mental State Examination (MMSE). ELISA assays were used to determine the S CHI3L1, CSF, and S neurofilament light chain (NfL) levels; CSF and plasma β-amyloid peptide42; CSF and plasma β-amyloid peptide40; CSF total tau protein; CSF phosphorylated tau protein; and CSF alpha-synuclein. RESULTS: The estimated RI of S CHI3L1 was 14.44 to 63.11 μg/L. The cut-off value of S CHI3L1 was 34.37 μg/L. ROC analysis showed that S CHI3L1 has 81.4% sensitivity and 76.9% specificity. We found a moderate Spearman's rank correlation coefficient between the S CHI3L1 and age (rS = 0.486; p < 0.001) and between S CHI3L1 and S NfL (rS = 0.489; p < 0.001) in all groups. The Kruskal-Wallis test showed a significant overall difference in S CHI3L1 among diagnostic groups (p = 0.013). S CHI3L1 and CSF NfL had statistically significant effects on MMSE values (multiple R2 was 0.431). CONCLUSIONS: Our results suggest that S CHI3L1 reflects the severity of cognitive deficits assessed by MMSE. It can be used as a supportive biomarker in neurodegenerative diseases.

• MeSH
• alfa-synuklein mozkomíšní mok   krev   MeSH
• amyloidní beta-protein krev   mozkomíšní mok   MeSH
• biologické markery krev   mozkomíšní mok   MeSH
• dospělí MeSH
• kognitivní dysfunkce * krev   mozkomíšní mok   diagnóza   MeSH
• lidé středního věku MeSH
• lidé MeSH
• neurofilamentové proteiny mozkomíšní mok   krev   MeSH
• pohybové poruchy * krev   mozkomíšní mok   diagnóza   MeSH
• protein CHI3L1 * krev   mozkomíšní mok   MeSH
• proteiny tau mozkomíšní mok   krev   MeSH
• senioři nad 80 let MeSH
• senioři 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
• srovnávací studie MeSH

Preoperative differentiation between responders and non-responders to ventriculoperitoneal (VP) shunting in the treatment of normal pressure hydrocephalus (NPH) remains a significant challenge. Identifying biomarkers in presurgical assessment represents a promising approach to reducing the need for invasive cerebrospinal fluid CSF testing. In this prospective observational study, thirty adult patients were classified into Group A (responders to VP shunting) and Group B (non-responders) based on their responsiveness to invasive CSF testing. The overall clinical condition and Idiopathic NPH (iNPH) scale were assessed at baseline. Additionally, biomarker levels were compared between the two groups. Elevated levels of Neurofilament Light Chain (NfL) and Neurofilament Heavy Chain (NfH) in CSF and a reduced level of beta-amyloid Aβ42 were observed. No significant differences in biomarker levels were found between groups. Individual biomarkers demonstrated only poor predictive value (AUC = 0.37-0.53). Clinical factors were stronger predictors (AUC = 0.642-0.669), with no improvement when combined with all examined biomarkers (AUC = 0.428-0.431). No single biomarker reliably predicted confirmed postoperative shunt responsiveness among patients who underwent VP shunt placement and demonstrated clinical improvement. Clinical factors were stronger predictors, suggesting that patient history and clinical assessment (e.g., the iNPH scale) provide more reliable diagnostic information. Notably, combining biomarkers with clinical factors did not improve predictive accuracy.

• MeSH
• amyloidní beta-protein mozkomíšní mok   MeSH
• biologické markery * mozkomíšní mok   MeSH
• lidé středního věku MeSH
• lidé MeSH
• neurofilamentové proteiny mozkomíšní mok   MeSH
• normotenzní hydrocefalus * chirurgie   mozkomíšní mok   diagnóza   MeSH
• peptidové fragmenty mozkomíšní mok   MeSH
• prediktivní hodnota testů MeSH
• prospektivní studie MeSH
• senioři nad 80 let MeSH
• senioři MeSH
• ventrikuloperitoneální zkrat * metody   MeSH
• výsledek terapie MeSH
• Check Tag
• lidé středního věku MeSH
• lidé MeSH
• mužské pohlaví MeSH
• senioři nad 80 let MeSH
• senioři MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• pozorovací studie MeSH

Chronic bronchitis is increasingly reported as a healthcare challenge in clinical settings partially due to the disease's bad prognosis and unresponsiveness to therapy, including the ineffectiveness of glucocorticoids. The ineffectiveness could have a link with genetic polymorphism of receptor genes resulting in inappropriate glucocorticoid pharmacodynamics. We sought to identify the role of gene polymorphism in the response of patients with chronic bronchitis to prednisolone therapy. To do so, a total of 60 newly diagnosed chronic bronchitis patients enrolled in the present study. Prednisolone at a dose of 30mg/day for two weeks was given and respiratory parameters [forced expiratory volume in 1 second (FEV1), forced vital capacity (FVC), and FEV1/FVC were measured before and after therapy. Blood samples were withdrawn for genetic profiling of genes involved in glucocorticoids pharmacodynamics, including BCII (rs41423247), N363S (rs56149945), and ER22/23EK (rs6189/rs6190) measured for their homozygous versus heterozygous gene splice variants.Results: Gene splice variants for BCII (rs41423247), N363S (rs56149945), and ER22/23EK (rs6189/rs6190) homozygous (73.3%, 98.7%, and 95%) represented a higher percentage than heterozygous (26.7%, 1.7%, and 5%). The respiratory parameters FEV1, FVC, and FEV1/FVC have shown significantly (p<0.05) better values at baseline in homozygous versus heterozygous, correspondingly, the responsiveness to therapy has shown significantly (p<0.05) better values in homozygous versus heterozygous.Conclusion: The study has provided a good template for genetic behaviour toward individualised medicine in our locality providing that these genes could be a cornerstone for discovering issues related to the pharmacodynamics profiling of drugs in clinical settings.

• MeSH
• chronická bronchitida * diagnóza   genetika   MeSH
• glukokortikoidy farmakologie   MeSH
• lidé MeSH
• polymerázová řetězová reakce metody   MeSH
• polymorfismus genetický genetika   MeSH
• prednisolon farmakologie   terapeutické užití   MeSH
• protein - isoformy genetika   MeSH
• receptory glukokortikoidů * genetika   účinky léků   MeSH
• respirační funkční testy metody   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• klinická studie MeSH
• práce podpořená grantem MeSH

Chem. Listy 2022; 116: 101−109. doi: 10.54779/ chl20220101 vladimir.pliska@biol.ethz.ch, parizek@porodnice.cz Došlo 12. 10. 2021, přijato 1. 11. 2021.

Abstract: Licence agreements between the Institute of Organic Chemistry and Biochemistry, Czechoslovak Academy of Sciences, Prague (IOCB), and the pharmaceutical company Ferring AB Malmö enabled the Swedish company to produce and commercialize worldwide a number of neurohypophyseal peptides designed at the IOCB. Several of them found therapeutic applications. dDAVP: 1-deamino-8-D-arginine-vasopressin was designed in one of the IOCB peptide laboratories (M. Zaoral and F. Šorm) in 1967. It displayed an extremely high antidiuretic activity (various tests indicate a 2- to 50-fold increase, as compared to arginine vasopressin) and a very low pressor activity. The peptide (covered by the U.S. Patent No. 3,497,491, 1970) has been used as a preferred drug in the substitution therapy of the central form of diabetes insipidus (Minirin®, today as Desmopressin INN). Besides, as later discovered (Mannucci et al. 1977), dDAVP increases the plasma concentration of the blood clottiing factor VIII. This fact extended its clinical use as haemostatics in cases of milder forms of haemophilia A, von Willebrand-Jürgens syndrome and some thrombocyte dysfunctions. Despite of the clinical success of dDAVP, a closer look reveals certain inadequacies in the presently available pharmacological data: several reports declare activity values and the prolongation effect (index of persistence) in very broad ranges. Triglycyl-8- -lysine-vasopressin (Terlipressin), a peptide with lysine vasopressin chain extended at the N-terminal by a triglycine residue, acts mainly as a prodrug (releasing lysine vasopressin after aminopeptidase splitting at the Na group). The analogue belongs to the so-called „synthetic hormonogens“; individual peptides carrying various acylating groups were synthesized in the midsixties at the IOCB and legally protected by U.S. Patent No. 3,558,590 (1968). It was a part of the licence agreements mentioned above. The activities of triglycyl-8-lysine-vasopressin (both antidiuretic and vasopressor) are about 100 times lower than those of lysine vasopressin, but its persistence is 5 times longer. As such, it is occasionally used in emergency medicine in cases of esophageal (and other gastroenteral) bleeding, traumatic or septic shock, in cirrhotic patients and patients with portal hypertension. Its use as an early abortion drug was discussed but not pursued. Carbetocin (deaminocarba1-2-O-methyltyrosin-oxytocin) was synthetized in the laboratory of Karel Jošt at the IOCB before 1971; its synthesis was covered by a Czechoslovak patent (CS-149,028 B1) in June 1973 (at that time, Czechoslovak patent law did not provide for the patentability of substances as such) and first published in a biophysical communication by Frič et al., 1974). As a part of the licence agreement, it was included in the production program of Ferring AB, but marketed later also by several other pharmaceutical companies due to an incomplete patent protection. The peptide is a moderately active uterotonic partial agonist and as such has been utilized in veterinary obstetrics for delivery induction in cows and (multiparous) pigs: its milder and better--controlled uterotonic action was found preferential as compared to oxytocin so far used for these purposes. In the last two decades, carbetocin has been commonly used also in the human obstetrics, especially to prevent the peri- and post-partum haemorrhage, from the maternal side the most frequent and most severe delivery complication. It became a life-saving drug in emergency obstetrics.

• Klíčová slova
• carbetocin,
• MeSH
• desmopresin aplikace a dávkování   dějiny   farmakologie   terapeutické užití   MeSH
• lidé MeSH
• neurohypofyzární hormony * dějiny   farmakologie   klasifikace   MeSH
• oxytocin antagonisté a inhibitory   dějiny   farmakologie   MeSH
• poporodní krvácení farmakoterapie   MeSH
• terlipresin aplikace a dávkování   dějiny   farmakologie   terapeutické užití   MeSH
• uterotonika aplikace a dávkování   dějiny   farmakologie   klasifikace   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• přehledy MeSH
• Geografické názvy
• Česká republika MeSH
• Švédsko MeSH

BACKGROUND: Diabetes mellitus (DM) is a chronic disease with prevalence increasing worldwide. The aim of this study was to investigate satisfaction with the current method of insulin delivery (INS) amongst patient with type 1 diabetes mellitus (T1DM) using multiple daily injection (MDI) or continuous subcutaneous insulin infusion (CSII). Furthermore, a sub-aim was to test the effect of selected variables on patient satisfaction with MDI or CSII using regression analysis. METHODS: A cross-sectional study carried out in the territory of Moravia in the Czech Republic. A quantitative approach using the Insulin Delivery System Rating Questionnaire (IDSRQ) among 197 respondents with T1DM with INS delivery with MDI or CSII for at least 1 year. Statistical methods used were descriptive statistics, Student's t-tests and regression analysis. RESULTS: Highly significant differences were found between CSII and MDI patients in satisfaction with the current method of INS delivery (p < 0.001), in how the current method of delivery helps patients maintain stable blood glucose values, prevent high blood glucose (p < 0.001), and in overall satisfaction with the current method of INS delivery (p < 0.001). The average overall satisfaction score was 56.19 points for MDI and 62.08 points for CSII. Regression analysis revealed predictors of overall satisfaction on the mean score on how the current method of INS delivery helps MDI patients (p < 0.01). The effect of other selected variables was not confirmed. CONCLUSION: The results of the study showed higher overall satisfaction with the method of INS delivery in CSII patients. The current method of INS delivery does not interfere with daily life and activities in most patients.

• MeSH
• diabetes mellitus 1. typu * farmakoterapie   krev   psychologie   MeSH
• dospělí MeSH
• hypoglykemika * aplikace a dávkování   terapeutické užití   MeSH
• injekce subkutánní MeSH
• inzulin * aplikace a dávkování   terapeutické užití   MeSH
• inzulinové infuzní systémy * MeSH
• krevní glukóza MeSH
• lidé středního věku MeSH
• lidé MeSH
• mladý dospělý MeSH
• průřezové studie MeSH
• průzkumy a dotazníky MeSH
• spokojenost pacientů * MeSH
• subkutánní infuze MeSH
• Check Tag
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• mladý dospělý MeSH
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• Geografické názvy
• Česká republika MeSH

OBJECTIVES: The aim of the study was to evaluate the efficiency of molecular diagnostics of tick-borne encephalitis (TBE) and to correlate viral RNA (vRNA) detection with the clinical and laboratory data. METHODS: Clinical samples from 1125 patients from South Bohemia, Czech Republic, a highly endemic TBE region, were screened for TBE virus (TBEV) RNA by RT-qPCR. Samples included blood, serum, cerebrospinal fluid (CSF), and urine. RESULTS: TBEV RNA was detected in 14 patients with clinically proven TBE. TBEV RNA was most frequently detected in sera during early infection (11/37 patients tested, 29.7%) but decreased with rising IgG antibody response (3/228, 1.3%). Detection in CSF and urine was infrequent (1/30, 3.3% and 1/52, 1.9%, respectively). Additionally, five patients initially not diagnosed with TBE were retrospectively found to have TBEV RNA in serum, indicating possible underdiagnosis, particularly in mild or atypical presentations. The study also highlighted the diagnostic challenge of an immunocompromised patient whose delayed antibody response hindered timely diagnosis. In such cases, RT-qPCR could significantly shorten the diagnostic timeline. CONCLUSIONS: These findings underscore the value of early RNA detection in improving the diagnosis of TBE and may in the future facilitate the early administration of potential treatment, thereby improving patient outcomes.

• MeSH
• diagnostické techniky molekulární metody   MeSH
• dítě MeSH
• dospělí MeSH
• imunoglobulin G krev   MeSH
• klíšťová encefalitida * diagnóza   virologie   MeSH
• kvantitativní polymerázová řetězová reakce MeSH
• lidé středního věku MeSH
• lidé MeSH
• mladiství MeSH
• mladý dospělý MeSH
• předškolní dítě MeSH
• protilátky virové krev   MeSH
• RNA virová * krev   mozkomíšní mok   izolace a purifikace   MeSH
• senioři nad 80 let MeSH
• senioři MeSH
• viry klíšťové encefalitidy * izolace a purifikace   genetika   MeSH
• Check Tag
• dítě MeSH
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• mladiství MeSH
• mladý dospělý MeSH
• mužské pohlaví MeSH
• předškolní dítě MeSH
• senioři nad 80 let MeSH
• senioři MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• Geografické názvy
• Česká republika MeSH

African trypanosomes are medically important parasites that cause sleeping sickness in humans and nagana in animals. In addition to their pathogenic role, they have emerged as valuable model organisms for studying fundamental biological processes. Protein tagging is a powerful tool for investigating protein localization and function. In a previous study, we developed two plasmids for rapid and reproducible polymerase chain reaction-based protein tagging in trypanosomes, which enabled the subcellular mapping of 89% of the trypanosome proteome. However, the limited selection of fluorescent protein tags and selectable markers restricted the flexibility of this approach. Here, we present an extended set of >100 plasmids that incorporate universal primer annealing sequences, enabling protein tagging with a range of fluorescent, biochemical and epitope tags, using five different selection markers. We evaluated the suitability of various fluorescent proteins for live and fixed cell imaging, fluorescent movies, and we demonstrate the use of tagging plasmids encoding tandem epitope tags to support expansion microscopy approaches. We show that this series of plasmids is functional in other trypanosomatid parasites, significantly increasing its value. Finally, we developed a new plasmid for tagging glycosylphosphatidylinositol-anchored proteins. We anticipate that this will be an important toolset for investigating trypanosomatid protein localization and function.

• MeSH
• lidé MeSH
• plazmidy genetika   MeSH
• protozoální proteiny * metabolismus   genetika   MeSH
• transport proteinů MeSH
• Trypanosoma brucei brucei metabolismus   genetika   MeSH
• Trypanosomatina * metabolismus   genetika   MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH