Fakultní nemocnice Hradec Králové

Tuberculosis (TB) remains a significant global health challenge due to the rapid emergence of drug resistance. Despite substantial progress in anti-TB drug development, effective treatment options are limited. In this study, we report the synthesis and biological evaluation of pyrazinamide (PZA) derivatives with 5-alkyl and 5-alkanamido modifications, designed to enhance antimycobacterial activity by increasing lipophilicity and improving penetration of the lipid-rich mycobacterial cell wall. A positive correlation between the length of the 5-alkyl chain and antimycobacterial activity was observed, with maximal potency achieved with the heptyl substituent (4: 5-heptylpyrazine-2-carboxamide, MIC_M. tuberculosis H37Rv = 3.13 μg/mL). In series C with phenyl substitution on the C-2 carboxamide, different simple substituents were tolerated on the benzene ring (both electron-donating and electron-withdrawing, both lipophilic and hydrophilic), and the length of the alkyl chain was the main determinant of the antimycobacterial activity. Compound 23 (5-hexyl-N-(3-trifluoromethylphenyl)-pyrazine-2-carboxamide) exerted MIC = 3.13 μg/mL and selectivity index (SI, compared to HepG2 cells) >25. Notably, the tested compounds exhibited significant activity against multidrug-resistant (MDR) Mycobacterium tuberculosis strains while maintaining favorable selectivity profiles and low cytotoxicity. In contrast, 5-alkanamido derivatives (series B and D) were devoid of antimycobacterial activity. Mechanistic investigations revealed that unlike PZA, the 5-alkyl pyrazinamide derivatives are not hydrolyzed by mycobacterial pyrazinamidase (PncA), indicating a distinct mode of action. While molecular modeling initially suggested enoyl-ACP reductase (InhA) as a potential target of series C, subsequent experimental validation disproved this hypothesis; thus, the precise mechanism of action remains to be elucidated.

Preeclampsia is a major pregnancy complication that results in significant maternal and infant mortality and morbidity, yet difficulties remain in the diagnosis of preeclampsia based on clinical parameters alone. The objective was to assess the performance of a hand-held point-of-care (POC) immunoassay in a clinical environment for glycosylated fibronectin (GlyFn) for the prediction of preeclampsia within 4 weeks of sampling.

Multinational European prospective observational pilot study of predominantly high-risk patients in the second half of pregnancy to assess a point-of-care immunoassay for GlyFn in predicting preeclampsia within 4 weeks of sampling. GlyFn was measured using a second generation hand held POC immunoassay. Results were considered normal for GlyFn concentrations of < 350 µg/mL, positive for GlyFn concentrations of 351-600 µg/mL, and high-positive for GlyFn concentrations > 600 µg/mL.

Preeclampsia developed in 16 (19%) of 84 subjects and was associated with a shorter gestational age at delivery 35.3 weeks vs. 37.3 weeks for non-preeclamptics, n = 82; p = 0.001), a higher risk of fetal growth restriction (FGR; 31.2% vs. 10.3% for non-preeclamptics, p = 0.046), and an increased risk of preterm birth < 37 weeks gestation (83.3% vs. 33.3% for non-preeclamptics, (n = 78; p = 0.003). GlyFn positive or high positive was seen in 13/16 (81%) and in 35/68 (51.5%), yielding a sensitivity of 81%, a specificity of 49%, a positive predictive value of 27%, and a negative predictive value of 92%. GlyFn positive or high positive was also associated with preterm birth < 37 weeks in singleton pregnancy non-preeclamptic patients. Preterm birth occurred in 4.8% of those with normal GlyFn, in 26.7% with positive GlyFn, and in 50% of those with high GlyFn in singleton gestations without preeclampsia (p = 0.008).

The ability to use this test in a POC format provides a method for practitioners to quickly determine risk for preeclampsia in their pregnant patients and offers an affordable alternative, as a single analyte to other diagnostic or screening tests that require laboratory-based testing or ultrasound equipment. Independent of preeclampsia, an elevated GlyFn was also correlated with preterm delivery and requires further study.

As part of a broader survey of lignicolous saprobic fungi, we investigated fungal taxa from the class Sordariomycetes displaying holoblastic-denticulate conidiogenesis, a distinct developmental process and phylogenetically informative trait. Although these fungi appear morphologically similar in culture, they represent distinct evolutionary lineages. This taxonomic study integrates comparative morphological analyses, phylogenetic to introduce novel taxa in the Pleurotheciales and Rhamphoriales . A new genus and species Echinodenticula allantospora and three new species, Phaeoisaria parallela , Rhamphoriopsis cuprea and Rh. denticulata , are described. A rarely encountered species Rhamphoria separata is reported, along with its previously undocumented asexual morph. Furthermore, we successfully demonstrate the utility of two protein-coding genes, rpb2 and tef1 , as complementary barcodes for distinguishing closely related Phaeoisaria Rhamphoriales and a prevalent trait in the Pleurotheciales . An unknown ascomycete that produced only sterile mycelium in culture is described here as Melanocrypta curvata and placed at an incertae sedis position within the Sordariomycetes . Additionally, we present short-read whole-genome sequencing data for the ex-type strains of the newly described species, providing a valuable genomic resource for future taxonomic, phylogenetic, and functional studies. Environmental DNA data from the GlobalFungi database bring new perspective into the biogeographical patterns of Phaeoisaria , Rhamphoria , and Rhamphoriopsis . The distribution of E. allantospora and M. curvata remains poorly understood, as no records for these species were found in GlobalFungi. This study provides new insights into the molecular systematics, taxonomy, and biogeography of the Rhamphoriales and Pleurotheciales , and highlights the role of environmental DNA metabarcoding in uncovering fungal diversity and distribution patterns.