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Abstract: Introduction: Low microbial biomass and high human DNA content in nasopharyngeal aspirate samples hinder comprehensive characterization of microbiota and resistome. We obtained samples from premature infants, a group with increased risk of developing respiratory disorders and infections, and consequently frequent exposure to antibiotics. Our aim was to devise an optimal protocol for handling nasopharyngeal aspirate samples from premature infants, focusing on host DNA depletion and microbiome and resistome characterization. Methods: Three depletion and three DNA extraction protocols were compared, using RT-PCR and whole metagenome sequencing to determine the efficiency of human DNA removal, taxonomic profiling and assignment of antibiotic resistance genes. Protocols were tested using mock communities, as well as pooled and individual patient samples. Results: The only extraction protocol to retrieve the expected DNA yield from mock community samples was based on a lytic method to improve Gram positive recovery (MasterPure™). Host DNA content in non-depleted aliquots from pooled patient samples was 99%. Only samples depleted with MolYsis™ showed satisfactory, but varied reduction in host DNA content, in both pooled and individual patient samples, allowing for microbiome and resistome characterisation (host DNA content from 15% to 98%). Other depletion protocols either retrieved too low total DNA yields, preventing further analysis, or failed to reduce host DNA content. By using Mol_MasterPure protocol on aliquots from pooled patient samples, we increased the number of bacterial reads by 7.6 to 1,725.8-fold compared to non-depleted reference samples. PCR results were indicative of achieved microbial enrichment. Individual patient samples processed with Mol_MasterPure protocol varied greatly in total DNA yield, host DNA content (from 40% to 98%), species and antibiotic resistance gene richness. Discussion: Despite high human DNA and low microbial biomass content in nasopharynx aspirates of preterm infants, we were able to reduce host DNA content to levels compatible with downstream shotgun metagenomic analysis, including bacterial species identification and coverage of antibiotic resistance genes. Whole metagenomic sequencing of microbes colonizing the nasopharynx may contribute to explaining the possible role of airway microbiota in respiratory conditions and reveal carriage of antibiotic resistance genes.
Keywords: antimicrobial resistance, host DNA depletion, low biomass, microbiome, premature infant, resistome, respiratory microbiome, whole metagenomic sequencing
Published in DiRROS: 25.02.2026; Views: 90; Downloads: 25
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Abstract: Background: MicroRNAs (miRNAs) are involved in the synthesis of proprotein convertase subtilisin–kexin type 9 (PCSK9), one of the regulators of low-density lipoprotein cholesterol (LDL-C) metabolism, and are directly involved in the atherosclerotic process. The aim of this study was to verify whether treatment with PCSK9 inhibitors (PCSK9i) and changes in the expression of miRNAs involved in PCSK9 metabolism are associated with arterial wall properties in stable post-myocardial infarction (MI) patients with insufficiently regulated LDL-C levels and significantly increased Lp(a) levels. Methods: Ninety-five patients after MI were enrolled and randomized to a placebo (N = 31) or PCSK9i group (N = 64). The treatment group received subcutaneous alirocumab 150 mg or evolocumab 140 mg, every 2 weeks. Blood for biochemical and epigenetic analysis was taken and ultrasound measurements of flow-mediated dilation of brachial artery (FMD), carotid intima–media thickness (c-IMT) and pulse wave velocity (PWV) were performed initially and after 6 months of treatment. The expression of the selected 5 miRNAs (miR-191-5p, miR-224-5p, miR-337-3p, miR-483-5p, and miR-552-3p) was quantified using quantitative polymerase chain reaction. Results: A decrease in c-IMT was associated with a decrease in the expression of miR-337-3p (ρ = 0.329; p = 0.010) and miR-483-5p (ρ = 0.324; p = 0.012). We did not detect any associations between miRNA changes and FMD or PWV. Conclusions: Our results suggest that changes in the selected miRNAs are associated with changes in the morphological properties of the arterial wall. We have shown that the decrease in miR-483-5p expression present a good indicator of the regression of morphological atherosclerotic change. The trial registration: The study is registered with CinicalTrials under the number NCT04613167, date of registration November 2nd, 2020. Approval for this study was obtained from the National Medical Ethics Committee of the Republic of Slovenia (reference number: KME 0120-357/2018/8).
Keywords: miRNA, endothelial function, PCSK9 inhibitors, myocardial infarction, low-density lipoprotein cholesterol
Published in DiRROS: 18.02.2026; Views: 195; Downloads: 68
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Abstract: Introduction: The purpose of this national research was to compare birth, maternal and newborn outcomes in the midwife led unit and the obstetric unit to ascertain whether a midwife led unit reduced medicalisation of childbirth. Methods: A prospective observational case-control study was carried out in Ljubljana Maternity Hospital in the period May - August 2013. The sample comprised 497 labouring women; 154 who attended the midwife led and 343 who attended in the obstetric unit, both matching the same inclusion criteria: low risk primiparous; singleton term pregnancies, normal foetal heart beat, cephalic presentation; spontaneous onset of labour. The primary outcome was the caesarean section rate. Chi-square test was used to compare medical interventions and birth outcomes. Results: Women in the midwife led unit had statistically significant higher spontaneous vaginal births (p < 0.001), less augmentation with oxytocin (p < 0.001), less use of analgesia (p < 0.001), less operative vaginal deliveries (p < 0.001) and less caesarean sections (p < 0.001), lower rates of episiotomy (p < 0.001) and more exclusively breastfed (p = 0.002). Discussion and conclusion: These significant findings showed that in the midwife led unit fewer medical interventions were used. For generalisation of the findings more similar studies in Slovenia are needed.
Keywords: low risk, labour, medical interventions, perinatal outcomes
Published in DiRROS: 28.01.2026; Views: 182; Downloads: 56
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Abstract: Calcium sulfoaluminate (CSA)-based cements represent a promising alternative to traditional Portland cement, offering benefits such as rapid strength development, low shrinkage, and reduced carbon footprint. Despite their commercial availability and proven performance, the adoption of CSA cements remains hindered by gaps in standardization and understanding. This RILEM Technical Committee (TC) CSA aims to review current knowledge, identify research needs, and address challenges related to clinkering, hydration mechanisms, durability, applications, nomenclature, standardization, and testing of CSA-based cements. The outcomes will further the understanding of manufacturing, technical performance, use, and specification of CSA-based cements, ultimately leading to broader acceptance in concrete construction.
Keywords: calcium sulfoaluminate cement, ye'elimite, belite, ettringite, low-CO2 cement
Published in DiRROS: 14.01.2026; Views: 163; Downloads: 97
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Abstract: Background and aims: Sex differences in cholesterol levels are well documented in adults and adolescents, but limited data exist for prepubertal children. This study aimed to evaluate innate sex differences in total cholesterol (TC) and low-density lipoprotein cholesterol (LDL-C) levels among prepubertal children, both in the general population and among those with familial hypercholesterolemia (FH). Methods: This cross-sectional study used data from Slovenia’s Universal FH Screening Program. Two populationbased random samples of children undergoing routine cholesterol screening at age 5 years were included from 2014 (N = 3412) and 2023 (N = 4182). In addition, a referred cohort from the Slovenian Hypercholesterolemia Registry (n = 1160, aged <10 years) who underwent genetic testing was analyzed. Results: In both the 2014 and 2023 cohorts, girls had significantly higher TC levels than boys (median difference: 0.10–0.11 mmol/L; p < 0.05). Among FH-negative children in the Registry, girls had on average 0.14 mmol/L higher TC and 0.13 mmol/L higher LDL-C than boys (both p < 0.05). No sex differences were observed in FHpositive children (p = 0.83 for TC; p = 0.82 for LDL-C). In the overall Registry cohort, after adjusting for FH status, girls had 0.11 mmol/L higher TC and 0.10 mmol/L higher LDL-C (both p < 0.05). Conclusion: Prepubertal girls have modestly higher TC and LDL-C than boys, a difference not observed in prepubertal FH-positive children, suggesting that the presence of a pathogenic FH variant may override innate physiological differences in lipid metabolism. These findings support universal early cholesterol screening and suggest that sex-specific reference values may improve early cardiovascular risk assessment in prepubertal FHnegative children.
Keywords: sex differences, prepubertal children, total cholesterol, low-density lipoprotein cholesterol, familial hypercholesterolemia
Published in DiRROS: 17.11.2025; Views: 466; Downloads: 181
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