BACKGROUND: Human milk harbors diverse bacterial communities that contribute to infant health. Although pumping and storing milk is a common practice, the viable bacterial composition of pumped milk and the impact of storage practice on these bacteria remains under-explored. This metagenomic observational study aimed to characterize viable bacterial communities in freshly pumped human milk and its changes under different storage conditions. METHODS: In 2023, twelve lactating mothers from the CELSPAC: TNG cohort (Czech Republic) provided freshly pumped milk samples. These samples were stored under various conditions (refrigeration for 24 h, 48 h, or freezing for six weeks) and treated with propidium monoazide (PMA) to selectively identify viable cells. The DNA extracted from individual samples was subsequently analyzed using 16S rRNA amplicon sequencing on the Illumina platform. RESULTS: The genera Streptococcus, Staphylococcus, Diaphorobacter, Cutibacterium, and Corynebacterium were the most common viable bacteria in fresh human milk. The median sequencing depth and Shannon index of fresh human milk samples treated with PMA (+ PMA) were significantly lower than in untreated (-PMA) samples (p < 0.05 for all), which was true also for each time point. Also, significant changes in these parameters were observed between fresh human milk samples and their paired frozen samples (p < 0.05), while no differences were found between fresh human milk samples and those refrigerated for up to 48 h (p > 0.05). Of specific genera, only + PMA frozen human milk samples showed a significant decrease in the central log-ratio transformed relative abundances of the genera Diaphorobacter and Cutibacterium (p < 0.05) in comparison to + PMA fresh human milk samples. CONCLUSIONS: The study demonstrated that the bacterial profiles significantly differed between human milk samples treated with PMA, which represent only viable bacteria, and those untreated. While storage at 4 °C for up to 48 h did not significantly alter the overall diversity and composition of viable bacteria in human milk, freezing notably affected both the viability and relative abundances of some bacterial genera.

• MeSH
• azidy MeSH
• Bacteria * izolace a purifikace   genetika   klasifikace   MeSH
• chlazení MeSH
• dospělí MeSH
• lidé MeSH
• mateřské mléko * mikrobiologie   MeSH
• mikrobiota * MeSH
• propidium analogy a deriváty   MeSH
• RNA ribozomální 16S MeSH
• skladování potravin * metody   MeSH
• zmrazování MeSH
• Check Tag
• dospělí MeSH
• lidé MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• pozorovací studie MeSH

AIM: This study aimed to evaluate bacteriome profiles (diversity, composition and relative abundances of bacterial genera) of the fluids from apical periodontitis (AP)-related radicular cysts (RCs). METHODOLOGY: This observational study included 29 patients with AP and RC with complete sample triplets (supragingival plaque, cryopulverized tooth and cystic fluid). The bacteriome profiles of each matrix as well as of negative controls (NCs) were examined using 16S rRNA amplicon sequencing. RESULTS: Bacteriome profiles of cystic fluids differed from NCs in 79% of cases. The number of distinct amplicon sequence variants and Shannon index detected in cystic fluids and cryopulverized teeth were significantly lower than in paired supragingival plaque samples. Gram-negative genera and anaerobic genera were more abundant in cystic fluids than in paired cryopulverized teeth or their supragingival plaques. The relative abundances of the genera Prevotella_7/Prevotella, Fusobacterium and Porphyromonas were higher in cystic fluids than in paired cryopulverized teeth and NCs; their relative abundances dominated (>20%) in individual cystic fluids. Also, DNA from the genus Fretibacterium was significantly more commonly found in cryopulverized teeth and cystic fluids than in supragingival plaque samples. The relative abundances of this gram-negative bacterial genera in cryopulverized teeth differed from NCs; the difference from cystic fluids was borderline insignificant. CONCLUSIONS: Although the alpha-diversity in the cystic fluids is much lower compared to supragingival plaques, most cystic fluids are not sterile. DNA from specific anaerobic gram-negative bacterial genera dominated the fluids from AP-related RCs.

• MeSH
• Bacteria * izolace a purifikace   genetika   klasifikace   MeSH
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• periapikální periodontitida * mikrobiologie   komplikace   MeSH
• radikulární cysta * mikrobiologie   MeSH
• RNA ribozomální 16S genetika   MeSH
• zubní plak mikrobiologie   MeSH
• Check Tag
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• pozorovací studie MeSH

The antibiotic resistance genes (ARGs) limit the susceptibility of bacteria to antimicrobials, representing a problem of high importance. Current research on the presence of ARGs in microorganisms focuses mainly on humans, livestock, hospitals, or wastewater. However, the spectrum of ARGs in the dust resistome in workplaces and households has gone relatively unexplored. This pilot study aimed to analyze resistome in indoor dust samples from participants' workplaces (a pediatric hospital, a maternity hospital, and a research center) and households and compare two different approaches to the ARGs analysis; high-throughput quantitative PCR (HT-qPCR) and whole metagenome shotgun sequencing (WMGS). In total, 143 ARGs were detected using HT-qPCR, with ARGs associated with the macrolides, lincosamides, and streptogramin B (MLSB) phenotype being the most abundant, followed by MDR (multi-drug resistance) genes, and genes conferring resistance to aminoglycosides. A higher overall relative quantity of ARGs was observed in indoor dust samples from workplaces than from households, with the pediatric hospital being associated with the highest relative quantity of ARGs. WMGS analysis revealed 36 ARGs, of which five were detected by both HT-qPCR and WMGS techniques. Accordingly, the efficacy of the WMGS approach to detect ARGs was lower than that of HT-qPCR. In summary, our pilot data revealed that indoor dust in buildings where people spend most of their time (workplaces, households) can be a significant source of antimicrobial-resistant microorganisms, which may potentially pose a health risk to both humans and animals.

• MeSH
• antibakteriální látky farmakologie   MeSH
• Bacteria genetika   izolace a purifikace   účinky léků   klasifikace   MeSH
• bakteriální geny genetika   MeSH
• bakteriální léková rezistence genetika   MeSH
• charakteristiky rodiny MeSH
• kvantitativní polymerázová řetězová reakce MeSH
• lidé MeSH
• metagenom MeSH
• mikrobiologie vzduchu MeSH
• pilotní projekty MeSH
• prach * analýza   MeSH
• pracoviště * MeSH
• vysoce účinné nukleotidové sekvenování MeSH
• znečištění vzduchu ve vnitřním prostředí MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH

BACKGROUND: Mouth breathing (MB), a risk factor of oral dysbiosis and halitosis, is linked with craniofacial anomalies and pediatric obstructive sleep apnea. Here, we aimed to analyze tongue microbiota in children from the perspective of their breathing pattern before/during orthodontic treatment. METHODS: This prospective case-control study included 30 children with orthodontic anomalies, 15 with MB and 15 with nasal breathing (NB), matched by age, sex, and body mass index. All underwent orthodontic examination and sleep apnea monitoring. Tongue swabs were collected before starting (timepoint M0) and approx. six months into the orthodontic therapy (timepoint M6). Oral candidas and bacteriome were analyzed using mass spectrometry technique and 16S rRNA sequencing, respectively. RESULTS: MB was associated with higher apnea-hypopnea index. At M0, oral candidas were equally present in both groups. At M6, Candida sp. were found in six children with MB but in none with NB. No significant differences in bacterial diversity were observed between groups and timepoints. However, presence/relative abundance of genus Solobacterium was higher in children with MB than NB at M0. CONCLUSIONS: Significant links between MB and the presence of genus Solobacterium (M0) as well as Candida sp. (M6) were found in children with orthodontic anomalies, highlighting the risk of halitosis in them.

• MeSH
• dítě MeSH
• dýchání ústy * MeSH
• halitóza mikrobiologie   MeSH
• jazyk * mikrobiologie   MeSH
• lidé MeSH
• mikrobiota * MeSH
• prospektivní studie MeSH
• RNA ribozomální 16S analýza   MeSH
• studie případů a kontrol MeSH
• Check Tag
• dítě MeSH
• lidé MeSH
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH

Intrapartum antibiotic prophylaxis (IAP) is commonly used during C-section delivery and in Group B Streptococcus-positive women before vaginal delivery. Here, we primarily aimed to investigate the effect of IAP on the neonatal oral and fecal bacteriomes in the first week of life. In this preliminary study, maternal and neonatal oral swabs and neonatal fecal (meconium and transitional stool) swabs were selected from a pool of samples from healthy mother-neonate pairs participating in the pilot phase of CELSPAC: TNG during their hospital stay. The DNA was extracted and bacteriome profiles were determined by 16S rRNA amplicon sequencing (Illumina). In the final dataset, 33 mother-neonate pairs were exposed to antibiotics during C-section or vaginal delivery (cases; +IAP) and the vaginal delivery without IAP (controls, -IAP) took place in 33 mother-neonate pairs. Differences in alpha diversity (Shannon index, p=0.01) and bacterial composition (PERMANOVA, p<0.05) between the +IAP and -IAP groups were detected only in neonatal oral samples collected ≤48 h after birth. No significant differences between meconium bacteriomes of the +IAP and -IAP groups were observed (p>0.05). However, the IAP was associated with decreased alpha diversity (number of amplicon sequence variants, p<0.001), decreased relative abundances of the genera Bacteroides and Bifidobacterium, and increased relative abundances of genera Enterococcus and Rothia (q<0.01 for all of them) in transitional stool samples. The findings of this study suggest that exposure to IAP may significantly influence the early development of the neonatal oral and gut microbiomes. IAP affected the neonatal oral bacteriome in the first two days after birth as well as the neonatal fecal bacteriome in transitional stool samples. In addition, it highlights the necessity for further investigation into the potential long-term health impacts on children.

• MeSH
• antibakteriální látky * aplikace a dávkování   MeSH
• antibiotická profylaxe * metody   MeSH
• Bacteria genetika   klasifikace   izolace a purifikace   účinky léků   MeSH
• císařský řez MeSH
• dospělí MeSH
• feces * mikrobiologie   MeSH
• lidé MeSH
• mekonium mikrobiologie   MeSH
• novorozenec MeSH
• RNA ribozomální 16S genetika   MeSH
• střevní mikroflóra účinky léků   MeSH
• těhotenství MeSH
• ústa * mikrobiologie   MeSH
• vedení porodu MeSH
• Check Tag
• dospělí MeSH
• lidé MeSH
• mužské pohlaví MeSH
• novorozenec MeSH
• těhotenství MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH

Genome-wide association studies (GWAS) have identified more than 200 common genetic variants independently associated with colorectal cancer (CRC) risk, but the causal variants and target genes are mostly unknown. We sought to fine-map all known CRC risk loci using GWAS data from 100,204 cases and 154,587 controls of East Asian and European ancestry. Our stepwise conditional analyses revealed 238 independent association signals of CRC risk, each with a set of credible causal variants (CCVs), of which 28 signals had a single CCV. Our cis-eQTL/mQTL and colocalization analyses using colorectal tissue-specific transcriptome and methylome data separately from 1299 and 321 individuals, along with functional genomic investigation, uncovered 136 putative CRC susceptibility genes, including 56 genes not previously reported. Analyses of single-cell RNA-seq data from colorectal tissues revealed 17 putative CRC susceptibility genes with distinct expression patterns in specific cell types. Analyses of whole exome sequencing data provided additional support for several target genes identified in this study as CRC susceptibility genes. Enrichment analyses of the 136 genes uncover pathways not previously linked to CRC risk. Our study substantially expanded association signals for CRC and provided additional insight into the biological mechanisms underlying CRC development.

• MeSH
• Asijci * genetika   MeSH
• běloši * genetika   MeSH
• celogenomová asociační studie * MeSH
• genetická predispozice k nemoci * MeSH
• jednonukleotidový polymorfismus * MeSH
• kolorektální nádory * genetika   MeSH
• lidé MeSH
• lokus kvantitativního znaku * MeSH
• mapování chromozomů MeSH
• sekvenování exomu MeSH
• studie případů a kontrol MeSH
• transkriptom MeSH
• východní Asiaté MeSH
• Check Tag
• lidé MeSH
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Research Support, N.I.H., Extramural MeSH

Patients with burn injury and inhalation injury are highly susceptible to infectious complications, including opportunistic pathogens, due to the loss of skin cover and mucosal damage of respiratory tract as well as the disruption of homeostasis. This case report, a 34-year-old man suffered critical burns, provides the first literature description of triple-impact immunoparalysis (critical burns, inhalation injury, and SARS-CoV-2 infection), leading to a lethal multifocal infection caused by several fungi including very rare environmental representatives Metschnikowia pulcherrima and Wickerhamomyces anomalus. The co-infection by these common environmental yeasts in a human is unique and has not yet been described in the literature. Importantly, our patient developed refractory septic shock and died despite targeted antifungal therapy including the most potent current antifungal agent-isavuconazole. It can be assumed that besides immunoparalysis, effectiveness of therapy by isavuconazole was impaired by the large distribution volume in this case. As this is a common situation in intensive care patients, routine monitoring of plasmatic concentration of isavuconazole can be helpful in personalization of the treatment and dose optimization. Whatmore, many fungal species often remain underdiagnosed during infectious complications, which could be prevented by implementation of new methods, such as next-generation sequencing, into clinical practice.

• MeSH
• antifungální látky * terapeutické užití   MeSH
• COVID-19 * imunologie   komplikace   MeSH
• dospělí MeSH
• fatální výsledek MeSH
• koinfekce mikrobiologie   farmakoterapie   imunologie   MeSH
• lidé MeSH
• mykózy farmakoterapie   mikrobiologie   imunologie   diagnóza   MeSH
• nitrily terapeutické užití   MeSH
• popálení komplikace   mikrobiologie   MeSH
• pyridiny terapeutické užití   MeSH
• Saccharomycetales genetika   účinky léků   imunologie   MeSH
• SARS-CoV-2 imunologie   MeSH
• sepse farmakoterapie   mikrobiologie   imunologie   MeSH
• triazoly terapeutické užití   MeSH
• Check Tag
• dospělí MeSH
• lidé MeSH
• mužské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• kazuistiky MeSH

Colorectal cancer (CRC) is a leading cause of mortality worldwide. We conducted a genome-wide association study meta-analysis of 100,204 CRC cases and 154,587 controls of European and east Asian ancestry, identifying 205 independent risk associations, of which 50 were unreported. We performed integrative genomic, transcriptomic and methylomic analyses across large bowel mucosa and other tissues. Transcriptome- and methylome-wide association studies revealed an additional 53 risk associations. We identified 155 high-confidence effector genes functionally linked to CRC risk, many of which had no previously established role in CRC. These have multiple different functions and specifically indicate that variation in normal colorectal homeostasis, proliferation, cell adhesion, migration, immunity and microbial interactions determines CRC risk. Crosstissue analyses indicated that over a third of effector genes most probably act outside the colonic mucosa. Our findings provide insights into colorectal oncogenesis and highlight potential targets across tissues for new CRC treatment and chemoprevention strategies.

• MeSH
• celogenomová asociační studie MeSH
• Evropané * genetika   MeSH
• genetická predispozice k nemoci MeSH
• jednonukleotidový polymorfismus genetika   MeSH
• kolorektální nádory * genetika   MeSH
• lidé MeSH
• multiomika MeSH
• východní Asiaté * genetika   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• metaanalýza MeSH
• práce podpořená grantem MeSH
• Research Support, N.I.H., Extramural MeSH

• Publikační typ
• tisková chyba MeSH

• MeSH
• COVID-19 * MeSH
• informace pro uživatele zdravotní péče MeSH
• komunikace MeSH
• koronavirové infekce MeSH
• lidé MeSH
• společenské stigma * MeSH
• stereotypizace MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• populární práce MeSH