Apmis最新文献

Medical writing is an art but still it is usually not in the curriculum of medical students. With the beginning of scientific activity during residency, many perceive this gap increasingly, and stay behind their own expectations in their scientific productivity. Many universities offer courses to teach scientific writing and many books and article address this void, but in real life the main work to carve a readable paper out of a pile of unsorted data remains often in the hands of the scientific supervisors. This little paper tries to address this issue with a focus on typical pathology related subjects by outlining the structure of a paper and explaining typical dos and don'ts of crafting a publishable scientific paper as a pathology resident.

Helicobacter pylori is a successful etiologic gastric agent that reaches a prevalence around 80% in Colombia. This bacterium is extremely diverse and has shown a phylogeographic pattern. The objective of this study was to perform an analysis of genomic epidemiology of H. pylori in Colombia. We enriched our set of 29 newly sequenced Colombian H. pylori genomes with additional data from public databases, reaching a total of 221 genomes in our dataset. Phylogenetic characterization was carried out using MLST and whole genome SNP analysis. We also performed a characterized the diversity of virulence factors and mutations associated with antimicrobial resistance. Phylogenetic analyzes showed two new Colombian H. pylori clades. Furthermore, many virulence genotype combinations were found, mutations associated with resistance were found for all the studied antibiotics, highlighting 14.4% of the genomes presented profiles associated with resistance to more than one family of antibiotics. Our analyzes described the genomics of Colombian H. pylori and verify the presence of a population group formed exclusively by Colombian isolates. We demonstrated the great diversity among the isolates and that the analysis by comparative genomics of H. pylori are valuable tools to assess the diversity, virulence, and resistance of H. pylori.

There is a need for standardized methods for antimicrobial susceptibility testing (AST) of anaerobic bacteria involved in oral and extra-oral infections. We tested the recently published EUCAST disk diffusion method for rapidly growing anaerobes on selected oral anaerobes. AST of 20 strains of Prevotella spp., 11 strains of Porphyromonas gingivalis, and six Fusobacterium nucleatum complex strains was performed with amoxicillin and metronidazole disks using EUCAST guidelines. Plates were incubated anaerobically, and inhibition zones were evaluated after 20 h (EUCAST recommendations) and again after 44 h. The recommended agar supported the growth of all 38 strains. Twenty-hour incubation was sufficient for the assessment of inhibition zone diameters of Fusobacterium strains. Although approved for Prevotella, an extended study of Prevotella species showed inconsistent growth within the EUCAST time limit of 20 h for some strains. All P. gingivalis strains required 44 h of incubation for the evaluation of inhibition zones. The EUCAST disk diffusion method for AST of rapidly growing anaerobes is applicable to members of the Fusobacterium nucleatum complex. P. gingivalis and several oral strains of Prevotella needed 44 h of incubation to enable reading of diffusion diameter. Further studies are necessary to validate the prolonged incubation of slow-growing anaerobes.

The role of the vitamin D receptor (VDR) in inflammatory bowel disease (IBD) is poorly described. The aim of this study was to examine the relationship between immunohistochemical VDR expression and IBD activity. The immunohistochemical expression of VDR was analysed in biopsies from active and inactive IBD in 28 patients (ulcerative colitis: 21, Crohn's disease: 7) and 12 non-IBD controls. VDR expression did not change in active compared to inactive disease (p = 0.40 in epithelium and p = 0.29 in stroma). There was a trend for higher VDR expression in controls compared to IBD patients. No relationship was found between VDR expression and histologic inflammation (r = −0.19, p = 0.89 for epithelium and r = 0.13, p = 0.35 for stroma), colonoscopic picture and clinical and laboratory measures including serum 25(OH) vitamin D status (r = −0.91, p = 0.82). IBD disease activity did not correlate to VDR immunohistochemical expression, nor did it differ compared to controls. These results partly conflict with prior studies, but these have only shown modest correlations. Prospective studies investigating VDR activity between IBD and controls should be contemplated.

The ovarian oncobiome is subject to increasing scientific focus, but a potential link between bacterial dysbiosis and ovarian carcinogenesis remains controversial. Our primary aim was to characterize the bacterial microbiota in epithelial ovarian cancer samples. Secondarily, we aimed to compare results from the bacterial microbiota in formalin-fixed, paraffin-embedded ovarian tissue samples from 194 patients with epithelial ovarian cancer, fallopian tube tissue samples from 16 patients with serous tubal intraepithelial carcinomas and in benign fallopian tube tissue samples from 25 patients. Bacterial abundance was investigated by means of 16S rDNA Next Generation Sequencing. The 16S rDNA sequencing run produced a very low number of bacterial reads. Only seven samples displayed bacterial reads above 5000. Validation of detected bacterial reads by qPCR was negative and indicative of results being background amplification. Our results indicate no amount of bacterial biomass in the ovarian cancer, benign fallopian tube and in the samples with serous tubal intraepithelial carcinomas. The findings underline the importance of validating results from bacterial microbiota studies with additional technical assays before any conclusion may be drawn.

Shotgun metagenomics offers a broad detection of pathogens for rapid blood stream infection of pathogens but struggles with often low numbers of pathogens combined with high levels of human background DNA in clinical samples. This study aimed to develop a shotgun metagenomics protocol using blood spiked with various bacteria and to assess bacterial DNA extraction efficiency with human DNA depletion. The Blood Pathogen Kit (Molzym) was used to extract DNA from EDTA-whole blood (WB) and plasma samples, using contrived blood specimens spiked with bacteria for shotgun metagenomics diagnostics via Oxford Nanopore sequencing and PCR-based library preparation. Results showed that bacterial reads were higher in WB than plasma. Differences for Staphylococcus aureus and Streptococcus pneumoniae were more pronounced compared to Escherichia coli. Plasma samples exhibited better method reproducibility, with more consistent droplet digital PCR results for human DNA. The study found that extraction was more efficient for Gram-positive bacteria than Gram-negative, suggesting that the human DNA depletion exerts a negative effect on Gram-negative bacteria. Overall, shotgun metagenomics needs further optimisation to improve bacterial DNA recovery and enhance pathogen detection sensitivity. This study highlights some critical steps in the methodology of shotgun metagenomic-based diagnosis of blood stream infections using Nanopore sequencing.