RESUMEN
BACKGROUND: Vaginal lactobacilli offer protection against recurrent urinary infections, bacterial vaginosis, and vaginal candidiasis. OBJECTIVE: To characterise the isolated vaginal lactobacilli strains for their probiotic properties and to compare their probiotic potential. METHODS: The Lactobacillus strains were isolated from vaginal samples by conventional culturing and identified by sequencing of the 16S rDNA fragment. Several functional properties were detected (production of hydrogen peroxide and lactic acid; antagonistic activity against Escherichia coli, Candida albicans, Candida glabrata, and Gardnerella vaginalis; auto-aggregation and adhesiveness) as well as safety (haemolytic activity, antibiotic susceptibility, presence of transferrable resistance genes). RESULTS: A total of 135 vaginal lactobacilli strains of three species, Lactobacillus crispatus (56%), Lactobacillus jensenii (26%), and Lactobacillus gasseri (18%) were characterised using several functional and safety tests. Most of L. crispatus (89%) and L. jensenii (86%) strains produced H2O2. The best lactic acid producers were L. gasseri (18.2±2.2 mg/ml) compared to L. crispatus (15.6±2.8 mg/ml) and L. jensenii (11.6±2.6 mg/ml) (p<0.0001; p<0.0001, respectively). L. crispatus strains showed significantly higher anti-E. coli activity compared to L. jensenii. L. gasseri strains expressed significantly lower anticandidal activity compared to L. crispatus and L. jensenii (p<0.0001). There was no significant difference between the species in antagonistic activity against G. vaginalis. Nearly a third of the strains were able to auto-aggregate while all the tested strains showed a good ability to adhere to HeLa cells. None of the tested lactobacilli caused haemolysis. Although phenotypical resistance was not found to ampicillin, chloramphenicol, erythromycin, gentamycin, tetracycline, and vancomycin, the erm(B), tet(M), and tet(K) were detected in some strains. All strains were resistant to metronidazole, trimethoprim/sulfamethoxazole, and kanamycin. CONCLUSIONS: Our study revealed that the production of different antimicrobial metabolites is highly strain-specific and that the metabolites are not correlated with each other. L. crispatus displays better antagonistic activity against E. coli and Candida spp. than L. gasseri and L. jensenii; therefore; a potential probiotic candidate could be found among L. crispatus strains.
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The diagnostic value of stromal changes in carcinomas, including prostate, is under debate; in terms of limited sample tissue of biopsy, in addition to glandular alterations, the stromal changes could have additional diagnostic value, but the results in clinical settings are controversial. The research aims to evaluate the potential of stromal changes as a supplementary tool to predict the presence of higher grade carcinomas in the prostate using Masson's trichrome and Fanconi anemia complementation group M (FANCM) antibody stainings. 385 biopsies and corresponding radical prostatectomy specimens were analyzed to evaluate the rates of the diversity of ISUP grades. Of 128 upgraded prostatectomy cases, 82 were diagnosed with ISUP Gleason Grade 1 (GG1) in a biopsy. All 82 cancerous samples were stained with Masson's trichrome and FACNM antibody and compared with 82 samples without cancer to see if there was a difference in stromal composition. Additionally, 50 GG1 samples without the upgrade were stained to demonstrate if stromal changes can predict less differentiated carcinomas in the prostate. In FANCM stained samples, the average percentage of positively staining stroma over the total in non-upgraded GG1 biopsies was 36 % (13-59 %, SDâ¯=â¯11); 34 % (9-58, SDâ¯=â¯13) in samples from the upgraded cancerous group, and 44 % (22-69, SDâ¯=â¯11) in samples without cancer. In Masson's trichrome stained samples, with collagen quantified, the percentage in non-upgraded GG1 biopsies was 41 % (20-78 %, SDâ¯=â¯11); 44 % (23-89, SDâ¯=â¯15) in samples from upgraded cancerous group and 37 % (15-57, SDâ¯=â¯9) in samples without cancer. In both FANCM and Masson's trichrome, no statistical significance was found between upgraded and non-upgraded groups (pâ¯=â¯0.84 and pâ¯=â¯0.5, respectively), although some upgrades from GG1 to GG4 showed extreme values. The statistical significance was found in cancerous vs. benign samples with both FANCM (pâ¯<â¯0.01) and Masson's trichrome (pâ¯=â¯0.012). The main limiting factor is a significant overlap in staining intensity between cancerous and cancer-free groups.
Asunto(s)
Adenocarcinoma/cirugía , Biopsia , Neoplasias de la Próstata/diagnóstico , Neoplasias de la Próstata/patología , Biopsia/métodos , Humanos , Masculino , Persona de Mediana Edad , Clasificación del Tumor , Próstata/patología , Prostatectomía , Neoplasias de la Próstata/cirugía , Coloración y EtiquetadoRESUMEN
Pathadin (https://gitlab.com/Digipathology/Pathadin) was designed as a WSI oriented open-source set of tools for beginners to experience the advantages of computer-assisted image analysis and cover essential features, frequently strenuous to access with the alternative programs. It is mainly oriented to work with histology slides but also includes a significant part of modern image formats. Introducing Pathadin, the manuscript aims to improve understanding of contemporary image analysis components, resolve technophobia and misbeliefs in the computational field, simplifying pathology research work, and shifting it into a quantitative paradigm. Despite being easy to use, Pathadin includes both basic and advanced analytical algorithms, as the application of machine learning. The functionality of Pathadin is demonstrated by AI-enhanced quantification of epithelial and stromal changes in prostate carcinoma, and their dependence on ISUP grade. The material included 5 radical prostatectomy samples for training and 83 (including 11 autopsies) samples for analysis. The analytical material was stained with Masson's trichrome and Ki67, as widely available and potentially prognostic markers. An integrated solution by HistomicsTK for Ki67 evaluation was used. A U-net model for separating glands and stroma was trained, simplifying the independent analysis of these components. During the process, the model successfully highlighted glands and stroma. Masson's trichrome stain demonstrated a gradual increase in collagen expression, being statistically significant between controls vs. G1, and G3 vs. G4. Although there was considerable overlap between adjacent groups, there was only a minor overlap in collagen amount between high- and low-grade carcinomas, affirming that with further research, stroma could be an additional diagnostic marker in prostate adenocarcinoma. Ki67 showed a statistically significant gradual increase in all groups except G1 vs. G2 and G4 vs. G5. Pathadin demonstrates that there is no need for significant resources to experience the advantages of modern computer-assisted analysis.