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According to numerous studies, gut microbiota plays a significant role in multiple sclerosis (MS) development. However, data on changes in the gut microbiota in MS is often contradictory. The most common approach in gut microbiota research is the 16S ribosomal RNA sequencing of fecal microbiota. However, such data do not reflect the composition of the entire body microbiota. There is also a lack of data on microbiota markers in the cerebrospinal fluid (CSF) of patients with MS and predisposing conditions.Objective: to assess the level of microbial markers in the CSF of patients with MS and radiologically isolated syndrome (RIS).Patients and methods. We used gas chromatography-mass spectrometry (GC-MS) to evaluate microbial markers levels in eight patients with MS, five patients with RIS, and seven controls.Results and discussion. We found an increase in microbial load in patients with MS, indicating a possible association of MS with polymicrobial infection. In particular, an increase in the content of Streptococcus markers was observed, as well as a tendency to a three-fold increase in the campesterol content (a marker of campesterol-producing microfungi) in the CSF of patients with MS, compared to the control group (diagnostic punctures, various diseases of the nervous system of a non-autoimmune or inflammatory nature, not acute states).Conclusion. GC-MS of microbial markers can be used to assess the presence of microbial markers in the CSF. The CSF of patients with MS contains an increased amount of various microbial markers, which may indicate a possible association of MS with polymicrobial infection.

The relationship between type 2 diabetes mellitus (T2DM) and oral microbiota is still insufficiently recognized. In the present study, we compared the salivary microbiome of nondiabetic individuals, treatment-naïve diabetic patients, and diabetic patients treated with metformin or a combination of insulin and other drugs. The α- and β-diversity demonstrated significant differences in the salivary microbiome between the nondiabetic people and patients with a history of diabetes, while little divergence was found among individuals with a history of diabetes. After characterizing the effects of periodontitis on the microbial composition of each group, the salivary microbiome of the treatment-naïve diabetic patient group was compared with that of nondiabetic people and the metformin/combined treatment groups. The results revealed changes in the contents of certain bacteria after both the onset and the treatment of diabetes; among these differential bacteria, Blautia_wexlerae, Lactobacillus_fermentum, Nocardia_coeliaca and Selenomonas_artemidis varied in all processes. A subsequent correlational analysis of the differential bacteria and clinical characteristics demonstrated that salivary microbes were related to drug treatment and certain pathological changes. Finally, the four common differential bacteria were employed for distinguishing the treatment-naïve diabetic patients from the nondiabetic people and the treated patients, with prediction accuracies of 83.3%, 75% and 75%, respectively.

Autoimmune hepatitis (AIH) is a common cause of liver cirrhosis. To identify the characteristics of the oral microbiome in patients with AIH, we collected 204 saliva samples including 68 AIH patients and 136 healthy controls and performed microbial MiSeq sequencing after screening. All samples were randomly divided into discovery cohorts (46 AIH and 92 HCs) and validation cohorts (22 AIH and 44 HCs). Moreover, we collected samples of 12 AIH patients from Hangzhou for cross-regional validation. We described the oral microbiome characteristics of AIH patients and established a diagnostic model. In the AIH group, the oral microbiome diversity was significantly increased. The microbial communities remarkably differed between the two groups. Seven genera, mainly Fusobacterium, Actinomyces and Capnocytophaga, were dominant in the HC group, while 51 genera, Streptococcus, Veillonella and Leptotrichia, were enriched in the AIH group. Notably, we found 23 gene functions, including Membrane Transport, Carbohydrate Metabolism, and Glycerolipid metabolism that were dominant in AIH and 31 gene functions that prevailed in HCs. We further investigated the correlation between the oral microbiome and clinical parameters. The optimal 5 microbial markers were figured out through a random forest model, and the distinguishing potential achieved 99.88% between 46 AIH and 92 HCs in the discovery cohort and 100% in the validation cohort. Importantly, the distinguishing potential reached 95.55% in the cross-regional validation cohort. In conclusion, this study is the first to characterize the oral microbiome in AIH patients and to report the successful establishment of a diagnostic model and the cross-regional validation of microbial markers for AIH. Importantly, oral microbiota-targeted biomarkers may be able to serve as powerful and noninvasive diagnostic tools for AIH.

Представлен опыт применения метода газовой хроматографии с масс-селективной детекцией ионов (ГХ-МС) для определения маркеров потенциальных возбудителей в качестве метода этиологической диагностики при гнойно-воспалительных заболеваниях кисти. В группу исследования были включены 63 пациента с указанной патологией кисти. Метод ГХ-МС позволил выявить присутствие в исследуемом биологическом материале (раневое отделяемое, капиллярная кровь) бактериальные маркеры представителей кишечной микрофлоры, что подтверждает представление о транслокации бактериальных токсинов и микроорганизмов в очаг воспаления. Определены маркеры трудно культивируемых, посредством классических культуральных бактериологических методов исследований, облигатных анаэробов представителей эндогенной микробиоты., The article presents the experience of using the method of gas chromatography with mass-selective detection of ions (GC-MS) to determine markers of potential pathogens as a method of etiological diagnosis in purulent-inflammatory diseases of the hand. The study group included 63 patients with the indicated hand pathology. The GC-MS method revealed the presence of bacterial markers of representatives of the intestinal microflora in the biological material under study (wound separable, capillary blood), which confirms the idea of translocation of bacterial toxins and microorganisms into the focus of inflammation. Markers of difficult-to-cultivate, through classical culture bacteriological research methods, obligate anaerobes of endogenous microbiota representatives have been identified., Вестник Национального медико-хирургического Центра им. Н.И. Пирогова, Выпуск 4 2021, Pages 52-55

Gut microbiota make up the largest microecosystem in the human body and are closely related to chronic metabolic diseases. Herein, 520 fecal samples are collected from different regions of China, the gut microbiome in chronic kidney disease (CKD) is characterized, and CKD classifiers based on microbial markers are constructed. Compared with healthy controls (HC, n = 210), gut microbial diversity is significantly decreased in CKD (n = 110), and the microbial community is remarkably distinguished from HC. Genera Klebsiella and Enterobacteriaceae are enriched, while Blautia and Roseburia are reduced in CKD. Fifty predicted microbial functions including tryptophan and phenylalanine metabolisms increase, while 36 functions including arginine and proline metabolisms decrease in CKD. Notably, five optimal microbial markers are identified using the random forest model. The area under the curve (AUC) reaches 0.9887 in the discovery cohort and 0.9512 in the validation cohort (49 CKD vs 63 HC). Importantly, the AUC reaches 0.8986 in the extra diagnosis cohort from Hangzhou. Moreover, Thalassospira and Akkermansia are increased with CKD progression. Thirteen operational taxonomy units are correlated with six clinical indicators of CKD. In conclusion, this study comprehensively characterizes gut microbiome in non‐dialysis CKD and demonstrates the potential of microbial markers as non‐invasive diagnostic tools for CKD in different regions of China., Compared with healthy controls, gut microbial diversity in CKD is significantly reduced, Klebsiella and Akkermansia are significantly increased, Roseburia and Faecalibacterium are significantly reduced, and the predictive function of gut microbiota such as ascorbate metabolism and lipopolysaccharide biosynthesis is significantly enhanced. Akkermansia increases along with the progression of CKD, which is positively correlated with serum creatinine and blood urea nitrogen, and negatively correlated with estimated glomerular filtration rate, and could be used as a therapeutic target to improve the prognosis of CKD. Importantly, gut microbial markers have strong diagnostic potential for CKD and achieve cross‐regional validation, which can be used as a non‐invasive diagnostic tool for CKD.

Background and Purpose: Early life microbiota plays a crucial role in human health by acting as a barrier from pathogens' invasion and maintaining the intestinal immune homoeostasis. Altered fecal microbiota (FM) ecology was reported in newborns affected by intestinal ischemia. Our purpose was to describe, in these patients, the FM, the mucosal microbiota (MM) and the mucosal immunity. Methods: Fourteen newborns underwent intestinal resection because of intestinal ischemia. FM and MM were determined through targeted-metagenomics, diversity assignment and Kruskal-Wallis analyses of Operational taxonomic units (OTUs). The mucosal immune cells were analyzed through cytofluorimetry. Results and Conclusion: Based on the severity intestinal injueris we identified two groups: extensive (EII) and focal intestinal ischemia (FII). FM and MM varied in EII and FII groups, showing in the EII group the predominance of Proteobacteria and Enterobacteriaceae and the reduction of Bacteroidetes and Verrucomicrobia for both microbiota. The MM was characterized by a statistically significant reduction of Bacteroides, Lachnospiraceae and Ruminococcaceae and by a higher diversity in the EII compared to FII group. FM showed a prevalence of Proteobacteria, while the Shannon index was lower in the EII compared to FII group. An overall increment in B- and T-lymphocytes and Natural killer (NK) T-like cells was found for EII mucosal samples associated to an increment of TNF-α and INF-γ expressing cells, compared to FII group. FM and MM carry specific signatures of intestinal ischemic lesions. Further research may be crucial to address the role of specific taxa in EII, expecially with reference to inflammation grade and ischemia extension.

A method for identifying mixed infections, dysbioses, and inflammatory processes from specific markers (fatty acids, aldehydes, and sterols) by chromatographic mass spectrometry was used to study the microecological status of patients eligible for liver transplantation. Heterogeneous microorganisms (aerobes, anaerobes, actinobacteria, fungi, and viruses) were quantified from molecular markers in an experiment within 3 hours after the samples were sent to the laboratory. The examinees were found to have excessive growth of gram-positive anaerobes (Clostridia, eubacteria), actinobacteria of the genera Streptomyces, Nocardia, and Candida yeasts. Next were staphylococci, streptococci, and gram-negative microorganisms of the group Moraxella/Acinetobacter, Pseudomonas aeruginosa, Helicobacter pylory, and Fusobacterium/Haemophylus. Antibiotics and probiotics were chosen to correct dysbiosis and infection, by taking into account the data available in the literature. The speed and accuracy of the procedure ensure real-time monitoring of a treatment process under control of mass spectrometry.

Aiming to identify factors causing the adverse health effects associated with moisture damaged indoor environments, we analyzed immunotoxicological potential of settled dust from moisture damaged and reference schools in relation to their microbiological composition. Mouse RAW264.7 macrophages were exposed to settled dust samples (n= 25) collected from moisture damaged and reference schools in Spain, The Netherlands and Finland. After exposure, we analyzed production of inflammatory markers [nitric oxide (NO), tumor necrosis factor (TNF)α, interleukin (IL)-6 and macrophage inflammatory protein (MIP)2] as well as mitochondrial activity, viability, apoptosis and cell cycle arrest. Furthermore, particle counts, concentration of selected microbial groups as well as chemical markers ergosterol, 3-hydroxy fatty acids, muramic acid, endotoxins and glucans were measured as markers of exposure. Dust from moisture damaged schools in Spain and The Netherlands induced stronger immunotoxicological responses compared to samples from reference schools; the responses to Finnish samples were generally lower with no difference between the schools. In multivariate analysis, IL-6 and apoptosis responses were most strongly associated with moisture status of the school. The measured responses correlated with several microbial markers and numbers of particles, but the most important predictor of the immunotoxicological potential of settled dust was muramic acid concentration, a marker of Gram positive bacteria. This article is protected by copyright. All rights reserved.