[Study of the resistome of human microbial communities using a targeted panel of antibiotic resistance genes in COVID-19 patients].

AIM: To study overall drug resistance genes (resistome) in the human gut microbiome and the changes in these genes during COVID-19 in-hospital therapy. MATERIALS AND METHODS: A single-center retrospective cohort study was conducted. Only cases with laboratory-confirmed SARS-CoV-2 RNA using polymerase chain reaction in oro-/nasopharyngeal swab samples were subject to analysis. The patients with a documented history of or current comorbidities of the hepatobiliary system, malignant neoplasms of any localization, systemic and autoimmune diseases, as well as pregnant women were excluded. Feces were collected from all study subjects for subsequent metagenomic sequencing. The final cohort was divided into two groups depending on the disease severity: mild (group 1) and severe (group 2). Within group 2, five subgroups were formed, depending on the use of antibacterial drugs (ABD): group 2A (receiving ABD), group 2AC (receiving ABD before hospitalization), group 2AD (receiving ABD during hospitalization), group 2AE (receiving ABD during and before hospitalization), group 2B (not receiving ABD). RESULTS: The median number of antibiotic resistance (ABR) genes (cumulative at all time points) was significantly higher in the group of patients treated with ABD: 81.0 (95% CI 73.8-84.5) vs. 51.0 (95% CI 31.1-68.4). In the group of patients treated with ABD (2A), the average number of multidrug resistance genes (efflux systems) was significantly higher than in controls (group 2B): 47.0 (95% CI 46.0-51.2) vs. 21.5 (95% CI 7.0-43.9). Patients with severe coronavirus infection tended to have a higher median number of ABR genes but without statistical significance. Patients in the severe COVID-19 group who did not receive ABD before and during hospitalization also had more resistance genes than the patients in the comparison group. CONCLUSION: This study demonstrated that fewer ABR genes were identified in the group with a milder disease than in the group with a more severe disease associated with more ABR genes, with the following five being the most common: SULI, MSRC, ACRE, EFMA, SAT.

[Frequency and risk factors for thromboembolic complications in patients with inflammatory bowel diseases].

BACKGROUND: Inflammatory bowel diseases (IBD) are characterized by chronic immune inflammation of the mucous membrane and/or the thickness of the intestinal wall, and are also accompanied by disorders of the blood clotting system and the development of a hypercoagulation state. AIM: To identify the frequency of thromboembolic complications (TEC) in IBD patients and to determine the influence of acquired and inherited hypercoagulation factors that contribute to the development of TEС. MATERIALS AND METHODS: The clinical status of 1,238 IBD patients who were treated in 2019 was evaluated. Of these, 748 patients with ulcerative colitis (UC) and 490 patients with Crohn's disease (CD). Among UC patients, there were 369 (49.3%) men and 379 (50.7%) women. In 10.1% of patients with UC, there were clinically significant feasibility studies. There were 227 (46.3%) men and 263 (53.7%) women among patients with CD; 7.3% of patients with CD had clinically significant feasibility studies. RESULTS: In general 112 (9.0%) of 1,238 IBD patients had clinically significant feasibility studies. Among patients with UC (n=748), 76 (10.2%) showed clinically significant feasibility studies. Among patients with CD (n=490), 36 (7.3%) had a feasibility study. Of 112 IBD patients with clinically significant TEC, 45 (40.2%) had genetic polymorphisms that increase affinity for fibrinogen, increase platelet aggregation, and contribute to a decrease in the activity of folate cycle enzymes, including methylenetetrahydrofolate reductase, which may be manifested by a moderate increase in homocysteine levels. Of the 45 IBD patients with clinically significant TEC due to inherited factors, 30 (66.6%) patients had UC, 15 (33.7%) patients had CD (hazard ratio 1.038, 95% confidence interval 0.7461.444; 2=0.049; p=0.83921); 67 (59.8%) patients with IBD who had clinically significant TEC did not have genetic polymorphisms leading to hypercoagulation. CONCLUSION: Based on the analysis, we can conclude that such risk factors for the development of TEC as the status of a smoker, long bed rest, taking hormonal contraceptives, varicose veins of the lower extremities, high activity of the disease, glucocorticoids therapy, the extent of intestinal damage in patients with IBD, genetic factors, should be taken into account by gastroenterologists in the treatment of patients with UC and CD. The hereditary factor of hypercoagulation equally affects the development of TEC, both in patients with UC and CD.

[Zonulin and I-FABP are markers of enterocyte damage in celiac disease].

AIM: To evaluate the level of serum I-FABP (Fatty-Acid-Binding Protein a protein that binds fatty acids) and fecal zonulin as markers of the permeability of the mucous membrane of the small intestine in celiac patients. MATERIALS AND METHODS: A total of 151 celiac patients (25 men and 126 women) were examined. The median age was 42 years. Group I included 58 patients with newly diagnosed celiac disease; in group 2 38 patients, knowingly or unknowingly violating the gluten-free diet; group 3 consisted of 55 patients strictly observing gluten-free diet. The control group consisted of 20 healthy volunteers: 4 men and 16 women. All patients underwent esophagogastroduodenoscopy by biopsy of the mucous membrane of the small intestine and assessment of duodenobioptates according to Marsh. In the blood serum, the level of antibodies to tissue transglutaminase IgA and IgG was determined by the enzyme-linked immunosorbent assay using kits manufactured by Orgentec Diagnostics GmbH (Germany), the concentration of I-FABP in blood serum was determined using Hycult Biotech kits (Netherlands). The content of zonulin in feces was investigated by enzyme-linked immunosorbent assay using kits from Immundiagnostik AG (Germany). Statistical analysis was performed using the Statistica 13.3 software (StatSoft Inc., USA). RESULTS: There was a significant increase in the level of antibodies to tissue transglutaminase IgA [120.0 (41.1200)] IU/ml and IgG [31.4 (5.578.9)] IU/ml in patients of group 1 compared with group 2 [IgA 9.1 (2.987.6)] and IgG [3.8 (2.219.7)] IU/ml and group 3 [IgA 1.6 (1.03.2)] and IgG [2.2 (1.152.53)] (p0.01). The level of I-FABP in blood serum in patients of group 1 averaged 2045 pg/ml, in patients in group 2 1406 pg/ml, in patients in group 3 1000 pg/ml. All patients showed a significant increase in the mean I-FABP values compared to controls (1, 2 and control p0.01, 3 and control p=0.016). In patients with Marsh grade III AC atrophy, the I-FABP level depended on the degree of damage to the mucosa and significantly differed from the control: March IIIA (median: 1310 pg/ml, interquartile range: 12121461 pg/ml), March IIIB (median: 2090 pg/ml, interquartile range: 18122322 pg/ml) as well as Marsh IIIC (median: 2058 pg/ml, interquartile range 18582678 pg/ml). The concentration of zonulin in feces in patients of group 1 averaged 111.6 pg/mg, in patients of group 2 90.5 pg/mg. In patients of group 3 50 IU/ml. The concentration of zonulin in feces increased as the degree of mucosa atrophy increased (r=0.585, p0.01). However, despite the fact that both of these markers may indicate impaired permeability, each of them indicates damage to a certain level of the intestinal barrier, which is not always associated with the degree of mucosa atrophy. CONCLUSION: Determination of serum I-FABP and fecal zonulin levels in celiac patients allows for the assessment of intestinal permeability and can serve as non-invasive markers for monitoring ongoing structural changes in the mucosa without the need for endoscopy.

[Celiac crisis in patients with celiac disease. Case report].

Celiac crisis (CC) is a rare life-threatening course of celiac disease, observed mainly in children. In adults, CK can be the first manifestation of the disease and, very rarely, a relapse that occurs in patients who do not follow the gluten-free diet (AGD). Triggers can be stress, surgery, childbirth, etc. A clinical observation of CC developed in a 49-year-old patient with previously established latent celiac disease with subtotal villous atrophy, stage Marsh III C is presented. The patient did not comply with AHD. After severe angina, she developed anorexia, diarrhea, emaciation, coagulopathy, bilateral pulmonary embolism, infarction pneumonia, and enterogenic sepsis. As a result of intensive therapy with prednisolone, Fraxiparine, antibiotics, fresh frozen plasma and strict adherence to hypertension, remission of the disease was achieved.

[Bile acids are a risk factor for colorectal cancer].

Bile acids were first considered carcinogenic in 1939. Since then, accumulated data have associated colon cell changes with high levels of bile acids as an important risk factor for developing colorectal cancer, which is more common among people who consume large amounts of dietary fat. Secondary bile acids formed under the influence of the intestinal microbiota can cause the formation of reactive forms of oxygen and nitrogen, disruption of the cell membrane, mitochondria, DNA damage, reduction of apoptosis, increased cell mutation, turning them into cancer cells. High-fat diet, intestinal microflora, bile acids are a risk factors for colorectal cancer.

[Resected bowel syndrome: clinical course and treatment options].

The current concepts of the short bowel syndrome and malabsorption after intestinal surgery are generally accepted, but do not fully reflect the patients condition, making it difficult to diagnose and treat it. AIM: The purpose of the study is to analyze the clinical course of the patients after bowel resection, to create a classification based on the variants identified to allow for a differentiated treatment and to introduce the concept of the resected bowel syndrome. MATERIALS AND METHODS: We observed 239 patients (96 men and 143 women) aged 18 to 80 who underwent intestinal resection for 1 month to 16 years (from 2002 to 2018). The 1st group included 96 patients with small bowel resection (40 men and 56 women). The 2nd group included 39 men and 58 women with small bowel resection, including the resection of the ileocecal valve and the right-hand side of the colon (n=97). The 3rd group included 17 men and 29 women with the resection of the right-hand side of the colon or colectomy (n=46). The survey included the NRS-2002 (Nutritional Risk Screening 2002) screening test to identify nutritional risk, a clinical assessment of the symptoms that occurred after the surgery, instrumental methods (esophagogastroduodenoscopy, colonoscopy with biopsy, ultrasound of the abdominal cavity organs and the kidneys, a plain radiography of the abdominal cavity organs, an X-ray examination of the small intestine and the intestinal passage), serum citrulline and short-chain fatty acids in faeces. RESULTS: Based on the analysis of the clinical symptoms and the nutritional status of the patients, a new concept is proposed the resected bowel syndrome with two variants of its progression: either with or without the development of nutritional insufficiency of three types: the dehydration type, the protein-energy insufficiency type and a mixed type. Type 1 requires the use of antimicrobials with the control of SCFA concentrations in faeces. Type 2 requires the introduction of an optimal amount of easily digestible protein to correct protein-energy deficit. The 3rd (most severe) mixed type requires prescription of a parenteral nutrition component with the control of citrulline concentration in the blood serum. CONCLUSION: The proposed concept the resected bowel syndrome makes it possible to improve its diagnosis, take into account the variants of its progression and allow for a differentiated treatment.

[Disaccharidase deficiency and functional bowel diseases]. / Disakharidaznaia nedostatochnost' i funktsional'nye zabolevaniia kishechnika.

AIM: To elucidate the role of intestinal carbohydrases (glucoamylase, maltase, sucrose, and lactase) in the etiology and pathogenesis of functional bowel diseases (FBD). SUBJECTS AND METHODS: 74 patients (36 men and 38 women) aged 18 to 50 years with FBD were examined. According to Rome IV criteria (2016), there was diarrhea-predominant irritable bowel syndrome (IBS) in 21 patients, functional diarrhea (FD) in 33, constipation-predominant IBS in 6, functional constipation (FC) in 4, and mixed IBS in 10. The activity of carbohydrases in the small intestine mucosa (SIM) was investigated by the Dahlquist method modified by Trinder in the duodenal biopsy specimens obtained during esophagogastroduodenoscopy. RESULTS: Lactase deficiency was identified in 87.8% of the patients; maltase deficiency in 48.6%; sucrose deficiency in 51.3%; and glucoamylase deficiency in 85.1%. The activity of all the investigated enzymes was reduced in 23 (31.1%) patients with FBD; deficiency of 1-3 carbohydrases was found in 47 (63.5%). Normal enzymatic activity was established in 4 (5.4%) patients. CONCLUSION: In the majority of patients with FBD, the intestinal symptoms are caused by the decreased activity of SIM carbohydrases. Therefore, disaccharidase deficiency associated with an established damaging agent (nonsteroidal anti-inflammatory drugs, antibiotics, acute intestinal infections, etc.) should be considered to be a more precise diagnosis.

[Celiac disease detection rate in gastroenterological patients].

AIM: To determine celiac disease detection rate in patients with digestive disease. SUBJECTS AND METHODS: A total of 318 gastroenterological patients admitted to be treated at the Central Research Institute of Gastroenterology in September to October 2012 were examined. The patients' age was 18 to 74 years (mean 51.5±16.4 years). Immunoglobulin A (IgA) and immunoglobulin G (IgG) anti-gliadin antibodies (AGA), IgA anti-tissue transglutaminase (anti-tTG) antibodies and IgG anti-tTG antibodies were determined. When the antibodies were elevated, esophagogastroduodenoscopy with duodenal biopsy was performed. RESULTS: Forty-one of the 318 patients were found to have higher AGA (12.9%); out of them IgA AGA were in 17 (5.35%) patients and IgG AGA were also in 17 (5.35%). Elevated levels of both antibodies (IgA AGA and IgG AGA) were seen in 7 (2.2%) patients. Overall, the detection rate of increased AGA levels was 12.9%. The antibodies were more commonly higher in patients with liver diseases (21.8%) and in those with inflammatory bowel diseases (21.6%). Both IgA anti-tTG, IgG anti-tTG and IgA AGA, IgG AGA were detected in 6 (1.9%) of the 318 patients. The diagnosis of celiac disease was verified by duodenal histological examination in 3 (0.94%) of the 318 patients. CONCLUSION: The celiac disease detection rate in gastroenterological patients was 0.94%.

[Carbohydrase activities may serve as a marker for small intestinal mucosal recovery in patients with celiac disease].

Aim. To clinically evaluate the activity of glucoamylase, maltase, saccharase, and lactase in the small intestinal mucosa (SIM) of patients with celiac disease. Subjects and methods. Twenty-nine patents with celiac disease were examined. The disease was first detected in 8 patients; in the remaining patients, it had been diagnosed 6 months to 35 years before. The diagnosis was verified by histological examinations of duodenal biopsy specimens and by determination of immunoglobulin (Ig) A and G antibodies to tissue transglutaminase (atTG) and gliadin (AGA) by an enzyme-linked immunosorbent assay (ELISA). Carbohydrase activities were estimated in the duodenal biopsy specimens, by applying the method of A. Dahlquist. Results. In the control group, the activities of glucoamylase, maltase, saccharase, and lactase averaged 598.8+184.2, 825.3+239.3, 180.2-68.1, and 53.4+16.3 ng/glucose/mg tissue min, respectively. In the patients with celiac disease, the average activities of all the examined enteric enzymes were significantly below the normal value even they had been on a gluten-free diet (GFD) for 10 years or longer. Complete SIM structural recovery (Marsh stage 0) occurred in only 7 of 18 patients who had been on a strictly GFD. Serological (atTG and AGA) tests got also negative in all the 7 patients with completely recovered SIM. Six of the latter patients continued to have abdominal bloating and borborygmus, unstable stool with a propensity for diarrhea and weakness. Each was detected to have a lower activity of one or a few enzymes. The activity of all the carbohydrases reached its normal value in only 1 patient and she felt healthy, without perceiving any food intolerance. Conclusion. The activity of membrane enzymes may serve as a marker for the degree of SIM recovery in patients with celiac disease.

[Impact of bariatric surgery on vitamin metabolisms in obese patients].

AIM: To investigate the impact of bariatric surgery (BS) on the level of vitamins in obese patients. SUBJECTS AND METHODS: One hundred obese patients (78 women and 22 men) aged 19 to 61 years were examined. Controlled gastric banding (CGB) was carried out in 20 patients (mean body mass index (BMI), 41.3 ± 8.2 kg/m2); gastric sleeve resection (GSR) in 40 patients, and gastric shunting (GS) in 40 (the mean BMI in these groups were 41.1 ± 17.8 and 45.9 ± 6.2 kg/m2, respectively). A control group consisted of 10 apparently healthy individuals (BMI, 24.9 ± 3.2 kg/m2). An enzyme immunoassay was used to determine the serum concentrations of vitamins B1, B2, B5, B6, B9, B12, C, and D, niacin, biotin, and retinol-binding protein (RBP) before and 1 year after surgery. RESULTS: All the three groups showed a considerable decrease in the levels of vitamins C, B5, B6, and D, and RBP both prior to and following BS. More than 50% of the patients who had undergone GSR had also a lower baseline niacin level. A year after CGB, GSR, and GS, the number of patents with deficiency of these vitamins remained the same or increased. The majority of patients with the same level of vitamin B2, niacin, and folic acid (FA) were observed to have its decrease a year postsurgery. The concentration of the other test vitamins was also reduced a year after all operations; however, it remained within the normal range. GS had no substantial impact on the concentrations of FA, vitamins B2, B12, B1, and biotin. The lower serum vitamin levels were not accompanied by clinical symptoms in most patients following BS. CONCLUSION: In 80% of the patients with obesity, the levels of vitamins C, B6, and D were decreased to a variable degree. After BS, there was a rise in the number of patients with low serum vitamin C, D, B6, B5, niacin, FA, and RBP concentrations, at the same time the number of patients with FA deficiency increased by more than twice. BS did not significantly affect the metabolism of vitamins B1 B2 B12, and biotin.

[THE CHANGE IN THE CONCENTRATION OF VITAMINS AFTER BARIATRIC SURGERY].

AIM: to study the concentration of vitamins in obese patients after bariatric surgery (BS). MATERIAL AND METHODS: A total of 100 obese patients aged 19 to 61 years (78 women, 22 men). 20 patients (mean body mass index (BMI)--41.3 ± 8.2 kg/m²) had adjustable gastric banding (AGB), 40 patients--sleeve gastrectomy (SG) and 40--gastric bypass (GB) (mean BMI Group 41.1 ± 17.8 kg/m², and 45.9 ± 6.2 kg/m², respectively). The control group consisted of 10 healthy subjects (BMI--24.9 ± 3.2 kg/m²). We determined the serum level of vitamins B1, B2, B5, B6, B9, B12, C, D, niacin, biotin, and retinol-binding protein (RBP) before surgery and 1 year after it. RESULTS: There was a significant reduction ofvitamins C, B6, B5, D and RBP both before and after BS. More than half of patients after SG, also had initial reduction of niacin. A year after BS (AGB, SG, GB) the number of patients with deficiency of these vitamins remained the same or increased. Most patients with normal or even elevated levels ofvitamin B2, niacin and folic acid, has been declining a year after BO. The concentration of the rest of the vitamins also significantly decreased a year after all operations, but their performance remained within normal limits. CONCLUSION: Levels of vitamins C, B6 and D were reduced in more than 70% patients with obesity. The number of patients with low levels of vitamin C D, B6, B5, niacin, folic acid and RBP increased after BS. BS does not have a significant effect on the metabolism ofvitamins B1, B2, B12 and biotin.

[Significance of a method for determination of deamidated gliadin peptide in the diagnosis of celiac disease].

AIM: To define the value of a new enzyme immunoassay in determining the level of anti-deamidated gliadin peptide (DGP) antibodies (Abs) in the diagnosis of celiac disease. SUBJECTS AND METHODS: One hundred and twenty-four patients treated at the Department of Intestinal Pathology, Central Research Institute of Gastroenterology, were examined. Enzyme-linked immunosorbent assay (ELISA) was employed to determine Abs to tissue transglutaminase (tTG) and DGP of the IgA and IgG classes in the sera of all the patients. The diagnosis of celiac disease was verified by the histological examination of small bowel mucosa biopsy specimens. RESULTS: The examinees were divided into 3 groups: 1) 27 patients first diagnosed with celiac disease; 2) 40 patients keeping a gluten-free diet (GFD); 3) 57 patients with other gastrointestinal diseases (a comparison group). In the patients first diagnosed with celiac disease, the detection rate of elevated titers of anti-tTG and anti-DGP Abs in the IgA class was equal and constituted 92.5%; that in the IgG class was 96.2 and 55.5%, respectively. The comparison group showed an increase in the DGP levels in the IgA and IgG classes in 4 (7%) patients and a rise in tTG concentrations in the IgA and IgG classes was seen in only 2 (3.5%) patients. CONCLUSION: In the patents first diagnosed with celiac disease, the detection rate of elevated levels of anti-DGP Abs in the IgA and IgG classes is 92.5 and 96.2%, respectively, and significantly indifferent from that of IgA and IgG anti-tTG Abs. The patients keeping GFD displayed a reduction in anti-DGP Abs. The high detection rate of IgA anti-DGP Abs in the patients first diagnosed with celiac disease allows this method to be recommended for immunological diagnosis of this disease in adults.

[Whipple's disease in a patient with infantile cerebral palsy and hepatitis B virus cirrhosis].

The paper describes a patient who has developed Whipple's disease in the presence of infantile cerebral palsy and hepatitis B virus cirrhosis. After 5-year treatment with co-trimoxazole (480 mg b.i.d.), the clinical manifestations subsided and PAS-positive macrophages were no longer detectable in the small intestinal mucosal biopsy specimens. Subsequent worsening of the patient's condition was associated with the progression of liver cirrhosis.