Antimicrobial activity of dietary supplements based on bacterial lysate of Lactobacillus rhamnosus DV.

Introduction: According to WHO, antibiotic resistance is increasing to hazardous levels worldwide. Candidiasis often occurs after taking antibiotics. Therefore, antibiotic resistance is a global problem and searching for antibacterial agents is necessary. Aim: To determine the antimicrobial activity of bacterial lysate of Lactobacillus (L.) rhamnosus DV separately and with plant extracts against bacterial and yeast test cultures. Material and methods: Antimicrobial activity of Del-Immune V® (cell wall and DNA fragments from a L. rhamnosus DV) separately and with cinnamon, beetroot, and blackcurrant extracts was determined by the minimum inhibitory concentration (MIC). Twofold serial dilutions determined the MIC in previously prepared meat-peptone broth (MPB) for bacteria and liquid wort for yeast. In the study, gram-negative (Escherichia coli IEM-1, Proteus vulgaris PА-12, Pseudomonas sp. MI-2, L. rhamnosus 13/2) and gram-positive (Bacillus (B.) subtilis BТ-2, Staphylococcus aureus BМС-1) bacteria, as well as yeast (Candida (C.) albicans D-6, C. tropicalis PE-2, C. utilis BVS-65) were used as test cultures. Results: The MIC for the studied bacterial test cultures after application of L. rhamnosus DV bacterial lysates was from 1.0 ± 0.05 mg/mL to 12.5 ± 0.63 mg/mL, which was significantly less than that of the thermally inactivated control (MIC from 125.0 ± 6.25 mg/mL to 250.0 ± 12.5 mg/mL). B. subtilis BT-2 culture was the least sensitive to the action of the bacterial lysate (MIC-12.5 ± 0.63 mg/mL). It showed the best antibacterial and antifungal effect bacterial lysate with the phytonutrient blackcurrant. Conclusions: It was demonstrated that bacterial lysate of lactic acid bacteria L. rhamnosus DV exhibits antibacterial and antifungal properties during direct contact with pathogenic agents.

Circulating levels of inflammatory cytokines and angiogenesis-related growth factors in patients with osteoarthritis after COVID-19.

Background: The disease COVID-19, caused by SARS-CoV-2 infection, has a systemic effect and is associated with a number of pathophysiological mechanisms that mobilize a wide range of biomolecules. Cytokines and growth factors (GFs) are critical regulators of tissue damage or repair in osteoarthritis (OA) and are being recognized as key players in the pathogenesis of COVID-19. A clear understanding of the long-term consequences of SARS-CoV-2 infection, especially in patients with concomitant chronic diseases, is limited and needs to be elucidated. The study aimed to evaluate the degree of inflammation and levels of pro-angiogenic and hypoxic factors, as well as heat shock proteins HSP60 and HSP70 in plasma, of patients with OA after recovery from COVID-19. Methods: The research involved patients of an orthopedic specialty clinic aged 39 to 80 diagnosed with knee OA. All examined patients were divided into three groups: the Control group included conditionally healthy donors, group OA included patients with knee OA mainly stage II or III and the group of OA and COVID-19 included patients with OA who had COVID-19. The plasma levels of pro-inflammatory molecules IL-1ß, IL-6, TNF-α, NF-κB, angiogenic factors VEGF, FGF-2, PDGF, hypoxic factor HIF-1α and molecular chaperones HSP60 and HSP70 were measured by enzyme-linked immunosorbent assay. Results: The study showed that in both groups of patients, with OA and convalescent COVID-19, there was an increase in the plasma level of IL-1ß and a decrease in TNF-α and NF-κB levels when compared to healthy controls. Systemic deregulation of the cytokine profile was accompanied by reduction in plasma levels of pro-angiogenic growth factors, most pronounced in cases of VEGF and PDGF. This analysis did not reveal any significant difference in the plasma level of HIF-1α. A decrease in the level of stress protein HSP60 in the blood of patients with OA, as well as those patients who have had SARS-CoV-2 infection, has been established. Conclusion: The results suggest the potential role pro-inflammatory cytokines and angiogenesis-related growth factors in pathogenesis of both joint pathologies and long-term systemic post-COVID-19 disorders.

Nutrition program, physical activity and gut microbiota modulation: a randomized controlled trial to promote a healthy lifestyle in students with vitamin D3 deficiency.

BACKGROUND: University students are a high-risk group for stress, consumption of junk food and significant weight gain over a short period. Inadequate vitamin D intake has been linked to many health issues, including chronic headache, apathy, aggression and depression. Furthermore, vitamin D deficiency led to dysbiosis of the gut microbiota. The purpose of our study was to estimate the effect of 90-days healthy lifestyle programs along with gut microbiota modulation in university students with vitamin D3 deficiency. METHODS: In this randomized controlled trial, 130 students (18-25 years old) with vitamin D deficiency were recruited. Both the standard care group (N.=65) and the intervention group (N.=65) received a 3-months course of individually selected nutrition program and physical activity (8000-10,000 steps daily). The intervention group received an additional treatment with synbiotic Acidolac and vitamin D3 for 3 months. The psycho-emotional status of the participants was assessed by a validated questionnaire that examined situational anxiety. In all students, blood pressure, anthropometric variables, as well as laboratory metabolic parameters, were recorded. RESULTS: In both groups, vitamin D3 deficiency was associated with instability and lability of mental processes, mood swings, bad sleep, high rates of stuck and agitation for any problem. Combined therapy (diet, physical activity and synbiotic) induced a significant improvement in the psycho-emotional state of students. The 90-days therapy vitamin D3 increased the level of vitamin D3 in serum in the intervention group. Lastly, we observed a decrease in the body weight, body mass index, waist circumference and fatty mass, only in students included in the interventional group. CONCLUSIONS: Nutrition program, physical activity, vitamin D3 intake and gut microbiota modulation led to both the improvement in vitamin D levels in serum and emotional harmonization.

Efficacy and safety of fecal microbiota transplantation via colonoscopy as add-on therapy in patients with mild-to-moderate ulcerative colitis: A randomized clinical trial.

Introduction: Growing evidence supports the effectiveness of fecal microbiota transplantation (FMT) in treating ulcerative colitis (UC), although its effects seem to depend on the method of introduction, the number of procedures, the donor material, and the severity of UC. Aim: This study aimed to assess FMT's clinical and microbiological efficacy, tolerability, and safety in patients with mild-to-moderate UC. Material and methods: Patients with mild-to-moderate UC were randomized into two groups. The first group (standard-care, n = 27) was treated with basic therapy-mesalazine-at a daily dose of 3 g (2 g orally + 1 g rectally). In the second group (FMT group, n = 26), while taking mesalazine at the indicated dose, each patient with UC as add-on therapy underwent a single FMT procedure with fresh material delivered by colonoscopy from a healthy donor. The clinical efficacy of treatment in both groups was evaluated after 4 and 8 weeks. The primary outcome was remission of UC, defined as a partial Mayo score ≤2, and decreased fecal calprotectin. All patients underwent bacteriological examination of feces for quantitative microbiota composition changes. Results: Clinical response in the form of a significant decrease in stool frequency and a tendency to normalize its consistency after 4 weeks was detected in 14 (51.9%) patients of the standard care group and 16 patients (61.5%) of the FMT group (p = 0.583). The Mayo score in the standard care group was 3.59 ± 1.21 and in the FMT group-3.15±1.04 (p=0.166). After 8 weeks, the main primary endpoint was achieved in 70.4% of the standard-care group patients as compared to 84.6% of participants who received FMT as add-on therapy (p = 0.215). A more pronounced decrease in Mayo score was observed in the FMT group compared to the standard-care group (1.34 ± 1.44 vs. 2.14 ± 1.4; p = 0.045). All patients also showed a significant decrease in fecal calprotectin levels, which correlated with clinical data, stool frequency, and clinical remission. An improvement in gut microbiota composition was noted in both groups, albeit it was significantly more pronounced in the FMT group. Conclusions: FTM in patients with mild-to-moderate UC is a well-tolerated, effective, and safe method of treatment in comparison to basic therapy. Clinical trial registration: https://clinicaltrials.gov/ct2/show/NCT05538026?term=kobyliak&draw=2&rank=4, identifier: NCT05538026.

Mechanisms of the Impact of Hashimoto Thyroiditis on Papillary Thyroid Carcinoma Progression: Relationship with the Tumor Immune Microenvironment.

BACKGROUND: The relationship between Hashimoto thyroiditis (HT) and papillary thyroid carcinoma (PTC) remains uncertain. We assessed the impact of HT on the tumor immune microenvironment (TIME) in PTC. METHODS: Thirty patients with PTC (group 1) and 30 patients with PTC and HT (group 2) were enrolled in this pilot study. The distribution and number of CD8+ lymphocytes, plasma cells (CD138+), regulatory T cells (forkhead box P3 [FOXP3+)], mast cell tryptase (MCT+), and M2 macrophages (CD163+) were evaluated. To test the hypothesis that HT impacts PTC development via signal transducer and activator of transcription 6 (STAT6) activation and M2 macrophage polarization, we investigated STAT6 expression in tumor and stromal cells. We also evaluated vascular endothelial growth factor (VEGF) expression by lymph node metastasis (LNM) status. RESULTS: TIME showed significant between-group differences. Group 1 patients demonstrated immune desert or immune-excluded immunophenotypes, while an inflamed phenotype with more CD8+ cells (P<0.001) predominated in group 2. Immune-excluded TIME was associated with the highest LNM rate. In PTC, LNM was associated with more numerous CD163+ cells. Moreover, LNM in group 1 was associated with increased numbers of mast cells peritumorally and FOXP3+ cells intratumorally and peritumorally. Group 2 demonstrated higher STAT6 but not higher VEGF expression in tumor cells. High VEGF expression was associated with LNM regardless of HT status. CONCLUSION: Concomitant HT impacted PTC signaling via STAT6 and TIME by increasing the number of CD8+ cells. LNM is associated with increases in CD163+ cells and VEGF expression in PTC, whereas HT affected LNM through different mechanisms.

New insights on strain-specific impacts of probiotics on insulin resistance: evidence from animal study.

BACKGROUND AND AIMS: сomparative animal study of effectiveness of intermittent administration of lyophilized single-, three- and alive multistrain probiotic in short courses on insulin resistance (IR) in rats with experimental obesity. METHODS: 70 rats were divided into 7 groups (n = 10 in each). Rats of group I were left intact. Newborn rats in groups II-VII were administered monosodium glutamate (MSG) (4 mg/g) by injection. Rats in group II (MSG-obesity group) were left untreated. The rats in groups III-V received lyophilized mono-probiotics B.animalis VKL, B.animalis VKB, L.casei IMVB-7280 respectively. The rats in group VI received all three of these probiotic strains mixed together. Group VII was treated with multi-probiotic "Symbiter", containing 14 different live probiotic strains (Lactobacillus, Bifidobacterium, Propionibacterium, Acetobacter genera). RESULTS: Treatment of newborn rats with MSG lead to the development of obesity in all MSG-obesity rats and up to 20-70% after probiotic administration. Additions to probiotic composition, with preference to alive strains (group VII), led to significantly lower rates of obesity, decrease in HOMA-IR (p < 0.001), proinflammatory cytokines levels - IL-1ß (p = 0.003), IL-12Bp40 (p < 0.001) and elevation of adiponectin (p = 0.003), TGF-ß (p = 0.010) in comparison with MSG-obesity group. Analysis of results in groups treated with single-strain probiotics (groups III-V) shows significant decrease in HOMA-IR, but changes were less pronounced as compared to mixture groups and did not achieve intact rats level. Other metabolic parameters were not affected significantly by single strains. CONCLUSION: Our findings provide major clues for how to design and use probiotics with more efficient compositions in obesity and IR management and may bring new insights into how host-microbe interactions contribute to such protective effects.

The effects of Z56822977 on biosynthesis of serotonin in the brain of rats in the conditions of MSG-induced obesity.

Wstep: Badanie przeprowadzono w celu wyjasnienia wplywu Z56822977 na biosynteze serotoniny w mózgu szczurów z otyloscia wy-wolana podawaniem glutaminianu sodu (monosodium glutamate, MSG). Material i metody: W badaniu wykorzystano 18 samców szczura. Zwierzeta podzielono na trzy grupy: 1 - grupa kontrolna, 2 - grupa MSG, 3 - grupa MSG + Z56822977. Szczurzym oseskom w grupie 2 i 3 podawano podskórnie MSG rozpuszczony w soli fizjologicznej w dawce 4 mg/g masy ciala w objetosci 8 µl/g w 2., 4., 6., 8. i 10. dniu zycia. Grupie 3 podawano doustnie wodny roztwór Z56822977 w dawce 25 mg/kg w objetosci 1 ml/kg. Pierwsza dawke Z56822977 podawano po ukonczeniu 4 tygodni zycia, a nastepnie kontynuowa-no podawanie badanej substancji cyklicznie wedlug schematu tydzien podawania substancji badanej/3 tygodnie przerwy. Zwierzetom z grupy MSG podawano odpowiednio 1 ml/kg wody doustnie. Przez pierwsze 4 miesiace zycia szczury otrzymywaly standardowa karme. Zmierzono zawartosc serotoniny, tryptofanu i 5-hydroksytryptofanu (5-HTr) oraz aktywnosc hydroksylazy tryptofanowej (tryptophan hydroxylase, TRH), dekarboksylazy aminokwasów (amino acid decarboxylase, AADC) i monoaminooksydazy (MAO) w tkance mózgowej. WYNIKI: Wykazano, ze podawanie Z56822977 ma pozytywny wplyw na glówne wskazniki otylosci, co odzwierciedlaja zmiany podsta-wowych parametrów fizjologicznych i biochemicznych [zmniejszenie masy ciala o 13% vs. MSG (p < 0,05); zmniejszenie wskaznika masy ciala (body mass index, BMI), wskaznika Lee oraz masy tkanki tluszczowej trzewnej odpowiednio o 18%, 7% i 55%, (p < 0,05) w porównaniu z grupa MSG]. Zawartosc tryptofanu i serotoniny byla istotnie nizsza (p < 0,05) u szczurów z otyloscia wywolana przez MSG. W badaniach wykazano, ze u otylych szczurów aktywnosc MAO zwieksza sie o 97% (p < 0,05), a aktywnosc TRH i AADC odpowiednio o 44% i 53% (p < 0,05). Podawanie Z56822977 powodowalo zwiekszenie zawartosci serotoniny i tryptofanu w mózgach szczurów i przywracalo poziom aktywnosci enzymów (MAO, TRH, AADC) do wartosci mierzonych u zwierzat kontrolnych. WNIOSKI: Wiadomo, ze otylosc wiaze sie z zaburzeniem syntezy serotoniny w mózgu szczurów. Jednak podawanie Z56822977 prowadzi do normalizacji stezenia serotoniny i tryptofanu oraz przywrócenia prawidlowej aktywnosci enzymów uczestniczacych w biosynte-zie i degradacji serotoniny. Podawanie Z56822977, czasteczki wplywajacej na uklad serotoninergiczny, moze powodowac korzystne efekty w leczeniu otylosci wywolanej przez MSG u szczurów. Mozna rozwazac zastosowanie czasteczki Z56822977 jako nowego leku stosowanego w otylosci, jednak konieczne sa dalsze badania w celu potwierdzenia jej dzialania.

Obeticholic Acid: A New Era in the Treatment of Nonalcoholic Fatty Liver Disease.

The main treatments for patients with nonalcoholic fatty liver disease (NAFLD) are currently based on lifestyle changes, including ponderal decrease and dietary management. However, a subgroup of patients with nonalcoholic steatohepatitis (NASH), who are unable to modify their lifestyle successfully, may benefit from pharmaceutical support. Several drugs targeting pathogenic mechanisms of NAFLD have been evaluated in clinical trials for the treatment of NASH. Farnesoid X receptor (FXR) is a nuclear key regulator controlling several processes of the hepatic metabolism. NAFLD has been proven to be associated with abnormal FXR activity. Obeticholic acid (OCA) is a first-in-class selective FXR agonist with anticholestatic and hepato-protective properties. Currently, OCA is registered for the treatment of primary biliary cholangitis. However, promising effects of OCA on NASH and its metabolic features have been reported in several studies.