scholarly journals Effects of Lactobacillus salivarius LN12 in Combination with Amoxicillin and Clarithromycin on Helicobacter pylori Biofilm In Vitro

2021 ◽  
Vol 9 (8) ◽  
pp. 1611
Author(s):  
Fang Jin ◽  
Hong Yang
Keyword(s):  
Helicobacter Pylori ◽  
Inhibitory Concentration ◽  
Laser Scanning Microscopy ◽  
Confocal Laser ◽  
Clinical Strain ◽  
Dilution Method ◽  
Lactobacillus Salivarius ◽  
H Pylori ◽  
Biofilm Structure

Helicobacter pylori is a highly prevalent and harmful gastrointestinal pathogen. Antibiotic resistance and biofilm complexity have led to a decrease in the cure rate. Probiotics are considered to be an adjuvant therapy for clinical Helicobacter pylori infections. However, there is no substantial explanation for the adjuvant role of probiotics on H. pylori biofilm. In this study, the effects of probiotics in combination with amoxicillin (AMX) and clarithromycin (CLR) on H. pylori biofilms were explored in vitro for the first time. The minimum inhibitory concentration (MIC) and the fractional inhibitory concentration (FIC) for H. pylori was determined by the microbroth dilution method, and the plate counting method was used to determine the minimum biofilm removal concentration (MBEC) and survival rate for H. pylori biofilm. The biofilm structure was observed by scanning electron microscopy (SEM) and confocal laser scanning microscopy (CLSM), protein and polysaccharide contents in extracellular polymeric substances (EPS) were determined by the Bradford method and the phenol-sulfate method, respectively. The gene expression levels of cagA and vacA were evaluated by real-time qPCR. Among the ten H. pylori strains, the clinical strain 3192 showed the strongest film-forming ability, the 3192 biofilms significantly improved the resistance to AMX and CLR, and AMX and CLR showed antagonistic effects on planktonic 3192 cells. When the Lactobacillus salivarius LN12 cell-free supernatant (CFS) was in combination with AMX and CLR, the 3192 biofilm structure was destroyed to a greater extent than when separately; more biofilm biomass and protein in EPS was decreased; and the downregulation effect of the virulence gene vacA was also greater than that of single use. In this study, we suggest that the addition of LN12 to AMX and CLR may enhance the therapeutic effect of triple therapy, especially for the treatment of H. pylori biofilms.

scholarly journals In Vitro Effects of Lactobacillus plantarum LN66 and Antibiotics Used Alone or in Combination on Helicobacter pylori Mature Biofilm

2021 ◽  
Vol 9 (2) ◽  
pp. 424
Author(s):  
Jianfu Ji ◽  
Hong Yang
Keyword(s):  
Helicobacter Pylori ◽  
Antibiotic Therapy ◽  
Lactobacillus Plantarum ◽  
Laser Scanning ◽  
Extracellular Polymeric Substances ◽  
Laser Scanning Microscopy ◽  
Confocal Laser ◽  
In Vitro Effects ◽  
H Pylori

Helicobacter pylori is a gastrointestinal pathogen with high prevalence that harms human health. Studies have shown that H. pylori can form antibiotic-tolerant biofilms, which may interfere with the efficacy of clinical antibiotic therapy. Probiotics can antagonize planktonic and biofilm pathogen cells and thus may play an auxiliary role in H. pylori antibiotic therapy. However, the effects of different probiotic strains and antibiotic combinations on H. pylori biofilms need to be further investigated. We determined the cell viability of H. pylori mature biofilms after treatment with Lactobacillus plantarum LN66 cell-free supernatant (CFS), clarithromycin (CLR), and levofloxacin (LVX) alone or in combination by the XTT method. Biofilm cells were observed by scanning electron microscopy (SEM) and confocal laser scanning microscopy (CLSM). Subsequently, protein and polysaccharide concentrations in biofilm extracellular polymeric substances (EPSs) were quantitatively detected by the Bradford method and the phenol-sulfate method. The results showed that LN66 CFS had an eradication effect on mature H. pylori biofilm. When used in combination with CLR, LN66 CFS significantly attenuated the eradication effect of CLR on biofilms; in contrast, when used in combination with LVX, LN66 CFS enhanced the disrupting effect of LVX. We speculate that the different effects of CFS and antibiotic combinations on biofilms may be related to changes in the content of proteins and polysaccharides in EPS and that the combination of CFS and CLR might promote the secretion of EPS, while the combination of CFS and LVX might have the opposite effect. Accordingly, we suggest that supplementation with L. plantarum LN66 may provide additional help when therapy involving LVX is used for clinical H. pylori biofilm eradication, whereas it may impair CLR efficacy when therapy involving CLR is used.

Vacuoles Induced in Mammalian Cells by Helicobacter Cytotoxin are Acidic Organelles

1990 ◽  
Vol 48 (3) ◽  
pp. 168-169
Author(s):  
M. H. Chestnut ◽  
C. E. Catrenich
Keyword(s):  
Acridine Orange ◽  
Mammalian Cells ◽  
Laser Scanning ◽  
Laser Scanning Microscopy ◽  
Confocal Laser ◽  
Ulcer Disease ◽  
Gastroduodenal Disease ◽  
H Pylori

Helicobacter pylori is a non-invasive, Gram-negative spiral bacterium first identified in 1983, and subsequently implicated in the pathogenesis of gastroduodenal disease including gastritis and peptic ulcer disease. Cytotoxic activity, manifested by intracytoplasmic vacuolation of mammalian cells in vitro, was identified in 55% of H. pylori strains examined. The vacuoles increase in number and size during extended incubation, resulting in vacuolar and cellular degeneration after 24 h to 48 h. Vacuolation of gastric epithelial cells is also observed in vivo during infection by H. pylori. A high molecular weight, heat labile protein is believed to be responsible for vacuolation and to significantly contribute to the development of gastroduodenal disease in humans. The mechanism by which the cytotoxin exerts its effect is unknown, as is the intracellular origin of the vacuolar membrane and contents. Acridine orange is a membrane-permeant weak base that initially accumulates in low-pH compartments. We have used acridine orange accumulation in conjunction with confocal laser scanning microscopy of toxin-treated cells to begin probing the nature and origin of these vacuoles.

scholarly journals Antibacterial Effects of Grape Extracts on Helicobacter pylori

2008 ◽  
Vol 75 (3) ◽  
pp. 848-852 ◽  
Author(s):  
Joseph C. Brown ◽  
Guohui Huang ◽  
Vivian Haley-Zitlin ◽  
Xiuping Jiang
Keyword(s):  
Cell Proliferation ◽  
Helicobacter Pylori ◽  
Laser Scanning ◽  
Laser Scanning Microscopy ◽  
Confocal Laser ◽  
Antibacterial Effects ◽  
Grape Skin ◽  
Agar Dilution ◽  
H Pylori ◽  
Proliferation Assays

ABSTRACT Anti-Helicobacter pylori activities were determined by agar dilution, confocal laser scanning microscopy, and cell proliferation assays following treatment with various grape extracts. Muscadine grape skin possessed the strongest activity, followed by grape synergy (skin and seed) and seed, suggesting that higher phenolic levels do not necessarily determine overall anti-H. pylori efficacy.

scholarly journals In Silico and In Vitro Anti-Helicobacter Pylori Effects of Combinations of Phytochemicals and Antibiotics

Molecules ◽  
2019 ◽  
Vol 24 (19) ◽  
pp. 3608 ◽  
Author(s):  
Pedro Fong ◽  
Chon-Hou Hao ◽  
Chi-Cheng Io ◽  
Pou-Io Sin ◽  
Li-Rong Meng
Keyword(s):  
Helicobacter Pylori ◽  
In Silico ◽  
Inhibitory Concentration ◽  
Positive Control ◽  
Shikimate Kinase ◽  
Semialdehyde Dehydrogenase ◽  
Class 1 ◽  
H Pylori ◽  
Potential Inhibitors

Helicobacter pylori infection is a WHO class 1 carcinogenic factor of gastric adenocarcinoma. In the past decades, many studies have demonstrated the increasing trend of antibiotic resistance and pointed out the necessity of new effective treatment. This study was aimed at identifying phytochemicals that can inhibit H. pylori and possibly serve as adjuvant treatments. Here, in silico molecular docking and drug-like properties analyses were performed to identify potential inhibitors of urease, shikimate kinase and aspartate-semialdehyde dehydrogenase. These three enzymes are targets of the treatment of H. pylori. Susceptibility and synergistic testing were performed on the selected phytochemicals and the positive control antibiotic, amoxicillin. The in-silico study revealed that oroxindin, rosmarinic acid and verbascoside are inhibitors of urease, shikimate kinase and aspartate-semialdehyde dehydrogenase, respectively, in which, oroxindin has the highest potency against H. pylori, indicated by a minimum inhibitory concentration (MIC) value of 50 μg/mL. A combination of oroxindin and amoxicillin demonstrated additive effects against H. pylori, as indicated by a fractional inhibitory concentration (FIC) value of 0.75. This study identified phytochemicals that deserve further investigation for the development of adjuvant therapeutic agents to current antibiotics against H. pylori.

scholarly journals In Vitro Activities of Linezolid Alone and in Combination with Amoxicillin, Clarithromycin, and Metronidazole againstHelicobacter pylori

2000 ◽  
Vol 44 (7) ◽  
pp. 1977-1979 ◽  
Author(s):  
Alexander M. Hirschl ◽  
Petra Apfalter ◽  
Athanasios Makristathis ◽  
Manfred L. Rotter ◽  
Margit Wimmer
Keyword(s):  
Helicobacter Pylori ◽  
Inhibitory Concentration ◽  
Dilution Method ◽  
Agar Dilution Method ◽  
Fractional Inhibitory Concentration ◽  
Agar Dilution

ABSTRACT Linezolid was tested against 70 strains of Helicobacter pylori by the agar dilution method. The MIC range and MICs at which 50 and 90% of strains were inhibited were 8 to 64, 16, and 32 μg/ml, respectively. With minimum and maximum fractional inhibitory concentration summation values of 0.31 and 2.50, respectively, the combination of linezolid with amoxicillin, clarithromycin, or metronidazole showed either partial synergy or indifference for the majority of strains.

Activity of Bulgarian propolis against 94 Helicobacter pylori strains in vitro by agar-well diffusion, agar dilution and disc diffusion methods

2005 ◽  
Vol 54 (5) ◽  
pp. 481-483 ◽  
Author(s):  
Lyudmila Boyanova ◽  
Galina Gergova ◽  
Rossen Nikolov ◽  
Sirigan Derejian ◽  
Elena Lazarova ◽  
...  
Keyword(s):  
Helicobacter Pylori ◽  
Growth Inhibition ◽  
Biological Effects ◽  
Diffusion Method ◽  
Dilution Method ◽  
Agar Dilution Method ◽  
Propolis Extract ◽  
Agar Dilution ◽  
H Pylori

Propolis exhibits antimicrobial, anti-inflammatory and other biological effects. The aim of this study was to evaluate the activity of 30 % ethanolic extract of Bulgarian propolis against 94 Helicobacter pylori strains by three methods. By the agar-well diffusion method, only 13.8 % of the strains exhibited no inhibition by 30 μl propolis extract (containing 9 mg propolis) and all isolates were inhibited to some extent by 90 μl of the extract (27 mg propolis) per well. The mean diameters of growth inhibition by 30, 60 or 90 μl propolis extract or 30 μl 96 % ethanol per well were 16.8, 19.2, 27.5 and 8.3 mm, respectively. The propolis extract was more active than the ethanol (P < 0.001). With 90 μl propolis extract per well, 69.4 % of the strains exhibited large diameters of growth inhibition (⩾20 mm) versus 26.6 % with 30 μl per well (P < 0.001). With moist propolis discs, inhibition was detected in more strains (92.1 %) than with dried discs (78.2 %, P < 0.05), with mean inhibitory diameters of 18.7 and 13.8 mm, respectively. By the agar dilution method, 100 and 300 μg propolis ml−1 inhibited the growth of 57.1 % and 76.2 %, respectively, of the 21 strains tested. In conclusion, Bulgarian propolis had a strong and dose-dependent activity against most of the H. pylori strains tested. Although the effect of propolis on H. pylori in vitro is promising, further microbiological, pharmacological and clinical trials are required.

scholarly journals Antibiofilm and Antimicrobial-Enhancing Activity of Chelidonium majus and Corydalis cheilanthifolia Extracts against Multidrug-Resistant Helicobacter pylori

Pathogens ◽  
2021 ◽  
Vol 10 (8) ◽  
pp. 1033
Author(s):  
Paweł Krzyżek ◽  
Adam Junka ◽  
Wojciech Słupski ◽  
Arleta Dołowacka-Jóźwiak ◽  
Bartosz J. Płachno ◽  
...  
Keyword(s):  
Helicobacter Pylori ◽  
Drug Carrier ◽  
Multidrug Resistant ◽  
New Drugs ◽  
Clinical Strain ◽  
Antibiofilm Activity ◽  
Chelidonium Majus ◽  
Minimal Inhibitory Concentrations ◽  
H Pylori

Helicobacter pylori is a Gram-negative bacterium that colonizes the stomach of about 60% of people worldwide. The search for new drugs with activity against H. pylori is now a hotspot in the effective and safe control of this bacterium. Therefore, the aim of this research was to determine the antibacterial activity of extracts from selected plants of the Papaveraceae family against planktonic and biofilm forms of the multidrug-resistant clinical strain of H. pylori using a broad spectrum of analytical in vitro methods. It was revealed that among the tested extracts, those obtained from Corydalis cheilanthifolia and Chelidonium majus were the most active, with minimal inhibitory concentrations (MICs) of 64 µg/mL and 128 µg/mL, respectively. High concentrations of both extracts showed cytotoxicity against cell lines of human hepatic origin. Therefore, we attempted to lower their MICs through the use of a synergistic combination with synthetic antimicrobials as well as by applying cellulose as a drug carrier. Using checkerboard assays, we determined that both extracts presented synergistic interactions with amoxicillin (AMX) and 3-bromopyruvate (3-BP) (FICI = 0.5) and additive relationships with sertraline (SER) (FICI = 0.75). The antibiofilm activity of extracts and their combinations with AMX, 3-BP, or SER, was analyzed by two methods, i.e., the microcapillary overgrowth under flow conditions (the Bioflux system) and assessment of the viability of lawn biofilms after exposure to drugs released from bacterial cellulose (BC) carriers. Using both methods, we observed a several-fold decrease in the level of H. pylori biofilm, indicating the ability of the tested compounds to eradicate the microbial biofilm. The obtained results indicate that application of plant-derived extracts from the Papaveraceae family combined with synthetic antimicrobials, absorbed into organic BC carrier, may be considered a promising way of fighting biofilm-forming H. pylori.

scholarly journals In Vitro Effect of Copper (I) Complex [Cu(NN1)2](ClO4) on Vibrio harveyi BB170 Biofilm Formation

2021 ◽  
Vol 9 (11) ◽  
pp. 2273
Author(s):  
Sarita Soto-Aguilera ◽  
Brenda Modak ◽  
Maialen Aldabaldetrecu ◽  
Carla P. Lozano ◽  
Juan Guerrero ◽  
...  
Keyword(s):  
Biofilm Formation ◽  
Pathogenic Bacteria ◽  
Inhibitory Concentration ◽  
Vibrio Harveyi ◽  
Laser Scanning Microscopy ◽  
Confocal Laser ◽  
Minimal Bactericidal Concentration ◽  
Vitro Effect ◽  
Harveyi Bb170

Biofilm formation in pathogenic bacteria is an important factor of resistance to antimicrobial treatments, allowing them to survive for a long time in their hosts. In the search for new antibiofilm agents, in this work we report the activity of a copper (I) complex, [Cu(NN1)2]ClO4, synthesized with Cu (I) and NN1, an imine ligand 6-((quinolin-2-ylmethylene)amino)-2H-chromen-2-one, a derivate of natural compound coumarin. The antibacterial and antibiofilm capacity was evaluated in Vibrio harveyi BB170 used as model bacteria. Antibacterial activity was measured in vitro by minimal inhibitory concentration (MIC), minimal bactericidal concentration (MBC) and half-maximal inhibitory concentration (IC50) determination. Antibiofilm capacity of copper (I) complex was analyzed by different concentrations of IC50 values. The results showed that the sub-IC50 concentration, 12.6 µg/mL of the copper (I) complex, was able to reduce biofilm formation by more than 75%, and bacterial viability was reduced by 50%. Inverted and confocal laser scanning microscopy showed that the [Cu(NN1)2]ClO4 complex affected the biofilm structure. Therefore, the copper (I) complex is effective as an antibiofilm compound in V. harveyi BB170.

scholarly journals The Effects of Sulglycotide on the Adhesion and the Inflammation of Helicobacter Pylori

2020 ◽  
Vol 17 (8) ◽  
pp. 2918
Author(s):  
Ji Yeong Yang ◽  
Pumsoo Kim ◽  
Seok-Hoo Jeong ◽  
Seong Woong Lee ◽  
Yu Sik Myung ◽  
...  
Keyword(s):  
Helicobacter Pylori ◽  
Protein A ◽  
Enzyme Linked Immunosorbent Assay ◽  
Etiologic Factor ◽  
Dilution Method ◽  
Gastric Diseases ◽  
Ags Cells ◽  
Human Gastric Adenocarcinoma ◽  
H Pylori

Helicobacter pylori (H. pylori) is a primary etiologic factor in gastric diseases. Sulglycotide is a glycopeptide derived from pig duodenal mucin. Esterification of its carbohydrate chains with sulfate groups creates a potent gastroprotective agent used to treat various gastric diseases. We investigated the inhibitory effects of sulglycotide on adhesion and inflammation after H. pylori infection in human gastric adenocarcinoma cells (AGS cells). H. pylori reference strain 60190 (ATCC 49503) was cultured on Brucella agar supplemented with 10% bovine serum. Sulgylcotide-mediated growth inhibition of H. pylori was evaluated using the broth dilution method. Inhibition of H. pylori adhesion to AGS cells by sulglycotide was assessed using a urease assay. Effects of sulglycotide on the translocation of virulence factors was measured using western blot to detect cytotoxin-associated protein A (CagA) and vacuolating cytotoxin A (VacA) proteins. Inhibition of IL-8 secretion was measured using enzyme-linked immunosorbent assay (ELISA) to determine the effects of sulglycotide on inflammation. Sulglycotide did not inhibit the growth of H. pylori, however, after six and 12 hours of infection on AGS cells, H. pylori adhesion was significantly inhibited by approximately 60% by various concentrations of sulglycotide. Sulglycotide decreased H. pylori virulence factor (CagA and VacA) translocation to AGS cells and inhibited IL-8 secretion. Sulglycotide inhibited H. pylori adhesion and inflammation after infection of AGS cells in vitro. These results support the use of sulglycotide to treat H. pylori infections.

scholarly journals Antibacterial activity of Zn-loaded Cuban zeolite against Helicobacter pylori in comparison to its Na-loaded and unmodified counterparts

2020 ◽  
Author(s):  
Guido Cerri ◽  
Mauro Farina ◽  
Antonio Brundu ◽  
Elisabetta Gavini ◽  
Andrea Salis ◽  
...  
Keyword(s):  
Helicobacter Pylori ◽  
Direct Relationship ◽  
Cation Exchanger ◽  
Inhibitory Concentration ◽  
Large Population ◽  
Antimicrobial Properties ◽  
Original Form ◽  
Diffusion Test ◽  
H Pylori

AbstractHelicobacter pylori can be found in the stomach of about half of the humans, and a large population can be associated with serious diseases. To survive in the stomach H. pylori increases the pH locally by producing ammonia which binds to H+ becoming ammonium. This work investigated the effects on the in-vitro growth of H. pylori of a natural cation-exchanger mainly composed (≈70%) of clinoptilolite and mordenite. The zeolitized material from Cuba was evaluated in its original form (M), as well as in its Na- (M-Na) and Zn-exchanged (M-Zn) counterparts. In the preliminary agar cup diffusion test, H. pylori revealed susceptibility only to M-Zn, with a direct relationship between concentration and width of inhibition halo. Further experiments evidenced that bacterium replication increases when ammonium is supplied to the growth medium and decreases when zeolites subtract NH4+ via ion exchange. Due to the multi-cationic population of its zeolites M was not effective enough in removing ammonium and, in the Minimum Inhibitory Concentration (MIC) test, allowed bacterial growth even at a concentration of 50 mg/mL. Inhibition was achieved with M-Na because it contained sodium zeolites capable of maximizing NH4+ subtraction, although the MIC was high (30 mg/mL). M-Zn evidenced a more effective inhibitory capacity, with a MIC of 4 mg/mL. Zinc has antimicrobial properties and H. pylori growth was affected by Zn2+ released from clinoptilolite and mordenite. These zeolites, being more selective towards NH4+ than Zn2+, can also subtract ammonium to the bacterium, thus enhancing the efficacy of M-Zn.

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