Search

Introduction: This study aimed at assessing the quantitative effect of calcium hydroxide, 2% chlorhexidine gel, and 1.5% chlorhexidine linked to xanthan gel specifically against intratubular bacteria. Methods and Materials: Fifty-two semi-cylindrical bovine dentin specimens were infected with Enterococcus (E.) faecalis by centrifugation with subsequent 7-days incubation. The surface of specimens was disinfected with 3% H202. Scanning electron microscopy (SEM), confocal laser scanning microscopy (CLSM) and the count of bacterial colony-forming units (CFU/mg) were used to assess dentin infection. A total of 40 specimens were incubated for 2 weeks with one of the intracanal medication applied (10 samples for each group): 1) calcium hydroxide, 2) 2% chlorhexidine gel, 3) 1.5% chlorhexidine linked to xanthan gel and 4) sterile saline. Final passive ultrasonic irrigation with 3% sodium hypochlorite was performed in half of the total specimens. The effect of intracanal medications and irrigation against intratubular bacteria was assessed by bacterial culturing of dentin shavings. Two-Way ANOVA model was applied followed by post-hoc Tukey's test for multiple pair-wise comparisons of mean CFU/mg values. Results: SEM, CLSM, and bacterial culturing confirmed the absence of the surface biofilm on the root canal wall and showed vital intratubular bacteria at the depth up to 700 µm. Two-week application of 1.5% chlorhexidine with xanthan gel and 2% chlorhexidine gel significantly decreased intratubular bacterial counts compared with saline (P=0.0003 and P=0.0005, respectively). Subsequent passive ultrasonic irrigation with 3% sodium hypochlorite significantly reduced the number of intratubular bacteria in all groups except for the group with 1.5% chlorhexidine-xanthan gel (P=0.0054). Conclusion: This modified ex vivo model study showed ultrasonically activated irrigation with sodium hypochlorite had greater effect on intratubular bacteria counts compared with 2-week application of intracanal medications. [ABSTRACT FROM AUTHOR]

Metastable near‐beta Ti‐21.8Nb‐6Zr and Ti‐19.7Nb‐5.8Ta (at%) alloys were subjected to a thermomechanical treatment comprising cold rolling (CR) with a true strain of e = 0.3 and post‐deformation annealing (PDA) in the 500–900°C temperature range to ensure the superelastic behavior which is important for bone implants. It was found that PDA resulted in formation of about 1–2 μm‐thick oxide layer on the Ti‐Nb‐Zr and Ti‐Nb‐Ta alloy samples; the layer was mainly composed of TiO2, in rutile and anatase modifications. The structure, the phase and chemical compositions, and some surface‐sensitive properties of the alloys were compared to those of Ti‐50.7Ni and Ti‐Grade2 reference materials. These surface layers (especially that of the Ti‐Nb‐Zr alloy) demonstrated a promising combination of high cohesion strength (load causing surface layer fracture is over 25 N), hardness (∼12 GPa), and hydrophilicity (contact angle ∼40°). Surface modification by controlled oxidation during air annealing increases corrosion resistance and enhances in vivo osteoinductive properties of Ti‐Nb‐Zr alloys by changing the surface microrelief, increasing the surface wettability, and improving the mechanical characteristics, thus laying the foundation for the development of medical implants with prolonged service life. So, it was confirmed that the same thermomechanical treatment, which creates conditions for the superelastic behavior of the bulk metal (CR: e = 0.3 + PDA = 500–700°C for 1 hr), would also create a strong, protective and biocompatible layer on the implant surface. [ABSTRACT FROM AUTHOR]

Antimicrobial photodynamic inactivation is currently being widely considered as alternative to antibiotic chemotherapy of infective diseases, attracting much attention to design of novel effective photosensitizers. Carboranyl-chlorin-e6 (the conjugate of chlorin e6 with carborane), applied here for the first time for antimicrobial photodynamic inactivation, appeared to be much stronger than chlorin e6 against Gram-positive bacteria, such as Bacillus subtilis, Staphyllococcus aureus and Mycobacterium sp. Confocal fluorescence spectroscopy and membrane leakage experiments indicated that bacteria cell death upon photodynamic treatment with carboranyl-chlorin-e6 is caused by loss of cell membrane integrity. The enhanced photobactericidal activity was attributed to the increased accumulation of the conjugate by bacterial cells, as evaluated both by centrifugation and fluorescence correlation spectroscopy. Gram-negative bacteria were rather resistant to antimicrobial photodynamic inactivation mediated by carboranyl-chlorin-e6. Unlike chlorin e6, the conjugate showed higher (compared to the wild-type strain) dark toxicity with Escherichia coli ΔtolC mutant, deficient in TolC-requiring multidrug efflux transporters. [ABSTRACT FROM AUTHOR]

Extracts from mistletoe (Viscum album L.) contain three main toxic proteins the lectins MLI (also known as viscumin), MLII and MLIII. A catalytic subunit of the mistletoe plant toxic lectin MLIII has been cloned and expressed in Escherichia coli cells. The structure and immunochemical properties of recombinant MLIII A-subunit were investigated using a panel of monoclonal antibodies against ML-toxins. Ribosome-inactivating activity of the recombinant MLIII A-subunit was determined in a cell-free system exhibiting inhibition of endogenous protein synthesis. The comparative analysis of nucleotide and deduced amino acid sequences of the cloned MLIII A and the native MLI A-subunits was performed, re- vealing the main differences in the primary structure of these proteins. Antigenicity analysis of the MLIII A-subunit has revealed a new epitope D179E184 that is not present in viscumin. The role of toxic lectins with respect to the immunological properties of mistletoe extracts is discussed. [ABSTRACT FROM AUTHOR]

Pharmacological effects of mistletoe extracts are determined by the concentration of three toxic lectins: mistletoe lectin I (MLI, or viscumin), MLII, MLIII. These proteins, as well as ricin, belong to ribosome-inactivating proteins type 2 (RIP2). However, the extracts from the plant Ricinus communis, containing ricin, are highly toxic. Ricin is about 30 times more effective in cell culture than viscumin. The dissociation of subunits and the transmembrane transport of catalytic subunit into the cytoplasm are needed to obtain the cytotoxic effect of RIP2. In this paper, hybridomas producing monoclonal antibodies against catalytic subunits of ricin and viscumin are described. Monoclonal antibodies against different epitopes, including one localized in intra-subunit area of catalytic subunits of ricin and viscumin, do not inhibit the enzymatic activity of these proteins in cell-free system. These hybridomas are resistant to the cytotoxic action of native toxins. Protective effect of antibodies are about the same for both toxins, though the dissociation of the subunits of ricin is more effective. The causes of the differences in activity of plant toxins as pharmacological agents, and the importance of above mentioned epitopes for neutralizing antibodies at the clinical applications of mistletoe extracts are discussed. [ABSTRACT FROM AUTHOR]

New test systems which allow to detect with high sensitivity the presence of isolated subunits in mistletoe extracts subunits are proposed. Interaction of monoclonal antibodies MNA5 and mouse anti-MLA (mistletoe lectin I A-chain) immune serum with panel of synthetic octapeptides linked to the surface of polyethylene pins have been analyzed. Two main immunogenic epitopes in MLA, AETHL and DGVFNNP, were found. The second sequence can be part of the MNA5 antibody epitope as shown by antigenic prediction. Possible role of the isolated A-and B-chains of mistletoe lectins in pharmacological effects of plant extracts is discussed. [ABSTRACT FROM AUTHOR]

Introduction: This study aimed at assessing the quantitative effect of calcium hydroxide, 2% chlorhexidine gel, and 1.5% chlorhexidine linked to xanthan gel specifically against intratubular bacteria., Methods and Materials: Fifty-two semi-cylindrical bovine dentin specimens were infected with Enterococcus (E.) faecalis by centrifugation with subsequent 7-days incubation. The surface of specimens was disinfected with 3% H 2 O 2 . Scanning electron microscopy (SEM), confocal laser scanning microscopy (CLSM) and the count of bacterial colony-forming units (CFU/mg) were used to assess dentin infection. A total of 40 specimens were incubated for 2 weeks with one of the intracanal medication applied (10 samples for each group): 1) calcium hydroxide, 2) 2% chlorhexidine gel, 3) 1.5% chlorhexidine linked to xanthan gel and 4) sterile saline. Final passive ultrasonic irrigation with 3% sodium hypochlorite was performed in half of the total specimens. The effect of intracanal medications and irrigation against intratubular bacteria was assessed by bacterial culturing of dentin shavings. Two-Way ANOVA model was applied followed by post-hoc Tukey's test for multiple pair-wise comparisons of mean CFU/mg values., Results: SEM, CLSM, and bacterial culturing confirmed the absence of the surface biofilm on the root canal wall and showed vital intratubular bacteria at the depth up to 700  m. Two-week application of 1.5% chlorhexidine with xanthan gel and 2% chlorhexidine gel significantly decreased intratubular bacterial counts compared with saline ( P =0.0003 and P =0.0005, respectively). Subsequent passive ultrasonic irrigation with 3% sodium hypochlorite significantly reduced the number of intratubular bacteria in all groups except for the group with 1.5% chlorhexidine-xanthan gel ( P =0.0054)., Conclusion: This modified ex vivo model study showed ultrasonically activated irrigation with sodium hypochlorite had greater effect on intratubular bacteria counts compared with 2-week application of intracanal medications., Competing Interests: ‘None declared’.

Extracts from mistletoe (Viscum album L.) contain three main toxic proteins--the lectins MLI (also known as viscumin), MLII and MLIII. A catalytic subunit of the mistletoe plant toxic lectin MLIII has been cloned and expressed in Escherichia coli cells. The structure and immunochemical properties of recombinant MLIII A-subunit were investigated using a panel of monoclonal antibodies against ML-toxins. Ribosome-inactivating activity of the recombinant MLIII A-subunit was determined in a cell-free system exhibiting inhibition of endogenous protein synthesis. The comparative analysis of nucleotide and deduced amino acid sequences of the cloned MLIII A and the native MLI A-subunits was performed, revealing the main differences in the primary structure of these proteins. Antigenicity analysis of the MLIII A-subunit has revealed a new epitope D179-E184 that is not present in viscumin. The role of toxic lectins with respect to the immunological properties of mistletoe extracts is discussed.

New test systems which allow to detect with high sensitivity the presence of isolated subunits in mistletoe extracts subunits are proposed. Interaction of monoclonal antibodies MNA5 and mouse anti-MLA (mistletoe lectin I A-chain) immune serum with panel of synthetic octapeptides linked to the surface of polyethylene pins have been analyzed. Two main immunogenic epitopes in MLA, AETHL and DGVFNNP, were found. The second sequence can be part of the MNA5 antibody epitope as shown by antigenic prediction. Possible role of the isolated A- and B-chains of mistletoe lectins in pharmacological effects of plant extracts is discussed.

Pharmacological effects of mistletoe extracts are determined by the concentration of three toxic lectins: mistletoe lectin I (MLI, or viscumin), MLII, MLIII. These proteins, as well as ricin, belong to ribosome-inactivating proteins type 2 (RIP2). However, the extracts from the plant Ricinus communis, containing ricin, are highly toxic. Ricin is about 30 times more effective in cell culture than viscumin. The dissociation of subunits and the transmembrane transport of catalytic subunit into the cytoplasm are needed to obtain the cytotoxic effect of RIP2. In this paper, hybridomas producing monoclonal antibodies against catalytic subunits of ricin and viscumin are described. Monoclonal antibodies against different epitopes, including one localized in intra-subunit area of catalytic subunits of ricin and viscumin, do not inhibit the enzymatic activity of these proteins in cell-free system. These hybridomas are resistant to the cytotoxic action of native toxins. Protective effect of antibodies are about the same for both toxins, though the dissociation of the subunits of ricin is more effective. The causes of the differences in activity of plant toxins as pharmacological agents, and the importance of above mentioned epitopes for neutralizing antibodies at the clinical applications of mistletoe extracts are discussed.

The application of two immunization protocols and two screening systems has allowed to produce five hybridomas mlb5, mlb6, mlb7, mlb9 and Mbch1, secreting mAbs against different sites of viscumin B-subunit. On the base of mlb9 and Mbch1 hybridomas, the test-system has been developed, able to detect up to 5 ng/ml of viscumin and up to 1 ng/ml of its B-chain. Produced hybridomas and monoclonal antibodies will be used for the studies of intracellular transport of plant toxins. Monoclonal antibody mlb7 will be used for the studies of viscumin interactions with immune system of mammals.