Development of postbiotics by whey bioconversion with Enterococcus faecalis M157 KACC81148BP and Lactococcus lactis CAU2013 KACC81152BP for treating periodontal disease and improving gut health.

This study developed postbiotics with whey bioconversion product produced by Enterococcus faecalis M157 KACC 81148BP, and mixed whey bioconversion products produced by E. faecalis M157 KACC 81148BP and Lactococcus lactis ssp. lactis CAU2013 KACC 81152BP to alleviate periodontitis (PD) and to improve gut health. The powdered whey bioconversion product (EF) produced by E. faecalis M157 KACC 81148BP, mixed whey bioconversion products (EF+LL) from E. faecalis M157 KACC 81148BP and L. lactis CAU2013 KACC 81152BP, and phosphate-buffered saline (PBS; control) were administered orally to PD-induced rats for 8 wk. Infiltration of inflammatory cells and epithelial proliferation in periodontal tissue were found in control, but the lesions were reduced in PD+EF group (administration of EF to PD-induced rats), and no lesions were observed in PD+EF+LL group (administration of EF+LL to PD-induced rats). The bone loss volumes in PD+EF and PD+EF+LL groups were lower than in control. Cytokine production levels related to inflammation were lower and antioxidative stress markers were higher in PD+EF and PD+EF+LL groups than in control for both periodontal tissue and gut. The ratios of Lactobacillus spp. in gut microbiome of PD+EF and PD+EF+LL groups were higher than in control. These results indicate that the whey bioconversion product produced by E. faecalis M157 KACC 81148BP, and mixed whey bioconversion products produced by E. faecalis M157 KACC 81148BP and L. lactis CAU2013 KACC 81152BP are effective on relieving periodontitis and improving the gut health.

Fermented milk with Lactobacillus curvatus SMFM2016-NK alleviates periodontal and gut inflammation, and alters oral and gut microbiota.

This study aimed to analyze the effect of milk fermented with Lactobacillus curvatus SMFM2016-NK on periodontal diseases and gut health in a rat model. To improve the effect of Lb. curvatus SMFM2016-NK-fermented milk administration for relieving periodontitis, the periodontitis rat models were treated with the following for 4 wk: 10% skim milk (normal), periodontitis + 10% skim milk (negative control), periodontitis + Lactobacillus rhamnosus GG-fermented milk (positive control), and periodontitis + Lb. curvatus SMFM2016-NK-fermented milk (PD+LCFM). Transcriptional analysis of inflammatory cytokines [tumor necrosis factor α (TNF-α), IL-1ß, IL-6, and IL-10] was performed via quantitative reverse-transcription PCR. The changes in the oral and gut microbiomes after administering Lb. curvatus SMFM2016-NK-fermented milk were analyzed with metagenomics sequencing using DNA extracted from the oral gingival tissues and feces from the cecum of the rat models. After treatment with Lb. curvatus SMFM2016-NK-fermented milk, the relative gene expression levels of TNFA and IL1B in the gingiva decreased in the PD+LCFM group compared with those in the negative control group. In the oral microbiome, the proportion of the phylum Proteobacteria in the PD+LCFM group was lower than that in the negative control after treatment with Lb. curvatus SMFM2016-NK-fermented milk. For the effect in the gut, the relative gene expression levels of inflammatory cytokines in the colon between the normal and negative control groups were not different; however, the expression levels of TNFA and IL1B in the PD+LCFM and positive control groups, respectively, were lower than those in the negative control group. The composition and diversity of the gut microbiome differed among normal, periodontitis, and Lb. curvatus SMFM2016-NK-fermented milk treatment groups. These results indicate that Lb. curvatus SMFM2016-NK-fermented milk could alleviate periodontal and gut inflammation and change oral and gut microbiota.

Safe pseudoelastic limit range under torsional loading with Reciproc Blue.

AIM: To determine the distortional angle and torsional load at the pseudoelastic limit of Reciproc Blue instruments and to verify the safety of using pre-set dedicated motors designed for use with the original Reciproc instruments. METHODOLOGY: Two torsional conditions of Reciproc R25 and Reciproc Blue R25 were tested using a custom device. The first condition fixed the file tips at 3 mm and repetitively rotated them, with gradually increasing angles, from 10° to 270°, and the resulting torque was recorded (n = 15). The second test involved a single continuous rotation until fracture (n = 15). The pseudoelastic limits of the instruments were determined from their torque-rotation curves. For statistical analysis, two-way analysis of variance and t-tests were used, at a 95% significance level. Tested specimens were examined using field-emission scanning electron microscopy (FE-SEM). RESULTS: The angle at the pseudoelastic limit was significantly greater for Reciproc Blue than Reciproc, regardless of test mode (P < 0.05). When torsional loading was repeated, using gradually increasing rotational angles, the torsional resistance was significantly lower than for a single rotation (P < 0.05). However, under all test conditions, the pseudoelastic limit was below the pre-set 170° of the dedicated reciprocating motor. FE-SEM evaluation of the lateral aspects of the instruments revealed numerous longitudinal microcracks running along their long axis. For Reciproc Blue groups, the file shaft machining grooves were distorted after repetitive and continuous torsional tests. CONCLUSIONS: The 170° pre-set angle of the dedicated endodontic motors for the Reciproc system was safe for Reciproc Blue in single or time-restricted use.

Anatomical relationship of maxillary posterior teeth with the sinus floor and buccal cortex.

This study aimed to evaluate the possibility of root fenestration or oroantral communication by evaluating the distance from root apex to the sinus floor and buccal cortex in maxillary posterior teeth using cone-beam computed tomography (CBCT) images. The study included 2182 roots of the maxillary posterior teeth from 219 patients after reviewing CBCT images of 462 patients according to the location of roots by two endodontists. The distances from each root apex to the maxillary sinus floor and buccal and palatal cortices were evaluated according to sex and age, and the mean values were compared by one-way analysis of variance and Mann-Whitney U-test. The distance between root apex and maxillary sinus floor was the greatest in maxillary first premolars and shortest in the mesio-buccal roots of maxillary second molars. The distances from root apex to the buccal and palatal cortical bones were significantly greater in male patients than those in female patients (P < 0·05). The palatal roots of maxillary first molars exhibited the highest incidence as well as the greatest mean length (1·96 mm) of protrusion into the maxillary sinus. The distance from root apex to the sinus floor was found to increase with age, except in case of maxillary second premolars. Understanding the relationship of maxillary posterior teeth with the sinus floor and buccal cortex could provide clinicians valuable information to help reduce iatrogenic damage.

'Screw-in' tendency of rotary nickel-titanium files due to design geometry.

AIM: To examine the effect of several standard geometric characteristics of rotary instruments on the 'screw-in' forces and stresses generated on root dentine using 3D finite element analysis (FEA). METHODOLOGY: Four cross-sectional designs (triangular, slender-rectangular, rectangular and square) were evaluated. The area of the triangular cross-section and of the slender-rectangular model were the same. Another rectangular model had the same centre-core diameter as the triangular one. Each design was twisted into a file model with 5, 10 or 15 threads over its 16-mm-long working section. Three curved root canals were simulated as rigid surface models: θ = 15 degrees/R = 36 mm radius; θ = 30/R = 18; and θ = 45/R = 12. A commercial FEA package was used to simulate the file rotating in the canal to determine the 'screw-in' force and reaction torque on the instrument. RESULTS: Instruments of a square cross-section had the highest 'screw-in' force and reaction torsional stresses followed by the rectangle, the triangle design and the slender-rectangle design, respectively. The file with closer pitch generated lower stresses, compared with that with longer pitch. The greater the root canal curvature, the higher the 'screw-in' force and reaction torque generated. CONCLUSION: This study demonstrated that the 'screw-in' tendency depends on both the instrument geometry and canal curvature. Clinicians should be aware that certain instrument designs are prone to develop high 'screw-in' forces, requiring the operator to maintain control of the handpiece or to use a brushing action to prevent instruments being pulled into the canal.

The impact of the National Denture Service on oral health-related quality of life among poor elders.

The objective of this study was to assess the effects of the Korean National Denture Service (NDS) for poor elderly people requiring dentures on oral health-related quality of life (OHRQOL). Data from follow-up studies were collected from 439 subjects at eight public health centres who answered every question of a questionnaire, and the OHRQOL was measured at the baseline and at 3-month follow-up after receiving the NDS according to the type of denture provision. The multivariate linear mixed model with a public health centre as a random effect for the score change of Oral Health Impact Profile (OHIP)-14K was carried out to confirm the factors related to the improvement in OHRQOL. The mean OHIP-14K was 28.60 at the baseline time points, and there was a decrease in the OHIP-14 scores to 21.14 ± 12.52 at the 3-month follow-up of the removable partial denture beneficiaries. The changes in OHIP-14K among complete denture beneficiaries were 21.53 ± 12.01 for previously dentate subjects and 22.54 ± 11.12 for edentate subjects. The multivariate linear mixed model of dentate subjects demonstrated that the improvement in the OHRQOL was associated with the number of remaining teeth, satisfaction with denture and self-reported oral health status after 3 months. In the case of the edentate model, satisfaction with denture was the only factor related to the improvement in OHRQOL. This study revealed considerable improvement in OHRQOL among poor elderly people after NDS. Satisfaction with provision of dentures was associated with improvement in the OHRQOL.

Effect of germination temperature on characteristics of phytase production from barley.

The effects of germination temperature on the growth of barley seedlings for phytase production were studied at 15, 20 and 25 degrees C for 6-10 days. The growth rate of the barley seedlings was increased as the germination temperature was increased. The initial rate of total protein production was closely coupled to that of the barley growth, and the rate of total protein production tended to increase as the germination temperature was increased. SDS-PAGE analysis of total protein from the barley seedlings showed time-dependent appearance and disappearance of protein bands. Although no significant phytase activity was detected at zero time of germination, a significant increase in phytase activity up to 7.9-fold occurred during the first several days of germination then decreased. Phosphate production (viz. phytate degradation) in the barley seedlings occurred rapidly at the beginning of germination. However, the rate of production continued to decrease with further germination. A time lag of about 1-2 days between the rate of total protein production and that of phytase production was observed. Unlike the extent of total protein production, that of phytase production was similar irrespective of germination temperature. Partial purification of a crude enzyme extract by hydrophobic interaction chromatography resulted in two phytase fractions (PI and PII). Zymogram analysis demonstrated that PI had two bands with molecular masses of about 66 and 123 kDa while PII had one band corresponding to a molecular mass of about 96 kDa. The optimal temperature for PI was found to be 55 degrees C, while it was 50 degrees C for PII. The enzyme fraction PI had a pH optimum at 6.0, whereas the optimum pH for PII was found to be 5.0. Addition of 0.1% (v/v) Tween 80 was found to increase enzyme activity significantly (i.e., 167% for PI and 137% for PII). Phytate in cereals including barley, rice, corn and soybean degraded effectively by the treatment of the barley phytases.

Poly(ethylene oxide)-poly(propylene oxide)-poly(ethylene oxide)/poly(epsilon-caprolactone) (PCL) amphiphilic block copolymeric nanospheres. II. Thermo-responsive drug release behaviors.

Amphiphilic block copolymers composed of relatively hydrophilic PEO-PPO-PEO block copolymer (Pluronic) and poly (epsilon-caprolactone) with hydrophobic character were synthesized by ring-opening polymerization of epsilon-caprolactone in the presence of PEO-PPO-PEO block copolymer using stannous octoate as a catalyst. Pluronic/PCL block copolymeric nanospheres with core-shell structure were prepared by dialysis method. They showed the average diameter of 116-196 nm depending on the type of copolymer. All the nanosphere samples exhibited a narrow size distribution. The critical micelle concentrations of Pluronic/PCL amphiphilic block copolymers determined by fluorescence spectroscopy were lower than that of the common low molecular weight surfactant. Their core-shell structure was confirmed by 1H NMR spectroscopy. Pluronic/PCL block copolymeric nanospheres exhibited the reversible change of size depending on the temperature. Release behaviors of indomethacin from Pluronic/PCL block copolymeric nanospheres also showed temperature dependence and a sustained release pattern. In addition, cytotoxicity test using an MTT assay method revealed that these indomethacin-loaded Pluronic/PCL nanospheres could remarkably reduce the cell damage compared with the unloaded free indomethacin.

Poly(ethylene oxide)-poly(propylene oxide)-poly(ethylene oxide) (Pluronic)/poly(epsilon-caprolactone) (PCL) amphiphilic block copolymeric nanospheres. I. Preparation and characterization.

Amphiphilic block copolymers based on PEO-PPO-PEO block copolymer (Pluronic) and poly(epsilon-caprolactone) were synthesized by bulk polymerization. The structural analysis of Pluronic/PCL block copolymer was carried out using FT-IR, 1H NMR, GPC, WAXD, DSC and TGA measurements. To prepare copolymeric nanospheres with a micellar structure, Pluronic/PCL amphiphilic block copolymers were dialyzed against water. The size and size distribution of Pluronic/PCL block copolymeric nanospheres were examined by dynamic light scattering measurement. They showed an average diameter of 116 to 196 nm depending on the type of copolymer. All the nanosphere samples exhibited a narrow size distribution. The critical micelle concentrations of Pluronic/PCL amphiphilic block copolymers determined by fluorescence spectroscopy were lower than that of common low molecular weight surfactants. We confirmed the formation of stable copolymeric nanospheres through the solution behavior of amphiphilic block copolymer in selective solvents.

Clonazepam release from bioerodible hydrogels based on semi-interpenetrating polymer networks composed of poly(epsilon-caprolactone) and poly(ethylene glycol) macromer.

Poly(ethylene glycol)(PEG) macromers terminated with acrylate groups and semi-interpenetrating polymer networks (SIPNs) composed of poly(epsilon-caprolactone)(PCL) and PEG macromer were synthesized to obtain a bioerodible hydrogel. Polymerization of PEG macromer resulted in the formation of cross-linked gels due to the multifunctionality of macromer. Glass transition temperature (Tg) and melting temperature (Tm) of PEG networks and PCL in the SIPNs were inner-shifted, indicating an interpenetration of PCL and PEG chains. Water content in the SIPNs increased with increasing PEG weight fraction due to the hydrophilicity of PEG. The amount of clonazepam (CNZ) released from the SIPNs increased with higher content in the SIPNs, lower drug loading, lower concentration of PEG macromer during the SIPNs preparation, and higher molecular weight of PEG. In particular, a combination with low PEG content and low CNZ solubility in water led to long-term constant release from these matrices in vitro and in vivo.

Long term effects of temperature and substrate level on BNR with an external nitrification reactor.

This study was conducted with a BNR (biological nutrient removal) process with an external nitrification and switching arrangement of anoxic and oxic stages. It was observed that the SPRR (specific phosphorus release rate) and SDNR (specific denitrification rate) were greatly affected by the organic loads, and SDNR had a higher temperature effect than other kinetic rates including nitrification. It was further observed that the stoichiometric values like PHA (poly hydroxyalkanoate) stored for P release and PHA consumed for P uptake also varied. Variations of % Px (phosphorus content) and PHA as intracellular matter suggest the PAOs were more active at lower temperatures with this process configuration, where more than 70% of NH4-N and phosphorus were removed at temperatures below 10 degrees.

Coulombic forces in protein-RNA interactions: binding and cleavage by ribonuclease A and variants at Lys7, Arg10, and Lys66.

The interactions between bovine pancreatic ribonuclease A (RNase A) and its RNA substrate extend beyond the scissile P-O5' bond. Enzymic subsites interact with the bases and phosphoryl groups of the bound substrate. Those residues interacting with the phosphoryl group comprise the P0, P1, and P2 subsites, with the scissile bond residing in the P1 subsite. Here, the function of the P0 and P2 subsites of RNase A is characterized in detail. Lys66 (P0 subsite) and Lys7 and Arg10 (P2 subsite) were replaced with alanine residues. Wild-type RNase A and the K66A, K7A/R10A, and K7A/R10A/K66A variants were evaluated as catalysts for the cleavage of poly(cytidylic acid) [poly(C)] and for their abilities to bind to single-stranded DNA, a substrate analogue. The values of kcat and Km for poly(C) cleavage were affected by altering the P0 and P2 subsites. The kcat/Km values for poly(C) cleavage by the K66A, K7A/R10A, and K7A/R10A/K66A variants were 3-fold, 60-fold, and 300-fold lower, respectively, than that of wild-type RNase A. These values indicate that the P0 and P2 subsites contribute 0.70 and 2.46 kcal/mol, respectively, to transition-state binding. Binding experiments indicate that the P0 and P2 subsites contribute 0.92 and 1.21 kcal/mol, respectively, to ground-state binding. Thus, the P0 subsite makes a uniform contribution toward binding the ground state and the transition state, whereas the P2 subsite differentiates, binding more tightly to the transition state than to the ground state. In addition, nucleic acid binding to wild-type RNase A is strongly dependent on NaCl concentration, but this dependence is diminished upon alteration of the P0 or P2 subsite. The logarithm of Kd is a linear function of the logarithm of [Na+] over the range 0.018 M

The effects of sequential inoculation of mixed rumen protozoa on the degradation of orchard grass cell walls by anaerobic fungus Anaeromyces mucronatus 543.

The effects of protozoa on the degradation of plant cell walls (CW) during different growth stages of the fungus Anaeromyces mucronatus have been investigated. Since fungi show a marked lag in their in vitro cultures and many protozoa rapidly die during a prolonged incubation time, the effects of protozoa may vary according to the growth phase of the fungi. Therefore, the approach adopted was (i) to inoculate CW with fungus monoculture, (ii) to inoculate CW with fungus-protozoa coculture, or (iii) to sequentially inoculate fungal cultures that had been grown in CW for 24 (initial stage of growth), 48, and 72 h (late stage of growth) with mixed protozoa. When a fungus was associated with protozoa, a growth phase dependent effect was observed. Ruminal protozoa adversely affected the growth and activity when introduced in the initial growth stage of A. mucronatus, but a synergetic interaction was detected when added to late growth stage cultures. Although there is no immediate explanation for these results, the data suggested that protozoa can engulf the fungal zoospores, which are in ruminal fluids and (or) attached to small feed particles, but cannot engulf the fungal thallus that is tightly attached to feed particles by a rhizoidal system. Our data indicated that the protozoa did not influence cellulolysis by the fungi in exponential and (or) stationary phase, but they had a marked inhibitory effect on fungi that were in lag phase. Inhibition during lag phase could result from the protozoal predation of fungal zoospores that had failed to attach to substrates.

Adhesion promoters, their effect on the bond strength of metal brackets.

There are occasions where it is desired to increase the bond strengths of no-mix adhesives. Noncompliant patients and hypocalcified and fluorosed tooth enamel require higher bond strengths than usually obtained. It was the purpose of this investigation to study the effects of adhesion promoters to increase bond strengths. In vitro shear bond strengths that use the Instron machine (Instron Corp., Canton, Mass.) and thermocycling methods indicate the bond strengths of a no-mix adhesive bonding 80 gauge metal mesh brackets can be significantly improved through the following techniques: control (MPa 9.0), sandblasting (MPa 10.8), sandblasting and silanating (MPa 11.9), Rocatec system (MPa 10.8), Kulzer "silicoating" (MPa 13.2), and adhesion promoters--Megabond--(MPa 13.3). These adhesive promoters have been and are clinically being tested with promising results.

A planar amperometric creatinine biosensor employing an insoluble oxidizing agent for removing redox-active interferences.

A planar microchip-based creatinine biosensor employing an oxidizing layer (e.g., a PbO2 film), where interfering redox-active substances are broken (i.e., oxidized) to redox-inactive products, was developed to facilitate the microfabrication of the sensor and to provide improved, reliable determination of creatinine in physiological samples. The feasibility of using hydrophilic polyurethanes in permselective barrier membranes for creatinine biosensors and the effect of adding a silanizing agent (adhesion promoter) on the sensor performance (e.g., sensitivity, stability, and lifetime) are described. The proposed creatinine microsensor with a three-layer configuration, i.e., enzyme, protecting, and oxidizing layers, exhibits good electrochemical performance in terms of response time (t95% = 98 s at 100-->200 microM creatinine change), linearity (1-1000 microM, r = 0.9997), detection limit (0.8 microM), and lifetime (approximately 35 days). The creatinine biosensor devised in a differential sensing arrangement that compensates the erroneous results from creatine is considered to be suitable for assay of serum specimens.