Antibacterial adhesives are promising to inhibit biofilms and secondary caries. The objectives of this study were to synthesize and incorporate quaternary ammonium methacrylates into adhesives, and investigate the alkyl chain length effects on three-dimensional biofilms adherent on adhesives for the first time. Six quaternary ammonium methacrylates with chain lengths of 3, 6, 9, 12, 16 and 18 were synthesized and incorporated into Scotchbond Multi-Purpose. Streptococcus mutans bacteria were cultured on resin to form biofilms. Confocal laser scanning microscopy was used to measure biofilm thickness, live/dead volumes and live-bacteria percentage vs. distance from resin surface. Biofilm thickness was the greatest for Scotchbond control; it decreased with increasing chain length, reaching a minimum at chain length 16. Live-biofilm volume had a similar trend. Dead-biofilm volume increased with increasing chain length. The adhesive with chain length 9 had 37% live bacteria near resin surface, but close to 100% live bacteria in the biofilm top section. For chain length 16, there were nearly 0% live bacteria throughout the three-dimensional biofilm. In conclusion, strong antibacterial activity was achieved by adding quaternary ammonium into adhesive, with biofilm thickness and live-biofilm volume decreasing as chain length was increased from 3 to 16. Antibacterial adhesives typically only inhibited bacteria close to its surface; however, adhesive with chain length 16 had mostly dead bacteria in the entire three-dimensional biofilm. Antibacterial adhesive with chain length 16 is promising to inhibit biofilms at the margins and combat secondary caries.
Anti-Bacterial Agents ; pharmacology ; Biofilms ; Methacrylates ; pharmacology ; Microscopy, Confocal ; Quaternary Ammonium Compounds ; pharmacology

OBJECTIVETo prepare three quaternary ammonium salt (QAS) monomers, and to compare their antibacterial activities against four oral bacterial strains.

METHODSThree antibacterial monomers [methacryloxyethyl benzyl dimethyl ammonium chloride (DMAE-BC), methacryloxyethyl m-chloro benzyl dimethyl ammonium chloride (DMAE-m-CBC), methacryloxyethyl cetyl dimethyl ammonium chloride (DMAE-CB)] were synthesized according to the general structure of target monomers. Their antibacterial effects were investigated using the broth dilution test on Gram-positive and Gram-negative bacterial strains (Streptococcus mutans, Streptococcus sanguis, Porphyromonas gingivalis, Prevotella melaninogenica ).

RESULTSThree different monomers were successfully obtained. All the tested bacterial strains were susceptible to the three monomers, among which DMAE-CB exhibited the lowest minimal inhibitory concentrations (MIC) ranging from 1.2 to 4.8 mg/L.

CONCLUSIONSAll these three QAS monomers have different antibacterial activities against four oral bacteria strains. The data indicate that DMAE-CB may be a candidate antibacterial agent for oral infectious diseases.

Anti-Bacterial Agents ; chemistry ; pharmacology ; Dental Materials ; chemistry ; pharmacology ; Microbial Sensitivity Tests ; Quaternary Ammonium Compounds ; chemistry ; pharmacology

An ammonium-tolerant mutant of Actinobacillus succinogenes, YZ25, was obtained in the medium containing 61-242 mmol/L NH4+ after DES mutagenesis. Succinic acid produced by the mutant YZ25 reached 32.68 g/L when the medium contains 50 g/L glucose and 121 mmol/L ammonium, which was increased by 180.5% compared with that of the parent strain. The effects of different ammonium salts on the growth of the mutant and its metabolic response to high ammonium concentrations were investigated. The results showed that low ammonium concentration could improve the specific growth rates of the mutants, while high ammonium concentration inhibited cell growth. The ammonia-nitrogen half-inhibition constants (Ki) for different ammonium salts were as follows: 215 mmol/L for (NH4)2SO4, 265 mmol/L for NH4HCO3, 235 mmol/L for NH4Cl, and 210 mmol/L for NH4NO3. The process of ammonium inhibition on the mutant YZ25 was investigated in 3.0 L stirred fermenter. When NH4OH was used to buffer the pH, cell growth was not inhibited. However, production of succinic acid and consumption of glucose gradually decreased when cells entered the stationary phase, and the glucose could not be utilized completely at the end of fermentation. The possible ammonium inhibition mechanism was discussed based on the metabolic pathway of A. succinogenes.
Actinobacillus ; genetics ; growth & development ; metabolism ; Bioreactors ; Drug Tolerance ; Fermentation ; Industrial Microbiology ; Mutation ; Quaternary Ammonium Compounds ; metabolism ; pharmacology ; Succinic Acid ; metabolism

Deiters' cells are the supporting cells in organ of Corti and are suggested to play an important role in biochemical and mechanical modulation of outer hair cells. We successfully isolated functionally different K+ currents from Deiters' cells of guinea pig using whole cell patch clamp technique. With high K+ pipette solution, depolarizing step pulses activated strongly outward rectifying currents which were dose-dependently blocked by clofilium, a class III anti-arrhythmic K+ channel blocker. The remaining outward current was transient in time course whereas the clofilium-sensitive outward current showed slow inactivation and delayed rectification. Addition of 5 mM tetraethylammonium (TEA) further blocked the remaining current leaving a very fast inactivating transient outward current. Therefore, at least three different types of K+ current were identified in Deiters' cells, such as fast activating and fast inactivating current, fast activating slow inactivating current, and very fast inactivating transient outward current. Physiological role of them needs to be established.
Animals ; Ear, Inner ; Guinea Pigs* ; Guinea* ; Hair ; Hearing ; Organ of Corti ; Pharmacology ; Potassium Channels ; Potassium* ; Quaternary Ammonium Compounds ; Tetraethylammonium

OBJECTIVETo develop a resin composite incorporated with nano-antibacterial inorganic filler containing long-chain alkyl quaternary ammonium salt, and to measure its effect on human dental plaque microcosm biofilm.

METHODSA novel nano-antibacterial inorganic filler containing long-chain alkyl quaternary ammonium salt was synthesized according to methods introduced in previous research. Samples of the novel nano-antibacterial inorganic fillers were modified by a coupling agent and then added into resin composite at 0%, 5%, 10%, 15% or 20% mass fractions; 0% composite was used as control. A flexural test was used to measure resin composite mechanical properties. Results showed that a dental plaque microcosm biofilm model with human saliva as inoculum was formed. Colony-forming unit (CFU) counts, lactic acid production, and live/dead assay of biofilm on the resin composite were calculated to test the effect of the resin composite on human dental plaque microcosm biofilm.

RESULTSThe incorporation of nano-antibacterial inorganic fillers with as much as 15% concentration into the resin composite showed no adverse effect on the mechanical properties of the resin composite (P > 0.05). Resin composite containing 5% or more nano-antibacterial inorganic fillers significantly inhibited the metabolic activity of dental plaque microcosm biofilm, suggesting its strong antibacterial potency (P < 0.05).

CONCLUSIONThis novel resin composite exhibited a strong antibacterial property upon the addition of up to 5% nano-antibacterial inorganic fillers, thereby leading to effective caries inhibition in dental application.

Anti-Bacterial Agents ; pharmacology ; Biofilms ; drug effects ; Composite Resins ; chemistry ; Dental Caries ; prevention & control ; Dental Plaque ; Humans ; Lactic Acid ; Quaternary Ammonium Compounds ; pharmacology ; Saliva

The purpose of this study is to investigate the function of a novel potassium transporter gene (NrHAK1) isolated from Nicotiana rustica roots using yeast complement and real-time PCR technique. The complementary DNA (cDNA) of NrHAK1, 2 488 bp long, contains an open reading frame (ORF) of 2 334 bp encoding a protein of 777 amino acids (87.6 kDa) with 12 predicted transmembrane domains. The NrHAK1 protein shows a high sequence similarity to those of high-affinity potassium transporters in Mesembryanthemum, Phytolacca acinosa, Arabidopsis thaliana, and so on. We found that the NrHAK1 gene could complement the yeast-mutant defect in K+ uptake. Among several tissues surveyed, the expression level of NrHAK1 was most abundant in the root tip and was up-regulated when exposed to potassium starvation. Moreover, the transcript accumulation was significantly reduced by adding 5 mmol/L NH4+ to the solution. These results suggest that NrHAK1 plays an important role in potassium absorption in N. rustica.
Cation Transport Proteins ; chemistry ; genetics ; physiology ; Plant Proteins ; chemistry ; genetics ; physiology ; Potassium ; metabolism ; Quaternary Ammonium Compounds ; pharmacology ; Sodium ; pharmacology ; Tobacco ; genetics ; metabolism

OBJECTIVETo study the effects of carbon source and nitrogen source on callus growth, and flavonoid content in Glycyrrhiza uralensis.

METHODInduction and culture of callus were conducted in the media of different concentrations of sucrose, and fructose and ratio of NH4+/NO3- ,and flavonoid content was measured by HPLC.

RESULTThe results showed that fructose was superior to sucrose for callus growth and flavonoid formation, and the optimum concentration was 2%. The flavonoids content was 2 times higher than that of sucrose as carbon sources. In the range of 2% -6% of fructose concentration, the flavonoid content was decreased along with the concentration of fructose, but the licochalcone was increased 5-fold. The highest flavonoid content of 151.47 microg x g(-1) was obtained when the ratio of NH4+/NO3- in the medium was 1/2. NH; inhibited the callus growth and flavonoid formation of G. uralensis.

CONCLUSIONFructose as carbon source was superior to sucrose for callus growth and flavonoid formation, and NO3- was favorable to the callus growth and flavonoids accumulation.

Culture Media ; pharmacology ; Disaccharides ; pharmacology ; Flavonoids ; biosynthesis ; Fructose ; pharmacology ; Glycyrrhiza uralensis ; drug effects ; growth & development ; metabolism ; Nitrates ; pharmacology ; Nitrogen Compounds ; pharmacology ; Plants, Medicinal ; drug effects ; growth & development ; metabolism ; Quaternary Ammonium Compounds ; pharmacology ; Sucrose ; pharmacology ; Tissue Culture Techniques

OBJECTIVETo explore the effect and mechanism of iron on the vasodilating effect of interleukin-2 (IL-2) in the isolated aortic ring.

METHODSIsometric tension was recorded in response to drugs in organ bath. Ferric ammonium citrate (FAC) was added to the bath 30 min before phenylephrine (1 micromol/L), which was followed by IL-2 in a cumulative fashion. Spectrophotometry was used to determine the activity of nitric oxide synthase (NOS) of the thoracic aorta.

RESULTSFAC (0.1 - 10 micromol/L) alone did not affect the tension of rings,but inhibited the vasodilating effect of IL-2 (1 - 1,000 U/ml) in a dose dependent manner. IL-2(1, 10, 100, 1000 U/ml) decreased the aortic tension to (78.47+/-4.31)%, (66.86+/-5.55)%, (52.62+/-4.51)% and (42.39+/-4.27)% of pre-drug control, respectively. However, after incubation with 10 micromol/L FAC in the presence of IL-2, the aortic tension was reduced to (89.81+/-1.94)%, (86.13+/-3.11)%, (77.16+/-5.66)% and (68.76+/-5.69)% of pre-drug control, respectively. Pretreatment with L-arginine (1 mmol/L) abolished the inhibitory effect of FAC. Pretreatment with FAC attenuated the increased activity of NOS induced by IL-2 from (22.10+/-1.87)U/mg prot to (15.71+/-0.89)U/mg prot. High Ca(2+) (2.5 mmol/L) incubation did not change the inhibitory effect of FAC. Pretreatment with FAC attenuated the increased caffeine-releasable pool of Ca(2+) by IL-2. High K(+) (10 mmol/L) incubation abolished the inhibitory effect of FAC.

CONCLUSIONFAC inhibits the vasodilating effect of IL-2 in the isolated aortic ring,which may be mediated by decreasing the activity of NOS. Intracellular calcium release and inward rectifier potassium channel are involved in the inhibitory effect of FAC.

Animals ; Aorta, Thoracic ; drug effects ; physiology ; Arginine ; pharmacology ; Calcium ; metabolism ; Ferric Compounds ; pharmacology ; In Vitro Techniques ; Interleukin-2 ; pharmacology ; Male ; Nitric Oxide Synthase ; metabolism ; Potassium ; pharmacology ; Quaternary Ammonium Compounds ; pharmacology ; Rats ; Rats, Sprague-Dawley ; Vasodilator Agents ; pharmacology

OBJECTIVEThe effects of different nitrogen forms on the growth and photosynthetic characteristics of Prunella vulgaris were observed.

METHODWater culture experiments were carried out to observe the effects of NH4+: NO3- ratios(100:0, 75: 25, 50:50, 25:75, 0:100 in mmol x L(-1)) on the growth and photosynthetic characteristics of P. vulgaris.

RESULTThe leaf area, fresh biomass and P(n) of these cultivars increased with the increasing of NH4(+) -N: NO3(-) -N ratios, and they were found to be the highest in 25: 75 NH4(+) -N: NO3)-) -N. However, they decreased with the increasing NH4(+) -N: NO3(-) -N ratio further. P. vulgaris had a minimum leaf area, biomass, chlorophyll content and P(n) in pure ammonium group. The biggest chlorophyll and carotenoid contents were found in the ratios of NH4(+) -N to NO3(-) -N of 50: 50.

CONCLUSIONThe results indicated that properly increasing nitrate proportion could promote the growth and photosynthesis of P. vulgaris.

Biomass ; Chlorophyll ; Nitrogen ; chemistry ; metabolism ; pharmacology ; Photosynthesis ; drug effects ; Plant Leaves ; drug effects ; Prunella ; growth & development ; metabolism ; Quaternary Ammonium Compounds ; chemistry ; metabolism

Natural toxic substances have a bitter taste and their ingestion sends signals to the brain leading to aversive oral sensations. In the present study, we investigated chronological changes in c-Fos immunoreactivity in the nucleus tractus solitarius (NTS) to study the bitter taste reaction time of neurons in the NTS. Equal volumes (0.5 mL) of denatonium benzoate (DB), a bitter tastant, or its vehicle (distilled water) were administered to rats intragastrically. The rats were sacrificed at 0, 0.5, 1, 2, 4, 8, or 16 h after treatment. In the vehicle-treated group, the number of c-Fos-positive nuclei started to increase 0.5 h after treatment and peaked 2 h after gavage. In contrast, the number of c-Fos-positive nuclei in the DB-treated group significantly increased 1 h after gavage. Thereafter, the number of c-Fos immunoreactive nuclei decreased over time. The number of c-Fos immunoreactive nuclei in the NTS was also increased in a dose-dependent manner 1 h after gavage. Subdiaphragmatic vagotomy significantly decreased DB-induced neuronal activation in the NTS. These results suggest that intragastric DB increases neuronal c-Fos expression in the NTS 1 h after gavage and this effect is mediated by vagal afferent fibers.
Adjuvants, Immunologic/pharmacology ; Afferent Pathways/physiology ; Animals ; Injections/veterinary ; Ligands ; Male ; Proto-Oncogene Proteins c-fos/*metabolism ; Quaternary Ammonium Compounds/*pharmacology ; Rats ; Rats, Sprague-Dawley ; Receptors, G-Protein-Coupled/*metabolism ; Solitary Nucleus/*physiology ; Vagus Nerve/*drug effects/*physiology