A DnaA-dependent riboswitch for transcription attenuation of the operon.

Transcription attenuation in response to the availability of a specific amino acid is believed to be controlled by alternative configurations of RNA secondary structures that lead to the arrest of translation or the release of the arrested ribosome from the leader mRNA molecule. In this study, we first report a possible example of the DnaA-dependent riboswitch for transcription attenuation in . We show that (i) DnaA regulates the transcription of the structural genes but not that of the leader gene; (ii) DnaA might bind to rDnaA boxes present in the HisL-SL RNA, and subsequently attenuate the transcription of the operon; (iii) the HisL-SL RNA and rDnaA boxes are phylogenetically conserved and evolutionarily important; and (iv) the translating ribosome is required for deattenuation of the operon, whereas tRNA strengthens attenuation.

Expression activation of over 70% of genes during the first hour of infection.

The obligate intracellular bacterium has a unique developmental cycle that alternates between two contrasting cell types. With a hardy envelope and highly condensed genome, the small elementary body (EB) maintains limited metabolic activities yet survives in extracellular environments and is infectious. After entering host cells, EBs differentiate into larger and proliferating reticulate bodies (RBs).

Requirement of GrgA for infectious progeny production, optimal growth, and efficient plasmid maintenance.

Hallmarks of the developmental cycle of the obligate intracellular pathogenic bacterium are the primary differentiation of the infectious elementary body (EB) into the proliferative reticulate body (RB) and the secondary differentiation of RBs back into EBs. The mechanisms regulating these transitions remain unclear. In this report, we developed an effective novel strategy termed dependence on plasmid-mediated expression (DOPE) that allows for the knockdown of essential genes in .

Requirement of GrgA for infectious progeny production, optimal growth, and efficient plasmid maintenance.

, an obligate intracellular bacterial pathogen, has a unique developmental cycle involving the differentiation of invading elementary bodies (EBs) to noninfectious reticulate bodies (RBs), replication of RBs, and redifferentiation of RBs into progeny EBs. Progression of this cycle is regulated by three sigma factors, which direct the RNA polymerase to their respective target gene promoters. We hypothesized that the -specific transcriptional regulator GrgA, previously shown to activate σ66 and σ28, plays an essential role in chlamydial development and growth.

Robust Heat Shock Response in Lacking a Typical Heat Shock Sigma Factor.

Cells reprogram their transcriptome in response to stress, such as heat shock. In free-living bacteria, the transcriptomic reprogramming is mediated by increased DNA-binding activity of heat shock sigma factors and activation of genes normally repressed by heat-induced transcription factors. In this study, we performed transcriptomic analyses to investigate heat shock response in the obligate intracellular bacterium , whose genome encodes only three sigma factors and a single heat-induced transcription factor.

Eight-year Microtensile Bond Strength to Dentin and Interfacial Nanomechanical Properties of a One-step Adhesive.

Purpose: To evaluate the microtensile bond strength (µTBS) of a one-step self-etch adhesive (1-SEA) to dentin and its interfacial nanomechanical properties after 8 years of water storage.

Materials And Methods: Flat coronal dentin surfaces of extracted human third molars were bonded with a 1-SEA (Clearfil S3 Bond Plus, CS3+) and built up with a hybrid resin composite (Clearfil AP-X). After storage in water for 24 h or 8 years, non-trimmed stick-shaped specimens were fabricated from the central part of each bonded tooth and subjected to the µTBS test at a crosshead speed of 1.

Identification of a GrgA-Euo-HrcA Transcriptional Regulatory Network in Chlamydia.

Chlamydia trachomatis is an obligate intracellular bacterium whose unique developmental cycle consists of an infectious elementary body and a replicative reticulate body. Progression of this developmental cycle requires temporal control of the transcriptome. In addition to the three chlamydial sigma factors (σ, σ, and σ) that recognize promoter sequences of genes, chlamydial transcription factors are expected to play crucial roles in transcriptional regulation.

GrgA overexpression inhibits Chlamydia trachomatis growth through sigma- and sigma-dependent mechanisms.

The obligate intracellular bacterium Chlamydia trachomatis is an important human pathogen with a biphasic developmental cycle comprised of an infectious elementary body (EB) and a replicative reticulate body (RB). Whereas σ, the primary sigma factor, is necessary for transcription of most chlamydial genes throughout the developmental cycle, σ is required for expression of some late genes. We previously showed that the Chlamydia-specific transcription factor GrgA physically interacts with both of these sigma factors and activates transcription from σ- and σ-dependent promoters in vitro.

High-resolution mechanical mapping of the adhesive-dentin interface: The effect of co-monomers in 10-methacryloyloxydecyl dihydrogen phosphate.

The presence of 10-methacryloyloxydecyl dihydrogen phosphate (MDP) at the adhesive-dentin interface enables ionic binding to calcium salts, which results in rigid nano-layering within the submicron scale resin-dentin interdiffusion zone. MDP has been used with additional co-monomers, such as hydroxyethyl methacrylate (HEMA) and/or 4-methacryloyloxyethyl-trimellitic acid (4-MET), mainly to enhance the chemical bonding properties. However, the use of co-monomers may compromise the rigidity of the adhesive-dentin interface.

Nonspecific toxicities of Streptococcus pyogenes and Staphylococcus aureus dCas9 in Chlamydia trachomatis.

Chlamydiae are common, important pathogens for humans and animals alike. Despite recent advancement in genetics, scientists are still searching for efficient tools to knock out or knock down the expression of chromosomal genes. We attempted to adopt a dCas9-based CRISPR interference (CRISPRi) technology to conditionally knock down gene expression in Chlamydia trachomatis using an anhydrotetracycline (ATC)-inducible expression system.

Quantitative/qualitative analysis of adhesive-dentin interface in the presence of 10-methacryloyloxydecyl dihydrogen phosphate.

Dental adhesive provides effective retention of filling materials via adhesive-dentin hybridization. The use of co-monomers, such as 10-methacryloyloxydecyl dihydrogen phosphate (10-MDP), is thought to be crucial for hybridization owing to their ionic-binding to calcium and co-polymerization in the polymerizable adhesives. Optimal hybridization partly depends on the mechanical properties of polymerized adhesives, which are likely to be proportional to the degree of conversion ratio.

Exceptional contact elasticity of human enamel in nanoindentation test.

Objective: Tooth enamel has unsurpassed hardness and stiffness among mammalian tissue structures. Such stiff materials are usually brittle, yet mature enamel can survive for a lifetime. Understanding the nanoscale origin of enamel durability is important for developing advanced load-bearing biomaterials.

Nanomechanical characterization of time-dependent deformation/recovery on human dentin caused by radiation-induced glycation.

An increase in non-enzymatic collagen matrix cross-links, such as advanced glycation end-products (AGEs), is known to be a major complication in human mineralized tissues, often causing abnormal fractures. However, degradation of mechanical properties in relation to AGEs has not been fully elucidated at the material level. Here, we report nanoscale time-dependent deformation and dimensional recovery of human tooth dentin that has undergone glycation induced by x-ray irradiation.

DnaA and LexA Proteins Regulate Transcription of the Gene in : The Role of DnaA in the Control of the SOS Regulon.

The gene belongs to the SOS network, encoding a key component of the nucleotide excision repair. The promoter region contains three identified promoters with four LexA binding sites, one consensus and six potential DnaA binding sites. A more than threefold increase in transcription of the chromosomal gene is observed in both the Δ Δ cells and cells, and a fivefold increase in the Δ Δ cells relative to the wild-type cells.

Effect of sodium alginate on mouse ovary vitrification.

The survival rate of vitrified-thawed ovarian tissues after autotransplantation still needs to be improved. Therefore finding an ideal cryoprectant to reduce the damage to ovaries that caused by vitrification will pave the way for application of ovary cryopreservation on clinics. Experiments were conducted to investigate the effect of sodium alginate in cryoprotectant solution on mouse ovaries during the vitrification process.

The advantages of endophyte-infected over uninfected tall fescue in the growth and pathogen resistance are counteracted by elevated CO.

Atmospheric CO concentrations are predicted to double within the next century. Despite this trend, the extent and mechanisms through which elevated CO affects grass-endophyte symbionts remain uncertain. In the present study, the growth, chemical composition and pathogen resistance of endophyte-infected (E+) and uninfected (E-) tall fescue were compared under elevated CO conditions.

Trans-translation ensures timely initiation of DNA replication and DnaA synthesis in Escherichia coli.

The trans-translation pathway, mediated by the transfer messenger RNA (tmRNA; encoded by the ssrA gene) and the SmpB protein (tmRNA-binding protein expressed in Salmonella enterica), which is conserved in bacteria, is required for various cellular processes. A previous study has shown that trans-translation is required to ensure timely (non-delayed) dnaA transcription and consequent initiation of DNA replication in Caulobacter crescentus. In this study, we observed that initiation of chromosome replication was delayed in Escherichia coli lacking the smpB and/or ssrA genes (DssrA, DsmpB, or DsmpBDssrA mutants).

Enhanced in vitro biological activity generated by surface characteristics of anodically oxidized titanium--the contribution of the oxidation effect.

Anodically oxidized titanium surfaces, prepared by spark discharge, have micro-submicron surface topography and nano-scale surface chemistry, such as hydrophilic functional groups or hydroxyl radicals in parallel. The complexity of the surface characteristics makes it difficult to draw a clear conclusion as to which surface characteristic, of anodically oxidized titanium, is critical in each biological event. This study examined the in vitro biological changes, induced by various surface characteristics of anodically oxidized titanium with, or without, release of hydroxyl radicals onto the surface.

Strain-rate stiffening of cortical bone: observations and implications from nanoindentation experiments.

While bone mineralization is considered to be responsible for its stiffness, bone durability partially associated with the time-dependent viscoelasticity of matrix proteins is still poorly elucidated. Here we demonstrate a novel mechanism of highly mineralized bone durability almost independent of inherent viscoelastic behaviour along with a protocol for measuring the mechanical properties of mineralized tissues. Strain-rate nanoindentation tests showed substantial stiffening of the highly mineralized calvarial bone, whereas large creep or stress relaxation was observed during constant load or displacement tests, respectively.

Nanomechanical properties and molecular structures of in vitro mineralized tissues on anodically-oxidized titanium surfaces.

Unlabelled: The biomechanical stability of mineralized tissues at the interface between implant surface and bone tissue is of critical importance. Anodically oxidized titanium prepared in a chloride solution results in enhanced mineralization of adherent osteoblasts and has antimicrobial activity against oral microorganisms. We evaluated the nanomechanical properties and molecular structures of the in vitro mineralized tissues developing around anodically oxidized titanium surfaces with and without preparation in chloride solution.

Lysyl oxidase like-2 reinforces unsatisfactory ossification induced by bone morphogenetic protein-2: relating nanomechanical properties and molecular changes.

Unlabelled: Bone morphogenetic protein-2 (BMP2) is among the most popular anabolic agents and substantially increase bone volume related to enhanced osteoblast differentiation. Here we demonstrate a remarkable deterioration in the nanomechanical properties of mineralized tissue induced from osteoblasts solely by the function of BMP2. Mineralized tissue of primary osteoblasts cultured with BMP2 shows molecular features of both bone and cartilage, but depletion of lysyl oxidase family members leads to poor nanomechanical properties of the mineralized tissue.

Antioxidant and osteogenic properties of anodically oxidized titanium.

Cells adhering onto implant surfaces are subjected to oxidative stress during wound healing processes. Although titanium and its alloys are among the most frequently used biomaterials in orthopedic and dental implants, titanium surfaces do not have antioxidant properties, and cells grown on these surfaces can show permanent oxidative stress. The present study assessed the antioxidant property and osteogenic properties of titanium samples with or without oxidation treatments.