Enzyme 1 of the phosphoenolpyruvate:sugar phosphotransferase system is involved in resistance to MreB disruption in wild-type and ∆envC cells.

Cell wall synthesis in bacteria is determined by two protein complexes: the elongasome and divisome. The elongasome is coordinated by the actin homolog MreB while the divisome is organized by the tubulin homolog FtsZ. While these two systems must coordinate with each other to ensure that elongation and division are coregulated, this cross talk has been understudied. Using the MreB depolymerizing agent, A22, we found that multiple gene deletions result in cells exhibiting increased sensitivity to MreB depolymerization. One of those genes encodes for EnvC, a part of the divisome that is responsible for splitting daughter cells after the completion of cytokinesis through the activation of specific amidases. Here we show this increased sensitivity to A22 works through two known amidase targets of EnvC: AmiA and AmiB. In addition, suppressor analysis revealed that mutations in enzyme 1 of the phosphoenolpyruvate:sugar phosphotransferase system (PTS) can suppress the effects of A22 in both wild-type and envC deletion cells. Together this work helps to link elongation, division, and metabolism.

Cell density-dependent antibiotic tolerance to inhibition of the elongation machinery requires fully functional PBP1B.

The peptidoglycan (PG) cell wall provides shape and structure to most bacteria. There are two systems to build PG in rod shaped organisms: the elongasome and divisome, which are made up of many proteins including the essential MreB and PBP2, or FtsZ and PBP3, respectively. The elongasome is responsible for PG insertion during cell elongation, while the divisome is responsible for septal PG insertion during division. We found that the main elongasome proteins, MreB and PBP2, can be inhibited without affecting growth rate in a quorum sensing-independent density-dependent manner. Before cells reach a particular cell density, inhibition of the elongasome results in different physiological responses, including intracellular vesicle formation and an increase in cell size. This inhibition of MreB or PBP2 can be compensated for by the presence of the class A penicillin binding protein, PBP1B. Furthermore, we found this density-dependent growth resistance to be specific for elongasome inhibition and was consistent across multiple Gram-negative rods, providing new areas of research into antibiotic treatment.

Bond strength of lithium disilicate to polyetheretherketone.

STATEMENT OF PROBLEM: Polyetheretherketone (PEEK) is a high-performance polymer that is increasingly used in dentistry, for example, as a framework for implant-supported fixed complete dentures. One protocol calls for individual lithium disilicate crowns to be cemented on preparation-shaped retentive elements on the framework. However, the flexibility and strength of the bonded system is unclear. PURPOSE: The purpose of this in vitro study was to compare the flexibility and strength of bonded lithium disilicate to PEEK with the bond between lithium disilicate and zirconia. MATERIAL AND METHODS: Fifteen PEEK (JUVORA Dental Disc), 15 zirconia (ArgenZ HT+), and 30 lithium disilicate (IPS e.max CAD) beam-shaped specimens (12.5×2×2 mm) were prepared. The ends of the PEEK beams were conditioned with 50-µm aluminum oxide airborne-particle abrasion, followed by primer (visio.link) and light-activated polymerization. Zirconia specimens were prepared with airborne-particle abrasion and primer (Monobond Plus). Lithium disilicate specimens were etched with 4.5% hydrofluoric acid (IPS Ceramic Etching Gel) and primed (Monobond Plus). The lithium disilicate specimens were cemented (Multilink Automix) to the PEEK and zirconia specimens. Light- and chemical-activated polymerization were used. Monolithic specimens of PEEK and zirconia (25×2×2 mm) were also prepared. All specimens were stored overnight in distilled water and submitted to a 4-point bend test in a universal testing machine at 0.5 mm/min crosshead speed until fracture, and the flexural modulus and strength were calculated. Differences among groups were statistically tested by using 1-way analysis of variance followed by the Student-Newman-Keuls post hoc test (α=.05). RESULTS: All bonded specimens fractured at their adhesive interface. Zirconia bonded to lithium disilicate specimens (29.7 ±8.8 MPa) were approximately 3 times stronger than PEEK bonded to lithium disilicate specimens (10.4 ±2.7 MPa) and approximately 12 times more rigid (78.5 ±6.7 GPa and 6.5 ±1.8 GPa, respectively). The flexure of monolithic PEEK was such that it did not fracture when loaded at 0.5 mm/min, while zirconia fractured at 413.9 ±38.5 MPa. Monolithic PEEK was approximately 37 times more flexible than monolithic zirconia (4.3 ±0.3 GPa and 157.2 ±7.2 GPa, respectively). All values were statistically significantly different except between the flexural moduli of monolithic PEEK and PEEK bonded to lithium disilicate. CONCLUSIONS: The bond strength between PEEK and lithium disilicate was significantly weaker than between zirconia and lithium disilicate. Monolithic zirconia was significantly stiffer than monolithic PEEK.

Metabolic testing does not alter distance running lower body sagittal kinematics.

BACKGROUND: Distance running fitness is commonly assessed using metabolic testing (MT). During MT, the runner must wear a mask that covers the nose and mouth. It is unclear if this increased challenge alters running kinematics and/or stride-to-stride variability (SSV). In this study we thoroughly assess the sagittal plane lower body joint angles. RESEARCH QUESTION: Are there significant differences between standard treadmill running kinematics and those collected during MT? METHODS: Twenty recreational runners participated (34.8 ± 10.0 years; 20+ miles per week). Six Vicon Bonita cameras were used to collect kinematic data (200 Hz). A metabolic cart (Parvo Medics TrueOne 2400) was used for heart rate (HR) collection and testing. Participants ran 4 × 4 min at preferred pace: 2 control runs (CON) and 2 MT runs. Ten strides were used to generate average stance and swing joint angle plots. The phase plots were compared for CON and MT and mean difference scores were calculated (to determine the kinematic change). SSV was determined by assessing the standard deviations among the 10 strides. Repeated measures ANOVA was used to test for significant differences among CON and MT trials. Reliability was assessed for 8 discrete joint angles using ICC analysis. RESULTS: There were no significant differences between CON and MT for both the joint angle plot comparisons and SSV. For the discrete kinematic measures, ICC scores were strong (0.89-0.99) between CON and MT. During MT, there were slight increases (p < 0.01) in HR (145 ± 14 vs. 147 ± 14) and RPE (10.4 ± 1.5 vs. 11.4 ± 1.5). SIGNIFICANCE: Results from this study support the validity of simultaneously conducting a kinematic and MT analysis. However, clinicians and performance coaches should be aware that 1) MT is slightly more physiologically demanding than CON and 2) approaches from this study can be used during individual assessments to confirm that kinematics are similar (between CON and MT).

Photophysics of untethered ZnTPP-fullerene complexes in solution.

The spectroscopy and dynamic behavior of the self-assembled, Soret-excited zinc tetraphenylporphyrin (ZnTPP) plus fullerene (C(60)) model system in solution has been examined using steady state fluorescence quenching, nanosecond time-correlated single photon counting, picosecond fluorescence upconversion, and picosecond transient absorption methods. Evidence of ground state complexation is presented. Steady-state quenching of the S(2) and S(1) fluorescence of ZnTPP by C(60) reveals that the quenching processes only occur in the excited complexes, are ultrafast, and proceed at different rates in the two states. Only uncomplexed ZnTPP is observed by fluorescence lifetime methods; the locally excited complexes are either dark or, more likely, rapidly relax to products that do not radiate strongly. Both short-range (Dexter) energy transfer and electron transfer relaxation mechanisms are evaluated. Picosecond transient absorption data obtained from the subtle differences between the spectra of Soret-excited ZnTPP with and without a large excess of added C(60) reveal the formation, on a subpicosecond time scale, of relatively long-lived charge-separated species. Soret excitation of ZnTPP···C(60) does not produce a quantitative yield of species in the lower S(1) excited state.