Autolysin-mediated peptidoglycan hydrolysis is required for the surface display of Staphylococcus aureus cell wall-anchored proteins.

Peptidoglycan hydrolases, or autolysins, play a critical role in cell wall remodeling and degradation, facilitating bacterial growth, cell division, and cell separation. In Staphylococcus aureus, the so-called "major" autolysin, Atl, has long been associated with host adhesion; however, the molecular basis underlying this phenomenon remains understudied. To investigate, we used the type V glycopeptide antibiotic complestatin, which binds to peptidoglycan and blocks the activity of autolysins, as a chemical probe of autolysin function. We also generated a chromosomally encoded, catalytically inactive variant of the Atl enzyme. Autolysin-mediated peptidoglycan hydrolysis, in particular Atl-mediated daughter cell separation, was shown to be critical for maintaining optimal surface levels of S. aureus cell wall-anchored proteins, including the fibronectin-binding proteins (FnBPs) and protein A (Spa). As such, disrupting autolysin function reduced the affinity of S. aureus for host cell ligands, and negatively impacted early stages of bacterial colonization in a systemic model of S. aureus infection. Phenotypic studies revealed that Spa was sequestered at the septum of complestatin-treated cells, highlighting that autolysins are required to liberate Spa during cell division. In summary, we reveal the hydrolytic activities of autolysins are associated with the surface display of S. aureus cell wall-anchored proteins. We demonstrate that by blocking autolysin function, type V glycopeptide antibiotics are promising antivirulence agents for the development of strategies to control S. aureus infections.

Mechanochemical Cyclotrimerization: A Versatile Tool to Covalent Organic Frameworks with Tunable Stacking Mode.

We introduce the first mechanochemical cyclotrimerization of nitriles, a facile strategy for synthesizing triazine-containing molecules and materials, overcoming challenges related to carbonization and solubility. Conducting this solid-state approach in a mixer ball mill with 4-Methylbenzonitrile, we synthesize Tris(4-methylphenyl)-1,3,5-triazine quantitatively in as little as 90 minutes. Just as fast, this mechanochemical method facilitates the synthesis of the covalent triazine framework CTF-1 using 1,4 Dicyanobenzene. Material characterization confirms its porous (650 m2 g-1) and crystalline nature. Adjusting the induced mechanical energy allows control over the obtained stacking conformation of the resulting CTFs - from a staggered AB arrangement to an eclipsed AA stacking conformation. Finally, a substrate scope demonstrates the versatility of this approach, successfully yielding various CTFs.

Development and validation of a high-throughput whole cell assay to investigate Staphylococcus aureus adhesion to host ligands.

Staphylococcus aureus adhesion to the host's skin and mucosae enables asymptomatic colonization and the establishment of infection. This process is facilitated by cell wall-anchored adhesins that bind to host ligands. Therapeutics targeting this process could provide significant clinical benefits; however, the development of anti-adhesives requires an in-depth knowledge of adhesion-associated factors and an assay amenable to high-throughput applications. Here, we describe the development of a sensitive and robust whole cell assay to enable the large-scale profiling of S. aureus adhesion to host ligands. To validate the assay, and to gain insight into cellular factors contributing to adhesion, we profiled a sequence-defined S. aureus transposon mutant library, identifying mutants with attenuated adhesion to human-derived fibronectin, keratin, and fibrinogen. Our screening approach was validated by the identification of known adhesion-related proteins, such as the housekeeping sortase responsible for covalently linking adhesins to the cell wall. In addition, we also identified genetic loci that could represent undescribed anti-adhesive targets. To compare and contrast the genetic requirements of adhesion to each host ligand, we generated a S. aureus Genetic Adhesion Network, which identified a core gene set involved in adhesion to all three host ligands, and unique genetic signatures. In summary, this assay will enable high-throughput chemical screens to identify anti-adhesives and our findings provide insight into the target space of such an approach.

Staphylococcus aureus adhesion to the host.

Staphylococcus aureus is a pathobiont capable of colonizing and infecting most tissues within the human body, resulting in a multitude of different clinical outcomes. Adhesion of S. aureus to the host is crucial for both host colonization and the establishment of infections. Underlying the pathogen's success is a complex and diverse arsenal of adhesins. In this review, we discuss the different classes of adhesins, including a consideration of the various adhesion sites throughout the body and the clinical outcomes of each infection type. The development of therapeutics targeting the S. aureus host-pathogen interaction is a relatively understudied area. Due to the increasing global threat of antimicrobial resistance, it is crucial that innovative and alternative approaches are considered. Neutralizing virulence factors, through the development of antivirulence agents, could reduce bacterial pathogenicity and the ever-increasing burden of S. aureus infections. This review provides insight into potentially efficacious adhesion-associated targets for the development of novel decolonizing and antivirulence strategies.

Opening the pathway towards a scalable electrochemical semi-hydrogenation of alkynols via earth-abundant metal chalcogenides.

Electrosynthetic methods are crucial for a future sustainable transformation of the chemical industry. Being an integral part of many synthetic pathways, the electrification of hydrogenation reactions gained increasing interest in recent years. However, for the large-scale industrial application of electrochemical hydrogenations, low-resistance zero-gap electrolysers operating at high current densities and high substrate concentrations, ideally applying noble-metal-free catalyst systems, are required. Because of their conductivity, stability, and stoichiometric flexibility, transition metal sulfides of the pentlandite group have been thoroughly investigated as promising electrocatalysts for electrochemical applications but were not investigated for electrochemical hydrogenations of organic materials. An initial screening of a series of first row transition metal pentlandites revealed promising activity for the electrochemical hydrogenation of alkynols in water. The most active catalyst within the series was then incorporated into a zero-gap electrolyser enabling the hydrogenation of alkynols at current densities of up to 240 mA cm-2, Faraday efficiencies of up to 75%, and an alkene selectivity of up to 90%. In this scalable setup we demonstrate high stability of catalyst and electrode for at least 100 h. Altogether, we illustrate the successful integration of a sustainable catalyst into a scalable zero-gap electrolyser establishing electrosynthetic methods in an application-oriented manner.

Structure-Bioactivity Relationships of Lapatinib Derived Analogs against Schistosoma mansoni.

We recently reported a series of compounds for a solubility-driven optimization campaign of antitrypanosomal compounds. Extending a parasite-hopping approach to the series, a subset of compounds from this library has been cross-screened for activity against the metazoan flatworm parasite, Schistosoma mansoni. This study reports the identification and preliminary development of several potently bioactive compounds against adult schistosomes, one or more of which represent promising leads for further assessment and optimization.

Bacterial Anti-adhesives: Inhibition of Staphylococcus aureus Nasal Colonization.

Bacterial adhesion to the skin and mucosa is often a fundamental and early step in host colonization, the establishment of bacterial infections, and pathology. This process is facilitated by adhesins on the surface of the bacterial cell that recognize host cell molecules. Interfering with bacterial host cell adhesion, so-called anti-adhesive therapeutics, offers promise for the development of novel approaches to control bacterial infections. In this review, we focus on the discovery of anti-adhesives targeting the high priority pathogen Staphylococcus aureus. This organism remains a major clinical burden, and S. aureus nasal colonization is associated with poor clinical outcomes. We describe the molecular basis of nasal colonization and highlight potentially efficacious targets for the development of novel nasal decolonization strategies.

Distinct rapid and slow phases of salt-induced hypertension in Dahl salt-sensitive rats.

OBJECTIVE: To test the hypothesis that Dahl salt-sensitive (Dahl-S) rats exhibit distinct and separable phases of salt sensitivity. METHODS: Blood pressure (BP) telemetry was used to describe the detailed time course of salt-induced hypertension in Dahl-S rats and in hybrid rats derived from Dahl-S and Dahl salt-resistant strains. RESULTS: Switching to a high salt (4% NaCl) diet led to a biphasic increase in BP. Phase-1 reached a plateau in 4 days whereas phase-2 progressed slowly over the subsequent 5 weeks. In hybrid rats, phase-1 was present in each rat whereas phase-2 was absent in many individuals. A correlation of the amplitude of the first and second phases was of borderline significance in Dahl-S rats (P = 0.053, R2 = 0.44, n = 9) but was clearly significant in hybrid rats (P < 0.0001, R2 = 0.78, n = 22). Increases in BP were reversible following 1 week of high salt but progressively less so after 4 and 7 weeks. Estimation of the chronic pressure-natriuresis relationship suggests that phase-1 is attributable to a reduced slope of this relationship. In contrast, phase-2 corresponds with a further reduction in slope and a progressive and irreversible resetting of the relationship to higher BP levels. CONCLUSIONS: Two phases of salt sensitivity coexist and provide distinct contributions to salt-induced hypertension in Dahl-S rats. Our data also suggest that short-term measures of salt-sensitivity may be predictive of the effect of salt on the eventual progression of salt-induced hypertension.

Increased salt-sensitivity in endothelial nitric oxide synthase-knockout mice.

BACKGROUND: Although impaired nitric oxide production contributes importantly to salt-sensitivity, the role of the endothelial isoform of nitric oxide synthase (eNOS) has received little attention. In the present study we compared the effects of a high-salt diet on the blood pressure response of eNOS knockout (eNOS-/-) and control (eNOS+/+) mice. METHODS: Mean arterial pressure (MAP), heart rate, pulse pressure, and activity levels were recorded by telemetry in mice fed a regular-salt diet (0.7% NaCl) followed by 6 weeks on either a high-salt (8% NaCl) or regular-salt diet. RESULTS: The eNOS-/- mice exhibited a 15% increase in MAP and a 2- to 2.5-fold increase in salt-sensitivity relative to the control strain. Salt-induced increases in MAP were well sustained in eNOS-/-, whereas in eNOS+/+ the initial increase was biphasic. The effects of salt on MAP were particularly pronounced during locomotor activity, during the dark phase, and at the peak levels of MAP recorded over the course of the day. The high-salt diet also led to a transient increase in the proportion of time spent active. Levels of heart rate and pulse pressure were relatively unaffected by the high-salt diet. CONCLUSION: The eNOS-/- mice exhibit an increased blood pressure response to a high-salt diet. This finding suggests that eNOS normally provides an important contribution to the body's adaptation to a salt load and that reduced production of NO by eNOS may promote salt-sensitivity and salt-induced hypertension.

Flexibility is not Related to Stretch-Induced Deficits in Force or Power.

Previous studies have demonstrated that an acute bout of static stretching may cause significant performance impairments. However, there are no studies investigating the effect of prolonged stretch training on stretch-induced decrements. It was hypothesized that individuals exhibiting a greater range of motion (ROM) in the correlation study or those who attained a greater ROM with flexibility training would experience less stretch-induced deficits. A correlation study had 18 participants (25 ± 8.3 years, 1.68 ± 0.93 m, 73.5 ± 14.4 kg) stretch their quadriceps, hamstrings and plantar flexors three times each for 30 s with 30 s recovery. Subjects were tested pre- and post-stretch for ROM, knee extension maximum voluntary isometric contraction (MVIC) force and drop jump measures. A separate training study with 12 subjects (21.9 ± 2.1 years, 1.77 ± 0.11 m 79.8 ± 12.4 kg) involved a four-week, five-days per week, flexibility training programme that involved stretching of the quadriceps, hamstrings and plantar flexors. Pre- and post-training testing included ROM as well as knee extension and flexion MVIC, drop and countermovement jump measures conducted before and after an acute bout of stretching. An acute bout of stretching incurred significant impairments for knee extension (-6.1% to -8.2%; p < 0.05) and flexion (-6.6% to -10.7%; p < 0.05) MVIC, drop jump contact time (5.4% to 7.4%; p < 0.01) and countermovement jump height (-5.5% to -5.7%; p < 0.01). The correlation study showed no significant relationship between ROM and stretch-induced deficits. There was also no significant effect of flexibility training on the stretch-induced decrements. It is probable that because the stretches were held to the point of discomfort with all testing, the relative stress on the muscle was similar resulting in similar impairments irrespective of the ROM or tolerance to stretching of the muscle. Key PointsA correlation and training study were used to examine the effects of increased range of motion on stretch-induced changes in force and jump measuresAn acute bout of stretching incurred significant impairments for knee extension and flexion MVIC, drop jump contact time and countermovement jump height.Neither study showed any significant relationship between ROM and stretch-induced deficits.

First-year outcomes of beating heart coronary artery bypass grafting using proximal mechanical connectors.

BACKGROUND: To determine the extended results of mechanical connectors we compared the 1-year outcomes of patients having beating heart coronary artery bypass surgery with at least one sutured or mechanically connected proximal vein graft anastomosis. METHODS: From May 2001 to December 2001, 166 patients were identified as having undergone off-pump bypass grafting utilizing at least one St. Jude symmetry aortic connector (St Jude Medical Anastomotic Technology Group, St. Paul, MN). Follow-up for major adverse cardiac events (MACEs), which is defined as cardiac mortality, myocardial infarction, or revascularization of a previous target vessel, was obtained on 162 patients (97.6%). A control group of 159 patients was identified from a cohort of patients having beating heart surgery with one or more sutured proximal vein graft anastomosis in the preceding year. The MACE follow-ups were obtained in 136 patients (85.6%) by direct telephone contact. RESULTS: Patients with connectors showed an accelerated number of MACEs beginning approximately 180 days from the time of surgery and stabilizing at approximately 300 days. Logistic regression analysis identified the presence of diabetes as a significant preoperative risk factor predisposing patients to earlier onset of MACEs (p = 0.03) with an odds ratio of 2.9 (95% confidence interval, 1.1 to 7.6). Insulin dependent diabetics showed no differences between connector and control patients in the frequency or timing of MACEs. Connector patients using oral hypoglycemic agents demonstrated a significant deviation (p = 0.01) from a similar control population in the prevalence and timing of MACEs. CONCLUSIONS: Connector patients showed an increased incidence of early MACEs. These events were characterized by an increased requirement for early target vessel revascularization and were predominantly in noninsulin-dependent diabetics.

The role of thromboelastography in directing blood product usage in infant open heart surgery.

OBJECTIVE: : Thromboelastography (TEG) measures the dynamics of clot formation in whole blood and provides data that can guide specific blood component therapy. This study analyzed whether the implementation of TEG affected blood product utilization and overall hemostasis in infants (6 months and younger) undergoing open heart surgery. METHODS: : TEG values measured include R (time to fibrin formation), angle (fibrinogen formation), and MA (platelet function). Blood product usage, TEG values, and operative parameters were collected during surgery on 112 consecutive infants (66 acyanotic) undergoing open heart surgery within the first 6 months of life. Controls consisted of chart data on 70 consecutive patients (57 acyanotic) undergoing the same surgical procedures before implementation of TEG (pre-TEG). RESULTS: : Using TEG, the pattern of blood product utilization changed. Compared with the pre-TEG era, TEG era patients demonstrated a significant increase in fresh frozen plasma usage intraoperatively (4.74 vs. 1.83 mL/kg; P < 0.001) and reduced postoperative use of platelets (1.69 vs. 3.74 mL/kg; P = 0.006) and cryoprecipitate (0.89 vs. 1.95 mL/kg; P = 0.149). Chest tube drainage was significantly reduced at 1, 2, and 24 hours in the TEG group.TEG angle and MA measurements suggest that fibrinogen and platelets of cyanotic patients are more sensitive to hemodilution than the acyanotic patients. CONCLUSIONS: : TEG allows for proactive, goal-directed blood component therapy with improved postoperative hemostasis in infants undergoing cardiopulmonary bypass.

Phenotyping the level of blood pressure by telemetry in mice.

1. Using telemetry, arterial blood pressure (BP) can be measured directly over long periods in freely behaving animals without recent anaesthesia or surgery. In the present review, we discuss the strengths and limitations of this method and important considerations in using the method to characterize the BP level in mice. 2. A variety of informative statistics can be used to describe the BP level and we have made available a spreadsheet template for their calculation on a routine basis. The BP level is well summarized using the average value for an entire 24 h period or for the individual light and dark phases of the day. Such long-term averages exhibit less statistical variation than those of short recording periods. In addition, averages of the dark and light phases of the day convey information concerning circadian variations of BP. 3. The frequency distribution of BP samples provides additional information concerning the range of BP values recorded over the course of the day and can be described in terms of percentiles of the distribution that correspond with the minimum and maximum BP values and their span. 4. In mice, BP can be markedly affected by locomotor activity cycles that occur frequently throughout both the light and dark phases of the day. In addition, BP is strongly affected by ambient temperature and food intake, as well as potentially by other determinants of energy balance. Consideration of these factors may help improve accuracy and precision when phenotyping the BP level in mice.

Trunk muscle electromyographic activity with unstable and unilateral exercises.

The purpose of this cross-sectional study was to evaluate the effect of unstable and unilateral resistance exercises on trunk muscle activation. Eleven subjects (6 men and 5 women) between 20 and 45 years of age participated. Six trunk exercises, as well as unilateral and bilateral shoulder and chest presses against resistance, were performed on stable (bench) and unstable (Swiss ball) bases. Electromyographic activity of the upper lumbar, lumbosacral erector spinae, and lower-abdominal muscles were monitored. Instability generated greater activation of the lower-abdominal stabilizer musculature (27.9%) with the trunk exercises and all trunk stabilizers (37.7-54.3%) with the chest press. There was no effect of instability on the shoulder press. Unilateral shoulder press produced greater activation of the back stabilizers, and unilateral chest press resulted in higher activation of all trunk stabilizers, when compared with bilateral presses. Regardless of stability, the superman exercise was the most effective trunk-stabilizer exercise for back-stabilizer activation, whereas the side bridge was the optimal exercise for lower-abdominal muscle activation. Thus, the most effective means for trunk strengthening should involve back or abdominal exercises with unstable bases. Furthermore, trunk strengthening can also occur when performing resistance exercises for the limbs, if the exercises are performed unilaterally.

Modeling tetracycline antibiotic sorption to clays.

Sorption interactions of three high-use tetracycline antibiotics (oxytetracycline, chlortetracycline, tetracycline) with montmorillonite and kaolinite clays were investigated undervaried pH and ionic strength conditions. Sorption edges were best described with a model that included cation exchange plus surface complexation of zwitterion forms of these compounds. Zwitterion sorption was accompanied by proton uptake, was more favorable on acidic clay, and was relatively insensitive to ionic strength effects. Calcium salts promoted oxytetracycline sorption at alkaline pHs likely by a surface-bridging mechanism. Substituent effects among the compounds in the tetracycline class had only minor effects on sorption edges and isotherms under the same solution pH and ionic strength conditions. At low ionic strength, greater sorption to montmorillonite than kaolinite was observed at all pHs tested, even after normalizing for cation exchange capacity. These results indicate that soil and sediment sorption models for tetracyclines, and other pharmaceuticals with similar chemistry, must account for solution speciation and the presence of other competitor ions in soil or sediment pore waters.