The discovery and development of inhibitors of fatty acid amide hydrolase (FAAH)

A summary of the discovery and advancement of inhibitors of fatty acid amide hydrolase (FAAH) is presented.     

Atomic force microscopy and hydrodynamic characterization of the adhesion of <Emphasis Type="Italic">staphylococcus aureus</Emphasis> to hydrophilic and hydrophobic substrata at different pH values

Understanding the mechanism of the bacterial cell adhesion to solid surfaces is of great medical and industrial importance. Bacterial adhesion to inert surfaces, such as a catheter, and other indwelling devices can form biofilm, consequently cause severe morbidity and often fatal infections. Initial bacterial adhesion to the material surfaces is a complicated process that is affected by various physicochemical properties of both bacterial cells and substratum surfaces. The surface properties of the cells were characterized by the sessile drop technique. Moreover, the interfacial free energy of Staphylococcus aureus adhesion to the supporting materials was determined. The results showed that S. aureus examined at different pH levels could be considered hydrophilic. We noted hat the electron-donor character of S. aureus was important at intermediate pH (pH 5, pH 7, and pH 9) and it decreased at both limits acidic and basic conditions. In addition, the adhesion of Staphylococcus aureus ATCC 25923 to the hydrophilic glass and hydrophobic indium tin oxide (ITO)-coated glass surfaces at different pH values (2, 3, 5, 7, 9 and 11) was investigated using atomic force microscopy (AFM) and image analysis was assessed with the Mathlab^® program. The data analysis showed that cells (number of adhering cells to glass and ITO-coated glass surface) adhered strongly at acidic pH and weakly at alkaline pH. Also, S. aureus has the ability to attach to both hydrophobic and hydrophilic surfaces, but the adhesion was higher on hydrophobic surface.     

Appropriateness of empiric antimicrobial therapy with imipenem/colistin in severe septic patients: observational cohort study

Background

Empiric antimicrobial therapy (EAMT) using imipenem/colistin is commonly prescribed as a first line therapy in critically ill patients with severe sepsis. We aimed to assess the appropriateness of prescribing imipenem/colistin as EAMT in ICU patients.

Methods

A 3-year observational prospective study included ICU patients that required imipenem/colistin as EAMT. The EAMT was assessed according to microbiological and clinical outcomes. The outcomes were: delay in apyrexia, delay in the decrease of the biological inflammatory parameters (BIP), the requirement for vasoactive agents, bacteriological eradication, length of stay, ventilator days and 30-day mortality.

Results

79 administrations of EAMT in 70 patients were studied. EAMT was appropriate in 52% of the studied cases. An ICU stay?>?6 days was related to inappropriateness, and chronic respiratory failure was associated with appropriateness. In the appropriate EAMT group, we showed: earlier apyrexia, shorter delay in the decrease of the BIP and a reduced significant vasopressors requirement. Furthermore, EAMT improved survival with a median gain of 4 days. Inappropriate EAMT increased the mortality risk by six. The acquisition of NI in ICU was also an independent factor of mortality.

Conclusions

EAMT using imipenem-colistin was appropriate in half of the cases and inappropriateness was associated with an increased ICU mortality risk.

     

Physiological responses of small-sided vs. regular games in youth volleyball players

The aim of the present study was to evaluate the effects of two different formats of small-sided games (SSGs; 2 vs. 2 and 3 vs. 3) on physiological parameters in youth volleyball players. Twelve youth volleyball players (17.2 ± 7.44 years, 1.89 ± 0.6 m, 72.83 ± 8.57 kg) completed three different games formats (regular game (RG): 6 vs. 6, SSG3: 3 vs. 3 and SSG2: 2 vs. 2). associated with two pitch dimensions (i.e., 2 vs. 2 and 3 vs. 3 on 18 × 4.5 m; 6 vs. 6 on 18 × 9 m). Each player performed 4 × 5 min SSG with a recovery period of 1 min between bouts. All players were members of the same youth team and played in a professional league. They had at least 6 years of volleyball training and no current injuries. This study was carried out during the competitive period. Heart rate (HR), blood lactate ([La⁻]), and rating of perceived exertion (RPE) were measured. Compared to RG, physiological (i.e., HR and [La⁻]) and RPE responses were significantly higher during SSG3 and SSG2 (all p < 0.05; ƞ_p² = 0.77, ƞ_p² = 0.65, ƞ_p² = 0.30, respectively). Moreover, HR and RPE were significantly higher in SSG2 compared to SSG3. In contrast, no significant differences were observed in [La⁻] between SSG2 and SSG3. These results suggest that the number of players influences the exercise intensity in small-sided volleyball games in youth players. Therefore, coaches could benefit from incorporating SSGs to manipulate the exercise intensity in youth volleyball players.     

Fatty acid amide signaling molecules

Key studies leading to the discovery and definition of the role of endogenous fatty acid amide signaling molecules are summarized.     

An Improved Strategy for Easy Process Monitoring and Advanced Purification of Recombinant Proteins

In this work, a multifunctional expression cassette, termed Multitags, combining different and complementary functionalities, was designed and used to monitor the expression and the purification of two model proteins (Pfu DNA polymerase and Myosin-VIIa- and Rab-Interracting protein : MyRIP). Multitags contains two affinity purification tags, a polyhistidine sequence (10× His) and the streptavidin-binding peptide (SBP) and as a marker tag the heme-binding domain of rat cytochrome b5 followed by the TEV cleavage site. Using the Multitags as fusion partner, more than 90 % of both fusion proteins were produced in soluble form when expressed in Escherichia coli KRX. In addition, high purity (99 %) of recombinant proteins was achieved after two consecutive affinity purification steps. The expression cassette also demonstrated an accurate monitoring capability comparable to that of a dual recognition-based method. The choice of the SBP tag was considered as an integral process that included a method for tag removal. Thus, an immobilized TEV protease fixed on streptavidin–agarose matrix was used for the cleavage of fusion proteins. After digestion, both unprocessed fusion proteins and Multitags were retained on the proteolytic column via their SBP sequence, allowing cleavage and recovery of target proteins on one step. This combined approach may accelerate the development of optimized production processes, while insuring high product quality and a low production cost.