New Insights into Glycopeptide Antibiotic Binding to Cell Wall Precursors using SPR and NMR Spectroscopy

Glycopeptide antibiotics, such as vancomycin and teicoplanin, are used to treat life‐threatening infections caused by multidrug‐resistant Gram‐positive pathogens. They inhibit bacterial cell wall biosynthesis by binding to the D ‐Ala‐D ‐Ala C‐terminus of peptidoglycan precursors. Vancomycin‐resistant bacteria replace the dipeptide with the D ‐Ala‐D ‐Lac depsipeptide, thus reducing the binding affinity of the antibiotics with their molecular targets. Herein, studies of the interaction of teicoplanin, teicoplanin‐like A40926, and of their semisynthetic derivatives (mideplanin, MDL63,246, dalbavancin) with peptide analogues of cell‐wall precursors by NMR spectroscopy and surface plasmon resonance (SPR) are reported. NMR spectroscopy revealed the existence of two different complexes in solution, when the different glycopeptides interact with Ac2Kd AlaD AlaOH. Despite the NMR experimental conditions, which are different from those employed for the SPR measurements, the NMR spectroscopy results parallel those deduced in the chip with respect to the drastic binding difference existing between the D ‐Ala and the D ‐Lac terminating analogues, confirming that all these antibiotics share the same primary molecular mechanism of action and resistance. Kinetic analysis of the interaction between the glycopeptide antibiotics and immobilized AcKd AlaD AlaOH by SPR suggest a dimerization process that was not observed by NMR spectroscopy in DMSO solution. Moreover, in SPR, all glycopeptides with a hydrophobic acyl chain present stronger binding with a hydrophobic surface than vancomycin, indicating that additional interactions through the employed surface are involved. In conclusion, SPR provides a tool to differentiate between vancomycin and other glycopeptides, and the calculated binding affinities at the surface seem to be more relevant to in vitro antimicrobial activity than the estimations from NMR spectroscopy analysis.     

An epidemiological study of low back pain in professional drivers

The prevalence of low back pain (LBP) was investigated in 598 Italian professional drivers exposed to whole-body vibration (WBV) and ergonomic risk factors (drivers of earth moving machines, fork-lift truck drivers, truck drivers, bus drivers). The control group consisted of a small sample of 30 fire inspectors not exposed to WBV. Personal, occupational and health histories were collected by means of a structured questionnaire. Vibration measurements were performed on representative samples of the machines and vehicles used by the driver groups. From the vibration magnitudes and exposure durations, alternative measures of vibration dose were estimated for each subject. Daily vibration exposure, expressed in terms of 8-h energy-equivalent frequency-weighted acceleration, A(8), averaged 0.28-0.61 (range 0.10-1.18) m s⁻² rms in the driver groups. Duration of exposure to WBV ranged between 1 and 41 years. The 7-day and 12-month prevalence of LBP was greater in the driver groups than in the controls. In the professional drivers, the occurrence of 12-month LBP, high intensity of LBP (Von Korff pain scale score ?5), and LBP disability (Roland & Morris disability scale score ?12) significantly increased with increasing cumulative vibration exposure. Even though several alternative measures of vibration exposure were associated with LBP outcomes, nevertheless a more regular trend of association with LBP was found for vibration dose expressed as ∑a_vit_i (m s⁻² h), in which the frequency-weighted acceleration, a_v, and lifetime exposure duration, t, were given equal weight. In multivariate data analysis, individual characteristics (e.g. age, body mass index) and a physical load index (derived from combining manual materials handling and awkward postures) were significantly associated with LBP outcomes, while psychosocial work factors (e.g. job decision, job support) showed a marginal relation to LBP. This study tends to confirm that professional driving in industry is associated with an increased risk of work-related LBP. Exposure to WBV and physical loading factors at work are important components of the multifactorial origin of LBP in professional drivers.