Measurement of apparent cell radii using a multiple wave vector diffusion experiment

It had been previously shown that an idealized version of the two‐wave‐vector extension of the NMR pulsed‐field‐gradient spin echo diffusion experiment can be used to determine the apparent radius of geometries with restricted diffusion. In the present work, the feasibility of the experiment was demonstrated in an NMR imaging experiment, in which the apparent radius of axons in white matter tissue was determined. Moreover, numerical simulations have been carried out to determine the reliability of the results. For small diffusion times, the radius is systematically underestimated. Larger gradient area, finite length gradient pulses, and a statistical distribution of radii within a voxel all have a minor influence on the estimated radius. Magn Reson Med, 2009. © 2009 Wiley‐Liss, Inc.     

Inducibility and potential role of MecA-gene-positive oxacillin-susceptible Staphylococcus aureus from colonized healthcare workers as a source for nosocomial infections

To determine the carrier rate of methicillin-susceptible mecA-positive Staphylococcus aureus (dormant MRSA) among healthcare workers (HCWs), 447 nurses and physicians from 13 general wards and intensive care units were investigated for nasal or oropharyngeal S. aureus carriage during one year whenever an MRSA patient was treated. Induction of phenotypic resistance in all mecA-positive oxacillin-susceptible aureus was attempted by 24 h exposure to oxacillin and cefotaxime. Organisms from the broth tube with the highest antibiotic concentration and visible growth after incubation were re-exposed for a total of seven repetitive exposures. Two mecA-negative oxacillin-susceptible S. aureus served as negative control. A population analysis before and after antibiotic exposure was performed. A third of the HCWs were found to be S. aureus carriers. Only three nurses were MRSA positive (0.7%). Seven isolates of dormant MRSA were isolated in six nurses and one doctor (1.6%). After four days of repetitive antibiotic exposure six of seven dormant MRSA were highly resistant to oxacillin. Resistance of the two control S. aureus without the mecA gene was not changed by repetitive antibiotic exposure. Two of the seven dormant MRSA were clonally related as shown by pulsed-field gel electrophoresis (PFGE). The PFGE pattern of one dormant MRSA (HCW) was identical to an MRSA (HCW). The pattern of another dormant MRSA was indistinguishable from an MRSA isolated from a patient who was treated at the same time on the same ward suggesting transmission from the HCW to the patient. Dormant MRSA may be isolated twice as often as MRSA from HCWs. Transmission to patients is possible, which may lead to clinical infections. It might be useful to screen methicillin-susceptible S. aureus isolates from HCWs for the mecA gene when recurrent infections with MRSA occur on a ward and a source cannot be found.     

Skin bacteria after chlorhexidine exposure—is there a difference in response to human β-Defensin-3?

We investigated whether exposure to sub-lethal concentrations of chlorhexidine digluconate (CHG) changed the response of five Staphylococcus spp. to human β-Defensin-3 (hBD-3). The change in response for each strain was determined in vitro with time–kill experiments in suspension by comparing the mean log₁₀ reduction caused by hBD-3 at 1.5 and 3 h in exposed and non-exposed bacteria. The identity of staphylococcal species was verified by DNA sequence homology in the gyrA genes in comparison with reference strains. Baseline sub-lethal concentrations allowing visible bacterial growth were between 0.0625 and 0.25 μg/ml. Sub-lethal CHG concentrations increased within 3 days in two isolates. For S. capitis 19/2, CHG-exposed cells were less susceptible to 0.5 μg/ml hBD-3 (log₁₀ reduction 0.78 versus 2.06 at 1.5 h; p < 0.001; t-test). For S. aureus, however, CHG-exposed cells were more susceptible to 1 μg/ml hBD-3. The observed changes between CHG-exposed and non-exposed cells did not indicate a general trend in response to hBD-3. Overall, we found no consistent evidence that 3 days of exposure to CHG changed the response of five Staphylococcus spp. to hBD-3. The use of CHG for skin antisepsis is, based on our data, unlikely to change the natural defence activity of hBD-3.     

Lymphatic Vessels in Pancreatic Islets Implanted Under the Renal Capsule of Rats

Transplantation of pancreatic islets necessitates an engraftment process, including revascularization of the graft. Studies of graft vasculature have demonstrated that islets become revascularized during the first post-transplant week through an angiogenic process. If this also involves lymphatic vessels is unknown. The aim of the present study was to functionally evaluate if lymphatic vessels, which are absent in endogenous islets, form after islet transplantation. To achieve this, inbred Wistar-Furth rats were transplanted with 250 syngeneic islets under the renal capsule. Intra-vital microscopy of the graft in combination with interstitial injection of Evans Blue was performed 1 week, 1 month or 9-12 months later. In all animals studied, there was drainage through intra-graft lymphatic capillaries emptying into larger lymphatic vessels associated with the renal capsule. The number was slightly lower 1 week post-transplantation. Most of the lymphatic capillaries were present in the graft stroma, rather than interspersed among the endocrine cells. In some animals, we were able to demonstrate dye in regional lymph nodes. We conclude that unlike endogenous islets, islet grafts develop a lymphatic drainage. Its functional importance and characteristics remain to be established. However, it can be speculated that immune reactions may be facilitated by the presence of lymphatic vessels.     

Efficacy of hand disinfectants against vancomycin-resistant enterococci in vitro.

Vancomycin-resistant enterococci (VRE) may be spread within a hospital via the contaminated hands of the healthcare worker. Effective hand disinfectants are necessary to break chains of transmission. We determined the bactericidal activity of 1-propanol, chlorhexidine digluconate (0.5 and 4%). Sterillium (45% 2-propanol, 30% 1-propanol and 0.2% mecetronium etilsulphate), Skinsept F (70% 2-propanol, 0.5% chlorhexidine digluconate and 0.45% hydrogen peroxide) and Hibisol (70% 2-propanol and 0.5% chlorhexidine gluconate) against 11 clonally distinct enterococcal isolates in a quantitative suspension test. Four isolates were vancomycin susceptible, four were vanA and the remainder vanB positive. Eight isolates were identified as Enterococcus faecium, two as Enterococcus faecalis and one as Enterococcus gallinarum. The investigator was blinded to the species and the genotype. Four parallel experiments were carried out for each isolate, each preparation, each dilution and each reaction time. 1-Propanol (60%), Sterillium, Skinsept F and Hibisol were all highly bactericidal after 15 and 30 s against VRE and vancomycin-susceptible enterococci (VSE) with reduction factors (RF) > 6.4, even in dilution of 50% (v/v). No significant difference was observed between vanA isolates, vanB isolates and VSE. Chlorhexidine digluconate (0.5% and 4%) was found to be less bactericidal after 30, 60 and 300 sec (RF < or = 2.5). The vanB genotype isolates were found to be significantly more susceptible to chlorhexidine (0.5%) than the vanA isolates (60 sec; one-way ANOVA model; P = 0.05). After 300 sec the vanB genotype isolates were found to be significantly more susceptible to chlorhexidine (0.5%) than the other two genotype isolates (P = 0.016). The vanA isolates were found to be significantly more susceptible to chlorhexidine (4%) than the vanB isolates (300 s; P = 0.024). E. faecium was found to be less susceptible to chlorhexidine than E. faecalis at all concentrations and reaction times, but significant differences between RF were only observed at 60 sec for both chlorhexidine concentrations (P < 0.05; t-test for independent samples). Propanol is much more effective against enterococci than chlorhexidine and combination of the two may be useful in providing an immediate and long lasting effect.     

SAR-reduced spin-echo-based Bloch-Siegert B(1)(+) mapping: BS-SE-BURST

Fast and accurate B(1)(+) mapping is possible using phase-based Bloch-Siegert (BS) methods. Importantly, the off-resonant pulses needed for BS B(1)(+) mapping methods can easily be implemented in multiple MR sequences. BS-based B(1)(+) mapping has thus been introduced for gradient echo (BS-FLASH), spin-echo (BS-SE), and Carr, Purcell, Meiboom, Gill (CPMG)-based multi-SE and turbo-SE sequences. When using SE and multi-SE/turbo-SE-based BS sequences, however, the high intrinsic specific absorption rates must be considered in clinical situations. This study introduces a fast BS B(1)(+) mapping method based on a SE-BURST sequence (BS-SE-BURST). With SE-BURST sequences, multiple low-magnitude excitation pulses are applied prior to the refocusing pulse. Thus, multiple and different phase-encoded echoes can be acquired per excitation cycle. Compared with a SE sequence, this excitation strategy results in a similar signal-to-noise ratio (SNR) per unit time but with reduced specific absorption rate. The proposed BS-SE-BURST sequence was implemented on a conventional 3 T whole body MRI scanner and applied successfully.