Bilayers merge even when exocytosis is transient

During exocytosis, the lumen of secretory vesicles connects with the extracellular space. In some vesicles, this connection closes again, causing the vesicle to be recaptured mostly intact. The degree to which the bilayers of such vesicles mix with the plasma membrane is unknown. Work supporting the kiss-and-run model of transient exocytosis implies that synaptic vesicles allow neither lipid nor protein to escape into the plasma membrane, suggesting that the two bilayers never merge. Here, we test whether neuroendocrine granules behave similarly. Using two-color evanescent field microscopy, we imaged the lipid probe FM4-64 and fluorescent proteins in single dense core granules. During exocytosis, granules lost FM4-64 into the plasma membrane in small fractions of a second. Although FM4-64 was lost, granules retained the membrane protein, GFP-phogrin. By using GFP-phogrin as a probe for resealing, it was found that even granules that reseal lose FM4-64. We conclude that the lipid bilayers of the granule and the plasma membrane become continuous even when exocytosis is transient.     

Status and perspectives of non invasive cell tracking

The interaction of different cells is an important regulator in the development of many diseases, including cancer. Some cells are recruited directly from the local tissue environment, others reach the pathological focus via the circulation. Using non-invasive cell tracking methods, the distribution and migration of labeled cells can be studied in experimental animal models, and the role of these cells on the pathogenesis of disease can thus be elucidated. Scintigraphy and SPECT, and especially MRI and optical imaging, are frequently used for this purpose. Studies are mostly performed with macrophages and granulocytes (inflammatory cells), which accumulate in nephritis, encephalitis, and tumors. At present, the understanding of progenitor cell migration and differentiation is gaining increasing interest in neurological disorders (for example Parkinson's disease) and in cardiac diseases (for example myocardial infarction). Non-invasive cell tracking is already established in basic research; in the future, a clinical application of cell tracking is foreseeable in the framework of cell therapy.     

Physical and biological characterization of superparamagnetic iron oxide- and ultrasmall superparamagnetic iron oxide-labeled cells: a comparison

RATIONALE: Superparamagnetic iron-oxide particles are used frequently for cellular magnetic resonance imaging and in vivo cell tracking. The purpose of this study was to compare the labeling characteristics and efficiency as well as toxicity of superparamagnetic iron oxide (SPIO) and ultrasmall superparamagnetic iron oxide (USPIO) for 3 cell lines. METHODS: Using human fibroblasts, immortalized rat progenitor cells and HEP-G2-hepatoma cells, dose- and time-dependence of SPIO and USPIO uptake were evaluated. The amount of intracellular (U)SPIO was monitored over 2 weeks after incubation by T2-magnetic resonance relaxometry, ICP-mass-spectrometry, and histology. Transmission-electronmicroscopy was used to specify the intracellular localization of the endocytosed iron particles. Cell death-rate and proliferation-index were assessed as indicators of cell-toxicity. RESULT: For all cell lines, SPIO showed better uptake than USPIO, which was highest in HEP-G2 cells (110 +/- 2 pg Fe/cell). Cellular iron concentrations in progenitor cells and fibroblasts were 13 +/- 1pg Fe/cell and 7.2 +/- 0.3pg Fe/cell, respectively. For all cell lines T2-relaxation times in cell pellets were below detection threshold (<3 milliseconds) after 5 hours of incubation with SPIO (3.0 micromol Fe/mL growth medium) and continued to be near the detection for the next 6 days. For both particle types and all cell lines cellular iron oxide contents decreased after recultivation and surprisingly were found lower than in unlabeled control cells after 15 days. Viability and proliferation of (U)SPIO-labeled and unlabeled cells were not significantly different. CONCLUSIONS: The hematopoetic progenitor, mesenchymal fibroblast and epithelial HEP-G2 cell lines accumulated SPIO more efficiently than USPIO indicating SPIO to be better suited for cell labeling. However, the results indicate that there may be an induction of forced cellular iron elimination after incubation with (U)SPIO.     

Microsurgical excision of symptomatic sacral perineurial cysts: a study of 15 cases

BACKGROUND: The widespread use of magnetic resonance imaging (MRI), now the first line investigation for back and leg pain, reveals cystic sacral lesions more often than myelography did in the past. There is agreement that symptomatic perineurial sacral cysts should be treated surgically. However, it is still debated whether the preference should be given to the curative option, consisting of excision of the cyst with duraplasty, or to drainage of the cyst to relieve symptoms. In this retrospective study the efficacy of microsurgical cyst resection with duraplasty is evaluated. METHODS: In 15 patients presenting with pain and neurologic deficits, myelography and/or MRI detected sacral cysts. The clinical features suggested that the space-occupying lesions caused the disturbances. Microsurgical excision of the cyst along with duraplasty or plication of the cyst wall was performed in all the cases. Postoperative care included bed rest and CSF drainage for several days. RESULTS: In 13 out of 15 patients the preoperative radicular pain disappeared after surgery. The 2 patients with motor deficits and the 6 patients with bladder dysfunction recovered completely. In all except 1 of the 10 patients complaining of sensory disturbances a significant improvement was achieved. No complications were observed. CONCLUSION: Microsurgical excision of the cyst combined with duraplasty or plication of the cyst wall is an effective and safe treatment of symptomatic sacral cysts and, in the view of the authors, the method of choice.     

Results of total aponeurectomy for Dupuytren’s contracture in 61 patients: a retrospective clinical study

Introduction  Many surgical techniques have been advocated to treat Dupuytren’s contracture. Partial fasciectomy is often performed to treat the whole spectrum of Dupuytren’s disease. Method  We have reviewed the effectiveness of total aponeurectomy performed on 61 patients [10 women and 51 men (male:female ratio 5.1:1) with a mean age of 63.0 (range 42–79 years) and a mean follow-up of 3.45 years (range 1.03–6.39 years)]. Results  Post-operative complications including haematoma, seroma or necrosis were found in 13.8% of the patients. Recurrence of contracture occurred in 10.8% of the patients and 4.6% of the operated patients presented with a nerve lesion. Nerve irritation occurred in 6.2% of the patients. The mean DASH-score was 3.85 (range 0–52.5). Family pre-disposition was an important risk factor for Dupuytren’s disease with 44.3% of patients having a positive family history. Conclusion  We suggest that total aponeurectomy is a promising alternative to partial fasciectomy with low risk for a recurrence and slightly increased risk for a nerve lesion.     

Retrospective motion gating in small animal CT of mice and rats

OBJECTIVES: Implementation and evaluation of retrospective respiratory and cardiac gating of mice and rats using a flat-panel volume-CT prototype (fpVCT). MATERIALS AND METHODS: Respiratory and cardiac gating was implemented by equipping a fpVCT with a small animal monitoring unit. ECG and breathing excursions were recorded and 2 binary gating signals derived. Mice and rats were scanned continuously over 80 seconds after administration of blood-pool contrast media. Projections were chosen to reconstruct volumes that fall within defined phases of the cardiac/respiratory cycle. RESULTS: Multireader analysis indicated that in gated still images motion artifacts were strongly reduced and diaphragm, tracheobronchial tract, heart, and vessels sharply delineated. From 4D series, functional data such as respiratory tidal volume and cardiac ejection fraction were calculated and matched well with values known from literature. DISCUSSION: Implementation of retrospective gating in fpVCT improves image quality and opens new perspectives for functional cardiac and lung imaging in small animals.     

Arthrodesis of the distal interphalangeal joint: description of a new technique and clinical follow-up at 2 years

INTRODUCTION: Arthrodesis of the distal interphalangeal joint (DIPJ) is an accepted operative procedure to treat osteoarthritis, instability and joint deformity. There is a wide spectrum of recommended operative techniques including cerclage wires, headless screws, bio-resorbable pins and lag screws. Lag screw fixation remains one of the most accepted methods of fixation; however, problems can occur in particular subsidence of the screw head leading to loss of compression and prominence of the screw head in the finger tip necessitating removal. We describe here a new technique of DIPJ arthrodesis that avoids these problems. METHODS AND PATIENTS: The technique is described here in detail, and clinical results are given for 18 joints (17 patients) followed-up at a mean 24 months (range 6-44). RESULTS: Mean post-operative DASH score was 15. There were no cases of non-union; only four complications were seen, one case of nail growth disturbance and one of early screw breakage necessitating revision. Two infections necessitated early hardware removal although both arthodesis were united. CONCLUSION: The technique is simple and reliable giving consistent clinical results.