Sulfide silver amplification of ferritin iron cores in blood and bone marrow cells

Summary A short exposure of cell suspensions to gaseous hydrogen sulfide, appropriate fixations, and subsequent physical development of silver shells around sulfidated insoluble metals were used to amplify ferritin iron cores in blood and bone marrow cells. The methods described are suitable for both light microscopy and transmission electron microscopy. These techniques made it possible to visualize Prussian Blue stainable ferritin and haemosiderin, as well as a large variety of isoferritin iron and other smaller particles beyond the sensitivity of Prussian Blue staining. Admixtures of sulfidatible zinc and traces of other heavy metals had to be taken into consideration. For further research, adaptations of sulfide silver staining to microphysical analyses were developed. However, conventional energy dispersive X-ray analysis was not sensitive enough to signalize the presence of Fe in sulfide silver amplified iron cores of a single or a few ferritin molecule(s). Proton-induced X-ray emission was used to measure Fe and Zn down to 1 fg/single cell in unstained or sulfide silver stained smears on thin foils. However, multielement analysis of homogeneous cell concentrates was much easier to perform and far more sensitive. In advanced iron overload, highly increased sulfide silver staining was found in peripheral blood cells including lymphocytes, monocytes, eosinophils, basophils, and — in extreme cases — also in neutrophils and platelets.     

Effects of strontium ranelate administration on bisphosphonate-altered hydroxyapatite: Matrix incorporation of strontium is accompanied by changes in mineralization and microstructure

Strontium ranelate (SR) is one therapeutic option for reducing risk of fracture in osteoporosis. The effects of SR treatment on hydroxyapatite (HA) previously altered by bisphosphonate (BP) administration remain to be established. Patients who have received long-term BP treatment and present with persistent high fracture risk are of particular interest. Paired iliac crest biopsies from 15 patients post-BP therapy were subjected to a baseline biopsy and a follow-up biopsy after treatment with 2 g SR day^?1 after either 6 months (n = 5) or 12 months (n = 10). Dual energy X-ray absorptiometry scans, serum parameters and biochemical markers were obtained. Quantitative backscattered electron imaging and energy-dispersive X-ray analyses combined with micro-X-ray fluorescence determinations were performed to observe any mineralization changes. Static 2-D histomorphometry was carried out to evaluate cellular and structural indices. After 6 months of SR treatment, increases in osteoid surface and strontium content were observed, but no other indices showed significant change. After 12 months of SR treatment, there was a significant increase in bone volume and trabecular thickness, and further increases in strontium content and backscattered signal intensity. These structural changes were accompanied by increased numbers of osteoblasts and increased osteoid surface and volume. Additionally, low bone resorption, as measured by beta-cross-laps, and a low number of osteoclasts were observed. SR treatment led to increased strontium content within the BP–HA nanocomposites and to increased osteoid indices and bone volume, which is indicative of newly formed bone, while osteoclasts were still suppressed. These data points suggest that SR might be considered as a therapeutic option for patients following long-term BP treatment.     

Histologic reactions of the bone-implant zone and cortical bone area after long-term hip replacement]

29 femora with cemented hip endoprostheses and 17 age related controls were analyzed regarding different histological criteria. All specimens were processed to undecalcified ultra thin grindings and in addition a few to surface stained block-grindings. The reactions at the bone implant interface in cases without loosening of the implant are: accumulation of macrophages and multinucleated giant cells, fibrous tissue membranes with a mean thickness of 103 microns and mineralization defects near the cement. The mean rate of direct bone/bone-cement contact is 2.7% of the whole cement surface. The phenomenons at the interface were explained as being the result of micromovement and resulting from wear and tear. The cortical bone demonstrates a remarkable loss of bone (up to 60% after 12 years) following an increase of osteoclastic resorption with no change of osteoblast activity. The localization of the bone loss indicates a relation to the new load situation after implantation.     

Allocation of nonbirefringent wear debris: darkfield illumination associated with PIXE microanalysis reveals cobalt deposition in mineralized bone matrix adjacent to CoCr implants

Abrasive joint replacement material that accumulates in the tissue induces reciprocal effects between prosthesis material and organism. Since the limitations of brightfield and polarized light microscopy for foreign body analysis are well known, a method was applied that ensures the detailed histological assessment of nonbirefringent particles in periprosthetic soft and hard tissue. Cemented and cementless interface regions of five selected autopsy hip implant cases (2 x Endo-Modell Mark III, LINK, 1 x St. Georg Mark II, LINK, Germany; 2 x Spongiosa Metal II, ESKA, Germany) were viewed under darkfield illumination and subsequently analyzed with proton-induced X-ray emission (PIXE). Eight autopsy cases without implants served as controls. Using darkfield illumination technique, metallic particles became visible as luminous points under the microscope. The majority of particles in the samples from the cemented cases were degradation products of radiopaque bone cement. There was minimal evidence of metallic alloy particles in the soft tissues. However, a considerable quantity of heavy metal cobalt (Co) was found in the periprosthetic mineralized bone tissue, which was not observed in the controls. The periprosthetic concentration of cobalt ranged from 38 to 413 ppm. The findings demonstrate a correlation between cobalt concentration, time since implantation, and distance from the implant. Darkfield microscopy associated with PIXE enables a detailed histological assessment of metal particles in the tissue. In an effort to optimize biomechanics, implant design and implantation techniques, the contamination of soft and hard tissue with heavy metal degradation products deserves similar attention in terms of alloy assortment.     

Arztbriefe neurologischer Kliniken in der Sicht niedergelassener Neurologen und Nervenärzte

Eight discharge reports involving five diagnoses (anterior territory ischemic stroke, epilepsy, Parkinson's syndrome, multiple sclerosis, polyneuropathy) from five neurological departments were peer-reviewed by five neurologists working in out-patient (private) practice. The review considered the diagnosis, case history, clinical status, laboratory investigation, differential diagnosis and treatment. Criticism mainly involved the quality of the clinical assessment, lack of clinical status at discharge, narrow or incomplete differential diagnosis and the quality of the neurophysiological investigations for epilepsy and polyneuropathy. Improvement potential was seen for the speed of reporting, better comprehensibility, omission of irrelevant information, greater participation of experienced neurologists in report writing, and standardization.