Fluorescence immunohistochemistry and confocal scanning laser microscopy

We have developed a reliable technique for labeling and examining neural structures in soft tissues associated with articular joints and have tested it in human wrist joints under various specimen-related conditions. The labeling protocol employs an immunohistochemical process with a panneuronal marker (PGP 9.5) as the primary antibody and Alexa Fluor 488 as the fluorescing secondary antibody. Imaging was done using a confocal laser scanning microscope, which produced exceptionally detailed three-dimensional images of nerve endings and transiting nerve fibers from thick sections of wrist joint ligaments obtained from human cadavers. The protocol provided a practical postmortem window for specimen acquisition and processing without significant apparent worsening of image quality. The images produced are resistant to fading with repeated exposure to a fluorescent light source, which gives many opportunities for observation. Background staining is minimal, producing high contrast labeling of target tissues, which, in turn, enhances image analysis.     

Pneumococcaemia as a presenting sign in 3 cases of multiple myeloma

Multiple myeloma was diagnosed in 3 patients following episodes of pneumococcaemia associated with neutropenia and decreased serum concentration of normal immunoglobulins. Severe pneumococcal infection is commonly encountered during the course of multiple myeloma, but has not been stressed as a presenting feature of the disease.     

Cervical root stimulation in the diagnosis of radiculopathy

Cervical root stimulation (CRS) was compared with conventional EMG, nerve conduction, and late response studies in 34 patients with possible cervical radiculopathy. Cervical roots were stimulated by monopolar needles inserted into paraspinal muscles, recording compound muscle action potentials in biceps, triceps, and abductor digiti minimi muscles. In 18 patients with clinical evidence of radiculopathy, EMG was abnormal in 11 (61%), but CRS was abnormal in all 18. Of 16 patients with symptoms but no signs of radiculopathy, EMG was abnormal in 5 (31%) and CRS was abnormal in 9 (56%).     

Dopaminergic innervation of the parahippocampal and hippocampal regions in the rat

In spite of extensive investigations of the rat's meso-cortico-limbic system, the dopaminergic (DA) innervation of the hippocampal formation (HF) has not heretofore been visualized by histochemical techniques. However pharmacological and biochemical studies strongly suggested its existence. We used tyrosine-hydroxylase (TH) as an immunocytochemical marker of DA neurons in rats in which the noradrenaline cortical innervation was previously destroyed by neurotoxins. The absence of noradrenergic axons was routinely controlled with dopamine-beta-hydroxylase immunocytochemistry. TH-positive axons, thus DA axons, reach the HF primarily through the fimbria in which they occupy a specific lateral sector. They innervate the ventral and caudal parts of the HF, the subiculum and the adjacent CA1 hippocampal field being the main targets. This DA terminal field in the HF, matches with the area projecting toward the nucleus accumbens. Thus the hippocampo-striatal projections which represent a link of functional importance between the limbic and central motor systems might be modulated by the dopaminergic meso-cortico-limbic pathway. The present immunocytochemical study confirms the very dense innervation of the entorhinal cortex (EC): DA axons are organized in dense fiber islands and occupy primarily the superficial layers (I-II-III). Da innervation predominates in the lateral part of EC, which is the site of multimodal cortical afferences and therefore relays information between the whole neocortex and the hippocampus. Thus the DA system could play an important role of modulation on hippocampal and parahippocampal functions.     

Time-dependent changes of insufficiency fractures of the sacrum: intraosseous vacuum phenomenon as an early sign

Sacral insufficiency fractures develop over a period of time and show time-dependent changes. We report on 15 CT examinations of 5 patients with early-stage insufficiency fractures of the sacrum. In 4 patients only irregular sclerosis without distinct fracture lines was present in 7 of 8 fractures. Of these 4 patients; 3 exhibited intraosseous gas inclusions in a ventral part of a lateral mass; 5 of 8 fractures disclosed a ventral cortical break. When distinct fracture lines had developed in 1 patient, intraosseous vacuum phenomenon had disappeared. Fracture lines evolve over weeks to months and show central bone absorption. The fractures can heal as demonstrated in 4 of 6 fractures in 3 patients, can persist over 1 year without significant changes or can progress to pseudoarthrosis with bone destruction similar to neuropathic joint disease. Intraosseous vacuum phenomena can persist to this stage. Intraosseous vacuum phenomenon is recognized as a potential finding in the early stage of sacral insufficiency fracture, which also is true for irregular sclerosis and ventral cortical disruption. Correspondence to: A. Stäbler