Rhythmical slow wave electroencephalographic activity elicited by hippocampal injection of muscarinic agents in the rat

In freely moving male Wistar rats the hippocampal EEG was recorded by using chemitrodes (combinations of guide cannulas with bipolar recording electrodes) chronically implanted into the dorsal hippocampus (CA1).The muscarinic agents oxotremorine and arecoline applied intrahippocampally caused a long-lasting increase in the amount of rhythmical slow wave activity (RSA) of the hippocampal EEG accompanied by a decrease of the dominating frequency. At the same time, the well-known relationship between defined behavioral states and EEG patterns was abolished. The effect of oxotremorine could be blocked by subsequent application of the muscarinic antagonist scopolamine.The results support the hypothesis that a non-rhythmic activation of hippocampal muscarinic receptors may result in the generation of RSA within the hippocampal formation.     

Tacrolimus ointment causes inflammatory dendritic epidermal cell depletion but no Langerhans cell apoptosis in patients with atopic dermatitis

BACKGROUND: The topical immunomodulators tacrolimus and pimecrolimus are novel therapeutic options for atopic dermatitis (AD). The inhibition of nuclear factor of activated T cell-dependent proinflammatory cytokine production in cutaneous lymphocytes is an established effect of topical immunomodulators, which additionally influence mast cells, eosinophils, and dendritic cells (DCs). The latter include a reduced expression of the high-affinity IgE receptor FcepsilonRI, a reduced stimulatory capacity of lesional DCs, and a selective depletion of the inflammatory dendritic epidermal cells (IDECs) but not of Langerhans cells (LCs) from the lesional skin. OBJECTIVE: Because induction of apoptosis in lymphocytes is a reported tacrolimus effect, we asked whether tacrolimus ointment induces apoptosis of LCs or IDECs in AD lesions. METHODS: Epidermal single-cell suspensions were prepared from AD lesions of 9 tacrolimus-treated and 5 hydrocortisone butyrate-treated patients with AD before and after 1 week of treatment. Cell numbers, apoptosis rate, and immunophenotype were assessed by using the standardized FACS technique with terminal deoxynucleotidyltransferase-mediated dUTP nick end labeling, Annexin V, and 3-color immunophenotyping. Freshly isolated LCs and monocyte-derived DCs served as in vitro controls. RESULTS: Tacrolimus and steroid ointment induced a selective depletion of IDECs from the epidermis and reduced the expression of the costimulatory molecules CD80 and CD86. Tacrolimus ointment did not increase the rate of apoptotic DCs, whereas steroid ointment did so. The isolation-induced high apoptosis rate of freshly isolated LCs was unaffected by both drugs. CONCLUSION: Tacrolimus ointment selectively depletes IDECs and alters the immunophenotype of epidermal DCs in AD lesions, but there is no evidence for tacrolimus-induced DC apoptosis in this phenomenon.     

Localization of neuroglobin protein in the mouse brain

Neuroglobin is a recently discovered vertebrate oxygen-binding respiratory protein. In situ hybridization data demonstrated that neuroglobin-mRNA is widely expressed in neuronal cells of the central and peripheral nervous systems as well as in endocrine cells. The present study was conducted to investigate the presence of neuroglobin protein in neurons of the mouse brain. A polyclonal antibody directed against a synthetic peptide of neuroglobin was raised in rabbits and affinity-purified. The specificity of the antibody was demonstrated by ELISA and preabsorption tests. We report here for the first time that neuroglobin is expressed on the protein level in many brain sites including cerebral cortical regions, subcortical structures such as thalamus and hypothalamus, nuclei of cranial nerves in the brainstem and cerebellum. Thus, the widespread distribution of neuroglobin protein is in good agreement with its mRNA localization. Regionally differing intensities of immunostaining suggest different levels of neuroglobin protein expression, in line with the idea that brain regions show variation in their tolerance towards hypoxic conditions.     

Conservation of L-type Ca2+ current characteristics in endo- and epicardial myocytes from rat left ventricle with pressure-induced hypertrophy

Cardiac hypertrophy alters the regional distribution of action potential duration by affecting the magnitude and kinetics of underlying ionic currents. In the present study, the effect of pressure-induced cardiac hypertrophy on the magnitude and kinetics of the L-type Ca2+ current ( I(CaL)) was investigated in endo- and epicardial myocytes of the rat left ventricular free wall. Cardiac hypertrophy was induced by stenosis of the ascending aorta (AS), which led to a pressure difference between the left ventricular peak pressure and the peripheral systolic blood pressure of 84+/-6 mmHg ( n=5, P<0.0001). Sham-operated animals served as controls ( n=7). I(CaL) was investigated using the whole-cell patch-clamp technique. The magnitude of I(CaL) as well as its inactivation, steady-state activation, steady-state inactivation and recovery from inactivation were similar in endo- and epicardial myocytes of sham-operated rats. In myocytes isolated from AS animals, I(CaL) magnitude was significantly greater than in sham-operated animals (2407+/-127 pA, n=42 versus 1904+/-102 pA, n=33; P<0.01). This difference disappeared, however, when I(CaL) was normalized to cell capacitance. The kinetic properties of I(CaL) were unaffected by AS in both endo- and epicardial myocytes. In conclusion, in pressure-induced cardiac hypertrophy the density and kinetics of I(CaL) are preserved. This suggests that an additional insertion or recruitment of channels normally underlying I(CaL) into the cell membrane compensates for the increased cell size associated with hypertrophy.     

Biocompatibility of the polyurethane resin of the castor bean inserted into the alveolar bone of the dog

The biocompability of the polyurethane resin of the castor bean (Ricinus vulgaris) was studied following its insertion into the alveolar bone of dogs, after extraction of their premolar teeth. The resin was left to polymerise in the dental alveolus. Excess of material due to polymerisation was removed and polishing was employed to smooth and adapt the occlusal surface to the margins of the alveolar bone. This allowed a perfect suture of the mucosa together with the periosteum. The resin remained in the dental alveolus for 90 days. It was observed that the polyurethane was replaced by osteoid and bone tissues and no immune or inflammatory reactions were detected. There has been work on and discussion about the use of the polyurethane in grafts, prostheses and orthoses. Attention was paid to all the surgical steps, in particular the preservation of the periosteal integrity. Further research is being followed in our Department in order to test the biocompatibility of the material presented in this paper when used together with metallic or ceramic implants.     

Micromanipulation of single cells from tissue imprints is an alternative to laser-assisted microdissection

The characterization and analysis of single cells by molecular biological methods such as the polymerase chain reaction (PCR) is of increasing interest in biomedical research. Different techniques have been developed to obtain single cells from solid tissue. Currently, the most frequently used technique is laser-assisted microdissection (LAM). However, LAM of tissues cannot exclude contamination of the targeted cells by underlying cell fragments. Moreover, this technique can only be performed if a laser microscope is available. Thus, we developed a method to obtain single cells of fresh solid tissue by the simple technique of tissue imprints. After immunostaining of the imprints, single cells were transferred to a reaction tube using a 27-gauge needle guided by a mechanical micromanipulator. Consequently, we used these cells in a single cell PCR.     

RP59, a marker for osteoblast recruitment, is also detected in primitive mesenchymal cells, erythroid cells, and megakaryocytes.

We recently described a novel protein in bone marrow of rats, RP59, as a marker for cells with the capacity to differentiate into osteoblasts. In this work, its expression pattern was further investigated to learn about the origin and biological relevance of RP59 expressing marrow cells. As revealed by in situ hybridization and by immunohistochemistry of yolk sac embryos, RP59 was found in the cells of the primitive ectoderm and primitive streak as well as in blood islands and extraembryonal mesoderm. Later, RP59 occurred in fetal liver cells and in circulating blood. From the time around birth, it was found in bone marrow and spleen cells. In addition, in vitro-formed blood vessels contained RP59-positive cells in the lumen. Endothelial cells and the vast majority of cells outside the blood vessels were not labeled. Concerning more mature hematopoietic cell types, RP59 was observed in megakaryocytes and nucleated erythroblasts, but absent from lymphoid cells. In conclusion, RP59 was induced in early mesoderm. It was maintained in the erythroid and megakaryotic lineages and, as earlier described, in young osteoblasts.     

Severe rhabdomyolysis, acute renal failure and posterior encephalopathy after 'magic mushroom' abuse.

We report the case of a 25-year-old, hepatitis C-infected man, who presented with severe rhabdomyolysis and acute renal failure, and later developed posterior encephalopathy with cortical blindness after the ingestion of magic mushrooms. Conventional respiratory and cardiovascular support including mechanical ventilation, continuous veno-venous hemodialysis and corticosteroids led to improvement and the patient recovered completely over the following months. Magic mushrooms are becoming increasingly fashionable among drug users, as they are believed to be more harmless than other hallucinogenic designer drugs. So far, little is known about their possible severe side effects.     

The human error: delayed diagnosis of intravascular loss of guidewires for central venous catheterization

We report three recent cases in which there was intravascular loss of the entire guide wire. In one case, this loss was discovered more than three months after the patient's procedure. In another case, the loss was detected one month later, and in the third case, four days was the interval of the loss. We discuss the problems of human error and failure of diagnosis, both of which were responsible for serious sequelae.     

Death caused by strongyloides hyperinfection in a leprosy patient on treatment for a type II leprosy reaction

Strongyloides stercoralis is present worldwide and can cause hyperinfection in patients on long-term immunosuppressive doses of steroids, as is sometimes the case for patients treated for leprosy reactions. Strongyloides hyperinfection can present with ileus, as is discussed in this case report. Physicians, including surgeons, should be aware of this entity in order to avoid an unnecessary laparotomy. Though patients may survive if diagnosed at an early stage, strongyloides hyperinfection syndrome has a mortality rate of 87% and prevention is therefore of utmost importance.