Intracellular sensing of microbes and danger signals by the inflammasomes

The cells of the innate immune system mobilize a coordinated immune response towards invading microbes and after disturbances in tissue homeostasis. These immune responses typically lead to infection control and tissue repair. Exaggerated or uncontrolled immune responses, however, can also induce acute of chronic inflammatory pathologies that are characteristic for many common diseases such as sepsis, arthritis, atherosclerosis, or Alzheimer's disease. In recent years, the concerted efforts of many scientists have uncovered numerous mechanisms by which immune cells detect foreign or changed self-substances that appear in infections or during tissue damage. These substances stimulate signaling receptors, which leads to cellular activation and the induction of effector mechanisms. Here, we review the role of inflammasomes, a family of signaling molecules that form multi-molecular signaling platforms and activate inflammatory caspases and interleukin-1β cytokines.     

Cerebral embolic ischemia in rats: correlation of stroke severity and functional deficit as important outcome parameter

The embolic MCA occlusion model in rats is used for recanalisation studies in acute stroke. In addition to the determination of lesion size, the assessment of functional outcome may improve the value of this model. Male Wistar rats were submitted to MCA clot embolism or sham surgery. In order to achieve a larger variety of lesion volume, 2 subgroups (each 7 animals) were subjected to differently sized emboli (30 and 40 mm). Follow-up period was 6 days. Outcome assessment consisted of a test battery including parallel bar crossing, observation of behaviour in an open field and an 8-arm maze and a neurological score with ten different sensorimotor and coordinative items. Animals were perfusion-fixed on day 7 (blinded examination). For both subgroups, there were significant impairments with regard to performance on the Neuro score, parallel bar crossing and maze exploration. Improvement was only partial during the follow-up period. On follow-up day 6, there was still a significant correlation between total infarct volume and functional outcome on the Neuro score (R=0.80, p=0.0006) and the exploration behaviour in the maze (R=0.66, p=0.01). Application of emboli with a length of 40 mm caused more functional impairment and a more extended lesion volume compared with 30 mm. We present outcome tests that provide quantitative and objective tools to test functional impairment in rats following embolic stroke.     

Population based quantification of dendrites: evidence for the lack of microtubule-associate protein 2a,b in Purkinje cell spiny dendrites

The high molecular weight isoforms (a and b) of microtubule-associate protein 2 (MAP2a,b) are widely believed to be specific markers for neuronal somata and dendrites. We analyzed and quantified MAP2a,b stained dendrites of the cerebellar molecular layer using a novel approach that segmented and 3D reconstructed them, and the results have been compared with those obtained by other methods, including single-cell reconstruction and analysis of electron micrographs. Our results show that the molecular layer dendritic volume fraction is lower than in the neocortex (10% compared to neocortical 29%). The low total volume fraction of dendrites in the molecular layer is best explained by the majority of the afferents to the dendrites being from the very densely packed parallel fibers, which allows the dendritic fields of individual neurons to be smaller and more compact than in the cerebral cortex. However, the MAP2a,b dendritic volume fraction is even lower (5.2%) than the total volume fraction of dendrites in the molecular layer (10%). Analysis of the material shows that this difference between the two results is due to the unexpected finding that there were few MAP2a,b stained Purkinje cell spiny dendrites.     

Analogs of GnRH-I and GnRH-II inhibit epidermal growth factor-induced signal transduction and resensitize resistant human breast cancer cells to 4OH-tamoxifen

About 50-64% of human breast cancers express receptors for GnRH-I. Direct antiproliferative effects of analogs of GnRH-I on human breast cancer cell lines have been shown. They are at least in part mediated by antagonizing growth promoting effects of estradiol, epidermal growth factor (EGF) or insulin-like growth factor. Recently, expression of a putative receptor for GnRH-II in human tissues was demonstrated. Antiproliferative effects of GnRH-II in human endometrial and ovarian cancer cells were shown not to be mediated through the GnRH-I receptor. Now we demonstrate direct anti-proliferative effects of the GnRH-I analog Triptorelin and the GnRH-II analog [d-Lys(6)]GnRH-II in MCF-7 and T47D human breast cancer cells expressing GnRH-I receptors and putative GnRH-II receptors. Pretreatment with Triptorelin or [d-Lys(6)]GnRH-II blocked EGF-induced autophosphoryla-tion of EGF receptor and activation of mitogen-activated protein kinase (extracellular-signal-regulated kinase 1/2 (ERK1/2)) in these cells. In sublines of MCF-7 and T47D cells, which were developed to be resistant to 4OH-tamoxifen, HER-2/p185 was overexpressed. Pretreatment of these cell lines with Triptorelin or [d-Lys(6)]GnRH-II completely abolished resistance to 4OH-tamoxifen, assessed by 4OH-tamoxifen-induced apoptosis. Analogs of GnRH-I and GnRH-II counteract EGF-dependent signal transduction in human breast cancer cells with expression of receptors for GnRH-I and GnRH-II. Through this mechanism, they probably reverse acquired resistance to 4OH-tamoxifen mediated through overexpression or activation of receptors of the c-erbB family.     

Cell distributions and functions of Toll-like receptor 4 studied by fluorescent gene constructs

Bacterial lipopolysaccharide (LPS) is recognized in mammals by a receptor complex composed of CD14, Toll-like receptor 4 (TLR4), and MD-2. The detailed mechanisms of how TLR4 transmits the signal from the outside to the inside of the cell remain to be elucidated. One way of studying TLR4 signaling mechanisms is to construct chimeras of TLR molecules C-terminally fused to fluorescent proteins and stably express these constructs in cells. Such constructs are functional when transfected into HEK293 epithelial cells. Confocal microscopy of TLR4 expression in live cells demonstrated pronounced expression on the plasma membrane as well in the Golgi apparatus. Studies were performed to clarify whether expression of TLR4 in the Golgi was necessary for LPS stimulation. Rapid recycling of TLR4/CD14/MD-2 complexes between the Golgi and the plasma membrane was a prominent phenomenon. In agreement with other types of plasma membrane receptors, aggregation of TLR4 by immobilized TLR4 antibodies was sufficient to induce signaling. Also, pharmacological disruption of the Golgi did not inhibit LPS induced NF-kappaB activation. Furthermore, LPS stimulation recruited the adapter molecule, MyD88, to the inside of the plasma membrane. Thus, LPS signaling commences on the plasma membrane and is independent of trafficking to the Golgi.     

Extracranial Internal Carotid Artery Occlusion: The Role of Common Carotid Artery Volume Flow

BACKGROUND AND PURPOSE: Quantitative measurement of blood flow volume in the common carotid artery (CCA) is now possible using the color velocity imaging quantification (CVI-Q) ultrasound technique. The aim of this study was to evaluate the cerebral hemodynamic effects of unilateral internal carotid artery (ICA) occlusion on CCA blood flow volumes (FVs) using CVI-Q. METHODS: Records of ultrasound studies in our neurosonology laboratory were retrospectively reviewed to identify patients with unilateral ICA occlusions who at a minimum received both a routine color duplex carotid ultrasound examination and quantitative measurement of FV in the CCA, bilaterally, using the CVI-Q method. A total of 71 patients met criteria and were included in the cohort. A side to side comparison was performed for FV, peak systolic velocities (PSV), end-diastolic velocities (EDV), and resistance indices (RIs) in the CCA. Results correlated with any other available data such as flow direction in the ophthalmic artery and the presence of intracranial collateralization. RESULTS: The FV, PSV, and EDV were significantly reduced, and the RI was significantly increased in the CCA on the side of the occlusion. A subgroup analysis in patients who also had an examination of the ophthalmic (n = 61) and the intracranial arteries of the Circle of Willis (n = 50), showed significantly higher FV in the CCA on the side of the occlusion if there was also reversed flow in the ophthalmic artery on the side of the occlusion (344 +/- 144 ml/min versus 169 +/- 53 ml/min). CONCLUSION: Quantitative FV measurement using CVI-Q ultrasound can identify clear alterations in volume flow, collateral pathways, and cerebral hemodynamics in patients with unilateral ICA occlusion. It is a complementary tool, providing additional objective information about the cerebral hemodynamic effects of ICA occlusion that goes beyond what is available using routine flow velocity data.     

Veränderungen im Chromosomensatz heterolog wachsender Tumoren

Zusammenfassung Der Chromosomensatz einer Walker-Carcinom-Linie, welche auf Ratten, Mäusen und Goldhamstern wächst, wurde untersucht. Die Morphologie der Chromosomen und ihre Zahl ist auf allen drei Tierarten in dieser Tumorlinie gleich. Ein auftretendes submetazentrisches Marker-Chromosom mit heterochromatischem Abschnitt findet sich in gleich hohem Prozentsatz in Tumor-zellen aller drei Tierarten.

---

Summary The chromosome-sets of a Walker carcinoma line were studied while growing in rats, mice, and gold hamsters. The morphology of the chromosomes and their number for this tumor-line were the same in all three types of animals. A submetacentric marker-chromosome appearing with heterochromatic segments was found in the same high percentage in the tumor cells of all three types of animal.

---

---

Mit 3 Textabbildungen

---

Der Deutschen Forschungsgemeinschaft danken wir für die Unterstützung bei Durchführung der Arbeit.