Detection of chronic sensorimotor impairments in the ladder rung walking task in rats with endothelin-1-induced mild focal ischemia

A comprehensive evaluation of the effects of neuroprotection, neurogenesis, and compensatory mechanisms on the outcome of ischemic insults requires assessment of morphological and functional parameters. Behavioural tests are essential when recording performance throughout the time course of an experiment and the results bear predictive value in preclinical animal models. The goal of this study was to establish a behavioural test procedure for a model of transient focal ischemia induced by injection of endothelin-1 (eMCAO) that results in relatively mild behavioural deficits. The test protocol used in the present study allows evaluation of quantitative and qualitative impairments in skilled motor performance and is sensitive to detect chronic deficits at chronic post-ischemic time intervals. The ladder rung walking task [J. Neurosci. Methods 115 (2002) 169] is a motor test that assesses skilled walking and measures both forelimb and hindlimb placing, stepping and inter-limb co-ordination. In this study we tested the effect of two different technical variants of endothelin-1 application on infarct volume and motor skills (1) application via pre-implanted guiding cannula in awake animals and (2) via direct injection under halothane anaesthesia. We showed that the ladder rung walking task is sensitive in the assessment of loss of fine motor function after induction of relatively small lesions. In animals with implanted cannulas we found a smaller infarct area and an increase in placement errors prior to ischemia animals with eMCAO under anaesthesia showed a long lasting impairment of the contralateral forelimb up to 4 weeks post-eMCAO.     

Effects of a xanthine derivative, propentofylline, on local cerebral blood flow and glucose utilization in the rat

The effects of the xanthine derivative propentofylline [3-methyl-1-(5′-oxohexyl)-7-propylxanthinel were measured on local cerebral blood flow and glucose utilization in the rat using quantitative autoradiographic techniques. A dose of 0.5 mg/kg/min i.v. produced increases in local cerebral blood flow and minimal effects on glucose utilization in the majority of cerebral structures measured. A higher dose of propentofylline (1.5 mg/kg/min) produced an overall increase in local cerebral blood flow and a marked reduction in glucose utilization. Furthermore, propentofylline increased the average ratio of blood flow per unit glucose utilization and thus is capable of increasing cerebral blood flow in excess of metabolic demand. While the mechanism of action of this compound has not been fully defined. it is possible that its cerebrovascular and cerebral metabolic effects can at least partially be explained by a blockade of adenosine uptake. These actions of propentofylline on cerebral blood flow and metabolism may play a role in protecting neuronal tissue under hypoxic/ischemic conditions in the brain.     

Enhancement of glutathione and g-glutamylcysteine synthetase,the rate limiting enzyme of glutathione synthesis,by chemoprotective plant-derived food and beverage components in the human hepatoma cell line HepG2

Glutathione (GSH) is an important antioxidant and cofactor of detoxifying metabolism. Therefore, elevation of GSH as achieved by inducing g-glutamylcysteine synthetase (GCS), the limiting enzyme of GSH synthesis, may contribute to chemoprevention against cancer. In previous animal studies, increases in GCS were mainly found in liver and other organs that are not easily accessible in humans. Thus, employment and evaluation of alternative systems such as human-derived cell lines are encouraged. In the present experiment, we used the hepatoma cell line HepG2 to investigate the response of GCS and GSH to five plant-derived chemoprotectants contained in regularly consumed foodstuffs and beverages (kahweol/cafestol [K/C] [15.5-62.0 mM], a-angelicalactone [100-400 mM], benzyl isothiocyanate [1.7-5.0 mM], diallyl sulfide [175-700 mM], and quercetin [10-50 mM]). All treatments led to dose-dependent increases in both GCS activity and GSH concentration. Time course studies with K/C indicated that the enhancement of GCS preceded that of GSH, suggesting a causal relationship. K/C did not enhance g-glutamyl transpeptidase, a further enzyme that assists GSH-related chemoprotection. Although GCS induction has been suggested to require an initial short-lived GSH depletion, we did not find any decrease in GSH after 3 h of incubation with K/C. In summary, HepG2 cells were shown to be a useful model to investigate the capacity of potential chemoprotectants to enhance GCS and GSH. To our knowledge, the present study is also the first to show increases in GCS by K/C and a-angelicalactone in vitro and by diallyl sulfide and quercetin in any system.     

Anterior chamber angle measurement with optical coherence tomography: intraobserver and interobserver variability

PURPOSE: To assess intraobserver and interobserver variability of anterior segment optical coherence tomography (AS-OCT) as an objective diagnostic tool to quantify the anterior chamber angle and opening width. SETTING: University Eye Clinic, Lübeck, Germany. METHODS: The anterior chamber angle and opening width were assessed in 18 eyes of 9 healthy volunteers by 2 observers. Intraobserver reproducibility was evaluated by calculating an intraclass correlation coefficient (ICC) in a mixed model. Each observer had a separate model using information from 5 scans. Interobserver variability was determined by Bland-Altman analysis. The ICC was calculated in a mixed model using a residual maximum likelihood method. The results of 3 and 5 repeated scans were evaluated to indicate the change to 1 measurement application. RESULTS: The mean anterior chamber angle measurement was 35.9 degrees +/- 5.7 (SD) for observer A and 36.2 +/- 5.7 degrees for observer B. The ICC was 0.94 and 0.91, respectively. The mean opening width was 315 +/- 62 microm for observer A and 317 +/- 60 microm for observer B. The ICC was 0.97 and 0.93, respectively. Interobserver comparisons showed a mean difference between anterior chamber angle measurements of -0.27 +/- 1.6 degrees, a limit of agreement (LOA) interval from -3.52 to 2.98 degrees, and an ICC estimate of 0.96. The mean difference in opening width measurements was 2.40 +/- 12.40 microm, the LOA from -27.20 to 22.40 microm, and the estimated ICC 0.96. Using 1 instead of the mean of 5 measurements, the LOA range increased by 3.46 degrees for the anterior chamber angle and 30.0 microm for the opening width. CONCLUSION: Anterior chamber angle and opening width measurements by OCT showed low intraobserver and interobserver variability, indicating OCT is a valuable technique for quantitative assessment that provides reproducible measurements and objective documentation by different examiners.     

Is there an indication for high-dose chemotherapy in the treatment of bone and soft-tissue sarcoma?

Sarcomas represent a rare and heterogeneous disease, and the prognosis of patients with unresectable or advanced metastatic bone and especially soft-tissue sarcomas remains poor, with a disease-free survival of less than 10% at 5 years. Only few chemotherapeutic agents have been identified to be active, with reported response rates for doxorubicin, epirubicin and ifosfamide above 20%. Although combination therapy with different chemotherapeutic substances results in higher response rates, superiority against single-agent chemotherapy in terms of survival has not been demonstrated yet. Regarding anthracyclines, and ifosfamide in particular, a dose-response relationship has been shown, and high-dose chemotherapy with stem cell support has also been evaluated by several investigators. However, all studies comprised small patient cohorts and included very heterogeneous histological subtypes of soft-tissue sarcoma. Moreover, randomized trials are missing. Nevertheless, higher doses of chemotherapy result in higher response rates possibly correlating with longer survival. Finally, well-designed randomized trials should be performed, preferably in younger patients and in the context of an interdisciplinary treatment approach. In this review, we provide an overview of the literature concerning high-dose chemotherapy with hematopoietic stem cell support in the treatment of bone and soft-tissue sarcomas. On the basis of our own data, we would like to emphasize the importance of high-dose chemotherapy in the treatment of sarcomas.     

From HLA-B27 to spondyloarthritis: a journey through the ER

Summary: Almost four decades of research into the role of human leukocyte antigen-B27 (HLA-B27) in susceptibility to spondyloarthritis has yet to yield a convincing answer. New results from an HLA-B27 transgenic rat model now demonstrate quite convincingly that CD8⁺ T cells are not required for the inflammatory phenotype. Discoveries that the HLA-B27 heavy chain has a tendency to misfold during the assembly of class I complexes in the endoplasmic reticulum (ER) and to form aberrant disulfide-linked dimers after transport to the cell surface have forced the generation of new ideas about its role in disease pathogenesis. In transgenic rats, HLA-B27 misfolding generates ER stress and leads to activation of the unfolded protein response, which dramatically enhances the production of interleukin-23 (IL-23) in response to pattern recognition receptor agonists. These findings have led to the discovery of striking T-helper 17 cell activation and expansion in this animal model, consistent with results emerging from humans with spondyloarthritis and the discovery of IL23R as an additional susceptibility gene for ankylosing spondylitis. Together, these results suggest a novel link between HLA-B27 and the T-helper 17 axis through the consequences of protein misfolding and open new avenues of investigation as well as identifying new targets for therapeutic intervention in this group of diseases.