Impact of endogenous nitric oxide on microglial cell energy metabolism and labile iron pool

Microglial activation is common in several neurodegenerative disorders. In the present study, we used the murine BV-2 microglial cell line stimulated with gamma-interferon and lipopolysaccharide to gain new insights into the effects of endogenously produced NO on mitochondrial respiratory capacity, iron regulatory protein activity, and redox-active iron level. Using polarographic measurement of respiration of both intact and digitonin-permeabilized cells, and spectrophotometric determination of individual respiratory chain complex activity, we showed that in addition to the reversible inhibition of cytochrome-c oxidase, long-term endogenous NO production reduced complex-I and complex-II activities in an irreversible manner. As a consequence, the cellular ATP level was decreased in NO-producing cells, whereas ATPase activity was unaffected. We show that NO up-regulates RNA-binding of iron regulatory protein 1 in microglial cells, and strongly reduces the labile iron pool. Together these results point to a contribution of NO derived from inflammatory microglia to the misregulation of energy-producing reactions and iron metabolism, often associated with the pathogenesis of neurodegenerative disorders.     

New in vivo and ex vivo models for the experimental study of sheep scrapie: development and perspectives

Sheep scrapie is a prototypical transmissible spongiform encephalopathy (TSE), and the most widespread of these diseases. Experimental study of TSE infectious agents from sheep and other species essentially depends on bioassays in rodents. Transmission of natural sheep scrapie to conventional mice commonly requires one or two years. In an effort to develop laboratory models in which investigations on the sheep TSE agent would be facilitated, we have established mice and cell lines that were genetically engineered to express ovine PrP protein and examined their susceptibility to the infection. A series of transgenic mice lines (tgOv) expressing the high susceptibility allele (VRQ) of the ovine PrP gene from different constructs was expanded. Following intracerebral inoculation with natural scrapie isolates, all animals developed typical TSE neurological signs and accumulated abnormal PrP in their brain. The survival time in the highest expressing tgOv lines ranged from 2 to 7 months, depending on the isolate. It was inversely related to the brain PrP content, and essentially unchanged on further passaging. Ovine PrP transgene expression thus enhanced scrapie disease transmission from sheep to mice. Such tgOv mice may bring new opportunities for analysing the natural variation of scrapie strains and measuring infectivity. As no relevant cell culture models for agents of naturally-occurring TSE exist, we have explored various strategies in order to obtain stable cell lines that would propagate the sheep agent ex vivo without prior adaptation to rodent. In one otherwise refractory rabbit epithelial cell line, a regulable expression of ovine PrP was achieved and found to enable an efficient replication of the scrapie agent in inoculated cultures. Cells derived from sheep embryos or from tgOv mice were also used in an attempt to establish permissive cell lines derived from the nervous system. Cells engineered to express PrP proteins of a specified sequence may thus represent a promising strategy to further explore, at the cellular level, various aspects of TSE diseases.     

Dementia,the fate of brain? Neuropathological point of viewLe destin du cerveau ? Un point de vue neuropathologique sur les démences.

The prevalence of dementia dramatically increases during ageing, and this puts a serious strain on the optimism brought by the continuous increase in life expectancy observed in most industrialised countries. Diseases that produce dementia are numerous, and the cognitive deficit results from lesions of various regions and from different mechanisms. This modulates the possible prediction, prevention and cure of dementia. Emphasis is put on the necessity of, and prerequisite for, efficient research in the field of dementia. Three paradigmatic dementing disorders are reviewed. Subacute spongiform encephalopathies (prion diseases) constitute a biological enigma and a public health concern. In Alzheimer's disease and vascular or mixed dementia, the clinical diagnosis is still imperfect, and this hinders research. Distinguishing and accurately identifying the various types of dementia is essential for understanding their mechanism and for developing efficient therapeutic strategies, preventive and curative. For such objectives, the study of human brain tissue will remain mandatory until non-invasive markers and additional models are available. Ethical reasons banish the use of cerebral biopsy and favour the promotion of autopsy.     

Gene expression profiling of human diseases by serial analysis of gene expression

Until recently, the approach to understanding the molecular basis of complex syndromes such as cancer, coronary artery disease, and diabetes was to study the behavior of individual genes. However, it is generally recognized that expression of a number of genes is coordinated both spatially and temporally and that this coordination changes during the development and progression of diseases. Newly developed functional genomic approaches, such as serial analysis of gene expression (SAGE) and DNA microarrays have enabled researchers to determine the expression pattern of thousands of genes simultaneously. One attractive feature of SAGE compared to microarrays is its ability to quantify gene expression without prior sequence information or information about genes that are thought to be expressed. SAGE has been successfully applied to the gene expression profiling of a number of human diseases. In this review, we will first discuss SAGE technique and contrast it to microarray. We will then highlight new biological insights that have emerged from its application to the study of human diseases.     

Ticks and Tick-Borne Diseases of Livestock Belonging to Resource-poor Farmers in the Eastern Free State of South Africa

The paper provides a summary of three studies conducted in the eastern Free State of South Africa between 1998 and 2000. In a questionnaire-based study approximately 21% of interviewed resource-poor farmers (n = 150) indicated that they experienced problems with ticks and tick-borne diseases. About 56% of farmers indicated that tick-related problems were most severe in summer, while 32% indicated that the most problems were encountered in winter. About 12% indicated that the tick problems were experienced throughout the year. Farmers also indicated that the highest tick burdens were experienced between spring and early winter. The principal ticks infesting cattle (n = 30) were found to be Boophilus decoloratus (53.1%), Rhipicephalus evertsi evertsi (44.7%), Rhipicephalus follis (1.0%), Rhipicephalus gertrudae (0.7%) and Rhipicephalus warburtoni (0.4%). On small stock (n = 188), R. evertsi evertsi (68%) and B. decoloratus (32%) were recorded as the main ticks in the study area. A sero-epidemiological survey of cattle (n = 386) showed that 94% of cattle were sero-positive for Babesia bigemina by IFAT, while 87% were sero-positive for Anaplasma by indirect ELISA. All the animals were sero-negative for Babesia bovis and this is probably because the tick vector, Boophilus microplus, is not present in the study area. All sheep and goats were sero-positive for Theileria species by IFAT while 85% of sheep and 100% of goats tested positive for Anaplasma species by competition inhibition ELISA. The high incidence of positive serological results for B. bigemina and Anaplasma in cattle, and Theileria and Anaplasma in sheep and goats and the absence of clinical cases would indicate that this area is endemically stable for these diseases. This revised version was published online in July 2006 with corrections to the Cover Date.     

The role of heme oxygenase signaling in various disorders

Modern methods of cell and molecular biology, augmented by molecular technology, have great potential for helping to unravel the complex mechanisms of various diseases. They also have the potential to help us try to dissect the events which follow the altered physiological conditions. Thus, there is every reason to believe that some of the potential mechanisms will be translated sooner or later into the clinic. Heme oxygenase (HO)-related mechanisms play an important role in several aspects of different diseases. In the past several years, significant progress has been made in our understanding of the function and regulation of HO. The objective of this article is to review current knowledge relating to the importance of HO mechanism in various diseases including myocardial ischemia/reperfusion, hypertension, cardiomyopathy, organ transplantation, endotoxemia, lung diseases, and immunosuppression. The morbidity and mortality of these diseases remain high even with optimal medical management. Furthermore, in this review, we consider various factors influencing the HO system and finally assess current pharmacological approaches to their control.     

The importance of microscopic examination in the management of desquamative diseases of the scalp

After determining the usual malassezic biota of the scalp in adult, normal persons, 259 patients with different desquamative diseases were studied by a simple adhesive tape technique. The main purpose of this study was to investigate the utility of this technique to improve the diagnosis and treatment of patients. Most patients with seborrhoeic dermatitis and sebopsoriasis demonstrated large numbers of {it Malassezia} spp. cells corresponding to the so called ``pityrosporosis'. Only 43.6% of patients with pityriasis capitis (dandruff) presented with such a diagnosis. Symptomatic pityrosporosis of the scalp should be treated with imidazolic derivatives or other antifungal substances. Patients with psoriasis of the scalp showed a typical microscopic picture represented by parakeratosic (nucleated) keratinocytes with absence of lipophilic yeasts which should be attributed to the usual dryness of the scales .Microbial epidermitis (eczema) of the scalp revealed another characteristic picture constituted by abundant leukocytes and bacteria without the presence of yeasts. The different microscopic pictures seen with this simple technique for diseases of the scalp, offer an adjunct to make a proper diagnosis and to establish a convenient treatment in cases which are not clinically well defined. This revised version was published online in June 2006 with corrections to the Cover Date.     

IL-15 up-regulates iNOS expression and NO production by gingival epithelial cells

To investigate the biological activity of epithelial cells in view of host defense, we analyzed the mRNA expression of inducible NOS (iNOS) as well as NO production by human gingival epithelial cells (HGEC) stimulated with IL-15. RT-PCR analysis revealed that HGEC expressed IL-15 receptor alpha-chain mRNA. In addition, stimulation with IL-15 enhanced iNOS expression by HGEC through an increase of both mRNA and protein levels. Moreover, IL-15 up-regulated the production of NO(2)(-)/NO(3)(-), a NO-derived stable end product, from HGEC. The enhanced NO production by IL-15 was inhibited by AMT, an iNOS-specific inhibitor. These results suggest that IL-15 is a potent regulator of iNOS expression by HGEC and involved in innate immunity in the mucosal epithelium.