Acetyl Cholinesterase Inhibitors and Cell-Derived Peripheral Inflammatory Cytokines in Early Stages of Alzheimer's Disease.

BACKGROUND: Clinical and preclinical studies firmly support the involvement of the inflammation in the pathogenesis of Alzheimer's disease (AD). Despite acetylcholinesterase inhibitors (AChEI) being widely used in AD patients, there is no conclusive evidence about their impact on the inflammatory response. METHODS: This study investigates peripheral proinflammatory cytokines (interferon gamma [IFN-γ], tumor necrosis factor alpha [TNF-α], and interleukins 1ß [IL-1ß] and 6 [IL-6]) by firstly comparing peripheral blood mononuclear cell (PBMC)-derived secretion in drug-naïve and AChEI-treated AD patients versus healthy controls. A subset of those drug-naïve AD patients, who were prescribed the AChEI donepezil, was followed-up for 6 months to investigate if donepezil suppresses proinflammatory cell-derived cytokine secretion. RESULTS: Patients with AD showed higher levels of PBMC-derived proinflammatory cytokines (IFN-γ, TNF-α, IL-1ß, and IL-6) in comparison with healthy controls. On reexamination, previously drug-naïve AD patients who received donepezil treatment for 6 months displayed a decrease in cell-derived IFN-γ, TNF-α, IL-1ß, and IL-6. CONCLUSIONS: Proinflammatory PBMC-derived cytokines were increased in patients with AD in comparison with healthy controls and donepezil-reduced proinflammatory cytokines when examining drug-naïve AD patients before and after AChEI treatment.

Hydrogen bonds in protein-DNA complexes: where geometry meets plasticity.

Recognition of a DNA sequence by a protein is achieved by interface-coupled chemical and shape complementation. This complementation between the two molecules is clearly directional and is determined by the specific chemical contacts including mainly hydrogen bonds. Directionality is an instrumental property of hydrogen bonding as it influences molecular conformations, which also affects DNA-protein recognition. The prominent elements in the recognition of a particular DNA sequence by a protein are the hydrogen-bond donors and acceptors of the base pairs into the grooves of the DNA that must interact with complementary moieties of the protein partner. Protein side chains make most of the crucial contacts through bidentate and complex hydrogen-bonding interactions with DNA base edges hence conferring remarkable specificity.

The in vitro effective antiviral action of povidone-iodine (PVP-I) may also have therapeutic potential by its intravenous administration diluted with Ringer's solution.

The use of povidone-iodine (PVP-I) is well known in clinical medical practice. In vitro studies of cell cultures infected by HIV and H5N1 virus have shown that PVP-I has an antiviral action, while the cell hosts were not affected and survived. It is therefore worth investigating whether PVP-I, diluted with Ringer's solution, may have a therapeutic effect by parenteral administration. Specifically, the question is whether small concentrations of intravenous PVP-I could be well tolerated by the human organism, and in addition, if it would be possible to detect a beneficial activity. Its intravenous use may have a potential value against infections (by microbes, viruses, fungi and parasites), as well as an anti-inflammatory activity, especially in cases where antibiotics are ineffective. It could be used as a blood disinfectant, for treating burns, for the prevention of cancer, for the therapy of H5N1 influenza after its mutation, and other potential applications.

Application of denaturing gradient gel electrophoresis (DGGE) to screen for mutations of the human glucocorticoid receptor alpha gene (hGRalpha).

OBJECTIVES: In a previous publication, we had presented a sensitive method to detect mutations of the segment of the human glucocorticoid receptor alpha (hGRalpha) gene encoding the ligand binding domain (LBD) and part of the DNA binding domain (DBD) of hGRalpha, as several types of glucocorticoid resistance syndromes have been correlated with mutations in the respective nucleotide sequences. However, mutations affecting various regions covering the whole length of hGRalpha are increasingly reported in a variety of disease states. We now present an expanded screening methodology to detect mutations covering the whole length of hGRalpha. DESIGN AND METHODS: We developed a sensitive, simple screening PCR-DGGE method to detect mutations in the aminoterminal domain and DNA-binding domain of the hGRalpha. Wild type hGRalpha cDNA and mutant samples were included in the analysis to ensure the accuracy and sensitivity of the method. RESULTS: The PCR-DGGE method identified the mutant samples and discriminated them from wild type hGRalpha. CONCLUSIONS: The method described is accurate, sensitive, simple, cheap and fulfills the critera for a screening method which will be useful in delineating possible involvement of hGRalpha mutations in the aetiopathology of diseases correlated to derangements of glucocorticoid action.

Gene analysis of the glucocorticoid receptor alpha in Alzheimer's disease.

BACKGROUND: In Alzheimer's disease (AD), the hypothalamic-pituitary-adrenal (HPA) axis is hyperactive and the sensitivity to dexamethasone is decreased, suggesting a possible involvement of glucocorticoid receptor alpha (GRalpha) defects in the aetiopathology of the disease. METHODS: We, therefore, searched for the presence of mutations in the human GRalpha (hGRalpha) gene, focusing on the hormone-binding domain due to its importance in mediating glucocorticoids' effects. RNA isolated from peripheral blood mononuclear cells (PBMCs) of 15 patients with Alzheimer's disease and 20 healthy individuals was subjected to reverse transcription-polymerase chain reaction amplification (RT-PCR) analysis followed by denaturing gradient gel electrophoresis (DGGE). RESULTS: No mutations could be detected in the region of the hGRalpha gene examined. CONCLUSIONS: We conclude that the hormone-binding domain of GRalpha is not altered in Alzheimer's disease and molecular defects in other gene regions of the GRalpha or in its isoform GRbeta warrant further investigation in Alzheimer's disease.

Levels of serum cytokines and acute phase proteins in patients with essential and cancer-related thrombocytosis.

Essential thrombocytosis (ET) is a myeloproliferative disorder resulting in an increased production of abnormal platelets. Reactive thrombocytosis (RT) is occasionally observed in clinical situations including chronic inflammation and malignancy. The aim of the present study was to evaluate the discriminatory efficiency of various laboratory tests in patients with ET and cancer-related RT. Forty-five patients with ET, 52 patients with RT, and 25 age-matched normal individuals comprised the study population. Plasma interleukin-1 alpha (IL-1a), IL-2, IL-6, tumor necrosis factor alpha (TNF-a), platelets, hematocrit, hemoglobin, erythrocyte sedimentation rate (ESR), C-reactive protein (CRP), lactate dehydrogenase (LDH) and ferritin were determined. We found increased levels of ferritin, LDH, CRP, ESR, IL-1a, and IL-6 in RT compared with ET (p < 0.01 to p < 0.0005). Hemoglobin, hematocrit, and platelets were significantly lower in RT than in ET (p < 0.0005). Furthermore, ferritin and ESR were negatively correlated with Hct, hemoglobin, and TNF-a, whereas ferritin was positively correlated with ESR, IL-1a, IL-6, and CRP, and IL-1a was positively correlated with IL-6. We consider that the aforementioned parameters should be included in the investigation of unexplained thrombocytosis for the differentiation of essential from cancer related thrombocytosis.