To verify whether the use of potentially inappropriate medications (PIMs) is associated with loss of independence in elderly in-patients by promoting adverse drug reactions (ADRs).Prospective observational study.Five hundred six patients aged 65 and older admitted to 11 acute care medical wards.In-hospital loss of one or more activities of daily living (ADLs) and three or more ADLs. PIMs were identified according to diagnosis-independent Beers criteria and ascertained by study physicians based on daily review of medical and nurse records. The relationship between risk factors and outcomes was assessed using logistic regression.Overall, 104 patients (20.6%) were taking at least one PIM at the time of admission (baseline users), and 49 (9.7%) were newly prescribed at least one PIM during their hospital stay. The loss of one or more ADLs occurred in 9.6% of baseline users, 16.3% of new users, and 8.5% of nonusers (P=.21) and that of three or more ADLs in 7.7% of baseline users, 12.2% of new users, and 4.8% of nonusers (P=.10). The lack of association was confirmed after correction for potential confounders, including ADRs. The occurrence of ADRs was strongly associated with both outcomes (odds ratio (OR)=7.80, 95% confidence interval (CI)=3.53-17.3 for the loss of >= 1 ADLs; OR=3.98, 95% CI=1.50-10.5 for the loss of >= 3 ADLs), but PIMs caused only six of 106 ADRs.ADRs to any drugs more than the use of PIMs might be associated with functional decline in elderly hospitalized patients, but because the power of this study was too limited to definitively exclude a direct relationship between PIMs and functional decline, this merits further investigation.

Burt et al. (1) demonstrate that the APOE ε4/ε4 and other ε2+ or ε4+ genotypes accelerate HIV mortality, a gene dosage effect of the ε4 allele on steady-state viral load, and they associate ε4 with enhanced HIV-1 cell entry. They did not find an excess of HIV-associated dementia (HAD) among ε4+ patients. …

Background and aims - Alzheimer's disease (AD) is considered to be associated with a low-grade chronic neuroinflammation where activated glial cells produce pro-inflammatory cytokines, ISPs and Reactive Oxygen Species (ROS) that over long periods of time create a pro-oxidative state favouring the beta-amyloid (A beta) peptide fibrillation and aggregation leading to senile plaque formation and disease progression. Methods - We studied the effects of several ISPs (see table 1) on Ab25-35 induced fibrillar aggregation and cytotoxicity by: human Red Blood Cells, Lysis experiments and Neuroblastoma Cell Cultures - Apoptosis. Results - ISPs investigated were able to interact with the A beta-peptide but with differential effects and mechanisms on fibrillar aggregation and toxicity. Conclusions - Taking into account the results here reported and the previously demonstrated presence of ISPs in the senile plaques, it is possible to Tab. 1 Positive/negative inflammation sensitive proteins against Ab25-35 induced fibrillar aggregation and cytotoxicity [GRAPHICS] hypothesize an in vivo model where ISPs can act in a first phase as A beta-protecting or A beta-interacting substances that, however, in later phases may themselves be involved in the A beta aggregation process, thus explaining ISPs co-deposition together with A beta within the senile plaques.

objectives: A neuroinflammatory process, triggered by amyloid-beta (A beta)-peptide, is thought to play a central role in the neurodegencrative process leading to Alzheimer's disease (AD). A beta(25-35) retains the functionality of A beta(42) and was employed to investigate the effects of inflammation-sensitive proteins (ISPs) alpha l-antichyinotrypsin (AlACT) and alpha l-antitrypsin (AlAT) on fibrillar aggregation and cytotoxicity.Design and methods: Inhibitory concentrations of the ISPs were determined in an established human red blood cell lysis model of A[ cytotoxicity. For studies of A beta-fibrillar aggregation CSF levels of AlACT (0.041 mu M)/AlAT (0. 11 mu M) were incubated with Congo Red dye 25 mu M+ A beta(25-35) 10 mu M noting the formation of visible aggregates and spectrophotometrie changes over 24 h.Results: A I ACT at CSF reported levels inhibited fibrillar aggregation and cytotoxicity while A] AT at CSF reported levels failed to cause a similar inhibition.Conclusions: A1ACT neutralizes fibrillar aggregation and cytotoxicity of A beta-peptide more effectively than A1AT. Both proteins are known to be co-deposited with A beta within senile plaques of AD brains. (c) 2007 The Canadian Society of Clinical Chemists. Published by Elsevier Inc. All rights reserved.

Virus cell-to-cell spread has been reported for many different viruses and may contribute to pathogenesis of viral disease. The role played by cell-to-cell contact in hepatitis C virus (HCV) transmission was studied in vitro by cell cocultivation experiments. A human lymphoblastoid B-cell line, infected persistently with HCV in vitro (TO.FEHCV), was used as HCV donor [Serafino et al., 2003]; recipient cells were the human hepatoma HepG2 cell line. Both cell types were co-cultured for 48 hr to allow the cell-to-cell contacts. The hepatoma HepG2 cells are not permissive to free-virus infection, but they were infected successfully using TO.FEHCV cells as source of virus. The kinetics of viral RNA synthesis and the percentage of infected cells were compared in cell-mediated-and cell-free-viral infection. After co-cultivation, a consistent proportion of hepatoma cells replicated HCV and stably expressed viral antigens. Virus produced was infectious as demonstrated by the ability to reinfect fresh B-cells. This cell model shows that permissiveness to HCV infection can be achieved in vitro in non-permissive hepatoma cells by direct cell-to-cell contacts with infected human B-cells. This mechanism of virus spread may also play a pathogenic role in vivo.

Objectives: The aim of this study was to investigate transthyretin (prealbumin) effects oil A beta(25-35)-induced cytotoxicity.Design and methods: In view of the well-recognized literature data demonstrating that A beta(25-35) fibrillar aggregates cause in vitro cytotoxicity to human red blood cells and apoptotic changes to SK-N-BE neuroblastoma cells in cultures (ultrastructural evidence), we tested transthyretin effects on these two experimental models.Results: Incubation of A beta(25-35) with transthyretin (at transthyretin concentrations equal to CSF physiological levels) demonstrated both inhibition of red blood cells lysis and neutralization of SK-N-BE neuroblastoma cells ultrastructural apoptotic changes. Moreover, transthyretin was shown to be able to inhibit the formation of fibrillar macroaggregates of A beta(25-35).Conclusions: The findings imply that experimental systems investigating A beta-induced cytotoxicity consider the protective interaction of transthyretin with A beta; all interaction to be considered also in vivo in view of the fact that transthyretin immunoreactivity has been previously demonstrated in amyloid plaques of brains from Alzheimer's disease patients. (C) 2005 The Canadian Society of Clinical Chemists. All rights reserved.

Background: Fibrillar aggregates of amyloid beta 25-35 (Abeta(25-35)) form rapidly in vitro able to lyse human red blood cells (RBCs). Human sera, albumin, and apolipoprotein E (ApoE) each limit fibrillation and cytotoxicity. Potentially, these substances protect neurons from Abeta(1-40/42) aggregates. Transferrin (TF) is investigated in this study.Methods: The Mattson red blood cells model was employed to determine whether co-incubation of transferrin and Abeta(25-35) prevented lysis. The formation of fibrillar Abeta(25-35) in the presence of transferrin was investigated using Congo red staining and spectrophotometric studies.Results: We found that incubation of 20 muM Abeta(25-35) with physiologic levels of transferrin prevented red blood cells lysis and the formation of macro-aggregates.Conclusions: These in vitro results suggest that transferrin may limit fibrillar amyloid formation in vivo and cytotoxicity. (C) 2004 Published by Elsevier B.V.

The aim of this study was to inquire the antioxidant status in plasma and lipoproteins isolated from normal subjects possessing different ApoE genotypes. For this purpose we investigated blood samples from 106 healthy blood donors: the distribution of ApoE alleles (E2/E2 = 0.9%, E2/E3 = 10.4%, E2/E4 = 2.8%, E3/E3 = 71.7%, E3/E4 = 12.3% and E4/E4 1.9% with 1, 11, 3, 76, 13, and 2 subjects respectively for each genotype) was in agreement with previous data. Almost no differences were found in the concentrations of both coenzyme Q10 (CoQ10) and vitamin E for the different genotypes. Concentration of CoQ10 in isolated lipoproteins was also similar, in the different genotypes, when referred to cholesterol; CoQ10 in LDL was higher for the E3/E3 subjects when referred to protein. Neither CoQ10 nor vitamin E correlated with paraoxonase (PON) activity or cholesteryl-ester hydroperoxides (CHP). Furthermore, there was no correlation between the same lipophilic antioxidants and CHP levels. The only E2 homozygous subject found had high levels of PON and low levels of CHP; the two E4/E4 subjects had low PON activity together with low levels of CHP.

Abstract: In order to analyze the metabolic response of AD patient platelets to β‐amyloid, we have carried out fluorimetric measurements of intracellular calcium and an ultrastructural survey of platelets exposed to the β‐amyloid active fragment 25–35 (βA25‐35). Since it is not possible to analyze directly the damaged neurons in AD, the study of peripheral blood cells, especially platelets, may be of great value for the investigation of the toxic effects of β‐amyloid on AD neuronal cells.

Inhibition of the lysis of human red blood cells (RBCs) exposed to amyloid peptide Abeta25-35 is an model for screening natural and synthetic substances potentially protective against amyloid damage. In this system, human serum and a component, namely apolipoprotein E (apoE), completely prevent RBC lysis. This report demonstrates that albumin, another serum component, is 8-fold more protective: a concentration of 12.5 microg/ml protects RBCs against 20 microM-Abeta25-35, and prevents the formation of fibrillar Abeta25-35 aggregates stainable by Congo Red. The biological relevance of these findings is suggested by the following: (1) a large fraction ( approximately 90%) of circulating Abeta1-42 is bound to albumin; (2) albumin immunoreactivity is present in brain amyloid plaques; and (3) incubation of Abeta with albumin rapidly decreases detectable levels of free Abeta suggesting epitope masking. The results add new and important functional consequences to the amyloid-albumin relationship and imply that experimental systems investigating Abeta cytotoxicity should consider the protective interaction of albumin.

Mattson et al. (9) demonstrated lysis of human red blood cells (RBC) exposed to amyloid peptide Abeta(25-35), a new experimental model for amyloid-beta toxicity. Lysis resulted from poreformation in the RBC membranes and was completely prevented by concurrent exposure to Congo red We demonstrate that human serum, purified ApoE from human plasma, and recombinant isoforms of ApoE neutralize the Abeta(25-35) cytotoxicity: the E2 and E4 isoforms were marginally more effective than E3. Second, we demonstrate that Abeta(25-35) forms fibrils in the reaction mixtures using electron-microscopy. Together these results suggest that the RBC model might be useful in preliminary identification of natural and synthetic substances able to protect against amyloid-beta cytotoxic effects due to fibrillar Abeta(25-35). Such compounds would be candidate molecules for testing in neuronal systems.

Background Data in the literature have not clarified whether type 2 diabetes mellitus affects homocysteine plasma levels. Different variables able to influence homocysteine could be the cause of these controversial findings. An important but neglected confounding factor is Helicobacter pylori, which has been demonstrated to be a cause of elevated levels of homocysteine and which is prevalent in the Caucasian population, ranging from 30 to 40% incidence. Starting from these findings we wanted to verify whether differences in homocysteine levels exist between a type 2 diabetic population and a control group, taking into account the presence/absence of Helicobacter pylori.Design The study was carried out on a group of uncomplicated and normotensive type 2 diabetic patients (n = 30, 55.7 +/- 9.7 years) and on a control group (n = 43, 51.2 +/- 11.3 years). On these subjects we evaluated: main parameters of glyco- and lipometabolic balance, presence of Helicobacter pylori by C-13 Urea Breath Test, plasma homocysteine, vitamin B-12, folate and genetic polymorphism of methylenetetrahydrofolate reductase.Results Evaluating the two groups as a whole, significant differences in homocysteine were found when considering Helicobacter pylori presence/absence (14.0 +/- 6.5 vs. 10.6 +/- 4.7 mumol L-1, respectively, P < 0.01) without differences of vitamins and the genetic polymorphism. of methylenetetrahydrofolate reductase. The positive interaction found among Helicobacrer pylori, diabetes and homocysteine (P = 0.03) taking into account all the other evaluated confounding factors, demonstrates that a significant difference in homocysteine plasma levels exists between diabetics and controls (Helicobacter pylori-negative: diabetics 12.5 +/- 5.6 mu mol L-1, controls 9.4 +/- 38 mu mol L-1; Helicobacter pylori-positive: diabetics 13.6 +/- 5.8 mu mol L-1 controls 14.3 +/- 7.0 mu mol L-1).Conclusions Type 2 diabetes seems to induce per se higher levels of homocysteine, which appears to be one of the factors responsible for the increased risk of vascular damage.

Human aging is characterized by a complex remodeling of immune, endocrine, metabolic parameters.In particular, organismal aging is characterized by high levels of proinflammatory molecules paralleling a progressive decay of response to stress at cellular and systemic level The few data on centenarian fibroblasts suggest that these cells possess only some features of replicative senescence, indicating that in vitro senescence plays a role, but do not fully mimic in vivo senescence.On the whole, aging is more appropriately to be considered as the consequence of the remodeling of an expression pattern involving a great number of genes.To compare the pattern of gene expression in centenarians vs. young control fibroblasts, an array of 384 genes was designed and cDNAs were robotically spotted.Genes were chosen on the basis of relationship with cell senescence, role in cellular stress response, involvement in apoptosis, genes involved in inflammation, tumor suppressor genes, cell cycle regulators, genes whose products have been demonstrated to change during aging, and genes whose genetic variants have been associated with human longevity.

A form of beta-amyloid peptide A beta ending at amino acid 42 (A beta42) is the major component of senile amyloid plaques in Alzheimer's Disease (AD). The A beta-peptide earliest modifications are extremely important since they constitute the key events in the progression towards further changes finally leading to fibril formation and to A beta deposits which constitute the core pathological change in AD. Chemical and conformational early modifications of the beta-amyloid peptide are critical steps in AD pathogenesis and have been widely investigated. We now show that a Fenton-type OH-generating system is capable of generating L-Dopa (3,4-dihydroxyphenylalanine) in the tyrosine residue of A beta-peptide via aromatic ring hydroxylation, as the result of hydroxyl radical attack on proteins. Since L-Dopa is not a constituent of mammalian proteins and peptides, the formation of L-Dopa in A beta in vitro constitutes a possible important modification caused by hydroxyl radical attack. These results lay the groundwork for further studies on modification and damage associated with the degenerative disorder in AD where oxidative stress and inflammation are known to occur.

beta-amyloid (A beta) is a normal soluble peptide found in the cerebrospinal fluid (CSF) and other biological fluids. A beta fibrils are associated with Alzheimer's disease (AD) senile plaques. We have used purified soluble A beta (1-42) and A beta (12-28) peptides in order to determine the oxidative modification induced in these peptides by exposure to peroxidase and hydrogen peroxide. We have demonstrated that under these in vitro conditions, dimeric forms of A beta (1-42) can be detected by high-resolution polyacrylamide SDS-PAGE electrophoresis. Further experiments performed by reverse-phase high performance liquid chromatography (RP-HPLC), and monitored by fluorescence detection, showed that the dimeric A beta (1-42) forms induced by the peroxidase reaction are the outcomes of dityrosine bridge formation. This cross-link results from the enzyme catalyzed oxidation. During this reaction, phenolic coupling of tyrosine residues of two A beta (1-42) peptides occurs. No detectable peroxidative modifications were observed with the A beta (12-28) peptide which lacks a tyrosine residue. Since oxidative stress is thought to be associated with AD, the experimental model described here can help in understanding the early events leading to chemical, structural and conformational modifications before the conversion of sA beta to amyloid fibrils and eventually the formation of senile plaques in AD.

Alzheimer Disease (AD) is an age-related dementia that is going to assume the characteristics of a health emergency. A reliable diagnosis of AD, especially at the very onset, is complicated. Moreover, it is known that about 20% of patients in hospital for Alzheimer-type dementia have unusual clinical symptoms and sometimes a reliable diagnosis depends on post-mortem analysis of brain tissues. Recently, progress has been achieved in the field of genetic risk assessment in AD, and this article discusses the current widespread use of apolipoprotein E (APO E) genotyping as a tool for investigating epidemiologic risk factors for this devastating disease. APO E genotyping investigations have now moved from research laboratories to clinical laboratories, and therefore bioethical controversies are present, because of laboratory investigations of the patient's genome and the scientific uncertainties still existing. These controversies strongly suggest the need for genetic counseling. Herein we propose a model for multidimensional counseling provided by the Geriatric Assessment Unit (GAU), a multidisciplinary approach to counseling involving genetic, medical, social, psychological and medico-legal support. Multidimensional counseling may help patient and family decisions about health problems with the aim of improving the patient's information and knowledge, and ensuring informed consent in decisions concerning a disease associated with cognitive impairment.

The present study was aimed to decide whether Na+ influx can be involved in regulation of murine L 1210 leukemia cell growth. Cells were cultivated in the presence of different concentrations of amiloride and cellular growth was monitored by 3H-thymidine incorporation/105 cells. This drug inhibited cell growth in concentrations ranging from 1 x 10−5 to 1 x 10−3 mmol/ml. Even short time treatments with amiloride caused irreversible alterations: the cells, although survived, lost their ability to divide. The results support the hypothesis that Na+ influx is necessary for the duplication of tumor cells.