Detection of Proteins in Normal and Alzheimer's Disease Cerebrospinal Fluid with a Sensitive Sandwich Enzyme-Linked Immunosorbent Assay

Abstract— Alzheimer's disease is a progressive degenerative dementia characterized by the abundant presence of neurofibrillary tangles in neurons. This study was designed to test whether the microtubule-associated protein, a major component of neurofibrillary tangles, could be detected in CSF. Additionally, we investigated whether CSF levels were abnormal in Alzheimer's disease as compared with a large group of control patients. We developed a sensitive sandwich enzyme-linked immunosorbent assay using AT120, a monoclonal antibody directed to human, as a capturing antibody. With this technique, the detection limit for was less than 5 pg/ml of CSF. Using ATS, which recognizes abnormally phosphorylated ser-ines 199–202 in, the detection limit was below 20 pg/ml of CSF. However, with AT8, we found no immunoreactiv-ity in CSF, suggesting that only a small fraction of CSF contains the abnormally phosphorylated AT8 epitope. Our results indicate that CSF levels are significantly increased in Alzheimer's disease. Also, CSF levels in a large group of patients with a diversity of neurological diseases showed overlap with CSF levels in Alzheimer's disease.     

PEAK SHIFTS AND POLYMORPHISM DURING PHASE THREE OF WRIGHT'S SHIFTING-BALANCE PROCESS

The third phase of Wright's shifting-balance theory involves the export of adaptive gene combinations from one subpopulation to another. Previous results have demonstrated that this can occur at very low migration rates, but it has been argued that this simply reflects the ability of migration to overcome selection and fix any (even deleterious) alleles. Here, previous analyses are extended by concentrating on the critical balance between forward and reverse migration rates that still allows phase III to proceed. It is shown that selective advantage, dominance, recombination rate, and the number of loci all affect the ability of a genotype to invade and become fixed in a new subpopulation, but it is unlikely that phase III will occur in the absence of differential migration unless the invading genotype consists of a few dominant loci with a large selection advantage, spreading into a few populations of lower fitness. Therefore, as was envisioned by Wright, differential migration from more to less fit populations will be necessary for phase III to occur under most circumstances.     

Reactions Catalyzed by Peg-Modified α-Chymotrypsin in Organic Solvents. Influence of Water Content and Degree of Modification

-Chymotrypsin was modified with cyanuric chloride activated monomethoxypolyethylene glycol (MPEG) with molecular weights 1900 and 5000. Using the higher molecular weight MPEG a product that was soluble in benzene at moderate levels of modification was obtained, whereas with MPEG 1900 almost all the enzyme's amino groups had to be modified for dissolving the conjugate. The catalytic activity decreased with increasing degree of substitution. Apparent V_max was considerably higher for the less modified enzyme preparation than for the more modified one, while K_m,app stayed almost constant. The modified enzyme was used for peptide synthesis. The reaction was dependent on the content of dissolved water. Both V_max,app and K_m,app increased with increasing water content. It was possible to achieve a process with complete conversion of substrate to dipeptide.     

The effect of salinity on the reproduction of coastal submerged macrophytes in experimental communities

The effects of salinity on the reproduction of coastal submerged macrophyte species were studied on samples of communities from six seasonal marshes in two outdoor experiments performed in autumn and in spring. The submerged macrophyte communities were submitted to five different salinity levels (0, 1, 2, 4 and 6 g/1 Cl^?1). In a companion paper (Grillas, van Wijck & Bonis 1993) three groups of species were distinguished on the basis of their biomass production over the salinity range 0 to 6 g/1 Cl^?1: (1) glycophytes (non-salt-tolerant species), (2) salt-tolerant species and (3) halo-phytes. This part of the study describes the impact of salinity on the reproduction of the individual species during the two experiments. The species differ in their capacity to reproduce in the autumn; only Zannichelliapedunculata and Tolypella hispánica were able to produce fruits in that season. For all species reproduction was greater in spring and strongly correlated with biomass, except for Chara canescens. Differences in reproductive effort over the salinity range amplified the halophytic nature of Ruppia marítima and Chara canescens and the intolerance of Callitriche truncata and Chara contraria. For the other species, reproductive effort did not differ significantly over the salinity range. Regarding the effect of salinity on biomass and reproductive effort of individual species, there were large differences in the total weight of propagules produced at the community level and in the relative contribution of individual species. The resulting quantitative changes in the species composition of the seed bank could affect the structure of the communities by their effects on the establishment and survival of species populations.     

Distribution,causal agents,and infection dynamic of emerging ink disease of sweet chestnut in Southern Switzerland

Emerging diseases caused by both native and exotic pathogens represent a main threat to forest ecosystems worldwide. The two invasive soilborne pathogens Phytophthora cinnamomi and Phytophthora × cambivora are the causal agents of ink disease, which has been threatening Castanea sativa in Europe for several centuries and seems to be re-emerging in recent years. Here, we investigated the distribution, causal agents, and infection dynamics of ink disease in southern Switzerland. A total of 25 outbreaks were identified, 19 with only P. cinnamomi, 5 with only P. × cambivora, and 1 with both species. Dendrochronological analyses showed that the disease emerged in the last 20–30 years. Infected trees either died rapidly within 5–15 years post-infection or showed a prolonged state of general decline until death. Based on a generalized linear model, the local risk of occurrence of ink disease was increased by an S-SE aspect of the chestnut stand, the presence of a pure chestnut stand, management activities, the proximity of roads and buildings, and increasing annual mean temperature and precipitation. The genetic structure of the local P. cinnamomi population suggests independent introductions and local spread of the pathogen.     

Henipaviruses and lyssaviruses target nucleolar treacle protein and regulate ribosomal RNA synthesis

The nucleolus is a common target of viruses and viral proteins, but for many viruses the functional outcomes and significance of this targeting remains unresolved. Recently, the first intranucleolar function of a protein of a cytoplasmically-replicating negative-sense RNA virus (NSV) was identified, with the finding that the matrix (M) protein of Hendra virus (HeV) (genus Henipavirus, family Paramyxoviridae) interacts with Treacle protein within nucleolar subcompartments and mimics a cellular mechanism of the nucleolar DNA-damage response (DDR) to suppress ribosomal RNA (rRNA) synthesis. Whether other viruses utilise this mechanism has not been examined. We report that sub-nucleolar Treacle targeting and modulation is conserved between M proteins of multiple Henipaviruses, including Nipah virus and other potentially zoonotic viruses. Furthermore, this function is also evident for P3 protein of rabies virus, the prototype virus of a different RNA virus family (Rhabdoviridae), with Treacle depletion in cells also found to impact virus production. These data indicate that unrelated proteins of viruses from different families have independently developed nucleolar/Treacle targeting function, but that modulation of Treacle has distinct effects on infection. Thus, subversion of Treacle may be an important process in infection by diverse NSVs, and so could provide novel targets for antiviral approaches with broad specificity.     

Transforming a set of biological flat file librariesto a fast access network

SRS (Sequence Retrieval System), an indexing system for flatfile libraries, provides fast access to individual library entriesvia retrie by keywords from rious data fields. SRS is now alsoable to build indices using cross–references that mostlibraries provide. Fifteen libraries of DNA and protein sequencesand structures have been selected. These libraries interactwith at least one other by means of cross–references.Indexing these cross–references allows a complete networkof libraries to be built. In the network an entry from one librarycan be linked in principle to every other library. If two librariesare not directly cross–referenced, the linkage can bemade with a succession of single links between neighbouring,cross–referenced libraries. A new operator has been addedto the query language of SRS for convenient specification oflinks amongst complete libraries or entry sets generated byprevious queries on particular libraries. All the informationin the network can now be used to retrieve an entry in a specificlibrary, e.g. the full information given in amino acid sequenceentries from SwissProt can now be used to retrieve related tertiarystructure entries from PDB. Furthermore, a search in a singlelibrary can be extended to a search in the complete librarynetwork, e.g. all entries in all databases pertaining to elastasecan be found.     

Selenium in combination with a tomato lipid extract as a therapy for benign prostatic hyperplasia and its alterations in rats with induced BPH

Benign prostatic hyperplasia (BPH) is the most common adenoma in old men. Tomatoes are a rich source of bioactive compounds that, as well as selenium (Se), possess antioxidant and antiproliferative activity. The aim was to evaluate the therapeutic effect of Se in combination with a tomato extract in aged rats with BPH. Aged male Wistar rats were divided in the following groups (n = 10 rats/group): Control (C), BPH, BPH + Finasteride (BPH + F), BPH + Tomato Lipidic Extract (BPH + E), BPH + Selenium (BPH + S) and BPH plus E plus S (BPH + E + S). After 4 weeks of treatment, prostate weight, diuresis, antioxidants enzymes, prooxidants and inflammatory markers, growth factors and androgens were determined. BPH + E + S reduced prostate weight by 59.29% and inhibited growth by 99.35% compared to BPH + F which only decreased weight and inhibited growth by 15.31% and 57.54%, respectively. Prooxidant markers were higher with BPH + F (49.4% higher vs. BPH), but BPH + E + S decreased these markers (94.27% vs. BPH) and increased antioxidant activity. Finally, diuresis was higher with the BPH + E + S combination and markers of inflammation and growth factors were significantly lower with respect to BPH + F. Our findings provide a beneficial and protective therapeutic option of E + S directed against androgens, oxidative stress and inflammation that regulates cell proliferation in the prostate gland.     

Engineered nickel bioaccumulation in Escherichia coli by NikABCDE transporter and metallothionein overexpression

Mine wastewater often contains dissolved metals at concentrations too low to be economically extracted by existing technologies, yet too high for environmental discharge. The most common treatment is chemical precipitation of the dissolved metals using limestone and subsequent disposal of the sludge in tailing impoundments. While it is a cost-effective solution to meet regulatory standards, it represents a lost opportunity. In this study, we engineered Escherichia coli to overexpress its native NikABCDE transporter and a heterologous metallothionein to capture nickel at concentrations in local effluent streams. We found the engineered strain had a 7-fold improvement in the bioaccumulation performance for nickel compared to controls, but also observed a drastic decrease in cell viability due to metabolic burden or inducer (IPTG) toxicity. Growth kinetic analysis revealed the IPTG concentrations used based on past studies lead to growth inhibition, thus delineating future avenues for optimization of the engineered strain and its growth conditions to perform in more complex environments.