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International Journal of Alzheimer's Disease
Volume 2010, Article ID 548145, 12 pages
cNEUPRO: Novel Biomarkers for Neurodegenerative Diseases
1Laboratory for Molecular Neurobiology, Department of Psychiatry and Psychotherapy, University of Duisburg-Essen, LVR-Klinikum Essen, Virchowstraße 174, 45147 Essen, Germany
2Department of Psychiatry and Neurochemistry, Institute of Neuroscience and Physiology, The Sahlgrenska Academy at the University of Gothenburg, 431 80 Mölndal, Sweden
3Inserm U837, 1 Place de Verdun, 59045 Lille, France
4Faculte de Medicine, Université Lille-Nord de France, UDSL, rue Paul Duez, 59800 Lille, France
5CHU, bd. Pr J. Leclerc, 59037 Lille, France
6Department of Neurology, Institute of Clinical Medicine, University of Eastern Finland, Ylipistonranta 1C, 70211 Kuopio, Finland
7OncoProteomics Laboratory, Department of Medical Oncology, VU University Medical Center, De Boelelaan 1117, 1081 HV Amsterdam, The Netherlands
8Department of Neurology, University of Szeged, P.O. Box 427, 6701 Szeged, Hungary
9Department of Psychiatry and Psychotherapy, University of Erlangen, Schwabachanlage 6, 91054 Erlangen, Germany
10Functional Proteomics, Medizinisches Proteom-Center, Ruhr-University Bochum, Universitätsstraße 150, 44780 Bochum, Germany
11Medical Proteomics/Bioanalytics, Medizinisches Proteom-Center, Ruhr-University Bochum, Universitätsstraße 150, 44780 Bochum, Germany
12Medical Toxicology Centre, Institute for Ageing and Health, Institute of Neurosciences, University of Newcastle, Wolfson Unit, Claremont Place, Newcastle upon Tyne NE2 4AA, UK
13Department of Neurology, University of Ulm, Steinhövelstraße 1, 89075 Ulm, Germany
14Clinica Neurologica, Università di Perugia, Ospedale S. Maria della Misericordia, 06156 Perugia, Italy
15Department of Clinical Chemistry, VU University Medical Center, P.O. Box 7057, 1007 MB Amsterdam, The Netherlands
Received 14 April 2010; Accepted 5 July 2010
Academic Editor: Tuula R. M. Pirttila
Copyright © 2010 Philipp Spitzer et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
"clinical NEUroPROteomics of neurodegenerative diseases" (cNEUPRO) is a Specific Targeted Research Project (STREP) within the sixth framework program of the European Commission dedicated to the search for novel biomarker candidates for Alzheimer's disease and other neurodegenerative diseases. The ultimate goal of cNEUPRO is to identify one or more valid biomarker(s) in blood and CSF applicable to support the early and differential diagnosis of dementia disorders. The consortium covers all steps required for the discovery of novel biomarker candidates such as acquisition of high quality CSF and blood samples from relevant patient groups and controls, analysis of body fluids by various methods, and finally assay development and assay validation. Here we report the standardized procedures for diagnosis and preanalytical sample-handling within the project, as well as the status of the ongoing research activities and some first results.
1. Introduction
The diagnosis of Alzheimer's Disease (AD) is currently based primarily on clinical symptoms. Whereas the sensitivity of the clinical diagnosis for possible and probable Alzheimer Dementia according to National Institute of Neurological and Communicative Disorders and Stroke and the Alzheimer's Disease and Related Disorders Association (NINCDS-ADRDA) criteria is over 80%, its specificity is rather low [1]. The term mild cognitive impairment (MCI) was introduced for subjects who complain about verifiable cognitive disturbances but who show a preserved general cognitive functioning and no impairment in the activities of daily living [2]. These patients can be further subdivided into those with an impaired memory function (amnestic MCI) and those whose memory is preserved but who show disturbances of language, executive function, or visual-spatial skills (Nonamnestic MCI) [2]. If only one of the above-mentioned cognitive domains is impaired, patients are called single-domain MCI; if two or more domains are affected, they are referred to as multidomain MCI. Although the term MCI is solely descriptive and allows no conclusion on the aetiology, the classification allows some prediction of the course of the disease. For amnestic MCI patients, the risk to convert to Alzheimer's Dementia is 10–15% per year [3]. Yet, an accurate early diagnosis in MCI patients or even a predictive diagnosis in individuals without cognitive disturbances is still virtually impossible. As there is evidence that pathological biochemical changes start many years before the occurrence of functional symptoms, identification of biological markers in individuals with early-stage dementia is the most promising way to facilitate a predictive diagnosis [4–6].
Improving the early and predictive diagnosis of AD is of paramount importance if, in the future, preventive and disease-modifying therapies become available. In this regard, enormous efforts are under way. Although most therapies failed to show efficacy in Phase III trials, there are still some promising approaches like A lowering compounds, inhibitors of inflammation, inhibitors of tau phosphorylation and aggregation, and compounds interfering with cholesterol metabolism under investigation [7]. Although the brain has some limited regenerative capacity, neurons are still difficult to replace [8, 9]. Therefore, it is clear that maximal benefit for the patients can be expected when the treatment can be initiated as early as possible in the course of the disease. Furthermore, biologically valid and clinically accurate biomarkers may serve in the development of novel therapeutic strategies and may provide important information in clinical trials of therapies [10].
Well-documented biomarkers for AD in cerebrospinal fluid (CSF) include alterations in A, total-tau, and phospho-tau [10]. Importantly, these particular changes are detectable in early dementia stages as well as in individuals with mild cognitive impairment (MCI) who are at high risk of conversion to AD [11]. When analyzed in well-characterized clinical samples, the measurement of A, tau, and phospho-tau in cerebrospinal fluid generally allows the diagnosis of AD and even the prediction of the conversion from MCI to AD with a specificity and sensitivity of about 85% [12]. However, some report a lower sensitivity of below 50% for single biomarkers when these biomarkers are measured as part of a routine diagnostic test in a memory clinic [13]. This drop in sensitivity can be explained by the fact that in clinical practice the reference cohort is not a group of cognitively healthy individuals but consists of patients with other neurodegenerative and neurologic diseases who may also have slightly elevated total-tau, phospho-tau, or A levels [13]. The application of these markers in the differential diagnostic of neurodegenerative diseases therefore proves to be particularly problematic [14]. Consequently, there is a need for additional and more sensitive CSF biomarkers for the early and differential diagnosis of Alzheimer's Disease.
There is the additional problem of lumbar puncture to obtain CSF, since although the rate of complications during and after lumbar puncture is below 2–4% and restricted to mild to moderate postlumbar puncture headache [15–18], it must still be seen as invasive method for which special precautions must be taken. Consequently, there is a pressing need for new biomarkers in more easily accessible body-fluids such as peripheral blood.
Clinical proteomics is a fast developing field dedicated to the search for new biomarkers applicable to support the clinical diagnosis [19]. At present, a number of potential new biomarker-candidates for AD have been reported from proteomic studies [20, 21]; unfortunately, however, the published data is often contradictory and in many cases, a solid reassessment by other methods and with independent samples is required [19].
Taking this into account, the EU-project Clinical Proteomics for Neurodegenerative Diseases (cNEUPRO) is not only dedicated to the detection of potential new biomarker candidates for neurodegenerative diseases in CSF and blood, but also to the implementation of in-depth reassessments and validation studies. Finally, promising biomarker candidates will be studied for their suitability as routine test analytes by prototype assays.
2. cNEUPRO: The Consortium, Goals, and Workflow
cNEUPRO (http://www.cneupro.eu/) is a Specific Targeted Research Project (STREP) within the sixth framework program of the European Commission. It started in April 2007 and is coordinated by Jens Wiltfang, University of Duisburg-Essen. For the general aims of cNEUPRO, (see Box 1). The consortium consists of 14 academic partners (University of Duisburg-Essen, Centre Hospitalier Universitaire de Montpellier, Sahlgrenska Academy at the University of Gothenburg, VU University Medical Center, University of Ulm, University of Newcastle upon Tyne, University of Aveiro, University of Szeged, University of Perugia, Ruhr-University Bochum, Heinrich Heine University of Duesseldorf, University of Eastern Finland Kuopio, Institut de la Santé et de la Recherche Médicale, University of Erlangen) as well as four small to medium enterprises. (Matrix Advanced Solutions Germany GmbH, MicroDiscovery GmbH, Protagen, BioGenes GmbH).
cNEUPRO integrates almost all different levels of biomarker research: the primary phase involves the comprehensive clinical characterization of patients and standardized sample-acquisition and handling by specialized geriatric psychiatrists and neurologists. These samples are subsequently used in the search for candidate biomarkers, their biochemical identification by mass spectrometry, and their reassessment in a second, independent set of high quality samples. Finally, the identified biomarkers will be integrated into novel prototype assays (Figure 1).
The research within cNEURPO concentrates on individuals diagnosed with MCI at baseline who subsequently either developed AD, other dementias, or who did not progress to dementia. As the samples had been taken at baseline, clinical information obtained during follow-up allows the identification of predictive biomarker candidates retrospectively. In addition, clinical samples from patients with early AD at baseline or other dementias in the early stages are also included in the analysis.
In the search for new biomarker candidates in CSF or blood, hypothesis-free proteomic approaches such as urea-based gel electrophoresis, Multidimensional liquid chromatography, combined with two-dimensional differential gel electrophoresis (2D-DIGE), several mass spectrometric methods (e.g., SELDI-TOF, MALDI-TOF, nanoLC-MALDI-TOF/TOF, nanoLC-ESI, nanoLCQFTMS), and array-based methods are conducted. Additionally, specific and potentially interesting molecules are studied in detail in the sense of "hypothesis-driven approaches". The most promising biomarker candidates will be selected with the aid of biostatistical tools. Where applicable, published information in terms of the biological function or a possible role of selected candidates in the pathophysiology of AD will also be considered. The selected candidates will be reassessed with a further independent high quality clinical sample of age- and sex-matched patients and controls and with assays allowing for intermediate sample throughput and quantitative comparisons. For those biomarker candidates that can be successfully validated, cNEUPRO will devise novel poly- and monoclonal antibodies. Finally the biomarkers will be integrated into novel ELISA-type assays and, if appropriate, in Multiplex-Assays.
An essential prerequisite for a successful multicenter biomarker-discovery study is the standardization of the clinical diagnostics, the preanalytical sample handling procedures, and the measurements of the known biomarkers total-tau, phospho-tau, and A in CSF. To this end, two neurochemical dementia diagnosis reference centers in Hungary and Portugal are currently being established, and European standard operating procedures for clinical diagnostics and preanalytical sample handling have been defined.
3. Current State and First Results of cNEUPRO
3.1. Neurochemical Dementia Diagnosis-Reference Center in Hungary Launched
In Hungary, 42 Dementia Centers are responsible for the diagnosis and treatment of demented patients. Before 2009, the CSF analysis of A, total-tau, and phospho-tau to support dementia diagnostics was not possible for these centers. As one of the aims of cNEUPRO, the first reference center for neurochemical dementia diagnosis in Hungary was launched in Szeged. With the support of the cNEUPRO consortium, state-of-the-art diagnostic and methodological standards have been implemented, and the center takes part in an ongoing quality control program organized by Kaj Blennow from Sahlgrenska University Hospital, Mölndal, Sweden. During its first twelve months of operation, the neurochemical dementia diagnosis reference center in Szeged has received a total of 54 CSF samples from 14 different Dementia Centers in Hungary. This neurochemical dementia diagnosis center will now try to provide its service to further Dementia Centers in Hungary and to start collecting samples for scientific purposes.
3.2. Diagnostic and Preanalytical Standard Operating Procedures
Due to substantial intercenter variations, the reported accuracy of CSF biomarkers is considerably lower in multicenter studies than in single center surveys [22–24]. To this end, a multicenter study, supported by cNEUPRO, provides guidance on how to establish, validate, and audit CSF tau cutoff values using an unbiased, two-stage multicentre strategy [25]. Furthermore, a hands-on workshop was organized by members of the cNEUPRO consortium (paper submitted to the same issue of IJAD). The aim of the workshop was to assess the differences in assay procedures as potential sources of error. During this workshop, 14 groups simultaneously performed the A, total-tau, and phospho-tau assays according to the guidelines of the manufacturer. At least 23 items in assay procedures were identified that varied between the laboratories, including procedures for washing, pipetting, incubation, finishing, and sample handling. Thus, even if centers use the same assays for A, total-tau, and phospho-tau measurement on a regular basis, they do not uniformly adhere to the procedures recommended by the manufacturer. The results of the workshop stress the importance of standardization of assay protocols. To facilitate biomarker research on a multicenter level, standard operating procedures for the clinical diagnosis and the preanalytical sample handling have been defined by the cNEUPRO consortium (Boxes 2 and 3). The standard operating procedures for sample acquisition, handling, and storage defined by cNEUPRO meet the quality standards required for proteomic studies in CSF [19] and are in agreement with the recently published guidelines for CSF collection and biobanking from the BioMS-eu network [26].
3.3. Investigated CSF Biomarker Candidates for AD Related to Amyloid Precursor Protein (APP) Processing and Tau Pathology
In the last decade, the levels of A peptides and tau proteins in CSF have gained increasing importance in supporting the clinical diagnosis of AD [10, 33]. As no single marker alone allows for a diagnosis with the desired accuracy, several combinations of CSF-biomarkers (A, A, total-tau, phospho-tau) have been proposed [12]. For these markers, a diagnostic accuracy of up to 94% has been achieved in single center studies [12]. Within cNEUPRO, Welge et al. reported a sensitivity and specificity of 88% in the discrimination of AD subjects from other dementias and from elderly depressed individuals with cognitive complaints, by combining the measurement of A, A, and phospho-tau [34]. With the use of MALDI-TOF mass-spectrometry for the study of CSF samples from AD patients, an oxidized form of A (A) was identified. Quantification by SDS-PAGE/western immunoblot revealed elevated A levels in patients with AD as compared to probable vascular dementia and controls [35]. Taken together, these pilot studies suggest that besides A, additional variants of A peptides may turn out to be specifically altered in AD patients.
Although combinations of these CSF biomarkers were reported to have a high predictive value in single-center studies, their application in multicenter-studies is hampered by relatively high intercenter variations. In an associated multicenter study, including 750 patients with MCI who were followed for at least two years, the conversion to AD could be predicted with a sensitivity of 83% and a specificity of 72% by the ratio of A/phospho-tau and total-tau. These values are substantially lower than those seen in several single center studies [24]. The highest intercenter variations were reported for A. As this is probably due to its high potential to form aggregates and to stick to test tubes, alternative markers related to APP processing have been investigated within cNEUPRO. In an associated multicenter study, sAPP and sAPP, two proteins secreted in the CSF after the - or -secretase cleavage of APP, were assessed in 188 patients with MCI or mild to moderate AD. In previous studies, sAPP and sAPP were found to be unchanged [36, 37] or decreased [38–40] in the CSF of AD patients. Within cNEUPRO, sAPP and sAPP levels in CSF of MCI and AD patients with elevated total-tau and reduced A CSF concentrations were compared to those from patients without a respective CSF biomarker profile. Both were found to be higher in the CSF from patients with an AD-indicative biomarker profile [41]. Taken together, these results suggest that sAPP and sAPP may be indicators of altered APP expression and/or metabolism. Reports on their value as candidate biomarkers are however so far contradictory.
In a different study which was supported by cNEUPRO, six novel N-terminal APP-fragments with molecular masses of approximately 12 kDa and starting at amino acid 18 of the APP sequence were detected in CSF by mass spectrometry. In a subsequent small pilot study, six of six AD patients and five of five controls could be classified correctly by the combined evaluation of five of the six fragments [42]. Additionally, Immuno-MS analysis of CSF has led to the detection of eleven novel APP fragments, which begin N-terminally to the -secretase cleavage site, and end one amino acid before the proposed -secretase cleavage site (APP/A peptides) [43]. Interestingly, seven of the twelve APP/A peptides were significantly upregulated in AD [43].
3.4. CSF-Biomarker Candidates for AD Investigated within cNEUPRO, Which Are Not Related to APP Processing or Tau Pathology
One of several kinases that have been suggested to be involved in the abnormal hyperphosphorylation of tau is the MAP-kinase ERK1/2. In a methodological pilot study, ERK 1/2 and its doubly phosphorylated, activated form have been detected in a small number of CSF samples from patients with AD, MCI, and frontotemporal lobar degeneration (FTLD) [44]. To evaluate the usefulness of ERK 1/2 as a potential novel CSF biomarker, ERK1/2 levels in CSF are currently being studied in a total of 110 CSF samples from partners within the consortium with a chemiluminescent 96 well assay format.
In accordance with a previous report [45], research within cNEUPRO found glial fibrillary acidic protein (GFAP), a marker for astrogliosis, to be increased in CSF of AD and sporadic Creutzfeldt-Jacob Disease (sCJD) patients. CSF samples of 18 AD patients, 22 sCJD cases, and 18 from nondemented controls were analyzed with the use of a commercially available ELISA. In AD, a remarkable elevation in CSF GFAP levels with no overlap to controls was observed. Although a significant increase in GFAP could be observed in CJD as well, this was not as pronounced as in AD [46]. Consequently GFAP might have some additive value as part of a biomarker supported diagnosis, although it lacks specificity for AD.
Chronic inflammation associated with oxidative and nitrosative stress is another aspect which is considered to be important in the pathophysiology of AD [47]. The most common protein markers of oxidative and nitrosative stress are protein-bound carbonyls and 3-nitrotyrosine [48]. An increased oxidation of certain proteins and an increased concentration of 3-nitrotyrosine have been reported in tissue [49] and CSF [50–52] of AD patients, but there is also contradictory data indicating no difference between AD and controls [53]. In a study conducted by members of the cNEUPRO consortium, where the concentrations of 3-nitrotyrosine and total protein carbonylation were measured, no change was found in CSF of AD patients [48]. Yet, slightly reduced levels of protein carbonyls were detected in ApoE-ε4 carriers as compared to ApoE-ε4 noncarriers [48]. These results suggest that the concentrations of total protein carbonyls and 3-nitrotyrosine are at this stage not suitable to monitor the chronic inflammatory processes related to AD.
3.5. Investigated CSF Biomarkers for Other Neurodegenerative Diseases
In addition to promoting the early and predictive diagnosis of AD, cNEUPRO is also dedicated to search for new biomarkers to support the diagnosis of other neurodegenerative diseases such as sCJD, FTLD, vascular dementia (VaD), Dementia with Lewy bodies (DLB), Parkinson's Disease (PD), and Parkinson's Disease Dementia (PDD).
Two-dimensional differential gel electrophoresis (2D-DIGE) followed by MALDI-TOF mass-spectrometry indicated that CSF from patients with sCJD differed from CSF from patients with other neurological deficits on the basis of several protein spots. Among these, several previously identified surrogate markers of sCJD such as 14-3-3 protein, neuron-specific enolase, and lactate dehydrogenase were identified. Additionally, an unidentified protein of 85 kDa was found to be significantly increased in sCJD patients [54].
In a separate cNEUPRO investigation, SELDI-TOF mass spectrometry was applied in the analysis of CSF from 32 sCJD patients, 32 controls, and 31 patients with other dementias. Ubiquitin, an 8.6 kDa protein involved in protein degradation, was found to be elevated in the CSF of sCJD cases. This could be confirmed by reassessment with western immunoblots. In the study population, the accuracy of a biomarker-based classification of the samples could be significantly improved by including Ubiquitin in addition to tau, and 14-3-3 protein [55]. This finding is in accordance with several previous reports where Ubiquitin was also found to be elevated in the CSF of sCJD patients [56]. As there is also evidence for altered levels of CSF Ubiquitin in AD [57–59] and vascular dementia [60], it seems that this observation is related to neurodegenerative processes in general and not to a specific disease. Yet, in the Steinacker study CSF Ubiquitin levels in sCJD were higher than those in other dementias [55]. Therefore, Ubiquitin may still be a good biomarker for sCJD if, as with tau protein [61], disease-specific cut-off values are applied.
S100B, another astroglial marker, may also be useful to support the diagnosis of sCJD. Within cNEUPRO, S100B was measured in 54 CSF samples from patients with sCJD, AD, and control patients with the use of a commercial ELISA. Supporting previous findings [62, 63], S100B was shown to be highly elevated in sCJD with no overlap to the other groups [46]. Others have found elevated S100B in familial CJD cases [64], but also in CSF [65] and serum [66] of AD patients. These findings suggest that more attention might be paid to the use of astroglial markers in supporting the differential diagnosis of dementias [46].
With respect to FTLD, cNEUPRO found elevated mean levels of the TAR DNA-binding Protein 43 (TDP-43) and reduced A levels [67, 68]. In line with the reported increased gene expression of TDP-43 in brain tissues [69], elevated 45 kDa TDP-43 levels were found in the CSF of 12 patients with FTLD as compared to 13 nondemented controls by western-immunoblot [67].
In the same sample, the assessment of different A peptide species, sAPP and sAPP, by electrochemiluminescence-based multiplex assays indicated no significant difference for sAPP and sAPP between the groups. However, reduced A levels were found in FTLD [68]. These findings are supported by several earlier studies which found CSF-levels of A in FTLD to be lower than in nondemented controls and higher than in AD [70–73]. However, there are also contradictory publications, regarding levels of Aβ species which did not find reduced CSF A concentrations in FTLD [74, 75]. Although TDP-43 and fragments of APP processing are currently not suitable as biomarkers because of a large overlap between the different diagnostic groups, these findings may still reflect aspects relevant for understanding the pathophysiology of these disorders.
In an associated study focussed on the biomarker supported differential diagnosis of AD, PD, PDD, and DLB, CSF A, total-tau, and phospho-tau were measured in the CSF of a total of 80 patients. Although some significant differences in the average biomarker measurements were found between the groups, only AD patients could be effectively differentiated from patients with other dementias by phospho-tau. For A, total-tau, and phospho-tau, a large overlap between the other neurodegenerative diseases was observed. Interestingly, only in DLB were A and total-tau found to correlate with the duration and the severity of dementia [76]. Consequently, more and better biological markers are needed to support the differential diagnosis of these dementias [77].
A marker with a potential specificity for synucleinopathies may be the lysosomal hydrolase -glucocerebrosidase. In addition to a previous report linking a reduced activity of -glucocerebrosidase to PD [78], a reduced activity of -glucocerebrosidase was specifically found in DLB within cNEUPRO. In CSF from nondemented controls, patients with AD or FTLD, no differences in -glucocerebrosidase activity were found. In contrast, the activity of -mannosidase, another lysosomal hydrolase, was found to be significantly reduced in all investigated neurodegenerative diseases as compared to controls [79]. In order to support the hypothesis that CSF -glucocerebrosidase activity might be a novel CSF biomarker of synucleinopathies, the data need to be confirmed in larger studies.
3.6. Investigated Blood-Biomarker Candidates Related to APP Processing
Several recent studies aimed at identifying AD biomarkers in blood were specifically targeted at determination of Aβ peptides in blood plasma or serum [20].
Within a cNEUPRO associated substudy of the German Kompetenznetz Demenzen (http://www.kompetenznetz-demenzen.de/), A and A were assessed in blood plasma from 257 individuals with multiplexing technology on the Luminex platform. A statistically significant decrease of the A ratio was found in the plasma of the patients with early AD and MCI of AD type whose clinical diagnoses were backed up by corresponding findings in the CSF [80]. Moreover, the cNEUPRO associated French "Three-City study" found that a reduction of the ratios A/A as well as A/A was associated with an increased risk of developing dementia within the next two years [81]. In contrast, several other published studies have not reported significant differences in A peptide concentrations in blood plasma between AD patients and controls [82–84]. In summary, there is no definitive conclusion as to whether plasma Aβ reflects the changing level of central amyloid [20]. Due to the substantial interindividual variations and a large overlap between the diagnostic groups, measuring the individual concentrations of A peptides in plasma is not suitable to support the clinical diagnosis of different dementia disorders. However, there is preliminary evidence that specific forms of A peptides in plasma prove to be helpful in the differential diagnosis of AD and other dementias. In a retrospective pilot study which was supported by cNEUPRO, vascular dementia could be differentiated with a sensitivity and specificity of >80% from other dementias and depressive controls by the ratio of A/A [85].
Currently, highly sensitive assays for the detection of Aβ peptides in blood and CSF are available for A, A, and A. For a detailed analysis of additional variants of Aβ peptides in blood plasma, a highly sensitive two-dimensional gel separation method was established within cNEUPRO. Using this method, at least 30 different A peptides were observed [86]. Semiquantitative analysis revealed that the peptides A and A accounted for less than 60% of all Aβ peptides that were detected by the specific antibody that was used in this study. At least 10% of the detected A peptides appear to be N-terminally truncated [86]. One possible source of these N-terminally truncated Aβ peptides detected in human plasma is mononuclear phagocytes. Cultures of human mononuclear phagocytes were shown to secrete complex A peptide patterns characterized by a high proportion of N-terminally truncated variants [87]. Furthermore, the secretion of A peptides from human mononuclear phagocytes was differentially regulated in response to cell culture conditions [87] and was elevated in cell cultures of mononuclear phagocytes from AD patients as compared to controls [88]. Additional work is under way to evaluate several N- and C-terminally truncated A peptides in plasma as potential biomarkers for AD.
3.7. Currently Ongoing Research in cNEUPRO
The identification of valid biomarkers in blood is highly desirable because they have the advantage of being easily accessible. The search for potential biomarker candidates in plasma or serum is complicated by the presence of a number of highly abundant proteins. These proteins which are believed to have only small diagnostic potential make up about 90% of the whole plasma proteome [89]. As a first step towards biomarker discovery in serum, it was shown that the depletion of 12 high abundant serum proteins by immuno affinity chromatography columns resulted in an increased number of detected peaks by subsequent analysis with SELDI-TOF mass spectrometry [90]. In contrast, CSF proteomics for biomarker discovery in neurodegenerative diseases is particularly attractive because of the proximity of CSF to the brain. Again, the removal of highly abundant proteins resulted in an improved detection of low abundant CSF proteins including brain-derived proteins. Additional separation procedures were introduced to account for the large dynamic range of the expression levels and to simplify the analysis of proteolytically generated peptides by mass spectrometry. For a comparative analysis of individual clinical samples and for a relatively in-depth search for potential novel biomarkers, reproducibility is an absolute requirement. Therefore, different multiaffinity depletion methods followed by gel-nanoLC-MS/MS and spectral counting have been evaluated for the in-depth, label-free quantitative analysis of CSF. Depletion in spin-filter format, coupled to gel-LC-MS/MS, provided a robust method that yielded 800 CSF proteins per analyzed sample, with acceptable reproducibility of protein identification (71%–74% in technical replicates) and quantification (17%–18% CV on spectral counts). To control for reproducibility, the same workflow was implemented in two separate laboratories within cNEUPRO. This proteomics approach was subsequently applied in both laboratories to the independent analysis of two separate cohorts of 20 individual CSF samples each. In both cohorts the patients were clinically diagnosed, and CSF was taken according to the cNEUPRO standard operating procedures. Both discovery sets of samples included CSF samples from five control subjects, from five subjects with mild cognitive impairment without conversion to AD, from five patients with mild cognitive impairment with conversion to AD within the follow-up of 2 years, and five patients with AD. Both datasets contained 1100 identified proteins with a total of 1600 unique CSF proteins in the common dataset and an overlap of 500 between the two laboratories. The biostatistical analysis is currently on-going to select the most promising candidates for a reassessment by targeted mass spectrometry and antibody-based methods in a larger set of samples.
4. Conclusion
Within the first two years, cNEUPRO confirmed sAPP, various Aβ peptide variants, GFAP, S100B, and ubiquitin as biomarker candidates known from previous studies. Additionally, further APP fragments were discovered and TDP-43 as well as β-glucocerebrosidase and ERK 1/2 were proposed as potential novel candidate biomarkers for the early and differential diagnosis of neurodegenerative diseases (Table 1). Because of the high complexity of the blood proteome and probably because of its distance from brain pathology, novel biomarkers in serum or plasma are still elusive. To promote biomarkers in support of the clinical diagnosis of neuropsychiatric disorders in Europe, cNEUPRO devised European standard operating procedures for preanalytical sample handling and established a neurochemical dementia diagnosis reference center in Hungary. cNEUPRO has now started to select the most promising biomarker candidates from two proteomic studies within cNEUPRO and to reassess the most promising biomarker candidates with larger sample size and independent methods to finally integrate them into novel prototype assays.
To increase the accuracy of a biomarker-based diagnosis, biomarkers in body-fluids have been combined with other biological markers such as structural and functional neuroimaging and neuropsychological testing [91]. Whether the new biomarker assays which will be developed within cNEUPRO will be useful in such a multimodal diagnostic workup remains to be elucidated.
The authors were supported by the cNEUPRO consortium (Stephane Roche, Sylvain Lehmann, Ann Brinkmalm, Nicklas Mattson, David J. Burn, Martin Wiesenfeldt, Edgar da Cruz e Silva, Odete da Cruz e Silva, Arif Malik, Johannes Schuchardt, Carsten Korth, Andreas Müller-Schiffmann, Tuula Pirtilla, Stephan Müllner, Angelika Lüking, Johannes Kornhuber). Financial support came from the European Comission (cNEUPRO, LSHM CT-2007-037950, NeuroTAS, LSHB-CT-2006-037953, Anteprion) and the Landesstiftung Baden Württemberg.
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text:
Why Should I Post on Facebook?
Well the idea is to create ongoing massive engagement so that you turn your fans into profitable super fans. These are the ones who don't just engage and tell people about you, they buy from you. The whole ethos for Facebook business marketing is to turn posts into profits.
What Should I Post on Facebook?
Well the more you engage, the more you get seen so you should think of simple at a glance status updates of around 150 characters.
You should also remember that the audience to whom you are effectively selling may not have the same interests as you so step outside your niche, especially if you want them to share your content. When they share stuff, their friends see it, so you want to make people want to share.
Many people put up motivational posts and quotes that will resonate with their audience.
Dan Zarella has statistics which say that Photos get more likes and shares than Posts or Videos or Links but Posts can beat Photos for comments.
The thing about getting likes is that you have to remember to tell people what you want them to do. So, ask for the like in your status update 'Click Like if you like this quote/photo - it's one of my faves'. If you don't give them instructions, then they will just look and then move on.
'I'm putting together a *, let me know if you want a copy by putting yes in the comments box and liking this update' This is a double whammy which can create massive engagement.
And, don't just tell people, create an image to spark their attention. Images in the newsfeed get seen and shared more often than anything - even video - because they are eye-catching and thought provoking. People like them enough to want to pass them on.
A lot of people put up images that include great quotes. Statistics say that the greatest proportion of our learning occurs visually - only 10% will remember what they hear, 20% what they read, whilst 80% will remember what they see.
The images you should be putting up should be:
thought provoking
eye catching
tell a story
In terms of content, easy to answer questions always work well too - fill in blanks, what's the best *, that type of thing.
And don't underestimate the power of contests, giveaways and promos - but remember you cannot do these directly on your Wall. Third party apps need to be used to run them through one of the custom apps under the cover image. You can promote the contest on your wall but you cannot actually host it there, along with a lot of other rules and regulations that can get your Page closed down if you are not careful.
Should I Use Promoted Posts?
Promoted Posts are available to those businesses who have more than 400 likes. They attract the perfect audience and the experts say they are worth paying a bit of extra money to get more reach.
A Promoted Post is a status update on your wall that you pay to promote in the newsfeed to ensure it gets greater exposure to both your fans and the friends of your fans who have engaged with your post. You can only promote a post in the first 3 days it goes live. Amy Porterfield gave an example of one of her own posts which achieved an additional 7,000 likes to a non-promoted version at the cost of $20.
It's about collecting leads and creating relationships with people didnt have access to before. The goal is to reach more of your existing audience and then their fans too.
Sponsored stories are now social ads that allow you to get onto the newsfeeds as well as the right hand column with traditional Facebook ads. They are one of the most powerful opportunities on Facebook by allowing you to reach the friends of fans. Rather than just targeting anonymous users with ads you are much more likely to get a greater response from someone one degree removed who sees that their friend has liked something.
Amy Porterfield's Facebook Mistakes #2 - Killing consistency
If you want to be successful, you need to be consistent about the amount of times that you post. Current thinking is that the sweet spot is 2-5 posts a day with the main reason for unliking a Page being too much posting. So check your insights and see what is working for you.
Amy Porterfield's Facebook Mistakes #8 Lack of Strategy for Timeline
This is your chance to showcase new products, highlight successes for yourself and your clients. Tell people what is going on. Use links in your status updates to send them to your custom apps where you are promoting events or content that contains an opt in feature.
Having said that, you also need to remember what it is that you want to achieve. One of the biggest Facebook user Mistakes is not having a clear strategy. Ask yourself about your core marketing focus and what action you want your fans to take. Are you looking to get more sales by driving them to your custom apps for your products? Or more traffic by sending them to your custom apps which have opt in pages where you can gather their email address in return for a future discount or promotion? Or brand awareness with your changing cover image of your products and status updates about furture launches and events.
Make sure that your content is interspersed with links to your custom apps or pages on your website that will provide interest.
Amy Porterfield's Facebook Mistakes #10 Over Automation
Everyone can tell when content has been autoposted so get on there yourself and interact regularly. You can improve your engagement score if you get into conversations with your status updates. You can also increase likes and shares by your fans if you make sure that the content you share has an eye-catching picture and an engaging headline.
Amy Porterfield's Facebook Mistakes #11 Ignoring Insights
Your insights are the stats that tell you what content is working. If you get good ratings, do some more of the same. But don't be tempted to compare your Insights with another Page - you don't know if they are running an ad or sponsored story to drive traffic to their page.

subset: commoncrawl


text:
A woman has been charged with the shooting murder of a man at an Adelaide car wash on the weekend.
The 35-year-old is charged with murdering 39-year-old Adrian Paul MacDonald from Hope Valley on February 3, police said on Tuesday.
He was shot at about 6.40am (CDT) at the back of the Big Bucket Car Wash at Parafield.
The woman was arrested late on Monday at an Elizabeth Park home.
The woman and the victim were known to each other, police said.
They are calling for public help from anyone who saw a 2003 silver Ford Futura sedan in the Parafield area or neighbouring suburbs on Sunday morning.
The car has been seized by police and is being currently being forensically examined.
The woman is expected to appear in Adelaide Magistrates Court later on Tuesday.
Detective-Inspector Mark Trenwith told reporters the seized car was owned by the arrested woman's family and was at the carwash at the time of the murder.
"The post-mortem yesterday confirmed the cause of death was multiple gunshot wounds," he said, adding that the weapon had not been recovered.
"We do not believe the murder is related to drugs or any other illicit (activity)."
He said police are continuing to view CCTV footage from the area.
"We believe that will form an important part of our case in regard to the arrest of this woman," Insp Trenwith said.
Police could not rule out the possibility of further arrests.


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South Dakota student to lead national CTSO
Carter Christensen, of Sioux Falls, was elected national president during last months national DECA convention. Christensen, a Lincoln High School senior, will serve full-time for the next year as the organizations student leader.
The career and technical student organization has approximately 200,000 high school and collegiate members worldwide and prepares students for careers in business, marketing and financing. Like all CTSOs, DECA places a big emphasis on community involvement.
Click here to watch a KELOLAND interview with Christensen.


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A lot of people create alter-egos on the infamous Twitter for the same reason people like LARPing or putting on trenchcoats and flashing little old ladies. It's just something to do. My good friend and colleague Ian Healy decided it might be fun if we pretended to be jaded and (eternally drunk) editors at an unnamed publishing house. We called the account @FakeEditor, and with the flourish of a red blingy pen, we sent out our first tweet on April 22nd, 2010.
|There's no better Rickroll than this in existence
It took a little time to pick up attention, but it was no less fun as we traded back and forth our own little barbs about the industry behind our cynical little guise. Eventually we started gaining steam through holding contests and networking with a lot of authors. @FakeEditor attended the Pikes Peak Writers Conference in 2011 and did good bit of Tweeting, causing a stir by making several people wonder if they were sharing the table with the infamous curmudgeonly drunkard. Eventually we added the "Wise Ass Intern," who would send out Tweets on Wednesday, thereby increasing the multiple personality disorder between the two of us. At various points, we threatened to out ourselves, or we implicated other Twitter personalities, such as @EvilWylie to throw people off our scent. We even decided to post a picture of ourselves, which went over really well with followers.
Over the course of my time under the alias, I passed a couple Tweets with Stephen King's son Joe Hill. At some point, (after my departure), Wil Wheaton even re-tweeted @FakeEditor, and I think by then, Ian and I both agreed that his little idea had finally reached legend status.
I eventually lost the ability to maintain dual personalities, so Ian kept on with it, bringing on the "intern" permanently to share the margaritas and fake drudgery. We have decided not to out this person, but we figured it was time to unmask ourselves. No real reason other than I guess we both agreed it was time. Why the hell not, you know? I'm sure the shenanigans will continue over at @FakeEditor. After two years, the account currently has over 4500 followers, which is several times more than the number of followers Ian and I have combined (and we've both been on Twitter twice as long). It's odd, really. @FakeEditor never interacts much with other users, and we never had a policy to serial follow people or do any of the other things that "experts" say guarantees more followers. Everyone involved just made a point to be as funny and interesting as possible, and I guess that worked out. I think it's all about being raw, unfettered personality.
At any rate, even though I am no longer involved, I highly recommend (especially if you're a writer or editor or agent) to give @FakeEditor a follow. Ian and his current intern (who is as yet unnamed) are damn funny. I might even pop on from time to time to say something, but the beauty is, you'll never really know for sure.


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I really wanted to keep things fairly simple. Last year we constructed felt Valentine's envelopes for all the kids (even the two on the way) and I had stuffed a few trinkets and surprises inside for our Valentine's breakfast. It was fun, but I felt like maybe Valentine's Day would start to resemble a smaller version of Christmas stockings. This year I decided to ditch the gifts and focus on card-making and sweets.
I prefer living in a small town, but really, REALLY miss the convenience of Target, Michael's, and Party City when there's creative work to be done or a party to plan. My life doesn't allow much time for trips into Madison, so with a little help from Pinterest and the Baraboo Wal-Mart, our Valentine's wheels were put into motion.
|Valentine production: Day One. Watercolors, aluminum foil, and stickers...William decorating his cards.
|Valentine production: Day Two. I failed to snap an in-progress photo with six painty hands sitting at the table, but the little boys did hands (and baby feet) so they'd have cards to pass out, too.
|Next, plenty of sweets and treats! I found these homemade gumdrops on Pinterest and thought they were cute. William and I made them together and packaged some for friends.
|Sugar cookies with Ahmie! Mmmm...best cookies ever.
|Charlie was so proud. What a big shot.
|Mailboxes and envelopes galore...6 adults, 6 kids.
|Here come the troops...William's valentines ready to distribute, Benny and Ikey reading Valentine books while we wait.
|Filling envelopes!
|Charlie Bear
|Benny Boo
|Hanky and John John...my "twins." Betsy made them the most adorable matching heart onesies!
|Just like brothers!
Hope you had a love-filled day!
|Happy Valentine's Day! With love from us...


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Moving On
It has been a little over two weeks since I was laid off and I have been fairly buzzing with energy. Relatively speaking, I'm doing great. I think it helps that right from the start I sized up the situation pragmatically, considered my options, made priorities and a game plan, focused on my successes and didn't take getting laid off personally. I don't mean to sound like Pollyanna or anything but seriously, by not sitting down and letting waves of desolation wash over me and think, "ah, woe is me!" but instead, maintaining an active state of "do something, and do it now!" I have been able to ride this dip in the road fairly well.
I'm lucky, I don't have any debt other than a mortgage, which I just refinanced at a spectacular 3% for 15 years (yeah, that pretty much rocks) and fortunately was able to finesse that a month before I was laid off. I set about trimming even more fat off my budget and by living frugally I can live on severance and unemployment for a significant amount of time. I did some research and a strong candidate, in this market, can usually be assured of finding adequate employment in 3-6 months. My goal is by the end of July. I'm doing the job search full time, networking, using LinkedIn, other social media, job boards, referrals. I will be employed and expect to employed well.
I've even got a spin, a truthful one, on being laid off. As my former employer put it, it was a business decision. As a software sales executive, I covered NY, NJ, New England, and Canada, and any open territory in the rest of the United States. I did this all remotely from Western Pennsylvania. We did not have the budget to send me to meet clients face to face, go to networking events or conferences. This was a terrible handicap vs. our competitors. But now, the company is in the process of merging with a small software company in NYC that has 3 sales people in my former territory. One of those sales people will be taking over my territory.
And it makes sense. I'd make that decision if I was running the company. So no looking back and on to new opportunities, bigger and better ventures.
I've also turned my attention to my house and all those projects you never have time for when you are juggling a professional life, personal life, family, friends, and everything else that comes along. My garage has been completely organized and cleaned out, even had a company come and haul out the computer monitor, two televisions, printer, scanner, VHS player, window air-conditioner and other sundry items taking up half of my garage.
I also pulled out boxes of old files and went through filing cabinets and found documents, even taxes, credit card receipts, stuff dating back to 1986! Boxed it all up and took it off to be shredded by a professional, bonded company in town. Done!
Another priority I made was to focus on physical fitness. This is something I started over a year ago but, let's face it, with no job to go to every day, you can really make time for some major workouts every day.
My girl friend, who is a local caterer, and I, meet three times a week to walk/run 4 miles and do 74 steps up a steep ridge where we live. We do them twice. I'm also doing Les Mills Pump (a barbell with weights-based workout) several times a week.
Diane, the caterer, and I just worked out an agreement, where I will provide sales/marketing services in exchange for physical fitness training a few times a week. A win-win situation so I can put something on my resume during my time "off".
So what's my point of telling you all of this? I know this post is a bit different than what I've written before but so many people have been in this situation, especially with this rollercoaster economy. Being unemployed can be one of the most devastating things to happen to someone, especially when you have a family to support. But I adamantly believe (and have written this before) that you define a situation by how you respond to it. Action brings results. Positive thinking, about yourself, what you have to offer and what the future will bring, opens the door to new opportunities.
So raising a glass to all of you that are in the same boat, good luck to you and give it your all. Persevere! I do think that things are looking up. I see more activity on job boards like The Ladders, Careerbuilder, Monster. But make sure you use LinkedIn and your professional and personal network.
This is also a great time to explore something you've always wanted to do, something you feel passionate about. I know once I get some of these projects completed that I am looking forward to spending more time writing and cultivating my creative Muse. This happened for a reason and I am embracing the moment.


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Harmony Clinic Has a Bilingual Doctor
Did you know that Dr. Lester Carrero of Harmony Clinic speaks both Spanish and English?
Serving the local community is a priority for Harmony Clinic, and being able to communicate in Spanish and English is incredibly valuable.
Dr. Carrero, originally from Puerto Rico, is proud to serve the Latino community for all chiropractic and wellness needs.
Additionally, our website can be switched from English to Spanish by selecting one of the flags in the top right corner.
When you need a chiropractor, don't hesitate to call us at (386) 774-7246 for an appointment! Harmony Clinic is conveniently located in Orange City, FL, close to nearby communities such as Deland, Deltona, DeBary, Enterprise, and Lake Helen.
2 Comments - Leave a Comment


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About Our Trust
Established in 1972, the Mount Dora Community Trust is a tax-exempt charitable organization created to provide the local community with funding for public educational, charitable or benevolent projects and purposes.
The Trust continues the mission to provide outreach programs to schools, charities and civic causes through the generous actions and financial support of Mount Dora residents and businesses committed to promoting the community's unity and prosperity.
Would you like to be a part of this important cause? We would greatly appreciate your support! To find out how to help secure a bright and vibrant future for Mount Dora, please click here.
What is the Mount Dora Community Trust? A more accurate question might be "Who?" because the Trust is composed of citizens whose collective goal is to safeguard and advance the community through support for local education, youth sports and activities, and benevolent causes assisting those in need.
These civic-minded individuals and businesses have united to assure a thriving future for our community by performing the following functions:
The First National Bank of Mount Dora oversees the administration and investment of the Trust's funds. John Pease IV, Senior Vice President and Trust Investment Officer, oversees the investments with Elizabeth "Susie" Pelfrey as the Portfolio Manager; C.E. (Ed) Brooks III is the Senior Trust Advisor at the First National Bank of Mount Dora.
- A City of Mount Dora Council-member – our current councilperson is Laurie Tillett; previous members have included, Vasco Watters, former Mayor, Melissa DeMarco, Glenna Burch, Judy Smathers.
- A member of the Board of the Chamber of the Commerce – our present member is Kim Stewart; past representatives have included Betty Hensinger, Barbra Perfecto, Lou Biron, and William Brooks, III.
- A member of the Ministerial Association of Mount Dora – currently serving is Pastor Tom Biery; past members have included Pastor Tom Baker, Chaplain Alan Jones, Reverend Woodleigh Volland, and Jim Keough.
- A rotating seat among the local service clubs (Rotary, Lions and Kiwanis) – this seat is now represented by John Stewart of the Kiwanis Club of Mount Dora; previous seats have been held by James (Jim) Croson of the Rotary Club, Mary Pezzo of the Lions Club, and Glenna Osborne of Kiwanis.
- A member of The First National Bank of Mount Dora's Board of Directors – our present Director is Sam Sadler III, who replaced "Mr. Mount Dora" himself, Dick Edgerton.
The Trust helps newer organizations, such as, Hoops Scholarship Fund,fulfill their respective missions while providing continuous funding to longtime successes such as The Center for the Arts, The Library Association, and the Mount Dora Area Chamber of Commerce.
Wherever you look in Mount Dora, whether it's children's programming, community health issues, artistic projects or educational opportunities, the Mount Dora Community Trust has been a strong and vigilant partner.
How You Can Help
For our partnership with the community to grow even stronger, we need your help. Please remember us in your charitable contributions and your estate planning. We are happy to work with you and/or your attorney, accountant or financial advisor.
One of our previous members said it best: "There is no greater satisfaction than doing something for someone you will never know, and no greater reward than seeing the benefits of your generosity take hold."
With questions about how you can help promote better education, youth programs and assistance to our community in Mount Dora, please contact Grant and Scholarship Manager Kevin J. Batliner at firstname.lastname@example.com.
Thank you for your generous support.


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The Book Blogosphere is getting bigger and bigger every day – it has gotten so big in the last ten years that I find it hard to navigate sometimes. And some of the hardest places to navigate are actually getting those books to review.
Getting Started
The ARC and review copy are the backbone of our blogs. If we don't have books to read, we can't review them and to stay with trends, it's always good to read the new books. Getting new books means we are in search of ARCs and review copies. You can go about this in a few different ways. If you want to read a scolding post about the "proper" way to blog for ARCs or a holier-than-though post about the care and feeding of ARCs, this isn't it. If you are in search of ARCs for nefarious purposes, my two cents will probably not sway you, so I'm just going to lay out the facts.
- The first WAY to get an ARC is by requesting them. To do this you need to get an email address for the publisher and the RIGHT one. These are usually handled via interns or Marketing Assistants, which tend to change a lot, but usually, each publisher has a general email address and that general email address will be forwarded to the person handling the marketing for that particular book. To get the email address do a google search "Publisher Name, Request Review Copies." Searches like this will lead you to pages like this. Know the book you want, know the publisher and then request. Follow the rules for each imprint. Write a nice email (channel professionalism, think job interview or query letter) and request your book. Quick tip – put the Book Title and ISBN number in the subject with the words REVIEW REQUEST. (See email list below)
- Contact the Author directly. This doesn't work all the time, usually an author has only a few ARCs of their book and if an author is with a traditional publishing house, they'll have PR people handling their reviews. Authors that are with boutique presses, indie and are self-published will handle their own ARCs, so you'll have better luck with contacting these authors.
- Online Outlets. In today's market, bloggers are being directed to online sites like NetGalley and Edelweiss, it is probably your most likely chance to get an ARC of a book. These will be digital galleys. (See List Below)
- Conventions are an amazing way to get ARCs. If you can afford to attend a convention like ALA, BEA, RT … to name a few, you'll have your choice of ARCs. The publishers have printed ARCs for the sole purpose of handing them out to media, bloggers, librarians and publishing professionals that attend these conventions.
FYI – I specifically did not include groups that trade ARCs, you can get ARCs this way, but I have not used them in the past and only recommend the sites that get permission from the Author and the Publisher. If you haven't gotten permission to "trade" or lend out your ARC, the publisher might not like this and prevent you from getting ARCs in the future. Since I personally have not vetted any of these sites, I thought it would be prudent to not include them. This does not mean they are not legit, just that I can not certify them. Thanks for understanding. Now that you are ready…check out the list below and have fun. I look forward to reading your reviews!
Where to get eARCs and Review Copies?Online Sites & Requests Via Publishers
Direct Request Contact Pages:
- Penguin Random House
Media Queries Page
- Hachette
Imprint Emails Page
- HarperCollin
Publicity Contact Pages
- Macmillan
Publicity Contacts
- Simon & Schuster
Media Emails Page by Imprint
- Scholastic
Media Contacts
- Houghton Mifflin Harcourt
Media Contacts
- Egmont
Press Office Email: firstname.lastname@example.com
Publisher Sites
Sometimes the publishers will have reader/blogger sites that offer ARCs. Most of the time it works best if you direct request from the publisher, but a lot of bloggers have gotten ARCs and review copies from the groups.
- Random House Reader Circle
- Penguin Teen for Bloggers
- BookNook (Hay House)
- First to Read (Penguin)
Christian Publishers
- B&H Blogger Partnership Program
- BookLook Bloggers (HarperCollins Christian Publishing)
- Crossway Blog Review Program
- BookLook Bloggers (HarperCollins Christian Publishing)
Professional Sites
- Blogging for Books
- Book Browse
- Edelweiss
- Goodreads First Reads
(Giveaway Program)
- LibraryThing's Early Reviewer's Program
- NetGalley
- The Reading Room
- Shelf Awareness Pro
(subscribe to the newsletter and they will offer ARCs in their content and via the ads on the sidebar)
- Story Cartel (Indie)
Know of any other resources for ARCs and Review Copies?
5 Comments
nordie@writing about books on September 29, 2016 at 08:58
Develop a relationship with authors, e.g. by going to events and actually talking to them. I've got a number of authors who now contact me directly to ask me to review their books ahead of the launch
Jennifer @ The Book Nympho on September 27, 2016 at 12:13
These days I'm more of an audiobook requester.
Carmel @ Rabid reads on September 27, 2016 at 11:12
Thanks for the list! I recently discovered SocialBookCo. They have titles for review, but you can also add up to 5 already published books on your "wishlist", and if selected, they will send one to you. There's also Manic Readers for indie titles, and Audiobook Jukebox for well, audiobooks, although their selection isn't that good.


text: So, was there some kind of defective copy of Sonic Adventure 2 Battle sold at any point? Cause I've had the game for a while and there have been glitches, but yesterday about four or five times it kept giving me the An Error has Occurred message and that's never happened before now. I don't want my GCN to be failing like my PS2 did. I've never had any Nintendo systems crap out on me... Well at least not without my being the catalyst. I think my SNES stopped working right because of the half a dozen times I yanked the cartridge out without shutting the game off beforehand. And my original Gameboy had one to many accidents falling out of my hand so some of the game reading chips broke off. None of my NES systems ever developed the won't play games at all problems though as far as I can recall. I don't see any disk blemishes or scratches, but it's hard to tell on those tiny disks especially if the scratch is like less then an inch long. Any insight would be helpful. But considering SA2 is the only game I've ever encountered problems with I have to assume it's gotta be the disk itself and not my system.


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Immediately After an Assault
What Should I do Now?
Make sure you are safe from further harm. Call someone you trust, such as a friend, your CA/RA, the Office of the Vice President of Student Affairs, the Rape Response, and/or the University Police Department.
Do not bathe or douche. Do not eat or have anything to drink. Try not to urinate if possible.
If oral contact took place, do not smoke, eat, or brush your teeth.
Go to the hospital emergency room. Medical attention is vital, as you may have injuries of which you are unaware. They can also test you for sexually transmitted diseases, and provide emergency contraception if desired.
If you've already changed clothes, place them in a paper bag (plastic destroys evidence). If you haven't changed, keep the original clothes on, and bring an extra set to wear home from the hospital. The police may need to keep your clothes for evidence.
"Working Together for a Safer Community"


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The Kouga Municipality Disaster Management team has called off the search for the possible site where a meteorite may have crashed.
The search was launched in the area around Patensie earlier on Monday morning with the police and a team from the Baviaans Nature Reserve following several reports from people who saw a bright flash and minutes later a loud bang.
Kouga Municipality spokesperson, Mfundo Sobele.
"We are hoping that someone might come across something and point us in the right direction, but for now the search is off" he said.
Source: Algoa FM


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Policies and Practices
The Registrar's office is the custodian of the Institution's academic and curricular records.
The links below are the institutional rules and regulations for some of the policies pertaining to maintaining academic record.
For information regarding institution Academic Rules and Regulations, see below:
Specific Policies for students:
- Academic Plan Hold (Requires an undergraduate student to declare a valid major)
- Family Education Rights & Privacy Act (Regarding your rights about your academic record)
- UW Extension Independent Learning Tuition Waiver (Fee Waiver policy)
- Students Exceeding 165 Total Credits (Fee policy)
- Legal or Preferred Identity Practices (Name, Gender, Social Security Number)
Did You Know? Credit Hour Definition: A credit hour is an amount of work represented in intended student learning outcomes and verified by evidence of student achievement that is an institutionally established equivalency that reasonably approximates not less than one hour of classroom or direct faculty instruction and a minimum of two hours of out-of-class student work each week for approximately fifteen weeks for one semester, or the equivalent amount of work over different amount of time, or the equivalent amount of work for other activities established by the University including but not limited to graduate work, internships, practica, studio work, and other academic work leading toward the awarding of credit hours.


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To the Congress of the United States:
In accordance with the Impoundment Control Act of 1974, I herewith report five revised deferrals of budget authority now totaling 9,663,811.
The deferrals affect programs in the Departments of Agriculture, Defense-Civil, Energy, Health and Human Services-Social Security Administration, and Justice.
The details of the deferrals are contained in the enclosed report.
George Bush
The White House,
April 18, 1989.
Note: The attachments detailing the deferrals were printed in the ``Federal Register'' of April 27.


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I have planned a visit to Italy during Christmas for 4 days. But I want to cover number of places.
I will be landing in Pisa on 22-Dec-06 in Evening and will be coming back on 26-Dec-06 from Milan.
I have planned to visit Pisa, Florance, Rome, Venice, Pompai and Milan.
But not able to arrange these in optimal order. Can u help me in planning my visit??? like which order I shd visit the places so that I visit all the places and also I shd nt miss nice places in each city.....
Also the safe budgeted accomodation.....
Any suggestion like dropping a city and adding new place is most welcome.


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Maybe you mean: 'Homepage' or 'camera-installation-2' or 'home_automation' or 'work_in_action' or 'low-voltage-integrated'
It's easy to raise or dim any light in the room-or entire house-right from a touch screen.
Elegant and Efficient Lighting
Smart lighting is one of the biggest opportunities for you to subtly, but effectively, influence the ambiance of your home and complement the aesthetics of its decor. You can turn on the entire room-or multiple rooms-with a single light switch, and ramp and dim illumination to whatever level you desire. And if you don't care for the banks of switches on your wall, your lighting controls can be centralized elsewhere, such as in a closet, to avoid distracting from the style of your home. Smart lighting is both beautiful and energy efficient. Your lighting automatically turns on when you need the light, but turns itself off when no one is in the room.
- Have the lights in the house turn on automatically to welcome you home.
- Use motion sensors to provide hands-free illumination.
- Program the lights to shut off automatically when a room is unoccupied.
- Use keypads to trigger more than just lights, including music, shade control, and energy settings-with a touch of a button.
- Illuminate the hallway lights automatically for safe entry, as soon as you enter the garage.
- Have your outdoor lighting turn off in the morning, and turn on as the sun goes down.
- Make your home sophisticated enough to appear to be occupied even when you're away.
Configurable keypads are personalized for your home and can create any mood or ambiance with one touch.
The Control4 Wireless Thermostat has more than 50 years of climate control experience inside, and is built specifically for home automation.
Smart, Comfortable and Convenient
Being comfortable encompasses more than just adjusting the temperature. Control humidity levels, automate shades, power on the hot tub on your way home, and program your entire house for maximum energy efficiency. Control4 works with the world's leading brands, so it's easy to integrate your automation system with radiant flooring, forced air, dual fuel and geothermal systems.complete the experience.
- Wireless thermostats maintain consistent temperature and humidity control, regardless of the time of day or season.
- Enjoy personalized comfort settings that can be recalled on a schedule or with just a few taps on your phone or touch screen.
- Pool and spa controls make it easy to turn on the heat, set the temperature or crank the jets instantly, right from your smartphone.
- Fireplace sensors are easily controlled from any device within the home. Press the customized "Romance" button and enjoy.
- Program your window shades to lower during the hottest hours of the day, saving on air conditioning costs.
- Automate your sprinklers and lawn systems to respond to weather conditions.
Personalized comfort settings can be recalled with just a few taps on your phone.
Enjoy fast, full-screen picture and high-quality audio from every touch screen.
High-Definition Video Intercom
Whether you want to see who is at the front door, check in on your sleeping baby or call everyone to the table for dinner, it's simple and convenient with video intercom. With an incredibly fast connection speed and high-definition clarity, all of this functionality is automatically enabled n every touch screen in a Control4 Smart Home. on your smartphone, touch screen or TV.
- Replace your doorbell with an elegant Door Station to see who is at the door without getting off the couch.
- Get a complete view of the front porch with alternate camera views.
- Call any touch screen within your home, whether to check in on the kids or bring everyone together
Door Stations let you see who's at the front door, and multiple cameras provide complete views of the porch.


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Wings Over Willcox 2014
This popular annual event offers birders the opportunity to see more than 25,000 wintering Sandhill Cranes, along with hawks, eagles, falcons, and more than twenty species of sparrows. The program includes bird tours with expert guides, seminars and a nature expo with plenty of vendors and live animals. There will also be tours and seminars on Sierra Vista's history, geology, natural history and more. For more information visit www.wingsoverwillcox.com/
Date: 15-19 January 2014
Venue: Sierra Vista
State: Arizona
Country: United States
Create a Safe Haven for Birds in Your Garden
November 20, 2012 by Editor
Filed under Birding Tips
As urban areas become more and more built up, birds and wildlife are increasingly being forced out of their natural habitat. Gardeners can do much to alleviate the plight of birds by putting some thought into planning a garden that will make their feathered friends feel at home. This need not be complicated, and certainly need not be costly, as even a small bird-friendly spot in an urban garden can be a life-sustaining oasis that will more than reward the gardener for his, or her, efforts.
In planning a bird-friendly garden there are a number of points to bear in mind, one of the most important being to provide a clean source of water in a spot where birds have an easy escape route should they be disturbed by a predator, such as the neighborhood tabby. Recent statistics published by the RSPB noted that it is conservatively estimated that cats in the United Kingdom catch up to 55 million birds each year, with the most frequent victims being house sparrows, blue tits, blackbirds and starlings. A birdbath on a stand, placed beneath a tree, allows birds a view of their surroundings while they splash in the water, while giving them a quick escape route into overhanging branches should they feel threatened. Spiny and prickly plants such as holly can be grown around and beneath feeders and bird baths to discouraged cats from lurking there. Ensure that the bird bath is free of algae and filled with fresh water.
When choosing what to plant in your bird-friendly garden, it is best to go indigenous, as this will provide local birds with what they need, when they need it. Of course, birds are very adaptable and will make use of exotic plants as well, if it suits their needs. A good garden center in your area should be able to advise you on what to plant to attract birds, and a mix of indigenous and exotic plants can work well. Give consideration to including plants to provide food such as seeds, nuts, fruit and berries in all seasons, nesting materials and shelter. Food can always be supplemented with strategically placed feeders and nesting boxes are often welcome.
It should go without saying that pesticides should never be used in a bird-friendly garden. Given enough time, nature will take care of its own pests, and until ecological balance is reached, gardeners may need to put some extra effort into controlling pests organically and removing them by hand. Birds are great at keeping creepy-crawlies in check, so invite them into your garden and enjoy their company, while they enjoy the safe haven you have provided.