Alzheimer's disease (AD) is a progressive, multifactorial neurodegenerative disorder that is the main cause of dementia globally. AD is associated with increased oxidative stress, resulting from imbalance in production and clearance of reactive oxygen species (ROS). ROS can damage DNA and other macromolecules, leading to genome instability and disrupted cellular functions. Base excision repair (BER) plays a major role in repairing oxidative DNA lesions. Here, we compared the expression of BER components APE1, OGG1, PARP1 and Polβ in blood and postmortem brain tissue from patients with AD, mild cognitive impairment (MCI) and healthy controls (HC).
BER mRNA levels were correlated to clinical signs and cerebrospinal fluid biomarkers for AD. Notably, the expression of BER genes was higher in brain tissue than in blood samples. Polβ mRNA and protein levels were significantly higher in the cerebellum than in the other brain regions, more so in AD patients than in HC. Blood mRNA levels of OGG1 was low and PARP1 high in MCI and AD.
These findings reflect the oxidative stress-generating energy-consumption in the brain and the importance of BER in repairing these damage events. The data suggest that alteration in BER gene expression is an event preceding AD. The results link DNA repair in brain and blood to the etiology of AD at the molecular level and can potentially serve in establishing novel biomarkers, particularly in the AD prodromal phase.
Alzheimer's disease (AD) is a progressive, multifactorial neurodegenerative disease affecting 24 million individuals worldwide, with an incidence expected to double within 2030 . AD is thus by far the most important contributor to cognitive decline and dementia globally. AD is characterized by impairment in memory and cognition, synaptic dysfunction, neuronal loss, extracellular amyloid beta (Aβ) plaques and intracellular neurofibrillary tangles (NFTs) composed of fibrillar aggregates of hyperphosphorylated tau in the brain . Low Aβ-42 in cerebrospinal fluid (CSF) may reflect deposition of amyloid in brain plaques, while high total tau (T-tau) and phosphorylated tau (P-tau) in CSF may reflect neuronal degeneration [3, 4]. These CSF biomarkers are used in the clinic for diagnosis of patients with AD [2, 5, 6]. However, it remains a challenge to diagnose early stages of multifactorial sporadic (non-familial) AD. This is consistent with the hypothesis of multiple underlying factors leading to sporadic AD, whose etiology remains poorly understood.
Age is the principal risk factor for AD and ageing is associated with cumulative oxidative stress. A leading hypothesis proposes that high levels of oxidized nucleic acids in brain cells can lead to neuronal dysfunction in patients with AD [7–9], and the evidence linking oxidative damage to neurodegeneration is overwhelming [10–12]. Consistent with this, reactive oxygen species (ROS) can damage macromolecules such as lipids, proteins, DNA and RNA. At the same time, it is clear that genome dynamics and defects in DNA repair processes under the oxidative stress induced in neurodegeneration hold a key to main understanding in neuropathogenesis. Because environmental genotoxic components generally fail to pass through the blood–brain barrier, it is thought that endogenous ROS are the primary cause of oxidative DNA damage inside neurons and glial cells [13, 14].
DNA damage is balanced with repair in a homeostatic process, and imbalance occurs when the damage exceeds repair, causing cellular senescence, genome mutation or apoptosis. These features are more abundant in old cells than in young cells [15, 16]. A high level of DNA damage can be particularly deleterious in post-mitotic cells as they do not self-renew through cell proliferation. However, oxidative damage to DNA and RNA may be a cause or consequence of neurodegeneration, reflecting either increased production of ROS or reduced DNA repair . Previous studies show that the amount and the capacity to repair DNA damage varies with age and varies in different brain regions, and that pathological features of AD are higher in brain regions where more DNA damage is detected [14, 17, 18]. BER is the major pathway for repair of oxidative DNA damage  and epidemiology studies have associated reduced BER capacity with neurodegenerative diseases such as AD [20, 21]. The first step in BER usually involves excision of a damaged base by a lesion-specific DNA glycosylase, e.g. when OGG1 removes 8oxoG from an 8oxoG:C base pair , which generates an abasic (apurinic/apyrimidinic; AP) site . In the next step in BER, AP-endonuclease 1 (APE1) hydrolytically cleaves the phosphodiester backbone at the AP site, after which polymerase beta (Polβ) performs end-processing and gap-filling DNA synthesis, and finally, DNA ligase seals the nick . Polβ is the major DNA polymerase and tissue-specific expression levels of Polβ has been reported . Poly (ADP-ribose) polymerase (PARP1) plays a role in many DNA repair reactions, including BER [25–27].
In AD, brain pathology can be observed many years before cognitive decline is clinically evident. CSF biomarkers indicate brain pathology compatible with mild cognitive impairment (MCI) preceding AD , and central nervous system amyloid deposition can be determined either by CSF Aβ42 or amyloid PET scans in preclinical AD . At the same time, AD brain pathology may be present in the absence of cognitive impairment. However, inexpensive, non-invasive biomarkers for pre-dementia are lacking, and additional biomarkers to identify individuals at risk for dementia and other types of cognitive disease/decline are non-existent.
We have previously shown that there are alterations in mRNA levels of APE1 and OGG1 in tissue from different brain regions of the tg-ArcSwe mouse model and that these occurred prior to the development of AD pathology . As mouse models only represent models of imposed AD pathology and do not reflect human AD in complexity, we asked if BER mRNA levels were altered between human AD patients and healthy controls (HC) in blood and in brain tissue. The present study explores the relationships between the quantitative analysis of mRNA transcripts for the BER genes encoding APE1, Polβ, OGG1 and PARP1 and CSF biomarkers of AD and clinical signs of cognitive decline in blood from a clinical cohort (n = 166) as well as in a second cohort of postmortem samples of different brain parts (hippocampus, cerebellum, entorhinal cortex and frontal cortex) from AD patients (n =42) and HC (n = 9). The results show that alterations in the BER gene expression profile in blood is an early event evident already at the prodromal stage of AD. This could be a useful indicator for disease progression/initiation status in patients with preclinical AD.
This cross-sectional study was performed using two separate cohorts. The first cohort included 166 live individuals at different disease stages relevant to AD: 41 AD patients with dementia, 28 patients with MCI due to AD pathology, 45 patients with MCI and 24 patients with subjective cognitive impairment (SCI) and 28 HC recruited at the Memory Clinic at Akershus University Hospital Additional file 1: Table S1). Assessments of these patients included lumbar puncture (LP) for CSF sample, blood draw, brain MRIs (except in cases where CT scan was preferred over MRI), formal cognitive testing including mini mental status evaluation (MMSE) , Cognistat , geriatric depression testing and neurological examination. For further information on diagnostic eligibility criteria and patient groups, see the Additional file 1.
The second cohort included freshly frozen postmortem human brain samples of the hippocampus, entorhinal cortex, frontal cortex and cerebellum obtained from 42 histopathologically confirmed AD patients (average age 82.4, age range 57–98 years, 20 males/22 females) and 9 human controls displaying no AD histopathology at time of death (average age 61.7, age range 46–84 years, 5 males/4 females, for more patient characteristics see Additional file 1: Table S2). The brain samples were provided by Fundación Centro Investigación Enfermedades Neurológicas (CIEN)/Carlos III Health Research Institute (ISCIII).
CSF and blood analysis were performed in cohort one. Blood samples were drawn by venipuncture and CSF samples (5 mL) were drawn by LP under spinal anesthesia or during an otherwise scheduled diagnostic procedures. LP was performed between 09–12 AM. The concentration of T-tau, P-tau and Aβ-42 in CSF was measured using protocols developed at Akershus University Hospital (Additional file 1: Tables S3 and S4). APOE was determined using DNA isolated from blood samples, and the APOE allele frequencies are listed in Additional file 1: Table S5.
Blood samples were collected in PAXgene RNA collection tubes (PreAnalytiX GmbH, Switzerland). The samples were stored at −80 °C until use. Total RNA was extracted using the PAXgene Blood RNA kit (PreAnalytiX GmbH, Switzerland) in accordance with the manufacturer's recommendations. Human brain samples were weighed and homogenized using MagNA Lyser Green Beads (Roche Diognostics GmbH, Mannheim, Germany) and lysis buffer from PureLink® RNA Mini Kit (Ambion, Texas, USA) in a MagNA Lyser instrument (Roche Diognostics GmbH, Mannheim, Germany). Total RNA was isolated using PureLink® RNA Mini Kit according to manufacturer's recommendations. The RNA concentration of both blood and brain tissue was determined using an ND-1000 spectrophotometer (NanoDrop technologies, Saveen & Werner AB, Sweden); RNA purity, integrity and yield were confirmed using Agilent 2001 Bioanalyzer and RNA 6000 Nano Kit (Agilent technologies, California, USA) according to the recommendations of the manufacturer. RNA samples with unsatisfactory purity (blood RIN < 7, brain RIN < 5) were isolated again until satisfactory purity and RIN was obtained or dismissed. cDNA was prepared from 1000 ng total RNA from blood and brain tissue in 100 μL using the High cDNA reverse transcription kit (Applied Biosystems) according to the recommendations from the manufacturer (Invitrogen, US).
mRNA transcripts encoding OGG1, APE1, Polβ and PARP1 were quantified by real time q-PCR (qRT-PCR) using methodology and equipment by Applied Biosystems (Foster City, CA, USA). Complete TaqMan gene expression assay information is listed in Additional file 1: Table S6 and relative mRNA levels (mean, range) for all groups are listed in Additional file 1: Table S7. To control for differences in efficiency of the reverse transcription and real-time PCR reactions and pipetting errors, normalization to a reference gene was also included (in addition to other normalization procedures, please see Additional file 1 for further information). The selection of the reference gene was performed after a validation of 32 candidate reference genes using the TaqMan Human Endogenous Control Plates (Applied Biosystems, Foster City, CA, USA) according to the recommendations of the manufacturer. Glyceraldehyde phosphate dehydrogenase (GAPDH) was identified as one of the most stable reference genes with the additional preferred Ct value (<25) and was thus chosen as the endogenous reference gene. qRT-PCR was performed by using the StepOnePlus™ system in 96 well plates with TaqMan Gene expression assays according to the recommendations of the manufacturer. Samples were held at 95 °C for 10 min, cycling was at 95 °C for 15 min and 58 °C for 1 hour for 55 cycles. The melting curve cycle was at 95 °C for 15 min, 60 °C for 1 hour and 95 °C for 15 min. All plates were set up according to the relative standard curve method with separate standard curves on each plates for both target and reference gene. All standard curves were made from the same sample, thus also functioning as a positive control and reference sample between runs. The relative gene expression of target genes was calculated using the relative standard curve method; briefly, this is the difference in Ct value and y-intercept between the target gene and a calibrator sample, divided by the slope and normalized to the reference gene (GAPDH) and adjusted for minute efficiency (see Additional file 1 for more analysis details). Deep sequencing of cDNA from a subset of brain samples was performed in parallel using the Illumina HiSeq-2000 sequencer (50 base pairs single reads, mean read depth 22379938,5 (SD 167932,5), BGI, China). Fragments per kilobase Million (FPKM)) of APE1, OGG1, Polβ and PARP1 values of the individual samples are listed in Additional file 1: Table S8.
Peptide characterization and quantitation were performed by electrospray-based high resolution mass spectrometry (Q-Exactive, Thermo-Fischer). Brain tissue was lysed in lysis solvent containing 2%SDS/10 mM Tris–HCl, pH7.5 supplemented with protease inhibitor cocktail (EDTA free, Roche) and PhosStop (Roche) and disrupted with MagNa Lyser instrument (Roche) in cycles until fully lysed. Prior to trypsin digestion, 100 μg of protein lysates were precipitated over night with acetone at −20 °C. Air dried protein pellet was re-suspended in 10 μl of 8 M urea. Proteins were reduced with 1 μl of 10 mM DTT (Sigma-Aldrich) followed by alkylation with 1 μl 50 mM iodoacetamide (Sigma-Aldrich). The samples were then diluted with 50 mM ammonium bicarbonate and digested with 1:100 of trypsin (sequencing grade modified, Promega, USA). The digested samples were fractionated by anion exchange column and samples were run on a Q-Exactive (Thermo Scientific, Germany) mass spectrometer coupled directly to an nLC (EASY 1000, Thermo Scientific, Germany) using a data-dependent Top10 method. Mass spectrometry results were searched using MaxQuant software against the human UniProt database with proteome ID:UP000005640. For further methodological specifications, please see the Supplementary material (Additional file 1: Table S9.).
To test for differential expression of BER genes among AD and non-AD individuals in the brain samples, a linear mixed model was fitted to each gene in each brain region. The fixed effects were disease status (AD or HC), brain region (hippocampus, cerebellum, frontal cortex or entorhinal cortex) and the interaction between disease status and brain region. A random individual effect was included to account for the potential correlation between measurements in different regions within the same brain. The significance of the overall interaction was assessed by an F-test. If the overall interaction was not significant, the effect of disease status was assessed directly with a t-test (Table 1). If the interaction effect was significant, each brain region was analyzed separately by fitting a reduced model. It included the fixed effect of brain region, interaction between disease status and brain region, and random effects. For each brain region, a t-test was used to test whether the interaction of disease status and brain region was significantly different from zero (Table 2). A significance level of 0.05 was used for this analysis. For each clinical covariate (diagnosis and CSF biomarkers) and each gene, a linear regression model was fitted with the gene expression as the dependent variable. APOE was adjusted for by including it in the model together with the clinical covariate. If the clinical covariate was categorical, each level of the covariate was compared to a reference level. The significance of the clinical covariates was assessed with a Wald test (Table 3). A two-sample t-test was used to test for significant differences in gene expression between brain and blood and between different brain parts (Table 4) Due to the large number of tests in the last mentioned three analysis (Tables 2, 3 and 4), the results in these tables should be interpreted with care. In this setting, it is not clear how one should correct for multiple testing, as many of the tests are dependent. Applying e.g. the Bonferroni criterion would be too strict. However, many of the results were significant even when correcting the p-values with the Bonferroni correction, and are thus reliable. The Pearson correlation was calculated between the RNA deep sequencing data and the RT-qPCR gene expression data.
In order to investigate the relationship between BER mRNA levels in blood and brain tissue, we compared the gene expression from the different brain regions with gene expression in blood in AD patients and HC. The mRNA levels for APE1, OGG1, Polβ and PARP1 was in general significantly higher in brain tissue compared to blood (Fig 1, Table 4). The difference was greater in AD patients for all four BER components in general, showing significant differences also when comparing blood to each brain region for all components and all brain regions under study (Table 4). There was also a significant difference between mRNA levels in blood and brain tissue in HC for APE1, OGG1 and PARP1 in general. When comparing blood to each brain region, significant differences were found for APE1, OGG1 and Polβ in cerebellum only (Table 4). These findings are also evidence of the intact integrity of the postmortem brain tissue mRNA and high BER transcriptional level compared to blood.
Quantitative real time PCR was used to assess mRNA levels in tissue from four different brain regions from 42 AD patients and 9 HC. APE1 mRNA was significantly lower in the entorhinal cortex of AD patients (p <0.05) than in the same cortical area in healthy controls (Fig. 2a, Table 1). Polβ mRNA was significantly higher in the cerebellum of AD patients (p <0.00001) than in the same brain region in healthy controls (Fig. 2b, Table 1) and PARP1 mRNA was significantly lower in AD cerebellum (p < 0.0005) compared to HC cerebellum and significantly higher in AD hippocampus (p < 0.005) and entorhinal cortex (p < 0.0001) compared to the same regions in HC (Fig. 2c, Table 1). OGG1 mRNA levels did not differ between AD and HC brain regions.
Polβ mRNA was higher in cerebellum compared to all other brain regions in both AD and HC. Particularly, Polβ mRNA was significantly different between cerebellum and hippocampus (p < 0.00005) and entorhinal cortex (p <0.001) and frontal cortex (p <0.01) within AD patients and between cerebellum and hippocampus (p <0.01), entorhinal cortex (p <0.005) and frontal cortex (p <0.005) within HC (Fig. 2b, Table 2). Expression of OGG1 and PARP1 did not differ between AD patients and HC in general or in any specific brain region.
The progression in the BER repair mRNA profile in the different phases of AD progression was investigated. We compared the gene expression of APE1, OGG1, Polβ and PARP1 in 41 AD patient with dementia, 28 patients with MCI due to AD pathology, 45 patients with MCI and 24 patients with SCI and 28 HC. OGG1 mRNA was lower in AD dementia (p <0.05), MCI/AD (p <0.05) and patients with MCI (p <0.01) than in HC (Fig 3a and Table 3). PARP1 mRNA was higher in patients with AD dementia (p <0.05), MCI/AD (p <0.01), MCI (p <0.01), and SCI (p <0.01) than in HC (Fig. 3b and Table 3). Polβ mRNA was higher in patients with AD dementia (p <0.05) compared to the other groups (Fig. 3c, Table 3). The abundance of the APE1 mRNA levels did not correlate significantly with any clinical diagnostic marker in any cohort subgroup.
The neuropathological hallmarks of AD are Aβ plaques and NFT and the biomarkers currently used to assess these features are the levels of Aβ-42 and tau in CSF. To explore the relations between BER and the pathological features of AD, we compared the BER mRNA profile in blood in individuals exhibiting 1) only high Aβ-42 (n = 19), 2) only low P-tau or high T-tau or both (referred to as abnormal tau from here on) (n = 24), 3) both high Aβ-42 and abnormal tau (n = 29) levels in CSF exceeding cut-off levels (see Additional file 1: Table S3), and 4) patients with normal levels of CSF Aβ-42 and tau (n = 64) and 5) HC (n = 28) displaying normal CSF levels of CSF Aβ-42 and tau. Thus, we have grouped the individuals according to biological correlates based on their CSF levels of Aβ-42 and tau and not by clinical disease status. For information on which patients are presented in each biological group, see Additional file 1: Table S4. OGG1 mRNA was lower, in the patients with only high Aβ-42 levels (p <0.05), only abnormal tau levels (p <0.05) and patients with normal CSF levels of Aβ-42 and tau (p <0.05) than in HC. There were no significant differences in OGG1 mRNA between patients with both abnormal Aβ-42 and tau levels and HC (Fig. 4a and Table 3). PARP1 mRNA was significantly higher in all patients groups than in HC: e.g. high Aβ-42 levels (p <0.01), abnormal tau levels (p <0.01) and combined high Aβ-42 and abnormal tau (p <0.01), as well as patients with normal CSF levels of Aβ-42 and tau (p <0.0001) (Fig. 4b and Table 3). The abundance of APE1 and Polβ mRNA was not correlated with any of the CSF biomarkers.
Former studies suggest that DNA repair capacity is altered with age so we wanted to explore how the BER mRNA profile changed over time. PARP1 mRNA was negatively correlated with age (p <0.05) with an average effect per year of −0.009 (Table 3), while Polβ mRNA was positively correlated (p <0.01) with age with an average effect per year of 0.010 (Table 3). APE1 and OGG1 mRNA levels were not correlated with age.
When comparing results from RNA deep sequencing and RT-qPCR there was a strong correlation between the expression of Polβ in both methods (95.7 % (p = 0.00000009), with particularly high expression of Polβ in the cerebellum samples. There was no correlation for APE1 (6.3 % (p =0.83), OGG1 (−1.8 % (p = 0.95) or PARP1 (−20.6 % (p =0.48).
Protein detection by next-generation mass spectrometry (MS) demonstrated relatively high protein levels of APE1 and PARP1 in the cerebellum of both AD and HC, while Polβ was only detected in AD cerebellum, however, at a lower level than APE1 and PARP1 (Fig. 5 and Additional file 1: Figure S1). APE1 was reduced in the frontal cortex of HC and totally absent in frontal cortex of AD patients, indicating a reduction of APE1 protein levels in the frontal cortex in general that is more pronounced in AD. PARP1 remained high in the frontal cortex of HC, but was reduced in the AD frontal cortex. OGG1 was not detected by MS in any of the samples.
The goal of this study was to investigate the levels of mRNAs and protein profiles of selected BER enzymes in brain tissue and blood, potentially as early markers of AD development. Notably, mRNA levels of APE1, OGG1, Polβ and PARP1 were higher in brain tissue than in blood, reflecting the high energy consumption of the brain. Brain cells have a high metabolic rate with high glucose and oxygen turnover, creating substantial amounts of ROS. This oxidative stress, in combination with the post-mitotic state of neurons and a possible decreased ratio of antioxidant to pro-oxidant enzymes , make brain cells prone to be dependent on efficient and active BER repair. Our findings support the importance of BER in DNA repair in the brain, consistent with its high load of oxidative stress and DNA damage.
Notably, Polβ mRNA and protein levels were particularly high in the AD cerebellum compared to all other brain regions. This was also verified by RNA deep sequencing analysis, showing a 95,7 % correlation between the two transcriptomic methods (Additional file 1: Tables S7 and S8). The high level of Polβ may be a sign of late AD pathology, as the cerebellum remains free from tau-pathology until the most advanced Braak stage (VI), and neuronal loss and gliosis in this region has not been associated to early AD. Studies show that mice lacking Polβ have neonatal lethality with abnormal neurogenesis characterized by apoptotic cell death specifically only in the developing central and peripheral nervous system , implicating Polβ as an important factor for the nervous system already at the earliest stages of development. Neurons depend heavily on Polβ in the repair of oxidative DNA damage [34, 35], and single nucleotide gap-filling activity and protein level of Polβ was reduced in AD inferior parietal lobule (IPL) samples compared to HC . Since it has been shown that other BER enzymatic activity, such as base excision, abasic site incision and nick ligation was not altered in brain tissue from AD patients, it has been suggested that Polβ is rate limiting for repair . A study on the 3xTgAD/Polβ mouse model, displaying 50 % reduced Polβ activity, demonstrated significant increase in DNA damage accumulation. The reduced DNA repair capacity triggered neuronal death and hippocampal atrophy that did not occur in the 3xTgAD mouse. Thus, Polβ deficiency in combination with Aβ accumulation may comprise the ability of neurons to support synaptic activity to survive and render neurons vulnerability in reduction of cellular energy levels [36, 37], resulting in dysfunction and death. Growing evidence suggest a connection between AD and aspects of energy metabolism such as impairment in insulin  and insulin-like growth factor (IGF) signaling causing deficits in brain oxygen-glucose utilization and that the insulin resistance causes defects in the detoxification systems for oxidative stress . PET imaging of AD brains demonstrate an AD related cerebral glucose metabolic covariance pattern with decreased metabolism in the temporoparietal regions and relatively increased metabolism in the subcortical white matter, cerebellum and sensorimotor cortex . Thus, cerebellum presenting with high Polβ gene expression and increased metabolism may represent a compensatory mechanism for deficits in other brain regions or merely reflect a generally highly active brain region, potentially relating to the high density of granular cells in the cerebellar cortex . Other studies have demonstrated lower levels of Polβ protein and Polβ activity in the cerebellum of AD patients compared to controls . Our proteomic findings indicate that Polβ protein levels were lower than that of the other BER components and were only detectably increased in the AD cerebellum.
APE1 mRNA was significantly lower in the entorhinal cortex of AD patients than in HC entorhinal cortex. The entorhinal cortex is one of the first regions to be affected in AD  and alterations observed here may represent early events of AD progression. However, the findings observed in post-mortem tissue represent late changes in the progression of AD. One study showed that APE1 proteins levels were similar in IPL and cerebellum tissue from AD and HC , while other studies demonstrated that APE1 protein expression was higher in brain tissue affected by AD pathology (hippocampus and surrounding temporal cortex)  and in cell extracts from AD patients . Our proteomic data show relative high protein levels of APE1 in the cerebellum of both AD and HC, however, this was reduced in the frontal cortex of HC and totally absent in frontal cortex of AD patients, indicating a reduction of APE1 protein level in the frontal cortex in general that is accelerated in AD. APE1 has been shown to play a role in degrading damaged RNA , and oxidatively damaged RNA has been implicated as an important factor in neurodegeneration [46, 47].
In blood, OGG1 mRNA transcript abundance was reduced in MCI, MCI/AD and AD patients compared to HC as well as in patients with abnormal levels of CSF Aβ-42 and tau and in patients with normal CSF levels of Aβ-42 and tau (mainly comprised of MCI and SCI patients). The results thus indicate that BER mRNA profile alterations occur independent of plaque and tau pathology in the progression of AD since the alterations are also seen in patients with no CSF pathology, but not in HC. This is consistent with findings from other studies . OGG1 repairs oxidized guanine, and numerous studies show elevated oxidative lesions in both DNA and RNA in the prodromal phases of AD as well as in AD [18, 49–51]. It is suggested that oxidative DNA damage increases only during the early stages of AD and then decreases with the progression of the disease due to activation of a compensatory mechanism .
Since OGG1 expression level was reduced in all groups except SCI, but also in patients with normal CSF levels of Aβ-42 and tau, we suggest that suppressed or reduced OGG1 function is not directly associated with Aβ or tau pathology, but that lower OGG1 transcript levels may indicate a DNA repair deficit in patients with MCI, MCI/AD and AD. Thus, OGG1 may represent a general marker for DNA repair deficits in subjects prone to develop AD . However, it is important to emphasis that SCI and MCI are heterogeneous conditions that may or may not proceed to AD. Other studies show increased OGG1 mRNA levels in brain tissue from the hippocampus, parahippocampal gyri and middle temporal gyri of patients with preclinical stages of AD compared to HC , suggesting that the elevation represents a compensatory increase in protein expression to moderate loss of activity due to posttranslational modification in response to increased oxidative DNA damage. MCI brain tissue did not exhibit a difference in OGG1 protein level compared to HC, but a significant decrease in OGG1 enzyme activity , proposing an association with increased post translational modification of OGG1 by 4-HNE. We were not able to detect OGG1 protein level by mass spectrometry in our study, suggesting low OGG1 levels in brain tissue from both HC and AD patient frontal cortex and cerebellum. Thus, OGG1 results from various studies are conflicting, and it still remains to be determined if OGG1 mRNA and protein levels correspond to enzyme activity. OGG1 has also been implicated in immune system regulation and inflammation [54, 55], and it is suggested that there is a correlation between the efficiency of the DNA repair system and development of inflammation associated with the production of Th1 cytokines.
PARP1 mRNA levels were higher in blood from all patient groups compared to HC. PARP1 is a general marker of DNA damage and inflammation and may also contribute to plaque formation and neurodegeneration in AD patients. Both plaque formation and neurodegeneration in the brain are associated with inflammation possibly mediated by NF-κB , a master regulator of the response to pro-inflammatory stimuli [56, 57] and cellular senescence . PARP1 is required for NF-κB-dependent gene transcription  and NF-κB-dependent gene expression is associated with aging in mouse and humans . Thus, PARP1 connects inflammation and the DNA damage response (DDR), through which excessive DNA damage can lead to cellular senescence. Senescent cells secrete pro-inflammatory cytokines, feeding a vicious cycle .
PARP1 mRNA levels were also higher in patients with high levels of CSF Aβ-42 and abnormal tau and the two combined. However, PARP1 mRNA levels were also higher in patients with normal CSF levels (comprised mainly by MCI and SCI patients) than in HC, indicating that there are processes relevant for AD development (inflammation, senescence, apoptosis and possibly also DNA repair deficiency) that are independent of tau and Aβ pathology. Thus, PARP1 may be a general early indicator of these other processes in all these groups as these changes are already evident in the SCI stage. In AD frontal cortex tissue, however, PARP1 protein levels were lower than in HC, reflecting late stage disease. One study demonstrated that PARP1 activation causes neuronal death in the hippocampal CA1 region by increasing the expression of Ca2+-permeable AMPA receptors , suggesting that increased PARP1 may cause damage to neurons. PARP1 thus functions at the center of cellular stress responses, where it processes diverse signals and, in response, directs cells to specific fates based on the type and strength of the stress stimulus. Thus, PARP1-stimulated senescence, apoptosis or necrosis could further deplete the pool of regenerative cells, and thereby contribute to neurodegeneration .
Consistent with other studies, we found that PARP1 expression in blood decreased with age , while Polβ increased with age. Total BER capacity has been shown to be inversely correlated with age in healthy controls, but not in AD patients, however, reduced BER associated with AD regardless of age has been suggested to be linked to a premature ageing phenotype .
DNA repair capacity and protein levels differs among ethnic groups and there is considerable inter-individual variation . Some of this enzymatic variation is most likely to be due to post-translational modifications. Our data demonstrated that the integrity of mRNA in post-mortem brain tissue was intact, as false negative results could have been an issue if mRNA levels were lower in brain due to post-mortem degradation. The transcriptomic analysis was reproducible and standardized equally for the two clinical cohorts, controlling for tissue differences and normalized in multiple steps to assure correct analysis, including the validation of GAPDH. Even though cohort differences might explain some of the differences in gene expression observed between blood and brain, the considerably higher level of gene expression measured in the brain than in blood is beyond inconsistent findings resulting from individual variability or from bias in the cohort of post-mortem samples. The discrepancy between transcriptomic and proteomic findings are most likely due to post-transcriptional processes, where high gene transcription levels not always reflected in a similar protein level.
In summary, the data presented here provide novel insight into the early pathophysiology of AD, and OGG1 and PARP1 can potentially contribute as part of a set of blood biomarkers for identifying incipient AD. Early prediction of insipient AD or AD-like pathology is critical to the management of this disease and may also facilitate the design and evaluation of diagnostic, preventive and therapeutic tools for AD and AD-like forms of dementia.
AD, Alzheimer's disease; APE1, AP endonuclease 1; Aβ, Amyloid abeta; BER, Base excision repair; CSF, Cerebrospinal fluid; HC, Healthy controls; MCI, Mild cognitive impairment; NFTs, Neurofibrillary tangles; OGG1, 8-Oxoguanine glycosylase; PARP1, Poly [ADP-ribose] polymerase 1; Polβ, Polymerase β; P-tau, Phosphorylated tau; ROS, Reactive oxygen species SCI, Subjective cognitive impairment; T-tau, Total tau.
This research was supported by the Oslo University Hospital (Rikshospitalet) and Akershus University Hospital. We gratefully acknowledge the contributions of Ramune Grambaite for clinical assessment and lumbar punctures and Berglind Gisladòttir for contribution in assembling clinical parameters on a sub-selection of the patients.
We are thankful for grants from the Research Council of Norway Centre of Excellence (CMBN) project 145977 to TT, project 194056/V50 to TT and TF, and project 2014020 to MSL and TT and and project 2014050 to TT from the South-Eastern Norway Regional Health Authority.
The complete datasets supporting the findings are presented in this manuscript and in the Supplementary material provided.
MSL, TT and TF conceived the study and study design; MSL, TT, MS, TR, DM, YE and LM performed technical analyses; AR collected the biobank of human brain tissue samples; TF, VS, and PS performed the clinical analyses; CCG performed the statistical analyses; MSL, TF and TT wrote the paper; all authors commented and approved the manuscript and consented for publication.
All study protocols were approved by the Norwegian regional ethical committee for medical research (REK 2011/1034 and REK 2013/1643), and written, informed consent was obtained from all study subjects prior to start of clinical tests. The analyses of brain tissue were approved by both the Research Ethical Committee (Research Ethical and Animal Welfare Committee, ISCIII, Spain) and the Norwegian Ethical Committee.


The Minister of Economy, Trade and Tourism Mihai Tudose discussed on Thursday with the Ukrainian Ambassador to Romania Teofil Bauer the ways of boosting the bilateral trade and investments, the Ministry of Economy, Trade and Tourism (MECT) informed in a release.
The two officials "set up a series of joint action to develop the collaboration on all the economic levels and the advance of commercial exchanges; an important role was attributed to the talks within the second session of the Romania-Ukraine Joint Intergovernmental Committee for Economic, Industrial, Technical and Scientific Cooperation in Bucharest, in 2015.
On December 31 2014, the trade between Romania and Ukraine totalled 1,439 dollars, comprising of 826,4 million dollars exports and 612,78 million dollars imports, resulting in a balance surplus of 213,62 million dollars for Romania. The volume of this trade was 24.0 smaller than in 2013 (minus 35.7 percent for exports and plus 0.7 percent for imports). Ukraine is Romania's 5th export market outside the European Union, with 4.07 percent of the total exports. Conversely, 3.07 percent of Romania's non-EU imports came from Ukraine, placing this country 8th among the non-EU trade partners.


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Average 27 yr old Male needs job to support my hobbies.
[KS] Required to take a break?
Job offer for graphic artists.
Now we have to worry about our Facebook score?!
Any vehicle builders in the KC area?
Interview this coming Wednesday and Thursday, last minute advice.
Any Software Engineers on here?
Who has got demoted at work?
School me on the Series 6!?
Breaking into the IT field?
Looking to hire a detailer with experience.
Where to find a good Resume form?
Software people, what do you follow?
CAD question / which software?
Looking to hire someone to detail from the 13th to 25th, paying cash.
Does anyone know anybody in Chicago?
Seeking Advice: What would you do?
Anyone have an Associates in Computer Science?
Need a job for 30 days.
BCS Recruiters - Anyone used them?
Do you have illustration skills / design experience?
READ BEFORE POSTING A "LOOKING FOR A JOB" THREAD!!
has anyone gone to Centriq?
Those of you who don't have a college degree..
Job openings at NTB in olathe!
Looking to hire a detailer! Full-Time, Monday-Friday 9-6.


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Students'protest shrinks in numbers, they demand support from the US and EU embassies
Nga Erdi Ibro Publikuar më: 09-01-2019, 09:37
The students' protest continued on its second day after the holidays, despite the fact that the number of demonstrators in front of the Prime Minister's office had fallen significantly. On the other hand, lecture halls were once again empty as a very small number of students have turned up for classes
The number of students that participated yesterday in a protest held in the capital has shrunk. In contrast to Monday, fewer students gathered in front of the government's offices on Tuesday. They were about 50 students with placards in their hands, repeating their 8 demands for the government, adding that they insist on not engaging in dialogue. During their protest, students have marched toward the US embassy, where they have delivered a letter which was also addressed to the EU. In this letter they asked for support about their 8 demands and also made a ninth demand for the abrogation of the law on Higher Education. "This was a spontaneous decision that we had not predicted. I believe that embassy officials will support us", one of the students said. Protesters in front of the Prime Minister's office said that they will not give up until their demands are met. According to them, the low participation was due to the low temperatures that have swept the country. Prime Minister Rama has reacted by addressing a message to all those skeptic students who still don't believe in the decision taken by the government to cut tuition fees by 50% this academic year for all students and to cut tuition fees by 100% for all of those students who have excellent grades.
"Those who decided to be involved in ugly scenes of brutality and ignorance, are trying to misinform students about the tariffs. Nobody will pay more than 50%", Rama said.
On the other hand, there's also been a reaction by the Ministry of Education following the announcement made by the head of the government, Edi Rama regarding tuition fees for students. The ministry declared that it will allocate the necessary fund which will subsidize the fees that the government will pay for students and scholarships, adding that this institution is working to identify the number of students who benefit from the government's decisions.

redpajama_set_name: RedPajamaCommonCrawl


Jesus said to them, "If God were your Father, you would love me, for I came from God and now I am here. I did not come on my own, but he sent me.
We are inheritors of identity politics. We are just as divided as the Jews were in Jesus' time. The Jews identified as sons of Abraham, yet sectioned off from each other because of ideological differences. The Essenes fled to the desert, the Pharisees prayed within their clan, the Sadducees kept busy in the Temple, and the Zealots plotted to blow the whole thing up. They could not identify with each other. Is this not the current landscape of our world?
Then there's Jesus, who identified himself as a child of God, the Son of God. It's no wonder scholars struggle identifying Jesus' Jewish faction. Perhaps it's because he only identified with God, as God. He knew that identity politics was nothing more than tribalism, colonization, and dominance. It wasn't going to save anyone.
—Mark Chang is a Theology Teacher and the Director of the New Teacher Induction Program at Loyola Academy in Wilmette, IL.
and of the reward of eternity.


503.3 Marking. Where required by the fire code official, approved signs or other approved notices or markings shall be provided for fire apparatus access roads to identify such roads or prohibit the obstruction thereof. Signs, notices or markings shall be maintained in a clean and legible condition at all times and shall be replaced or repaired when necessary to provide adequate visibility. The owner, manager, or person in charge of any new or existing building or property for which fire lanes have been approved or required by the fire code official shall mark and maintain said fire lanes as provided in this subsection.
503.3.1 Curb marking. All curbs and curb ends shall be painted red with white lettering at least three inches high with at least one half-inch stroke, stating "FIRE LANE," and at least one of the following phrases: "NO PARKING," or "TOW-AWAY ZONE". Wording shall not be spaced more than 25 feet apart.
503.3.2.1 Signs. Signs that comply with Appendix D section D103.6 shall be mounted conspicuously along the edge of the fire lane, spaced no more than 25 feet apart, and shall have lettering at least two inches high with three-eighths inch stroke.
503.3.2.2 Stripes. From the point the fire lane begins to the point the fire lane ends, there shall be painted one continuous red stripe having a minimum width of at least six inches. Fire lanes shall be stenciled every 25 feet apart, stating "FIRE LANE," and at least one of the following phrases: "NO PARKING," or "TOW-AWAY ZONE," with white lettering at least three inches high with at least one half-inch stroke. Fire lanes shall be so marked in accordance with Appendix D sections D103.6.1 and D103.6.2.
All designated fire lanes shall be maintained and kept in a state of good repair at all times by the owner or person in control of the premises. Upon notification that the fire lane is not clearly marked, the fire lane shall be repaired or repainted to meet the city code. Failure to maintain the fire lane shall constitute a violation of the fire code.


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The Good Place Season 5 Premiere Date & News: Everything You Need To Know!
Nobody knows what happens after we die, and any guess is as good as the next. The Good Place is a fantasy comedy that explores what happens after we die. We follow Eleanor (Kirsten Bell) as she is transported to The Good Place and laments the fact that she is in "heaven" despite the fact that she does not deserve it.
However, as the series progresses, we gradually realize that things aren't as "good" as they appear. If you haven't seen the series yet, it's available on Netflix, so go check it out.
The Good Place Season 5 Release Date
Season 4 of NBC's 'The Good Place' premiered on September 26, 2019, and concluded on January 30, 2020. There will be 14 episodes instead of 13 for the first time since the first three seasons. On NBC, you can watch all of the episodes.
In June of this year, NBC announced that 'The Good Place' would end after season 4.
Since the show's renewal for a second season, Schur and the writers have been discussing how to proceed.
We've considered making more than four seasons at various points over the last few years. Nonetheless, we decided against it because making this show is such a unique and rewarding creative experience that we don't want to waste it. Our final season will begin in the upcoming fourth.
During a subsequent interview, Schur discussed whether the season finale met his expectations. It was difficult for me to predict where we would end up at the start of this. It would have been extremely arrogant of me to create a pilot and then say, "When this is finished in four years, this will be a fantastic concept." "I believe we started talking about the ending in season three," he said.
"We began to imagine and extrapolate where we were going in a very generic sense," Schur continued. We had a good idea of what would happen after season three.
That was the plan heading into season four, and we stuck to it. The conclusion of the show, like many other plot points, evolved naturally. We improved on the original concept rather than abandoning it."
At this point, the fifth season appears to be doomed. Season 5 of 'The Good Place' has been formally canceled by the show's producers. With the series now available on Netflix, fans can catch up on their favorite episodes whenever they want.
The Good Place Season 5 Plot
After so many reboots and revivals, it's difficult to say goodbye to a series. It's difficult to predict what NBC's web-based feature, Peacock, will offer now that it's so close. The Good Place Season 5 would make the most sense if the original cast and characters remained the main focus.
Season 5 of The Good Place could reveal an escape clause stating that leaving The Good Place does not completely erase one's existence. Return to Michael and Janet in center four for a fascinating explanation.
Team Cockroach may also encounter a problem with the afterlife structure, causing them to return to the beginning. However, for the time being, consider Season 4 of The Good Place to be the final goodbye.
The Good Place Season 5 Cast
If the show returns, all of the original cast members are expected to return to the set.
Eleanor Shellstrop is played by Kristen Bell, Chidi Anagonye is played by William Jackson Harper, Tahani Al-Jamil is played by Jameda Jamil, Jason Mendoza is played by Manny Jacinto (Top Gun Maverick), Janet is played by D'Arcy Carden, and Michael is played by Ted Danson (Curb Your Enthusiasm).
Where To Watch The Good Place
Watch The Good Place | Netflix.
The Good Place Season 5 Trailer
The fifth season of The Good Place has yet to be announced. As a result, the Season 5 trailer has yet to be released.
Let's take a look at the trailer for The Good Place's fourth season.
Babylon Berlin Season 4: Release Date Status Set For 2022 On Netflix
To Your Eternity Season, 2 Release Date Status Announced: All Details Are Here!
The Lego Movie 3 Release Date Status, Cast And Crew, Plot, Trailer And More
Yashahime Season 3 Release Date Status, Expected Cast & Plot, And Trailer Update
Did good place get Cancelled?
"Congratulations to Mike Schur and an outstanding cast and crew for working together to create a thoughtful, mind-bending, and hysterical series unlike anything else on television," they said. "We can't wait to see what surprising stories the new season brings." Season 4 sadly marked the end of The Good Place.
Where is Chidi from The Good Place?
Kaguya-sama: Love Is War Movie U.s. Confirmed Release Date In Trailer


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Home › eBooks › Billie Holiday
Authored by: Forrest Cole
From the Series: Black Americans of Achievement
Her tragic life and death are as important a part of Billie Holiday's story as her undeniable talent as a singer. Poverty-stricken from birth, she was introduced to drugs and hard living at a very young age and spent her childhood on the streets carousing with various criminal elements. But, among that turmoil she found her greatest strength—her unique and emotion-filled voice—and a willingness to succeed. Although her emphatic delivery and natural talent were initially off-putting to the mainstream, her perseverance helped to transform the role of the vocalist in popular music. All singers who have followed, regardless of the genre they sing in, owe a debt of gratitude to Billie Holiday. In this insightful and thorough biography, readers will learn about the troubles faced by this legendary singer and how she affected the music world with her original voice.
Benjamin Hooks
Carol Moseley-Braun
Charles Drew
Henry Louis Gates, Jr.
James Farmer
Mae Jemison, Updated Edition
Muhammad Ali, Updated Edition
Paul Laurence Dunbar
T.D. Jakes
Terry Mcmillan
W.E.B. DuBois


Home God What was so evil about eating a piece of fruit in the garden of Eden?
What was so evil about eating a piece of fruit in the garden of Eden?
God created Adam and Eve with free-will which entailed that they should have a test to prove their loyalty. Not eating from the tree of knowledge of good and evil was God's test to them (Genesis 2:17). Eating the fruit was a sin because it was disobedience to God's clear command. When Adam and Eve disobeyed, they brought upon themselves separation from God, suffering and eternal death (Isaiah 59:2).
But the only One who paid the price of men's disobedience was God Himself. The price of redeeming the human race was too high. God offered His only Son to redeem mankind from eternal death "For God so loved the world that He gave His only begotten Son, that whoever believes in Him should not perish but have everlasting life" (John 3:16).
Thus, through the disobedience of Adam, sin entered this world and through the obedience of Christ, salvation became available to all who accept His free gift (Romans 5:17). Man's condition for remaining in Eden is also the same condition for being restored to Eden. Therefore, the most evidence of a truly converted person is to break the pattern of disobedience.
And those who will be restored to the lost paradise will have to demonstrate that they can be trusted through obedience (1 John 5:3). Their steadfast loyalty will be an eternal guarantee of the security of God's restored kingdom. Therefore, those who look so lightly upon the good works of obedience are in error. Jesus said, "If ye love me, keep my commandments" (John 14:15).
The good news of the gospel is that God does the work of obedience in the believer's life through His grace. "I can do all things through Christ which strengtheneth me" (Philippians 4:13). Through faith in God's promises, unbelievable power is released into the life of one who is willing to give up the life of sin. "We are more than conquerors through Him who loved us" (Romans 8:37).


text:
package com.crystalplanet.obsidianpoker.game;

public interface PlayerHandler {
    public void getNextAction(Player player);
}

redpajama_set_name: RedPajamaGithub


CBSI was founded by Bill J. Ulrich in April of 1998 specifically to meet the increasing clinical and financial challenges of tomorrow. Mr. Ulrich authored a PPS / Medicare manual for Skilled Nursing Facilities, writes a monthly Billing Advisor article for Provider magazine and has contracted to develop a model reimbursement system for the Washington Health Care Association. CBSI currently offers compliance programs to more than 25 long term-care facilities and files over 100 federal and state cost reports a year.


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Canada must actively recruit the best and brightest immigrants
In this Aug. 1, 2011 photo, a worker walks inside an office of Beijing Goldlink Go-Abroad Consulting Co., an immigration consulting firm assisting Chinese to immigrate to Canada and the United States.
Alexander F. Yuan/The Associated Press
Published May 4, 2012 Updated May 4, 2012
The world has changed, and when it comes to its immigration system, Canada is not changing fast enough to compete in it. It is no longer possible to sit back languidly, as the best and the brightest queue on its doorstep. The global market for human capital is voracious. There may always be migrants wanting to come to Canada, but they may not be the ones that Canada needs. People with options are less and less likely to tolerate hidebound and cumbersome immigration process, waiting as long as eight years to have their applications processed. If you are ambitious, if you are skilled, if you are entrepreneurial, if you are educated, if you are impatient for success, you will look elsewhere. Increasingly, elsewhere is looking better.
Countries like Australia can now fast-track applications for permanent residency in less than a year. Nor is the competition coming only from developed countries. With growing prosperity at home, not every upwardly mobile citizen of China, India and Brazil sees re-locating overseas as the only path to success. In fact, the Chinese and Indian governments are using investment, tax and visa incentives to draw the highly educated children of Chinese and Indian immigrants to their ancestral homelands.
To ensure Canada remains attractive to the sharpest minds, the keenest entrepreneurs and greatest innovators, the country must move beyond an inefficient selection system and long waits. Why should people put their careers on hold, in order to come to Canada? "International competition is starting to heat up for the best immigrants, the Frank Stronachs, the people who will drive the economy," notes Arthur Sweetman, an economist at McMaster University.
A new Gallup poll shows that Canada is the third most popular destination for people looking to relocate, with the U.S. first, and the U.K. second. Some might say that top three isn't bad. But Canada fell a spot from 2010, the global survey of 452,199 adults in 151 countries shows. Despite the recession, the U.S. remains by far the world's most desired destination for prospective migrants. Why isn't Canada in first place?
Immigration Minister Jason Kenney has enacted long-overdue reforms to streamline the selection system for economic immigrants. More points will be given for younger people with language proficiency who have prearranged employment.
He also plans to tackle the backlog by closing 100,000 files involving 300,000 people. This is a necessary measure – but one with consequences. It puts a stain on Canada's credibility, and more reforms are necessary to ensure the problem doesn't recur.
There is a need for a different kind of immigration officer to be sent to Canada's missions around the world: not someone with a shiny badge, armed with a long list of bureaucratic time-consuming checks that may end up impeding people with desirable educations, entrepreneurial instincts and in-demand skills from immigrating to Canada. Instead they should be people whose job it is to find and recruit talent. Canada needs headhunters.
Canada needs to open the doors for the right kind of migrant. Faster processing times would enable Canada to take advantage of global cyclical downturns. The current unemployment rate for Spaniards under age 25 is 50 per cent, the overall rate in the country is 25 per cent. Spain has an army of highly literate, technologically savvy people sitting idle, people who could, in some cases, literally walk in and fill vacant jobs here in the hi-tech, telecommunications, mining and petroleum sectors.
Instead of having a system flexible enough to seize on such opportunities, we have immigration lawyers with stories of clients – such as a couple from South Africa with MBAs – who become so frustrated, they simply give up on Canada.
Mr. Kenney is trying. Ottawa has expanded the provincial-nominee category, which favours immigrants with prearranged employment. However, the model has some weaknesses: Newcomers entering through the nominee stream are also less educated, and have lower salaries over the long term, than those who enter through the federal points system. This category of workers remains crucial to Canada's overall program, but it will not suffice.
Ottawa must do more to ensure newcomers can convert their foreign credentials and job experience. It must address discrimination in the labour market, and gate-keeping by professional associations. But first and foremost, Canada needs to change its mentality around immigration. It should be designed as much around whom Canada wants, as who wants Canada.
It is becoming a seller's market. As Sergio Karas, a Toronto immigration lawyer, says, "People from developing countries are no longer automatically migrating to Canada." Canada must learn to compete. Educated professionals, entrepreneurs, leaders, will not waste their most productive years trying just to get through the door. They know where they are wanted, and if they're not wanted here they will pack up their bags and go to where they are, taking with them all their potential and promise.
Why Canada needs a flood of immigrants
Editor's note: The immigration decisions we make today will define our future
Tell us: Nominate a change-maker in immigrant integration
Essential resources for immigrating to Canada
How a simple translation tool is helping doctors and immigrants communicate
How networking helped prevent an immigrant brain drain in Nova Scotia
Interactive: The case for the 400,000-immigrant solution
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I've been following the 'rise' of steampunk since 2009, when I realized that it combined two of my favorite things: historical fiction (umm, what my childhood was made of?) and science fiction. Best recent additions to the subgenre? Philip Reeve's Fever Crumb, Scott Westerfeld's Leviathan series, and Meljean Brooks' Iron Seas books. Of course for pure fun and fluff, Kady Cross' The Girl in the Steel Corset earns a nomination too. Imagine my delight when I saw this book cover and read the description. It sounds like I'll have another wonderful series to add to my reading list (joy!). Tiffany Trent's The Unnaturalists will be published by Simon & Schuster, and releases on August 14th, 2012.
That sounds really good! I love a good museum!
Up til now, I did not even know what steam-punk was. It totally does not sound anything like what it is.
I will have to pick up some and give them a shot, considering that I love both historical fiction and scifi.
I loved Leviathan (still need to read the last book though). I'll need to check out some more of these books you've recommended. I hope you get to read The Unnaturalists sometime soon.
Steampunk!!! I think the genre is new and is gaining good attention. I think if ever dystopian is old this genre will the be the next it! or is it the it already? lol but alas Unnaturalists sounds really good! Awesome pick!
Oooo, I was waiting for this one a few weeks ago. Great pick!
I'm finding myself more drawn to the historical genre and more often than not, steampunk is involved. It's just such a fascinating genre/time period to read about. I'm looking forward to reading about these unnatural creatures!
I should read steampunk, I don't know why I haven't been curious about it.
Fantastic choice! I'm a fan of steampunk in general, but this one in particular sounds like it could be really incredible. I adore the premise.
I've had this on my TBR list for a while now... so long, I didn't even realize they had a cover out for it! Beautiful!
That sounds so amazing! And steampunk! It's one of the best genres, and lots of fun! And the dresses, they look so pretty! Great pick! Thanks for stopping by! Happy reading!
SO excited about this one! I can't wait to read it.
Steampunk novels always have such pretty covers.
I think this one looks phenomenal! It has definitely made my waiting list too! Thanks for sharing your pick.
This sounds like an interesting pick. I've never heard of it before. Thanks for stopping by my blog!
This one sounds pretty awesome! I hope it's as good as the idea!


THE BRITISH OCCUPATION OF IRAQ: In the closing months of World War I, British forces defeated the Ottoman 6th Army, thus completing the occupation of Iraq. Approximately two years later, opposition to the British presence broke out into violence.
If only the American occupation had generated such ecumenical opposition!
Based on Tripp's account, it would seem that the British took the exact opposite approach to our own, which has been (until recently) to side with the Kurds and the Shi'ite majority against the Sunni elite.
What you think david? Is UK Rights in thier chose?
I'm not endorsing the idea, but one of the key logics of imperial divide-and-rule is to side with those who are most dependent upon you, usually the minority.
But David, judging by what we're doing with the Anbar Awakening folks under Sheikh al-Sitar, we're following the British path once again. The sheikhs might have formed the alliance in reaction to Al Qaeda but it speaks volumes when something goes wrong and they come running to the Americans and not the IG, whom they're officially working with.
The fact that we're arming them directly instead of through IG channels makes it fairly clear that we don't have any faith in the Shia government. It's just another form of divide-and-conquerism all over again.
Let me take a brief stab at the question posed by Anon 4:14. In terms of results, the British were obviously a step ahead of we are now. The revolt failed and Iraq required only limited attention from the British military thereafter.
Whether the British approach could work now is a different matter. I would say that a critical element of the British approach was the establishment of an Iraqi monarchy under a Hashemite ruler. Strange as it seems to us today, the idea of a new monarchy had considerable legitimacy in 1918.
In contrast, our only conception of legitimacy is democratic. Thus, it would've been very hard to side with anyone other than the Shi'ite majority, although we can make tactical compromises with the Sunnis, as Will points out.
In contrast, our only conception of legitimacy is democratic.
David, with due respect to your view, Iraq now far from democratic state! the fact is the government is prisoner in her Green Zone, Iraqis prisoner in their homes and the chaos any where their.
Talking about democracy in Iraq is just hot air.
May be what's came from Mahmoud Othman, a veteran Iraqi politician, put it, "The Americans are defeated. They haven't achieved any of their aims." so US lost what directions can take to settle things back in any way or state.
The Shiite man, who took Nivat around for her two weeks in Baghdad, in one of the more devastating quotes to come out of the capital in recent times, told her: "My uncles and cousins were murdered by Saddam's regime. I wanted desperately to get rid of him. But today, if Saddam's feet appeared in front of me, I would fall to my knees and kiss them!"
Anonymous 3:40 - I think you've misunderstood David's point. He was talking about the options open to an occupying force, based not simply on what was achievable but on what was thinkable given the political assumptions of the occupiers. The British in 1918 saw monarchy as a legitimate form of government, and could side with a minority in order to create it. The US in 2003 were committed to democracy, and that means you're pretty much obliged to work with the majority. In that respect, the US in 2003 had fewer options available to it than the British in 1918. Whether democratisation has been successful is a wholly different question, and I share some of your criticisms. But David's observation seems to me to be entirely correct.
Thank you, Gladstone. My point was certainly not that democratization has been successful, only that the United States' theoretical conception of legitimacy is limited to democratic options.
A different concept of legitimacy that is valid for many others is Islamic rule. Such conceptions of legitimacy range from hybrids of democracy and Islam to full-bore theocracy. To a limited extent, the new regime in Iraq incorporates certain Islamic concepts of validity.
What is entirely absent at that moment is a concept of legitimacy that is both secular and non-democratic. At times, nationalism, communism, socialism and military rule have all provided secular rationales for authoritarianism. The question remains, if democratization was futile in Iraq, what other options were available to the occupation government?
First thanks to Gladstone and David for your informative posts and reply.
But David, please I find it really fascinating when US officials or others from US talking about this mix of theirs as you put it in your replay above.
I use to be working with university and I met few American academics before 2003 and we talked about the war, I felt and I really believe more and more that most who rights about Iraq they judging Iraq as if they had in their mind Saudi and Gulf images, this is really sadden me.
Iraq society it's much different from the other Arab states in ME Iraq society was far open and developed in many aspects of life especially as a society contains different religious and ethnics groups which gave Iraq that richness fabric.
As you try to bring Islam and democracy here I think we need to understand we in ME and in Islamic world we have Religion "Islam" but heavily mixed with local culture which in my view that make the sickness with democracy not our Islamic law.
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As familiar as we may all think England to be to us outsiders, thanks to the prevalence of her language and culture around the globe, she does still tend to be somewhat foreign, when it comes right down to it. And the introductions of foreigners into an ecosystem can have dramatic consequences.
English week did precisely that to the 2. Bundesliga, delivering several surprise results in the middle of the week, with the trend continuing into the weekend's match day eight.
It doesn't always work, right Pauli fans?
Yet, there is a long-standing tradition of teams getting a bit of a lift after a coach is fired in the midst of a campaign.
1860 München divorced themselves from summer hire Ricardo Moniz after Tuesday's 1:0 loss at SV Sandhausen on an goal deep in extra time.
Three days after the disappointment in Sandhausen, the Lions, under the guidance of interim coach Markus van Ahlen grabbed just their second victory of the season, largely dominating SpVgg Gruether Fürth in a 2:0 Friday night fight.
Amid rumors of former Germany national player Bernd Schuster being the most-prominent candidate for the permanent gig, von Ahlen will continue his audition piece Friday night in Aalen.
If titles were awarded to sides for a three match burst, we'd be handing one to the Men in Violet right now.
Under new coach Tommy Stipic, Aue have seven points from their last three matches. Following up on the prior weekend's thrashing of St. Pauli, the Vielchen went to promotion candidate FC Ingolstadt and snagged a point in a 1:1 mid-week clash and then returned home Friday to take all three in a 1:0 with VfR Aalen.
Stipic thanking his Ingolstadt supporters.
Stipic, who joined Aue after a four-year stint in Ingolstadt training the U17 and reserve squads, was the recipient of chants and applause from the home supporters despite causing grief to Inglolstadt's hold on the top spot in the table.
Friday's victory was Aue's first over Aalen in seven attempts and leapfrogged the hosts over VfR in the table and out of the relegation zone, a feat that seemed, at best, a long way off, considering their rather moribund start to the season with just one goal and one point from their first five matches.
Vfr Aalen is now winless in their last six and will host the newly alive Lions of 1860 München Friday night.
Erzgebirge Barce . . . er . . . Aue head to Fürth for a showdown with the Kleeblätter of the SpVgg, also on Friday night.
VfL Bochum head coach Peter Neuerurer had been withholding a trip to the barber as long as his side was on their undefeated start to the season.
You want me to cut this?
1. FC Heidenheim apparently was not too pleased with how Neururer presented himself as a guest, giving Bochum's standing at the top of the 2. Bundesliga table a drastic hack job delivering a 5:0 buzzcut.
Before 20 minutes had elapsed Sunday, Mark Schnatterer had delivered a corner for Patrick Mayer to head home for the early lead, which Schnatter doubled from the spot when Mayer was taken down by Bochum's Timo Perthel. From there, the route was on, as Heidenheim put two more into the bank before the half and then mercifully added just one more before sending Bochum home with more to think about than just haircuts.
Heidenheim, now fourth in the table, will next travel to face fellow promoted side RB Leipzig in a top of the table showdown, while Bochum wil host crisis team 1. FC Nürnberg.
3 x 6 = promotion spot?
Everyone knows that a strong defense can anchor a successful season for a football club, so it should be no surprise that the three sides currently atop the second division table have in common the league's lowest goals-allowed total, each having surrendered just six goals over the first eight match days.
FC Ingolstadt surrendered one goal to Erzgebirge Aue Tuesday on what seemed like the only shot Aue managed to get anywhere near the target, forcing the table leaders to settle for a draw. Ralf Hasenhüttl's squad reconvened in Karlsruhe to snag a 0:1 road victory and, with it, a return to the top of the table, when Karlsruhe's Daniel Gordon was unable to direct his headed clearance of a Pascal Groß free kick somewhere other than his own goal.
RB Leipzig siezed second place over the two match days despite seeing the total goals allowed double after surrendering three goals total in their Wednesday 3:1 over Karlsruhe and a 2:2 at Düsseldorf. Yussuf Poulsen powered the victory over KSC with a pair of goals, but was forced to sit out the match at Fortuna due to a leg injury, opening the door for Georg Teigl to deliver a brace of his own.
The club practices of RB Leipzig were again the target of fan protests in their away match in Düsseldorf, with home fans borrowing the idea of wearing black to symbolize the death of fan culture in the club's home town.
It was another promoted side settling at table third, with SV Darmstadt collecting four points from a 4:0 victimization of a hapless FSV Frankfurt and a 1:1 at Union Berlin.
All four Darmstadt goals Wednesday came off corner kick play. Aytac Sulu delivered the finishing touch on two of them.
With a shot at ascending to the table top, Saturday's draw in the capitol city was a disappointment for the Lilien, as they largely dominated their hosts, limiting Union to just one shot on target before conceding the equalizer five minutes from time on a shot from 14 meters out.
The final match of the football-packed week arrived Monday with preseason promotion favorite 1. FC Nürnberg settled into a relegation spot as they were set to host 1. FC Kaiserslautern, who could spend the week in first place with a victory over their fellow Traditionsverein.
Yet, despite der Club having failed to score in any of their three prior matches and surrendering a total of eight to their three opponents in those matches, it was Valerien Ismael's side bolting to a 3:0 lead, with Alessandro Schöpf contributing a brace.
The Red Devils fought back with two goals by the 71st minute, but were unable to complete the comeback bid.
With Nürnberg struggles spurring rumors of a likely coaching change, the spectre of newly available taskmaster Felix Magath was made whole as the recent Fulham FC failure watched the match while seated next to former FCN president Gerd Schmelzer.
You can draw your own conclusions as to how motivation may have been spiked in this situation.


Home » Topics » Patient Monitoring » Research » Immune profiling of a patient with alemtuzumab-associated progressive multifocal leukoencephalopathy.
Summary of "Immune profiling of a patient with alemtuzumab-associated progressive multifocal leukoencephalopathy."
The spectrum of progressive multifocal leukoencephalopathy: A practical approach.
Bacterial and CMV pneumonia in a patient treated with alemtuzumab for multiple sclerosis.


Wouldn't it be great if you could browse, search, and insert photos from your Google account right to your blog posts and pages? Now you can.
Thank you! 🙂 I can kiss you right now. Alright, air kiss you.
This comes at the right time. I was just going to limit my recent photos to my Instagram and Pinterest because of ease. I actually think Google's is more user-friendly for what I generally want, so I'll look into this.
Thank you, we actually need this.
This is so awesome! So glad this is a feature now!
Thanks much. Now to really make me happy; Microsoft One Drive…PLEASE!
I was hoping this would soon be an option! Thank you for sharing this information!
Great news, thanks for supporting this feature into WordPress.
This doesn't work for me. If I would like to insert a picture from Google Photos for an article, I need to copy that, and it will take place in the storage. So, isn't it easier to simply upload a picture for my blog's WordPress media library? It would be better that if I upload something from Google Photos, I can simply insert them in the article without copying that, and nothing will take place in the storage.
There are many ways of achieving the same result. If your photos are already in Google Photos then this new feature will be the easiest and quickest way to use them on your blog, and you can do this while still editing your post. If you have experienced a problem then please contact us.
This is a great idea – once you get it up and working. I just went to my Media Library – as per your instructions – and there is no option to connect my Google account! Why is this?
Google is an option from the dropdown menu in the top left corner of the Media Library. Full details are on our support page, and you can contact support for further help.
It seems like a good idea, however I am not too sure how I would use it as presumably I would be obliged to select from photos chosen by Google. I already have a stack of photos in my media library and googling does not take long. Does the new system involve waiting for many photos to download or do you go to google to choose?
This provides access to your own photos, stored on Google Photos. It does not provide access to Google's image search.
Will this feature be included in Jetpack any time in the future for those of us with self-hosted sites?
Yep it's already available for Jetpack sites! Just pick your Jetpack-powered site from WordPress.com and you'll have the same Google option in the Media Library.
Went to the Dashboard, went to the library, couldn't find the Google Photo link. Odd.
Oh. This is good!! I usually google my own blog's images and copy but this is better. Thank you!
Will photos from Google still count towards storage space?
I'm using self-hosted WordPress version 4.8.2 and don't see this option in the upper left of the Media Library. Is there a particular plugin that needs to be enabled?
Sounds excellent, even though I don't use Google Photos, and never knew it existed, but now is the time to discover all about it and start using it!
This is going to make posting images so much quicker and easier, thank you WordPress and Google Photos!
Awesome! Thumbs up to the WordPress.com team…..
I've been wanting this forever! Thanks WordPress!
Pretty neat feature!. Just curious, will this feature also be available to those that use flickr in the future?.
We're looking at supporting other services.
Excellent feature, but will it be available directly from self hosted sites without using WordPress.com? This cuts out so many steps in our publishing workflow, but then adds several more if we then need to login to WordPress to do this. Our admin setup is complex, so switching to editing here isn't an option.
I've multiple contributors who write directly and this would be a game changer if they could directly upload their images from their account.
It is only available through WordPress.com / Jetpack, and there are no plans for a standalone version.
This makes blogging easy for everyone. I love to use WordPress.
This sounds useful. I have a lot of photos that I put on Google over the years. And I could go in and download the ones I am interested in, but this seems like an easy way to connect. That said, and call me cautious, but when a user connects to their Google account, what is Google then enabled to see of what's on the WordPress account and what is WordPress enabled to see of what's in the user's Google account?
The connection is one-way – Google has no path back to WordPress. When you connect to Google it lists the exact permissions being requested and you need to confirm these, which in this instance is specifically just access to your Google Photos. You can see this on the support page. You can disconnect at any point from WordPress.com or Google and any access is instantly revoked.
I'm doing a happy dance right now! Thank you!
Yes, as mentioned above you will need Jetpack. Once everything is installed and working you will be able to edit your self-hosted WordPress from WordPress.com and have access to your Google photos.
Awesome new feature! One wrinkle: My photos on Google are all at native high-res. I typically re-scale my photos before posting to speed up load times and take less bandwidth, as a lot of my readers on on mobile. Is there any future plan to include a simple photo editor to resize or crop photos before posting from Google photos?
You can already use the WordPress.com editor to crop and resize your images. Newer themes will also typically load a smaller version of the image for mobile users.


Ryan Thomas shares an update on reuniting with his father and how there is HOPE AFTER ALIENATION!
Shared Parenting is possible! An For some, this may be the experience with your co-parent and children. For others, this may be a long awaited dream. My hope is that all children have this experience. To know that both parents want and love them and that both parents work together to make this happen for their children out of pure parental LOVE!
Anonymous: Hey guys, I am a child of divorce. You can share my story if you want, just leave my name off. My parents divorced when I was 4. my dad was an alcoholic and was violent towards my mother. He did get clean tho and stopped that behavior. My mom had hundreds of reasons to be bitter towards my dad..hundreds of them! She could have hated him for all he did to her and for all he put her through (and maybe she did but she never let that on to my sister and I) and could have kept him from us. My mom got full custody of me and my sister, with my dad paying child support, however I was allowed to be with him whenever I wanted. My mom never told me no. I had a bedroom at his house and a bedroom at my moms. If I felt like staying at moms, I did, and vice versa. Not once did I think my dad's house wasn't my home. They were both my home! I was never living out of a suitcase. I had two parents who both loved me and two parents that I equally needed in my life. I am so happy that they could put aside their differences and make it work for me and my sister!
Exchanges were no big deal. As an adult, I know now they didn't exactly like seeing each other but I would have never guessed that as a child! They were all smiles. They acted exactly the way adults should act!! When the opposition says that a child needs one home, I shake my head. Kids are so resilient and adapt so easily. Having one home with mom and one home with dad isn't a problem to them…they adjust. Might there be a trial period, of course, but they will adapt!!
As far as 2 adults not being able to get along so shared parenting won't work..I say this! Grow up! You chose to have a child with somebody so be adult enough to raise your child with them!! I would resent my mom today if she wouldn't have allowed me as much time with my father as I wanted! A child having a bond with each parent is so so so important and one parent shouldn't get in the way of that just because they don't like their ex!! Shared parenting is definitely the way to go!!
Thank you for sharing this uplifting and heart felt story!
Note: Spacing to create paragraphs was added. The words and order of the story are presented as submitted.


As their names suggest, no credit check loans are loans that don't involve credit checks unlike normal loans you'll obtain from the banks or any other high street lenders. The purpose of these loans is to offer an alternative to traditional lending, where not everyone may qualify, especially those who have less than perfect credit.
You may need a no credit check loan for the most obvious reason, and that is because you may have bad credit and cannot get accepted for personal loans. It pays to know, however, that not everyone who got refused a loan from banks and other lending institutions necessarily have indecent credit. It could be that you are a newly college grad or a new employee, and have no previous experience with credit yet. It could also be that you are self-employed and are not receiving wage slips every 15 to 30 days. If this is you, you can benefit from a loan with no credit checks.
Additionally, a no credit check loan allows you to obtain cash fast. The benefit of skipping credit checks is that not only can you qualify despite having bad credit, but it makes everything a lot faster compared to getting a loan from the high street. This means very little paperwork to accomplish, and no waiting for days or weeks to get your money.
No credit check loans may either be short or long term, which can be payable between two weeks and 36 months. They can also either be unsecured or secured by collateral. A perfect example of a secured loan is a logbook loan, where the borrower's vehicle serves as collateral against the loan.
Lets you borrow more. Because a logbook loan is essentially backed up by security, you are given access to more cash, which you can release through your vehicle. Unlike other no credit check loans which limit you to about £500 to £1,000 only, with a logbook loan you can borrow as much as £10,000! The more value your vehicle has, the more money you can unleash!
More flexible repayment terms. Unlike other loans for bad credit such as payday loans, you can pay the loan in instalments, instead of a one lump sum. Additionally, the length of the loan is more flexible, which can last anywhere between three months to three years, depending on the amount you borrowed.
Available for self-employed. While other loans may look closely into your employment history, a logbook loan only requires that you have a stable and sustainable income source. You don't need to worry about having to present pay stubs if you are self-employed and don't have them, because your bank statement will be enough proof.
Surely it will always depend on your own individual circumstances, but if you own a working vehicle that has some value in it, you might want to consider a logbook loan to help you raise that quick cash that you need.


charged as well as the length of your loan. The higher your score, the more likely you'll be approved for a consumer loan and receive favorable terms.
In these pressing economic times, not everyone has a good FICO credit score, which can be especially problematic if you need to apply for a consumer loan. Whether seeking a mortgage, a home equity loan/line of credit, car loan, credit card, or some other type of loan, you need to get the highest score possible.
Shrink those balances: You don't have to pay off your credit cards, but running big balances is a red flag to creditors. Work on reducing your debt, a step which will gradually raise your credit score.
Don't apply for too many loans: You may have unwittingly caused your credit score to drop by applying for too many loans in a short period of time. This can happen if you are planning to shop for a new car and are arranging your own financing. By applying to several different lending institutions for the sake of finding the best deal, you'll be shooting up another warning flag to creditors. Find out the rate first, then apply.
Remedy credit problems: If you've been late making payments in the past, then your score will take a hit. Make payments on time and pay more than the minimum amount due each month. Get free copies of your credit reports and check them for errors; notify the credit reporting agencies if you find mistakes. They are required by law to fix mistakes within thirty days or that information must be automatically removed from your credit report.
Keep consumer accounts open: Odd as it may sound, closing a credit card or other consumer account will negatively impact your credit score. Simply tuck your unused credit cards away in a safe place and don't use them again. You can gradually close them after you secure new credit, especially if you have no plans to borrow again in the near future.
More accounts means a reduced score: Opening more accounts will work against you. Only open up enough consumer accounts as needed.
Consider NOT moving your money around: Consumers have gotten into the habit of shifting outstanding balances from one account to another, but that move can actually reduce your credit score. Consolidating your balances to one account may cause your credit score to drop.
Building a good credit history is an achievable and laudable goal for any consumer. Take care of your credit score and your credit score will take care of you in the form of favorable lending terms for your next consumer lending opportunity.


Put Chana dal, ginger and green chillies in a mixer and make a coarse paste.
Add mashed boiled banana, 1/2 tsp chilli powder, 1/2 tsp roasted cumin powder, salt. Make very smooth batter.
Put 1-2 drops of oil in each cavity of appam pan and heat it.
Pour this batter in all the cavities. Let it cook on medium flame.
When it is cooked from one side, flip it and let it cook from other side as well. Add 1-2 drops of oil if required.
Mix curd, 1 tsp sugar and salt very well. Curd shouldn't be very thick.
Now soak all the vadas in this curd for about 2 hours.
You will notice that vadas are bigger in size now.
Add Tamarind chutney, red chilli powder and roasted cumin powder. Garnish it with coriander leaves.


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