Robotic extraction of DNA from dilutions of blood and semen using either the BioRobots® EZ1 or BioRobots® M48 consistently produced lower recoveries than standard organic extractions of the same ...samples. In an effort to increase the efficiency of robotically extracted DNA, glycogen and carrier RNA were added following cell lysis. The addition of glycogen, postlysis, resulted in no improvement in DNA recovery with the BioRobot® EZ1. However, when carrier RNA was added to the cell lysate of limited and degraded samples extracted on the EZ1 or the M48, DNA recoveries dramatically increased four‐ to 20‐fold. DNA yields obtained by robotic extraction in the presence of carrier RNA were as high, or higher, as those obtained by organic extraction lacking carrier RNA, while experiments that utilized carrier RNA in both types of extractions showed increased sensitivity for both methods. Furthermore, carrier RNA substantially increased the recovery of fragmented DNA with the EZ1.
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BFBNIB, FZAB, GIS, IJS, KILJ, NLZOH, NUK, OILJ, SBCE, SBMB, UL, UM, UPUK
2.
Essential Role for RGS9 in Opiate Action Zachariou, Venetia; Georgescu, Dan; Sanchez, Nick ...
Proceedings of the National Academy of Sciences - PNAS,
11/2003, Volume:
100, Issue:
23
Journal Article
Peer reviewed
Open access
Regulators of G protein signaling (RGS) are a family of proteins known to accelerate termination of effector stimulation after G protein receptor activation. RGS9-2, a brain-specific splice variant ...of the RGS9 gene, is highly enriched in striatum and also expressed at much lower levels in periaqueductal gray and spinal cord, structures known to mediate various actions of morphine and other opiates. Morphine exerts its acute rewarding and analgesic effects by activation of inhibitory guanine nucleotide-binding regulatory protein-coupled opioid receptors, whereas chronic morphine causes addiction, tolerance to its acute analgesic effects, and profound physical dependence by sustained activation of these receptors. We show here that acute morphine administration increases expression of RGS9-2 in NAc and the other CNS regions, whereas chronic exposure decreases RGS9-2 levels. Mice lacking RGS9 show enhanced behavioral responses to acute and chronic morphine, including a dramatic increase in morphine reward, increased morphine analgesia with delayed tolerance, and exacerbated morphine physical dependence and withdrawal. These findings establish RGS9 as a potent negative modulator of opiate action in vivo, and suggest that opiate-induced changes in RGS9 levels contribute to the behavioral and neural plasticity associated with chronic opiate administration.
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BFBNIB, NMLJ, NUK, PNG, SAZU, UL, UM, UPUK
Short tandem repeat (STR) analysis is a valuable tool in identifying the source of biological stains, particularly from the investigation of sexual assault crimes. Difficulties in analysis arise ...primarily in the interpretation of mixed genotypes when cell separation of the sexual assailant's sperm from the victim's cells is incomplete. The forensic community continues to seek improvements in cell separation methods from mixtures for DNA typing. The feasibility of applying laser microdissection (LMD) technology to precisely separate sexual assault cell mixtures by visual inspection coupled with laser dissection was assessed through three experiments. First, various histological stains were evaluated for use with LMD and DNA analysis. Second, different DNA isolation methods were evaluated on LMD‐collected cells. Finally, STR analysis was performed on LMD‐separated sperm cells from mixtures of semen and female buccal epithelial cells. The results indicated (a) hematoxylin/eosin staining performed best in its ability to differentiate sperm and epithelial cells while exhibiting the least negative effect on further downstream analysis; (b) both QIAamp® and Lyse‐N‐Go™ methods were useful for recovery of DNA from LMD‐collected sperm cells; and (c) LMD separation provided clear STR profiles of the male donor with the absence of any additional alleles from the female donor. This report describes an efficient, low‐manipulation LMD method for the efficient separation of spermatozoa from two‐donor sperm/epithelial cell mixtures.
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BFBNIB, FZAB, GIS, IJS, KILJ, NLZOH, NUK, OILJ, SBCE, SBMB, UL, UM, UPUK
Schizophrenia is a heritable disorder with substantial public health
impact. We conducted a multi-stage genome-wide association study (GWAS) for
schizophrenia beginning with a Swedish national sample ...(5,001 cases, 6,243
controls) followed by meta-analysis with prior schizophrenia GWAS (8,832 cases,
12,067 controls) and finally by replication of SNPs in 168 genomic regions in
independent samples (7,413 cases, 19,762 controls, and 581 trios). In total, 22
regions met genome-wide significance (14 novel and one previously implicated in
bipolar disorder). The results strongly implicate calcium signaling in the
etiology of schizophrenia, and include genome-wide significant results for
CACNA1C
and
CACNB2
whose protein products
interact. We estimate that ∼8,300 independent and predominantly common
SNPs contribute to risk for schizophrenia and that these collectively account
for most of its heritability. Common genetic variation plays an important role
in the etiology of schizophrenia, and larger studies will allow more detailed
understanding of this devastating disorder.
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DOBA, IJS, IZUM, KILJ, NUK, PILJ, PNG, SAZU, UILJ, UKNU, UL, UM, UPUK
...95 per cent of organizations defined as "engaged" use annual employee surveys, compared with 45 per cent in disengaged organizations, according to a 2016 survey conducted by Quantum Workplace. ......focus on the data points that are most important to the company - be they around employee development or areas of company culture that are lagging.
Lineage conversion of differentiated cells in response to hormonal feedback has yet to be described. To investigate this, we studied the adrenal cortex, which is composed of functionally distinct ...concentric layers that develop postnatally, the outer zona glomerulosa (zG) and the inner zona fasciculata (zF). These layers have separate functions, are continuously renewed in response to physiological demands, and are regulated by discrete hormonal feedback loops. Their cellular origin, lineage relationship, and renewal mechanism, however, remain poorly understood. Cell-fate mapping and gene-deletion studies using zG-specific Cre expression demonstrate that differentiated zG cells undergo lineage conversion into zF cells. In addition, zG maintenance is dependent on the master transcriptional regulator Steroidogenic Factor 1 (SF-1), and zG-specific Sf-1 deletion prevents lineage conversion. These findings demonstrate that adrenocortical zonation and regeneration result from lineage conversion and may provide a paradigm for homeostatic cellular renewal in other tissues.
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•Homeostasis and regeneration involve lineage conversion of differentiated cells•Glomerulosa cell maintenance is dependent on the core transcriptional regulator SF1•An alternative pathway maintains zF cell production after loss of SF1 in zG cells
Freedman et al. show that adrenocortical zonation during postnatal development, adult homeostasis, and tissue regeneration involves lineage conversion between differentiated cells in response to hormonal feedback. These findings underscore the plasticity of this vital organ and suggest that other adult tissues may also depend on lineage conversion for their maintenance.
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GEOZS, IJS, IMTLJ, KILJ, KISLJ, NLZOH, NUK, OILJ, PNG, SAZU, SBCE, SBJE, UILJ, UL, UM, UPCLJ, UPUK, ZAGLJ, ZRSKP
Objective
To estimate the prevalence of amyloid positivity, defined by positron emission tomography (PET)/cerebrospinal fluid (CSF) biomarkers and/or neuropathological examination, in primary ...progressive aphasia (PPA) variants.
Methods
We conducted a meta‐analysis with individual participant data from 1,251 patients diagnosed with PPA (including logopenic lvPPA, n = 443, nonfluent nfvPPA, n = 333, semantic svPPA, n = 401, and mixed/unclassifiable n = 74 variants of PPA) from 36 centers, with a measure of amyloid‐β pathology (CSF n = 600, PET n = 366, and/or autopsy n = 378) available. The estimated prevalence of amyloid positivity according to PPA variant, age, and apolipoprotein E (ApoE) ε4 status was determined using generalized estimating equation models.
Results
Amyloid‐β positivity was more prevalent in lvPPA (86%) than in nfvPPA (20%) or svPPA (16%; p < 0.001). Prevalence of amyloid‐β positivity increased with age in nfvPPA (from 10% at age 50 years to 27% at age 80 years, p < 0.01) and svPPA (from 6% at age 50 years to 32% at age 80 years, p < 0.001), but not in lvPPA (p = 0.94). Across PPA variants, ApoE ε4 carriers were more often amyloid‐β positive (58.0%) than noncarriers (35.0%, p < 0.001). Autopsy data revealed Alzheimer disease pathology as the most common pathologic diagnosis in lvPPA (76%), frontotemporal lobar degeneration–TDP‐43 in svPPA (80%), and frontotemporal lobar degeneration–TDP‐43/tau in nfvPPA (64%).
Interpretation
This study shows that the current PPA classification system helps to predict underlying pathology across different cohorts and clinical settings, and suggests that age and ApoE genotype should be considered when interpreting amyloid‐β biomarkers in PPA patients. Ann Neurol 2018;84:737–748
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BFBNIB, FZAB, GIS, IJS, KILJ, NLZOH, NUK, OILJ, SBCE, SBMB, UL, UM, UPUK
A power transmission system can be represented by a network with nodes and links representing buses and electrical transmission lines, respectively. Each line can be given a weight, representing some ...electrical property of the line, such as line admittance or average power flow at a given time. We use a hierarchical spectral clustering methodology to reveal the internal connectivity structure of such a network. Spectral clustering uses the eigenvalues and eigenvectors of a matrix associated to the network, it is computationally very efficient, and it works for any choice of weights. When using line admittances, it reveals the static internal connectivity structure of the underlying network, while using power flows highlights islands with minimal power flow disruption, and thus it naturally relates to controlled islanding. Our methodology goes beyond the standard k-means algorithm by instead representing the complete network substructure as a dendrogram. We provide a thorough theoretical justification of the use of spectral clustering in power systems, and we include the results of our methodology for several test systems of small, medium and large size, including a model of the Great Britain transmission network.
One of the most widely conserved hallmarks of aging is a decline in functional capabilities. Mobility loss is particularly burdensome due to its association with negative health outcomes, loss of ...independence and disability, and the heavy impact on quality of life. Recently, a new condition, physical frailty and sarcopenia, has been proposed to define a critical stage in the disabling cascade. Physical frailty and sarcopenia are characterized by weakness, slowness, and reduced muscle mass, yet with preserved ability to move independently. One of the strategies that have shown some benefits in combatting mobility loss and its consequences for older adults is physical activity. Here, we describe the opportunities and challenges for the development of physical activity interventions in people with physical frailty and sarcopenia. The aim of this article is to review age-related physio(patho)logical changes that impact mobility in old age and to provide recommendations and procedures in accordance with the available literature.
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