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Reactivating Fetal Hemoglobin Expression in Human Adult Erythroblasts Through BCL11A Knockdown Using Targeted Endonucleases

We examined the efficiency, specificity, and mutational signatures of zinc finger nucleases (ZFNs), transcriptional activator-like effector nucleases (TALENs), and clustered regularly interspaced short palindromic repeat (CRISPR)/Cas9 systems designed to target the gene encoding the transcriptional repressor BCL11A, in human K562 cells and human CD34+progenitor cells. ZFNs and TALENs were delivere

Immunocytochemical localisation of neuronal nitric oxide synthase in developing and transplanted rabbit retinas

Nitric oxide (NO) acts as a modulator of neuronal transmission in mature neuronal systems, including the retina. Recently, NO has also been suggested to have a trophic function during development. We examined immunocytochemically the distribution of NO-producing cells in developing and transplanted rabbit retinas. An antibody detecting the neuronal isoform of its biosynthetic enzyme, nitric oxide

Stimulation-evoked release of purines from the rabbit retina

The evoked release of purines from rabbit retinae preloaded with [(3)H]adenosine was studied in vitro. Potassium (8.6-43.6 mM) and ouabain (1 or 10 microM) increased the release of radioactivity in a concentration-dependent manner. The K(+)-evoked release was significantly reduced when the superfusion was carried out at 2-4 degrees C. The effect of K(+) (8.6, 13.6 and 23.6 mM) and of ouabain (1 mi

Localization of enkephalinergic neurons in the central nervous system of the salmon (Salmo salar L.) by in situ hybridization and immunocytochemistry

The distribution of neurons expressing preproenkephalin (PPE) mRNA in the brain of the salmon was investigated by means of non-radioactive in situ hybridization, and directly compared with the distribution of enkephalin-immunoreactive (ENKir) neurons. This approach, utilized here for the first time in a non-mammalian vertebrate for the identification of neurons containing opioid peptides, permitte

In situ hybridization of neuropeptide Y (NPY) mRNA in the goldfish brain

The distribution of neurones expressing neuropeptide Y (NPY) mRNA in the brain of the goldfish was investigated using a non-radioactive in situ hybridization technique. Neurones expressing NPY mRNA were located in cell groups previously shown to exhibit NPY immunoreactivity in the ventral telencephalon, thalamus, optic tectum and a region adjacent to the locus coeruleus. In addition, neurones expr

Expression of brain-derived neurotrophic factor and of its functional receptor in neonatal and adult rat retina

The expression of mRNA coding for brain-derived neurotrophic factor (BDNF) and for its functional receptor, the full-length tyrosine kinase receptor trkB (trkB mRNA), was examined in early postnatal and adult rat retina by in situ hybridization using digoxygenin and radioactively-labeled oligonucleotide probes. BDNF and trkB mRNAs are expressed in the ganglion cell layer at postnatal-days (PN) 1,

Localisation of neuronal nitric oxide synthase-immunoreactivity in rat and rabbit retinas

The distribution of neuronal nitric oxide synthase (NOS) immunoreactivity was examined in rat and rabbit retinas and was compared with the distribution of nicotinamide adenine dinucleotide phosphate (NADPH)-diaphorase reactivity and vasoactive intestinal peptide (VIP) immunoreactivity. An antibody raised against a C-terminal fragment of a cloned rat cerebellar NOS was used to localise NOS immunore

Distribution of GABA immunoreactivity in kainic acid-treated rabbit retina

Ischaemic retinal cell degeneration seems to involve both NMDA and non-NMDA receptor over-stimulation. However, different retinal cell types differ largely in their susceptibility to excitatory amino acid-induced neurotoxicity. We have investigated the vulnerability of GABAergic cells in the rabbit retina to the non-NMDA receptor agonist kainic acid (KA). The distribution of GABA immunoreactivity

Colocalization of (3H)-adenosine accumulation and GABA immunoreactivity in the chicken and rabbit retinas

Using combined autoradiography and immunohistochemistry, we have compared (3H)-adenosine accumulation and GABA immunoreactivity in the chicken and rabbit retinas. Colocalization of the two markers was observed in a subset of amacrine cells and in certain cell bodies in the ganglion cell layer in both species and in a few horizontal cells in the chicken retina. Cells that contained only (3H)-adenos

Release of endogenous and radioactive purines from the rabbit retina

The adenine nucleotide pool of rabbit retina was labeled by an intravitreal injection in vivo of [3H]adenosine. Practically all the radioactivity was retained in the form of adenine nucleotides. The relative proportion of [3H]adenine nucleotides was the same as that of endogenous nucleotides. Potassium depolarization (43.6 mM) in vitro caused a rapid increase in the rate of release of radioactive

Autoradiography of nucleoside uptake into the retina

A selective uptake mechanism for some nucleosides and related substances was found in retinae of light adapted rabbits and fish. After the intravitreal injection in vivo of [(3)H]adenosine, [(3)H]inosine, [(3)H]guanosine and certain related compounds, the distribution of radioactivity was studied by autoradiography. Retinae were also incubated in [(3)H]adenosine and [(3)H]inosine and then were sim

Adenosine in vertebrate retina : localization, receptor characterization, and function

1. The uptake of [3H] adenosine into specific populations of cells in the inner retina has been demonstrated. In mammalian retina, the exogenous adenosine that is transported into cells is phosphorylated, thereby maintaining a gradient for transport of the purine into the cell. 2. Endogenous stores of adenosine have been demonstrated by localization of cells that are labeled for adenosine-like imm

The Construction of the "I" in the Lord's Prayer : A Social-constructive Discourse Analysis

This paper addresses how the “I” of the praying individual is constructed in the interaction with the divinity in the praying of the Lord’s Prayer. This approach draws on social-constructive theory on identity formation and developmental psychology and a close reading of the Lord’s Prayer in Matthew 6 and the Sermon on the Mount. It is the assertion of this paper that a discourse analysis of the L

Using green infrastructure for urban climate-proofing : An evaluation of heat mitigation measures at the micro-scale

Urban green infrastructure (UGI) has been increasingly promoted as a key measure to mitigate heat stress in cities caused by the urban heat island effect and climate change impacts, including climate variability and extremes. However, comparable information concerning the performance of different UGI types to moderate such impacts is mostly lacking. This creates serious challenges for urban planne

Provenance does matter : links between winter trophic segregation and the migratory origins of European robins

Amongst migratory species, it is common to find individuals from different populations or geographical origins sharing staging or wintering areas. Given their differing life histories, ecological theory would predict that the different groups of individuals should exhibit some level of niche segregation. This has rarely been investigated because of the difficulty in assigning migrating individuals

Assembly of tapasin-associated MHC class I in the absence of the transporter associated with antigen processing (TAP)

The assembly of MHC class I molecules is regulated by a multi-protein complex in the endoplasmic reticules (ER) termed the loading complex. Tapasin is suggested to be one of the molecules forming this complex on the basis of its interaction with both the transporter associated with antigen processing (TAP) and MHC class I molecules. To address whether TAP is indispensable for the processing of the

Cerebrospinal fluid-induced retardation of amyloid β aggregation correlates with Alzheimer's disease and the APOE ε4 allele

Misfolding and aggregation of amyloid β (Aβ) are key features of Alzheimer's disease (AD) pathogenesis, but the molecular events controlling this process are not known in detail. In vivo, Aβ aggregation and plaque formation occur in the interstitial fluid of the brain extracellular matrix. This fluid communicates freely with cerebrospinal fluid (CSF). Here, we examined the effect of human CSF on A

From surfactant to cellulose and DNA self-assembly. A 50-year journey

Surfactants have been the basis for applications in several industrial sectors for a long time. However, fundamental research was 50 years ago still limited to a small number of academic groups and even basic aspects were controversial. The field has since undergone an enormous expansion and the improved understanding has laid the basis of numerous new products as well as been the basis of importa