Synthetic hydrogels formed by thiol-ene crosslinking of vinyl sulfone-functional poly(methyl vinyl ether-alt-maleic acid) with α,ω-dithio-polyethyleneglycol.

Polymer hydrogels formed by rapid thiol-ene coupling of macromolecular gel formers can offer access to versatile new matrices. This paper describes the efficient synthesis of cysteamine vinyl sulfone (CVS) trifluoroacetate, and its incorporation into poly(methyl vinyl ether-alt-maleic anhydride) (PMMAn) to form a series of CVS-functionalized poly(methyl vinyl ether-alt-maleic acid) polymers (PMM-CVSx) containing 10 to 30 mol% pendant vinyl sulfone groups. Aqueous mixtures of these PMM-CVS and a dithiol crosslinker, α,ω-dithio-polyethyleneglycol (HS-PEG-SH, Mn = 1 kDa), gelled through crosslinking by Michael addition within seconds to minutes, depending on pH, degree of functionalization, and polymer loading. Gelation efficiency, Young's modulus, equilibrium swelling and hydrolytic stability are described, and step-wise hydrogel post-functionalization with a small molecule thiol, cysteamine, was demonstrated. Cytocompatibility of these crosslinked hydrogels towards entrapped 3T3 fibroblasts was confirmed using a live/dead fluorescence assay.

Effects of electromagnetic fields emitted by GSM phones on working memory: a meta-analysis.

BACKGROUND AND OBJECTIVE: Current treatments for Alzheimer's Disease (AD) do not affect the course of the illness and brain stimulation techniques are increasingly promoted as potential therapeutic interventions for AD. This study reviews the effects of electromagnetic field (EMF) exposure versus sham exposure on working memory (WM) performance of healthy human participants. METHOD: Online literature databases and previous systematic reviews were searched for studies of EMF and WM in participants without reported memory problems. Two thousand eight hundred and fifty seven studies were identified, and 10 studies met the inclusion criteria. An assessment of study quality was completed, and separate, random effects meta-analyses were conducted for each of the three WM tasks included: n-back, substitution and digit span forward. RESULTS: No differences were found between participants exposed to active EMF versus sham conditions in any of the three working memory tasks examined. CONCLUSION: Results indicate that EMF does not affect WM during the n-back, substitution and digit-span tasks. Future studies should focus on the possible effects of chronic exposure to EMF in older adults with AD using a battery of comparable WM and attention tasks, before EMF can be seriously considered as a potential modulator of WM in AD. Copyright © 2016 John Wiley & Sons, Ltd.

DNA barcoding of Canada's skates.

DNA-based identifications have been employed across broad taxonomic ranges and provide an especially useful tool in cases where external identification may be problematic. This study explored the utility of DNA barcoding in resolving skate species found in Atlantic Canadian waters. Most species were clearly resolved, expanding the utility for such identification on a taxonomically problematic group. Notably, one genus (Amblyraja) contained three of four species whose distributions do not overlap that could not be readily identified with this method. On the other hand, two common and partially sympatric species (Little and Winter skates) were readily identifiable. There were several instances of inconsistency between the voucher identification and the DNA sequence data. In some cases, these were at the intrageneric level among species acknowledged to be prone to misidentification. However, several instances of intergeneric discrepancies were also identified, suggesting either evidence of past introgressive hybridization or misidentification of vouchered specimens across broader taxonomic ranges. Such occurrences highlight the importance of retaining vouchered specimens for subsequent re-examination in the light of conflicting DNA evidence.

Spontaneous association of glial cells with regrowing neurites in mixed cultures of dissociated spiral ganglia.

Evidence from developmental and regeneration studies of the cochlea and other tissues gives reason to hypothesize a role for nonneural cells in the growth and regeneration of cochlear spiral ganglion nerve fibers. We examined the spontaneous associations of regrowing neurites and nonneural cells in mixed cultures of dissociated newborn mouse spiral ganglia. After 7 days in vitro, nonneural cells formed a confluent layer in the culture well. Regrowing neurites grew atop this layer, forming non-uniform patterns that were similar to those formed by endogenously expressed laminin-1, entactin and integrin beta4, but not fibronectin or tenascin. In cultures grown for 42 h and maintained in three different growth media, all regrowing neurites were preferentially associated with spindle-shaped nonneural cells. The spindle-shaped cells incorporated bromodeoxyuridine in culture and were immunoreactive for the proteins S100, laminin-1, laminin-2, SRY-related high-mobility-group box 10 transcription factor (Sox10), neurotrophin receptor (P75) and connexin29 but negative for fibronectin and glial fibrillary acidic protein. These cells existed in the culture within a much larger, general population of fibronectin positive cells. Immunolabeling of fixed cochleas from neonatal mice localized Sox10, P75 and connexin29, to peripheral nerve bundles. The observed expressions of protein markers and the bipolar, spindle shape of the neurite-associated cells indicate that they are derived in vitro from the original Schwann or satellite cells in the ganglion or spiral lamina. The spontaneous and preferential association of neurites in culture with mitotic Schwann cells highlights the potential contribution neurite-Schwann cell interactions may have in promoting the growth and regrowth of damaged spiral ganglion neurons in the cochlea.

Culture conditions determine the prevalence of bipolar and monopolar neurons in cultures of dissociated spiral ganglion.

To gain insight into the mechanisms that control the generation or maintenance of the characteristic bipolar morphology of cochlear spiral ganglion neurons, we have taken advantage of our recently developed procedure for culture of dissociated newborn mouse spiral ganglion. In these cultures, inclusion of the cytokine leukemia inhibitory factor (LIF) in the medium increases neuronal survival and the number of bipolar neurons. Here we tested effects of two other LIF-type cytokines (ciliary neurotrophic factor, CNTF; and human recombinant oncostatin M, hOSM) and of bone morphogenetic protein 4 (BMP4) on survival, morphology and neurite lengths of neurons in cultures of dissociated spiral ganglion. Like LIF, CNTF and hOSM increased neuronal survival and the number of surviving bipolar neurons. BMP4 also increased neuronal survival, but unlike LIF, CNTF and hOSM, increased the number of monopolar neurons and neurons with no neurites. In addition, population histograms demonstrate that the population lengths of the longer and shorter neurites of bipolar neurons were shorter in BMP4 containing cultures than in control or LIF cultures. When LIF and BMP4 were simultaneously added to the cultures, the BMP4 effects predominated. These experiments demonstrate that exposure to different environmental conditions can result in different morphologies in the surviving population of spiral ganglion neurons in culture.

Survival and morphology of auditory neurons in dissociated cultures of newborn mouse spiral ganglion.

We have systematically characterized neuronal survival and growth in cultures derived from newborn/postnatal day 1 mouse cochlea. Dissociated cultures of the cochlear spiral ganglion provide an experimental environment in which to examine molecular mechanisms of survival, development and physiology of auditory neurons. To relate survival to the total number of neurons present in the source tissue, three cochleas from different newborn CD-1 mice were embedded in Araldite resin and serially sectioned at 5 mum thickness. All neurons were counted. To avoid overcounting, each section served as a lookup section for the next, giving 8240+/-423 (S.D.) neurons per ganglion. Cultures maintained in the presence of adjacent non-neural tissue, brain-derived neurotrophic factor, neurotrophin 3, leukemia inhibitory factor (LIF) and 10% fetal bovine serum returned the best overall survival (30%) at 42 h post-plating. Best overall survival required the continuous presence of a serum component(s) larger than 100,000 MW. Plating efficiency (number of neurons that attach to the well after 4 h) was similar in the presence or absence of LIF. Inclusion of LIF maintained 100% survival of plated neurons over 42 h of culture; without LIF, a large fraction of the neurons did not survive. LIF appeared to maintain survival by preferentially preserving a population of bipolar neurons, while having little effect on the number of monopolar neurons. This work provides quantitative measures of survival and morphology of auditory neurons in vitro. The results support the idea that survival of spiral ganglion neurons in vivo may depend on interactions with adjacent, non-neural tissue and raise the possibility that maintenance of bipolar morphology after hair cell damage may require biochemical mechanisms in addition to those induced by neurotrophins.

The identification and localization of a human gene with sequence similarity to Polycomblike of Drosophila melanogaster.

The Drosophila Polycomb group (PcG) of genes is required for the epigenetic regulation of a number of important developmental genes, including the homeotic (Hox) genes. The members of this gene family encode proteins that do not share sequence similarity, implying that each plays a unique role in this epigenetic repression mechanism. Polycomblike (Pcl) was the second PcG gene to be identified. We report here the isolation and characterization of a human cDNA, termed PHF1, which encodes a protein with significant sequence similarity to Drosophila Polycomblike (PCL). The region of similarity between PHF1 and PCL includes the two PHD fingers (C4-H-C3 motif), the region between them, and sequences C-terminal to the PHD fingers. PHF1 and PCL are 34% identical over this 258-residue region. PHF1 was mapped to 6p21.3 by fluorescence in situ hybridization. While several genetic diseases that are likely to result from developmental abnormalities map to this region, PHF1 is not a clear candidate gene for any of them.

Flagellin gene and protein variation amongst clinical isolates of Pseudomonas aeruginosa.

Flagellin gene sequences from 64 clinical isolates of the opportunistic pathogen Pseudomonas aeruginosa were amplified by PCR and subjected to RFLP analysis by using seven restriction enzymes to digest the amplified products. Using this approach the isolates were assigned to one of 13 groups. The method was rapid, reproducible and applicable to all isolates. In contrast, serotyping failed to satisfactorily resolve 49% of the strains tested. The vast majority of clinical isolates generated amplified products of 1.02 kb (type a) or 1.25 kb (type b). Electron microscopical analysis revealed evidence fax some. flagellar structural variation between P. aeruginosa strains. This study provides further evidence that the flagellin gene is a widely applicable and useful genetic marker for studying genetic variation within populations of closely related bacteria.

The stoned locus of Drosophila melanogaster produces a dicistronic transcript and encodes two distinct polypeptides.

The stoned gene of Drosophila melanogaster is required for normal neuronal function in both adult and larva. We have identified DNA sequences that lie within a genetic region that is known to include the stoned gene and that also reveal restriction site variations in two stoned lethal mutants. This genomic region contains a single transcription unit coding for an approximately 8.4-kb transcript. The transcript is preferentially expressed in the head of adult flies. The isolation and sequencing of cDNA and genomic clones reveals that stoned appears to encode a dicistronic mRNA, although the possible existence of other forms of mRNA cannot be excluded. Antibody cross-reactivity shows that two proteins are translated from the stoned locus in vivo. Both open reading frames (ORFs) encode novel proteins. The protein encoded by the first ORF contains four tandemly repeated motifs, and one domain of the protein encoded by the second ORF shows similarity to a family of proteins (AP50s) associated with clathrin assembly protein complexes.

Nurse managers: perceptions of their role.

A qualitative study on the role of the nurse manager was conducted based on interviews with nurse managers in an acute care teaching hospital. Role descriptions revealed many similarities to Mintzberg's (1973) model of management roles, but additional roles, related to the nurse manager's professional responsibilities were identified. Conflicts within and among roles and a strong need for support were themes that were identified throughout the interviews. Suggestions for improving the quality of worklife of the nurse manager and for further research are included.

Further studies on the actions of endothelin-1 on blood flow in human skin.

When injected into human skin, endothelin-1 produces intense vasoconstriction localized to the site of the injection, but this area of vasoconstriction is surrounded by vasodilatation which spreads several centimetres from the injection site. The vasodilatation induced by intradermal injection of endothelin-1 (63 pmol) into human skin is prevented by local anaesthetic. Pretreatment of human skin with capsaicin also inhibits this response. Pretreatment of subjects with the selective histamine H1-receptor antagonist cetirizine, 10 mg orally 4 h before intradermal injections, inhibited vasodilatation caused by the intradermal injection of histamine (750 pmol), endothelin-1 (63 pmol), and carbachol (750 pmol). Endothelin-1 (0.3-10 microM) and carbachol (1-30 microM) failed to induce histamine release from rat peritoneal mast cells. We conclude that the vasodilatation caused by intradermal injection of endothelin-1 into human skin is neurogenic and is probably mediated by neuropeptide-containing primary afferent neurones. Because neither carbachol nor endothelin-1 cause histamine release from mast cells, our data suggest that histamine release from mast cells at the effector end of the axon reflex is responsible for the carbachol- and endothelin-induced vasodilatation in human skin.

Comparison of infrared and helium-neon lasers in the measurement of blood flow in human skin by the laser-Doppler technique.

Histamine-induced changes in blood flow in normal human skin were assessed by laser-Doppler velocimetry using 2 lasers of different wavelengths: 780 nm infrared and 633 nm helium-neon. The visible flare response in skin was also measured by planimetry. Laser-Doppler velocimetry was shown to detect both the magnitude of the change in blood flow caused by histamine and also the extent of the changes in the skin. Both parameters were related to the dose of histamine, which ranged between 25 and 750 pmol/site. There was good correlation between the magnitude of the histamine-induced blood flow change and the extent of the response. The flare induced by histamine and measured by planimetry was similar in extent to the blood flow change recorded by laser-Doppler velocimetry. No difference in either the magnitude or the extent of blood flow changes in response to histamine, as measured by laser-Doppler velocimetry, were found when lasers of different wavelengths were used.

Squamous cell papilloma at the tracheoesophageal puncture stoma.

Over 140 tracheoesophageal punctures were performed at the Cleveland Clinic Foundation, Department of Otolaryngology and Communicative Disorders over the last seven years. In all cases the Blom-Singer duckbill prosthesis was utilized for alaryngeal voice restoration. Of these patients, two examples were noted where squamous cell papilloma was found at the tracheoesophageal stoma site. The preoperative evaluation and surgical laser technique for surgical excision of the papilloma with postoperative care are described. The case reports of each of the patients that have been diagnosed as having squamous cell papilloma of the tracheoesophageal puncture site are reviewed.

Coordinated feedback regulation of muscle glycogen metabolism: inhibition of purified phosphorylase phosphatase by glycogen.

An assay method was devised for accurately measuring the activity of phosphorylase phosphatase in the presence of glycogen. Both oyster glycogen and rabbit liver glycogen inhibited the activity of purified phosphorylase phosphatase catalytic subunit in a concentration-dependent manner. In the presence of 120 mM KCl, micromolar AMP increased the sensitivity of the phosphatase to glycogen inhibition. The present studies suggest that glycogen feedback inhibition may be a coordinated mechanism producing a decrease in phosphorylase phosphatase activity as well as glycogen synthase phosphatase activity.

Regulation of the Ascaris suum pyruvate dehydrogenase complex by phosphorylation and dephosphorylation.

The pyruvate dehydrogenase complex isolated from 'anaerobic' mitochondria of Ascaris suum has a subunit composition similar to complexes isolated from most other eukaryotic organisms and is regulated by phosphorylation and dephosphorylation. Pyruvate dehydrogenase kinase activity is stimulated by NADH and a number of physiologically important acyl-CoA intermediates and is inhibited by CoA, propionate, tiglate and pyruvate. It is suggested that the elevated levels of pyruvate observed in the ascarid organelle may be important in maintaining the pyruvate dehydrogenase complex in an active state, even in the presence of a reduced pyridine nucleotide pool.