bluuebunny/crossref_metadata_2025 huggingface dataset:
Imaging Synaptic Protein Dynamics Using Photoactivatable Green Fluorescent Protein
Authors: Weimer Robby M. , Hill Travis C. , Hamilton Andrew M. , Zito Karen
Published: July 2012

Considerable evidence has accumulated that structural changes in dendritic spines and their synapses are associated with adaptive functional changes in cortical circuits, such as during circuit refinement in young animals and in learning and memory in adults. Understanding the mechanisms of circuit plasticity requires detailed investigation of the structural dynamics of dendritic spines and how they are regulated by neural activity and sensory experience. Studying the dynamic localization of synaptic proteins in dendritic spines and how their stabilization and exchange rates influence spine structural plasticity is also important. This protocol describes imaging approaches to study synaptic protein dynamics in dendritic spines of the rodent cerebral cortex. It gives a strategy for generating photoactivatable green fluorescent protein (PA-GFP)-tagged synaptic proteins and in vitro and in vivo transfection methods for coexpression of these proteins with a spectrally separable cell-filling marker (DsRed-Express). Methods for tracking synaptic protein localization using photoactivation and time-lapse imaging of PA-GFP in spiny pyramidal neuron dendrites are given. A discussion of imaging hardware and software preferences is also included. The methods described here can be used to study the dynamic processes underlying spine synapse development during the formation and plasticity of neural circuits in the mammalian brain.
Topical Application of Indicators for Calcium Imaging at the Drosophila Larval Neuromuscular Junction
Authors: Macleod Gregory T.
Published: July 2012

Calcium imaging is a technique in which Ca2+-binding molecules are loaded into live cells and as they bind Ca2+ they “indicate” the concentration of free calcium through a change in either the intensity or the wavelength of light emitted (fluorescence or bioluminescence). There are several possible methods for loading synthetic Ca2+ indicators into subcellular compartments, including topical application of membrane-permeant Ca2+ indicators, forward-filling of dextran conjugates, and direct injection. Calcium imaging is a highly informative technique in neurobiology because Ca2+ is involved in many neuronal signaling pathways and serves as the trigger for neurotransmitter release. This article describes the topical application of Ca2+ indicators at the Drosophila larval neuromuscular junction (NMJ). This loading technique is simple to execute and yields data quickly. The drawback is that the data can be difficult to interpret, primarily because it is difficult to ascertain which cellular and subcellular compartment(s) are loaded (e.g., muscle, nerve, or glia; cytosol, mitochondrion, or endoplasmic reticulum).
Dimerizer-Mediated Regulation of Gene Expression In Vitro
Authors: Rivera Victor M. , Berk Lori , Clackson Tim
Published: July 2012

Several systems have been developed that allow transcription of a target gene to be chemically controlled, usually by an allosteric modulator of transcription factor activity. An alternative is to use chemical inducers of dimerization, or “dimerizers,” to reconstitute active transcription factors from inactive fusion proteins. The most widely used system employs the natural product rapamycin, or a biologically inert analog, as the dimerizing drug. A key feature of this system is the tightness of regulation, with basal expression usually undetectable and induced expression levels comparable to constitutive promoters. In our experiments, the use of the minimal interleukin-2 (IL-2) promoter is an important determinant of this; substitution of a minimal simian virus 40 (SV40) or cytomegalovirus (CMV) promoter results in significantly higher levels of basal expression. The key factor dictating the successful use of the system is achieving high expression levels of the activation domain fusion protein. In the context of clinical gene therapies, the system has the advantage of being built exclusively from human proteins, potentially minimizing immunogenicity in the clinical setting. The dimerizer system has been successfully incorporated into diverse vector backgrounds and has been used to achieve long-term regulated gene expression in vitro and in vivo. This protocol describes the preparation of vectors for rapamycin- or rapalog-inducible gene expression, followed by induction of gene expression in vitro.
Generation of Recombinant Alphaviral Vectors
Authors: Lundstrom Kenneth
Published: July 2012

The alphaviruses Semliki Forest virus and Sindbis virus have been used frequently as expression vectors in vitro and in vivo. Usually, these systems consist of replication-deficient vectors that require a helper vector for packaging of recombinant particles. Replication-proficient vectors have also been engineered. Alphaviral vectors can be used as nucleic-acid-based vectors (DNA and RNA) or infectious particles. High-titer viral production is achieved in <2 d. The broad host range of alphaviruses facilitates studies in mammalian and nonmammalian cell lines, primary cells in culture, and in vivo. The strong preference for expression in neuronal cells has made alphaviruses particularly useful in neurobiological studies. Unfortunately, their strong cytotoxic effect on host cells, relatively short-term transient expression patterns, and the reasonably high cost of viral production remain drawbacks. However, novel mutant alphaviruses have showed reduced cytotoxicity and prolonged expression. Membrane proteins (which are generally difficult to express at high levels in recombinant systems) have generated high yields and facilitate applications in structural biology. Alphaviruses have also been applied in vaccine development and gene therapy. This protocol describes the production of recombinant alphaviral vectors. The SFV- and SIN-based expression systems apply two vectors for recombinant particle production. In addition to the RNA-based vectors described here, DNA vectors with cytomegalovirus or other RNA polymerase type II promoters can be used for direct plasmid DNA transfections. Cotransfection of SFV-based pSCA expression and pSCA helper vectors generates recombinant viral particles.
Purification and Concentration of Alphavirus
Authors: Lundstrom Kenneth
Published: July 2012

The alphaviruses Semliki Forest virus and Sindbis virus have been used frequently as expression vectors in vitro and in vivo. Usually, these systems consist of replication-deficient vectors that require a helper vector for packaging of recombinant particles. Replication-proficient vectors have also been engineered. Alphaviral vectors can be used as nucleic-acid-based vectors (DNA and RNA) or infectious particles. High-titer viral production is achieved in <2 d. The broad host range of alphaviruses facilitates studies in mammalian and nonmammalian cell lines, primary cells in culture, and in vivo. The strong preference for expression in neuronal cells has made alphaviruses particularly useful in neurobiological studies. Unfortunately, their strong cytotoxic effect on host cells, relatively short-term transient expression patterns, and the reasonably high cost of viral production remain drawbacks. However, novel mutant alphaviruses have showed reduced cytotoxicity and prolonged expression. Membrane proteins (which are generally difficult to express at high levels in recombinant systems) have generated high yields and facilitate applications in structural biology. Alphaviruses have also been applied in vaccine development and gene therapy. Generally, purification or concentration of alphaviruses is not necessary. However, for instance, the medium derived from baby hamster kidney cells is toxic to primary neurons in culture. Including a purification step substantially improves the survival of the transduced neurons. Viral concentration and purification may also be advantageous for in vivo studies in animal models and are mandatory for clinical applications. This protocol describes three methods for purification and concentration of alphavirus.
Calcium Imaging at the Drosophila Larval Neuromuscular Junction
Authors: Macleod Gregory T.
Published: July 2012

Calcium imaging uses optical imaging techniques to measure the concentration of free calcium [Ca2+] in live cells. It is a highly informative technique in neurobiology because Ca2+ is involved in many neuronal signaling pathways and serves as the trigger for neurotransmitter release. The technique relies on loading Ca2+ indicators into cells, measuring the quantity and/or wavelength of the photons emitted by the Ca2+ indicator, and interpreting these data in terms of [Ca2+]. There are several possible methods for loading synthetic Ca2+ indicators into subcellular compartments, for example, topical application of membrane-permeant Ca2+ indicators, forward-filling of dextran conjugates, and direct injection. These techniques are applicable to calcium imaging at the Drosophila larval neuromuscular junction (NMJ), and are also readily adaptable to Drosophila embryo and adult preparations.
The in vitro blood‐O2 affinity of triploid rainbow trout Oncorhynchus mykiss at different temperatures and CO2 tensions
Authors: Verhille C. , Farrell A. P.
Published: July 2012

Blood‐O2 affinity, Hill number and predicted arterial‐O2 saturation did not differ between diploid (2N) and triploid (3N) rainbow trout Oncorhynchus mykiss blood when compared under various temperature and CO2 partial pressure combinations. These results suggest that reduced hypoxia and warm‐water tolerance and aerobic capacity of 3N fishes are not due to altered blood‐O2 affinity. Further investigation into O2 transport–diffusion capacity and intracellular pH within 3N fishes may further unravel the mechanisms behind reduced 3N tolerance of suboptimal environments.
Dreamlet source‐receiver survey sinking prestack depth migration
Authors: Wu Bangyu , Wu Ru‐shan , Gao Jinghuai
Published: July 2012

Survey sinking migration downward continues the entire surface observed multi‐shot data to the subsurface step by step recursively. Reflected energy from reflectors at current depth appear at zero time and zero offset in the extrapolated wavefield. The data (seismic records) of t > 0 at this depth are equivalent to the data acquired by a survey system deployed at this depth. This is the reason to name the process ‘survey sinking’. The records of negative time need not to be further propagated since they carry no information to image structures beneath the new survey system. In this paper, we combine survey sinking with dreamlet migration. The dreamlet migration method decomposes the seismic wavefield and one‐way wave propagator by complete time‐space localized bases. The localization on time gives flexibility on time‐varying operations during depth extrapolation. In dreamlet survey sinking migration, it only keeps the data for imaging the structures beneath the sunk survey system and gets rid of the data already used to image structures above it. The deeper the depth is, the shorter is the valid time records of the remaining data and less computation is needed for one depth step continuation. For data decomposition, in addition to time axis, dreamlet survey sinking also decomposes the data for source and receiver gathers, which is a fully localized decomposition of prestack seismic data. A three‐scatter model is first used to demonstrate the computational feature and principle of this method. Tests on the two‐dimensional SEG/EAGE salt model show that with reduced data sets the proposed method can still obtain good imaging quality on complex geology structures and a strong velocity contrast environment.
Persistence of complex vascular lesions despite prolonged prostacyclin therapy of pulmonary arterial hypertension
Authors: Pogoriler Jennifer E , Rich Stuart , Archer Stephen L , Husain Aliya N
Published: July 2012

Pogoriler J E, Rich S, Archer S L & Husain A N  (2012) Histopathology 61, 597–609

Persistence of complex vascular lesions despite prolonged prostacyclin therapy of pulmonary arterial hypertension

Aims:  Continuous infusion of prostacyclin analogues improves survival in advanced pulmonary arterial hypertension. In addition to its vasodilatory effects, prostacyclin has the potential to decrease inflammation, thrombosis, and smooth muscle proliferation. The aim of this retrospective study was to determine whether pathological data support the ability of prostanoids to prevent progression of vascular disease.

Methods and results:  Twenty‐two autopsied patients with World Health Organization category 1 pulmonary arterial hypertension (primarily idiopathic and connective tissue disease‐associated) were divided into those who received long‐term prostacyclin (n = 12, PG‐long, mean treatment 3.9 years) and those who received 0–1 month of prostacyclin (n = 10, PG‐short). Surprisingly, PG‐long patients had larger plexiform lesions (P < 0.05), with no decrease in medial and intimal thicknesses as compared with PG‐short patients. Plexiform lesion size and density increased with increasing treatment time. Also, PG‐long patients had fewer platelet thrombi and more frequent acute diffuse alveolar haemorrhage. Quantification of macrophages and T cells revealed no differences in inflammatory infiltrates.

Conclusions:  Although long‐term prostacyclin therapy may have an antithrombotic effect in addition to its vasodilatory actions, it was not associated with the prevention of advanced vascular lesions. The mechanism by which prostacyclin analogues improve survival in pulmonary arterial hypertension remains uncertain.
Loss in CD4 T‐cell responses to multiple epitopes in influenza due to expression of one additional MHC class II molecule in the host
Authors: Nayak Jennifer L. , Sant Andrea J.
Published: July 2012

An understanding of factors controlling CD4 T‐cell immunodominance is needed to pursue CD4 T‐cell epitope‐driven vaccine design, yet our understanding of this in humans is limited by the complexity of potential MHC class II molecule expression. In the studies described here, we took advantage of genetically restricted, well‐defined mouse strains to better understand the effect of increasing MHC class II molecule diversity on the CD4 T‐cell repertoire and the resulting anti‐influenza immunodominance hierarchy. Interferon‐γ ELISPOT assays were implemented to directly quantify CD4 T‐cell responses to I‐Ab and I‐As restricted peptide epitopes following primary influenza virus infection in parental and F1 hybrid strains. We found striking and asymmetric declines in the magnitude of many peptide‐specific responses in F1 animals. These declines could not be accounted for by the lower surface density of MHC class II on the cell or by antigen‐presenting cells failing to stimulate T cells with lower avidity T‐cell receptors. Given the large diversity of MHC class II expressed in humans, these findings have important implications for the rational design of peptide‐based vaccines that are based on the premise that CD4 T‐cell epitope specificity can be predicted by a simple cataloguing of an individual’s MHC class II genotype.
Perceived treatment group affects behavioral and neural responses to visceral pain in a deceptive placebo study
Authors: Kotsis V. , Benson S. , Bingel U. , Forsting M. , Schedlowski M. , Gizewski E. R. , Elsenbruch S.
Published: July 2012

Background  To assess effects of perceived treatment (i.e. drug vs placebo) on behavioral and neural responses to rectal pain stimuli delivered in a deceptive placebo condition.

Methods  This fMRI study analyzed the behavioral and neural responses during expectation‐mediated placebo analgesia in a rectal pain model. In N = 36 healthy subjects, the blood oxygen level‐dependent (BOLD) response during cued anticipation and painful stimulation was measured after participants were informed that they had a 50% chance of receiving either a potent analgesic drug or an inert substance (i.e., double‐blind administration). In reality, all received placebo. We compared responses in subjects who retrospectively indicated that they received the drug and those who believed to have received placebo.

Key Results  55.6% (N = 20) of subjects believed that they had received a placebo, whereas 36.1% (N = 13) believed that they had received a potent analgesic drug. Subjects who were uncertain (8.3%, N = 3) were excluded. Rectal pain‐induced discomfort was significantly lower in the perceived drug treatment group (P < 0.05), along with significantly reduced activation of the insular, the posterior and anterior cingulate cortices during pain anticipation, and of the anterior cingulate cortex during pain (all P < 0.05 in regions‐of‐interest analyses).

Conclusions & Inferences  Perceived treatment constitutes an important aspect in placebo analgesia. A more refined understanding of individual treatment expectations and perceived treatment allocation has multiple implications for the design and interpretation of clinical trials and experimental studies on placebo and nocebo effects.
Using Mixture Models with Linear Predictors to Identify Incorrect Gestational Age in State Birth Records
Authors: Leiss Jack K. , Suchindran C. M. , Kruse Lakota
Published: July 2012

Background:  Birthweight distributions for early last‐menstrual‐period‐based gestational ages are bimodal, and some birthweights in the right‐side distribution are implausible for the specified gestational age. Mixture models can be used to identify births in the right‐side distribution. The objective of this study was to determine which maternal and infant factors to include in the mixture models to obtain the best fitting models for New Jersey state birth records.

Methods:  We included covariates in the models as linear predictors of the means of the component distributions and the proportion of births in each component. This allowed both the means and the proportions to vary across levels of the covariates.

Results:  The final model included maternal age and timing of entry into prenatal care. The proportion of births in the right‐side distribution was lowest for older mothers who entered prenatal care early, higher for teen mothers who entered prenatal care early, higher still for older mothers who entered prenatal care late, and highest for teens who entered prenatal care late. Over 44% of births were classified as incorrect reported gestational age.

Conclusion:  These results suggest that (1) including these two covariates as linear predictors of the means and mixing proportions gives the best model for identifying births with incorrect reported gestational age, (2) late entry into prenatal care is a mechanism by which erroneously short last‐menstrual‐period‐based gestational ages are generated, and (3) including linear predictors of the mixing proportions in the model increases the validity of the classification of incorrect reported gestational age.
Development and validation of standard area diagrams to aid assessment of pecan scab symptoms on fruit
Authors: Yadav N. V. S. , de Vos S. M. , Bock C. H. , Wood B. W.
Published: July 2012

Pecan scab (Fusicladium effusum) causes losses of pecan nutmeat yield and quality in the southeastern United States. Disease assessment relies on visual rating, which can be inaccurate and imprecise, with poor inter‐rater reliability. A standard area diagram (SAD) set for pecan scab on fruit valves was developed. A set of 40 images of diseased fruit valves with known severity was assessed twice by 23 raters. The first assessment was conducted without SADs, and the second assessment was made using the SADs as an aid. SADs improved rater accuracy (correction factor, Cb = 0·86 and 0·97, without and with SADs, respectively) and agreement (Lin’s concordance correlation coefficient, ρc = 0·79 and 0·89 without and with SADs, respectively) with true values. SADs improved inter‐rater reliability (intra‐class correlation coefficient, ρ = 0·77 and 0·96 without and with SADs, respectively). The least accurate and precise raters without SADs improved more using SADs compared to the most accurate and precise raters. Experienced raters had significantly higher accuracy and precision compared to inexperienced raters, but only when unaided by the SAD set. There was no significant difference in time to assess images without SADs, but experienced raters using SADs were faster compared to inexperienced raters. There was a slight tendency for faster raters to assess more slowly, and slower raters to assess faster when using SADs. SADs improve rater estimates of pecan scab severity on fruit, and this SAD set should be useful for assessment where greater precision, accuracy and inter‐rater reliability are required.
Generation of pathogenic F1 progeny from crosses of Phytophthora infestans isolates differing in ploidy
Authors: Hamed B. H. , Gisi U.
Published: July 2012

Ten Phytophthora infestans isolates were analysed for maximal growth rate (μmax), sporulation capacity (K), ploidy and the ability to produce infective F1 progeny when combined in different crosses on tomato leaflets. Ploidy was determined using three different methods: simple sequence repeats (SSR, microsatellites), allele quantification at F382Y in the RNA polymerase, and flow cytometry. Eight out of 10 isolates were classified as diploid, the other two as triploid. There was no correlation between ploidy, growth rate and sporulation capacity although the latter was rather low for triploid isolates. Ten crosses were investigated in tomato leaflets (five 2n × 2n; four 2n × 3n; one 3n × 3n). Oospore production was observed in all crosses independent of parental ploidy. Germination and pathogenicity of oospores were investigated by measuring the number of infection sites (pathogenic F1 progeny) over a time period of 72 days. All crosses containing a triploid parent produced many fewer F1 isolates than 2n × 2n crosses. The generation of pathogenic F1 progeny isolates was best for 2n × 2n crosses. A total of 319 F1 offspring were produced and analysed for mating type and ploidy; a subset was tested for ploidy with all three methods. For 2n × 2n crosses, the majority of offspring were diploid, whereas in crosses containing one or two triploid parents, diploid, triploid and trisomic offspring were observed. The results suggest that less pathogenic F1 progeny isolates are produced if at least one parent is triploid.
Extended‐release naltrexone modulates brain response to drug cues in abstinent heroin‐dependent patients
Authors: Langleben Daniel D. , Ruparel Kosha , Elman Igor , Loughead James W. , Busch Elliot L. , Cornish James , Lynch Kevin G. , Nuwayser Elie S. , Childress Anna R. , O'Brien Charles P.
Published: July 2012

Drug cues play an important role in relapse to drug use. Naltrexone is an opioid antagonist that is used to prevent relapse in opioid dependence. Central opioidergic pathways may be implicated in the heightened drug cue‐reactivity, but the effects of the opioid receptors' blockade on the brain responses to drug cues in opioid dependence are unknown. To pursue this question, we studied 17 abstinent i.v. heroin users with brain functional magnetic resonance imaging (fMRI) during exposure to visual heroin‐related cues and matched neutral images before and 10–14 days after an injection of extended‐release naltrexone (XRNTX). Whole brain analysis of variance of fMRI data showed main effect of XRNTX in the medial frontal gyrus, precentral gyrus, cuneus, precuneus, caudate and the amygdala. fMRI response was decreased in the amygdala, cuneus, caudate and the precentral gyrus and increased in the medial frontal gyrus and the precuneus. Higher plasma levels of naltrexone's major metabolite, 6‐beta‐naltrexol, were associated with larger reduction in the fMRI response to drug cues after XRNTX in the precentral, caudate and amygdala clusters. The present data suggest that XRNTX pharmacotherapy of opioid‐dependent patients may, respectively, decrease and potentiate prefrontal and limbic cortical responses to drug cues and that this effect might be related to the XRNTX metabolism. Our findings call for further evaluation of the brain fMRI response to drug‐related cues and of the 6‐beta‐naltrexol levels as potential biomarkers of XRNTX therapeutic effects in patients with opioid dependence.
Effects of pre‐weaning dietary substitutions on plasma insulin and glucose profiles in primiparous sows
Authors: Chen T. Y. , Stott P. , O’Leary S. , Athorn R. Z. , Bouwman E. G. , Langendijk P.
Published: July 2012

The objective of this study was to investigate the effects of substituting 1 kg of a standard lactation diet with 1 kg of a sugar‐rich (15.75 DE MJ/kg) or fat‐rich (23.85 DE MJ/kg) diet during late lactation on blood glucose and insulin changes in primiparous sows. During a 4‐week lactation period, 21 primiparous sows were fed to appetite with a standard lactation diet (14.10 DE MJ/kg). At 9 days before weaning, sows were assigned to a control (C, n = 7), fat (F, n = 6) or sugar (S, n = 8) treatment. During the treatment period (from 8 days before weaning until weaning), 1 kg of the lactation diet was substituted with 1 kg of a sugar‐rich or fat‐rich diet for S and F sows. At 3 days before weaning, serial blood samples were collected for a total of 228 min around feeding to establish pre‐ and postprandial plasma glucose and insulin concentrations. Preprandial plasma glucose and insulin concentrations did not differ between treatments (p > 0.05); however, mean plasma glucose and insulin concentrations were higher for S compared to F (p < 0.05) and intermediate for the C sows. Postprandial plasma concentrations of glucose and insulin were higher for the S sows than for C and F sows (p < 0.05). Sow body weight loss during late lactation did not differ between treatments (p > 0.05). The results from our study suggest that a sugar‐enriched diet during the last week of lactation elevates circulating glucose and insulin concentrations and may potentially improve post‐weaning fertility in primiparous sows.
Natural Spread of Plant Viruses by Birds
Authors: Peters Dick , Engels Coert , Sarra Soungalo
Published: July 2012

Observations made in Mali strongly suggest that Rice yellow mottle virus (RYMV) is spread by weaverbirds (Quelea quelea) below and around baobab trees (Adansonia digitata) in which they nest. Rice leaves in bird nests appeared to be infected. In Spain, an infection of Southern bean mosaic virus (SBMV) in string (climbing) beans (Phaseolus vulgaris) was apparently introduced and spread by sparrows (Passer domesticus) judging from the damage caused on flowers and bean pods. Damaged leaves and pods on SBMV‐infected plants were also found in a screenhouse visited by sparrows and bulbuls (Pycnonotus barbatus) in Morocco. These observations showed that both viruses could be spread by birds when either collecting infected leaves for nesting or feeding on infected plants.
Canine‐chilled Sperm: Study of a Semen Extender Made with Low‐density Lipoproteins from Hen Egg Yolk Supplemented with Glutamine
Authors: Bencharif D , Amirat‐Briand L , Le Guillou J , Garand A , Anton M , Schmitt E , Desherces S , Delhomme G , Langlois M‐L , Destrumelle S , Vera‐Munoz O , Barrière P , Tainturier D
Published: July 2012

Contents

The objective of this study was to evaluate the effects of a combination of 6% low‐density lipoproteins (LDL) and 20 mm glutamine in comparison with other extenders used for the refrigeration of canine semen: Tris egg yolk (EY) 20% and 6% LDL. The percentages of mobile spermatozoa after 4 days storage in a domestic refrigerator at +4°C were 53.1%, 44.2% and 52.2% for the 6% LDL + 20 mm glutamine, 20% EY and 6% LDL extenders respectively for 100% of the dogs. After 7 days of storage, these percentages fell to 37.8%, 26.4% and 33.6% in the same extenders for 50% of the dogs. In vitro fertility tests were performed with all of the extenders following the mobility results. These tests were conducted on the day of sampling (D0), and 48 and 96 h after sampling. The results of the hypo‐osmotic swelling test were 82.6%, 81.2% and 85.7% on D0, 75.2%, 74.1% and 78.5% on D2, and 70.8%, 71% and 76.1% on D4 for the 6% LDL + 20 mm glutamine, 20% EY and 6% LDL extenders, respectively. For the FITC/pisum sativum agglutinin (PSA) test, the results were 81.5%, 70.2% and 84.8% on D0, 78.9%, 62.3% and 84.2% on D2, and 72.7%, 59.6% and 73.7% on D4 for the 6% LDL + 20 mm glutamine, 20% EY and 6% LDL extenders, respectively. The acridine orange test was positive; in nearly 100% of cases, none of the spermatozoa had been denatured on D0, D2 and D4. The 6% LDL + 20 mm glutamine and the 6% LDL extenders are capable of preserving spermatozoa that have been stored in a domestic refrigerator at +4°C for at least 4 days. This means that the spermatozoa retain good cytoplasmic membrane integrity, had not capacitated and contained intact DNA in comparison with spermatozoa preserved in the egg yolk extender. The duration of storage is a very important consideration when faced with the problem of sending semen over ever‐greater distances.
Prognostic factors for treatment outcomes in transient tachypnea of the newborn
Authors: Bak Seon‐Yeong , Shin Youn‐Ho , Jeon Ji‐Hyun , Park Kyu‐Hee , Kang Jin‐Hee , Cha Dong‐Hyun , Han Man‐Yong , Jo Heui‐Seung , Lee Kyu‐Hyung , Lee Cho‐Ae
Published: July 2012

Background:  Transient tachypnea of the newborn (TTN) is a clinical syndrome associated with respiratory distress usually seen shortly after delivery in infants. This study aims to determine the risk factors predicting treatment outcomes in infants with TTN.

Methods:  Data from 236 infants diagnosed with TTN during the study period were evaluated retrospectively. Logistic regression analyses were performed to select significant risk factor for prognosis (prolonged oxygen therapy, application of mechanical ventilator, and prolonged hospital stay) of TTN among components of clinical variables.

Results:  Of the 236 TTN infants, 111 (47.0%) infants were delivered via cesarean section (CS) without labor, 29 (12.3%) infants were delivered via CS with labor, and 96 (40.7%) were delivered via vaginal birth. Lower Apgar score at 1 min (OR: 3.03; 95%CI: 1.25–7.36) and lower umbilical artery pH (OR: 4.00; 95%CI 1.55–10.49) were associated with a significantly increased risk for mechanical ventilator care. Also, late‐preterm delivery (OR: 4.70; 95%CI: 2.11–10.49) was independently associated with risk of prolonged duration of hospital stay.

Conclusions:  Late‐preterm delivery, lower initial umbilical artery pH (<7.25), and lower Apgar score at 1 min were independently associated with poor prognostic treatment outcomes in infants with TTN.
Resonance properties of different neuronal populations in the immature mouse neocortex
Authors: Sun Haiyan , Luhmann Heiko J. , Kilb Werner
Published: July 2012

In vivo recordings in the immature neocortex revealed spontaneous and sensory‐driven oscillatory activity from delta (0.5–4 Hz) to gamma (30–100 Hz) frequencies. In order to investigate whether the resonance properties of distinct neuronal populations in the immature neocortex contribute to these network oscillations, we performed whole‐cell patch‐clamp recordings from visually identified neurons in tangential and coronal neocortical slices from postnatal day (P)0–P7 C57Bl/6 mice. Subthreshold resonance was analysed by sinusoidal current injection of varying frequency. All Cajal–Retzius cells showed subthreshold resonance, with an average frequency of 2.6 ± 0.1 Hz (n = 60), which was massively reduced by ZD7288, a blocker of hyperpolarization‐activated cation currents. Approximately 65.6% (n = 61) of the supragranular pyramidal neurons showed subthreshold resonance, with an average frequency of 1.4 ± 0.1 Hz (n = 40). Application of Ni2+ suppressed subthreshold resonance, suggesting that low‐threshold calcium currents contribute to resonance in these neurons. Approximately 63.6% (n = 77) of the layer V pyramidal neurons showed subthreshold resonance, with an average frequency of 1.4 ± 0.2 Hz (n = 49), which was abolished by ZD7288. Only 44.1% (n = 59) of the subplate neurons showed subthreshold resonance, with an average frequency of 1.3 ± 0.2 Hz (n = 26) and a small resonance strength. In summary, these results demonstrate that neurons in all investigated layers show resonance behavior, with either hyperpolarization‐activated cation or low‐threshold calcium currents contributing to the subthreshold resonance. The observed resonance frequencies are in the range of slow activity patterns observed in the immature neocortex, suggesting that subthreshold resonance may support the generation of this activity.
‘Candidatus Halomonas phosphatis’, a novel polyphosphate‐accumulating organism in full‐scale enhanced biological phosphorus removal plants
Authors: Nguyen Hien Thi Thu , Nielsen Jeppe Lund , Nielsen Per Halkjær
Published: July 2012

Microautoradiography combined with fluorescence in situ hybridization (MAR‐FISH) was used to screen for potential polyphosphate‐accumulating organisms (PAOs) in full‐scale enhanced biological phosphorus removal (EBPR) plants. Clone library analyses and application of MAR‐FISH using newly designed probes revealed that small rods related to uncultured Halomonas within the gammaproteobacterial family Halomonadaceae were actively involved in uptake of orthophosphate. Although deeply branched in the Gammaproteobacteria, they were not targeted by the gammaproteobacterial probe (GAM42a). A part of them were also not targeted with the general bacterial probes (EUBmix). They could take up short‐chain fatty acids (e.g. acetate and propionate) and ethanol under both anaerobic and aerobic conditions. Polyhydroxyalkanoate storage was observed under anaerobic conditions. There was no indication of a denitrifying capability. A survey of the occurrence of these Halomonas‐PAOs in 23 full‐scale EBPR plants revealed that they made up 0.5–5.7% of all bacteria in the plants, and were often in higher abundance than the well‐described PAOs ‘Candidatus Accumulibacter phosphatis’. This indicates a potentially important role for these uncultured Halomonas bacteria in the EBPR process in full‐scale plants and we propose to name them ‘Candidatus Halomonas phosphatis’.
PARENTAL SUBSTANCE USE AND FOSTER CARE: EVIDENCE FROM TWO METHAMPHETAMINE SUPPLY SHOCKS
Authors: CUNNINGHAM SCOTT , FINLAY KEITH
Published: July 2012

Foster care caseloads have nearly doubled over the last three decades. Parental methamphetamine (meth) use grew significantly during the same period. While child welfare workers and law enforcement claim that parental meth use contributes to foster care growth, the evidence for a causal effect has not been determined. This paper presents the first evidence of a causal effect of meth on foster care admissions using two exogenous supply‐side interventions in meth markets from the late 1990s for identification. First, we find that restrictions on meth precursor distribution caused meth use (proxied by white meth self‐referred treatment cases) to decline 4.1%. Second, using two‐stage least squares, we estimate a positive elasticity of foster care cases with respect to meth use of 1.54. We also estimate elasticities of 1.03 and 1.49 for cases of child neglect and parental abuse, respectively. These results suggest that child welfare policies should be designed specifically for the children of meth‐using parents. (JEL I12, J13, K42)
UNIONS' RELATIVE CONCERNS AND STRIKES IN WAGE BARGAINING
Authors: Mauleon A. , Vannetelbosch Vincent , Vergari Cecilia
Published: July 2012

We consider a model of wage determination with private information in an oligopoly. We investigate the effects of unions having relative concerns on the negotiated wage and the strike activity. We show that an increase of unions’ relative concerns has an ambiguous effect on the strike activity.
Banks, Financial Markets and International Consumption Risk Sharing
Authors: Leibrecht Markus , Scharler Johann
Published: July 2012

In this article, we explore how characteristics of the domestic financial system influence the international allocation of consumption risk in a sample of OECD countries. Our results show that the extent of risk sharing achieved does not depend on the overall development of the domestic financial system per se. Rather, it depends on how the financial system is organized. Countries characterized by developed financial markets are less exposed to idiosyncratic risk, whereas the development of the banking sector contributes little to the international diversification of consumption risk.
Market Liberalization, Regulatory Uncertainty, and Firm Investment
Authors: Baumann Florian , Friehe Tim
Published: July 2012

This paper analyzes the effects of regulatory uncertainty regarding labor costs on investment in a liberalized market. We distinguish between the external investment margin (market entry) and the internal investment margin (technology), and establish that regulatory uncertainty affects these margins differently, encouraging market entry, but discouraging technological investment. As a consequence, the impact of regulatory uncertainty on competition in liberalized markets is a combination of these two countervailing forces.
Al Rawi, Tariq, and the Future of Closed Material Procedures and Special Advocates
Authors: Ip John
Published: July 2012

This note considers the impact of the Supreme Court's decisions in Al Rawi v The Security Service and Home Office v Tariq on the use of closed material procedures and special advocates. The government's subsequent Justice and Security Green Paper is also discussed.
Independent Schools Council v Charity Commission for England and Wales [2011] UKUT 421 (TCC)
Authors: Synge Mary
Published: July 2012

This note provides a critical analysis of the Upper Tribunal's decision and questions its proposed application and legal justification. The author suggests that the Upper Tribunal has introduced a third sense of public benefit and that this relies upon a circular rationale which is informed by policy rather than law.
A rights approach to supporting the sexual fetish of a man with learning disability: method, process and applied learning
Authors: Cambridge Paul
Published: July 2012

Accessible summary

Some men with learning disabilities have a sexual fetish which may present a support challenge for staff or carers.

Sexual fetish in people with learning disabilities is often ignored, seen as pathological or associated with risk.

It is possible to support sexual fetish in person‐centred ways which respect individual rights and informed choice.

This paper reports on a psycho‐educational intervention associated with the sexual fetish of a man with mild learning disability and autism which centred on his use of nappies and baby paraphernalia. It outlines the nature and expression of his sexual fetish and the risks perceived to be associated with it and describes the approach developed to support him, including the aims which underpinned the work and the methods and processes employed. It was found that a person‐centred psycho‐educational approach from a rights based perspective achieved important changes in his life and self‐esteem and helped challenge assumptions about his sexual risk. The paper also reflects on the organisational location of this work and identifies the applied learning for supporting the sexual fetish of people with learning disabilities, of value for informing sex education and sexuality support for people with learning disabilities more widely.
Expression of cyclooxygenase‐1 (COX‐1) and COX‐2 in human male gametes from normal patients, and those with varicocele and diabetes: a potential molecular marker for diagnosing male infertility disorders
Authors: Perrotta I. , Santoro M. , Guido C. , Avena P. , Tripepi S. , De Amicis F. , Gervasi M. C. , Aquila S.
Published: July 2012

Rising rates of varicocele and diabetes mellitus (DM) pose a significant problem to human fertility. Recent studies have pointed out the impact of cyclooxygenase (COX) in the regulation of testicular function and male fertility. Prominent COX‐2 expression has been described recently in the testes of infertile patients, but little is known about the role and identity of COX isoforms in human sperm under certain disease states such as varicocele and DM. We therefore examined the expression profile and ultrastructural localization of COX‐1 and COX‐2 concomitantly in semen samples from healthy donors, and patients with varicocele and DM. Using Western blotting assay, ‘varicocele’ and ‘diabetic’ sperm showed enhanced COX isoforms expression with respect to the ‘healthy’ sperm. Immunogold labeling revealed human sperm anatomical regions containing COX‐1 and COX‐2, confirming their increased expression in pathological samples. Our data demonstrate that both COX isoforms are upregulated in the spermatozoa of varicocele and diabetic patients, suggesting the harmful effect of the diseases also at the sperm molecular level, going beyond the abnormal morphology described to date. In conclusion, COX enzymes may possess a biological relevance in the pathogenesis and/or maintenance of male factor infertility associated with varicocele and DM, and may be considered additional molecular markers for the diagnosis of male infertility disorders.
Current issues with standards in the measurement and documentation of human skeletal anatomy
Authors: Magee Justin , McClelland Brian , Winder John
Published: July 2012

Digital modeling of human anatomy has become increasingly important and relies on well‐documented quantitative anatomy literature. This type of documentation is common for the spine and pelvis; however, significant issues exist due to the lack of standardization in measurement and technique. Existing literature on quantitative anatomy for the spine and pelvis of white adults (aged 18–65 years, separated into decadal categories) was reviewed from the disciplines of anatomy, manipulative therapy, anthropometrics, occupational ergonomics, biomechanics and forensic science. The data were unified into a single normative model of the sub‐axial spine. Two‐dimensional orthographic drawings were produced from the 590 individual measurements identified, which informed the development of a 3D digital model. A similar review of full range of motion data was conducted as a meta‐analysis and the results were applied to the existing model, providing an inter‐connected, articulated digital spine. During these data analysis processes several inconsistencies were observed accompanied by an evidential lack of standardization with measurement and recording of data. These have been categorized as: anatomical terminology; scaling of measurements; measurement methodology, dimension and anatomical reference positions; global coordinate systems. There is inconsistency in anatomical terminology where independent researchers use the same terms to describe different aspects of anatomy or different terms for the same anatomy. Published standards exist for measurement methods of the human body regarding spatial interaction, anthropometric databases, automotive applications, clothing industries and for computer manikins, but none exists for skeletal anatomy. Presentation of measurements often lacks formal structure in clinical publications, seldom providing geometric reference points, therefore making digital reconstruction difficult. Published quantitative data does not follow existing international published standards relating to engineering drawing and visual communication. Large variations are also evident in standards or guidelines used for global coordinate systems across biomechanics, ergonomics, software systems and 3D software applications. This paper identifies where established good practice exists and suggests additional recommendations, informing an improved communication protocol, to assist reconstruction of skeletal anatomy using 3D digital modeling.
Near‐complete adaptation of the PRiMA knockout to the lack of central acetylcholinesterase
Authors: Farar Vladimir , Mohr Franziska , Legrand Marie , Lamotte d’Incamps Boris , Cendelin Jan , Leroy Jacqueline , Abitbol Marc , Bernard Veronique , Baud Frédéric , Fournet Vincent , Houze Pascal , Klein Jochen , Plaud Benoit , Tuma Jan , Zimmermann Martina , Ascher Philippe , Hrabovska Anna , Myslivecek Jaromir , Krejci Eric
Published: July 2012

J. Neurochem. (2012) 122, 1065–1080.

Acetylcholinesterase (AChE) rapidly hydrolyzes acetylcholine. At the neuromuscular junction, AChE is mainly anchored in the extracellular matrix by the collagen Q, whereas in the brain, AChE is tethered by the proline‐rich membrane anchor (PRiMA). The AChE‐deficient mice, in which AChE has been deleted from all tissues, have severe handicaps. Surprisingly, PRiMA KO mice in which AChE is mostly eliminated from the brain show very few deficits. We now report that most of the changes observed in the brain of AChE‐deficient mice, and in particular the high levels of ambient extracellular acetylcholine and the massive decrease of muscarinic receptors, are also observed in the brain of PRiMA KO. However, the two groups of mutants differ in their responses to AChE inhibitors. Since PRiMA‐KO mice and AChE‐deficient mice have similar low AChE concentrations in the brain but differ in the AChE content of the peripheral nervous system, these results suggest that peripheral nervous system AChE is a major target of AChE inhibitors, and that its absence in AChE‐ deficient mice is the main cause of the slow development and vulnerability of these mice. At the level of the brain, the adaptation to the absence of AChE is nearly complete.
What goes around comes around: Perceived vulnerable employment and economic voting in developing countries
Authors: Singer Matthew M.
Published: July 2012

Voters who perceive the economy to be weak are generally less likely to support the incumbent government. Yet there is a debate over whether all people respond equally to economic shifts or if the state of the economy is more salient for those who feel economically vulnerable. This article examines whether insecure employment situations and employability concerns strengthen responses to the government's economic record. Data from Latin America and Eastern Europe confirm that workers who feel anxious about being fired or who believe it would be difficult to find a new job place significantly greater weight on sociotropic evaluations than do those with more secure employment situations. Thus incumbents who create risks for vulnerable workers are sanctioned, while those who create opportunities are rewarded most.
Nitric oxide potentiation of the homomeric ρ1 GABAC receptor function
Authors: Gasulla J , Beltrán González AN , Calvo DJ
Published: July 2012

BACKGROUND AND PURPOSE NO is a highly diffusible and reactive gas produced in the nervous system, which acts as a neuronal signal mediating physiological or pathological mechanisms. NO can modulate the activity of neurotransmitter receptors and ion channels, including NMDA and GABAA receptors. In the present work, we examined whether GABAC receptor function can also be regulated by NO.

EXPERIMENTAL APPROACH Homomeric ρ1 GABAC receptors were expressed in oocytes and GABA‐evoked responses electrophysiologically recorded in the presence or absence of the NO donor DEA. Chemical protection of cysteines by selective sulfhydryl reagents and site‐directed mutagenesis were used to determine the protein residues involved in the actions of NO.

KEY RESULTS GABAρ1 receptor responses were significantly enhanced in a dose‐dependent, fast and reversible manner by DEA and the specific NO scavenger CPTIO prevented these potentiating effects. The ρ1 subunits contain only three cysteine residues, two extracellular at the Cys‐loop (C177 and C191) and one intracellular (C364). Mutations of C177 and C191 render the ρ1 GABA receptors non‐functional, but C364 can be safely exchanged by alanine (C364A). NEM, N‐ethyl maleimide and (2‐aminoethyl) methanethiosulfonate prevented the effects of DEA on GABAρ1 receptors. Meanwhile, the potentiating effects of DEA on mutant GABAρ1C364A receptors were similar to those observed on wild‐type receptors.

CONCLUSIONS AND IMPLICATIONS Our results suggest that the function of GABAC receptors can be enhanced by NO acting at the extracellular Cys‐loop.