bluuebunny/crossref_metadata_2025_split huggingface dataset:
‘True’ null allele detection in microsatellite loci: a comparison of methods, assessment of difficulties and survey of possible improvements
Authors: Dąbrowski M. J. , Bornelöv S. , Kruczyk M. , Baltzer N. , Komorowski J. 
Published: September 2014

AbstractNull alleles are alleles that for various reasons fail to amplify in a PCR assay. The presence of null alleles in microsatellite data is known to bias the genetic parameter estimates. Thus, efficient detection of null alleles is crucial, but the methods available for indirect null allele detection return inconsistent results. Here, our aim was to compare different methods for null allele detection, to explain their respective performance and to provide improvements. We applied several approaches to identify the ‘true’ null alleles based on the predictions made by five different methods, used either individually or in combination. First, we introduced simulated ‘true’ null alleles into 240 population data sets and applied the methods to measure their success in detecting the simulated null alleles. The single best‐performing method was ML‐NullFreq_frequency. Furthermore, we applied different noise reduction approaches to improve the results. For instance, by combining the results of several methods, we obtained more reliable results than using a single one. Rule‐based classification was applied to identify population properties linked to the false discovery rate. Rules obtained from the classifier described which population genetic estimates and loci characteristics were linked to the success of each method. We have shown that by simulating ‘true’ null alleles into a population data set, we may define a null allele frequency threshold, related to a desired true or false discovery rate. Moreover, using such simulated data sets, the expected null allele homozygote frequency may be estimated independently of the equilibrium state of the population.
Characteristics and outcome of unplanned out‐of‐institution births in Norway from 1999 to 2013: a cross‐sectional study
Authors: Gunnarsson Björn , Smárason Alexander K. , Skogvoll Eirik , Fasting Sigurd 
Published: September 2014

AbstractObjectiveTo study the incidence, maternal characteristics and outcome of unplanned out‐of‐institution births (= unplanned births) in Norway.DesignRegister‐based cross‐sectional study.PopulationAll births in Norway (n = 892 137) from 1999 to 2013 with gestational age ≥22 weeks.MethodsAnalysis of data from the Medical Birth Registry of Norway from 1999 to 2013. Unplanned births (n = 6062) were compared with all other births (reference group).ResultsThe annual incidence rate of unplanned births was 6.8/1000 births and remained stable during the period of study. Young multiparous women residing in remote municipalities were at the highest risk of experiencing unplanned births. The unplanned birth group had higher perinatal mortality rate for the period, 11.4/1000 compared with 4.9/1000 for the reference group (incidence rate ratio 2.31, 95% confidence interval 1.82–2.93, p < 0.001). Annual perinatal mortality rate for unplanned births did not change significantly (p = 0.80) but declined on average by 3% per year in the reference group (p < 0.001). The unplanned birth group had a lower proportion of live births in all birthweight categories. Live born neonates with a birthweight of 750–999 g in the unplanned birth group had a more than five times higher mortality rate during the first week of life, compared with reference births in the same birthweight category.ConclusionsUnplanned births are associated with adverse outcome. Excessive mortality is possibly caused by reduced availability of necessary medical interventions for vulnerable newborns out‐of‐hospital.
Blood pressure, heart rate and tubuloglomerular feedback in A1AR‐deficient mice with different genetic backgrounds
Authors: Kim S. M. , Mizel D. , Qin Y. , Huang Y. , Schnermann J. 
Published: September 2014

AbstractAimDifferences in genetic background between control mice and mice with targeted gene mutations have been recognized as a potential cause for phenotypic differences. In this study, we have used A1AR‐deficient mice in a C57Bl/6 and SWR/J congenic background to assess the influence of background on the effect of A1AR‐deficiency on cardiovascular and renal functional parameters.MethodsIn A1AR+/+ and A1AR−/− mice in C57Bl/6 and SWR/J congenic backgrounds, we assessed blood pressure and heart rate using radio‐telemetry, plasma renin concentrations and tubuloglomerular feedback.ResultsWe did not detect significant differences in arterial blood pressure (MAP) and heart rates (HR) between A1AR+/+ and A1AR−/− mice in either C57Bl/6, SWR/J or mixed backgrounds. MAP and HR were significantly higher in SWR/J than in C57Bl/6 mice. A high NaCl intake increased MAP in A1AR−/− mice on C57Bl/6 background while there was less or no salt sensitivity in the SWR/J background. No significant differences in plasma renin concentration were detected between A1AR−/− and A1AR+/+ mice in any of the strains. Tubuloglomerular feedback was found to be absent in A1AR−/− mice with SWR/J genetic background.ConclusionsWhile this study confirmed important differences between inbred mouse strains, we did not identify phenotypic modifications of A1AR‐related effects on blood pressure, heart rate and plasma renin by differences in genetic background.
The growth‐promoting activity of egg white proteins in the C2C12 myoblast cell line
Authors: Mizunoya Wataru , Tashima Ayumi , Sato Yusuke , Tatsumi Ryuichi , Ikeuchi Yoshihide 
Published: September 2014

AbstractIn this study, we examined the effects of several egg white proteins (ovalbumin, ovomucoid, ovotransferrin and lysozyme) on proliferation and myotube growth in C2C12 murine myoblast cells. Cell proliferation was measured using a water‐soluble tetrazolium salt (WST‐8)‐based assay and then validated using Giemsa staining. Significant proliferative activities of C2C12 cells were observed in response to the addition of 10−5–10−4 mol/L ovalbumin or ovomucoid. Ovotransferrin decreased C2C12 cell proliferation and lysozyme showed no significant effects on the proliferation of C2C12 cells. In contrast, the proliferative effects of ovalbumin and ovomucoid were not observed in 3T3‐L1 murine preadipocyte cells. We also measured the effects of ovalbumin and ovomucoid on C2C12 myotube diameters by using histological analysis. In comparison to control cells, myotube diameters were significantly increased in cells cultured in 10−6–10−4 mol/L ovalbumin or ovomucoid, suggesting that ovalbumin and ovomucoid stimulate the growth of myotubes. Thus, our results clearly demonstrated that ovalbumin or ovomucoid stimulated the proliferation of myoblasts and growth of myotubes.
Knockout of targeted gene in porcine somatic cells using zinc‐finger nuclease
Authors: Hisamatsu Shin , Sakaue Motoharu , Takizawa Akiko , Kato Tsubasa , Kamoshita Maki , Ito Junya , Kashiwazaki Naomi 
Published: September 2014

AbstractTargeted genome editing is a widely applicable approach for efficiently modifying any sequence of interest in animals. It is very difficult to generate knock‐out and knock‐in animals except for mice up to now. Very recently, a method of genome editing using zinc‐finger nucleases (ZFNs) has been developed to produce knockout rats. Since only injection of ZFNs into the pronuclear (PN) embryo is required, it seems to be useful for generating gene‐targeted animals, including domestic species. However, no one has reported the successful production of knockout pigs by direct injection of ZFNs into PN embryos. We examined whether ZFN works on editing the genome of porcine growth hormone receptor in two kinds of cell lines (ST and PT‐K75) derived from the pig as a preliminary study. Our data showed that pZFN1/2 vectors were efficiently transfected into both ST and PT‐K75 cells. In both cell lines, results from Cel‐I assay showed that modification of the targeted gene was confirmed. We injected ZFN1/2 mRNAs into the nucleus of PN stage embryos and then they were transferred to the recipients. However, pups were not delivered. Taken together, ZFN can be an available technology of genome editing even in the pig but further improvement will be required for generating genome‐modified pigs.
Supplementary immunocytochemistry of hepatocyte growth factor production in activated macrophages early in muscle regeneration
Authors: Sawano Shoko , Suzuki Takahiro , Do Mai‐Khoi Q. , Ohtsubo Hideaki , Mizunoya Wataru , Ikeuchi Yoshihide , Tatsumi Ryuichi 
Published: September 2014

AbstractRegenerative intramuscular motor‐innervation is thought to reside in the spatiotemporal expression of axon‐guidance molecules. Our previous studies showed that resident myogenic stem cells, satellite cells, up‐regulate a secreted neural‐chemorepellent semaphorin 3A (Sema3A) during the early‐differentiation period, in response to hepatocyte growth factor (HGF) elevated in injured muscle. However, a paracrine source of the HGF release is still unknown. Very recently, we proposed a possible contribution of anti‐inflammatory macrophages (CD206‐positive M2) by showing that M2 cells infiltrate predominantly at the early‐differentiation phase (3–5 days post‐injury) and produce/secrete large amounts of HGF. However, in understanding this concept there still remains a critical need to examine if phagocytotic pro‐inflammatory macrophages (CD86‐positive M1), another activated‐phenotype still present at the early‐differentiation phase concerned, produce HGF upon muscle injury. The current immunocytochemical study demonstrated that the HGF expression is negative for M1 prepared from cardiotoxin‐injured Tibialis anterior muscle at day 5, in contrast to the intense fluorescent‐signal of M2 served as a positive control. This supplementary result advances our understanding of a spatiotemporal burst of HGF secretion from M2 populations (not M1) to impact Sema3A expression, which ensures a coordinated delay in attachment of motoneuron terminals onto damaged and generating fibers during the early phase of muscle regeneration.
Impact of Alternative Medical Device Approval Processes on Costs and Health
Authors: George Benjamin P. , Venkataraman Vinayak , Dorsey E. Ray , Johnston S. Claiborne 
Published: September 2014

BackgroundMedical devices are often introduced prior to randomized‐trial evidence of efficacy and this slows completion of trials. Alternative regulatory approaches include restricting device use outside of trials prior to trial evidence of efficacy (like the drug approval process) or restricting out‐of‐trial use but permitting coverage within trials such as Medicare's Coverage with Study Participation (CSP).MethodsWe compared the financial impact to manufacturers and insurers of three regulatory alternatives: (1) limited regulation (current approach), (2) CSP, and (3) restrictive regulation (like the current drug approval process). Using data for patent foramen ovale closure devices, we modeled key parameters including recruitment time, probability of device efficacy, market adoption, and device cost/price to calculate profits to manufacturers, costs to insurers, and overall societal impact on health.ResultsFor manufacturers, profits were greatest under CSP—driven by faster market adoption of effective devices—followed by restrictive regulation. Societal health benefit in total quality‐adjusted life years was greatest under CSP. Insurers’ expenditures for ineffective devices were greatest with limited regulation. Findings were robust over a reasonable range of probabilities of trial success.ConclusionsRegulation restricting out‐of‐trial device use and extending limited insurance coverage to clinical trial participants may balance manufacturer and societal interests.
Evaluation of thyroid dysfunction and autoimmunity in gestational diabetes mellitus and its relationship with postpartum thyroiditis
Authors: Maleki N. , Tavosi Z. 
Published: September 2014

AbstractAimsTo evaluate thyroid dysfunction and autoimmunity in women with gestational diabetes and to investigate the frequency of postpartum thyroiditis in women with gestational diabetes.Materials and methodsA total of 350 women with gestational diabetes and 350 healthy pregnant women were enrolled in the study. We studied the thyroid hormone profiles of the women in each group during pregnancy (at 24–28 weeks' gestation) and after delivery (at 6 weeks, 3, 6 and 9 months, and 1 year postpartum).ResultsA total of 342 women with gestational diabetes and 313 healthy pregnant women completed the follow‐up during pregnancy and 1 year after delivery. Of the women with gestational diabetes, 16.6% had thyroid dysfunction, while of the healthy pregnant women, 6.1% had thyroid dysfunction. The prevalence of postpartum thyroiditis was higher in the women with a history of gestational diabetes (19.6%) than in the healthy pregnant women (10.2%), and this difference was statistically significant.ConclusionAccording to the results of the present study, the prevalence of postpartum thyroiditis was higher in women with a history of gestational diabetes than in healthy women. We recommend that all women with gestational diabetes and women who have previous thyroid dysfunction should be screened for thyroid hormonal abnormalities during pregnancy and for 1 year after pregnancy.
In vivo corneal confocal microscopy as a novel non‐invasive tool to investigate cardiac autonomic neuropathy in Type 1 diabetes
Authors: Maddaloni E. , Sabatino F. , Del Toro R. , Crugliano S. , Grande S. , Lauria Pantano A. , Maurizi A. R. , Palermo A. , Bonini S. , Pozzilli P. , Manfrini S. 
Published: September 2014

AbstractAimsTo investigate whether small nerve fibre degeneration detected using corneal confocal microscopy is associated with cardiac autonomic neuropathy in people with Type 1 diabetes.MethodsThirty‐six people with Type 1 diabetes and 20 age‐ and sex‐matched healthy control subjects were enrolled. Tests to determine heart rate response to deep‐breathing (expiratory‐to‐inspiratory ratio), heart rate response to lying‐to‐stand test (30:15 ratio) and blood pressure response to standing were performed to detect cardiac autonomic neuropathy. Corneal confocal microscopy was performed to assess: corneal nerve density and corneal nerve beadings; branching pattern; and nerve fibre tortuosity.ResultsCompared with control participants, participants with Type 1 diabetes had fewer (mean ± sd 45.4 ± 20.2 vs 92.0 ± 22.7 fibres/mm2; P < 0.001) and more tortuous corneal nerve fibres (20 participants with Type 1 diabetes vs four control participants had nerve tortuosity grade 2/3; P = 0.022) and fewer beadings (mean ± sd 15.1 ± 3.5 vs 20.6 ± 5.0; P < 0.001). Of the participants with Type 1 diabetes, 11 met the criteria for the diagnosis of cardiac autonomic neuropathy. Corneal nerve density was significantly lower in participants with cardiac autonomic neuropathy than in those without (mean ± sd 32.8 ± 16.4 vs 51.7 ± 18.9 fibres/mm2; P = 0.008). This difference remained significant after adjustment for age (P = 0.02), gender (P = 0.04), disease duration (P = 0.005), insulin requirement (P = 0.02) and neuropathy disability score (P = 0.04).ConclusionThis study suggests that corneal confocal microscopy could represent a new and non‐invasive tool to investigate cardiac autonomic neuropathy in people with Type 1 diabetes. Larger studies are required to define the role of corneal confocal microscopy in the assessment of cardiac autonomic neuropathy.
Excessive fluoride reduces Foxo1 expression in dental epithelial cells of the rat incisor
Authors: Gao Jianghong , Ruan Jianping , Gao Liping 
Published: September 2014

Enamel fluorosis is characterized by hypomineralization, and forkhead box O1 (Foxo1) is essential for mouse enamel biomineralization. This study investigated the effect of fluoride on Foxo1 expression and its implications for enamel fluorosis. Mandibular incisors were extracted from Sprague Dawley rats treated for 3 months with water containing 0, 50, or 100 p.p.m. F−. Immunohistochemistry was used to localize and quantify FOXO1 expression in dental epithelial layer cells of the incisors. The effect of fluoride on expression of Foxo1, kallikrein‐4 (Klk4), and amelotin (Amtn) mRNAs was analyzed by real‐time RT‐PCR, and western blotting was used to measure total and nuclear FOXO1 protein levels in mature dental epithelial cells. The results revealed that nuclear FOXO1 was mainly localized in the transition and the mature ameloblasts and exhibited weaker expression in the rats exposed to fluoride. In addition to the reduced levels of Foxo1, Klk4, and AmtnmRNAs, the protein levels of total and nuclearFOXO1 were decreased in the mature dental epithelial cells exposed to fluoride. Thus, excessive fluoride may have an effect on the expression levels of Foxo1 in dental epithelial cells and thereby affect hypomineralization of the enamel during fluorosis.
Cellular damage suffered by equine embryos after exposure to cryoprotectants or cryopreservation by slow‐freezing or vitrification
Authors: Hendriks W. K. , Roelen B. A. J. , Colenbrander B. , Stout T. A. E. 
Published: September 2014

SummaryReasons for performing studyEquine embryos are cryopreserved by slow‐freezing or vitrification. While small embryos (<300 μm) survive cryopreservation reasonably well, larger embryos do not. It is not clear if slow‐freezing or vitrification is less damaging to horse embryos.ObjectivesTo compare the type and extent of cellular damage suffered by small and large embryos during cryopreservation by slow‐freezing vs. vitrification.Study designSixty‐three Day 6.5–7 embryos were subdivided by size and assigned to one of 5 treatments: control, exposure to slow‐freezing or vitrification cryoprotectants (CPs), and cryopreservation by either technique.MethodsAfter thawing/CPremoval, embryos were stained with fluorescent stains for various parameters of cellular integrity, and assessed by multiphoton microscopy.ResultsExposing large embryos to vitrificationCPsresulted in more dead cells (6.8 ± 1.3%: 95% confidence interval [CI], 3.1–10.4%) than exposure to slow‐freezing media (0.3 ± 0.1%; 95%CI0.0–0.6%: P = 0.001). Cryopreservation by either technique induced cell death and cytoskeleton disruption. Vitrification of small embryos resulted in a higher proportion of cells with fragmented or condensed (apoptotic) nuclei (P=0.002) than slow‐freezing (6.7 ± 1.5%, 95%CI3.0–10.4% vs. 5.0 ± 2.1%, 95%CI4.0–14.0%). Slow‐freezing resulted in a higher incidence of disintegrated embryos (P=0.01) than vitrification. Mitochondrial activity was low in control embryos, and was not differentially affected by cryopreservation technique, whereas vitrification changed mitochondrial distribution from a homogenous crystalline pattern in control embryos to a heterogeneous granulated distribution in vitrified embryos (P=0.05).ConclusionsCryopreservation caused more cellular damage to large embryos than smaller ones. While vitrification is more practical, it is not advisable for large embryos due to a higher incidence of dead cells. The choice is less obvious for small embryos, as vitrification led to occasionally very high percentages of dead or damaged cells, but a lower incidence of embryo disintegration. Modifications that reduce the level of cellular damage induced by vitrification are required before it can be considered the method of choice for cryopreserving equine embryos.
A MODFLOW Infiltration Device Package for Simulating Storm Water Infiltration
Authors: Jeppesen Jan , Christensen Steen 
Published: September 2014

AbstractThis article describes a MODFLOW Infiltration Device (INFD) Package that can simulate infiltration devices and their two‐way interaction with groundwater. The INFD Package relies on a water balance including inflow of storm water, leakage‐like seepage through the device faces, overflow, and change in storage. The water balance for the device can be simulated in multiple INFD time steps within a single MODFLOW time step, and infiltration from the device can be routed through the unsaturated zone to the groundwater table. A benchmark test shows that the INFD Package's analytical solution for stage computes exact results for transient behavior. To achieve similar accuracy by the numerical solution of the MODFLOW Surface‐Water Routing (SWR1) Process requires many small time steps. Furthermore, the INFD Package includes an improved representation of flow through the INFD sides that results in lower infiltration rates than simulated by SWR1. The INFD Package is also demonstrated in a transient simulation of a hypothetical catchment where two devices interact differently with groundwater. This simulation demonstrates that device and groundwater interaction depends on the thickness of the unsaturated zone because a shallow groundwater table (a likely result from storm water infiltration itself) may occupy retention volume, whereas a thick unsaturated zone may cause a phase shift and a change of amplitude in groundwater table response to a change of infiltration. We thus find that the INFD Package accommodates the simulation of infiltration devices and groundwater in an integrated manner on small as well as large spatial and temporal scales.
Acute Diffusion-Weighted Imaging Lesion Patterns Predict Progressive Small Subcortical Infarct in the Perforator Territory of the Middle Cerebral Artery
Authors: Duan Zuowei , Sun Wen , Liu Wenhua , Xiao Lulu , Huang Zhixin , Cao Liping , Li Hua , Xiong Yunyun , Liu Dezhi , Xu Gelin , Liu Xinfeng 
Published: September 2014

Background Single small subcortical infarct (SSSI) is generally considered to have a fair outcome. However, early neurological deterioration (END), a relatively unfavorable clinical course occurring during the acute phase of infarction, is not uncommon. Aims The aim of this study was to investigate the relationship between lesion patterns detected by diffusion-weighted imaging (DWI) and the presence of END in patients with acute SSSI in the perforator territory of the middle cerebral artery (MCA). Methods Three hundred twelve patients with acute SSSI in the perforator territory of MCA were prospectively recruited from Jinling Hospital between January 2010 and May 2013. Acute DWI lesion patterns were classified as proximal SSSI (pSSSI) or distal SSSI (dSSSI) patterns, according to the relationship between lesion location and the parent artery. Neurological deficits were evaluated using the National Institutes of Health Stroke Scale (NIHSS) at admission and continued over the following 72 h 1–3 times a day. END was defined as an increase in NIHSS score ⩾2 points during the first 72 h after admission. Results Of the total 312 patients, the pSSSI pattern was found in 139 (44·55%) patients and the dSSSI pattern in 173 (55·45%) patients. Statistical analysis suggested that the indicators for small-artery disease (hypertension and leukoaraiosis) and atherosclerosis (diabetes mellitus and cerebral atherosclerosis) significantly differed according to lesion patterns ( P < 0·05). During hospitalization, 88 (28·21%) patients experienced END. Univariate analysis revealed that female sex ( P = 0·004), pSSSI pattern ( P < 0·001), initial NIHSS ( P = 0·001), lesion diameter ( P = 0·005), ipsilateral large-artery stenosis ( P = 0·008), and concomitant intracranial atherosclerotic stenosis ( P = 0·021) were significantly associated with END. After adjusting for confounding factors, pSSSI pattern was an independent predictor of END (OR 1·871, 95% CI 1·095–3·198, P = 0·022). In the further subgroup analysis of patients with different etiologies, pSSSI pattern was found to be independently associated with END in patients with large-artery atherosclerosis (OR 3·593, 95% CI 1·268–11·057, P = 0·026) and in patients with small-artery disease (OR 2·523, 95% CI 1·121–5·676, P = 0·025), but not in patients with cardioembolism (OR 0·854, 95% CI 0·147–4·953, P = 0·861). Conclusions pSSSI pattern was closely related to END in acute SSSI caused by large-artery atherosclerosis and small-artery disease in the perforator territory of the MCA.