Profiling Additional Effects of Aptamer Fluorophore Modification on Microscale Thermophoresis Characterization of Aptamer-Target Binding.
Aptamers are promising affinity ligands with considerable applications, such as biosensors, disease diagnosis, therapy, etc. Characterization of aptamer-target binding is important in aptamer selection and aptamer applications. Microscale thermophoresis (MST) is an emerging optical technique for molecular interactions, which monitors fluorescence responses of fluorescent molecules in a microscopic temperature gradient. Harnessing merits in trace sample consumption, high speed, free separation, free immobilization, and ratiometric analysis, MST draws intense wide attention. MST is often applied for aptamer-target binding studies using fluorescently labeled aptamers. However, the MST signal is strongly dependent on fluorophore modifications at aptamers, which brings additional challenges and effects for MST analyzing aptamer affinity. Here, we systematically explored effects of fluorophore modifications (e.g., fluorophore types, fluorophore positions, etc.) of aptamer probes on MST characterizing aptamer-target interactions and identified gaps of MST analysis in aptamer affinity determination, taking aptamers against cadmium ions and aflatoxin B1 as two representatives. The same aptamers with different fluorophore modifications showed distinct MST signals in response magnitudes and signs as well as determined affinities, and some of them failed to respond to target binding and gave false affinity information in MST. A competitive MST method can be used to extract the affinity of unmodified aptamers, excluding effects of fluorophore modification. This work highlights that appropriate fluorophore modification is crucial in MST analysis of aptamer affinity, and caution is needed in MST applications, providing a basis for rational design of the MST method for the reliable molecular interaction study.


Long-term operation of microbial electrosynthesis cell reducing CO2 to multi-carbon chemicals with a mixed culture avoiding methanogenesis.
In microbial electrosynthesis (MES), CO2 can be reduced preferably to multi-carbon chemicals by a biocathode-based process which uses electrochemically active bacteria as catalysts. A mixed anaerobic consortium from biological origin typically produces methane from CO2 reduction which circumvents production of multi-carbon compounds. This study aimed to develop a stable and robust CO2 reducing biocathode from a mixed culture inoculum avoiding the methane generation. An effective approach was demonstrated based on (i) an enrichment procedure involving inoculum pre-treatment and several culture transfers in H2:CO2 media, (ii) a transfer from heterotrophic to autotrophic growth and (iii) a sequential batch operation. Biomass growth and gradual acclimation to CO2 electro-reduction accomplished a maximum acetate production rate of 400mgLcatholyte-1d-1 at -1V (vs. Ag/AgCl). Methane was never detected in more than 300days of operation. Accumulation of acetate up to 7-10gL-1 was repeatedly attained by supplying (80:20) CO2:N2 mixture at -0.9 to -1V (vs. Ag/AgCl). In addition, ethanol and butyrate were also produced from CO2 reduction. Thus, a robust CO2 reducing biocathode can be developed from a mixed culture avoiding methane generation by adopting the specific culture enrichment and operation procedures without the direct addition of chemical inhibitor.


Modulators of crescentic glomerulonephritis.
Glomerular crescent formation is a prominent feature of aggressive forms of glomerulonephritis and is associated with a poor prognosis. An understanding of the mechanisms involved in crescent formation is crucial for the development of new therapies for this disease. This article reviews current ideas on the pathogenesis of glomerular crescent formation and describes methods for modulation of this process. Emphasis is given to the role of the proinflammatory cytokines interleukin-1 and tumor necrosis factor-alpha in crescent development and its modulation by cytokine blockade.


Differences in the phenotype, cytokine gene expression profiles, and in vivo alloreactivity of T cells mobilized with plerixafor compared with G-CSF.
Plerixafor (Mozobil) is a CXCR4 antagonist that rapidly mobilizes CD34(+) cells into circulation. Recently, plerixafor has been used as a single agent to mobilize peripheral blood stem cells for allogeneic hematopoietic cell transplantation. Although G-CSF mobilization is known to alter the phenotype and cytokine polarization of transplanted T cells, the effects of plerixafor mobilization on T cells have not been well characterized. In this study, we show that alterations in the T cell phenotype and cytokine gene expression profiles characteristic of G-CSF mobilization do not occur after mobilization with plerixafor. Compared with nonmobilized T cells, plerixafor-mobilized T cells had similar phenotype, mixed lymphocyte reactivity, and Foxp3 gene expression levels in CD4(+) T cells, and did not undergo a change in expression levels of 84 genes associated with Th1/Th2/Th3 pathways. In contrast with plerixafor, G-CSF mobilization decreased CD62L expression on both CD4 and CD8(+) T cells and altered expression levels of 16 cytokine-associated genes in CD3(+) T cells. To assess the clinical relevance of these findings, we explored a murine model of graft-versus-host disease in which transplant recipients received plerixafor or G-CSF mobilized allograft from MHC-matched, minor histocompatibility-mismatched donors; recipients of plerixafor mobilized peripheral blood stem cells had a significantly higher incidence of skin graft-versus-host disease compared with mice receiving G-CSF mobilized transplants (100 versus 50%, respectively, p = 0.02). These preclinical data show plerixafor, in contrast with G-CSF, does not alter the phenotype and cytokine polarization of T cells, which raises the possibility that T cell-mediated immune sequelae of allogeneic transplantation in humans may differ when donor allografts are mobilized with plerixafor compared with G-CSF.


Identification of structural domains in inter-alpha-trypsin involved in calcium oxalate crystallization.
The urinary glycoprotein that inhibits calcium oxalate (CaOx) crystallization in vitro shows a structural similarity to urinary trypsin inhibitor (UTI; recently termed bikunin), the light chain of inter-alpha-trypsin inhibitor (I alpha I). The functional domains of I alpha I involved in its inhibitory activity of CaOx crystallization have been investigated using isolated intact domains of I alpha I produced from controlled proteolytic digests of the protein. The fragments investigated include the heavy chains of I alpha I, UTI, chondroitinase AC-treated UTI, and the carboxyl-terminal domain of UTI (termed HI-8). The effects of I alpha I and its fragments on the inhibitory activity of CaOx crystallization were evaluated in vitro using CaOx crystal aggregation and growth assays, and seeded crystal generation assay as well as using crystal matrix protein generation assay. UTI, but not the heavy chains of I alpha I, had a discernible effect on CaOx crystallization inhibitory activity. Less requirement of the carbohydrate moiety of UTI is implicated by the observation that chondroitinase AC-treated UTI fragment was also found to inhibit CaOx crystallization with almost the same activity as UTI. HI-8 also efficiently inhibited CaOx crystallization, while I alpha I showed a weak inhibitory activity. The results are almost consistent with a seed crystal generation assay and a crystal adsorption inhibition assay, in which I alpha I or its derivatives inhibits prothrombin fragment 1 (F1) adsorption to CaOx crystals. In conclusion, these results suggest that the part of the I alpha I protein responsible for inhibition of CaOx crystallization is the carboxyl-terminal domain of UTI.


Structural aspects of RimP binding on small ribosomal subunit from Staphylococcus aureus.
Ribosome biogenesis is an energy-intense multistep process where even minimal defects can cause severe phenotypes up to cell death. Ribosome assembly is facilitated by biogenesis factors such as ribosome assembly factors. These proteins facilitate the interaction of ribosomal proteins with rRNA and correct rRNA folding. One of these maturation factors is RimP which is required for efficient 16S rRNA processing and 30S ribosomal subunit assembly. Here, we describe the binding mode of Staphylococcus aureus RimP to the small ribosomal subunit and present a 4.2 Å resolution cryo-EM reconstruction of the 30S-RimP complex. Together with the solution structure of RimP solved by NMR spectroscopy and RimP-uS12 complex analysis by EPR, DEER, and SAXS approaches, we show the specificity of RimP binding to the 30S subunit from S. aureus. We believe the results presented in this work will contribute to the understanding of the RimP role in the ribosome assembly mechanism.


Modulation of Kv4-encoded K(+) currents in the mammalian myocardium by neuronal calcium sensor-1.
Voltage-gated K(+) channels are multimeric proteins, consisting of four pore-forming alpha-subunits alone or in association with accessory subunits. Recently, for example, it was shown that the accessory Kv channel interacting proteins form complexes with Kv4 alpha-subunits and modulate Kv4 channel activity. The experiments reported here demonstrate that the neuronal calcium sensor protein-1 (NCS-1), another member of the recoverin-neuronal calcium sensor superfamily, is expressed in adult mouse ventricles and that NCS-1 co-immunoprecipitates with Kv4.3 from (adult mouse) ventricular extracts. In addition, co-expression studies in HEK-293 cells reveal that NCS-1 increases membrane expression of Kv4 alpha-subunits and functional Kv4-encoded K(+) current densities. Co-expression of NCS-1 also decreases the rate of inactivation of Kv4 alpha-subunit-encoded K(+) currents. In contrast to the pronounced effects of Kv channel interacting proteins on Kv4 channel gating, however, NCS-1 co-expression does not measurably affect the voltage dependence of steady-state inactivation or the rate of recovery from inactivation of Kv4-encoded K(+) currents. Taken together, these results suggest that NCS-1 is an accessory subunit of Kv4-encoded I(to,f) channels that functions to regulate I(to,f) density in the mammalian myocardium.


Implementing CUREs with Cookbook-Style Laboratory Exercises in In-Person, Online, and Hybrid Formats.
Course-based undergraduate research experiences (CUREs) provide students with ample opportunity to engage in the scientific process and are increasingly replacing more traditional cookbook-style laboratory exercises in the undergraduate biology curriculum. The COVID-19 pandemic has highlighted the difficulty of implementing these projects during times of crisis. The quick transition to online learning during the pandemic afforded us a unique opportunity to develop an alternative version of our CURE for the online environment and to compare the efficacy of our CURE on learning gains in online versus in-person learning situations. Compared with a previous semester, our data suggest no significant differences in learning gains between students who participated in a hands-on CURE and an online CURE. Taken together the data suggests an avenue for CURE implementation even while teaching in online or hybrid formats. We discuss strategies that made the CURE successful in the online format and offer suggestions for how to adapt such activities for online or hybrid courses.


Testing and linkage to care outcomes for a clinician-initiated rapid HIV testing program in an urban emergency department.
The urban emergency department is an important site for the detection of HIV infection. Current research has focused on strategies to increase HIV testing in the emergency department. As more emergency department HIV cases are identified, there need to be well-defined systems for linkage to care. We conducted a retrospective study of rapid HIV testing in an urban public emergency department and level I trauma center from June 1, 2008, to March 31, 2010. The objectives of this study were to evaluate the increase in the number of tests and new HIV diagnoses resulting from the addition of targeted testing to clinician-initiated diagnostic testing, describe the demographic and clinical characteristics of patients with newly diagnosed HIV infection, and assess the effectiveness of an HIV clinic based linkage to care team. Of 96,711 emergency department visits, there were 5340 (5.5%) rapid HIV tests performed, representing 4827 (91.3%) unique testers, of whom 62.4% were male and 60.8% were from racial/ethnic minority groups. After the change in testing strategy, the median number of tests per month increased from 114 to 273 (p=0.004), and the median number of new diagnoses per month increased from 1.5 to 4 (p=0.01). From all tests conducted, there were 65 new diagnoses of HIV infection (1.2%, 95% confidence interval [CI] 0.9%, 1.5%). The linkage team connected over 90% of newly diagnosed and out-of-care HIV-infected patients to care. In summary, the addition of targeted testing to diagnostic testing increased new HIV case identification, and an HIV clinic-based team was effective at linkage to care.


Formulation and In-vitro Evaluation of pH-Sensitive Oil Entrapped Polymeric Blend Amoxicillin Beads for the Eradication of Helicobacter pylori.
Oral pH sensitive drug delivery systems are of utmost importance as these systems deliver the drug at specific part of the gastrointestine (GI) as per the pH of GI, resulting in improved patient therapeutic efficacy and compliance. The pH range of fluids in various segments of the GI tract may provide environmental stimuli for drug release. The aim of this study was to design buoyant beads containing amoxicillin (Am) and to evaluate its potential for the eradication of Helicobacter pylori (H. pylori). The gel bead of gellan, wherein the oil was entrapped, was blended with hydroxypropyl methyl cellulose or Carbopol 934. Buoyant beads of gellan were prepared through ionotropic gellation technique to achieve the controlled and pH-sensitive drug release in stomach. The effects of processing variables such as particle size, buoyancy, percent encapsulation efficiency and in-vitro antimicrobial activity were evaluated. The scanning electron micrograph indicated that prepared beads were spherical in shape and all the beads showed satisfactory floating efficiency in the phthalate buffer solution. The diameter of the gel beads was increased through raising the gellan gum and calcium carbonate concentration. The formulation exhibited sustained release profile and was best fitted in the Peppas model with n < 0.45. Subsequent coating of microbeads exhibited zero-order sustained pattern of the drug release up to 8 h. In-vitro growth inhibition study showed complete eradication of the isolated H. pylori strain .These results provide evidence that the optimized formulation bearing antibiotics like amoxicillin should be useful in H. pylori treatment.


Perineal colostomy prolapse: a novel application of mesh sacral pexy.
Full thickness colonic prolapse following pseudocontinent perineal colostomy has not been previously reported. Possible contributing factors include a large skin aperture at the site of the perineal stoma, the absence of anal sphincters and mesorectal attachments and the presence of a perineal hernia. A novel application of sacral pexy combined with perineal hernia repair using two prosthetic meshes is described.


The Bigger Fish: A Comparison of Meta-Learning QSAR Models on Low-Resourced Aquatic Toxicity Regression Tasks.
Toxicological information as needed for risk assessments of chemical compounds is often sparse. Unfortunately, gathering new toxicological information experimentally often involves animal testing. Simulated alternatives, e.g., quantitative structure-activity relationship (QSAR) models, are preferred to infer the toxicity of new compounds. Aquatic toxicity data collections consist of many related tasks─each predicting the toxicity of new compounds on a given species. Since many of these tasks are inherently low-resource, i.e., involve few associated compounds, this is challenging. Meta-learning is a subfield of artificial intelligence that can lead to more accurate models by enabling the utilization of information across tasks. In our work, we benchmark various state-of-the-art meta-learning techniques for building QSAR models, focusing on knowledge sharing between species. Specifically, we employ and compare transformational machine learning, model-agnostic meta-learning, fine-tuning, and multi-task models. Our experiments show that established knowledge-sharing techniques outperform single-task approaches. We recommend the use of multi-task random forest models for aquatic toxicity modeling, which matched or exceeded the performance of other approaches and robustly produced good results in the low-resource settings we studied. This model functions on a species level, predicting toxicity for multiple species across various phyla, with flexible exposure duration and on a large chemical applicability domain.


[History of the study, epidemiology of the disease and characterization of Borrelia burgdorferi infection].
From the beginning of the twentieth century interest in tick-borne borreliosis continues to be an important diagnostic and therapeutic problem. In 1977, in Lyme and Old Lyme, Cunnecticut, in the United States, arthritis and skin lesions were described in 12 children, which were associated with tick bites. The name of the place was called a syndrome of Lyme disease. Dr Willy Burgdorfer observed microaerophilic Gram-negative bacteria (Spirochetes) with a diameter of 0.3-0.5 µm and a length of about 20-30 µm, which were isolated from the tick colon. Lyme disease (borreliosis) is the most common tick-borne disease in the world. In Poland, most cases of Lyme disease occur between May and November, and more than 80% of cases are found in June and July during the nymph ticks feeding. The northern part of the country is considered endemic. The reservoir of bacteria is about 300 different species of mammals. These are mainly rodents of the norovirus and mouse family, and live deer, deer and wolves, as well as some species of birds. Ticks are infected with Borelia burgdorferi through the blood of infected animals. However, adult ticks are infested with Lyme spirochetes, depending on the region of the country. It infects 5-20% nymph and 15-40% adult individuals. The sequence of the Lyme disease spirochetes reveals a number of interesting traits. Classical virulence factors have not been isolated and therefore it is suspected that the vertebrate does not contain it in relation to mammals. However, a number of lipoproteins have been defined and some of them may be factors contributing to the innate immune system.


Out-of-hours discharge from intensive care, in-hospital mortality and intensive care readmission rates: a systematic review and meta-analysis.
Discharge from an intensive care unit (ICU) out of hours is common. We undertook a systematic review and meta-analysis to explore the association between time of discharge and mortality/ICU readmission. We searched Medline, Embase, Web of Knowledge, CINAHL, the Cochrane Library and OpenGrey to June 2017. We included studies reporting in-hospital mortality and/or ICU readmission rates by ICU discharge "out-of-hours" and "in-hours". Inclusion was limited to patients aged ≥ 16 years discharged alive from a non-specialist ICU to a lower level of hospital care. Studies restricted to specific diseases were excluded. We assessed study quality using the Newcastle Ottowa Scale. We extracted published data, summarising using a random-effects meta-analysis. Our searches identified 1961 studies. We included unadjusted data from 1,191,178 patients from 18 cohort studies (presenting data from 1994 to 2014). "Out of hours" had multiple definitions, beginning between 16:00 and 22:00 and ending between 05:59 and 09:00. Patients discharged out of hours had higher in-hospital mortality [relative risk (95% CI) 1.39 (1.24, 1.57) p < 0.0001] and readmission rates [1·30 (1.19, 1.42), p < 0.001] than patients discharged in hours. Heterogeneity was high (I2 90.1% for mortality and 90.2% for readmission), resulting from differences in effect size rather than the presence of an effect. Out-of-hours discharge from an ICU is strongly associated with both in-hospital death and ICU readmission. These effects persisted across all definitions of "out of hours" and across healthcare systems in different geographical locations. Whether these increases in mortality and readmission result from patient differences, differences in care, or a combination remains unclear.


Evaluation of the pharmacokinetic effects of various linking group using the 111In-DOTA-X-BBN(7-14)NH2 structural paradigm in a prostate cancer model.
The high incidence of BB2 receptor (BB2r) expression in various cancers has prompted investigators to pursue the development of BB2r-targeted agents for diagnostic imaging, chemotherapy, and radiotherapy. Development of BB2r-targeted agents, based on the bombesin (BBN) peptide, has largely involved the use of the bifunctional chelate approach in which the linking group serves several key roles including pharmacokinetic modification. Understanding the in vivo properties of the various pharmacokinetic modifying linking groups is crucial for developing BB2r-targeted agents with improved targeting and clearance characteristics. The goal of this study was to systematically evaluate the pharmacokinetic profile of aliphatic hydrocarbon, aromatic, and poly(ethylene glycol) (ether) functional groups in order to obtain a better understanding of the in vivo properties of these pharmacokinetic modifiers. Specifically, we synthesized six radioconjugates with the structure 111In-DOTA- X-BBN(7-14)NH2, where X = 8-aminooctanoic acid (8-AOC), 5-amino-3-oxapentyl-succinamic acid (5-ADS), 8-amino-3,6-dioxaoctyl-succinamic acid (8-AOS), p-aminobenzoic acid (AMBA), Gly-AMBA, and Gly- p-aminomethylbenzoic acid (Gly-AM2BA). All of the (nat)In-conjugates demonstrated nanomolar binding affinities to the BB2r. In CF-1 mice, the BB2r uptake in the pancreas of radioconjugates containing aromatic linking groups was found to be significantly higher at 1 h postinjection than the radioconjugates with ether linker moieties. For PC-3 tumor-bearing SCID mice, the tumor uptake was found to be 6.66 +/- 2.00, 6.21 +/- 1.57, 6.36 +/- 1.60, 4.46 +/- 0.81, and 7.76 +/- 1.19 %ID/g for the 8-AOC, 8-ADS, AMBA, Gly-AMBA, and Gly-AM2BA radioconjugates, respectively, at 15 min postinjection. By 24 h postinjection, the radioconjugates containing aromatic groups exhibited the highest percentage tumor retention with 11.4%, 19.8%, 26.6%, 25.8%, and 25.5% relative to the 15 min values remaining in the tumor tissue for the 8-AOC, 8-ADS, AMBA, Gly-AMBA, and Gly-AM2BA radioconjugates, respectively. Fused Micro-SPECT/CT imaging studies performed at 24 h postinjection revealed substantial accumulation of radioactivity in the tumor tissue for all radioconjugates. In both biodistribution and Micro-SPECT/CT imaging studies, the radioconjugates containing aromatic linking groups typically exhibited significantly higher G.I. tract retention than the hydrocarbon or ether linking moieties. In conclusion, our studies indicate that radioconjugates incorporating aromatic linking groups, of the type investigated, generally demonstrated enhanced retention in BB2r expressing tissues in comparison to either the hydrocarbon or ether linking moieties. Furthermore, this investigation clearly demonstrates the significance of the linking group upon not only the in vivo clearance of the radiopharmaceutical, but also on the in vivo uptake and retention of the BB2r-targeted agent in tumor tissue. Future designs of BB2r-targeted agents should include a careful consideration of the effect linking group functionality has upon tumor targeting and retention.


Impact of tumour bed boost integration on acute and late toxicity in patients with breast cancer: A systematic review.
The purpose of this systematic review was to summarise the evidence from studies investigating the integration of tumour bed boosts into whole breast irradiation for patients with Stage 0-III breast cancer, with a focus on its impact on acute and late toxicities. A comprehensive systematic electronic search through the Ovid MEDLINE, EMBASE and PubMed databases from January 2000 to January 2015 was conducted. Studies were considered eligible if they investigated the efficacy of hypo- or normofractionated whole breast irradiation with the inclusion of a daily concurrent boost. The primary outcomes of interest were the degree of observed acute and late toxicity following radiotherapy treatment. Methodological quality assessment was performed on all included studies using either the Newcastle-Ottawa Scale or a previously published investigator-derived quality instrument. The search identified 35 articles, of which 17 satisfied our eligibility criteria. Thirteen and eleven studies reported on acute and late toxicities respectively. Grade 3 acute skin toxicity ranged from 1 to 7% whilst moderate to severe fibrosis and telangiectasia were both limited to 9%. Reported toxicity profiles were comparable to historical data at similar time-points. Studies investigating the delivery of concurrent boosts with whole breast radiotherapy courses report safe short to medium-term toxicity profiles and cosmesis rates. Whilst the quality of evidence and length of follow-up supporting these findings is low, sufficient evidence has been generated to consider concurrent boost techniques as an alternative to conventional sequential techniques.


Fifty Years of Research in ARDS. Gas Exchange in Acute Respiratory Distress Syndrome.
Acute respiratory distress syndrome (ARDS) is characterized by severe impairment of gas exchange. Hypoxemia is mainly due to intrapulmonary shunt, whereas increased alveolar dead space explains the alteration of CO2 clearance. Assessment of the severity of gas exchange impairment is a requisite for the characterization of the syndrome and the evaluation of its severity. Confounding factors linked to hemodynamic status can greatly influence the relationship between the severity of lung injury and the degree of hypoxemia and/or the effects of ventilator settings on gas exchange. Apart from situations of rescue treatment, targeting optimal gas exchange in ARDS has become less of a priority compared with prevention of injury. A complex question for clinicians is to understand when improvement in oxygenation and alveolar ventilation is related to a lower degree or risk of injury for the lungs. In this regard, a full understanding of gas exchange mechanism in ARDS is imperative for individualized symptomatic support of patients with ARDS.


Long-term Success With Diminished Opioid Prescribing After Implementation of Standardized Postoperative Opioid Prescribing Guidelines: An Interrupted Time Series Analysis.
To assess longitudinal prescribing patterns for patients undergoing urologic surgery in the nearly 2-year time frame before and after implementation of an evidence-based opioid prescribing guideline to accurately characterize the impact on postoperative departmental practices. Historical prescribing data for adults who underwent 21 urologic procedures at 3 academic institutions were used to derive a 4-tiered guideline for postoperative opioid prescribing. The guideline was implemented on January 16, 2018, and prescribing patterns including quantity of opioids prescribed (in oral morphine equivalents [OMEs]) and refill rates were compared for opioid-naïve patients undergoing urologic surgery before (January 1, 2016, through January 15, 2018; N=10,649) and after (January 16, 2018, through September 30, 2019; N=9422) guideline implementation. Univariate analysis was performed using Wilcoxon rank sum and χ2 tests. Cochran-Armitage trend tests and interrupted time series analysis were used to test for significance in the change in OMEs prescribed before vs after guideline implementation. The median quantity of opioids decreased from 150 OMEs (interquartile range, 0-225) before guideline implementation to 0 OMEs (interquartile range, 0-90) after guideline implementation (P<.001). Median OMEs decreased significantly in each tier and each of 21 individual procedures. Overall guideline adherence was 90.7% (n=8547). Despite this decrease in OMEs prescribed, post-guideline implementation patients obtained fewer refills than the pre-guideline implementation group (614 [6.5%] vs 999 [9.4%]; P<.001). In a multi-institutional follow-up prospective study of adult urologic surgery-specific evidence-based guidelines for postoperative prescribing, we demonstrate sustained reduction in OMEs prescribed secondary to guideline implementation and adherence by our providers.


Pregnancy outcome and Ebstein's anomaly.
Ebstein's anomaly is an uncommon congenital cardiac abnormality that may be associated with cyanosis and arrhythmias. For those female patients with the anomaly who survive to adult life there is little information available about pregnancy, maternal complications, and fetal outcome. This study was designed to address this issue so that these patients can receive appropriate advice and management. Forty two pregnancies in 12 women with Ebstein's anomaly were studied. The mothers' cardiac lesions were assessed on the basis of symptoms, the presence of cyanosis or arrhythmia, and by echocardiographic grading of severity. In the absence of important maternal cyanosis or arrhythmia, pregnancy was well tolerated. Neonatal outcome was good though there was an increased risk of prematurity and dysmaturity in the babies born to mothers with cyanosis. This study indicates that women with Ebstein's anomaly who reach child-bearing age can be advised that pregnancy is likely to be well tolerated with good fetal outcome. Maternal arrhythmia or cyanosis are indications for closer maternal and fetal observation.


Preparation of a magnetic molecularly imprinted polymer with pseudo template for rapid simultaneous determination of cyromazine and melamine in bio-matrix samples.
A magnetic molecularly imprinted polymer (M-MIP) for cyromazine and melamine was prepared by simple suspension polymerization using a pseudo template, 2-(4,6-diamino-1,3,5-triazin-2-ylamino)ethanethiol disulfide. The M-MIP was characterized by scanning electron microscopy, Fourier-transform infrared spectroscopy, and vibrating sample magnetometry. Molecular recognition properties and binding capability to cyromazine and melamine were evaluated by adsorption testing, which showed the M-MIP had better affinity and selectivity than the magnetic non-imprinted polymer (M-NIP) for cyromazine and melamine. A method based on molecularly imprinted solid-phase extraction assisted by magnetic separation was developed for extraction of cyromazine and melamine from bio-matrix samples. Various conditions, for example desorption conditions, amount of M-MIP, extraction time, and sample pH were optimized. High-performance liquid chromatography with UV detection was used to determine cyromazine and melamine after extraction. The proposed method was successfully applied to determination of cyromazine and melamine in egg and milk samples. Recovery of standard spiked cyromazine and melamine from these samples was between 71.86 and 80.57%, with intraday and interday relative standard deviation ranging from 3.45 to 6.39% and from 3.95 to 7.84%, respectively. The results indicate that the pseudo template M-MIP can be used for preconcentration, purification, and analysis of cyromazine and melamine in bio-matrix samples.


UV Radiation Activates Toll-Like Receptor 9 Expression in Primary Human Keratinocytes, an Event Inhibited by Human Papillomavirus 38 E6 and E7 Oncoproteins.
Several lines of evidence indicate that cutaneous human papillomavirus (HPV) types belonging to the beta genus of the HPV phylogenetic tree synergize with UV radiation in the development of skin cancer. Accordingly, the E6 and E7 oncoproteins from some beta HPV types are able to deregulate pathways related to immune response and cellular transformation. Toll-like receptor 9 (TLR9), in addition to playing a role in innate immunity, has been shown to be involved in the cellular stress response. Using primary human keratinocytes as experimental models, we have shown that UV irradiation (and other cellular stresses) activates TLR9 expression. This event is closely linked to p53 activation. Silencing the expression of p53 or deleting its encoding gene affected the activation of TLR9 expression after UV irradiation. Using various strategies, we have also shown that the transcription factors p53 and c-Jun are recruited onto a specific region of the TLR9 promoter after UV irradiation. Importantly, the E6 and E7 oncoproteins from beta HPV38, by inducing the accumulation of the p53 antagonist ΔNp73α, prevent the UV-mediated recruitment of these transcription factors onto the TLR9 promoter, with subsequent impairment of TLR9 gene expression. This study provides new insight into the mechanism that mediates TLR9 upregulation in response to cellular stresses. In addition, we show that HPV38 E6 and E7 are able to interfere with this mechanism, providing another explanation for the possible cooperation of beta HPV types with UV radiation in skin carcinogenesis.IMPORTANCE Beta HPV types have been suggested to act as cofactors in UV-induced skin carcinogenesis by altering several cellular mechanisms activated by UV radiation. We show that the expression of TLR9, a sensor of damage-associated molecular patterns produced during cellular stress, is activated by UV radiation in primary human keratinocytes (PHKs). Two transcription factors known to be activated by UV radiation, p53 and c-Jun, play key roles in UV-activated TLR9 expression. The E6 and E7 oncoproteins from beta HPV38 strongly inhibit UV-activated TLR9 expression by preventing the recruitment of p53 and c-Jun to the TLR9 promoter. Our findings provide additional support for the role that beta HPV types play in skin carcinogenesis by preventing activation of specific pathways upon exposure of PHKs to UV radiation.


Differentiation of adipose-derived stromal vascular fraction culture cells into chondrocytes using the method of cell sorting with a mesenchymal stem cell marker.
The incidence of arthritic diseases is rapidly increasing in most advanced countries. Articular cartilage, which is the most important tissue in the joint, consists of chondrocytes and abundant extracellular matrix, including aggrecan, and shows poor self-repair. We studied the potential of stem cells in mouse subcutaneous adipose tissue as a source of cells to regenerate cartilage tissue. Analysis of adipose-derived stromal vascular fraction culture cells (ADSVFs) using mesenchymal stem cell markers showed that CD90-positive cells accounted for 93.8%, CD105-positive cells for 68.5%, and p75 neurotrophin receptor (p75NTR, CD271)-positive cells for 36.1%. These results indicate that cells positive for mesenchymal stem cell markers are present in ADSVFs. The CD105-positive or -negative cells were isolated from ADSVFs by magnetic cell separation (MACS), and the efficiency of differentiation into chondrocytes was compared with using three methods of pellet method, gel-coating method, and gel-embedding sheet method. Using the CD105-positive cells and the gel-embedding sheet method, aggrecan mRNA was detected about three times higher than pellet and gel-coating methods. The above data suggest that ADSVFs could be differentiated into chondrocyte-like cells in the gel-embedding sheet method and could be useful in regenerative medicine to treat cartilage defects or cartilage degenerative disease. The use of cells sorted by mesenchymal stem cell markers from adipose tissue would gain position in the repair of cartilage tissue.


Crenotalea thermophila gen. nov., sp. nov., a member of the family Chitinophagaceae isolated from a hot spring.
A thermophilic aerobic bacterium designated strain STH-1-Y1(T) was isolated from sulfur-turf in a Japanese hot spring (Okuhodaka hot spring, Gifu Pref.). Colonies of strain STH-1-Y1(T) were yellow and low convex morphology with a slightly irregular fringe. Cells were slender long rods, 0.4-0.6 µm wide and 1.2-3.0 µm long. The isolate was an obligate aerophilic organism, and could not grow by fermentation or nitrate respiration. The isolate had a thermophilic trait, and could grow at 35-60 °C and pH 5.5-7.5; maximum growth occurred at 55 °C and pH 7.0 with a doubling time of 1.9 h. The Biolog and API tests suggested that strain STH-1-Y1(T) was able to use various sugars such as glucose, lactose, mannose, maltose, trehalose, cellobiose and sucrose, but could not use sugar alcohols other than glycerol, i.e. adonitol, arabitol, erythritol, inositol, mannitol, sorbitol and xylitol. Lactate and glutamate could be used, but other fatty acids, i.e. acetate, citrate, propionate and succinate could not. Gelatin, casein, starch and glycogen were hydrolysed, but neither chitin nor agar was degraded. Cells lacked flexirubin and showed oxidase and catalase activities. The major respiratory quinone was menaquinone-7 (MK-7), and major cellular fatty acids were iso-C15 : 0, iso-C17 : 0 3-OH, iso-C17 : 0 and anteiso-C15 : 0. No unsaturated fatty acids were detected. Phylogenetic analysis based on 16S rRNA gene sequences revealed that strain STH-1-Y1(T) was closely related to the family Chitinophagaceae within the phylum Bacteroidetes. However, the isolate was evenly distant from all members in this family with sequence similarities of 87-89 %. These significantly low sequence similarities strongly suggested that strain STH-1-Y1(T) represents a novel species in a new genus of the family Chitinophagaceae within the phylum Bacteroidetes. Based on phenotypic and phylogenetic characteristics, the name Crenotalea thermophila gen. nov., sp. nov. is proposed. The type strain of the type species is STH-1-Y1(T) ( = JCM 11541(T) = DSM 14807(T)).


Serinicoccus profundi sp. nov., an actinomycete isolated from deep-sea sediment, and emended description of the genus Serinicoccus.
A Gram-reaction-positive bacterial strain of the genus Serinicoccus, designated MCCC 1A05965(T), was isolated from a deep-sea (5368 m) sediment of the Indian Ocean. Comparison of 16S rRNA gene sequences revealed that the isolate shared 97.6 % sequence similarity with Serinicoccus marinus JC1078(T), the type strain of the only described species of the genus Serinicoccus. The DNA-DNA relatedness between these two strains was 46.2 % (standard deviation 1.86 %). The cell wall contained alanine, glycine, serine, l-ornithine and glutamic acid, which corresponds to the description of the genus Serinicoccus. The acyl type of the glycan chain of the peptidoglycan was glycolyl. Other characteristics of strain MCCC 1A05965(T) were consistent with those of the genus Serinicoccus. Cells were coccoid, moderately halophilic, oxidase-negative, catalase-positive and non-spore-forming. The major menaquinone was MK-8(H(4)). The predominant cellular fatty acids were iso-C(15 : 0) (34.7 %) and iso-C(16 : 0) (17.0 %). The polar lipids were phosphatidylglycerol, diphosphatidylglycerol, phosphatidylcholine, phosphatidylinositol and an unknown glycolipid. The DNA G+C content was 72 mol%. Strain MCCC 1A05965(T) (=0714S6-1(T) =DSM 21363(T) =CGMCC 4.5582(T)) is assigned as the type strain of a novel species, for which the name Serinicoccus profundi sp. nov. is proposed.


Protecting your dental license.
In this second installment in our series about licensing, we explore how actions taken by governmental agencies other than criminal courts or professional disciplinary agencies, can affect your dental license.


A shotgun lipidomics study of a putative lysophosphatidic acid acyl transferase (PlsC) in Sinorhizobium meliloti.
A shotgun lipidomics approach was used to study the knockout mutant of a putative lysophosphatidic acyl acid transferase (PlsC) in order to delineate the function of this enzyme in Sinorhizobium meliloti. In plsC knockout mutant lipids that contained 16:0 and 16:1 fatty acids and their biosynthetically related cyclopropane fatty acid (cis-9,10-methylene hexadecanoic acid) decreased up to 93%. Tandem mass spectrometry experiments in the presence of added Li(+) showed that the putative PlsC (SMc00714) functioned as a lysophosphatidic acid acyl transferase specific for the transfer of C16 fatty acids to the sn-2 position of lipids. The levels of lipids containing C18 fatty acids were unaffected in plsC mutant, suggesting the presence of one or more fatty acyl transferases in the genome of S. meliloti with selectivity towards C18 fatty acids. Two non-phosphorus containing lipid classes, sulfoquinovosyldiacylglycerol and 1,2-diacylglyceryl-trimethylhomoserine lipids, showed similar decreases in C16 fatty acid content as phospholipids in plsC knockout mutant; these non-phosphorus containing lipids share a common biosynthetic origin with phospholipids, most likely involving phosphatidic acid. Ornithine lipids containing C16 fatty acids also showed decreased levels in PlsC knockout mutant, suggesting that PlsC is also involved in their biosynthesis.


Occupational exposure to power frequency magnetic fields and risk of non-Hodgkin lymphoma.
To investigate the risk of non-Hodgkin lymphoma (NHL) using a job-exposure matrix (JEM) to assess exposure to occupational magnetic fields at the power frequencies of 50/60 Hz. The study population consisted of 694 cases of NHL, first diagnosed between 1 January 2000 and 31 August 2001, and 694 controls from two regions in Australia, matched by age, sex and region of residence. A detailed occupational history was given by each subject. Exposure to power frequency magnetic fields was estimated using a population-based JEM which was specifically developed in the United States to assess occupational magnetic field exposure. The cumulative exposure distribution was divided into quartiles and adjusted odds ratios were calculated using the lowest quartile as the referent group. For the total work history, the odds ratio (OR) for workers in the upper quartile of exposure was 1.48 (95% CI 1.02 to 2.16) compared to the referent (p value for trend was 0.006). When the exposure was lagged by 5 years the OR was 1.59 (95% CI 1.07 to 2.36) (p value for trend was 0.003). Adjusting for other occupational exposures did not significantly alter the results. These findings provide weak support for the hypothesis that occupational exposure to 50/60 Hz magnetic fields increases the risk of NHL.


Investigating Initial Disclosures and Reactions to Unexpected, Positive HPV Diagnosis.
Initial disclosures of health conditions are critical communication moments. Existing research focuses on disclosers; integrating confidants into studies of initial disclosures is needed. Guided by the disclosure decision-making model (DD-MM; Greene, 2009), this study examined what diagnosed persons and confidants may say when faced with unexpected test results and unexpected disclosures, respectively. Participants (N = 151) recorded an audio-visual message for another person, after imagining that they or the other person had just received unexpected, positive HPV test results. The qualitative analysis revealed four themes: (1) impression management and social distance, (2) invisible symptoms and advice regarding future disclosures, (3) expressing and acknowledging emotional reactions, and (4) misunderstandings and lacking knowledge about HPV. These findings suggested that DD-MM may be a relevant framework for understanding not only when disclosers share, but what disclosers and confidants say in early conversations about new diagnoses. While disclosers' and confidants' messages showed marked similarities, important differences appeared. For example, confidants focused on assuaging disclosers' fear about the consequences, whereas disclosers expressed distress related to their uncertainty about the prognosis of an HPV infection and how to prepare for next steps. The discussion highlighted implications for the DD-MM, HPV disclosures, and future interventions.


Aberrant upregulation of the glycolytic enzyme PFKFB3 in CLN7 neuronal ceroid lipofuscinosis.
CLN7 neuronal ceroid lipofuscinosis is an inherited lysosomal storage neurodegenerative disease highly prevalent in children. CLN7/MFSD8 gene encodes a lysosomal membrane glycoprotein, but the biochemical processes affected by CLN7-loss of function are unexplored thus preventing development of potential treatments. Here, we found, in the Cln7∆ex2 mouse model of CLN7 disease, that failure in autophagy causes accumulation of structurally and bioenergetically impaired neuronal mitochondria. In vivo genetic approach reveals elevated mitochondrial reactive oxygen species (mROS) in Cln7∆ex2 neurons that mediates glycolytic enzyme PFKFB3 activation and contributes to CLN7 pathogenesis. Mechanistically, mROS sustains a signaling cascade leading to protein stabilization of PFKFB3, normally unstable in healthy neurons. Administration of the highly selective PFKFB3 inhibitor AZ67 in Cln7∆ex2 mouse brain in vivo and in CLN7 patients-derived cells rectifies key disease hallmarks. Thus, aberrant upregulation of the glycolytic enzyme PFKFB3 in neurons may contribute to CLN7 pathogenesis and targeting PFKFB3 could alleviate this and other lysosomal storage diseases.


The Timing of School Transitions and Early Adolescent Problem Behavior.
This longitudinal study investigates whether rural adolescents who transition to a new school in sixth grade have higher levels of risky behavior than adolescents who transition in seventh grade. Our findings indicate that later school transitions had little effect on problem behavior between sixth and ninth grades. Cross-sectional analyses found a small number of temporary effects of transition timing on problem behavior: Spending an additional year in elementary school was associated with higher levels of deviant behavior in the Fall of Grade 6 and higher levels of antisocial peer associations in Grade 8. However, transition effects were not consistent across waves and latent growth curve models found no effects of transition timing on the trajectory of problem behavior. We discuss policy implications and compare our findings with other research on transition timing.


Blood lipid response to 20 weeks of supervised exercise in a large biracial population: the HERITAGE Family Study.
We studied the effects of 20 weeks of supervised cycle-ergometer exercise on plasma lipids in 675 healthy, sedentary, normolipidemic white and black men and women aged 17 to 65 years, participating in the HERITAGE Family Study. Fasting plasma lipids were assessed twice at baseline and 24 and 72 hours after the last exercise session and adjusted for plasma volume changes. No significant differences from the mean baseline levels were observed for total, low-density lipoprotein (LDL), and very-low-density lipoprotein (VLDL) cholesterol and apolipoprotein B (Apo B). A significant reduction (P < .01) from baseline levels in plasma total and VLDL triglycerides was observed only in the 24-hour posttraining specimens, reflecting a response to the last bout of exercise. High-density lipoprotein (HDL) cholesterol increased 3.6% for the combined group, primarily due to an increase in HDL2, with an associated increase in Apo A-1 (P < .001). No significant differences were noted in the HDL response by sex, race, or age. An inverse correlation (r = -.241) was observed between the increase in HDL cholesterol and change in body fat only in men, and the increase in HDL cholesterol was unrelated to the change in maximal oxygen uptake (VO2max).


Enhanced Bioaccumulation and Transfer of Monomethylmercury through Periphytic Biofilms in Benthic Food Webs of a River Affected by Run-of-River Dams.
Run-of-river (ROR) power plants impound limited terrestrial areas compared to traditional hydropower plants with large reservoirs and are assumed to have reduced impacts on mercury cycling. We conducted a study on periphyton and benthic communities from different habitats of the St. Maurice River (Québec, Canada) affected by two ROR power plants and their effect on the bioaccumulation and biomagnification of monomethylmercury (MMHg). Proportion of total mercury as MMHg reached maximum values about 2.9 times higher in flooded sites compared to unflooded sites. Impoundment by ROR would therefore provide favorable environments for the growth of periphyton, which can produce and accumulate MMHg. Periphyton MMHg concentrations significantly explained concentrations in some benthic macroinvertebrates, reflecting a local transfer. Through the analysis of δ13C and δ15N signatures, we found that flooding, creating scattered lenthic habitats, led to modifications in trophic structures by the introduction of new organic matter sources. The computed trophic magnification slopes did not show significant differences in the transfer efficiency of MMHg between sectors, while intercepts of flooded sectors were higher. Increases in MMHg concentrations in flooded areas are likely due to the impoundment, combined with watershed disturbances, and the creation of small habitats favorable to periphyton should be included in future predictive models.


Targeting the leukemic stem cell: the Holy Grail of leukemia therapy.
Since the discovery of leukemic stem cells (LSCs) over a decade ago, many of their critical biological properties have been elucidated, including their distinct replicative properties, cell surface phenotypes, their increased resistance to chemotherapeutic drugs and the involvement of growth-promoting chromosomal translocations. Of particular importance is their ability to transfer malignancy to non-obese diabetic-severe combined immunodeficient (NOD-SCID) mice. Furthermore, numerous studies demonstrate that acute myeloid leukemia arises from mutations at the level of stem cell, and chronic myeloid leukemia is also a stem cell disease. In this review, we will evaluate the main characteristics of LSCs elucidated in several well-documented leukemias. In addition, we will discuss points of therapeutic intervention. Promising therapeutic approaches include the targeting of key signal transduction pathways (for example, PI3K, Rac and Wnt) with small-molecule inhibitors and specific cell surface molecules (for example, CD33, CD44 and CD123), with effective cytotoxic antibodies. Also, statins, which are already widely therapeutically used for a variety of diseases, show potential in targeting LSCs. In addition, drugs that inhibit ATP-binding cassette transporter proteins are being extensively studied, as they are important in drug resistance-a frequent characteristic of LSCs. Although the specific targeting of LSCs is a relatively new field, it is a highly promising battleground that may reveal the Holy Grail of cancer therapy.


Structural determinants for binding of sorting nexin 17 (SNX17) to the cytoplasmic adaptor protein Krev interaction trapped 1 (KRIT1).
Sorting nexin 17 (SNX17) is a member of the family of cytoplasmic sorting nexin adaptor proteins that regulate endosomal trafficking of cell surface proteins. SNX17 localizes to early endosomes where it directly binds NPX(Y/F) motifs in the cytoplasmic tails of its target receptors to mediate their rates of endocytic internalization, recycling, and/or degradation. SNX17 has also been implicated in mediating cell signaling and can interact with cytoplasmic proteins. KRIT1 (Krev interaction trapped 1), a cytoplasmic adaptor protein associated with cerebral cavernous malformations, has previously been shown to interact with SNX17. Here, we demonstrate that SNX17 indeed binds directly to KRIT1 and map the binding to the second Asn-Pro-Xaa-Tyr/Phe (NPX(Y/F)) motif in KRIT1. We further characterize the interaction as being mediated by the FERM domain of SNX17. We present the co-crystal structure of SNX17-FERM with the KRIT1-NPXF2 peptide to 3.0 Å resolution and demonstrate that the interaction is highly similar in structure and binding affinity to that between SNX17 and P-selectin. We verify the molecular details of the interaction by site-directed mutagenesis and pulldown assay and thereby confirm that the major binding site for SNX17 is confined to the NPXF2 motif in KRIT1. Taken together, our results verify a direct interaction between SNX17 and KRIT1 and classify KRIT1 as a SNX17 binding partner.


Bioactive compounds in cranberries and their biological properties.
Cranberries are healthy fruit that contribute color, flavor, nutritional value, and functionality. They are one of only three fruits native to America. Over the past decade, public interest for the North American cranberry (Vaccinium macrocarpon) has been rising with reports of their potential health benefits linked to the numerous phytochemicals present in the fruit--the anthocyanins, the flavonols, the flavan-3-ols, the proanthocyanidins, and the phenolic acid derivatives. The presence of these phytochemicals appears to be responsible for the cranberry property of preventing many diseases and infections, including cardiovascular diseases, various cancers, and infections involving the urinary tract, dental health, and Helicobacter pylori-induced stomach ulcers and cancers. Recent years have seen important breakthroughs in our understanding of the mechanisms through which these compounds exert their beneficial biological effects, yet these remain to be scientifically substantiated. In this paper these characteristics, as well as the antioxidant, radical scavenging, antibacterial, antimutagen, and anticarcinogen properties of cranberry major bioactive compounds are explained.


Waveguiding mechanism in tube lattice fibers.
Waveguiding mechanism and modal characteristics of hollow core fibers consisting of a single or a regular arrangement of dielectric tubes are investigated. These fibers have been recently proposed as low loss, broadband THz waveguides. By starting from a description in terms of coupling between air and dielectric modes in a single tube waveguide, a simple and useful model is proposed and numerically validated. It is able to predict dispersion curves, high and low loss spectral regions, and the conditions to ensure the existence of low loss regions. In addition, it allows a better understanding of the role of the geometrical parameters and of the dielectric refractive index. The model is then applied to improve the tradeoff between low loss and effectively single mode propagation, showing that the best results are obtained with a heptagonal arrangement of the tubes.


Increased fecal primary bile acids in multiple myeloma with engraftment syndrome diarrhea after stem cell transplant.
Autologous stem cell transplant (ASCT) for multiple myeloma (MM) is associated with diarrhea during the peri-transplant period. We aimed to appraise mechanisms of peri-ASCT diarrhea in a prospective, longitudinal study of patients with MM. We compared by repeated measures (RM)-ANOVA daily bowel movements (BMs) and consistency [7-point Bristol Stool Form Scale (BSFS)], fecal calprotectin (intestinal inflammation), 13C-mannitol excretion in urine 0-2 h (small intestinal permeability), fasting serum C4 (bile acid synthesis) and total and primary bile acid in stool samples during baseline, peri-transplant period (Days 5-7 after stem cell infusion), and after hematological recovery post-ASCT. The 12 (5F, 7M) patients' median age was 61 y (IQR 54.8-63.3). All participants reported increased BMs (increase of 2 and 1 per day with and without engraftment syndrome, respectively). There were no significant increases in serum C4, total fecal bile acids, or intestinal permeability. Relative to patients without engraftment syndrome, four participants with engraftment syndrome had looser stool consistency (mean 2.6 points higher BSFS compared to without engraftment syndrome), increased primary fecal bile acids relative to baseline (>33 µmol/L vs. 6 µmol/L without engraftment syndrome), and increased fecal calprotectin compared to baseline (313 μg/mL vs. 35.6 μg/mL without engraftment syndrome; p = 0.06). Engraftment syndrome post-ASCT is associated with increased fecal primary bile acids.


Glutamatergic inputs contribute to phasic activity in vasopressin neurons.
Many neurons in the CNS display rhythmic patterns of activity to optimize excitation-secretion coupling. However, the mechanisms of rhythmogenesis are only partially understood. Magnocellular vasopressin (VP) neurons in the hypothalamus display a phasic activity that consists of alternative bursts of action potentials and silent periods. Previous observations from acute slices of adult hypothalamus suggested that VP cell rhythmicity depends on intrinsic membrane properties. However, such activity in vivo is nonregenerative. Here, we studied the mechanisms of VP neuron rhythmicity in organotypic slice cultures that, unlike acute slices, preserve functional synaptic connections. Comparative analysis of phasic firing of VP neurons in vivo, in acute slices, and in the cultures revealed that, in the latter, the activity was closely related to that observed in vivo. It was synaptically driven, essentially from glutamatergic inputs, and did not rely on intrinsic membrane properties. The glutamatergic synaptic activity was sensitive to osmotic challenges and kappa-opioid receptor activation, physiological stimuli known to affect phasic activity. Together, our data thus strongly suggest that phasic activity in magnocellular VP neurons is controlled by glutamatergic synaptic inputs rather than by intrinsic properties.


The transcriptional regulation of miR-21, its multiple transcripts, and their implication in prostate cancer.
MicroRNAs (miRNAs) are a natural part of the most recently discovered and global regulatory pathway known as RNA interference. Functional studies have shown how specific miRNAs can function as tumor suppressors or oncogenes and, correspondingly, deregulated miRNA profiles have been observed in prostate and other cancers. However, the upstream pathways which regulate miRNA expression are only currently being uncovered. The Androgen Receptor (AR) is a nuclear hormone receptor and transcription factor which plays a paramount role in prostate cancer (PCa) pathobiology. We performed high throughput miRNA microarray analysis on two AR-responsive cell lines to identified 16 candidate AR-regulated miRNAs.(1) One of the most androgen-induced candidates was a known oncogenic miRNA, miR-21. In a small study of early grade PCa samples we found that miR-21 levels were frequently elevated in comparison to adjacent normal tissue. This observation was supported in the literature(2,3) and suggests clinical relevance. We found that the activated AR directly interacts with miR-21 regulatory regions, indicating direct transcriptional induction. Furthermore, we provide new reporter studies supporting AR-regulation. Importantly, in functional studies, we found that a modest overexpression of miR-21 enhanced tumor xenograft growth and was sufficient to support androgen-independent proliferation following surgical castration. Thus, our studies suggest a model where miR-21 contributes to androgen-dependent and androgen-independent PCa growth. However, the AR is only one of many reported transcriptional regulators of miR-21. Here we review our recent discoveries and further analyze the reported miR-21 regulatory regions, inhibitory and stimulatory signaling pathways, and primary transcripts.


Measurement of tear secretion in healthy pigeons by using the phenol red thread test.
The objectives of this study were to determine the applicability of the phenol red thread (PRT) test as a new method for the evaluation of tear secretion in healthy male and female pigeons, to establish normal physiologic reference values in these animals, and to compare seasonal variations in these values. Seventy-five pigeons of both sexes, with no ocular abnormalities, were included in the study. The phenol red impregnated thread was inserted into the recessus conjunctiva inferior for measuring tear secretion. After 15 seconds, the thread was removed and the wet portion of the thread was measured (in mm). The mean±sd PRT values were 23.02±2.98 mm/15 seconds and 24.04±2.60 mm/15 seconds for the April and June measurements, respectively. There were significant differences in the PRT between the two months, with a significant increase in June. Mean PRT values for males and females were, respectively, 23.30±3.35 and 22.79±2.65 mm/15 seconds in April, and 24.57±2.41 and 23.61±2.68 mm/15 seconds in June. There was no significant correlation (r=0.075, P>0.05) between bodyweight and tear production in both eyes of the male and female birds in April and June. The results indicated that seasonal variations have an effect on tear production. The PRT was a viable method for the measurement of tear production in pigeons and these measurements could be accepted as reference values for healthy pigeons.


Comparison of the effect of propofol and sevoflurane on the urethral pressure profile in healthy female dogs.
To compare the effects of propofol and sevoflurane on the urethral pressure profile in female dogs. 10 healthy female dogs. Urethral pressure profilometry was performed in awake dogs, during anesthesia with sevoflurane at 1.5, 2.0, and 3.0% end-tidal concentration, and during infusion of propofol at rates of 0.4, 0.8, and 1.2 mg/kg/min. A consistent plane of anesthesia was maintained for each anesthetic protocol. Maximum urethral pressure, maximum urethral closure pressure, functional profile length, and functional area were measured. Mean maximum urethral closure pressure of awake dogs was not significantly different than that of dogs anesthetized with propofol at all infusion rates or with sevoflurane at 1.5 and 2.0% end-tidal concentration. Functional area in awake dogs was significantly higher than in anesthetized dogs. Functional area of dogs during anesthesia with sevoflurane at 3.0% end-tidal concentration was significantly lower than functional area for other anesthetic protocols. Individual differences in the magnitude of effects of propofol and sevoflurane on urethral pressures were observed. Sevoflurane is an alternative to propofol for anesthesia in female dogs undergoing urethral pressure profilometry. Use of these anesthetics at appropriate administration rates should reliably distinguish normal from abnormal maximum urethral closure pressures and functional areas. Titration of anesthetic depth is a critical component of urodynamic testing.


Cochlear gene therapy with ancestral AAV in adult mice: complete transduction of inner hair cells without cochlear dysfunction.
The use of viral vectors for inner ear gene therapy is receiving increased attention for treatment of genetic hearing disorders. Most animal studies to date have injected viral suspensions into neonatal ears, via the round window membrane. Achieving transduction of hair cells, or sensory neurons, throughout the cochlea has proven difficult, and no studies have been able to efficiently transduce sensory cells in adult ears while maintaining normal cochlear function. Here, we show, for the first time, successful transduction of all inner hair cells and the majority of outer hair cells in an adult cochlea via virus injection into the posterior semicircular canal. We used a "designer" AAV, AAV2/Anc80L65, in which the main capsid proteins approximate the ancestral sequence state of AAV1, 2, 8, and 9. Our injections also transduced ~10% of spiral ganglion cells and a much larger fraction of their satellite cells. In the vestibular sensory epithelia, the virus transduced large numbers of hair cells and virtually all the supporting cells, along with close to half of the vestibular ganglion cells. We conclude that this viral vector and this delivery route hold great promise for gene therapy applications in both cochlear and vestibular sense organs.


Identification of structural variation in mouse genomes.
Structural variation is variation in structure of DNA regions affecting DNA sequence length and/or orientation. It generally includes deletions, insertions, copy-number gains, inversions, and transposable elements. Traditionally, the identification of structural variation in genomes has been challenging. However, with the recent advances in high-throughput DNA sequencing and paired-end mapping (PEM) methods, the ability to identify structural variation and their respective association to human diseases has improved considerably. In this review, we describe our current knowledge of structural variation in the mouse, one of the prime model systems for studying human diseases and mammalian biology. We further present the evolutionary implications of structural variation on transposable elements. We conclude with future directions on the study of structural variation in mouse genomes that will increase our understanding of molecular architecture and functional consequences of structural variation.


A strict anaerobic extreme thermophilic hydrogen-producing culture enriched from digested household waste.
The aim of this study was to enrich, characterize and identify strict anaerobic extreme thermophilic hydrogen (H(2)) producers from digested household solid wastes. A strict anaerobic extreme thermophilic H(2) producing bacterial culture was enriched from a lab-scale digester treating household wastes at 70 degrees C. The enriched mixed culture consisted of two rod-shaped bacterial members growing at an optimal temperature of 80 degrees C and an optimal pH 8.1. The culture was able to utilize glucose, galactose, mannose, xylose, arabinose, maltose, sucrose, pyruvate and glycerol as carbon sources. Growth on glucose produced acetate, H(2) and carbon dioxide. Maximal H(2) production rate on glucose was 1.1 mmol l(-1) h(-1) with a maximum H(2) yield of 1.9 mole H(2) per mole glucose. 16S ribosomal DNA clone library analyses showed that the culture members were phylogenetically affiliated to the genera Bacillus and Clostridium. Relative abundance of the culture members, assessed by fluorescence in situ hybridization, were 87 +/- 5% and 13 +/- 5% for Bacillus and Clostridium, respectively. An extreme thermophilic, strict anaerobic, mixed microbial culture with H(2)-producing potential was enriched from digested household wastes. This study provided a culture with a potential to be applied in reactor systems for extreme thermophilic H(2) production from complex organic wastes.


Process evaluation of the Bristol girls dance project.
The Bristol Girls Dance Project was a cluster randomised controlled trial that aimed to increase objectively measured moderate-to-vigorous physical activity (MVPA) levels of Year 7 (age 11-12) girls through a dance-based after-school intervention. The intervention was delivered in nine schools and consisted of up to forty after-school dance sessions. This paper reports on the main findings from the detailed process evaluation that was conducted. Quantitative and qualitative data were collected from intervention schools. Dose and fidelity were reported by dance instructors at every session. Intervention dose was defined as attending two thirds of sessions and was measured by attendance registers. Fidelity to the intervention manual was reported by dance instructors. On four randomly-selected occasions, participants reported their perceived level of exertion and enjoyment. Reasons for non-attendance were self-reported at the end of the intervention. Semi-structured interviews were conducted with all dance instructors who delivered the intervention (n = 10) and school contacts (n = 9) in intervention schools. A focus group was conducted with girls who participated in each intervention school (n = 9). The study did not affect girls' MVPA. An average of 31.7 girls participated in each school, with 9.1 per school receiving the intervention dose. Mean attendance and instructors' fidelity to the intervention manual decreased over time. The decline in attendance was largely attributed to extraneous factors common to after-school activities. Qualitative data suggest that the training and intervention manual were helpful to most instructors. Participant ratings of session enjoyment were high but perceived exertion was low, however, girls found parts of the intervention challenging. The intervention was enjoyed by participants. Attendance at the intervention sessions was low but typical of after-school activities. Participants reported that the intervention brought about numerous health and social benefits and improved their dance-based knowledge and skills. The intervention could be improved by reducing the number of girls allowed to participate in each school and providing longer and more in-depth training to those delivering the intervention. ISRCTN52882523 Registered 25th April 2013.


Chronic herpesvirus reactivation occurs in aging.
The aged immune system is characterized by clonal expansions of CD8+ T cells of which a substantial portion are directed against Epstein-Barr virus (EBV) and cytomegalovirus (CMV). It is unknown if these expansions represent increased viral reactivation or simply reflect an accumulation over time. We investigated herpesvirus reactivation in young and old subjects co-infected with CMV and EBV. Using molecular and serological techniques, we found significant increases in both the frequency and magnitude of EBV and CMV reactivation in elderly subjects. CMV DNA was frequently detected in the urine of elderly subjects; EBV load in peripheral blood was also significantly increased. Notably, EBV DNA in plasma was detected in a majority of the elderly subjects which was supported by frequent transcription of late structural genes. Furthermore, CD8+ T cells specific for EBV structural antigens were detected in samples from the elderly. Samples from our younger control group were negative for EBV DNA in plasma, CMV DNA in urine, expression of structural transcripts, and lacked CD8+ T cells specific for EBV structural antigens. These findings indicate that the aged immune system is no longer able to control EBV and CMV reactivation that could now be characterized as chronic instead of latent.


Subclinical cerebral edema in children with diabetic ketoacidosis randomized to 2 different rehydration protocols.
Previous studies show that vasogenic cerebral edema (CE) occurs during diabetic ketoacidosis (DKA) treatment in children, but the role of intravenous fluids in contributing to CE is unclear. We used magnetic resonance diffusion weighted imaging to quantify subclinical CE in children with DKA randomized to 2 intravenous fluid regimens. Children with DKA were randomized to receive fluids at a more rapid rate (n = 8) or a slower rate (n = 10), with all other aspects of DKA treatment kept identical. Children underwent diffusion weighted imaging 3 to 6 hours and 9 to 12 hours after beginning DKA treatment and after recovery from DKA (≥ 72 hours after beginning treatment). We calculated brain apparent diffusion coefficient (ADC) values as the average of measurements in the basal ganglia, thalamus, frontal white matter, and hippocampus and determined the mean brain ADC value during DKA treatment by averaging data from the 3- to 6-hour and 9- to 12-hour measurements. The difference in mean brain ADC between DKA treatment and postrecovery was used as an index of the severity of CE during DKA treatment. Mean brain ADC values during DKA treatment were significantly higher than postrecovery values, consistent with vasogenic CE (842 ± 38 vs 800 ± 41 × 10(-6) mm(2)/second, P = .002). We did not detect significant differences in ADC elevation in children treated with more rapid versus slower rehydration (β coefficient 0.11 for 1 SD change in ADC, 95% confidence interval: -0.91 to 1.13). ADC changes during DKA treatment (reflective of vasogenic CE) do not appear to be substantially affected by the rate of intravenous fluid administration.


Opposing time trends of peptic ulcer and reflux disease.
Gastritis associated hypochlorhydria may be protective against gastro-oesophageal reflux disease. It was hypothesised that the historic decline in Helicobacter pylori infection resulted in a decline in peptic ulcer and a concomitant risk in reflux disease. To study the time trends of peptic ulcer and reflux disease. Hospitalisation rates were analysed using the computerised database of the US Department of Veterans Affairs from 1970 until 1995. Death rates were calculated from the Vital Statistics of the United States. From 1970 to 1995, hospitalisation rates for gastric and duodenal ulcer, as well as gastric cancer fell, while the hospitalisation rates for gastro-oesophageal reflux disease and those for oesophageal adenocarcinoma rose significantly. Similar time trends were observed with respect to the death rates. There were notable ethnic differences. White subjects incurred higher rates of reflux disease and oesophageal adenocarcinoma and lower rates of gastric ulcer or cancer compared with non-whites. The ethnic variations and the opposing time trends of gastroduodenal versus oesophageal disease are consistent with the hypothesis that the declining infection rates of H pylori in the general population have led to a rise in the occurrence of gastro-oesophageal reflux disease and associated oesophageal adenocarcinoma.


Asthma hospitalizations and mortality in Chicago: an epidemiologic overview.
To characterize the patterns and correlates of asthma hospitalizations and mortality in Chicago. Cross-sectional analysis of discharge data for 1996 and mortality time trend data for the period from 1990 to 1997. The city of Chicago, IL, with Cook County, IL, and US data employed for comparisons. People who were hospitalized with a primary diagnosis of asthma and people whose underlying cause of death was asthma. None. The 1996 asthma hospitalization rate for Chicago was 42.8 per 10,000, more than twice as high as suburban Chicago or US rates. Medicaid patients were overrepresented. Length of stay was longer for older patients and Medicaid patients. Age-adjusted asthma mortality in Chicago was 4.7 times higher in non-Hispanic blacks than in non-Hispanic whites. The black/white asthma mortality ratio is 2.5:1 for the nation overall. Asthma mortality rates for Hispanics in Chicago were between those of non-Hispanic whites and blacks but have almost doubled during this decade. The rising asthma mortality and high asthma hospitalization rates in Chicago constitute a significant public health problem. Comorbidities more common in urban environments, such as substance abuse, may play a unique role in determining the distribution of adverse outcomes within Chicago's population. Asthma hospitalizations and deaths may vary in their risk profiles, and this should be taken into account when developing research and intervention strategies.


The pokeweed antiviral protein specifically inhibits Ty1-directed +1 ribosomal frameshifting and retrotransposition in Saccharomyces cerevisiae.
Programmed ribosomal frameshifting is a molecular mechanism that is used by many RNA viruses to produce Gag-Pol fusion proteins. The efficiency of these frameshift events determines the ratio of viral Gag to Gag-Pol proteins available for viral particle morphogenesis, and changes in ribosomal frameshift efficiencies can severely inhibit virus propagation. Since ribosomal frameshifting occurs during the elongation phase of protein translation, it is reasonable to hypothesize that agents that affect the different steps in this process may also have an impact on programmed ribosomal frameshifting. We examined the molecular mechanisms governing programmed ribosomal frameshifting by using two viruses of the yeast Saccharomyces cerevisiae. Here, we present evidence that pokeweed antiviral protein (PAP), a single-chain ribosomal inhibitory protein that depurinates an adenine residue in the alpha-sarcin loop of 25S rRNA and inhibits translocation, specifically inhibits Ty1-directed +1 ribosomal frameshifting in intact yeast cells and in an in vitro assay system. Using an in vivo assay for Ty1 retrotransposition, we show that PAP specifically inhibits Ty1 retrotransposition, suggesting that Ty1 viral particle morphogenesis is inhibited in infected cells. PAP does not affect programmed -1 ribosomal frameshift efficiencies, nor does it have a noticeable impact on the ability of cells to maintain the M1-dependent killer virus phenotype, suggesting that -1 ribosomal frameshifting does not occur after the peptidyltransferase reaction. These results provide the first evidence that PAP has viral RNA-specific effects in vivo which may be responsible for the mechanism of its antiviral activity.


Detection and management of hypothyroidism following thyroid lobectomy: evaluation of a clinical algorithm.
The objectives of this study were to determine: (1) the incidence permanent hypothyroidism after thyroid lobectomy (TL), (2) whether asymptomatic patients with mildly elevated thyrotropin (TSH) levels can be managed without thyroid hormone replacement, and (3) if the degree of lymphocytic infiltration (LI) and germinal center (GC) formation in the resected thyroid lobe correlates with the development of post-TL hypothyroidism. Subjects undergoing TL between January 2006 and January 2008 at 2 centers were enrolled in the study and thyroid function was followed prospectively based on a previously published algorithm. The histology of each resected thyroid lobe was examined, and the degree of LI and GC was quantified. The study cohort consisted of 117 patients. Early postoperative TSH levels were significantly increased over preoperative levels (P < .001). TSH measured at 6 months to 1 year postoperatively, while still significantly increased over preoperative levels (P < .001), was also significantly reduced (P = .006) compared with early postoperative levels. Of the patients who presented with early postoperative hypothyroidism, 69.2% recovered to normal levels without intervention. The overall incidence of early postoperative hypothyroidism was 21.6%, and permanent hypothyroidism was 7.8%. A high degree of LI and GC correlated with a significantly higher mean TSH level (P = .003). The incidence of hypothyroidism following TL is low, and a significant proportion of individuals who become biochemically hypothyroid will demonstrate only a transient elevation in their TSH levels. As well, individuals with LI, or GC formation, within their resected thyroid lobe may be at increased risk for post-TL hypothyroidism.


Extracolonic findings in patients undergoing abdomino-pelvic CT for suspected colorectal carcinoma in the frail and disabled patient.
The aims of this study were to evaluate the extracolonic findings identified in patients undergoing minimal preparation abdomino-pelvic CT in place of barium enema or colonoscopy for the detection of possible colorectal carcinoma. The CT technique involved helical acquisition (10 mm collimation, 1.5 pitch) following 2 days of preparation with oral contrast medium only. Extracolonic findings were evaluated in the light of subsequent follow-up and accuracy. The evaluation included assessment of the potential contribution of the extracolonic finding(s) to staging the cancer in the subset of patients who had colorectal carcinoma, and to account for the patients' presenting symptoms and signs in the remaining patients. A total of 344 extracolonic findings were detected in 261 CT examinations, from amongst a total of 1077 cases (24%). Extracolonic findings were potentially important in staging in 32 of the 98 (33%) cases subsequently found to have colorectal cancer. There were 284 extracolonic findings amongst the 221 cases who proved not have colorectal cancer. One hundred and twenty-four (44%) of these 284 findings were actively followed up by clinicians, and 33 (12%) ultimately had a surgical intervention. Fifty-six percent (160/284) of the findings were determined to be correct (by further investigation, autopsy, and/or clinical follow-up); the remainder were incorrect or indeterminate (n = 56) or had no follow-up (n = 68). The commonest extracolonic findings were focal liver lesions (found in 42/1077, 4%) and abdominal aortic aneurysms (31/1077, 3%). Twenty-four (24/1077, 2%) previously unknown extracolonic malignancies were detected. Ten percent (106/1077) of the patients had extracolonic findings that could potentially have accounted for their presenting symptoms. CT has the added benefit, compared with colonoscopy and barium enema, of not just evaluating the colon but also of detecting extracolonic abnormalities. Such findings may be useful in staging the cancer, may explain the patient's presenting symptoms, and may detect other potentially serious disorders.


Emotional distress among parent caregivers of adolescents with sickle cell disease: Association with patients and caregivers variables.
Evidence suggests that impairment in caregiver wellbeing can alter the quality of care in children with sickle cell disease. We examined 121 parent caregivers of adolescents with sickle cell disease for emotional distress and disruptions to caregiver lifestyle and interests. Participants were predominantly mothers 92(76%) with mean age, 43.59 (SD = 6.39) years. Four in every ten caregivers had emotional distress, and this was predicted by frequent hospital attendance and disruptions in caregiver lifestyle, relationships, and interests. Psychosocial support to promote resilience and adaptive coping-styles to deal with the stress from unforeseen crises, frequent hospital visits, and lifestyle disruptions are indicated to improve caregiver wellbeing.


In Vivo and In Vitro Hepatoprotective Effects of Geranium koreanum Methanolic Extract via Downregulation of MAPK/Caspase-3 Pathway.
Geranium koreanum (GK) is an indigenous Chinese herbal medicine widely used for the treatment of various inflammation and liver disorders. However, the exact mechanism of action of GK remains unknown. This study aimed to investigate the protective effect and related molecular mechanism of GK on NaAsO2-induced cytotoxicity in HepG2 cells and liver damage in mice. The cytoprotective role of GK was assessed on HepG2 cells using MTT assay. Oxidative stress and lactate dehydrogenase levels were measured with ROS and LDH assay. Histopathology and serum enzymes levels were estimated. The molecular mechanism was evaluated by qPCR and immunoblotting to ensure the hepatoprotective role of GK against NaAsO2 intoxication in mice. We found cotreatment with GK significantly attenuated NaAsO2-induced cell viability loss, intracellular ROS, and LDH release. Hepatic histopathology and serum biochemical parameters, ALT, and AST were notably improved by cotreatment with GK. Beside, GK markedly altered both mRNA and protein expression level of MAPK. The proapoptotic and antiapoptotic protein Bax/Bcl-2 ratio was significantly regulated by GK. Moreover, GK remarkably suppressed the postapoptotic transcription protein cleaved caspase-3 expression. The present study reveals that GK possesses hepatoprotective activity which is probably involved in the modulation of the MAPK/caspase-3 pathway.


Abnormal hepatic sinusoidal bile acid transport in an Amish kindred is not linked to FIC1 and is improved by ursodiol.
The mechanism for abnormal hepatic bile acid transport was investigated in an 18-month-old Amish boy who presented with pruritus, poor growth, and severe bleeding episodes. Serum bilirubin, gamma-glutamyltranspeptidase, and cholesterol levels were normal, but prothrombin time and partial thromboplastin time were prolonged and bone alkaline phosphatase level was elevated. Cholic acid plus chenodeoxycholic acid levels measured by capillary gas-chromatography were 32 times higher than control in serum (34.7 vs. 1.1+/-0.4 microg/dL) but were not detected in liver and were reduced in gallbladder bile. Treatment with ursodiol, a more hydrophilic bile acid, improved pruritus, produced 37% weight gain, and after 2 years reduced serum primary bile acid concentrations about 85%, while accounting for 71% of serum and 24% of biliary bile acid conjugates. On ursodiol therapy, hepatic bile acid synthesis was enhanced 2-fold compared with controls, and microscopy revealed chronic hepatitis without cholestasis. Three younger sisters with elevated serum bile acids responded positively to ursodiol. Microsatellite markers for the FIC1 (gene for Byler's disease) region in these 4 children were inconsistent with linkage to FIC1. Conjugated cholic acid and chenodeoxycholic acid were synthesized in the liver and secreted into bile but could not reenter the liver from portal blood and accumulated in serum. In contrast, unconjugated ursodiol entered the liver and was conjugated and secreted into bile. Thus, the enterohepatic circulation of all conjugated bile acids was interrupted at the hepatic sinusoidal basolateral membrane. Unconjugated ursodiol bypassed the hepatic uptake block to enlarge the biliary and intestinal bile acid pools. A mutation in FIC1 recognized among the Amish and linkage of the disorder to FIC1 were excluded.


Decreased GAP-43 accumulation in neurite tips of cultured hippocampal neurons by acrylamide.
The critical axonopathy- producing effect of acrylamide (ACR) is hypothesized to be an interruption of fast vesicular transport resulting in a deficiency of distal axonal proteins. This study used the accumulation of growth associated protein- 43 (GAP-43) in neurite tips of cultured hippocampal neurons as a model to examine the effect of an ACR block of fast transport on the eventual protein concentration within the distal axon. Twenty-four hour incubation with ACR produced a dose-dependent reduction with 0.5mM and 1.0mM concentrations in the percent of neurites with a terminal GAP-43 accumulation. No change in cell size (area), cell body GAP-43 fluorescence, distal neurite fluorescence, absolute tip fluorescence or neurite length were observed. Another known inhibitor of transport, colchicine (COL) also caused a significant decrease in the percent of neurites with a GAP-43 accumulation; propionamide (PA), a non-neurotoxic analogue of ACR, had no effect on protein accumulation. These findings support the hypothesis that the disruption of fast transport by ACR can lead to depletion of vital proteins in the distal axon.


Short daily haemodialysis: survival in 415 patients treated for 1006 patient-years.
Survival statistics for daily haemodialysis are lacking as most centres providing this have treated only a small number of patients for short observation times. We pooled our 23-year, 1006-patient-year, five-centre experience of 415 patients treated by short daily haemodialysis. One hundred and fifty patients were treated in-centre, most because of medical complications and 265 by home or self-care haemodialysis. Patients were on daily haemodialysis for 29 +/- 31 (0-272) months. Forty-two percent had primary and 31% had secondary renal failure. Treatment time was 136 +/- 35 min, frequency 5.8 +/- 0.5 times/week and weekly stdKt/V 2.7 +/- 0.55. Eighty-five patients (20%) died; 5-year cumulative survival was 68 +/- 4.1% and 10-year survival was 42 +/- 9%. Age, secondary renal failure and in-centre dialysis were associated with mortality, while gender, frequency of dialysis (5, 6 or 7 per week), continent, country and blood access were not. Survival was compared with matched patients from the USRDS 2005 Data Report using the standardized mortality ratio and cumulative survival curves. Both comparisons showed that the survival of the daily haemodialysis patients was 2-3 times higher and the predicted 50% survival time 2.3-10.9 years longer than that of the matched US haemodialysis patients. Survival of patients dialyzing daily at home was similar to that of age-matched recipients of deceased donor renal transplants. Survival of patients on short daily haemodialysis was 2-3 times better than that of matched three times weekly haemodialysis patients reported by the USRDS.


An Anomalous Noise Events Detector for Dynamic Road Traffic Noise Mapping in Real-Life Urban and Suburban Environments.
One of the main aspects affecting the quality of life of people living in urban and suburban areas is their continued exposure to high Road Traffic Noise (RTN) levels. Until now, noise measurements in cities have been performed by professionals, recording data in certain locations to build a noise map afterwards. However, the deployment of Wireless Acoustic Sensor Networks (WASN) has enabled automatic noise mapping in smart cities. In order to obtain a reliable picture of the RTN levels affecting citizens, Anomalous Noise Events (ANE) unrelated to road traffic should be removed from the noise map computation. To this aim, this paper introduces an Anomalous Noise Event Detector (ANED) designed to differentiate between RTN and ANE in real time within a predefined interval running on the distributed low-cost acoustic sensors of a WASN. The proposed ANED follows a two-class audio event detection and classification approach, instead of multi-class or one-class classification schemes, taking advantage of the collection of representative acoustic data in real-life environments. The experiments conducted within the DYNAMAP project, implemented on ARM-based acoustic sensors, show the feasibility of the proposal both in terms of computational cost and classification performance using standard Mel cepstral coefficients and Gaussian Mixture Models (GMM). The two-class GMM core classifier relatively improves the baseline universal GMM one-class classifier F1 measure by 18.7% and 31.8% for suburban and urban environments, respectively, within the 1-s integration interval. Nevertheless, according to the results, the classification performance of the current ANED implementation still has room for improvement.


Computational analysis of the yeast proteome: understanding and exploiting functional specificity in genomic data.
Modern experimental techniques have produced a wealth of high-throughput data that has enabled the ongoing genomic revolution. As the field continues to integrate experimental and computational analyzes of this data, it is essential that performance evaluations of high-throughput results be carried out in a consistent and biologically informative manner. Here, we present an overview of evaluation techniques for high-throughput experimental data and computational methods, and we discuss a number of potential pitfalls in this process. These primarily involve the biological diversity of genomic data, which can be masked or misrepresented in overly simplified global evaluations. We describe systems for preserving information about biological context during dataset evaluation, which can help to ensure that multiple different evaluations are more directly comparable. This biological variety in high-throughput data can also be taken advantage of computationally through data integration and process specificity to produce richer systems-level predictions of cellular function. An awareness of these considerations can greatly improve the evaluation and analysis of any high-throughput experimental dataset.


Intentions of burial: mourning, politics, and memorials following the massacre at Srebrenica.
In the aftermath of tragedy, memorials and grave sites are frequently constructed. Research needs to pay attention to the goals of mourners for such places. The 1995 massacre at Srebrenica in Bosnia-Herzegovina left over 7,000 Muslim men missing and presumed dead. Five years later, families fought to bury the remains that had been collected. Qualitative interviews with 67 people, including survivors and key informants were completed over the summer of 2000. This article examines the intentions of the burial from the perspective of two groups of family members who lost loved ones: survivors who remain politically isolated and those involved in advocacy or political organizations. For the politically isolated, burial was important in reshaping individual mourning after mass death and in the face of unidentified remains. For those involved in advocacy groups, burial was a means of recognizing the past massacre, understanding the massacre's impact on current political divisions, and shaping future directions. Both mourning and political functions are elaborated on in the setting of Bosnian culture and society.


Evaluating a Commonly Used Tool for Measuring Sport Specialization in Young Athletes.
Sport specialization has been defined as year-round intensive training in a single sport to the exclusion of other sports. A commonly used survey tool created by Jayanthi et al, which classifies athletes as having a low, moderate, or high level of specialization, categorizes only athletes answering yes to "Have you quit other sports to focus on a main sport?" as highly specialized. We hypothesized that a measureable number of year-round, single-sport athletes have never played other sports and, therefore, may be inaccurately classified as moderately specialized when using this tool, even though most experts would agree they should be viewed as highly specialized. To determine the proportion of athletes misclassified as moderately rather than highly specialized because they never played a previous sport. Cross-sectional study. Hospital-based pediatric outpatient sports medicine clinic. Injured athletes aged 12 to 17 years who presented to the clinic between 2015 and 2017 and completed a sports-participation survey (n = 917). Sport-specialization level. Of 917 participants, 299 (32.6%) played a single sport more than 8 months per year, and 208/299 (69.6%) had previously quit other sports (highly specialized), whereas 91 (30.4%) had never played other sports (highly specialized and misclassified as moderate). Individual-sport athletes had a 2.03 times greater risk of being highly specialized and misclassified as moderate than team-sport athletes (relative risk = 2.03 [95% confidence interval = 1.37, 3.00]). Females had a 1.70 times greater risk of being misclassified as moderately specialized than males (relative risk 1.70 [95% confidence interval = 1.07, 2.70]). Of the 3 sports with the largest number of athletes, artistic gymnastics had the highest proportion (51.2%) who had never played other sports. The commonly used specialization survey misclassified a substantial number of highly specialized athletes as moderately specialized. Researchers should consider adding a fourth survey question, "Have you only ever played 1 sport?" to identify and better study this unique subset of misclassified athletes.


Human immunodeficiency virus type 1 nucleocapsid protein promotes efficient strand transfer and specific viral DNA synthesis by inhibiting TAR-dependent self-priming from minus-strand strong-stop DNA.
During the first strand transfer in reverse transcription, minus-strand strong-stop DNA [(-) SSDNA] is annealed to the 3' end of the acceptor RNA in a reaction mediated by base-pairing between terminal repeat sequences in the RNA and their complement in the DNA. The large stem-loop structure in the repeat region known as TAR could interfere with this annealing reaction. We have developed an in vitro human immunodeficiency virus type 1 (HIV-1) system to investigate the effect of TAR on strand transfer. Mutational analysis demonstrates that the presence of TAR in the donor and acceptor templates inhibits strand transfer and is correlated with extensive synthesis of heterogeneous DNAs formed by self-priming from (-) SSDNA. These DNAs are not precursors to the transfer product. Interestingly, products of self-priming are not detected in HIV-1 endogenous reactions; this suggests that virions contain a component which prevents self-priming. Our results show that the viral nucleocapsid protein (NC), which can destabilize secondary structures, drastically reduces self-priming and dramatically increases the efficiency of strand transfer. In addition, the data suggest that the ability to eliminate self-priming is a general property of NC which is manifested during reverse transcriptase pausing at sites of secondary structure in the template. We conclude that this activity of NC is critical for achieving highly efficient and specific viral DNA synthesis. Our findings raise the possibility that inactivation of NC could provide a new approach for targeting reverse transcription in anti-HIV therapy.


From sea to shining sea: making collaborative rural research work.
Rural researchers collaborate on many levels to collect and analyze data, develop research reports and disseminate findings. While this collaboration is critical, there is a dearth of literature about research team collaboration within all stages of the research process. The purpose of this article is to discuss the research experience of 10 rural researchers scattered across Canada who participated in the study, Health Research: Accessible, Applicable and Useable for Rural Communities and Practitioners. Using focused ethnography, one aim of this study was to discover how research is utilized in rural and remote settings. The necessity of establishing networks to collect and manage data, and jointly analyze 72 qualitative transcripts from different geographical sites led to innovations and unexpected lessons learned. The research design provided significant opportunities to mentor undergraduate, masters and doctoral nursing students and to enhance the development of newly graduated doctoral nurses. These opportunities are crucial in the development of new researchers and in creating ongoing interest in rural health research. In this article, we discuss how the research process evolved, the mentoring process used, the barriers identified related to collaboration across vast distances, and the strategies employed to enhance the study's trustworthiness. We also consider the advantages and challenges of using Elluminate, a web application, as an interactive forum for this qualitative health research.