Presence of different arbuscular mycorrhizal infection patterns in roots of Lotus glaber plants growing in the Salado River basin.
Morphological types of arbuscular mycorrhizal (AM) fungi associated with Lotus glaber in sodic soils of the Salado River basin were studied. At least eight colonization patterns (IP) of AM fungi in roots of L. glaber were observed after 30 plants were analyzed. Arum- and Paris-type infection were found in the same plant species. This result supports the idea that AM morphology is not solely under plant control, but is also influenced by fungal identity. One infection pattern, presumably corresponding to Glomus intraradices, and a second, possibly assignable to Glomus tenue, were the most commonly found. Our results reinforce previous suggestions that G. intraradices is well adapted to sodic-saline conditions and may play a role in the resistance of L. glaber to these soils.


Change of central venous catheter dressings twice a week is superior to once a week in patients with haematological malignancies.
Thirty-two consecutive patients with haematological disorders, in need of a permanent central venous catheter (CVC) were randomly allocated to have their CVC bandages (Tegaderm) changed once (OAW, n = 20) or twice (TAW, n = 19) a week. The two randomization arms were balanced in respect of age, sex, and underlying disease. The exit site of the CVC was inspected daily through the transparent bandage and erythema was noted. If severe erythema occurred, daily wet gauze dressings were applied. Samples for bacterial cultures were taken from the exit site of the CVC at every change of bandages. There was no difference in complications leading to removal of the CVC between the two groups (7/20 OAW vs. 7/19 TAW) or in CVC survival-time (P = 0.4). However, the OAW group had more positive CVC tip cultures (OAW 11/14 vs. TAW 2/9; P < 0.05) and a tendency to: (i) more extra dressings (P = 0.08); (ii) more cultures from the exit skin site showing high numbers of colony forming units (P = 0.07); (iii) shorter time to first exit site infection (P = 0.09); and (iv) more Gram-positive septicaemias (P = 0.08). Both clinical and bacteriological data in this study indicate that changing transparent polyurethane CVC bandages twice a week is superior to once a week.


Topical glyceryltrinitrate causes measurable penile arterial dilation in impotent men.
The current understanding of the intracavernous changes that cause or accompany penile erections has encouraged the use of vasodilators as therapy for erectile dysfunction. An established vasodilator, glyceryltrinitrate, was selected for in vivo study because of its rapid transdermal absorption. Color coded duplex ultrasound was used to assess penile vascular response. In a large group of men with erectile dysfunction significant dilation was noted in response to a small amount of nitroglycerine paste applied to the penis. There is a measurable vasodilatory response that can be induced by synthetic nitrates in penile tissue in impotent men.


Identification of a pH sensor in Influenza hemagglutinin using X-ray crystallography.
Hemagglutnin (HA) mediates entry of influenza virus through a series of conformational changes triggered by the low pH of the endosome. The residue or combination of residues acting as pH sensors has not yet been fully elucidated. In this work, we assay pH effects on the structure of H5 HA by soaking HA crystallized at pH 6.5 in a series of buffers with lower pH, mimicking the conditions of the endosome. We find that HA1-H38, which is conserved in Group 1 HA, undergoes a striking change in side chain conformation, which we attribute to its protonation and cation-cation repulsion with conserved HA1-H18. This work suggests that x-ray crystallography can be applied for studying small-scale pH-induced conformational changes providing valuable information on the location of pH sensors in HA. Importantly, the observed change in HA1-H38 conformation is further evidence that the pH-induced conformational changes of HA are the result of a series of protonation events to conserved and non-conserved pH sensors.


Historical trends of survival among widows and widowers.
One of the most consistent findings in social demography is that recently widowed individuals, male or female, have higher rates of mortality than comparable married persons. These results are based generally on contemporary studies in developed nations where life expectancy is high. Because of data limitations, there are few studies available to determine whether these findings also occur when mortality rates were higher. This study uses the Utah Population Database that was developed from extensive family genealogies and now linked to Utah death certificates. These data make it possible to employ life course analysis of four marriage cohorts extending from 1860 through 1904 with mortality follow-up to 1990. This approach is used to compare mortality risks of widowed males and females relative to comparable married individuals. Covariates included in the study are remarriage, as well as religion and number of children ever born; these are all hypothesized to have protective effects on mortality risks for widowed men and women. Analysis of these data indicates that there are significant differences in the mortality risk for widowed men and women, and it is widowed men who have an excess risk of dying in every cohort and nearly every age. A consistent pattern of excess mortality in the comparison of married and widowed women was not observed. There are significant female and male differences in the effect of religion which was treated as a proxy for life style and social support: however, remarriage as a proxy for social support has similar protective effects on the surviving spouse.


Biosynthesis of the C-terminal amide in peptide hormones.
Recent developments in the study of peptide amidation are reviewed. The main areas covered are assay procedures, purification of amidating enzymes, co-factors and regulation, mechanism and specificity of the amidating reaction, and multiple forms of the amidating enzyme and glycosylation. Discussion is presented on aspects that are poorly understood and new areas open to investigation are indicated.


Local excision after neoadjuvant chemoradiotherapy versus total mesorectal excision: a case-matched study in 110 selected high-risk patients with rectal cancer.
The aim of this comparative study was to report a 10-year experience of an organ preservation strategy by local excision (LE) in selected high-risk patients (aged patients and/or patients with severe comorbidity and/or indication for abdominoperineal excision) versus total mesorectal excision (TME) after neoadjuvant radiochemotherapy (RCT) for patients with locally advanced (T3-T4 and/or N+) low and mid rectal cancer with suspicion of complete tumour response (CTR) or near-CTR. Thirty-nine patients with rectal cancer who underwent LE after RCT for suspicion of CTR were matched to 71 patients who underwent TME according to body mass index, gender, tumour location and ypTNM stage. Operative, oncological and functional results were compared between groups. In the LE group, ypT0, ypTis or ypT1N0R0 were noted in 28/39 (72%). Overall morbidity was observed in 10/39 (26%) in LE vs 46/71 in the TME group (65%) (P = 0.001). Severe morbidity (Clavien-Dindo ≥ 3) was noted in 1/39 patients from the LE group (3%) vs 3/71 (4%) from the TME group (P = 1.000). After a mean follow-up of 63 ± 4 months (range 56-70 months), local recurrence was noted in 2/39 (5%) from the LE group vs 2/71 (3%) from the TME group (P = 0.601). Definitive stoma was noted in 2/39 (6%) from the LE group vs 8/71 (12%) from the TME group (P = 0.489). Major low anterior resection syndrome was noted in 5/23 (22%) from LE group vs 11/33 (33%) from the TME group (P = 0.042). The accuracy of response prediction after RCT was 72% after LE. In high-risk patients, LE represents a safe alternative to TME with better functional results and the same long-term oncological outcome.


Mountaineering and rock-climbing injuries in US national parks.
Mountaineering and rock climbing have become increasingly popular in recent years and involve an estimated 100,000 participants; accordingly, the number of climbing-related injuries has also increased. We analyzed 127 climbing-related injuries reported to the US National Park Service in 1981 and 1982, 36 (28%) of which were fatal. Falls accounted for 75% of all climbing-related injuries; median length of fall was 91 m for fatal injuries and 9 m for nonfatal injuries. The majority of injuries (69%) occurred while ascending. Falls on snow or ice were longer than falls on rock, and injuries on snow or ice were more likely to be fatal. We discuss considerations and strategies for the prevention of climbing-related injuries. A new conceptual model suggests that the methods of traditional mountaineering safety programs may be of limited efficacy in further reducing the number of climbing-related injuries.


Ooplasmic transfer in human oocytes: efficacy and concerns in assisted reproduction.
Ooplasmic transfer (OT) technique or cytoplasmic transfer is an emerging technique with relative success, having a significant status in assisted reproduction. This technique had effectively paved the way to about 30 healthy births worldwide. Though OT has long been invented, proper evaluation of the efficacy and risks associated with this critical technique has not been explored properly until today. This review thereby put emphasis upon the applications, efficacy and adverse effects of OT techniques in human. Available reports published between January 1982 and August 2017 has been reviewed and the impact of OT on assisted reproduction was evaluated. The results consisted of an update on the efficacy and concerns of OT, the debate on mitochondrial heteroplasmy, apoptosis, and risk of genetic and epigenetic alteration. The application of OT technique in humans demands more clarity and further development of this technique may successfully prove its utility as an effective treatment for oocyte incompetence.


Learning probabilistic phenotypes from heterogeneous EHR data.
We present the Unsupervised Phenome Model (UPhenome), a probabilistic graphical model for large-scale discovery of computational models of disease, or phenotypes. We tackle this challenge through the joint modeling of a large set of diseases and a large set of clinical observations. The observations are drawn directly from heterogeneous patient record data (notes, laboratory tests, medications, and diagnosis codes), and the diseases are modeled in an unsupervised fashion. We apply UPhenome to two qualitatively different mixtures of patients and diseases: records of extremely sick patients in the intensive care unit with constant monitoring, and records of outpatients regularly followed by care providers over multiple years. We demonstrate that the UPhenome model can learn from these different care settings, without any additional adaptation. Our experiments show that (i) the learned phenotypes combine the heterogeneous data types more coherently than baseline LDA-based phenotypes; (ii) they each represent single diseases rather than a mix of diseases more often than the baseline ones; and (iii) when applied to unseen patient records, they are correlated with the patients' ground-truth disorders. Code for training, inference, and quantitative evaluation is made available to the research community.


Hospital-based herpes zoster diagnoses in Denmark: rate, patient characteristics, and all-cause mortality.
Herpes zoster (HZ) may result in severe complications requiring hospital treatment, particularly in patients with comorbidity. Nevertheless, data on HZ from nationwide population-based hospital registries are sparse. We conducted a cohort study describing first-time hospital-based (inpatient, outpatient, and emergency room) HZ diagnoses in the Danish National Patient Registry, 1994-2012. We computed the diagnosis rate; prevalence of demographic characteristics, comorbidities, and complications; length of hospital stay; and standardized mortality ratios (SMRs) using the Danish population as reference. We classified comorbidity using the Charlson Comorbidity Index (CCI) scoring system and categorized patients in groups of no (score 0), moderate (score 1), severe (score 2), and very severe comorbidity (score ≥3). In addition, we computed the prevalence of certain conditions associated with immune dysregulation (stem cell or bone marrow transplantation, solid organ transplantation, HIV infection, primary immunodeficiency, any cancer, and autoimmune diseases). The diagnosis rate increased almost exponentially from 6 to 91.9 per 100,000 person-years between age 50 and ≥90 years. The age-standardized rate was stable throughout the study period. The median length of hospital stay was 4 days (interquartile range: 1-8 days) for inpatients with HZ as the main reason for admission. According to the CCI, 44.3 % of patients had no comorbidity, 17.3 % moderate comorbidity, 17.4 % severe comorbidity, and 21.0 % very severe comorbidity. Comorbidities involving immune dysregulation, such as malignant (21 %) and autoimmune diseases (17 %), were particularly prevalent. Thirty percent had neurological, ophthalmic, or other complications. HZ was associated with increased all-cause mortality overall (SMR 1.8, 95 % CI: 1.7-1.8), but not in analyses restricted to patients without comorbidity (SMR 1.0, 95 % CI: 0.9-1.0). This study provides estimates of the epidemiology of hospital-based (severe) HZ. The diagnosis rate increased substantially with age. Complications and comorbidities were prevalent, likely resulting in increased mortality.


Neutral sphingomyelinase 2 deficiency increases hyaluronan synthesis by up-regulation of Hyaluronan synthase 2 through decreased ceramide production and activation of Akt.
Fibroblasts from the fro/fro mouse, with a deletion in the Smpd3 gene coding for the active site of neutral sphingomyelinase 2 (NSMase2), secreted increased amounts of hyaluronan (HA). This was reversed by transfection with the Smpd3 gene, suggesting a connection between sphingolipid and glycosaminoglycan metabolism. The deficiency of NSMase2 resulted in storage of sphingomyelin (SM) and cholesterol with a 50% reduction in ceramides (Cer). RT-PCR and Western blot analysis showed that increased HA secretion resulted from increased hyaluronan synthase 2 (HAS2) activity localized to sphingolipid-enriched lipid rafts. Although cholesterol levels were also elevated in lipid rafts from mouse fibroblasts deficient in lysosomal acid SMase activity (deletion of the Smpd1(-/-) gene), there was no increase in HA secretion. We then showed that in fro/fro fibroblasts, the reduced ceramide was associated with decreased phosphorylation of protein phosphatase 2A (PP2A) and increased phosphorylation of its substrate Akt-p, together with PI3K, PDK1, mTOR (mammalian target of rapamycin), and p70S6K, although PTEN was unaffected. Exogenous ceramide, as well as inhibitors of Akt (Akt inhibitor VIII), PI 3-kinase (LY294002 and wortmannin), and mTOR (rapamycin) reduced secretion of HA, whereas the NSMase2 inhibitor GW4869 increased HA synthesis and secretion. We propose that NSMase2/Cer are the key mediators of the regulation of HA synthesis, via microdomains and the Akt/mTOR pathway.


Preoperative CA 15-3 concentrations predict outcome of patients with breast carcinoma.
CA 15-3 is a breast-associated mucin that is elevated in the majority of breast carcinoma patients with distant metastases. Currently, the main application of this marker is in monitoring and detecting recurrences in patients with diagnosed breast carcinoma. Preoperative serum concentrations (prior to excision of the primary tumor) of CA 15-3 were measured in 368 patients undergoing potentially curative surgical treatment for early breast carcinoma. These results were compared with prospectively recorded clinicopathologic characteristics and patient outcome data. A weak but significant positive association was found between CA 15-3 concentrations and both tumor stage and the number of involved axillary lymph nodes but not between CA 15-3 concentrations and estrogen receptor status. Patients with high concentrations of CA 15-3 had a significantly worse prognosis than patients with low concentrations. Using an optimum cutoff value of 30.38 U/mL, the probability of disease free survival at 5 years was 44% in patients with high CA 15-3 levels compared with 65% in patients with low CA 15-3 levels (P = 0.002, Mantel-Cox log rank test). The corresponding probabilities for overall survival were 67% and 83%, respectively (P < 0.001). The association of preoperative CA 15-3 levels with outcome was maintained in multivariate survival analysis and was not explained by the association between CA 15-3 and tumor size or lymph node burden. The relation between CA 15-3 and outcome also was found within some patient subgroups identified by traditional prognostic factors (axillary lymph node positive patients, patients with primary tumors >2 cm in greatest dimension, and patients with estrogen receptor positive tumors). Preoperative serum concentrations of CA 15-3 appear to have a significant relation to outcome in patients with early breast carcinoma and may have a role in the rational selection of patients for appropriate adjuvant treatments. To the authors' knowledge, CA 15-3 thus is one of the first circulating markers shown to be an independent prognostic indicator in patients with breast carcinoma.


Graft dislocation and hypotony after Descemet's stripping automated endothelial keratoplasty in patients with previous glaucoma surgery.
To determine if patients with prior glaucoma surgery experience higher rates of postoperative graft dislocation after Descemet's stripping automated endothelial keratoplasty (DSAEK) and to determine if postoperative hypotony may be a risk factor in these patients. Retrospective, comparative analysis of an interventional case series. Eight hundred fifty-four eyes (67 eyes with prior glaucoma surgery and 787 controls) from 582 patients who underwent DSAEK at 1 institution between January 2005 and April 2011. Groups were compared with regard to preoperative, intraoperative, and postoperative parameters. Continuous variables were compared using the independent samples t test or Mann-Whitney U test. Categorical variables were compared using the chi-square test or Fisher exact test. Frequencies of postoperative graft dislocation and postoperative hypotony. Study eyes before surgery differed from control eyes with regard to corneal thickness (768 vs. 655 μm; P<0.001) and intraocular pressure (13 vs. 16 mmHg; P<0.001). Postoperative graft dislocation occurred significantly more frequently in study eyes compared with control eyes (9% vs. 2%; P = 0.008). Among eyes in which dislocation occurred, postoperative hypotony was present in 5 study eyes (83%) and 0 control eyes. Previous glaucoma surgery was associated with a significantly increased rate of graft dislocation compared with control eyes. Dislocation was related strongly to postoperative hypotony in eyes with prior glaucoma surgery. Proprietary or commercial disclosure may be found after the references.


What affects annual changes in traffic safety? A macroscopic perspective in Virginia.
Virginia saw a 20% reduction in traffic fatalities in 2008, an unprecedented annual reduction since 1950, and safety stakeholders in Virginia were intrigued about what caused such large a reduction and more generally what affects traffic safety from a macroscopic perspective. This study attempted to find factors associated with such a reduction using historical data of Virginia. Specifically, the study related 18 factors to seven traffic safety measures. In terms of annual changes, the study found that typical crash exposures were not generally associated with the seven measures, while two economic indicators (unemployment rate and U.S. Consumer Price Index [CPI]) were strongly associated with most of them. Annual changes in the CPI and unemployment rate account for about half of the annual changes in total and fatal crash counts, respectively. On average, a 1 point increase in CPI and a 1% increase in the unemployment rate are associated with about 2,500 fewer traffic crashes and about 40 fewer fatal crashes annually in Virginia, respectively.


Synaptic Transmission from Somatostatin-expressing Interneurons to Excitatory Neurons Mediated by α5-subunit-containing GABAA Receptors in the Developing Visual Cortex.
Dendrite-targeting somatostatin-expressing interneurons (SST-INs) powerfully control signal integration and synaptic plasticity in pyramidal dendrites during cortical development. We previously showed that synaptic transmission from SST-INs to pyramidal cells (PCs) (SST-IN → PC) in the mouse visual cortex suddenly declined at around the second postnatal week. However, it is unclear what specific postsynaptic mechanisms underlie this developmental change. Using multiple whole-cell patch-clamp recordings, we found that application of an α5-GABAA receptor-selective inverse agonist, alpha5IA, significantly weakened SST-IN → PC unitary inhibitory postsynaptic currents (uIPSCs) in layer 2/3 of the mouse visual cortex, but had no effect on uIPSCs from SST-INs to other types of interneurons. The extent of alpha5IA-induced reduction of SST-IN → PC synaptic transmission was significantly larger at postnatal days 11-13 (P11-13) than P14-17. Moreover, α5-subunit-containing GABAA receptors (α5-GABAARs)-mediated uIPSCs had slow rise and decay kinetics. Apart from pharmacological test, we observed that SST-IN → PC synapses did indeed contain α5-GABAARs by immunogold labeling for electron microscopy. More importantly, coinciding with the weakening of SST-IN → PC synaptic transmission, the number of α5-GABAAR particles in SST-IN → PC synapses significantly decreased at around the second postnatal week. Together, these data indicate that α5-GABAARs are involved in synaptic transmission from SST-INs to PCs in the neocortex, and are significantly diminished around the second postnatal week.


Degradation of oxytetracycline by iron-manganese modified industrial lignin-based biochar activated peroxy-disulfate: Pathway and mechanistic analysis.
In this study, high-performance Fe-Mn-modified industrial lignin-based biochar (FMBC) was successfully prepared to facilitate the efficient degradation of oxytetracycline by its driven sulfate radical-based advanced oxidation process with 90% degradation within 30 min. The results showed that oxygenated functional groups (e. g. hydroxyl, carbonyl, etc.) in industrial lignin-based biochar, the synergistic effect of transition metals Fe and Mn, and defective structures were the active sites for activation of peroxy-disulfate. SO4·- produced during the degradation process assumed a key function. Significantly, 38 intermediates were innovatively proposed for the first time in the system, and oxytetracycline was degraded in 7 ways, including deamidation, demethylation, hydroxylation, secondary alcohol oxidation, ring opening, dehydration, and carbonylation. A new perspective on the application of industrial lignin in the advanced oxidative degradation of organic pollutants was provided by this study.


[Analysis of costs and cost-effectiveness of preferred GESIDA/National AIDS Plan regimens for initial antiretroviral therapy in human immunodeficiency virus infected adult patients in 2013].
The GESIDA and National AIDS Plan panel of experts have proposed "preferred regimens" of antiretroviral treatment (ART) as initial therapy in HIV infected patients for 2013. The objective of this study is to evaluate the costs and effectiveness of initiating treatment with these "preferred regimens". An economic assessment of costs and effectiveness (cost/effectiveness) was performed using decision tree analysis models. Effectiveness was defined as the probability of having viral load <50copies/mL at week48, in an intention-to-treat analysis. Cost of initiating treatment with an ART regime was defined as the costs of ART and its consequences (adverse effects, changes of ART regime and drug resistance analyses) during the first 48weeks. The perspective of the analysis is that of the National Health System was applied, only taking into account differential direct costs: ART (official prices), management of adverse effects, resistance studies, and determination of HLA B*5701. The setting is Spain and the costs are those of 2013. A sensitivity deterministic analysis was performed, constructing three scenarios for each regimen: baseline, most favourable, and most unfavourable cases. In the baseline case scenario, the cost of initiating treatment ranges from 6,747euros for TDF/FTC+NVP to 12,059euros for TDF/FTC+RAL. The effectiveness ranges between 0.66 for ABC/3TC+LPV/r and ABC/3TC+ATV/r, and 0.87 for TDF/FTC+RAL and ABC/3TC+RAL. Effectiveness, in terms of cost/effectiveness, varies between 8,396euros and 13,930euros per responder at 48weeks, for TDF/FTC/RPV and TDF/FTC+RAL, respectively. Taking ART at official prices, the most effective regimen was TDF/FTC/RPV, followed by the rest of non-nucleoside containing regimens. The sensitivity analysis confirms the robustness of these findings.


Systematic analysis of ear-nose-throat malpractice complaints may be beneficial for patient safety.
The analysis of malpractice complaints can provide valuable information on patient safety. This study offers a detailed examination of the backgrounds concerning reasons and outcomes of ear, nose and throat (ENT) malpractice complaints handled by the National Board of Patients' Complaints (NBPC), 1998-2008. All NBPC decisions related to the ENT specialty from 1998-2008 in Denmark were analysed. Deficient medical recording was criticized even though the complainant did not mention this in the complaint. The juridical theme of medical error was the most complained about, but the least criticized compared with six other juridical themes. In contrast, retained packaging, wrong-site surgery, laser burns and peeling injuries were medical errors with high rates of criticism. Complaints concerning accidental surgical complications did not lead to criticism. Tonsillectomy was the most frequently alleged ENT operation in our study data. Thorough preoperative information about potential surgical complications undersigned by the patient and the newly introduced safe surgery procedures are likely to reduce both faults and complaints. Malpractice complaints have not hitherto been used as a learning instrument. We propose a systematic registration of these incidents to enable more learning in the future. The new act on the handling of malpractice complaints may make it easier to avoid malpractice and to learn from malpractice complaints and may thus improve patient safety in Denmark. not relevant. not relevant.


RNA-binding motif protein 39 (RBM39): An emerging cancer target.
RNA-binding motif protein 39 (RBM39) is an RNA-binding protein involved in transcriptional co-regulation and alternative RNA splicing. Recent studies have revealed that RBM39 is the unexpected target of aryl sulphonamides, which act as molecular glues between RBM39 and the DCAF15-associated E3 ubiquitin ligase complex leading to selective degradation of the target. Loss of RBM39 leads to aberrant splicing events and differential gene expression, thereby inhibiting cell cycle progression and causing tumour regression in a number of preclinical models. Many clinical studies have shown that aryl sulphonamides were well tolerated, but their clinical performance was limited due to an insufficient understanding of the target, RBM39 biology and a lack of predictive biomarkers. This review summarises the current knowledge of RBM39 function and discusses the therapeutic potential of this spliceosome target in cancer therapy. LINKED ARTICLES: This article is part of a themed issue on New avenues in cancer prevention and treatment (BJP 75th Anniversary). To view the other articles in this section visit http://onlinelibrary.wiley.com/doi/10.1111/bph.v179.12/issuetoc.


Serum hCG-β levels of postovulatory day 12 and 14 with the sequential application of hCG-β fold change significantly increased predictability of pregnancy outcome after IVF-ET cycle.
To investigate hCG-β level on postovulatory day (POD) 12 and its fold increase as predictors for pregnancy outcome after in vitro fertilization (IVF) cycles. A retrospective cohort study was performed in total 1408 fresh and 598 frozen cycles between November 2008 and October 2011, which resulted in biochemical pregnancy, early pregnancy loss, or live birth of singleton pregnancy. The serum hCG-β levels of POD 12 and 14 were compared among biochemical pregnancy, early pregnancy loss, and live birth groups. The cutoff values of POD 12 and 14 hCG-β levels and the degree of hCG-β increase from POD 12 to 14 were determined for each pregnancy outcome. POD 12 and 14 hCG-β levels stratified based on pregnancy outcomes were significantly different among the biochemical pregnancy, early pregnancy loss, and live birth in both fresh and frozen cycles. Serum hCG-β levels of POD 12 and 14 and the fold increase of hCG-β levels from POD 12 to 14 significantly predict pregnancy outcomes after fresh and frozen cycles. Among these, the cutoff value of POD 14 hCG-β had the highest sensitivity and positive predictive value (PPV). In fresh cycles, the cutoff values of POD 12 and 14 serum hCG-β levels for clinical pregnancies were 30.2 mIU/mL (sensitivity 81.3 %, specificity 79.6 %, and PPV 92.3 %) and 70.5 mIU/mL (sensitivity 88.4 %, specificity 85.2 %, and PPV 94.7 %). In pregnancies with POD 12 serum hCG-β levels ≥30.2 mIU/mL, the cutoff level of increase of hCG-β for clinical pregnancy was 2.56 (sensitivity 73.6 %, specificity 72.4 %, and PPV 97.8 %). Sequential application of cutoff values such as POD 12 hCG-β and fold increase of hCG-β improved predictability of pregnancy outcome as compared with that of POD 12 hCG-β alone. The cutoff values of POD 12 and 14 serum hCG-β levels for live birth were 40.5 mIU/mL (sensitivity 75.2 %, specificity 72.6 %, PPV 78.9 %) and 104.5 mIU/mL (sensitivity 80.3 %, specificity 74.1 %, PPV 80.8 %). In the frozen cycles, the cutoff values of POD 12 and 14 serum hCG-β level for clinical pregnancy were 31.5 IU/L (sensitivity 80.4 %, specificity 71.1 % and PPV 90 %) and 43.5 mIU/mL (sensitivity 72.6 %, specificity 71.7 %, PPV 77.2 %). In pregnancies with POD 12 serum hCG-β level ≥31.5 mIU/mL, the cutoff value for fold increase of hCG-β was 2.38 for clinical pregnancy (sensitivity 81.6 %, specificity 71.4 % and PPV 87.9 %). The cutoff values of POD 12 and 14 for live birth were 43.5 mIU/mL (sensitivity 72.6 %, specificity 71.7 %, PPV 77.2 %) and 101.6 mIU/mL (sensitivity 79.6 %, specificity 71.1 %, PPV 78.4 %). Sequential application of cutoff values for POD 12 hCG-β level and fold increase of hCG-β significantly increased PPV for live birth but not clinical pregnancy in frozen cycles. Early prediction of pregnancy outcome by using POD 12 and 14 cutoff levels and sequential application of cutoff value of fold increase could provide appropriate reference to health care providers to initiate earlier management of high-risk pregnancies and precise follow-up of abnormal pregnancies.


A technique for the controlled fracture of micropipette electrode tips.
A method is described for the controlled fracture of electrolyte-filled micropipette electrodes, so that their tip diameter and DC resistance becomes suitable for extracellular recording from individual neurones. The technique is simple and has a low wastage rate. The apparatus used is that employed in conventional microelectrode recording.


The effect of Definity on systemic and pulmonary hemodynamics in patients.
The purpose of this study was to evaluate the pulmonary and systemic hemodynamic effects of Definity in patients with normal as well as those with elevated pulmonary artery pressure at baseline. Secondary objectives of the study were to evaluate safety and determine whether any potential immunologic reactions develop after Definity administration. Patients with normal and elevated pulmonary artery systolic pressure undergoing right-heart catheterization received Definity (10 μL/kg) as a slow bolus over 30 to 60 sec. Multiple sequential measurements of right atrial pressure, pulmonary artery systolic pressure, pulmonary artery diastolic pressure, mean pulmonary artery pressure, cardiac output, and pulmonary capillary wedge pressure were made before and after Definity administration. Vital signs, electrocardiograms, and blood samples were taken at multiple time points. Patients were followed for the development of adverse events. A total of 32 patients (16 with elevated pulmonary artery systolic pressure > 35 mm Hg) were enrolled. No significant changes in any pulmonary or systemic hemodynamic parameters, vital sign values, electrocardiographic data, or laboratory variables were found for data obtained before versus after receipt of Definity. The administration of Definity at the approved dosage does not change pulmonary or systemic hemodynamics in control patients or those with mild to moderate pulmonary hypertension. No significant changes were noted in a wide array of clinical and laboratory safety assessments after patients were exposed to Definity.


A functional account of stimulation-based aerobic glycolysis and its role in interpreting BOLD signal intensity increases in neuroimaging experiments.
In aerobic glycolysis, oxygen is abundant, and yet cells metabolize glucose without using it, decreasing their ATP per glucose yield by 15-fold. During task-based stimulation, aerobic glycolysis occurs in localized brain regions, presenting a puzzle: why produce ATP inefficiently when, all else being equal, evolution should favor the efficient use of metabolic resources? The answer is that all else is not equal. We propose that a tradeoff exists between efficient ATP production and the efficiency with which ATP is spent to transmit information. Aerobic glycolysis, despite yielding little ATP per glucose, may support neuronal signaling in thin (< 0.5 µm), information-efficient axons. We call this the efficiency tradeoff hypothesis. This tradeoff has potential implications for interpretations of task-related BOLD "activation" observed in fMRI. We hypothesize that BOLD "activation" may index local increases in aerobic glycolysis, which support signaling in thin axons carrying "bottom-up" information, or "prediction error"-i.e., the BIAPEM (BOLD increases approximate prediction error metabolism) hypothesis. Finally, we explore implications of our hypotheses for human brain evolution, social behavior, and mental disorders.


Genome wide comparative comprehensive analysis of Plasmodium falciparum MCM family with human host.
Mini chromosome maintenance (MCM) proteins 2-7, a subgroup of the large AAA ATPase family are critically required for eukaryotic DNA replication. These proteins are most likely responsible for unwinding DNA at the replication forks. Besides this function, some MCMs are also involved in other chromosome transactions such as transcription, chromatin remodeling and genome stability. All the MCMs contain a conserved region of ~200 amino acids responsible for nucleotide binding. The importance of MCM proteins is evident by the fact that deregulation of the activity of MCM family of proteins appears to be directly linked to human carcinogenesis. This article will focus on members of this important family of proteins from the malaria parasite Plasmodium falciparum and their comparison with the human host.


Structural studies on the R-type lipopolysaccharide of Aeromonas hydrophila.
A rough strain of Aeromonas hydrophila, AH-901, has an R-type lipopolysaccharide with the complete core. The following core structure was established by chemical degradations followed by sugar and methylation analyses along with ESIMS and NMR spectroscopy: [formula: see text] where D-alpha-D-Hep and l-alpha-D-Hep stand for D-glycero- and l-glycero-alpha-D-manno-heptose, respectively; Kdo stands for 3-deoxy-D-manno-oct-2-ulosonic acid; all monosaccharides are in the pyranose form; the degree of substitution with beta-D-Gal is approximately 50%. Lipid A of the lipopolysaccharide has a 1,4(')-bisphosphorylated beta-D-GlcN-(1-->6)-alpha-D-GlcN disaccharide backbone with both phosphate groups substituted with 4-amino-4-deoxyarabinose residues.


Fluorescent and Bioluminescent Reporter Mouse-Adapted Ebola Viruses Maintain Pathogenicity and Can Be Visualized in Vivo.
Ebola virus (EBOV) causes lethal disease in humans but not in mice. Here, we generated recombinant mouse-adapted (MA) EBOVs, including 1 based on the previously reported serially adapted strain (rMA-EBOV), along with single-reporter rMA-EBOVs expressing either fluorescent (ZsGreen1 [ZsG]) or bioluminescent (nano-luciferase [nLuc]) reporters, and dual-reporter rMA-EBOVs expressing both ZsG and nLuc. No detriment to viral growth in vitro was seen with inclusion of MA-associated mutations or reporter proteins. In CD-1 mice, infection with MA-EBOV, rMA-EBOV, and single-reporter rMA-EBOVs conferred 100% lethality; infection with dual-reporter rMA-EBOV resulted in 73% lethality. Bioluminescent signal from rMA-EBOV expressing nLuc was detected in vivo and ex vivo using the IVIS Spectrum CT. Fluorescent signal from rMA-EBOV expressing ZsG was detected in situ using handheld blue-light transillumination and ex vivo through epi-illumination with the IVIS Spectrum CT. These data support the use of reporter MA-EBOV for studies of Ebola virus in animal disease models.


Application of manganese oxides in wastewater treatment: Biogeochemical Mn cycling driven by bacteria.
Manganese oxides (MnOx) are recognized as a strongest oxidant and adsorbent, of which composites have been proved to be effective in the removal of contaminants from wastewater. This review provides a comprehensive analysis of Mn biochemistry in water environment including Mn oxidation and Mn reduction. The recent research on the application of MnOx in the wastewater treatment was summarized, including the involvement of organic micropollutant degradation, the transformation of nitrogen and phosphorus, the fate of sulfur and the methane mitigation. In addition to the adsorption capacity, the Mn cycling mediated by Mn(II) oxidizing bacteria and Mn(IV) reducing bacteria is the driving force for the MnOx utilization. The common category, characteristics and functions of Mn microorganisms in recent studies were also reviewed. Finally, the discussion on the influence factors, microbial response, reaction mechanism and potential risk of MnOx application in pollutants' transformation were proposed, which might be the promising opportunities for the future investigation of MnOx application in wastewater treatment.


Gene therapy for hair cell regeneration: Review and new data.
Hair cells (HCs) in the cochlea are responsible for transducing mechanical sound energy into neural impulses which lead to the perception of sound. Loss of these sensory cells is the most common cause of sensorineural hearing loss, and spontaneous HC regeneration does not occur in mature mammals. Among the future potential treatment modalities is gene therapy, which is defined as the administration of either DNAs or RNAs as active pharmaceutical ingredients for inducing a clinically-beneficial response. Gene therapy is being envisioned and evaluated as a potential tool for addressing a number of human inner ear disorders. This paper is a hybrid Review and Research Paper, including unpublished data and a review of HC regeneration studies in live animal models. Current gene therapeutic approaches for replacing lost HC populations have been aimed at converting supporting cells surviving within the neuro-epithelium to new HCs by inducing upregulation of bHLH transcription factors such as Atoh1 or reciprocal silencing of Notch signaling with siRNAs, to tip the balance of transcriptional regulation toward a HC fate. Development of one or more of these techniques may yield a path to effective restoration of inner ear form and function. This review also describes other approaches and molecular targets that may prove efficacious and provides perspectives on future clinical challenges and opportunities for gene therapy to become a valuable weapon for the long-anticipated realization of this regenerative treatment.


Antibody response to bacterial antigens covalently bound to biodegradable polymerized serum albumin beads.
Model vaccines have been made by covalently linking Clostridium botulinum type D toxin and Klebsiella pneumoniae capsular polysaccharide antigen to polymerized rabbit serum albumin beads. When injected into rabbits these bead vaccines induced an enhanced production of specific humoral antibody without causing adverse reactions. The adjuvant effect is due to a slow release from the bead structure and offers an alternative to oil emulsions and mineral salt adsorbents.


Insecticide resistance to insect growth regulators, avermectins, spinosyns and diamides in Culex quinquefasciatus in Saudi Arabia.
Culex quinquefasciatus is not only a biting nuisance but also an important vector of fatal diseases. In Saudi Arabia, management measures to control this mosquito and thereby prevent associated disease transmission have focused on insecticides. Nevertheless, information on the resistance status of C. quinquefasciatus is insufficient, especially concerning insecticides containing novel classes of insecticides. We evaluated six insecticides belonging to four classes of insecticides (insect growth regulators [3], avermectins [1], diamides [1] and spinosyns [1]) for toxicity and resistance in eight C. quinquefasciatus populations (from Ishbiliya, Al-Masfa, Al-Masanie, Al-Washlah, Al-Nakhil, Irqah, Al-Suwaidi and Al-Ghanemiya) following World Health Organisation protocols. Resistance status ranging from susceptibility/low resistance to high resistance, in comparison with the susceptible strain, was detected for cyromazine in the eight C. quinquefasciatus populations: Ishbiliya (resistance ratio [RR] = 3.33), Al-Masfa (RR = 4.33), Al-Masanie (RR = 3.67), Al-Washlah (RR = 2.33), Al-Nakhil (RR = 5.33), Irqah (RR = 7.00), Al-Suwaidi (RR = 21.33) and Al-Ghanemiya (RR = 16.00). All C. quinquefasciatus populations exhibited a high level of resistance to diflubenzuron (RR = 13.33-43.33), with the exception of Al-Nakhil which exhibited moderate resistance (RR = 10.00). Susceptibility/low resistance to high resistance was observed for triflumuron in the eight C. quinquefasciatus populations: Ishbiliya (RR = 0.50), Al-Ghanemiya (RR = 3.00), Al-Suwaidi (RR = 10.00), Al-Masfa (RR = 5.00), Al-Masanie (RR = 10.00), Al-Nakhil (RR = 5.00), Irqah (RR = 5.00) and Al-Washlah (RR = 15.00). Susceptibility/low resistance was assessed for abamectin, chlorantraniliprole and spinosad in all C. quinquefasciatus populations, with RR ranges of 0.25-3.50, 0.17-2.19, and 0.02-0.50, respectively. However, the population collected from Irqah showed high resistance to chlorantraniliprole (RR = 11.93). The detection of widespread resistance to insect growth regulators in C. quinquefasciatus highlights an urgent need to establish integrated vector management strategies. Our results may facilitate the selection of potent insecticides for integrated vector management programmes for C. quinquefasciatus.


Reduction of myocardial infarct size in rats under the effect of bepridil.
The ability of intravenous or oral bepridil to reduce infarct size was studied in the rat submitted in situ to coronary-artery ligation. Infarct size was measured by planimetry of serial 8-micron sections at known intervals 48 hr after left-coronary-artery ligation. Succinate dehydrogenase activity (nitroblue tetrazolium stain) was used as an index of tissue viability. Bepridil was administered either intravenously (5 mg/kg) 15 min before (pretreatment) or 10 min after (post-ligation treatment) coronary-artery ligation, or orally (50 mg/kg per day) for 8 days before coronary ligation. Intravenous bepridil pretreatment reduced infarct size by more than 30%. This reduction was accompanied by a better preservation of the myocardial content of adenine nucleotides and a reduction in lactate accumulation. Intravenous post-ligation treatment and chronic oral treatment induced a reduction in infarct size similar to the one observed with intravenous preligation treatment. Thus, bepridil is equipotent in reducing infarct size whether the drug is administered intravenously before or after coronary-artery occlusion, or orally before coronary occlusion.


Targeting nanomedicines in the treatment of rheumatoid arthritis: focus on certolizumab pegol.
Anti-TNFalpha therapy has revolutionized the treatment of rheumatoid arthritis (RA) and other inflammatory diseases. These drugs are powerful and expensive. A new anti-TNFalpha agent, a nanomolecule comprising a humanized Fab' antibody fragment against TNFalpha with a polyethylene glycol tail, is shortly to complete phase III trials in RA. In this review we will discuss the construct of this new molecule, data from trials so far, and its potential place in the market place.


Characterization of projections from a sexually dimorphic motor nucleus in the spinal cord of adult green anoles.
Male green anoles possess two copulatory organs (hemipenes), which are independently controlled by bilateral muscles: the transversus penis (TPN) and retractor penis magnus (RPM). Adult females do not possess hemipenes or either of the two related muscles. Motoneurons projecting to the TPN lie in spinal segments trunk 17 and sacral 1 (T17-S1). Overall, motoneurons in this region are larger and more numerous in males than females. The present studies were designed to determine 1) whether motoneurons projecting to the RPM are located in the same sexually dimorphic nucleus, 2) other targets of T17-S1 motoneurons, and 3) the approximate proportion of motoneurons projecting to each muscle. In Study 1, unilateral injection of the retrograde tracer Fast Blue (FB) into RPMs and simultaneous unilateral injection of either Cholera Toxin-fluorescein (CT-FITC) or Diamidino Yellow into TPNs revealed that RPM and TPN motoneurons are indeed interdigitated in T17-S1. In Study 2, FB was used to characterize other targets of this nucleus in both males and females. In adult males, projections to four muscles accounted for 96% of the T17-S1 motoneurons: the TPN, RPM, caudifemoralis (CF), and cloacal sphincter (SC). In adult females, projections to the CF and SC comprised 70% of this nucleus. These data demonstrate that the T17-S1 nucleus is a mixed spinal nucleus that has projections to muscles present in both sexes, as well as those present only in males and specialized for male copulatory behavior.


Combined cytotoxic and endocrine therapy in postmenopausal patients with advanced breast cancer. A randomized study of CMF vs CMF plus tamoxifen.
In this study 263 patients with advanced measurable breast cancer were randomized to receive cyclophosphamide, methotrexate and 5-fluorouracil (CMF) or CMF + tamoxifen (T). Each cycle of CMF (C, 100 mg/m2 p.o. days 1-14, M 40 mg/m2 i.v. days 1 and 8, F, 600 mg/m2 i.v. days 1 and 8) was repeated every 4 weeks. Tamoxifen, 20 mg twice daily, was given continuously. The treatment results have been assessed by external review for the 220 evaluable patients and were the following for the CMF and CMF + T groups, respectively: PD: 24 and 10%; NC: 27 and 15%; PR: 29 and 44%; and CR: 20 and 31%. The difference between response (CR + PR) rates is highly significant (P = 0.0001). The median duration of remission was 12 months in the CMF-treated group and 18 months in patients treated with CMF + T (P = 0.09). The median duration of survival was 19 months and 24 months respectively (P = 0.07). However, in the group of responders patients receiving CMF + T survived significantly longer than those who received CMF alone (32 months vs 21 months, P = 0.005). The addition of T appeared to be of benefit in all subgroups. The largest differences were seen in patients with the dominant site of disease in the viscera, in patients with a Karnofsky index of 100, and in patients of more than 60 yr of age. The amount of CMF given was identical in the two treatment groups and there was a trend toward a decrease in dose with increasing age. No relationship between response and dose given was observed. In conclusion, the simultaneous use of T and CMF improves the therapeutic results in patients with advanced breast cancer.


Time to endoscopic vacuum therapy-lessons learned after > 150 robotic-assisted minimally invasive esophagectomies (RAMIE) at a German high-volume center.
In esophageal surgery, anastomotic leak (AL) remains one of the most severe and critical adverse events after oncological esophagectomy. Endoscopic vacuum therapy (EVT) can be used to treat AL; however, in the current literature, treatment outcomes and reports on how to use this novel technique are scarce. The aim of this study was to evaluate the outcomes of patients with an AL after IL RAMIE and to determine whether using EVT as an treatment option is safe and feasible. This study includes all patients who developed an Esophagectomy Complications Consensus Group (ECCG) type II AL after IL RAMIE at our center between April 2017 and December 2021. The analysis focuses on time to EVT, duration of EVT, and follow up treatments for these patients. A total of 157 patients underwent an IL RAMIE at our hospital. 21 patients of these (13.4%) developed an ECCG type II AL. One patient died of unrelated Covid-19 pneumonia and was excluded from the study cohort. The mean duration of EVT was 12 days (range 4-28 days), with a mean of two sponge changes (range 0-5 changes). AL was diagnosed at a mean of 8 days post-surgery (range 2-16 days). Closure of the AL with EVT was successful in 15 out of 20 patients (75%). Placement of a SEMS (Self-expandlable metallic stent) after EVT was performed in four patients due to persisting AL. Overall success rate of anastomotic sealing independently of the treatment modality was achieved in 19 out of 20 Patients (95%). No severe EVT-related adverse events occurred. This study shows that EVT can be a safe and effective endoscopic treatment option for ECCG type II AL.


Positive effects of refugee presence on host community nutritional status in Turkana County, Kenya.
Refugee camps are often assumed to negatively impact local host communities through resource competition and conflict. We ask instead whether economic resources and trade networks associated with refugees have benefits for host community health and nutrition. To address this question we assess the impacts of Kakuma Refugee Camp in northwest Kenya, comparing anthropometric indicators of nutritional status between Turkana communities in the region. Participants were recruited at four sites in Turkana County (N = 586): Kakuma Town, adjacent to Kakuma Refugee Camp; Lorugum, an area with sustained economic development; Lokichoggio, formerly host to international NGOs, and now underdeveloped; and Lorengo, an undeveloped, rural community. We evaluated nutritional status using summed skinfold thickness and body mass index (BMI). Structured interviews provided contextual data. Age-controlled multiple regression models reveal two distinct skinfold thickness profiles for both sexes: comparatively elevated values in Kakuma and Lorugum, and significantly lower values in Lorengo and Lokichoggio. BMI did not vary significantly by location. Despite better nutritional status, a large proportion of Kakuma residents still report worries about basic needs, including hunger, health, and economic security. Kakuma Refugee Camp is associated with better host community energetic status indicators, compared to other relevant, regional sites varying in development and resources. Based on global nutritional standards, observed differences likely represent meaningful disparities in overall health. We suggest that access to cereals via refugee trade networks and employment might mediate this relationship. However, perceptions of refugees as illegitimate interlopers maintain a high psychological burden.


A bioinformatic analysis of WFDC2 (HE4) expression in high grade serous ovarian cancer reveals tumor-specific changes in metabolic and extracellular matrix gene expression.
Human epididymis protein-4 (HE4/WFDC2) has been well-studied as an ovarian cancer clinical biomarker. To improve our understanding of its functional role in high grade serous ovarian cancer, we determined transcriptomic differences between ovarian tumors with high- versus low-WFDC2 mRNA levels in The Cancer Genome Atlas dataset. High-WFDC2 transcript levels were significantly associated with reduced survival in stage III/IV serous ovarian cancer patients. Differential expression and correlation analyses revealed secretory leukocyte peptidase inhibitor (SLPI/WFDC4) as the gene most positively correlated with WFDC2, while A kinase anchor protein-12 was most negatively correlated. WFDC2 and SLPI were strongly correlated across many cancers. Gene ontology analysis revealed enrichment of oxidative phosphorylation in differentially expressed genes associated with high-WFDC2 levels, while extracellular matrix organization was enriched among genes associated with low-WFDC2 levels. Immune cell subsets found to be positively correlated with WFDC2 levels were B cells and plasmacytoid dendritic cells, while neutrophils and endothelial cells were negatively correlated with WFDC2. Results were compared with DepMap cell culture gene expression data. Gene ontology analysis of k-means clustering revealed that genes associated with low-WFDC2 were also enriched in extracellular matrix and adhesion categories, while high-WFDC2 genes were enriched in epithelial cell proliferation and peptidase activity. These results support previous findings regarding the effect of HE4/WFDC2 on ovarian cancer pathogenesis in cell lines and mouse models, while adding another layer of complexity to its potential functions in ovarian tumor tissue. Further experimental explorations of these findings in the context of the tumor microenvironment are merited.


Medical Students' Experiences and Perspectives on Interpreting for LEP Patients at Two US Medical Schools.
Language concordance between patient and provider has been shown to improve health outcomes for Limited English Proficiency (LEP) patients. However, health care teams often use available ad hoc interpreters without knowing whether their language skills are adequate. Little is known about the role of medical students working as ad hoc interpreters. Bilingual medical students are engaged as interpreters in the care of LEP patients and may serve as a potential resource for health care teams caring for LEP patients. We conducted a multi-institutional online survey of graduating medical students at the University of Illinois and the University of Chicago in 2011 and 2012, containing both qualitative and quantitative questions regarding their experiences as interpreters for LEP patients. Half (216/430) of contacted students completed the survey; 40 % (87/216) of responding students reported being bilingual. Of these students, the vast majority, 84 % (73/87), had been asked to interpret for patients in the clinical setting. Only 12 % (10/87) of students reported having felt uncomfortable interpreting for patients "often" or "very often." Over half (53 %, 46/87) described incidents during which they felt uncomfortable interpreting. Seventeen (17/46, 37 %) students described those incidents as high-stakes clinical settings. Medical schools and health care institutions should establish guidelines for students who identify as fluent in another language and are interested in interpreting for LEP patients in clinical settings, to protect both students and patients when language poses a barrier to quality care.


Characteristics and Trends of Highly Cited Articles in Calcaneus Fracture Research.
Despite significant effort expended toward exploring fracture patterns, surgical timing, surgical approaches, and possible implants within the calcaneus fracture literature, treatment is still fraught with complications and controversy. This study aims to conduct a citation analysis of the most cited articles related to calcaneus fractures to highlight the most historically influential articles, as well as the more recent breakthrough articles that are leading change within the field. A literature search was performed via Scopus on September 20, 2021 using the terms "(calcaneus OR calcaneal OR hindfoot) AND (fracture OR injury)" to search "article title, abstract, and keywords" of all primary and review articles. Search results were rigorously reviewed to ensure appropriateness for this study. The 50 highest total cited included articles were analyzed. Title, authors, journal, country of origin, institution, year of publication, citation variables (total citation count, total citation density, citation count from the last 5 years, and 5-year citation density), and level of evidence were collected for each article. One-way analysis of variance with post hoc testing was used to identify differences in citation variables and level of evidence. Pearson correlation was used to directly compare different citation variables. The total citation count average was 178±33, with a total citation density average of 9±2. The year of publication for the 50 articles ranged from 1948 to 2014, with a median year of 1999. Roy W. Sanders was the most productive author in the field, authoring 6 articles and lead authoring 3. The most frequent level of evidence was IV with 19 articles. There was a strong correlation between total citation density and 5-year citation density. The article level of evidence showed no impact on the included measures of an article's influence. This study successfully analyzes and presents the characteristics among the highest cited articles related to calcaneus fractures. The provided characterization of influential works and authors highlights trends, impactful findings, and future areas of focus within calcaneus fracture literature.Level of Evidence: Review Article.


Allosteric potentiation of GABAA receptor single-channel conductance by netrin-1 during neuronal-excitation-induced inhibitory synaptic homeostasis.
As the principal receptor that mediates both synaptic and tonic inhibition of neurons in the brain, the A-type gamma-aminobutyric acid receptor (GABAAR) is functionally important for maintaining the balance between neuronal excitation and inhibition. Here, we report the identification of netrin-1 as an endogenous allosteric modulator of GABAARs. Following increased neuronal excitability, netrin-1 is secreted and binds to the extracellular domains of GABAAR subunits, thereby inducing homeostatic upscaling of GABAAR-mediated synaptic efficacy and currents. Surprisingly, this homeostatic plasticity is primarily mediated by increasing GABAAR single-channel conductance. Our study reveals an important role of netrin-1 as an endogenous GABAAR allosteric modulator in maintaining neuronal excitation-inhibition balance, a fundamental process for brain function and dysfunction.


Usefulness of Stress Echocardiography in the Management of Patients Treated with Anticancer Drugs.
In recent years, the survival of patients with cancer has improved thanks to advances in antineoplastic therapeutic protocols. This has led to an increasing burden of cardiovascular complications related to cancer treatment. Therefore, a new branch of cardiology has been created, "cardio-oncology," with the aims of preventing cardiovascular complications related to antineoplastic treatment, achieving early diagnosis and treatment of any complications, and allowing completion of the expected antineoplastic treatment. Stress echocardiography has a pivotal role in achieving a timely diagnosis of coronary artery disease and thus is the best management approach in this clinical setting. Atherosclerotic processes can be exacerbated by both chemotherapy and chest irradiation in patients with cancer, even several years after anticancer treatment completion. Moreover, stress echocardiography has many other potential applications, such as in the evaluation of subclinical left ventricular dysfunction and contractile reserve in patients treated with anticancer drugs that have the potential to induce myocardial damage, as well as evaluating valve disease. The objective of this review is to delineate the role of stress echocardiography in cardio-oncology.


Dexamethasone reduces cell surface levels of CD11b on human eosinophils.
Overnight incubation of human eosinophils (Eøs) with the glucocorticoid hormone dexamethasone (DEX; 0.1 microM) resulted in lower expression of the CD11b, but not CD49d, antigen on their plasma membrane, as assessed by flow cytometry. DEX produced a consistent inhibitory effect (ranging from 16% to 20%) when tested at a concentration of 0.1 microM. Eø stimulation with 100 ng/ml eotaxin produced an increase in CD11b (+26%), but not CD11c, levels and concomitantly a reduction (-25%) on CD62L expression. The inhibition exerted by DEX upon CD11b levels was also evident following eotaxin upregulation, with a degree of inhibition similar to that seen on basal levels. These data highlight a novel mechanism of action by which glucocorticoid hormones may be effective in reducing Eø accumulation during allergic inflammation in man.


The use of a monoclonal i-j-specific antibody to distinguish cells in the feedback suppression circuit from those in the contrasuppressor circuit.
A monoclonal anti-I-Jb serum designated D-7 reacts in high titer with three different T-cell subsets and one cell-free product involved in generating contrasuppressive activity, but has no activity against I-J T-cell subsets (or their cell-free mediators) involved in feedback suppression. These results give evidence for heterogeneity in the I-J subregion. They also indicate that the serological markers on I-J+ cells may define the functional activity of the regulatory circuits they belong to. Clearly, they do not separate the role that the cells play within a particular immunoregulatory circuit, i.e., inducer, transducer, or effector cells.


Transcription factor networks in trophoblast development.
The placenta sustains embryonic development and is critical for a successful pregnancy outcome. It provides the site of exchange between the mother and the embryo, has immunological functions and is a vital endocrine organ. To perform these diverse roles, the placenta comprises highly specialized trophoblast cell types, including syncytiotrophoblast and extravillous trophoblast. The coordinated actions of transcription factors (TFs) regulate their emergence during development, subsequent specialization, and identity. These TFs integrate diverse signaling cues, form TF networks, associate with chromatin remodeling and modifying factors, and collectively determine the cell type-specific characteristics. Here, we summarize the general properties of TFs, provide an overview of TFs involved in the development and function of the human trophoblast, and address similarities and differences to their murine orthologs. In addition, we discuss how the recent establishment of human in vitro models combined with -omics approaches propel our knowledge and transform the human trophoblast field.


The quality of nursing in intensive care: a development of a rating scale.
The evaluation of nursing care is a topic of great interest and especially crucial in intensive care contexts. However, inside the Italian scientific scenario it is still difficult to measure NSO, or Nursing Sensitive Outcomes, due to the lack of indicators or scales shared by the nursing community. The aim of the present study was therefore to develop a Quality Nursing Care Scale for the Intensive Care (ICU-I-QNCS). From the literature review of the Intensive Care Unit (ICU) quality standards, they were generated 63 items. Then 43 experts assessed them through the Content Validity Index (CVI). Items with a CVI score <0.90 were removed from the scale. All the 63 items have achieved an average score CVI equal or greater than 0.90. 5 item reached an optimal average CVI score (=1); 23 showed an average CVI score between 0.90-0.94 and last 35 were between 0.95-0.99. The ICU-I-QNCS has obtained an acceptable CVI level and it reflects the underlying theoretical model of Doran (2002).


Reproductive characteristics of the Yangtze vole (Microtus fortis calamorum) under laboratory feeding conditions.
The reproductive characteristics of a laboratory population of the vole Microtus fortis calamorum were examined. Voles were allowed to breed under laboratory feeding conditions. Over a period of 3 months, 61.82% of the 110 vole pairs examined produced 3 or 4 litters. There were 1-9 voles in each litter and the mean litter size was 4.67±0.28 (mean±SE). Most litters included 3-7 young voles, accounting for 83.62% of all litters. The mean farrowing interval was 25.9 days (range from 19 to 95 days), and the most farrowing intervals were 20-25 days, accounting for 79.9% of the total. When based on litter size, the reproductive index was 6.23, but was 3.42 when based on pup survival. The survival rate of offspring to weaning was 55.03%. The high rate of infanticide that occurred after removal of males from cages indicates that, in the laboratory, both parents need to be present prior to weaning.


Does mianserin induce involuntary movements in brain damaged patients?
A case of involuntary movements reappearing during mianserin treatment after 33 years of absence in a 69-year-old woman is described. A noradrenergic alpha-2-receptor-mediated mechanism is suggested.


Tinea capitis at the University Hospital of Tizi-Ouzou, Algeria, and first isolation of Trichophyton tonsurans.
Tinea capitis, which are fungal infections caused by some dermatophyte species, are common in developing countries, such as Algeria, where they represent a public health concern. In order to define the epidemiological, clinical and diagnostic features of these infections, a prospective study was conducting from September 2018 to May 2019, at the University Hospital of Tizi-Ouzou (Algeria). All patients addressed to the Laboratory of Parasitology-Mycology of the University Hospital of Tizi-Ouzou for a suspected Tinea capitis, were included in this study. Before sampling, contact with animals or soil, presence of similar lesions in the family circle, and previous antifungal or corticosteroid treatment were searched. Mycological examination included direct microscopic examination of the samples, and culture on Sabouraud agar slants at 27°C for up to 4weeks. Out of the 87 samples examined, 46 allowed us to confirm the diagnosis of Tinea capitis, representing a positivity rate of 52.9%. The sex ratio was 1.09 (52.2% males vs. 47.8% females among the infected patients), and the infection mainly involved children of 4-6years (43.3%). Thirty-four strains of dermatophytes were isolated. Microsporum canis was the most frequent species identified (44.1%), followed by Trichophyton mentagrophytes (38.2%). During this study, Trichophyton tonsurans, an unusual dermatophyte species in Algeria, was identified for the first time in our hospital. Tinea capitis are still common in Algeria, mainly affecting school-aged children and preschool children. Microsporum canis and T. mentagrophytes are the major causative agents, in agreement with previous studies showing a decrease in frequency of anthropophilic species, in favour of zoophilic species.


Biomechanical evaluation of abutment stability in morse taper implant connections in different times: A retrospective clinical study compared with an in vitro analysis.
Micromotion between a dental implant and abutment can adversely affect clinical performance and compromise successful osseointegration by creating a bacterial harbor, enabling screw loosening, and imparting disruptive lateral forces on the cortical bone. Thus, the aim of the present study was to measure the abutment stability evolution using resonance frequency analysis (RFA) in vivo at four different times (baseline, 3, 4, and 12 months), and compare these data obtained with the RFA measured after mechanical cycling (in vitro) corresponding to the proposed times in numbers of cycles. To evaluate the abutment stability, RFA was performed in 70 sets of implant/abutment (IA) with a total of 54 patients (31 women, 23 men). These IA sets were divided into three groups, according to the abutment angulation: straight abutment (Abt1 group), 17-degree angled abutment (Abt2 group), and 30-degree angled abutment (Abt3 group). Abutment stability was measured immediately at implant placement and the abutment installation (T1), 3 (T2), 4 (T3), and 12 months (T4) later. For the in vitro analysis, ten sets of each group were submitted to mechanical cycling: T1 = 0 cycles, T2 = 90,000 cycles, T3 = 120,000 cycles, and T4 = 360,000 cycles. All data collected were statistically evaluated using the GraphPad Prism 5.01 software, with the level of significance was α = 0.05. In vivo, the overall data of implant stability quotient (ISQ) values obtained for all groups in each evaluation time were 61.5 ± 3.94 (95% CI: [60-63]) at T1, 62.8 ± 3.73 (95% CI, [61-64]) at T2, 63.4 ± 3.08 (95% CI: [61-64]) at T3, and 65.5 ± 4.33 (95% CI: [63-68]) at T4. Whereas in vitro, the ISQ were 61.5 ± 2.66 (95% CI: [59-63]) at T1, 63.2 ± 3.02 (95% CI, [61-65]) at T2, 63.9 ± 2.55 (95% CI: [62-66]) at T3, and 66.5 ± 2.97 (95% CI: [64-68]) at T4. In both evaluations (in vivo and in vitro), the data showed a significant difference (ANOVA test with p < 0.0001). The RFA to measure the abutment stability used in this study showed that there was a progressive increase in stability among the predetermined times for the measurements, in both analysis (in vivo and in vitro). Furthermore, the values at each time point were similar, with no statistical difference between them.


Can transanal natural orifice specimen extraction after laparoscopic anterior resection for colorectal cancer reduce the inflammatory response?
Avoiding abdominal incisions is one of the unique features of natural orifice specimen extraction (NOSE) surgery. There has been no consensus on whether the avoidance of abdominal incisions can reduce the systemic inflammatory response after NOSE surgery. This study was designed to evaluate the safety and feasibility of transanal NOSE, especially the inflammatory response after transanal NOSE versus mini-laparotomy (LAP). A total of 172 colorectal cancer patients who underwent transanal NOSE were matched with 172 patients who underwent LAP for colorectal cancer. Clinical characteristics, pathological features, perioperative parameters, and indicators of the inflammatory response were collected and compared. The inflammatory response was assessed by measuring body temperature, neutrophil count, C-reactive protein levels, and procalcitonin levels. Patients in the NOSE group had better short-term outcomes, such as lower incidence of wound infection, less postoperative pain, less need for anesthetic drugs, and faster recovery of intestinal function. Regarding the inflammatory response, the average body temperature of patients in the NOSE group was higher on postoperative day (POD) 2 than that in the LAP group. A higher median neutrophil count and C-reactive protein levels were observed in the NOSE group on POD3 and POD5 than was observed in the LAP group. Transanal NOSE is safe and feasible for colorectal cancer, with better short-term outcomes. Although transanal NOSE produced a larger systemic inflammatory response than LAP in the early postoperative stages, this response did not appear to translate into infectious morbidity.


Evidence-based Urology: Understanding Heterogeneity in Systematic Reviews.
Variability in the results of randomized trials presents challenges to the interpretation and application of the evidence to patient care. Understanding how systematic reviews deal with this problem of "heterogeneity" will help clinicians in applying results in their patient management. This manuscript offers a review of heterogeneity from the clinical urological perspective. PATIENT SUMMARY: Systematic reviews of the literature are necessary to accurately summarize the available evidence to inform clinical decisions. In this mini-review, we explain how to understand and deal with the differences between studies-which we call heterogeneity-included in these types of reviews.


Flow analysis of heavy metals in a pilot-scale incinerator for residues from waste electrical and electronic equipment dismantling.
The large amount of residues generated from dismantling waste electrical and electronic equipment (WEEE) results in a considerable environmental burden. We used material flow analysis to investigate heavy metal behavior in an incineration plant in China used exclusively to incinerate residues from WEEE dismantling. The heavy metals tested were enriched in the bottom and fly ashes after incineration. However, the contents of heavy metals in the bottom ash, fly ash and exhaust gas do not have a significant correlation with that of the input waste. The evaporation and recondensation behavior of heavy metals caused their contents to differ with air pollution control equipment because of the temperature difference during gas venting. Among the heavy metals tested, Cd had the strongest tendency to transfer during incineration (TCd=69.5%) because it had the lowest melting point, followed by Cu, Ni, Pb and Zn. The exchangeable and residual fractions of heavy metals increased substantially in the incineration products compared with that of the input residues. Although the mass of residues from WEEE dismantling can be reduced by 70% by incineration, the safe disposal of the metal-enriched bottom and fly ashes is still required.


Two-dimensional Turbulence in Symmetric Binary-Fluid Mixtures: Coarsening Arrest by the Inverse Cascade.
We study two-dimensional (2D) binary-fluid turbulence by carrying out an extensive direct numerical simulation (DNS) of the forced, statistically steady turbulence in the coupled Cahn-Hilliard and Navier-Stokes equations. In the absence of any coupling, we choose parameters that lead (a) to spinodal decomposition and domain growth, which is characterized by the spatiotemporal evolution of the Cahn-Hilliard order parameter ϕ, and (b) the formation of an inverse-energy-cascade regime in the energy spectrum E(k), in which energy cascades towards wave numbers k that are smaller than the energy-injection scale kin j in the turbulent fluid. We show that the Cahn-Hilliard-Navier-Stokes coupling leads to an arrest of phase separation at a length scale Lc, which we evaluate from S(k), the spectrum of the fluctuations of ϕ. We demonstrate that (a) Lc ~ LH, the Hinze scale that follows from balancing inertial and interfacial-tension forces, and (b) Lc is independent, within error bars, of the diffusivity D. We elucidate how this coupling modifies E(k) by blocking the inverse energy cascade at a wavenumber kc, which we show is ≃2π/Lc. We compare our work with earlier studies of this problem.


High efficiency of ferricenium salts as radiosensitizers of V79 cells in vitro and the KHT tumor in vivo.
Ferricenium salts (Fc+X-) can act as radiosensitizers of hypoxic cells in vitro and of KHT tumor cells in vivo. Sensitization is manifest by a removal of the shoulder of the hypoxic radiation survival curve in vitro. This results in a dose modification factor (DMF) of 2.0 when measured at a surviving fraction of 0.1, for 10 mumol dm-3 FcPF6. In comparison, little sensitization is seen in air. The toxicity of these metal complexes, but not their radiosensitizing ability, can be decreased by including protein (Bovine serum albumin) in the medium. This may be related to the fact that the ferricenium complexes react readily with nucleophiles to generate OH radicals. Ferricenium trichloracetate, when given at a dose of 200 mg/kg to mice with the KHT tumor gives an enhancement ratio of 1.3.


Comparison of murine lymphokine-activated killer cells, natural killer cells, and cytotoxic T lymphocytes.
Precursors and effectors of murine lymphokine-activated killer cells, natural killer cells, and cytotoxic T lymphocytes are compared. Natural killer cells are resistant to gamma-irradiation (1000 R) whereas precursors of lymphokine-activated killer cells and cytotoxic T lymphocytes are sensitive. Lower doses of gamma-irradiation (500 R) remove precursors for cytotoxic T lymphocytes but not lymphokine-activated killer cells. In addition, lymphokine-activated killer cells are regenerated before classical CTL after sublethal doses of gamma-irradiation. Natural killer cells are resistant to anti-Thy 1 and C' and anti-thymocyte serum, but sensitive to anti-asialo GM1 and complement. Precursors of cytotoxic T lymphocytes are sensitive to anti-Thy 1 and complement and anti-thymocyte serum, but are resistant to anti-asialo GM1 and complement. Precursors of lymphokine-activated killer cells are partially sensitive to anti-Thy 1 and complement and anti-thymocyte serum, but are resistant to anti-asialo GM1 and complement. Effector cells of cytotoxic T lymphocytes are sensitive to anti-Thy 1 and complement and resistant to anti-asialo GM1 and complement. Lymphokine-activated killer cell effectors are sensitive to anti-asialo GM1 and complement at 24 hr after activation. These effectors are more closely aligned with classical natural killer effectors. Lymphokine-activated killer effectors, 7 days after activation, are resistant to anti-asialo GM1 and complement and sensitive to anti-Thy 1 and complement. Relationships and differences among these cytotoxic subsets are discussed.


Tyrosine Kinase Inhibitor discontinuation in Chronic Myeloid Leukemia: eligibility criteria and predictors of success.
TKI discontinuation proved to be safe and feasible in patients with CML with deep and durable molecular responses, introducing an additional treatment goal for these patients beyond overall survival. However, treatment interruption is a safe procedure only with appropriate patient selection and monitoring. Clinical and biological factors associated with better outcomes do not yet offer a precise stratification of patients according to their risk of relapse. This article aims at reviewing the leading studies present in the field in order to define eligibility criteria for discontinuation and predictors of success.


Multilevel instrumented posterolateral lumbar spine fusion with an allogeneic cellular bone graft.
Low back pain (LBP) is the leading cause of absence from work, disability, and impaired quality of life. Fusion surgery may be indicated when non-operative treatments have failed to provide relief. Surgery may include the use of fusion-enhancing implants, such as cellular bone allografts (CBAs). The purpose of this retrospective study was to evaluate efficacy and safety of one CBA (V-CBA) in patients who underwent instrumented posterolateral fusion (IPLF). Retrospective data were collected from 150 consecutive patients who had undergone IPLF surgery between January 1, 2015, and March 31, 2018, in which V-CBA was used. All surgeries were performed by one surgeon. V-CBA was mixed with local autograft bone. Patient diagnoses included degenerative disc disease, spondylosis, spondylolisthesis, or spondylolysis with or without stenosis. Standing anteroposterior (AP) and lateral images were collected prior to surgery and again at the terminal visit, which took place between 6 and 33 months post-operatively. De-identified images were assessed radiologically. Adverse events were documented. The primary composite endpoint of fusion status was dependent upon two main criteria: bridging bone per the Lenke scale (classified as "A" definitely solid or "B" possibly solid) and posterior hardware status (intact). Lenke scale C or D were categorized as pseudarthrosis. Eighty-seven male and 63 female patients (613 levels total) underwent IPLF in which V-CBA was implanted. An average of 4.1 levels was treated, with 59.3% of patients having undergone treatment for more than 3 levels. Twenty-nine percent of patients had diabetes. Fifty-two percent of patients had previously used nicotine products, and 12% were current smokers. Sixteen serious adverse events were recorded and included lumbar seroma, cerebrospinal fluid leak, wound dehiscence, pneumonia, urinary tract infection, and myocardial infarction. Successful fusion (Lenke scale "A" or "B") was recorded in 148 out of 150 patients (98.7%), or 608 out of 613 levels. The total pseudarthrosis rate was 0.8%. The use of V-CBA combined with local autograft in multilevel IPLF resulted in successful fusions in 98.7% of patients. These results are particularly robust given the complex nature of many of these cases: 89 patients had 4 or more surgical levels, and many patients had multiple comorbidities. IV.


Apparent adoptive immune enhancement by bone marrow and bursa of Fabricius cells from juvenile chickens.
The occurrence of enhancer cells (EC) in the bone marrow and bursa of Fabricius of unprimed and mouse erythrocyte (MRBC) primed birds was investigated. EC are defined operationally as immunocompetent cells that are incapable of adoptive immunity in embryo hosts but able to enhance the immune responsiveness of newly-hatched chick recipients. Weak to moderate immune enhancement was observed in chick hosts grafted with bursal cells from unprimed or MRBC-primed donors whereas with bone marrow cell transfer the immune enhancement was weak with cells from unprimed donors but modest to strong with those from primed donors. Thus antigen priming of donors had little effect on the EC level of the bursa but appeared to increase that in the bone marrow. Moreover, the EC activity of bone marrow of primed donors was donor-age dependent. Response profile studies revealed that the EC level of donor bone marrow was low the first 2-3 days after immunization and high by day six. The elevated EC level in the bone marrow of immunized donors is believed due to both recruitment of immigrant cells of extramedullary origin and clonal expansion of medullary immunocompetent cells.


Arginine vasopressin remolds the spontaneous discharges disturbed by amyloid β protein in hippocampal CA1 region of rats.
Beta-amyloid peptide (Aβ) aggregated in the brain is the main pathological characteristic of Alzheimer's disease (AD), and a significant decrease in the concentration of arginine vasopressin (AVP) in the brain of AD patients has been reported. Our recent study shows that intracerebroventricular (i.c.v.) injection of AVP protects against Aβ-induced impairments of spatial learning and memory. However, it is still unclear whether the Aβ-induced cognitive deficit is involved in the alteration of central neuronal discharges, and further whether AVP can modulate the electrophysiological change induced by Aβ. The present study thus observed the effects of AVP, Aβ and AVP plus Aβ on the spontaneous discharges of hippocampal CA1 neurons in rats by using multi-channel extracellular recording technique. The results showed that: (1) the average frequency of spontaneous discharges was decreased by i.c.v. injection of 25 nmol Aβ(25-35); (2) 10 nmol AVP induced an increase in spike discharge in the hippocampal CA1 neurons; (3) pretreatment with 10 nmol AVP effectively reversed Aβ(25-35) induced suppression of spontaneous discharges in hippocampal CA1 region. These in vivo electrophysiological results indicate that AVP, as a hormone and neurotransmitter, can remold the spontaneous discharges disturbed by Aβ and counteract the deleterious effect of Aβ(25-35) on neural circuit, suggesting that the activation of central vasopressinergic system may play a beneficial role for the prevention and treatment of cognitive impairments in AD.


Spatiotemporal variation of bacterial assemblages in a shallow subtropical coastal lagoon in Southern Brazil.
A study on the bacterioplankton of Conceição Lagoon (27 degrees 34' S-48 degrees 27' W), Southern Brazil, was carried out in July 2005 (austral winter) and January 2006 (austral summer) to characterize the bacterial spatiotemporal distribution and to determine the heterotrophic and photoautotrophic bacterial dominance in hypoxic/oxic stratified waters. Bacterial abundance increased significantly (p < 0.05) in summer with averages of coccus cyanobacteria (CCY) ranging from 1.02 x 10(5) (winter) to 3.21 x 10(6) cells mL(-1) (summer), heterotrophic coccus/rod-shaped (HCR) cells from 7.00 x 10(4) to 3.60 x 10(6) cells mL(-1), and heterotrophic filamentous (HF) bacteria from 2.90 x 10(3) to 2.74 x 10(5) cells mL(-1). Bacterial biovolumes also increased in summer with mean biovolumes of CCY ranging from 0.38 to 1.37 microm3, HCR cells from 0.31 to 1.12 microm3, and HF from 3.32 to 11.34 microm3. Principal component analysis showed that salinity, temperature, and light were the abiotic factors that better explained the temporal variability of bacterial assemblages. Bacterial heterotrophy dominated in the lagoon, excepted by the southern and part of central sector in January 2006, when autotrophic-dominated microbial community occurred. Spatially, bacterial assemblages were influenced by nutrient gradient, oxygen, and salinity with a positive relationship between biovolumes and nutrients and a negative relationship between abundance of coccus cyanobacteria and nutrients. The stratified [corrected] area revealed a singular temporal pattern with hypoxic bottom waters in winter and oxygen-rich waters appearing in summer related with the availability of light and predominant microbes. Thus, oxygen consumption/production is likely to be regulated by the amount of light reaching the bottom, stimulating the production of oxygen by oxygenic phototrophs.


Assessment of Cell Viability in Electrically Excitable Muscle Cells Through Intact Twitch Stimulation.
Muscle cells (i.e. skeletal muscle fibers) are fully viable and functional when their excitation-contraction (EC) coupling machinery is intact. This involves intact membrane integrity with polarized membrane, functional ion channels for action potential generation and conduction, an intact electro-chemical interface at the level of the fiber's triad, followed by sarcoplasmic reticulum Ca2+ release, and subsequent activation of the chemico-mechanical interface at the level of the contractile apparatus. The ultimate end result is then a visible twitch contraction upon a brief electrical pulse stimulation. For many biomedical studies involving single muscle cells, intact and viable myofibers are of utmost importance. Thus, a simple global screening method that involves a brief electrical stimulus applied to single muscle fibers and assessment of visible contraction would be of high value. In this chapter, we describe step-by-step protocols to (i) obtain intact single muscle fibers from freshly dissected muscle tissue using an enzymatic digestion procedure and (ii) provide a workflow for the assessment of twitch response of single fibers that can be ultimately classified as viable. For this, we have prepared a unique stimulation pen for which we provide the fabrication guide for do-it-yourself rapid prototyping to eliminate the need for expensive specialized commercial equipment.


Target sequence capture data shed light on the deeper evolutionary relationships of subgenus Chamaecerasus in Lonicera (Caprifoliaceae).
The genus Lonicera L. is widely distributed in the north temperate zone and is well-known for its high species richness and morphological diversity. Previous studies have suggested that many sections of Lonicera are not monophyletic and phylogenetic relationships within the genus are still poorly resolved. In this study, we sampled 37 accessions of Lonicera, covering four sections of subgenus Chamaecerasus plus six outgroup taxa, to recover the main clades of Lonicera based on sequences of nuclear loci generated by target enrichment and cpDNA from genome skimming. We found extensive cytonuclear discordance across the subgenus. Both nuclear and plastid phylogenetic analyses supported subgenus Chamaecerasus sister to subgenus Lonicera. Within subgenus Chamaecerasus, sections Isika and Nintooa were each polyphyletic. Based on the nuclear and chloroplast phylogenies, we propose to merge Lonicera korolkowii into section Coeloxylosteum and Lonicera caerulea into section Nintooa. In addition, Lonicera is estimated to have originated in the mid Oligocene (26.45 Ma). The stem age of section Nintooa was estimated to be 17.09 Ma (95% HPD: 13.30-24.45). The stem age of subgenus Lonicera was estimated to be 16.35 Ma (95% HPD: 14.12-23.66). Ancestral area reconstruction analyses indicate that subgenus Chamaecerasus originated in East Asia and Central Asia. In addition, sections Coeloxylosteum and Nintooa originated in East Asia, with subsequent dispersals into other areas. The aridification of the Asian interior likely promoted the rapid radiation of sections Coeloxylosteum and Nintooa within this region. Moreover, our biogeographic analysis fully supports the Bering and the North Atlantic Land Bridge hypotheses for the intercontinental migrations in the Northern Hemisphere. Overall, this study provides new insights into the taxonomically complex lineages of subgenus Chamaecerasus and the process of speciation.


Epigenetic control of heavy metal stress response in mycorrhizal versus non-mycorrhizal poplar plants.
It was previously shown that arbuscular mycorrhizal fungi (AMF) exert a significant improvement of growth in a tolerant white poplar (Populus alba L.) clone (AL35) grown on Cu- and Zn-polluted soil via foliar alterations in the levels of defence/stress-related transcripts and molecules. However, nothing is known about the epigenetic changes which occur during tolerance acquisition in response to heavy metals (HMs) in the same mycorrhizal vs. non-mycorrhizal poplar plants. In order to analyse the epigenome in leaves of AL35 plants inoculated or not with AMF and grown in a greenhouse on multimetal polluted or unpolluted soil, the Methylation Sensitive Amplification Polymorphism (MSAP) approach was adopted to detect cytosine DNA methylation. Modest changes in cytosine methylation patterns were detected at first sampling (4 months from planting), whereas extensive alterations (hypomethylation) occurred at second sampling (after 6 months) in mycorrhizal plants grown in the presence of HMs. The sequencing of MSAP fragments led to the identification of genes belonging to several Gene Ontology categories. Seven MSAP fragments, selected on the basis of DNA methylation status in treated vs control AL35 leaves at the end of the experiment, were analysed for their transcript levels by means of qRT-PCR. Gene expression varied in treated samples relative to controls in response to HMs and/or AMF inoculation; in particular, transcripts of genes involved in RNA processing, cell wall and amino acid metabolism were upregulated in the presence of AMF with or without HMs.


Intestinal T-cell responses to high-molecular-weight glutenins in celiac disease.
The chronic, small intestinal inflammation that defines celiac disease is initiated by a HLA-DQ2 restricted T-cell response to ingested gluten peptides after their in vivo deamidation by tissue transglutaminase (TG2). To date, celiac disease can only be treated by a lifelong abstinence from foods that contain wheat, rye, or barley; better therapeutic options are hence needed. An attractive target would be to identify nontoxic wheat cultivars or components thereof with intact baking qualities. Because these qualities are mainly determined by the high molecular weight (HMW) glutenin proteins of gluten, it is critical to know if these proteins are toxic or, more specifically, if they will trigger the activation of T cells in the celiac lesion. Different, highly purified HMW glutenins were isolated from wheat cultivars or expressed as recombinant proteins. The proteins were first tested for recognition by a large panel of gluten-specific T-cell lines established from celiac lesions and then applied during ex vivo challenges of celiac biopsies to allow for a direct identification of HMW specific T cells. Intestinal T-cell responses to TG2-deamidated HMW glutenins but not the corresponding native proteins were detectable in 9 of the 22 adult and childhood celiac disease patients tested. T cells within celiac lesions frequently recognize deamidated HMW glutenin proteins. This finding questions the possibility of implementing these proteins in novel food items destined to be nontoxic for celiac disease patients.


Neurovascular manifestations in connective tissue diseases: The case of Marfan Syndrome.
Patients with connective tissue diseases (CTDs) are suspected to be at higher risk for cerebrovascular involvement, such as intracranial aneurysms, dissections and strokes, than the general population. Particularly, Marfan Syndrome (MFS) has been reported as associated with an increased risk of cerebrovascular alterations. Literature data report different prevalence of intracranial aneurysms in MFS, ranging from 4 % to 29 %, suggesting a role of genetic cause that involves the regulation of the TGF-β signaling. Ischemic and hemorrhagic strokes have been also reported in MFS, but with an estimated prevalence from 3 % to 4 %. However, the aetiology of both events appears to be reliable more to a cardiac source than to the primary connective tissue defect. Finally, the available literature suggests that MFS patients have a higher prevalence of arterial tortuosity of neck and head vessels and these findings may be related to an enhanced chance of dissection. Overall, despite of the lack of studies, we could affirm that it may exists an increased prevalence of some neurovascular findings in MFS patients. Nevertheless, further studies are required to determine the true prevalence of these features and investigate specific gene mutations involved in MFS.


Toxins in everyday life.
This article reviews the sources of exposure and health effects of common toxicants encountered by patients in primary care practice. The recognition and management of exposure to indoor and outdoor pollutants, heavy metals, pesticides, electromagnetic fields, and endocrine-disrupting chemicals are listed. A sample environmental history form is included.


Nonossifying fibromas of the distal tibia: possible etiologic relationship to the interosseous membrane.
Nonossifying fibromas (NOFs) present in a characteristic pattern in the distal tibia. Their predilection to this region and etiology remain imprecisely defined. We performed a retrospective chart review of patients between January 2003 and March 2014 for distal tibial NOFs. We then reviewed radiographs (XRs), computed tomography (CT), and magnetic resonance imaging (MRI) for specific lesion characteristics. We identified 48 distal tibia NOFs in 47 patients (31 male, 16 female; mean age 12.3 years, range 6.9-17.8). This was the second most common location in our population (30 % of NOFs), behind the distal femur (42 %). Thirty-four lesions had CT and nine had MRI. Thirty-one percent were diagnosed by pathologic fracture. Ninety-six percent of lesions were located characteristically in the distal lateral tibia by plain radiograph, in direct communication with the distal extent of the interosseous membrane on 33 of the 34 (97 %) lesions with CT available for review and all nine (100 %) with MRI. The remaining two lesions occurred directly posterior. The vast majority of distal tibial NOFs occur in a distinct anatomic location at the distal extent of the interosseous membrane, which may have etiologic implications. IV (case series).


RetroPred: A tool for prediction, classification and extraction of non-LTR retrotransposons (LINEs & SINEs) from the genome by integrating PALS, PILER, MEME and ANN.
The problem of predicting non-long terminal repeats (LTR) like long interspersed nuclear elements (LINEs) and short interspersed nuclear elements (SINEs) from the DNA sequence is still an open problem in bioinformatics. To elevate the quality of annotations of LINES and SINEs an automated tool "RetroPred" was developed. The pipeline allowed rapid and thorough annotation of non-LTR retrotransposons. The non-LTR retrotransposable elements were initially predicted by Pairwise Aligner for Long Sequences (PALS) and Parsimonious Inference of a Library of Elementary Repeats (PILER). Predicted non-LTR elements were automatically classified into LINEs and SINEs using ANN based on the position specific probability matrix (PSPM) generated by Multiple EM for Motif Elicitation (MEME). The ANN model revealed a superior model (accuracy = 78.79 +/- 6.86 %, Q(pred) = 74.734 +/- 17.08 %, sensitivity = 84.48 +/- 6.73 %, specificity = 77.13 +/- 13.39 %) using four-fold cross validation. As proof of principle, we have thoroughly annotated the location of LINEs and SINEs in rice and Arabidopsis genome using the tool and is proved to be very useful with good accuracy. Our tool is accessible at http://www.juit.ac.in/RepeatPred/home.html.