Quantifying Wheelchair Basketball Match Load: A Comparison of Heart-Rate and Perceived-Exertion Methods.
To describe the objective and subjective match load (ML) of wheelchair basketball (WB) and determine the relationship between session heart-rate (HR) -based ML and rating-of-perceived-exertion (RPE) -based ML methods. HR-based measurements of ML included Edwards ML and Stagno training impulses (TRIMPMOD), while RPE-based ML measurements included respiratory (sRPEres) and muscular (sRPEmus). Data were collected from 10 WB players during a whole competitive season. Edwards ML and TRIMPMOD averaged across 16 matches were 255.3 ± 66.3 and 167.9 ± 67.1 AU, respectively. In contrast, sRPEres ML and sRPEmus ML were found to be higher (521.9 ± 188.7 and 536.9 ± 185.8 AU, respectively). Moderate correlations (r = .629-.648, P < .001) between Edwards ML and RPE-based ML methods were found. Moreover, similar significant correlations were also shown between the TRIMPMOD and RPE-based ML methods (r = .627-.668, P < .001). That said, only ≥40% of variance in HR-based ML was explained by RPE-based ML, which could be explained by the heterogeneity of physical-impairment type. RPE-based ML methods could be used as an indicator of global internal ML in highly trained WB players.


Correction to: Relation of lowering door-to-balloon time and mortality in ST segment elevation myocardial infarction patients undergoing percutaneous coronary intervention.
The original version of this article unfortunately contained a mistake. The name of the author Keren-Lee Rozenfeld was rendered wrongly. The correct name is shown above.


Aptamer based voltammetric patulin assay based on the use of ZnO nanorods.
The authors describe an aptamer based assay for the mycotoxin patulin (PAT). A gold electrode was modified with a composite made from ZnO nanorods (ZnO-NRs) and chitosan. The ZnO-NRs was prepared by reaction with ammonia and subsequent hydrothermal growth. Its properties were characterized by X-ray diffraction, Raman spectroscopy and scanning electron microscopy. Subsequently, thiol-modified aptamers were self-assembled on AuNPs that had been electrodeposited on the surface of the modified electrode. The presence of ZnO-NRs on the electrode increases the loading with AuNPs and aptamers. It also warrants a relatively stable microenvironment for the aptamers. In the presence of PAT, it will form a complex with the aptamer on the electrode surface. This hinders electron transfer from the electrode to the redox probe hexacyanoferrate and results in reduced current, which is typically measured at 0.176 V (vs. Ag/AgCl). The concentration of PAT can be calculated from the differences in the peak current before and after incubation with PAT. The assay has a linear response in the 50 ng·mL-1 to 0.5 pg·mL-1 PAT concentration range and a 0.27 pg·mL-1 lower detection limit. The sensor is specific, reproducible, repeatable, and long-term stable. It was successfully applied to the determination of PAT in spiked juice samples. Graphical abstract Schematic representation of aptamer based detection of patulin (PAT). It is based on the fact that ZnO nanorods on the surface of the electrode increase the loading of the gold nanoparticles and the aptamers, thereby improving the electrode performance.


Frequency of 30-day readmission and its causes after percutaneous coronary intervention in acute myocardial infarction complicated by cardiogenic shock.
Survival after percutaneous coronary intervention (PCI) in acute myocardial infarction complicated by cardiogenic shock (AMI-CS) has increased over the years. Short-term readmission rates in this high-risk population remain unknown. We queried the United States (U.S.) Nationwide Readmission Database (NRD) from January 2010 to November 2014 using the International Classification of Diseases-Ninth edition, Clinical Modification (ICD-9 CM) codes to identify all patients ≥18 years readmitted within 30 days after surviving an index hospitalization for PCI in AMI-CS. Incidence, etiologies, and predictors of 30-day readmission were analyzed. Among 46,435 patients who survived to discharge after PCI in AMI-CS, 9,020 (19.4%) were readmitted within 30 days. Median time to 30-day readmission was 11 days. Cardiac conditions were the most common causes of readmission (57.8%). Heart failure was the leading readmission diagnosis (24.8%). Private insurance including HMO and self-pay were predictive of lower 30-day readmission. Among other covariates, female sex, comorbidities such as heart failure, atrial fibrillation, in-hospital complications such as major bleeding, sepsis, respiratory complications, AKI requiring dialysis, utilization of mechanical circulatory support (IABP and ECMO) were independently predictive of 30-day readmission. Trend analysis showed decline in 30-day readmission rates from 21.9% in 2010 to 17.9% in 2014 (ptrend < 0.001). In this large real-world database, one in five patients receiving PCI in AMI-CS was readmitted within 30 days after discharge. Cardiac conditions were the most common causes of readmission. Insurance type had significant influence on 30-day readmission.


Preparation of Glycan Arrays Using Pyridylaminated Glycans.
We describe the method to prepare neoglycoproteins from the conjugation of bovine serum albumin and pyridylaminated glycans. Large quantities of glycans (>1 mg) can be pyridylaminated and then converted to their 1-amino-1-deoxy derivatives by reaction with hydrogen followed by hydrazine. These pyridylaminated glycans can then be conjugated to bovine serum albumin via esterification with N-( m-maleimidobenzoyloxy)succinimide to form a neoglycoprotein, e.g., glycosylated bovine serum albumin. As a demonstration, we prepared High-mannose bovine serum albumin, which was immobilized on an activated glass slide. Then, we showed that the neoglycoprotein bind to Cy3-labeled Lens culinaris agglutinin, a mannose-specific plant lectin, as detected using an evanescent-field-activated fluorescence scanner system.


Profile and dynamics of infectious diseases: a population-based observational study using multi-source big data.
The current surveillance system only focuses on notifiable infectious diseases in China. The arrival of the big-data era provides us a chance to elaborate on the full spectrum of infectious diseases. In this population-based observational study, we used multiple health-related data extracted from the Shandong Multi-Center Healthcare Big Data Platform from January 2013 to June 2017 to estimate the incidence density and describe the epidemiological characteristics and dynamics of various infectious diseases in a population of 3,987,573 individuals in Shandong province, China. In total, 106,289 cases of 130 infectious diseases were diagnosed among the population, with an incidence density (ID) of 694.86 per 100,000 person-years. Besides 73,801 cases of 35 notifiable infectious diseases, 32,488 cases of 95 non-notifiable infectious diseases were identified. The overall ID continuously increased from 364.81 per 100,000 person-years in 2013 to 1071.80 per 100,000 person-years in 2017 (χ2 test for trend, P < 0.0001). Urban areas had a significantly higher ID than rural areas, with a relative risk of 1.25 (95% CI 1.23-1.27). Adolescents aged 10-19 years had the highest ID of varicella, women aged 20-39 years had significantly higher IDs of syphilis and trichomoniasis, and people aged ≥ 60 years had significantly higher IDs of zoster and viral conjunctivitis (all P < 0.05). Infectious diseases remain a substantial public health problem, and non-notifiable diseases should not be neglected. Multi-source-based big data are beneficial to better understand the profile and dynamics of infectious diseases.


Genome evolution in alpine oat-like grasses through homoploid hybridization and polyploidy.
Hybridization and polyploidization can radically impact genome organization from sequence level to chromosome structure. As a result, often in response to environmental change and species isolation, the development of novel traits can arise and will tend to result in the formation of homoploid or polyploid hybrid species. In this study we focus on evidence of hybridization and polyploidization by ascertaining the species parentage of the endemic alpine Helictotrichon parlatorei group. This group comprises five taxa; the diploids H. parlatorei, Helictotrichon setaceum subsp. setaceum and subsp. petzense, their putative hybrid Helictotrichon ×krischae and the hexaploid Helictotrichon sempervirens. For molecular analyses, cloned nuclear Topoisomerase VI genes of H. sempervirens and H. ×krischae were sequenced and compared with sequences of the diploids to estimate the evolutionary history in this group. In addition, detailed chromosome studies were carried out including fluorescence in situ hybridization (FISH) with 5S and 45S ribosomal and satellite DNA probes, and fluorochrome staining with chromomycin and DAPI. Two distinct types of Topoisomerase VI sequences were identified. One of them (SET) occurs in both subspecies of H. setaceum, the other (PAR) in H. parlatorei. Both types were found in H. ×krischae and H. sempervirens Karyotypes of H. parlatorei and H. setaceum could be distinguished by chromosomes with a clearly differentiated banding pattern of ribosomal DNAs. Both patterns occurred in the hybrid H. ×krischae Hexaploid H. sempervirens shares karyotype features with diploid H. parlatorei, but lacks the expected chromosome characteristics of H. setaceum, possibly an example of beginning diploidization after polyploidization. The geographic origin of the putative parental species and their hybrids and the possible biogeographical spread through the Alps are discussed.


Alkyl side-chain derivatives of sordaricin as potent antifungal agents against yeast.
Sordarin (1) was converted to 5 and 6, which showed potent antifungal activity against yeast. A series of C1-C9 alkyl side-chain derivatives was prepared, from which it was found that the optimal activity occurred with C5. A comparison of side chains with different unsaturation showed that the cis-alkene was the most active. This result suggested that the folding of the side chains might be crucial for the optimal activity.


Improvement of the energy resolution of energy dispersive spectrometers (EDS) using Richardson-Lucy deconvolution.
A serious limitation in the use of energy dispersive spectrometers (EDS) for materials characterization arises from the fact that these x-ray detectors provide a poor energy resolution compared to other x-ray techniques. This is mainly due to the broadening function generated by the electronics of the detector. However, new windowless detectors are now capable of resolving low energy peaks like Li Kα with modified electronics and show a system peak full width at half maximum (FWHM) of around 30 eV. In this paper, we investigated how the Richardson-Lucy algorithm can be used to remove, or at least attenuate, the contribution of the broadening function to experimental spectra. This method proved to be efficient in improving the energy resolution at any energy in the EDS spectrum. The resulting system peak FWHM was reduced to as low as 7-8 eV and the separation of low energy x-ray peaks were demonstrated in the low energy range of the spectra. The method was also efficient in reducing the peak broadening at higher energies (Cu Kα) and the broadening function of the detector could be experimentally determined to provide higher accuracy in predicting peak broadening. Although some critical artefacts were noted on the restored spectra, like energy shifts and intensity variations, the method explored in this work is worth to be considered for further quantification tests.


Mercury and methylmercury concentration in the feathers of two species of Kingfishers Megaceryle torquata and Chloroceryle amazona in the Upper Paraguay Basin and Amazon Basin.
Mercury (Hg) contamination remains a significant environmental concern. In aquatic ecosystems, Hg can undergo methylation, forming its organic form, methylmercury (MeHg), which bioaccumulates and biomagnifies in the food chain, ultimately reaching the top predators, including waterfowl. The objective of this study was to investigate the distribution and levels of Hg in wing feathers, with a specific focus on evaluating heterogeneity in the primary feathers of two kingfisher species (Megaceryle torquata and Chloroceryle amazona). The concentrations of total Hg (THg) in the primary feathers of C. amazona individuals from the Juruena, Teles Pires, and Paraguay rivers were 4.724 ± 1.600, 4.003 ± 1.532, and 2.800 ± 1.475 µg/kg, respectively. The THg concentrations in the secondary feathers were 4.624 ± 1.718, 3.531 ± 1.361, and 2.779 ± 1.699 µg/kg, respectively. For M. torquata, the THg concentrations in the primary feathers from the Juruena, Teles Pires, and Paraguay rivers were 7.937 ± 3.830, 6.081 ± 2.598, and 4.697 ± 2.585 µg/kg, respectively. The THg concentrations in the secondary feathers were 7.891 ± 3.869, 5.124 ± 2.420, and 4.201 ± 2.176 µg/kg, respectively. The percentage of MeHg in the samples increased during THg recovery, with an average of 95% in primary feathers and 80% in secondary feathers. It is crucial to comprehend the current Hg concentrations in Neotropical birds to mitigate potential toxic effects on these species. Exposure to Hg can lead to reduced reproductive rates and behavioral changes, such as motor incoordination and impaired flight ability, ultimately resulting in population decline among bird populations.


Coping with cryptic and defective transcripts in plant mitochondria.
Plant mitochondria are particularly prone to the production of both defective and cryptic transcripts as a result of the complex organisation and mode of expression of their genome. Cryptic transcripts are generated from intergenic regions due to a relaxed control of transcription. Certain intergenic regions are transcribed at higher rates than genuine genes and therefore, cryptic transcripts are abundantly produced in plant mitochondria. In addition, primary transcripts from genuine genes must go through complex post-transcriptional processes such as C to U editing and cis or trans splicing of group II introns. These post-transcriptional processes are rather inefficient and as a result, defective transcripts are constantly produced in plant mitochondria. In this review, we will describe the nature of cryptic and defective transcripts as well as their fate in plant mitochondria. Although RNA surveillance is crucial to establishing the final transcriptome by degrading cryptic transcripts, plant mitochondria are able to tolerate a surprising high level of defective transcripts.


Multiple urban stressors drive fish-based ecological networks in streams of Columbus, Ohio, USA.
Integrating a network perspective into multiple-stressor research can reveal indirect stressor effects and simultaneously estimate both taxonomic and functional community characteristics, thus representing a novel approach to stressor paradigms in rivers. Using six years of data from twelve streams of Columbus, Ohio, USA, the effects of nutrients (N:P), impervious surface (%IS), and sedimentation on network properties were quantified. Variability in the strength and distribution of trophic interactions was assessed by incorporating biomass into networks. All stressors impacted some properties of network topology - linkage density (average number of links per species), connectance (fraction of all possible links realized in a network), and compartmentalization (degree to which networks contain discrete sub-webs), including synergistic interactive effects between sedimentation and stream size. We also found support for antagonistic effects between (1) sedimentation and %IS and between %IS and N:P on the weighted index mean link weight, which represents the magnitude of trophic interactions among species in a network, and (2) %IS and stream size on strength standard deviation, a measure of the distribution of total magnitude of all trophic interactions per species in a network. Overall, our results point to the potential for urban stressors such as impervious surfaces and sedimentation - alone and as interactions - to decrease network complexity, compartmentalization, and stability, likely through homogenizing habitat and limiting food resources. The observation that larger streams often buffered the negative effects of these stressors suggests that restoration and other management approaches might be most beneficial in smaller headwater streams of urban catchments.


Predicting habits of vegetable parenting practices to facilitate the design of change programmes.
Habit has been defined as the automatic performance of a usual behaviour. The present paper reports the relationships of variables from a Model of Goal Directed Behavior to four scales in regard to parents' habits when feeding their children: habit of (i) actively involving child in selection of vegetables; (ii) maintaining a positive vegetable environment; (iii) positive communications about vegetables; and (iv) controlling vegetable practices. We tested the hypothesis that the primary predictor of each habit variable would be the measure of the corresponding parenting practice. Internet survey data from a mostly female sample. Primary analyses employed regression modelling with backward deletion, controlling for demographics and parenting practices behaviour. Houston, Texas, USA. Parents of 307 pre-school (3-5-year-old) children. Three of the four models accounted for about 50 % of the variance in the parenting practices habit scales. Each habit scale was primarily predicted by the corresponding parenting practices scale (suggesting validity). The habit of active child involvement in vegetable selection was also most strongly predicted by two barriers and rudimentary self-efficacy; the habit of maintaining a positive vegetable environment by one barrier; the habit of maintaining positive communications about vegetables by an emotional scale; and the habit of controlling vegetable practices by a perceived behavioural control scale. The predictiveness of the psychosocial variables beyond parenting practices behaviour was modest. Discontinuing the habit of ineffective controlling parenting practices may require increasing the parent's perceived control of parenting practices, perhaps through simulated parent-child interactions.


Synergism of the IGRs Methoprene and Pyriproxyfen Against Larval Cat Fleas (Siphonaptera: Pulicidae).
Insect growth regulators (IGRs) methoprene and pyriproxyfen are widely used as topical treatments to pets or applied to the indoor environment to control cat fleas, Ctenocephalides felis (Bouché). The toxicity of methoprene, pyriproxyfen, and combinations of both IGRs to cat flea larvae was determined. The LC50 of methoprene and pyriproxyfen applied to larval rearing medium was 0.39 and 0.19 ppm, respectively. Combinations of methoprene:pyriproxyfen in ratios of 1:1, 5:1, 10:1, and 20:1 produced LC50s of 0.06, 0.09, 0.19, and 0.13 ppm, respectively. The pyriproxyfen synergized the activity of methoprene as indicated by the combination indices (CI). The ratio of methoprene:pyriproxyfen (40:1) provided an LC50 of 0.42 ppm and the pyriproxyfen was not synergistic. Combinations of pyriproxyfen:methoprene in ratios of 5:1, 10:1, and 20:1 provided LC50s of 0.14, 0.20, 0.20 ppm, respectively, and the methoprene did not synergize the activity of pyriproxyfen. The dose-reduction indices (DRIs) indicated that the concentrations of IGRs in the combinations of methoprene:pyriproxyfen (ratios of 20:1 or less) could be reduced by at least one-third of the amount required by methoprene alone to provide similar larval mortality. Combinations of methoprene and pyriproxyfen may be effective in increasing the residual activity on pets and assist in reducing the likelihood of insecticide resistance developing to IGRs.


Mechanical Influence of Surrounding Soft Tissue on Bone Regeneration Processes: A Bone Lengthening Study.
Bone lengthening is a bone regeneration technique with multiple clinical applications. One of the most common complications of this treatment is the lack of adaptation of the surrounding soft tissue to their extension. A better understanding of the mechanobiology of the tissues involved in distraction osteogenesis would allow better control of the clinical cases. Bone lengthening treatments were performed in vivo in the metatarsus of Merino sheep, measuring the distraction forces by means of an instrumented fixator. The tissue relaxation after distraction was analyzed in this study. A viscoelastic model was also applied to distraction data to assess the mechanical behavior of the tissues during the distraction phase. Tissue relaxation is similar to other bone regeneration processes which do not imply surrounding soft tissue extension, e.g. bone transport. The effects of this tissue on distraction forces are limited to the first minutes of distraction and elongations above 4% of the original length with the protocol applied. Moreover, the surrounding soft tissue initially loses some of its viscoelasticity and subsequently suffers strain hardening from day 5 of distraction until the end of the distraction phase, day 15. Finally, anatomical changes were also evidenced in the elongated limb of our specimens.


Extranodal multiple involvement of enteropathy-type T-cell lymphoma without expression of CC chemokine receptor 7.
Enteropathy-type T-cell lymphoma (ETCL) is a rare extranodal lymphoma that tends to disseminate into the intestines and other extranodal organs. We present a case of ETCL with involvement of the lungs and kidneys and report CC chemokine receptor 7 (CCR7) expression of lymphoma cells. A 73-year-old man was admitted to the hospital with a complaint of abdominal pain. Multiple ulcers and perforations were observed in the small intestine, and partial resection of the ileum was performed. Histological examination of the resected specimen revealed diffuse proliferation of atypical large lymphoid cells. The diagnosis was ETCL with dissemination into the lungs and kidney. Lymphoma cells of the small intestine and in pleural effusion were CD3+, CD4+, CD7+, CD8-, CD25-, CD56-, CD103 +/-, and TIA-1+. Rearrangement of the T-cell receptor beta gene was detected, and human T-lymphotropic virus was not integrated. Combination chemotherapy did not result in a sustained response. The results for CCR7 expression of lymphoma cells in the lung and pleural effusion were negative. Therefore we concluded that lymphoma cells did not migrate into the lymph nodes but instead spread into the extranodal organs.


Duplication of 5q11.2----q13.1 from a familial (5;20) balanced insertion.
A 2-year-old boy with gross motor delay and few minor anomalies has a pure duplication of a small segment of chromosome 5q11.2----q13.1. A balanced insertion of this 5q segment into chromosome 20q proximal to the centromere has been found in his father, uncle, and paternal grandmother.


Evaluation of the Gold Medal Schools program.
Utah's Gold Medal Schools program supports the adoption of school policies that provide opportunities for nutritious food choices and regular physical activity. The effectiveness of Gold Medal Schools was evaluated via anthropometric measurements and dietary and physical activity surveys. The study population included first-, third-, and fifth-grade elementary school students and parents from four schools in Tooele County, UT. Two schools implemented Gold Medal Schools (intervention) and two did not (comparison). Data were collected at baseline (June 2005) and 1 year (May 2006). Body mass index (calculated as kg/m(2)) z scores increased significantly in the comparison group (0.53+/-0.38; P<0.05), but not in the intervention group (0.21+/-0.47; P=0.484), from baseline to 1 year. Children in the Gold Medal Schools cohort reported drinking fewer soft drinks per day (excluding diet drinks) at 1 year (P=0.008) and walking or biking to school more often at baseline and 1 year (P<0.001) than non-Gold Medal Schools children. While children in both groups increased the days per week they walked or biked to school, a substantial improvement was observed for the non-Gold Medal Schools students only (P<0.001). Overall, this pilot study suggests that Gold Medal Schools positively impacted body mass index z scores and health behaviors among elementary-aged students.


Noninvasive imaging markers associated with sudden cardiac death.
Sudden cardiac death (SCD) accounts for approximately 15-20% of all deaths worldwide. While the majority of SCDs occur in adults, children, and adults <35 years (<1%) may also be affected. Currently the most effective strategy for both primary and secondary prevention of SCD is the implantable cardioverter-defibrillator (ICD). However, identification of patients who will benefit from ICD implantation remains challenging. Left ventricular ejection fraction (LVEF) is the most frequent imaging parameter used to select patients for ICD implantation for primary prevention. However, LVEF has shown to be suboptimal for prediction of benefit. Non-invasive cardiac imaging permits characterization of the arrhythmogenic substrate, including dispersion of electromechanical activation, presence of myocardial scar, and cardiac innervation status. The arrhythmogenic substrate may change across the different underlying diseases. While in ischemic cardiomyopathy, differentiation and characterization of infarct core and peri-infarct zone have been shown to refine the risk stratification of patients, in non-ischemic cardiomyopathies, the substrate may be more heterogeneous and tissue characterization assessing focal and diffuse fibrosis and inflammation processes may be more relevant. Furthermore, in channelopathies, assessment of mechanical dispersion between myocardial layers may identify the patients with increased risk of ventricular arrhythmias. Finally, potential triggers of ventricular arrhythmias such as myocardial ischemia can be evaluated. The role of noninvasive imaging in the risk stratification of SCD and the selection of candidates for ICD will be discussed in this article.


Cytotoxins in non-enterotoxigenic strains of Escherichia coli isolated from feces of diarrheic calves.
We have examined the cytotoxic responses produced in HeLa and Vero cell cultures by sonicates from 15 non-enterotoxigenic (STa-, LT-) strains of E. coli, highly lethal for mice parenterally LD50 less than 3 X 10(7) CFU), which had been isolated from feces of diarrheic calves. Three types of cytotoxic responses were observed. Type 1 (five strains) consisted of enlargement, rounding and polynucleation of HeLa cells, an effect previously reported with cytotoxic necrotizing factor (CNF) in E. coli from infant and piglet enteritis. Type 2 toxicity (three strains and the control Vir strain S5) was also characterized by enlargement and polynucleation of HeLa cells, but in contrast to Type 2 effect, cells were elongated. Sonicates from the latter strains were lethal for chickens, producing the lesions previously described with Vir strains. Type 3 toxicity (two strains and the control VT strain H19), produced an extensive destruction of both Vero and HeLa cell cultures. Cytotoxic effects were completely abolished upon heating for 1 h at 60 degrees C for Type 1 and 2 extracts and at 80 degrees C for Type 3 extracts. Seroneutralization assays showed that cytotoxins of the same type were closely related antigenically. In addition, a slight cross-neutralization was observed between Type 1 (CNF) and Type 2 (Vir) toxins.


Associations of Earphone Use with Tinnitus and Anxiety/Depression.
This study aimed to investigate the association of earphone use with audiologic and psychologic factors. Korea National Health and Nutrition Examination Survey 2010-2012 data were collected for participants aged ≥12 years old with earphone use ≥1 hour/day. They were matched to a control group for age, sex, income, and education level. The relationship between earphone use and the hearing thresholds of pure-tone audiometry, tinnitus, and psychologic factors such as depression and anxiety, and other quality of life variables was analyzed using multiple logistic regression tests with complex sampling. Among the participants, 22.9% (449/1955) of earphone users and 18.1% (355/1600) of control participants had tinnitus (P < 0.001). Earphone users showed 1.27-times higher odds for tinnitus (95% confidence interval [CI] = 1.09-1.50, P = 0.003). Moreover, 6.5% (128/1955) of earphone users and 5.0% (97/1600) of control participants had anxiety and depressive symptoms (P = 0.033). Earphone users showed 1.32-times higher odds for anxiety and depressive symptoms (95% CI = 1.14-1.52, P = 0.040). Nevertheless, the hearing thresholds were comparable between earphone users and control participants. Earphone use was associated with tinnitus and anxiety or depressive symptoms.


The use of nitrocellulose immunoblots for the analysis of antigen recognition by T lymphocytes.
Antigen insolubilised on nitrocellulose is able to activate T lymphocytes following presentation by appropriate accessory cells. Furthermore, complex mixtures of antigens can be fractionated by polyacrylamide gel electrophoresis under denaturing conditions (SDS-PAGE), transferred to nitrocellulose membranes and then added to T cell proliferation assays. This allows a direct screening for recognition of individual polypeptides by monoclonal and polyclonal T cell populations, without a requirement for extensive biochemical purification and in the absence of initial serological preselection of antigen. The potential use of this technique for screening of recombinant DNA libraries for expression of antigens recognised by T cells is also illustrated. The technical aspects of T cell recognition of solid-phase antigen and its application to the identification of immunodominant epitopes with the potential to modulate T cell-mediated immunity is reviewed.


Measurement reliability of atypical language lateralisation assessed using functional transcranial Doppler ultrasound.
It is well established that some individuals present with atypical, non-left hemisphere, cerebral lateralisation for language processing. However previous studies exploring the reliability of functional blood flow responses to detect lateralised activation during speech have focused only on individuals with typical left sided dominance. Here we report test-retest and between-task reliability measures obtained with functional transcranial Doppler ultrasound in 47 participants, including 9 with atypical language presentation. Results showed good test-retest reliability in atypically lateralised individuals, even after an interval of 120 days. Between-task reliability was weaker, but still within acceptable ranges.


Tissue-wide overexpression of alpha-T-catenin results in aberrant trophoblast invasion but does not cause embryonic mortality in mice.
Transcriptional activation of CTNNA3, encoding αT-catenin, by the Y153H mutated form of the human STOX1 transcription factor was proposed to be responsible for altered fetal trophoblast invasion into the maternal endometrium during placentation in pre-eclampsia. Here we have generated a mouse model to investigate the in vivo effects of ectopic αT-catenin expression on trophoblast invasion. Histological analysis was used to determine the invasive capacities of trophoblasts from transgenic embryos, as well as proliferation rates of spongiotrophoblasts in the junctional zone. Augmented expression of αT-catenin reduced the number of invading trophoblasts but did not cause embryonic mortality. The, αT-catenin positive cells could still invade into the decidual layer and migrated as deeply as wild-type trophoblasts. Furthermore, the junctional zone is enlarged in placentas of mice overexpressing αT-catenin due to hyperproliferation of the residing spongiotrophoblasts, suggesting a pivotal role of αT-catenin levels in the control of the proliferative versus invasive state of trophoblasts during placentation. Our study provides, for the first time, in vivo data on the effects of increased levels of αT-catenin in the placenta.


Prevalence of aneuploidy, overexpressed ER, and overexpressed EGFR in random breast aspirates of women at high and low risk for breast cancer.
Breast tissue biomarkers which accurately predict breast cancer development within a 10 year period in high risk women are needed but currently not available. We initiated this study to determine 1) the prevalence of one or more breast tissue abnormalities in a group of women at high risk for breast cancer, and 2) if the prevalence of biomarker abnormalities is greater in high risk than in low risk women. Eligible high risk women were those with a first degree relative with breast cancer, prior breast cancer, or precancerous mastopathy. Low risk women were those without these or other major identifiable risk factors. Ductal cells were obtained via random fine needle aspirations and cytologically classified. Biomarkers included DNA ploidy, estrogen receptor (ER), and epidermal growth factor receptor (EGFR). The prevalence of DNA aneuploidy was 30%, overexpression of ER 10%, and overexpression of EGFR 35%, in the 206 high risk women whose median 10 year Gail risk (projected probability) of developing breast cancer was 4.5%. The prevalence of aneuploidy and overexpressed EGFR was significantly higher in the high risk women than in the 25 low risk controls (p < 0.002), whose median 10 year Gail risk was 0.7%. The difference in the prevalence of ER overexpression between high and low risk groups was not statistically significant (p = 0.095). This may be due to the low prevalence of overexpressed ER and the small number of controls. A significant difference was noted in the prevalence of one or more abnormal biomarkers between the high risk and low risk women (p < 0.001). A large prospective trial is needed to determine if one or more of these biomarkers, is predictive of breast cancer development.


Improved haplotype assembly using Xor genotypes.
Given a set of aligned fragments, haplotype assembly is the problem of finding the haplotypes from which the fragments have been read. The problem is important because haplotypes contain SNP information, which is essential to many genomic analyses such as the analysis of potential association between certain diseases and genetic variations. The current state-of-the-art haplotype assembly algorithm, HapSAT, does not exploit genotype information and only receives a read matrix as input. However, the imminent importance of haplotypes and inexpensiveness of genotype information motivate for exploiting genotype information to obtain more accurate haplotypes. In this paper, an improved haplotype assembly method, xGenHapSAT, is proposed, which exploits xor genotype information for more accurate haplotype assembly. Xor genotype information is even less expensive than full genotype information, e.g., using the Denaturing High-Performance Liquid Chromatography (DHPLC) technique. It is shown that using this inexpensively obtainable information significantly improves the accuracy of the assembled haplotypes. In addition, a new, more efficient, Max-2-SAT formulation is adopted in xGenHapSAT, which, on average, increases the speed of the algorithm. Moreover, the proposed xGenHapSAT method replaces the current state-of-the-art haplotype assembly method based on genotype information. Finally, our state-of-the-art haplotype assembly software, HapSoft, which includes both xGenHapSAT and HapSAT, is made freely available for research purposes.


The space contraction asymmetry in Michotte's launching effect.
Previous studies have found that, compared with noncausal events, spatial contraction exists between the causal object and the effect object due to the perceived causality. The present research aims to examine whether the causal object and the effect object have the same effect on spatial contraction. A modified launching effect, in which a bar bridges the spatial gap between the final position of the launcher and the initial position of the target, was adopted. Experiment 1 validates the absolute underestimation of the bar's length between the launcher and the target. Experiment 2a finds that in the direct launching effect, the perceived position of the bar's trailing edge that was contacted by the final launcher was displaced along the objects' direction of movement. Meanwhile, the perceived position of the bar's leading edge that was contacted by the initial target was displaced in opposite direction to the moving direction. The magnitude of the former's displacement was significantly larger than that of the latter, displaying a significant contraction asymmetry. Experiment 2b demonstrates that the contraction asymmetry did not result from the launcher remaining in contact with the edge of the bar. Experiment 3 indicates that contraction asymmetry showed a type of postdictive effect; that is, to some extent, this asymmetry depends on what happens after contact. In conclusion, the space between the causal object and effect object contracts asymmetrically in the launching effect, which implies that the causal object and effect object are perceived as shifting toward each other nonequidistantly in visual space.


Antigen recovery and preservation using the microwave irradiation of biological samples for transmission electron microscopy analysis.
Most studies using microwave irradiation (MWI) for the preparation of tissue samples have reported an improvement in structural integrity. However, there have been few studies on the effect of microwave (MW) on antigen preservation during sample preparation prior to immunolocalization. This report documents our experience of specimen preparation using an automatic microwave apparatus to obtain antigen preservation and retrieval. We tested the effects of MW processing vs. conventional procedures on the morphology and antigenicity of two different tissues: the brain and mammary gland, whose chemical composition and anatomical organization are quite different. We chose to locate the transcription factor PPARβ/δ using immunocytochemistry on brain tissue sections from hamsters. Antigen retrieval protocols involving MWI were used to restore immunoreactivity. We also studied the efficiency of the ultrastructural immunolocalization of both PPARγ and caveolin-1 following MWI vs. conventional treatment, on mammary gland tissue from mice at 10 days of lactation. Our findings showed that the treatment of tissue samples with MWI, in the context of a process lasting just a few hours from fixation to immunolocalization, enabled similar, or even better, results than conventional protocols. The quantification of immunolabeling for cav-1 indicated an increase in density of up to three-fold in tissues processed in the microwave oven. Furthermore, MW treatment permitted the localization of PPARβ/δ in glutaraldehyde-fixed specimens, which was impossible in the absence of MWI. This study thus showed that techniques involving the use of microwaves could largely improve both ultrastructure and immunodetection.


Doppler examination of cerebral arteries in uremic children.
Transcranial Doppler ultrasonography is a useful method for the estimation and monitoring of cerebral circulation in dialyzed patients. The aim of this study was to evaluate the effect of disease and treatment on cerebral circulation in children on maintenance hemodialysis (HD) and children prior to renal replacement therapy. We demonstrated that in uremic children blood flow velocities of the internal carotid artery (ICA), anterior cerebral artery (ACA), middle cerebral artery (MCA), and posterior cerebral artery (PCA) 120 min and 240 min from the beginning of an HD session were significantly lower than values immediately before HD. Changes in blood flow velocities of MCA and ACA during HD correlated significantly with changes in mean arterial pressure during HD. There was no correlation between changes in blood flow velocities and intradialytic changes in hematocrit values, ultrafiltration, hemoglobin concentration, and blood urea nitrogen values. Mean blood flow velocities of ICA, MCA, and PCA in euvolemic children on conservative treatment were significantly higher than after a HD session in children on maintenance HD. The factors responsible for intradialytic velocity changes of cerebral arteries in uremic children require further examination.


Frequency and phase correction of J-difference edited MR spectra using deep learning.
To investigate whether a deep learning-based (DL) approach can be used for frequency-and-phase correction (FPC) of MEGA-edited MRS data. Two neural networks (1 for frequency, 1 for phase) consisting of fully connected layers were trained and validated using simulated MEGA-edited MRS data. This DL-FPC was subsequently tested and compared to a conventional approach (spectral registration [SR]) and to a model-based SR implementation (mSR) using in vivo MEGA-edited MRS datasets. Additional artificial offsets were added to these datasets to further investigate performance. The validation showed that DL-based FPC was capable of correcting within 0.03 Hz of frequency and 0.4°of phase offset for unseen simulated data. DL-based FPC performed similarly to SR for the unmanipulated in vivo test datasets. When additional offsets were added to these datasets, the networks still performed well. However, although SR accurately corrected for smaller offsets, it often failed for larger offsets. The mSR algorithm performed well for larger offsets, which was because the model was generated from the in vivo datasets. In addition, the computation times were much shorter using DL-based FPC or mSR compared to SR for heavily distorted spectra. These results represent a proof of principle for the use of DL for preprocessing MRS data.


Internal Transcribed Spacer Region Typing of Trichophyton interdigitale Isolated from Japanese Patients.
Trichophyton interdigitale, an anthropophilic species, is one of the main causative agents of tinea unguium and tinea pedis. T. interdigitale and the zoophilic species T. mentagrophytes are morphologically and physiologically very similar. Isolates of the T. interdigitale/T. mentagrophytes complex from around the world have been classified into more than 10 internal transcribed spacer (ITS) genotypes. In this study, we isolated T. interdigitale from Japanese patients and investigated which ITS type was more common. The ITS regions of 29 clinical isolates of T. interdigitale and one clinical isolate of T. mentagrophytes were sequenced. The phylogenetic analysis of the ITS region sequences revealed that the 29 isolates of T. interdigitale belong to ITS type II of T. interdigitale. The one clinical isolate of T. mentagrophytes was in the same cluster with ITS type II* of T. mentagrophytes. One terbinafine-resistant strain of T. interdigitale also belonged to ITS type II of T. interdigitale.


Severely increased albuminuria in patients with type 2 diabetes mellitus is associated with increased subclinical atherosclerosis in femoral arteries with Na [18F]F activity as a proxy - The DETERMINE study.
Sodium [18F]fluoride (Na [18F]F) positron emission tomography imaging allows detailed visualization of early arterial micro-calcifications. This study aims to investigate atherosclerosis manifested by micro-calcification, macro-calcification, and aortic stiffness in patients with type 2 diabetes mellitus (T2DM) with and without albuminuria and severely decreased kidney function. A cohort was stratified in four groups (N = 10 per group), based on KDIGO categories (G1-5 A1-3). G1-2A1 non-diabetic controls (median [IQR] estimated glomerular filtration rate (eGFR) in mL/min/1.73 m2 91 [81-104]), G1-2A1 with T2DM (eGFR 87 [84-93], and albumin-creatinin-ratio (ACR) in mg/mmol 0.35 [0.25-0.75]), G1-2A3 with T2DM (eGFR 85 [60-103], and ACR 74 [62-122], and G4A3 with T2DM (eGFR 19 [13-27] and ACR 131 [59-304]). Na [18F]F femoral artery grading score differed significantly in the groups with the highest Na [18F]F activity in A3 groups with T2DM (G1-2A3 with T2DM 228 [100-446] and G4A3 with T2DM 198 [113-578]) from the lowest groups of the G1-2A1 with T2DM (33 [0-93]) and in G1-2A1 non-diabetic controls (75 [0-200], p = 0.001). Aortic Na [18F]F activity and femoral artery computed tomography (CT)-assessed macro-calcification was increased in G4A3 with T2DM compared with G1-2A1 with T2DM (47.5 [33.8-73.8] vs. 17.5 [8.8-27.5] (p = 0.006) and 291 [170-511] vs. 12.2 [1.41-44.3] mg (p = 0.032), respectively). Carotid-femoral pulse wave velocity (PWV)-assessed aortic stiffness was significantly higher in both A3 groups with T2DM compared with G1-2A1 with T2DM (11.15 and 12.35 vs. 8.86 m/s, respectively (p = 0.009)). This study indicates that the presence of severely increased albuminuria in patients with T2DM is cross-sectionally associated with subclinical arterial disease in terms of micro-calcification and aortic stiffness. Additional decrease in kidney function was associated with advanced macro-calcifications.


Selective cytotoxicity of squamocin on T24 bladder cancer cells at the S-phase via a Bax-, Bad-, and caspase-3-related pathways.
Annonaceous acetogenins are a group of potential anti-neoplastic agents isolated from Annonaceae plants. We purified squamocin, a cytotoxic bis-tetrahydrofuran acetogenin, from the seeds of Annona reticulata and analyzed its biologic effects on cancer cells. We showed that squamocin was cytotoxic to all the cancer lines tested. Furthermore, squamocin arrested T24 bladder cancer cells at the G1 phase and caused a selective cytotoxicity on S-phase-enriched T24 cells. It induced the expression of Bax and Bad pro-apoptotic genes, enhanced caspase-3 activity, cleaved the functional protein of PARP and caused cell apoptosis. These results suggest that squamocin is a potentially promising anticancer compound.


Vocal cord dysfunction concurrent with a nutcracker esophagus and the role of gastroesophageal reflux disease.
Psychological disorders were originally thought to be the sole cause of vocal cord dysfunction (VCD). Subsequently, other organic diseases, including structural laryngeal abnormalities, have also been reported to be associated with VCD. To describe the first patient with VCD concurrent with a nutcracker esophagus and to establish the association between VCD and gastroesophageal reflux disease (GERD) by using the Bernstein test. Symptom assessments, neuropsychiatric evaluations, fiberoptic laryngoscopy, pulmonary function tests, allergic skin prick tests, radiographs of the chest and sinuses, esophageal manometry (including 24-hour ambulatory esophageal pH monitoring), and the Bernstein test were performed. A 36-year-old woman had dyspnea, hoarseness, chest pain, and wheezes without relief for a decade. Neuropsychiatric evaluations disclosed mild depression. Fiberoptic laryngoscopy showed posterior laryngitis and paradoxical vocal cord adduction with audible inspiratory stridor. Pulmonary function tests showed attenuation of the inspiratory limb with notching in both flow-volume loops and a mid-vital capacity expiratory to inspiratory flow ratio of 4. All the symptoms except chest pain were improved dramatically by speech therapy and empirical treatment for GERD. Esophageal manometry revealed a nutcracker esophagus; 24-hour ambulatory esophageal pH monitoring demonstrated multiple short reflux episodes. The Bernstein test was conducted, and all the manifestations were reproduced with 0.1 N hydrochloric acid but not with isotonic sodium chloride infusion. This is the first human case report confirming that GERD can trigger an acute attack of VCD and may induce chest pain as a nutcracker esophagus in patients with VCD. It strengthens this association and expands our knowledge of diverse manifestations of this clinical entity.


Quantifying intra- and inter-species contact rates at supplemental feeding sites in Ethiopia to inform rabies maintenance potential of multiple host species.
Rabies, a multi-host pathogen responsible for the loss of roughly 59,000 human lives each year worldwide, continues to impose a significant burden of disease despite control efforts, especially in Ethiopia. However, how species other than dogs contribute to rabies transmission throughout Ethiopia remains largely unknown. In this study, we quantified interactions among wildlife species in Ethiopia with the greatest potential for contributing to rabies maintenance. We observed wildlife at supplemental scavenging sites across multiple landscape types and quantified transmission potential. More specifically, we used camera trap data to quantify species abundance, species distribution, and intra- and inter-species contacts per trapping night over time and by location. We derived a mathematical expression for the basic reproductive number (R0 ) based on within- and between-species contract rates by applying the next generation method to the susceptible, exposed, infectious, removed model. We calculated R0 for transmission within each species and between each pair of species using camera trap data in order to identify pairwise interactions that contributed the most to transmission in an ecological community. We estimated which species, or species pairs, could maintain transmission ( R 0 > 1 ${R_0} > 1$ ) and which species, or species pairs, had contact rates too low for maintenance ( R 0 < 1 ${R_0} < 1$ ). Our results identified multiple urban carnivores as candidate species for rabies maintenance throughout Ethiopia, with hyenas exhibiting the greatest risk for rabies maintenance through intra-species transmission. Hyenas and cats had the greatest risk for rabies maintenance through inter-species transmission. Urban and peri-urban sites posed the greatest risk for rabies transmission. The night-time hours presented the greatest risk for a contact event that could result in rabies transmission. Overall, both intra- and inter-species contacts posed risk for rabies maintenance. Our results can be used to target future studies and inform population management decisions.


Chemical complementarity between immune receptor CDR3s and IDH1 mutants correlates with increased survival for lower grade glioma.
Focusing on highly specific aspects of the immune response is likely to answer a number of basic questions, and in some cases even resolve basic contradictions, in cancer immunology. For example, there are many cases, where chronic inflammation is associated with cancer development, and many other cases where an immune response represents an anticancer process. In this study, using bioinformatics algorithms, we examined the chemical relationships between the amino acid sequences of the complementarity-determining region-3 (CDR3) represented by immune receptors associated with lower grade glioma and isocitrate dehydrogenase-1 (IDH1) mutants. In particular, we developed a chemical complementarity scoring approach to classify tumors based on the complementarity of CDR3s and mutant IDH1 amino acids, relying on net charge per residue and hydropathy parameters. There was a strong correlation between the increased survival in low-grade glioma (LGG) and complementarity of IDH1 mutants to the CDR3 domain of the T-cell receptor beta chain (TRB). Similar results were obtained for TRB CDR3s and NRAS mutants in melanoma. Furthermore, the clear connection between increased survival rates and immune receptor-IDH1 mutant complementarities may also, partially, explain the better LGG prognosis for patients with IDH1 mutants.


Mediators of vascular and parenchymal mechanisms in secondary brain damage.
Several putative mediators of vasogenic brain edema will be considered with respect to the following criteria: 1) their effect on blood-brain barrier (BBB) permeability, 2) their vasomotor actions which may increase driving forces for transmural bulk flow, 3) their influence on edema formation, 4) their actual tissue concentration in pathological states, and 5) the therapeutic results after specific treatment. Bradykinin (BK) can induce brain edema by increasing BBB permeability to small solutes and enhancing blood pressure in the microcirculation due to arterial dilatation and venous constriction. Its interstitial concentration is enhanced after experimental trauma. Since kallikrein inhibitors reduce brain swelling all criteria favour BK as a mediator of vasogenic edema. Arachidonic acid (AA) opens BBB also for large tracers but exerts only small vasomotor effects. The edema formation is associated with an increase of the AA concentration in the interstitial space. However, convincing therapeutic results on inhibition of AA are still lacking. In addition to the formation of vasogenic edema AA has been found to induce cytotoxic edema. From experiments dealing with the vasomotor effects Ellis et al. (Am J Physiol 255: H397-H400, 1988) concluded an interaction of BK and AA in brain injury. However, our own results do not favour this hypothesis since we found divergent vasomotor and permeability effects of BK and AA. Histamine (HA) opens BBB unspecifically and dilates cerebral vessels, mechanisms by which edema formation can be explained.(ABSTRACT TRUNCATED AT 250 WORDS)


Congenital alopecia, seizures, and psychomotor retardation in three siblings.
Three siblings, devoid of hair at birth, had an unusual autosomal recessive disorder characterized by universal congenital alopecia, microcephaly, seizures, psychomotor retardation, and severe growth failure. Metabolic and chromosome studies were normal. Skin biopsies disclosed immature hair follicles, some of which were filled with keratotic material but had no hair shafts. Neuropathologic features included cerebral cortical hypoplasia, neuronal depletion, and microcalcifications. The familial occurrence of universal congenital alopecia conjoined with nonprogressive central nervous system abnormalities in this and other kindreds defines a nosologic group of neurocutaneous disorders in which congenital alopecia is the solitary cutaneous manifestation.


Effect of natural rubber processing sludge on the degradation of crude oil hydrocarbons in soil.
Crude oil-polluted soil (five parts of weathered crude oil per 100 parts of soil; equivalent to 50,000 mg oil kg(-1) soil) samples were slurried in deionised water (300% of the water retention capacity of the soil) and treated with various amounts of natural-rubber processing sludge (nitrogen content 62.15 mg kg(-1) and phosphorus contents 8.75 mg kg(-1)) in a well-stirred, continuously-aerated tank at 29 degrees C. Changes in the total hydrocarbon content of the soil sample were determined, using a spectrophotometric technique, as a function of time. The extent of crude oil degradation was markedly higher (by up to 100%) in the sludge-treated soil than in the untreated soil sample. The efficiency of biodegradation of the crude oil hydrocarbons using the slurry-phase technique was compared with that of solid-phase technique.


Neural stem cells grafts decrease neural apoptosis associated with caspase-7 downregulation and BDNF upregulation in rats following spinal cord hemisection.
Transplantation of neural stem cells (NSCs) into lesioned spinal cord demonstrated a beneficial effect for neural repair, the underlying mechanism, however, remains to be elusive. Here, we showed that NSCs, possessing the capacity to differentiate toward into neurons and astrocytes, exhibit a neuroprotective effect by anti-apoptosis mechanism in spinal cord hemi-transected rats despite it did not improve behavior. Intravenous NSCs injection substantially upregulated the level of BDNF mRNA but not its receptor TrkB in hemisected spinal cord, while caspase-7, a downstream apoptosis gene of caspase-3, has been largely down-regulated. TUNEL staining showed that the number of apoptosis cells in injured spinal cord decreased significantly, compared with seen in rats with no NSCs administration. The present finding therefore provided crucial evidence to explain neuroprotective effect of NSCs grafts in hemisected spinal cord, which is associated with BDNF upregulation and caspase-7 downregulation.


Simulation in undergraduate paediatrics: a cluster-randomised trial.
Medical students lack confidence in recognising, assessing and managing unwell patients, particularly children. Our aim was to evaluate the impact of a 1-day novel paediatric simulation course on medical students' ability to recognise and assess sick children, and to evaluate medical students' views on the use of simulation in child health teaching. We conducted a cluster-randomised trial with a mixed-methods design. Students were cluster randomised into the intervention (simulation) group or control group (standard paediatric attachment). Students in the intervention group attended a 1-day simulation course during the last week of their attachment. The primary outcome measure was students' self-reported ability and confidence in recognising, assessing and managing sick children. There were 61 students in the study: 32 in the intervention group and 29 in the control group. Self-assessed confidence in recognising, assessing and managing a sick child was higher after the simulation course, compared with controls (p < 0.001). Six key themes were identified, including: increased confidence in emergency situations; the value of learning through participation in 'real-life' realistic scenarios in a safe environment; and an appreciation of the importance of human factors. Students found the simulation useful and wanted it offered to all undergraduates during child health attachments. A 1-day simulation course improves medical students' confidence in assessing and managing unwell children, and is highly valued by students. It could be used to complement undergraduate teaching on the management of sick children. Further studies are needed to evaluate its impact on real-life clinical performance and confidence over time. Students lack confidence in managing unwell patients, particularly children.


Effects of recombinant plasminogen activator inhibitor type 1 on fibrinolysis in vitro and in vivo.
A biologically active recombinant PAI-1 (rPAI-1) was evaluated for its effects on clot lysis in vitro and in vivo. At concentrations of 0.5 to 10 micrograms/ml, the rPAI-1 significantly prolonged the time to lysis of rabbit euglobulin clots both in the presence and absence of exogenous tissue plasminogen activator. To examine the effects of PAI in vivo, we infused rPAI-1 into conscious rabbits after i.v. injection of homogenized fibrin clots, and assessed fibrinolysis by measuring the appearance of d-dimer fibrin degradation products (FDP). Plasma fibrinolytic activity, PAI activity and antigen were also measured in plasma samples taken during and after infusion of rPAI-1. In control rabbits, endogenous fibrinolytic activity resulted in a significant and continual generation of FDP, reaching 32.6 +/- 13.6 ng/ml 90 minutes after fibrin injection. Infusion of 1, 2, or 5 micrograms/kg/min rPAI-1 led to dose dependent increases in PAI activity and antigen, while FDP levels at 90 minutes were only 8.8 +/- 2.9, 5.7 +/- 2.4, and 0.3 +/- 0.3 ng/ml, respectively. Complete inhibition of fibrinolysis was observed with 10 micrograms/kg/min rPAI-1. These studies directly demonstrate that increases in PAI-1 impair the fibrinolytic system, and support indirect observations of an association between elevated levels of this protein and thromboembolic diseases.


Primary study of lymph node metastasis-related serum biomarkers in breast cancer.
The aim of this study was to detect the pretreatment serum protein profiles of breast cancer patients by mass spectrometry (MS) to screen candidate tumor biomarkers, which will supply a simple, accurate, and minimally invasive method to predict the axillary lymph node metastasis of breast cancer. We used magnetic bead-based weak cation-exchange chromatography followed by matrix-assisted laser desorption and ionization time-of-flight MS to detect proteins in the sera of 54 cases of axillary node-negative breast cancer, 47 cases of axillary node-positive breast cancer, and 101 healthy controls. The protein profiles were analyzed to screen tumor biomarkers and lymph node metastasis-associated proteins to establish and verify a diagnostic model. Comparison of the protein profiles between the two cancer groups resulted in a total of 111 discriminate m/z peaks that were associated with breast cancer. Furthermore, 40 discriminate m/z peaks were detected between breast cancer patients with and without axillary node metastases. Four protein m/z peaks at 5,643, 4,651, 2,377, and 2,240 were used to construct a diagnosis model, and cross-validation indicated that breast cancer with and without axillary node metastasis was identified with 87.04% sensitivity (47/54), 87.23% specificity (41/47), and 87.13% accuracy (88/101). These proteins could potentially be used as predictive biomarkers to distinguish between breast cancer patients with or without lymph node metastasis.


Severe hypermotility during sleep in treatment of cataplexy with clomipramine.
A case of narcolepsy with marked cataplexy is described in detail. Clomipramine hydrochloride effectively controlled the cataplexy and partially alleviated the daytime sleep attacks. However, clomipramine treatment resulted in episodes of severe motor hyperactivity during sleep, which were most intense during rapid eye movement sleep.


Patient reported outcomes and ankle plantarflexor muscle performance following gastrocnemius recession for Achilles tendinopathy: A prospective case-control study.
Prospective studies to guide the application of a gastrocnemius recession for Achilles tendinopathy are limited. Our aim was to prospectively evaluate patient reported outcomes and muscle performance. Patients with unilateral recalcitrant Achilles tendinopathy who received an isolated gastrocnemius recession (n=8) and a healthy control group (n=8) were included. Patient reported outcomes, ankle power during walking and stair ascent, and the heel rise limb symmetry index (total work) were collected. Improvements in pain and self-reported function were observed (six months and two years). Sport participation scores reached 92% by two years. Patients demonstrated lower ankle power during stair ascent and decreased limb symmetry during heel rise six months following treatment (p≤.02). Study findings regarding long-term improvements in patient pain, self-reported function and sport participation, and early preservation of ankle function during walking, can help refine patient selection, anticipated outcomes, and rehabilitation strategies.


X-chromosomal localization of mammalian Y-linked genes in two XO species of the Ryukyu spiny rat.
Ryukyu spiny rats (genus Tokudaia), which are endemic to the central part of the Nansei Shoto archipelago in Japan, have unique karyotypes with odd numbers of chromosomes and no cytologically recognizable Y chromosome. The chromosome numbers of Tokudaia osimensis from Amamioshima and of Tokudaia sp. from Tokunoshima are 2n = 25 and 2n = 45, respectively, with a putative single X chromosome. The mouse X probe hybridized to the unpaired X chromosome, except for the distal part of the short arm in a female specimen of T. osimensis and in one male and one female of Tokudaia sp. Fluorescence in situ hybridization with the Tspy (testis-specific protein, Y-encoded) gene from both male and female cells of Tokudaia sp. by PCR localized Tspy to the distal part of the long arm of the X chromosome. Another Y-related gene, Zfy, from Tokudaia sp. was also localized to the same region in both species. Although the Sry gene is absent in this species, the present results suggest that the Y-chromosome segment carrying functional Y-linked genes, such as Tspy and Zfy, is translocated onto the distal part of the long arm of the X chromosome.


GABAergic system of the pineal organ of an elasmobranch (Scyliorhinus canicula): a developmental immunocytochemical study.
The present immunocytochemical study provides evidence of a previously unrecognized, rich, gamma-aminobutyric acid (GABA)-ergic innervation of the pineal organ in the dogfish (Scyliorhinus canicula). In this elasmobranch, the pineal primordium is initially detected at embryonic stage 24 and grows to form a long pineal tube by stage 28. Glutamic acid decarboxylase (GAD)-immunoreactive (-ir) fibers were first observed at stage 26, and by stage 28, thin GAD-ir fibers were detectable at the base of the pineal neuroepithelium. In pre-hatchling embryos, most fibers gave rise to GAD-ir boutons that were localized in the basal region of the neuroepithelium, although a smaller number of labeled terminals ascended to the pineal lumen. A few pale GAD-ir perikarya were observed within the pineal organ of stage 29 embryos, but GAD-ir perikarya were not observed at other developing stages or in adults. In contrast, GABA immunocytochemistry revealed the presence of GABAergic perikarya and fibers in the pineal organ of late stage embryos and adults. Although high densities of GABAergic cells were observed in the paracommissural pretectum, posterior tubercle, and tegmentum of dogfish embryos (regions previously demonstrated to contain pinealopetal cells), the presence of GABA-ir perikarya in the pineal organ strongly suggests that the rich GABAergic innervation of the elasmobranch pineal organ is intrinsic. This contrasts with the central origin of GABAergic fibers in the pineal gland of some mammals.


Late diastolic mitral regurgitation studied by pulsed Doppler echocardiography.
To evaluate the usefulness of pulsed Doppler echocardiography in assessing late diastolic mitral regurgitation (MR) and to clarify the pathophysiology of MR, 226 consecutive patients who had undergone left ventriculography were studied. By investigating blood flow patterns at the left atrial outflow tract, late diastolic disturbed flow suggesting MR was detected in 15 patients (7%), including 10 (4%) with positive left ventriculographic findings. Among these 15 patients, 14 (93%) had atrial fibrillation and had late diastolic MR in the cardiac cycle with prolonged RR interval. The limitation in number of cardiac cycles that could be analyzed and the rapid heart rate eliminating appearance of the beat with prolonged RR interval may be the reasons for the paucity of late diastolic MR by left ventriculography. Ten patients (66%) with late diastolic MR, including 1 with sinus rhythm, had aortic regurgitation, 3 had high-grade systolic MR and 2 had atrial septal defect. Simultaneous recording of pulmonary artery wedge pressure and left ventricular pressure in 3 patients showed a reversal of pressure gradient in late diastole when the RR interval was prolonged. In conclusion, pulsed Doppler echocardiography was useful for detecting late diastolic MR and in reducing overestimation of systolic MR in left ventriculography induced by erroneous addition of late diastolic MR. The difference of the incidence of this flow between left ventriculography and Doppler examination indicated that this flow depends primarily on heart rate and may come and go in a patient.


A novel quantitative targeted analysis of X-chromosome inactivation (XCI) using nanopore sequencing.
X-chromosome inactivation (XCI) analyses often assist in diagnostics of X-linked traits, however accurate assessment remains challenging with current methods. We developed a novel strategy using amplification-free Cas9 enrichment and Oxford nanopore technologies sequencing called XCI-ONT, to investigate and rigorously quantify XCI in human androgen receptor gene (AR) and human X-linked retinitis pigmentosa 2 gene (RP2). XCI-ONT measures methylation over 116 CpGs in AR and 58 CpGs in RP2, and separate parental X-chromosomes without PCR bias. We show the usefulness of the XCI-ONT strategy over the PCR-based golden standard XCI technique that only investigates one or two CpGs per gene. The results highlight the limitations of using the golden standard technique when the XCI pattern is partially skewed and the advantages of XCI-ONT to rigorously quantify XCI. This study provides a universal XCI-method on DNA, which is highly valuable in clinical and research framework of X-linked traits.


The binding of cellulase variants to dislocations: a semi-quantitative analysis based on CLSM (confocal laser scanning microscopy) images.
Binding of enzymes to the substrate is the first step in enzymatic hydrolysis of lignocellulose, a key process within biorefining. During this process elongated plant cells such as fibers and tracheids have been found to break into segments at irregular cell wall regions known as dislocations or slip planes. Here we study whether cellulases bind to dislocations to a higher extent than to the surrounding cell wall. The binding of fluorescently labelled cellobiohydrolases and endoglucanases to filter paper fibers was investigated using confocal laser scanning microscopy and a ratiometric method was developed to assess and quantify the abundance of the binding of cellulases to dislocations as compared to the surrounding cell wall. Only Humicola insolens EGV was found to have stronger binding preference to dislocations than to the surrounding cell wall, while no difference in binding affinity was seen for any of the other cellulose variants included in the study (H. insolens EGV variants, Trichoderma reesei CBHI, CBHII and EGII). This result favours the hypothesis that fibers break at dislocations during the initial phase of hydrolysis mostly due to mechanical failure rather than as a result of faster degradation at these locations.


On-line monitoring of microg/L levels of haloacetic acids using ion chromatography with post-column nicotinamide reaction and fluorescence detection.
A method for measuring the concentrations of the five regulated haloacetic acids (HAA5) in drinking water is reported. This method uses ion chromatography to separate HAA5 species, followed by post-column reaction with nicotinamide and detection of the fluorescent products. The result of method detection limit, accuracy, precision, linearity and interference studies are reported. The on-line monitoring method is compared directly to USEPA 552.3 in Memphis drinking water. Though not meant to replace the USEPA 552.3 for compliance monitoring, the proposed method does offer attractive alternatives considering the ease of automation and application of on-line monitoring directly from drinking water distribution systems.


Intraplantar injection of glutamate evokes peripheral adenosine release in the rat hind paw: involvement of peripheral ionotropic glutamate receptors and capsaicin-sensitive sensory afferents.
Glutamate receptors have been identified on the peripheral terminals of both primary sensory afferents and sympathetic post-ganglionic neurons, and activation of these receptors produces peripheral sensitization and enhances nociception. Adenosine is an endogenous agent that has a regulatory effect on pain. In brain and spinal cord, adenosine release can be promoted by excitatory amino acids. In the present study, we used in vivo microdialysis to determine whether glutamate also can release adenosine in peripheral tissues. Rats were anesthetized with pentobarbital and microdialysis probes were implanted into the subcutaneous tissue of the plantar aspect of the rat hind paw. Subcutaneous injection of glutamate (50 microL, 0.3-100 micromol) evoked a short-lasting adenosine release immediately following drug injection. Co-administration of either the N-methyl-D-aspartate (NMDA) receptor antagonist, dizocipine maleate (MK-801, 1 nmol) or the non-NMDA receptor antagonist, 6-cyano-7-nitroquinoxaline (CNQX, 10 nmol) with glutamate blocked such release, suggesting an involvement of peripheral ionotropic glutamate receptors in this response. Systemic pre-treatment with capsaicin, a neurotoxin selective for unmyelinated sensory afferents, significantly reduced glutamate-evoked peripheral adenosine release, but release was not affected by systemic pre-treatment with 6-hydroxydopamine, a neurotoxin selective for sympathetic nerve efferents. Neither MK-801 nor CNQX blocked 5% formalin-evoked adenosine release, suggesting adenosine release by formalin is not secondary to ionotropic glutamate receptor activation. We conclude that administration of glutamate evokes peripheral adenosine release, and that peripheral ionotropic glutamate receptors on unmyelinated sensory afferents are involved in such release. The released adenosine may provide a negative feedback control on nociception.


New insights on molecular internalization and drug delivery following plasma jet exposures.
In this study, we evaluated cold atmospheric plasmas as a physical drug delivery tool for human cervical cancer HeLa cells and murine breast carcinoma 4T1 cells. Different cell exposure protocols - plasma jet, plasma treated medium, and combinations of plasma-induced electric field and plasma treated medium- have been proposed and assessed to provide new insight on plasma-induced uptake mechanism. Cell culture medium composition and volume are key parameters to achieve an efficient molecular uptake. The plasma device enabled the delivery of molecules having 150 kDa-size into 4T1cells. For the first time to our knowledge, substance uptake kinetics after plasma treatment were investigated. The percentage of positive cells for propidium iodide and an anti-cancer agent, doxorubicin, was higher when the drugs were added a few minutes after treatment. The Plasma treated medium was not found to be as efficient as direct plasma treatment in 4T1 cells while allowing an efficient delivery in HeLa cells. Uptake levels as high as 39.3 ± 2.9% and 40.1 ± 9.5% for HeLa and 4 T1 cells respectively were achieved for optimized operating conditions, for which the viability of the cells was not severely affected. We also observed that plasma treatment induced the formation of actin stress fibers into cells revealing a mechanical stress.


The effect of an ICU liaison nurse-led family-centred transition intervention program in an adult ICU.
ICUs follow a restrictive companionship policy, especially after the COVID-19 outbreak. This strategy often limits the time families spend with patients, hinders their knowledge and skills in caregiving, and the sudden transfer of ICU patients to assist with disease monitoring and daily care can be very stressful for families. It is beneficial to use the transition period of transferring ICU patients to help families adjust to the role of caregiver. To develop and implement a patient- and family-centered transitional care intervention plan for patients transferred to the ICU. Prospective, pretest and posttest design. The experimental group received an individualized family-centered transition plan led by the ICU liaison nurse that included 1) communication with health care professionals; distribution of a transfer booklet; 2) identification of nursing issues and communication with the ward nurse; invitation of family members to participate in the patient's rehabilitation; 3) follow-up instruction on bedside range of motion exercises; and provision of a patient rehabilitation diary. Patients in the control group received only routine care. Data were collected using the General Information Questionnaire, Family Satisfaction with ICU Patients (FS-ICU), the Family Relocation Stress Scale (FRSS), and the Stanford Acute Stress Response Questionnaire (SASRQR). After the intervention, the total family satisfaction score of ICU patients in the experimental group was significantly higher than that of the control group (87.18 ± 8.38 vs 78.74 ± 10.63, p<0.001), and the satisfaction with the care and information provided was significantly higher in the experimental group compared to the control group (p < 0.001), with no significant difference between the two groups in terms of satisfaction with decision making (p>0.001). The level of relocation stress of patients' families was significantly lower in the experimental group compared to the control group after the intervention (p < 0.001). And there was no statistically significant difference between the two groups in terms of patients' acute stress disorder scores (p>0.001). The implementation of a family-involved transition care programme significantly improved the satisfaction of ICU patients' families and reduced the stress of relocation for patients' families. Focusing on the transition of ICU patients to ensure continuity of critical care and improve the quality of care for ICU patient transfers can be accomplished through a family-centered transition care plan led by the ICU liaison nurse.


Cancer in Lockdown: Impact of the COVID-19 Pandemic on Patients with Cancer.
The lockdown measures of the ongoing COVID-19 pandemic have disengaged patients with cancer from formal health care settings, leading to an increased use of social media platforms to address unmet needs and expectations. Although remote health technologies have addressed some of the medical needs, the emotional and mental well-being of these patients remain underexplored and underreported. We used a validated artificial intelligence framework to conduct a comprehensive real-time analysis of two data sets of 2,469,822 tweets and 21,800 discussions by patients with cancer during this pandemic. Lung and breast cancer are most prominently discussed, and the most concerns were expressed regarding delayed diagnosis, cancellations, missed treatments, and weakened immunity. All patients expressed significant negative sentiment, with fear being the predominant emotion. Even as some lockdown measures ease, it is crucial that patients with cancer are engaged using social media platforms for real-time identification of issues and the provision of informational and emotional support.


Intraocular lens power calculation using ray tracing following excimer laser surgery.
To evaluate intraocular lens (IOL) power calculation using ray tracing in patients presenting with cataract after excimer laser surgery. Ten eyes of seven consecutive patients who presented for cataract surgery following excimer laser treatment without any pre-refractive biometry data were enrolled in this prospective clinical study. Preoperatively, IOL power calculation was performed using a ray tracing software called OKULIX. Keratometry data (C-Scan) were imported and axial length (IOLMaster) was entered manually. Accuracy of IOL power calculation was investigated by subtracting attempted and achieved spherical equivalent. Mean spherical equivalent was -3.51+/-2.77 D (range -10.38 to -0.5 D) preoperatively and -1.01+/-1.08 D (range -2.5 to +0.75 D) postoperatively. Mean error was 0.31+/-0.84 D, mean absolute error was 0.74+/-0.46 D, and IOL calculation errors ranged from -1.39 to +1.47 D. A total of 40% of eyes were within +/-0.5 D, 70% within +/-1.0 D, and 100% within +/-1.5 D. Three eyes with corneal radii over 10 mm showed calculation errors exceeding +/-1.0 D. Mean best-corrected visual acuity increased from 20/60 to 20/30 postoperatively. IOL power calculation after excimer laser surgery can be difficult, especially when pre-refractive keratometry values are not available. In these cases, ray tracing combined with corneal topography measurements provides reliable and satisfactory postoperative results. However, it is advisable to be careful when calculating IOL power for eyes with corneal radii exceeding 10 mm because of slightly higher prediction errors.


New developments in neonatal respiratory management.
Respiratory distress syndrome (RDS) is the major cause of respiratory failure in preterm infants due to immature lung development and surfactant deficiency. Although the concepts and methods of managing respiratory problems in neonates have changed continuously, determining appropriate respiratory treatment with minimal ventilation-induced lung injury and complications is crucially important. This review summarizes neonatal respiratory therapy's advances and available strategies (i.e., exogenous surfactant therapy, noninvasive ventilation, and different ventilation modes), focusing on RDS management.


Testing a Novel Disposable Aqueous Humor Collector: An Approach to Improve Safety, Accuracy, and Efficiency.
Aqueous humor (AH) is a useful biofluid for differential diagnosis and disease monitoring in a variety of ophthalmologic conditions. There are no commercially available medical devices specially designed for AH sampling. To improve the safety, accuracy, efficiency, and convenience of anterior chamber (AC) paracentesis, a novel vacuum-based disposable AH collector with a draw volume of 50 μL was designed. The safety and performance of this novel medical device was tested in New Zealand white rabbits. A commonly used 25G 1 mL syringe was used as the control device with target AH collection volume also intentionally set at 50 μL. The 36 eyes included in the study (18 rabbits) were divided into collector (R), collector (L), syringe (R), and syringe (L) subgroups, and each included 9 eyes (R/L indicates the paracentesis was performed by right/left hand). The mean AH volume collected by the collector (R), collector (L), syringe (R), and syringe (L) subgroups were 46.66 ± 3.37 (range: 39.20-50.40), 48.71 ± 2.88 (range: 45.00-53.60), 85.11 ± 18.70 (range: 64.00-123.50), and 80.68 ± 20.87 (range: 36.8-115.8) μL, respectively. The mean absolute deviation from the target volume and mean operation time of the collector subgroups were significantly lower than the syringe subgroups. Seidel tests revealed no AH leakage in any of the tested eyes. This study revealed that this novel AH collector facilitates one-handed AC paracentesis and accurate AH sampling, and appeared to be safer and more efficient than the traditional syringe-based techniques.


PEG-mesoporous material modified by superparamagnetic nanoparticles as a delivery system of cefotaxime.
The fight against infectious bacterial diseases has been taking major steps forward by designing new nanodrugs and nanoscale biological carriers with unique physicochemical properties. Here, polyethylene glycol (PEG)-silica mesoporous material/superparamagnetic CuFe2O4 nanoparticle nanocomposite was synthesized using a facile chemical approach, and proposed as a carrier for the delivery of cefotaxime (CTX) drug. The resulting nanocomposite material and nanocomposite CTX drug carrier were characterized by different techniques. Antibacterial activity of the nanocomposite drug carrier was investigated against E.coli and S. aureus bacteria, and compared with that of commercial CTX drug. It was found that the minimum inhibitory concentrations of the nanocomposite drug carrier (1.25 and 5 μg/mL) were significantly less than those of the commercial drug (10 and 80 μg/mL) against the bacteria. Furthermore, enhanced performance of the nanocomposite CTX carrier for both the Gram-negative and Gram-positive bacteria was evidenced.


Peroxynitrite is a potent inhibitor of NF-{kappa}B activation triggered by inflammatory stimuli in cardiac and endothelial cell lines.
Peroxynitrite is a potent oxidant and nitrating species proposed as a direct effector of myocardial damage in numerous cardiac pathologies. Whether peroxynitrite also acts indirectly, by modulating cell signal transduction in the myocardium, has not been investigated. Therefore, we examined a possible role for peroxynitrite on the activation of NF-kappaB, a crucial pro-inflammatory transcription factor, in cultured H9C2 cardiomyocytes. H9C2 cells were stimulated with tumor necrosis factor-alpha or lipopolysaccharide following a brief (20-min) exposure to peroxynitrite. NF-kappaB activation (phosphorylation and degradation of its inhibitor IkappaBalpha, nuclear translocation of NF-kappaB p65, and NF-kappaB DNA binding) triggered by lipopolysaccharide or tumor necrosis factor-alpha was abrogated by peroxynitrite. Peroxynitrite also inhibited NF-kappaB in two human endothelial cell lines activated with tumor necrosis factor-alpha or interleukin-1beta. These effects were related to oxidative but not nitrative chemistry and were still being observed while nitration was suppressed by epicatechin. The mechanism of NF-kappaB inhibition by peroxynitrite was a complete blockade of phosphorylation and activation of the upstream kinase IkappaB kinase (IKK) beta, required for canonical, pro-inflammatory NF-kappaB activation. At the same time, peroxynitrite activated phosphorylation of NF-kappaB-inducing kinase and IKKalpha, considered as part of an alternative, noncanonical NF-kappaB activation pathway. Suppression of IKKbeta-dependent NF-kappaB activation translated into a marked inhibition of the transcription of NF-kappaB-dependent genes by peroxynitrite. Thus, peroxynitrite has a dual effect on NF-kappaB, inhibiting canonical IKKbeta-dependent NF-kappaB activation while activating NF-kappaB-inducing kinase and IKKalpha phosphorylation, which suggests its involvement in an alternative pathway of NF-kappaB activation. These findings offer new perspectives for the understanding of the relationships between redox stress and inflammation.


Location of motoneurons innervating the musculus sphincter cloacae in the domestic fowl.
Horseradish peroxidase was injected into the musculus (m.) sphincter cloacae through the mucosa from the lumen side of the cloaca in the domestic fowl. Labeled neurons were found in the ventrolateral area of the ventral horn from the lumbosacral segments 7-9. The craniocaudal distribution of labeled neurons overlapped with that of hindlimb motoneurons. The m. sphincter cloacae is innervated by motoneurons sending axons to the sphincter cloacae either through the connexus caudalis branching from the caudal coxal nerve of the sacral plexus or through the lateral caudal nerve arising from the pudendal plexus.


Structural, energetic, and mechanical perturbations in rhodopsin mutant that causes congenital stationary night blindness.
Several point mutations in rhodopsin cause retinal diseases including congenital stationary night blindness and retinitis pigmentosa. The mechanism by which a single amino acid residue substitution leads to dysfunction is poorly understood at the molecular level. A G90D point mutation in rhodopsin causes constitutive activity and leads to congenital stationary night blindness. It is unclear which perturbations the mutation introduces and how they can cause the receptor to be constitutively active. To reveal insight into these mechanisms, we characterized the perturbations introduced into dark state G90D rhodopsin from a transgenic mouse model expressing exclusively the mutant rhodopsin in rod photoreceptor cells. UV-visible absorbance spectroscopy revealed hydroxylamine accessibility to the chromophore-binding pocket of dark state G90D rhodopsin, which is not detected in dark state wild-type rhodopsin but is detected in light-activated wild-type rhodopsin. Single-molecule force spectroscopy suggested that the structural changes introduced by the mutation are small. Dynamic single-molecule force spectroscopy revealed that, compared with dark state wild-type rhodopsin, the G90D mutation decreased energetic stability and increased mechanical rigidity of most structural regions in the dark state mutant receptor. The observed structural, energetic, and mechanical changes in dark state G90D rhodopsin provide insights into the nature of perturbations caused by a pathological point mutation. Moreover, these changed properties observed for dark state G90D rhodopsin are consistent with properties expected for an active state.


The mammalian N-end rule pathway: new insights into its components and physiological roles.
The N-end rule pathway is a ubiquitin-dependent proteolytic system, in which destabilizing N-terminal residues of short-lived proteins function as an essential determinant of an N-terminal degradation signal (N-degron). An N-degron can be created from a pre-N-degron through specific N-terminal modifications, providing a means conditionally to destabilize otherwise stable polypeptides. The pathway has been found in all organisms examined, from prokaryotes to eukaryotes. Recent biochemical and proteomic studies identified many components of the mammalian N-end rule pathway, including a family of substrate recognition ubiquitin ligases and their substrates. The genetic dissection in animals and humans revealed its essential role in various vital physiological processes, ranging from cardiovascular development and meiosis to the pathogenesis of human genetic diseases. These discoveries have provided new insights into the components, functions and mechanics of this unique proteolytic system.


Exosomal circ-AHCY promotes glioblastoma cell growth via Wnt/β-catenin signaling pathway.
Glioblastoma (GBM) is the most aggressive brain tumor. Reportedly, circular RNAs (circRNAs) participate in regulation of the development and progression of diverse cancers, including GBM. Dysregulated circRNAs in GBM tissues were screened out from GEO database. The expression of candidate circRNAs in GBM cells was measured by qRT-PCR. Loss-of function assays, including colony formation assay, EdU assay, TUNEL assay, and flow cytometry analysis were conducted to determine the effects of circ-AHCY knockdown on GBM cell proliferation and apoptosis. Animal study was further used to prove the inhibitory effect of circ-AHCY silencing on GMB cell growth. Mechanistic experiments like luciferase reporter, RNA pull-down and RNA-binding protein immunoprecipitation (RIP) assays were performed to unveil the downstream molecular mechanism of circ-AHCY. Nanosight Nanoparticle Tracking Analysis (NTA) and PKH67 staining were applied to identify the existence of exosomes. Circ-AHCY was confirmed to be highly expressed in GBM cells. Circ-AHCY silencing suppressed GBM cell proliferation both in vitro and in vivo. Mechanistically, circ-AHCY activates Wnt/β-catenin signaling pathway by sequestering miR-1294 to up-regulate MYC which activated CTNNB1 transcription. It was also found that circ-AHCY recruited EIF4A3 to stabilize TCF4 mRNA. Enhanced levels of TCF4 and β-catenin contributed to the stability of TCF4/β-catenin complex. In turn, TCF4/β-catenin complex strengthened the transcriptional activity of circ-AHCY. Exosomal circ-AHCY derived from GBM cells induced abnormal proliferation of normal human astrocytes (NHAs). Exosomal circ-AHCY forms a positive feedback loop with Wnt/β-catenin signaling pathway to promote GBM cell growth.


Calcitriol in the Presence of Conditioned Media from Metastatic Breast Cancer Cells Enhances Ex Vivo Polarization of M2 Alternative Murine Bone Marrow-Derived Macrophages.
In this study, we differentiated murine bone marrow-derived macrophages (BMDMs) into M0, M1, and M2 in the presence or absence of calcitriol. Real-time PCR analysis of gene expression, FACS analysis of surface markers, and chemokine/cytokine production assays were performed. In addition, the effect of the conditioned media (CM) from murine breast cancer 4T1 (metastatic) and 67NR (non-metastatic) and Eph4-Ev (normal) cells with and without calcitriol on the polarization of M1/M2 cells was determined. We found that calcitriol enhanced the differentiation of M2 macrophages, which was manifested by increased expression of Cd206 and Spp1 mRNA and CD36, Arg, and CCL2 in M2 BMDMs and by decreased expression of Cd80 and Spp1 mRNA and IL-1, IL-6, OPN, and iNOS in M1 BMDMs. 4T1 CM showed a higher effect on the gene and protein expression in macrophages than 67NR and Eph4-Ev, with the greatest effect observed on M2 macrophages which increased their differentiation and properties characteristic of alternative macrophages. Moreover, M2 macrophages differentiated with calcitriol-stimulated migration of 4T1 and 67NR cells through fibronectin and collagen type IV, respectively. Overall, our results indicated that vitamin D supplementation may not always be beneficial, especially in relation to cancers causing excessive, pathological activation of the immune system.


Factors associated with high-risk drinking in older adults: Evidence from a national survey in Japan.
This study aimed to clarify the association between high-risk drinking and health outcomes (e.g. physical and psychosocial factors, including help-seeking behavior) among older Japanese adults. Data were obtained from the 2013 Comprehensive Survey of Living Conditions in Japan, which surveyed 295 367 households. We restricted analyses to 55 147 current drinkers. Multinomial logistic regression models were used to examine associations between alcohol consumption and various factors. Overall, 35.3% of participants were in the low-risk group, 13.1% were in the at-risk group and 1.0% were in the high-risk group. Those in the at-risk and high-risk groups were more likely to be younger, male, employed and tobacco users. They were also more likely to report sleeping ≥9 h per night, and they were more likely to report unhealthy eating habits. Being in the high-risk drinking group was significantly associated with being divorced (OR 1.51, 95% CI 1.11-2.05) and having poor mental health (OR 1.57, 95% CI 1.13-1.68). The high-risk drinking group was more likely to include those who endorsed experiencing challenges without seeking aid (OR 1.87, 95% CI 1.39-2.53) or felt no need to seek aid (OR 1.39, 95% CI 1.15-1.68). Being at high-risk for drinking is associated with poorer help-seeking behavior, suggesting a need for more aggressive outreach. Geriatr Gerontol Int 2019; 19: 1260-1267.


Steroid receptor coactivator-interacting protein (SIP) inhibits caspase-independent apoptosis by preventing apoptosis-inducing factor (AIF) from being released from mitochondria.
Apoptosis-inducing factor (AIF) is a caspase-independent death effector. Normally residing in the mitochondrial intermembrane space, AIF is released and translocated to the nucleus in response to proapoptotic stimuli. Nuclear AIF binds to DNA and induces chromatin condensation and DNA fragmentation, characteristics of apoptosis. Until now, it remained to be clarified how the mitochondrial-nuclear translocation of AIF is regulated. Here we report that steroid receptor coactivator-interacting protein (SIP) interacts directly with AIF in mitochondria and specifically inhibits caspase-independent and AIF-dependent apoptosis. Challenging cells with apoptotic stimuli leads to rapid degradation of SIP, and subsequently AIF is liberated from mitochondria and translocated to the nucleus to induce apoptosis. Together, our data demonstrate that SIP is a novel regulator in caspase-independent and AIF-mediated apoptosis.


Patients with Crohn's Disease Undergoing Abdominal Surgery: Clinical and Prognostic Evaluation Based on a Single-Center Cohort in China.
The incidence and prevalence of Crohn's disease (CD) are increasing in China, but there are few reports on the characteristics of patients requiring abdominal surgery. This study aimed to evaluate the clinical characteristics of these patients and the potential risk factors for postoperative complications and surgical recurrence. In this observational, retrospective single-center cohort analysis, patients with CD who had undergone at least one abdominal surgery at our center from 2007 to 2020 were included. Data were collected from a prospectively maintained database. Clinical factors were assessed by logistic regression models, Kaplan-Meier methods, and Cox proportional hazards regression models. The predictive accuracy of the nomogram was determined by a concordance index (C-index) and calibration curve and was validated using bootstrap resampling. In the 1639 patients, clinical characteristics were evaluated. In a multivariable logistic regression model, penetrating behavior (P = 0.002), emergency surgery (P = 0.010), and smoking status (P = 0.015) were significantly associated with an increased risk of postoperative septic complications. In contrast, staged surgery (P = 0.009) was inversely associated with postoperative complications. Upper gastrointestinal disease (P = 0.042), penetrating behavior (P = 0.027), emergency at initial surgery (P < 0.001) were significantly associated with an increased risk of surgical recurrence after the index surgery in our Cox regression model, whereas staged surgery (P = 0.036) was significantly associated with a decreased risk. The C-index of the nomogram for predicting recurrence was 0.744 (P = 0.015), and calibration curves showed good agreement between predictions of 3, 5, and 10 years of recurrence and actual observations. There are several disease- and surgery-associated risk factors of postoperative adverse outcomes in patients with CD undergoing abdominal surgery. This is important in optimizing the management of CD which has evolved into a global disease with rising prevalence in newly industrialized countries including China.


Detecting Myocardial Ischemia With 99mTechnetium-Tetrofosmin Myocardial Perfusion Imaging in Ischemic Stroke.
To assess the myocardial status in patients with stroke, employing myocardial perfusion imaging (MPI) with 99mTechnetium-tetrofosmin (99mTc-TF)-single-photon emission computed tomography (SPECT). Fifty-two patients with ischemic stroke were subjected to 99mTc-TF-SPECT MPI within 1 month after stroke occurrence. None of the patients had any history or symptoms of coronary artery disease or other heart disease. Myocardial perfusion imaging was evaluated visually using a 17-segment polar map. Myocardial ischemia (MIS) was defined as present when the summed stress score (SSS) was >4; MIS was defined as mild when SSS was 4 to 8, and moderate/severe with SSS ≥9. Patients with SSS >4 were compared to patients with SSS <4. Parameters such as age, body mass index, waist perimeter, smoking habits, and medical history (diabetes mellitus, dyslipidemia, etc) were evaluated according to MPI results. Myocardial ischemia was present in 32 (62%) of 52 patients with stroke. Among them, 20 (62%) of 32 patients had mild abnormalities and 12 (38%) of 32 had moderate/severe. The age and waist perimeter showed a tendency to relate to severe MIS when patients with SSS >9 were compared to patients with SSS <4. In MPI-positive patients, an age was to be association with SSS, with the oldest age exhibiting the highest SSS (P = .01). The association of age with SSS remained statistically significant in the multivariate analysis (P = .04). The study suggested that more than half of patients with stroke without a history of cardiac disease have MIS. Although most of them have mild MIS, we suggest a thorough cardiological evaluation in this group of patients for future prevention of severe myocardial outcome.