Confidence-based integrated reweighting model of task-difficulty explains location-based specificity in perceptual learning.
Perceptual learning is classically thought to be highly specific to the trained stimuli's retinal locations. However, recent research using a novel double-training paradigm has found dramatic transfer of perceptual learning to untrained locations. These results challenged existing models of perceptual learning and provoked intense debate in the field. Recently, Hung and Seitz (2014) showed that previously reported results could be reconciled by considering the details of the training procedure, in particular, whether it involves prolonged training at threshold using a single staircase procedure or multiple staircases. Here, we examine a hierarchical neural network model of the visual pathway, built upon previously proposed integrated reweighting models of perceptual learning, to understand how retinotopic transfer depends on the training procedure adopted. We propose that the transfer and specificity of learning between retinal locations can be explained by considering the task-difficulty and confidence during training. In our model, difficult tasks lead to higher learning of weights from early visual cortex to the decision unit, and thus to specificity, while easy tasks lead to higher learning of weights from later stages of the visual hierarchy and thus to more transfer. To model interindividual difference in task-difficulty, we relate task-difficulty to the confidence of subjects. We show that our confidence-based reweighting model can account for the results of Hung and Seitz (2014) and makes testable predictions.


Real‑time COVID-19 diagnosis from X-Ray images using deep CNN and extreme learning machines stabilized by chimp optimization algorithm.
Real-time detection of COVID-19 using radiological images has gained priority due to the increasing demand for fast diagnosis of COVID-19 cases. This paper introduces a novel two-phase approach for classifying chest X-ray images. Deep Learning (DL) methods fail to cover these aspects since training and fine-tuning the model's parameters consume much time. In this approach, the first phase comes to train a deep CNN working as a feature extractor, and the second phase comes to use Extreme Learning Machines (ELMs) for real-time detection. The main drawback of ELMs is to meet the need of a large number of hidden-layer nodes to gain a reliable and accurate detector in applying image processing since the detective performance remarkably depends on the setting of initial weights and biases. Therefore, this paper uses Chimp Optimization Algorithm (ChOA) to improve results and increase the reliability of the network while maintaining real-time capability. The designed detector is to be benchmarked on the COVID-Xray-5k and COVIDetectioNet datasets, and the results are verified by comparing it with the classic DCNN, Genetic Algorithm optimized ELM (GA-ELM), Cuckoo Search optimized ELM (CS-ELM), and Whale Optimization Algorithm optimized ELM (WOA-ELM). The proposed approach outperforms other comparative benchmarks with 98.25 % and 99.11 % as ultimate accuracy on the COVID-Xray-5k and COVIDetectioNet datasets, respectively, and it led relative error to reduce as the amount of 1.75 % and 1.01 % as compared to a convolutional CNN. More importantly, the time needed for training deep ChOA-ELM is only 0.9474 milliseconds, and the overall testing time for 3100 images is 2.937 s.


The roles of C-terminal loop residues of dimeric arginine kinase from sea cucumber Stichopus japonicus in catalysis, specificity and structure.
Arginine kinase (AK) catalyzes the reversible phosphorylation of arginine by MgATP to form a high-energy compound phosphoarginine (Parg) and MgADP in forward reaction in invertebrates. To detect the different catalytical mechanisms among Stichopus-AK (dimer) and Limulus-AK (monomer) and Torpedo creatine kinase (dimeric CK) and to reveal the structural role of the C-terminal domain loop (C-loop) of dimeric AK, six single-site mutants, E314D, E314Q, E314V, F315A, F315H and F315Y were constructed as well as two multi-site variants, S312R/F315H/V319E (formed by substituting the C-loop of monomeric AK for that of dimeric AK, termed the AAloop) and S312G/E314V/F315D/E317A/S318A/G321S (formed by substituting the C-loop of dimeric CK for that of dimeric AK, termed the ACloop). The AK activity of the three mutants at Glu(314) decreased significantly, from 60- to 500-fold. The ACloop showed only slight AK activity, unlike the same construction in Limulus-AK. In addition, all Phe(315) mutants including the AAloop which retained Glu(314) had modest AK activity (5-84% of the wild type). All the results above suggested that Glu(314) played a more significant role in catalysis in dimeric AK than in the monomer. In addition, ANS profiles indicated that the tolerance of the three Glu(314) mutants to denaturant decreased slightly compared with wild type AK. Though monomeric AK has a His residue at site 315, mutants F315H and the AAloop could not resist any perturbation of denaturant, and the mutants showed a Gibbs free energy of about 2.7kJ/mol lower than wild type AK. Therefore Phe(315) in dimeric AK has a different role from His(315) in monomeric AK. This might contribute to the stabilization of the native conformation, while His(315) in Limulus AK directly binded to the carboxylate of arginine. Taking all the results above together, we suggested a unique mechanism in dimeric AK, different from both monomeric AK and dimeric CK.


High-resolution melting genotyping of Enterococcus faecium based on multilocus sequence typing derived single nucleotide polymorphisms.
We have developed a single nucleotide polymorphism (SNP) nucleated high-resolution melting (HRM) technique to genotype Enterococcus faecium. Eight SNPs were derived from the E. faecium multilocus sequence typing (MLST) database and amplified fragments containing these SNPs were interrogated by HRM. We tested the HRM genotyping scheme on 85 E. faecium bloodstream isolates and compared the results with MLST, pulsed-field gel electrophoresis (PFGE) and an allele specific real-time PCR (AS kinetic PCR) SNP typing method. In silico analysis based on predicted HRM curves according to the G+C content of each fragment for all 567 sequence types (STs) in the MLST database together with empiric data from the 85 isolates demonstrated that HRM analysis resolves E. faecium into 231 "melting types" (MelTs) and provides a Simpson's Index of Diversity (D) of 0.991 with respect to MLST. This is a significant improvement on the AS kinetic PCR SNP typing scheme that resolves 61 SNP types with D of 0.95. The MelTs were concordant with the known ST of the isolates. For the 85 isolates, there were 13 PFGE patterns, 17 STs, 14 MelTs and eight SNP types. There was excellent concordance between PFGE, MLST and MelTs with Adjusted Rand Indices of PFGE to MelT 0.936 and ST to MelT 0.973. In conclusion, this HRM based method appears rapid and reproducible. The results are concordant with MLST and the MLST based population structure.


The acceptance and commitment therapy model in occupational rehabilitation of musculoskeletal and common mental disorders: a qualitative focus group study.
Aims: The aim of this study was to examine whether and how intended processes of behavioural change were reflected in participants' experiences after an inpatient occupational rehabilitation programme. The programme was transdiagnostic, lasted 3½ weeks and was based on the acceptance and commitment therapy model.Methods: Twenty-two participants (17 women and 5 men) took part in five qualitative focus group interviews after the programme. Analysis was data-driven, categorising participants' experiences using an initial explorative phenomenological approach. The emerging data-driven categories were re-contextualised within the theoretical framework of the therapy model.Results: The participants referred to experiences within all three intended domains of the model (openness, awareness, and engagement). Our results indicate that the transdiagnostic approach may have facilitated openness, while the attainment of flexible self-awareness was less evident. Participants expressed engagement and behavioural changes linked to personal values, but did not mention actions leading to imminent return to work.Conclusions: The results imply that for implementation in occupational rehabilitation, further development of this model is needed specifically regarding processes related to self-awareness and committed action towards work. These findings are relevant for the interpretation of results from randomised clinical trials on acceptance and commitment therapy in occupational rehabilitation.Implications for rehabilitationAcceptance and commitment therapy seems to be a feasible component in an occupational rehabilitation programme for persons with different diagnoses.A transdiagnostic approach mixing musculoskeletal pain disorders and common mental disorders in the same rehabilitation programme seems to facilitate the process of openness and acceptance.There is a need to further develop and operationalise some of the processes in Acceptance and commitment therapy to accommodate the approach to the occupational rehabilitation context.


Seagrass habitat metabolism increases short-term extremes and long-term offset of CO2 under future ocean acidification.
The role of rising atmospheric CO2 in modulating estuarine carbonate system dynamics remains poorly characterized, likely due to myriad processes driving the complex chemistry in these habitats. We reconstructed the full carbonate system of an estuarine seagrass habitat for a summer period of 2.5 months utilizing a combination of time-series observations and mechanistic modeling, and quantified the roles of aerobic metabolism, mixing, and gas exchange in the observed dynamics. The anthropogenic CO2 burden in the habitat was estimated for the years 1765-2100 to quantify changes in observed high-frequency carbonate chemistry dynamics. The addition of anthropogenic CO2 alters the thermodynamic buffer factors (e.g., the Revelle factor) of the carbonate system, decreasing the seagrass habitat's ability to buffer natural carbonate system fluctuations. As a result, the most harmful carbonate system indices for many estuarine organisms [minimum pHT, minimum Ωarag, and maximum pCO2(s.w.)] change up to 1.8×, 2.3×, and 1.5× more rapidly than the medians for each parameter, respectively. In this system, the relative benefits of the seagrass habitat in locally mitigating ocean acidification increase with the higher atmospheric CO2 levels predicted toward 2100. Presently, however, these mitigating effects are mixed due to intense diel cycling of CO2 driven by aerobic metabolism. This study provides estimates of how high-frequency pHT, Ωarag, and pCO2(s.w.) dynamics are altered by rising atmospheric CO2 in an estuarine habitat, and highlights nonlinear responses of coastal carbonate parameters to ocean acidification relevant for water quality management.


Updates on the web-based VIOLIN vaccine database and analysis system.
The integrative Vaccine Investigation and Online Information Network (VIOLIN) vaccine research database and analysis system (http://www.violinet.org) curates, stores, analyses and integrates various vaccine-associated research data. Since its first publication in NAR in 2008, significant updates have been made. Starting from 211 vaccines annotated at the end of 2007, VIOLIN now includes over 3240 vaccines for 192 infectious diseases and eight noninfectious diseases (e.g. cancers and allergies). Under the umbrella of VIOLIN, >10 relatively independent programs are developed. For example, Protegen stores over 800 protective antigens experimentally proven valid for vaccine development. VirmugenDB annotated over 200 'virmugens', a term coined by us to represent those virulence factor genes that can be mutated to generate successful live attenuated vaccines. Specific patterns were identified from the genes collected in Protegen and VirmugenDB. VIOLIN also includes Vaxign, the first web-based vaccine candidate prediction program based on reverse vaccinology. VIOLIN collects and analyzes different vaccine components including vaccine adjuvants (Vaxjo) and DNA vaccine plasmids (DNAVaxDB). VIOLIN includes licensed human vaccines (Huvax) and veterinary vaccines (Vevax). The Vaccine Ontology is applied to standardize and integrate various data in VIOLIN. VIOLIN also hosts the Ontology of Vaccine Adverse Events (OVAE) that logically represents adverse events associated with licensed human vaccines.


The gene for the Alzheimer-associated beta-amyloid-binding protein (ERAB) is differentially expressed in the testicular Leydig cells of the azoospermic by w/w(v) mouse.
In order to discover possible new testicular paracrine factors involved in the establishment of spermatogenesis, a modified differential display reverse transcription, polymerase chain reaction (DDRT-PCR) procedure was used to detect gene transcripts preferentially expressed in the testes of the azoospermic w/w(v) mutant mouse. One of the differentially expressed gene products showed partial similarity to members of the short-chain alcohol dehydrogenase family of enzymes. This cDNA fragment was used to obtain the full-length mouse cDNA sequence, which initially showed moderate similarity to a 20beta-steroid dehydrogenase from lower organisms, and later shown to have >85% similarity to a novel endoplasmic-reticulum-associated-binding protein (ERAB) from the human brain, implicated in Alzheimer's disease. A recently cloned bovine sequence also of high similarity suggests that this molecule might also represent an isozyme of 3-hydroxyacyl-CoA dehydrogenase. Using the mouse cDNA as probe, northern hybridization showed enrichment of the transcript to the testicular Leydig cells, and showed a specific approximately 20-fold enrichment in the azoospermic mouse testis. The level of the testicular ERAB transcript does not seem to change through puberty, suggesting that a lack of germ cells alone is not responsible for the up-regulation in the w/w(v) testis. Using the three-dimensional coordinates of the published 20beta-hydroxysteroid dehydrogenase structure as template, it was additionally possible to construct a molecular model of the novel protein which showed it to have a very similar structure to this enzyme, including the substrate-binding domain.


Analysis of proteomic profiles and functional properties of human peripheral blood myeloid dendritic cells, monocyte-derived dendritic cells and the dendritic cell-like KG-1 cells reveals distinct characteristics.
Dendritic cells (DCs) are specialized antigen presenting cells that play a pivotal role in bridging innate and adaptive immune responses. Given the scarcity of peripheral blood myeloid dendritic cells (mDCs) investigators have used different model systems for studying DC biology. Monocyte-derived dendritic cells (moDCs) and KG-1 cells are routinely used as mDC models, but a thorough comparison of these cells has not yet been carried out, particularly in relation to their proteomes. We therefore sought to run a comparative study of the proteomes and functional properties of these cells. Despite general similarities between mDCs and the model systems, moDCs and KG-1 cells, our findings identified some significant differences in the proteomes of these cells, and the findings were confirmed by ELISA detection of a selection of proteins. This was particularly noticeable with proteins involved in cell growth and maintenance (for example, fibrinogen gamma chain (FGG) and ubiquinol cytochrome c) and cell-cell interaction and integrity (for example, fascin and actin). We then examined the surface phenotype, cytokine profile, endocytic and T-cell-activation ability of these cells in support of the proteomic data, and obtained confirmatory evidence for differences in the maturation status and functional attributes between mDCs and the two DC models. We have identified important proteomic and functional differences between mDCs and two DC model systems. These differences could have major functional implications, particularly in relation to DC-T cell interactions, the so-called immunological synapse, and, therefore, need to be considered when interpreting data obtained from model DC systems.


Orbital angular momentum mode groups multiplexing transmission over 2.6-km conventional multi-mode fiber.
Twisted light carrying orbital angular momentum (OAM) is a special kind of structured light that has a helical phase front, a phase singularity, and a doughnut intensity profile. Beyond widespread developments in manipulation, microscopy, metrology, astronomy, nonlinear and quantum optics, OAM-carrying twisted light has seen emerging application of optical communications in free space and specially designed fibers. Instead of specialty fibers, here we show the direct use of a conventional graded-index multi-mode fiber (MMF) for OAM communications. By exploiting fiber-compatible mode exciting and filtering elements, we excite the first four OAM mode groups in an MMF. We demonstrate 2.6-km MMF transmission using four data-carrying OAM mode groups (OAM0,1, OAM+1,1/OAM-1,1, OAM+2,1, OAM+3,1). Moreover, we demonstrate two data-carrying OAM mode groups multiplexing transmission over the 2.6-km MMF with low-level crosstalk free of multiple-input multiple-output digital signal processing (MIMO-DSP). The demonstrations may open up new perspectives to fiber-based OAM communication/non-communication applications using already existing conventional fibers.


Linkage disequilibrium and effective population size when generations overlap.
Estimates of effective population size are critical for species of conservation concern. Genetic datasets can be used to provide robust estimates of this important parameter. However, the methods used to obtain these estimates assume that generations are discrete. We used simulated data to assess the influences of overlapping generations on the estimates of effective size provided by the linkage disequilibrium (LD) method. Our simulations focus on two factors: the degree of reproductive skew exhibited by the focal species and the generation time, without considering sample size or the level of polymorphism at marker loci. In situations where a majority of reproduction is achieved by a small fraction of the population, the effective number of breeders can be much smaller than the per-generation effective population size. The LD in samples of newborns can provide estimates of the former size, while our results indicate that the latter size is best estimated using random samples of reproductively mature adults. Using samples of adults, the downwards bias was less than approximately 15% across our simulated life histories. As noted in previous assessments, precision of the estimate depends on the magnitude of effective size itself, with greater precision achieved for small populations.


High c-Myb Expression Associates with Good Prognosis in Colorectal Carcinoma.
Colorectal cancer (CRC) represents a serious challenge for oncologists due to high incidence and large heterogeneity. Prognostic factors are needed to stratify patients according to risk of disease progression. In this study, we report that high expression of c-Myb protein, determined by immunohistochemistry (IHC), associates with better overall and disease-free survival (OS, DFS) in a cohort of 103 patients. Although MYB has been previously considered to act as oncogene in CRC, our further analysis of datasets deposited in PrognoScan and SurvExpress databases confirmed that high MYB expression largely associates with good prognosis in CRC. As therapies targeting c-Myb have been developed and tested in preclinical studies, we believe that further studies are needed for detailed understanding of c-Myb function in CRC, before the c-Myb-targeted therapy enters clinical trials.


Death-and-culling rates of calves and associated economic losses in the first month of life on dairy farms in eastern Hokkaido, Japan.
The objective of this study was to analyze death and culling (DC) of calves during the first month of life and associated economic losses on dairy farms in eastern Hokkaido, Japan. The DC in the first month of life of 4411 Holstein and Wagyu crossbred calves born in the year 2019-2020 on 39 dairy farms milking Holsteins was investigated. Based on a target DC rate of 6.75%, farms were classified into two groups, those with high DC rates (HDC, 11.68%, n = 18) and those with low DC rates (LDC, 2.67%, n = 21), and analyzed for DC factors (breed, sex, parity of dams, and housing type of dams) and diseases causing DC, their loss estimates, and replenishment of DC calves (birth rate, purchase of heifers, and housing type of dams). Comparisons between groups were made using the Kaplan-Meier method, the Mann-Whitney U test, and chi-square test. The DC rate of Holsteins was significantly higher (P < 0.001) for HDC farms than for LDC ones. But, the DC rate of Wagyu crossbreds was not different between the groups, which suggested hybrid vigor. The DC rates for digestive diseases were significantly higher (P < 0.01) in the HDC farms, except for congenital diseases and deaths of unknown cause. The overall loss estimates of DC per calf-month for all farms was 8892, JPY/calf-months, 14,726 for HDC farms, and 4065 for LDC farms. The loss estimates of items with significant differences in DC rates were significantly higher (P < 0.05) in the HDC farms, with higher loss estimates for Holsteins and digestive diseases being the most common characteristics of HDC farms. In binomial logistic regression analysis with "HDC farms or not" as the response variable and replenishment of DC calves as the explanatory variable, HDC was significantly more likely (OR: 1.10, P < 0.05) on farms with a higher birth proportion of Holsteins, and HDC farms supplemented the DC calves by increasing the birth proportion of Holsteins.


Effect of Titanium Miniplate Fixation on Hinge Fracture and Hinge Fracture Displacement Following Cervical Open-Door Laminoplasty.
Cervical spondylotic myelopathy is a neuromotor disorder responsible for functional limitations and decreased daily activities. Expansive open-door laminoplasty is the widely accepted procedure for the treatment of multilevel cervical spondylotic myelopathy. Among the various fixation procedures to secure the open lamina, miniplate fixation provides better clinical and radiological outcomes. However, the immediate effects on hinge fracture and hinge fracture displacement following miniplate fixation have not been proven until now. The purpose of our study was to elucidate the impact of cervical open-door angle on the status of spinal cord expansion and hinge fracture, hinge fracture displacement, and the role of implants used during surgery. For this retrospective study, 122 patients who had undergone surgery from September 2016 to November 2017 with preoperative and postoperative radiographs were enrolled. Clinical and radiological outcomes were assessed before and after surgery. There were no significant differences in demographics, surgery time, blood loss, medical comorbidities, or perioperative and postoperative complications between 2 groups. The recovery rate and Nurick score before and at the follow-up show no statistical significance between the 2 groups, P value > .05 (P = .672) and P > .05 (P = .553), respectively. The statistical analysis shows that the mean hinge fracture in the miniplate group with a cervical open angle >30° was 2.42 ± 1.68 and with a <30° open angle, 0.05 ± 0.23; whereas, in the anchor group the mean hinge fracture in >30° cervical open angle was 2.227 ± 2.50 and in <30° was 0.409 ± 0.503. The results revealed statistical significance between 2 implant groups, P = .024 in the aspect of hinge fracture displacement and implant used. Laminoplasty by titanium miniplate fixation holds the laminae securely, prevents hinge fracture displacement, and promotes spinal cord expansion better than suture anchor fixation.


Treating Agricultural Runoff with a Mobile Carbon Filtration Unit.
Several classes of pesticides have been shown to impair water quality in California, including organophosphates, pyrethroids and neonicotinoids. Vegetative treatment systems (VTS) can reduce pesticide loads and associated toxicity in agricultural runoff, but many water-soluble pesticides such as neonicotinoids are not effectively treated by VTS, and VTS installation is not always an option for growers required to remove non-crop vegetation for food safety concerns. Recent studies have shown that biochar filtration can be used to remove soluble contaminants, especially when coupled with other VTS components. We evaluated a mobile carbon filter system consisting of a trailer-mounted tank containing approximately 600L (~ 180 kg) of biochar. Input water from a 437-hectare agricultural drainage was pre-filtered and treated with biochar during two multi-week study periods. Laboratory toxicity tests and chemical and nutrient analyses were conducted on input and output water. Pesticide concentrations were initially reduced by greater than 99%. Treatment efficacy declined linearly and was expected to remain at least 50% effective for up to 34 weeks. Toxicity was assessed with Ceriodaphnia dubia, Hyalella azteca and Chironomus dilutus. Significant input toxicity was reduced to non-toxic levels in 6 of 16 samples. Some input concentrations of the neonicotinoid imidacloprid and the pyrethroid cypermethrin exceeded organism-specific toxicity thresholds and benchmarks, but the overall causes of toxicity were complex mixtures of agricultural chemicals. Nutrients were not reduced by the biochar. Results demonstrate the utility of biochar in treating agricultural runoff and provide measures of the longevity of biochar under field conditions.


A novel and potent poly(ADP-ribose) polymerase-1 inhibitor, FR247304 (5-chloro-2-[3-(4-phenyl-3,6-dihydro-1(2H)-pyridinyl)propyl]-4(3H)-quinazolinone), attenuates neuronal damage in in vitro and in vivo models of cerebral ischemia.
The activation of poly(ADP-ribose) polymerase-1 (PARP-1) after exposure to nitric oxide or oxygen-free radicals can lead to cell injury via severe, irreversible depletion of NAD. Genetic deletion or pharmacological inhibition of PARP-1 attenuates brain injury after focal ischemia and neurotoxicity in several neurodegenerative models in animals. FR247304 (5-chloro-2-[3-(4-phenyl-3,6-dihydro-1(2H)-pyridinyl)propyl]-4(3H)-quinazolinone) is a novel PARP-1 inhibitor that has recently been identified through structure-based drug design. In an enzyme kinetic analysis, FR247304 exhibits potent and competitive inhibition of PARP-1 activity, with a K(i) value of 35 nM. Here, we show that prevention of PARP activation by FR247304 treatment protects against both reactive oxygen species-induced PC12 cell injury in vitro and ischemic brain injury in vivo. In cell death model, treatment with FR247304 (10(-8)-10(-5) M) significantly reduced NAD depletion by PARP-1 inhibition and attenuated cell death after hydrogen peroxide (100 microM) exposure. After 90 min of middle cerebral artery occlusion in rats, poly(ADP-ribosy)lation and NAD depletion were markedly increased in the cortex and striatum from 1 h after reperfusion. The increased poly(ADP-ribose) immunoreactivity and NAD depletion were attenuated by FR247304 (32 mg/kg i.p.) treatment, and FR247304 significantly decreased ischemic brain damage measured at 24 h after reperfusion. Whereas other PARP inhibitors such as 3-aminobenzamide and PJ34 [N-(6-oxo-5,6-dihydro-phenanthridin-2-yl)-N,N-dimethylactamide] showed similar neuroprotective actions, they were less potent in in vitro assays and less efficacious in an in vivo model compared with FR247304. These results indicate that the novel PARP-1 inhibitor FR247304 exerts its neuroprotective efficacy in in vitro and in vivo experimental models of cerebral ischemia via potent PARP-1 inhibition and also suggest that FR247304 or its derivatives could be attractive therapeutic candidates for stroke and neurodegenerative disease.


Transcatheter Aortic Valve Implantation in Patients with Connective Tissue Disease.
There is still no consensus on the treatment of patients with connective tissue disease (CTD) with severe symptomatic aortic stenosis (AS). The aim of this study was to evaluate the feasibility and safety of transcatheter aortic valve implantation (TAVI) in patients with CTD. Five hundred and fifty consecutive symptomatic severe AS patients who underwent TAVI between 2011 and 2019 were included in this retrospective study, of whom 14 had CTD. Follow-up was performed 30 days, 6 months, and 1 year after the procedure. Of the 14 (2.5%) patients who had CTD, most had rheumatoid arthritis (n = 10), followed by lupus erythematosus (n = 2), scleroderma (n = 1) and mixed (n = 1) CTD. The mean age was 77.6 ± 7.9 years, and there was no statistical difference between the CTD and no-CTD groups. In addition, significantly more of the CTD patients (85.7%) were female compared to the no-CTD group (p = 0.018). None of the patients in the CTD group had acute kidney injury, stroke, major bleeding, or pericardial effusion. However, significantly more patients in the CTD group (n = 4) needed permanent pacemaker implantation than in the no-CTD group (p = 0.008). There were no significant differences between the two groups in terms of mean discharge time (CTD 4.6 ± 2.0, no-CTD 4.5 ± 2.3 days, p = 0.926) and in-hospital mortality [CTD 1 (7.1%), no-CTD 21 (3.9%); p = 0.542]. In this study, we presented the results of TAVI in patients with and without CTD. The TAVI procedure had similar mid-term outcomes in the two groups, and the CTD group had numerically lower rates of major complications at the cost of a higher incidence of pacemaker implantation.


Signal scavenging for passive monitoring in eldercare technology.
Signal scavenging is analogous to energy scavenging: seemingly ubiquitous energy in the environs provides the signal for usage as a personnel sensor. Such energy can be used to detect motion, and most importantly falls. Stray signals can be detected in aluminum foil as voltage differences between touched foil (say by hand) compared to that untouched. Spectrum analysis shows the stray electromagnetic noise signal consists substantially of 60 Hz and its harmonics. Also the signal intensity for both touched and untouched monotonically increases with foil area. While personnel monitors find utility in many areas including security, personnel control and activity detection, we believe these putative sensors to be useful in inobtrusive monitoring of elders to provide them with increased independence at a critical time in their lives.


Combining diverse evidence for gene recognition in completely sequenced bacterial genomes.
Analysis of a newly sequenced bacterial genome starts with identification of protein-coding genes. Functional assignment of proteins requires the exact knowledge of protein N-termini. We present a new program ORPHEUS that identifies candidate genes and accurately predicts gene starts. The analysis starts with a database similarity search and identification of reliable gene fragments. The latter are used to derive statistical characteristics of protein-coding regions and ribosome-binding sites and to predict the complete set of genes in the analyzed genome. In a test on Bacillus subtilis and Escherichia coli genomes, the program correctly identified 93.3% (resp. 96.3%) of experimentally annotated genes longer than 100 codons described in the PIR-International database, and for these genes 96.3% (83.9%) of starts were predicted exactly. Furthermore, 98.9% (99.1%) of genes longer than 100 codons annotated in GenBank were found, and 92.9% (75.7%) of predicted starts coincided with the feature table description. Finally, for the complete gene complements of B.subtilis and E.coli , including genes shorter than 100 codons, gene prediction accuracy was 88.9 and 87.1%, respectively, with 94.2 and 76.7% starts coinciding with the existing annotation.


Piezo1 transforms mechanical stress into pro senescence signals and promotes osteoarthritis severity.
Osteoarthritis (OA) is a prevalent disease among elderly people and is often characterized by chronic joint pain and dysfunction. Recently, growing evidence of chondrocyte senescence in the pathogenesis of OA has been found, and targeting senescence has started to be recognized as a therapeutic approach for OA. Piezo1, a mechanosensitive Ca2+ channel, has been reported to be harmful in sensing abnormal mechanical overloading and leading to chondrocyte apoptosis. However, whether Piezo1 can transform mechanical signals into senescence signals has rarely been reported. In this study, we found that severe OA cartilage expressed more Piezo1 and the senescence markers p16 and p21. 24 h of periodic mechanical stress induced chondrocyte senescence in vitro. In addition, we demonstrated the pivotal role of Piezo1 in OA chondrocyte senescence induced by mechanical stress. Piezo1 sensed mechanical stress and promoted chondrocyte senescence via its Ca2+ channel ability. Moreover, Piezo1 promoted SASP factors production under mechanical stress, particularly in IL-6 and IL-1β. p38MAPK and NF-κB activation were two key pathways that responded to Piezo1 activation and promoted IL-6 and IL-1β production, respectively. Collectively, our study revealed a connection between abnormal mechanical stress and chondrocyte senescence, which was mediated by Piezo1.


[Analysis of immunophenotype and leukemia associated immunophenotype in 610 patients with acute myeloid leukemia].
To analyze the immunophenotype and leukemia associated immunophenotype (LAIP) of leukemia cells from patients with acute myeloid leukemia (AML) in minimal residual disease (MRD) detection. Four-color multiparametric flow cytometry (FCM) with CD45/SSC gating was used to determine the expression of the following antibodies of CD7, CD117, CD33, CD34, CD10, CD19, CD56, CD38, CD13, CD14, CD64, CD9, CD16, CD2, CD5, CD11b, CD123, HLA-DR in 610 AML patients and 20 normal bone marrow (NBM) samples. The mean percentages of CD34+ and CD117+ side scatter low (SSC(low)) cells in NBM mononuclear cells (BMMNCs) were (0.35 +/- 0.15)% and (0.76 +/- 0.31)%, respectively. The majority (84% -95%) of CD34+ SSC(low) cells co-expressed CD13, CD33, CD38, CD117 and HLA-DR. 33% and 20% of CD34+ SSC(low) cells were CD15+ and CD9, respectively. Only a small proportion (< 10%) of CD34 SSC(low) cell co-expressed CD11b, CD56, CD19 and CD64, while co-expressed CD7 was 12%. In CD117 SSC(low) cells, the relative proportions of CD19+, CD11b+, CD56+ and CD7+ were less than 10%, while CD9+ was 14%. CD38+ and HLA-DR+ were 87% and 91%, respectively. The expressions of CD15, CD34, CD13 and CD33 on CD117 SSC(low) cells were between 30% and 50%. In AML patients, most cases were CD117+ (95.08%), CD38+ (94.74%), CD9+ (84.93%), CD33+ (84.60%), HLA-DR+ (77.23%) and CD13 (75.25%). The proportions of CD64+ and CD34+ were 64.41% and 59.51%, and that of CD15 and CD11b+ were 43.06% and 22.07%, respectively. 86.39% of AML patients were found to have at least one LAIP, the highest incidence being in AML-M1 and M3 subtypes and the lowest in AML-M4Eo subtype. The cross-lineage antigen and asynchronous antigen expression were the most frequent aberrant phenotypes. CD7, CD19 and CD56 expressing on CD34+ cells were major cross-lineage antigen. For asynchronous antigen expression, CD34+ CD64+, CD117+ CD11b+ , CD34+ CD38(-/dim) and CD34+ HLA-DR(-/dim) were seldom expressed on normal BMMNCs (about 0.01%), and the logarithm difference between AML and NBM was larger than 3.0, being the more sensitive LAIP. MRD detection by multiparameter flow cytometry can be applied to more than 80% of AML patients.


Exploring issues of participation among adolescents with cerebral palsy: what's important to them?
The purpose of this cross-sectional study was to determine what participation issues are important to adolescents with cerebral palsy (CP). Two hundred and three adolescents with CP (mean age 16.0 ± 1.8 years) were assessed using the Canadian Occupational Performance Measure (COPM). This was done through semistructured interviews by trained physical and occupational therapists. Adolescents responded either directly (n = 144) or through a parent or a caregiver (n = 59) if they were unable to communicate. Issues were extracted from completed questionnaires and coded under three COPM categories (self-care, productivity, and leisure) and 16 subcategories. There was no association between the total number of issues identified and gender (p = .99), age (p = .88), type of respondent (adolescents versus parent) (p = .27), Gross Motor Function Classification System (GMFCS) level (p = .93), or 66-item Gross Motor Function Measure (GMFM-66) score (p = .45). The issues identified most frequently were related to active leisure (identified by 57% of participants), mobility (55%), school (48%), and socialization (44%). Interventions aimed at improving participation among adolescents with physical disabilities, such as CP, should be directed towards these four key areas. Health care professionals should also recognize and consider the interaction of person and environment when addressing issues related to participation.


Multi-omics analyses with stool-type stratification in patient cohorts and Blautia identification as a potential bacterial modulator in T2DM.
Heterogeneity in host and gut microbiota hampers microbial precision intervention of type 2 diabetes mellitus (T2DM). Here, we investigate novel features for patient-stratification and bacterial modulators for intervention, using cross-sectional patient cohorts and animal experiments. We collected stool/blood/urine samples from 103 recent-onset T2DM patients and 25 healthy controls (HCs), performed gut microbial composition/metabolite profiling, and combined it with host-transcriptome/metabolome/cytokines and clinical data. Stool-type (dry/loose-stool), a feature of stool-microenvironment recently explored in microbiome studies, was used for T2DM patientstratification as it explained most of the variation in multi-omics dataset among all clinical parameters in our covariate analysis. T2DM with dry-stool (DM-DS) and loose-stool (DM-LS) were clearly differentiated from HC and each other by LightGBM-models, optimal among multiple machine-learning models. Compared to DM-DS, DM-LS exhibited discordant gut microbial taxonomic and functional profiles, severe host metabolic disorder, and excessive insulin secretion. Further cross-measurement-association-analysis linked the differential microbial profiles, in particular Blautia abundances, to T2DM phenotypes in our stratified multi-omics dataset. Notably, oral supplementation of Blautia to T2DM mice induced inhibitory effects on lipid accumulation, weight gain, and blood-glucose elevation with simultaneous modulation of gut bacterial composition, revealing the therapeutic potential of Blautia. Our study highlights the clinical implications of stool-microenvironment stratification and Blautia supplementation in T2DM, offering promising prospects for microbial precision treatment of metabolic diseases.


The Keys to Success in Doctoral Studies: A Preimmersion Course.
This article will review an innovative on-line preimmersion course for a hybrid doctor of nursing practice (DNP) program and a traditional face-to-face doctor of philosophy nursing program. The doctoral candidates include both postbaccalaureate and postmaster's students. The authors of the preimmersion course developed and initiated the course in order to address various issues that have surfaced in discussions between students and faculty. Examples of common themes identified include writing skills, statistics, life-work-school balance, and navigating instructional technology. Doctoral studies may pose challenges to students studying nursing, in regard to academic rigor and experiencing on-line education for the first time, especially for students who have been out of school for an extended amount of time or are not accustomed to a nontraditional classroom; thus, having a preimmersion course established may facilitate a smooth transition to rigorous academic studies in a hybrid program. The course, which was developed and delivered through Blackboard, a learning management system, includes the following 9 preimmersion modules: academic strategies (learning styles, creating an effective PowerPoint presentation), library support (introduction to the university library, literature review tutorial, and citation styles), mindfulness, wellness, statistics essentials, writing express, DNP capstone, netiquette, and DNP/doctor of philosophy mentorship. Each module consists of various tools that may promote student success in specific courses and the programs in general. The purpose of designing the preimmersion course is to decrease attrition rates and increase success of the students. While the majority of students have succeeded in their coursework and been graduated from the program, the authors of this article found that many students struggled with the work, life, and school balance. Future work will include the evaluation of results from graduate students enrolled in the program.


Novel structural determinants of the human neutrophil chemotactic activity of leukotriene B.
A specific 5(S),12(R)-dihydroxy-eicosa-6,8,10(trans/trans/cis), 14(cis)-tetraenoic acid, designated leukotriene B, is generated by the lipoxygenation and subsequent enzymatic hydration of arachidonic acid in a variety of leukocytes. Leukotriene B elicits a maximal human neutrophil chemotactic response in vitro which is similar in magnitude to those evoked by the chemotactic fragment of the fifth component of complement, C5a, synthetic formyl-methionyl peptides, and 5-hydroxy-eicosatetraenoic acid (5-HETE). The neutrophil chemotactic potency of purified leukotriene B, assessed by the 50% effective concentration of 6 x 10(-9) M, is equivalent to that of C5a, but is up to 100-fold greater than that of 5-HETE and of other natural di-HETE isomers. 5(S),12(R)-di-hydroxy-eicosa-6,8,10(all-trans),14(cis)-tetraenoic acid, which differs from leukotriene B only in having a trans-double bond in place of a cis-double bond in the triene portion of the molecule, and acetyl-leukotriene B are significantly less potent neutrophil chemotactic factors than leukotriene B, which indicates that both the conjugated double bonds and the free hydroxyl-group(s) are functionally critical determinants. The capacity of acetyl-leukotriene B to inhibit competitively and selectively the human neutrophil chemotactic response to equimolar concentrations of leukotriene B suggests the existence of a specific subset of receptors for this potent lipid mediator.


ΩqPCR measures telomere length from single-cells in base pair units.
ΩqPCR determines absolute telomere length in kb units from single cells. Accuracy and precision of ΩqPCR were assessed using 800 bp and 1600 bp synthetic telomeres inserted into plasmids, which were measured to be 819 ± 19.6 and 1590 ± 42.3 bp, respectively. This is the first telomere length measuring method verified in this way. The approach uses Ω-probes, a DNA strand containing sequence information that enables: (i) hybridization with the telomere via the 3' and 5' ends that become opposed; (ii) ligation of the hybridized probes to circularize the Ω-probes and (iii) circularized-dependent qPCR due to sequence information for a forward primer, and for a reverse primer binding site, and qPCR hydrolysis probe binding. Read through of the polymerase during qPCR occurs only in circularized Ω-probes, which quantifies their number that is directly proportional to telomere length. When used in concert with information about the cell cycle stage from a single-copy gene, and ploidy, the MTL of single cells measured by ΩqPCR was consistent with that obtained from large sample sizes by TRF.


Methylsulfonylmethane exhibits bacteriostatic inhibition of Escherichia coli, and Salmonella enterica Kinshasa, in vitro.
To evaluate antibacterial properties of methylsulfonylmethane (MSM) on Escherichia coli (MDRE21) and Salmonella enterica serovar Kinshasa (SK132). Bacterial proliferation analysis was measured spectrophotometrically during log phase growth with 0, 3, 5, 7, 10, 12 and 16% MSM. To assess the mechanism of inhibition, cultures were grown overnight with 0-16% MSM and enumerated on unmedicated brain-heart infusion agar (BHIA) or BHIA with 0-16% MSM. The long-term viability studies were done to evaluate the impact of 10% MSM. Absorbance data indicated a dose-dependent inhibition from 0 to 16% MSM. There was no growth of MDRE21 or SK132 on BHIA in 10-16% MSM. Both strains enumerated on unmedicated BHIA from overnight cultures with 10-16% MSM were able to partially recover. Recovery after MSM removal may be indicative of a bacteriostatic mechanism of inhibition. The long-term viability studies illustrated that neither MDRE21 nor SK132 could be rescued from 10% MSM after 5 or 6 days respectively. Methylsulfonylmethane antibacterial activity may prove useful during pre or postharvest food safety as a disinfectant. The primary benefit being, its clinical safety to humans. Comparisons to other disinfectants would also need to be done to determine if MSM was superior to those already on the market and would be cost effective.


The Impact of Proactive Gastrostomy Tube Placement on Treatment-related Outcomes in Young Children With High-grade Central Nervous System Tumors.
Young children undergoing treatment with intensive chemotherapy for high-grade central nervous system (CNS) tumors are at risk for malnutrition, yet no guidelines exist for the placement of enteral tubes. Prior studies evaluated the impact of proactive gastrostomy tube (GT) placement with a narrow scope of outcomes, such as weight. To examine the impact of proactive GT on comprehensive treatment outcomes, we performed a single-center, retrospective study of children younger than 60 months of age with high-grade CNS tumors treated per CCG99703 or ACNS0334 between 2015 and 2022. Of 26 patients included, 9 (35%) underwent proactive GT, 8 (30%) had rescue GT, and 9 (35%) had a nasogastric tube (NGT). Clinically significant weight loss occurred in 47% of patients with NGT during induction compared with 22% with proactive GT ( P = 0.274); however, between cohorts, there was no significant difference in antibiotic or parenteral nutrition utilization, weight loss at therapy completion, and duration of hospitalization. Therefore, proactive GT placement was modestly effective at preventing significant weight loss during induction, however, there was no clear benefit for hospitalization duration, antibiotic, or parental nutrition requirements compared with NGT. We recommend an individualized approach to GT placement for young children with CNS malignancies undergoing intensive chemotherapy.


The Sulu-Sulawesi Sea: environmental and socioeconomic status, future prognosis and ameliorative policy options.
The Sulu-Sulawesi Sea, with neighboring Indonesian Seas and South China Sea, lies at the center of the world's tropical marine biodiversity. Encircled by 3 populous, developing nations, the Philippines, Indonesia and Malaysia, the Sea and its adjacent coastal and terrestrial ecosystems, supports ca. 33 million people, most with subsistence livelihoods heavily reliant on its renewable natural resources. These resources are being impacted severely by rapid population growth (> 2% yr-1, with expected doubling by 2035) and widespread poverty, coupled with increasing international market demand and rapid technological changes, compounded by inefficiencies in governance and a lack of awareness and/or acceptance of some laws among local populations, particularly in parts of the Philippines and Indonesia. These key root causes all contribute to illegal practices and corruption, and are resulting in severe resource depletion and degradation of water catchments, river, lacustrine, estuarine, coastal, and marine ecosystems. The Sulu-Sulawesi Sea forms a major geopolitical focus, with porous borders, transmigration, separatist movements, piracy, and illegal fishing all contributing to environmental degradation, human suffering and political instability, and inhibiting strong trilateral support for interventions. This review analyzes these multifarious environmental and socioeconomic impacts and their root causes, provides a future prognosis of status by 2020, and recommends policy options aimed at amelioration through sustainable management and development.


Lutetium-177 prostate-specific membrane antigen (PSMA) theranostics: practical nuances and intricacies.
Theranostic principles utilize a molecular biomarker specific for a tumor target, initially for imaging to assess target expression and, if deemed suitable, for targeted therapy. This presents an exciting opportunity for a highly personalized treatment strategy in the era of precision medicine. Prostate-specific membrane antigen (PSMA) theranostics has attracted increasing attention as a promising targeted treatment in metastatic prostate cancer (PC). 177Lu-DOTA-PSMA-617 (177Lu-PSMA-617) is a PSMA-targeted small molecule with favorable properties and is the most extensively investigated PSMA radioligand for radionuclide therapy (RNT) in PC. Since 2014 multiple retrospective studies and more recently a phase II prospective study demonstrated safety and impressive efficacy of 177Lu-PSMA RNT. The evidence generated by these trials led to two currently underway randomized trials in metastatic castrate-resistant PC: TheraP (NCT03392428) and VISION (NCT03511664). While we wait for these pivotal trials to read out, nuclear medicine physicians, medical oncologists, radiation oncologists, and urologists are facing a steep learning curve to master the intricacies and nuances of this novel therapeutic strategy. This review article aims to share and discuss the evolving experience in practical aspects of PSMA theranostics.


Microplastics in the Solent estuarine complex, UK: An initial assessment.
Microplastics are known to be an increasing component found within both marine sediments and the water column. This study carried out an initial assessment of the levels of microplastics present within the Solent estuarine complex, focusing specifically on the water column. A plankton net trawl survey was carried out, with samples analysed using visual observation and Fourier Transform Infrared Spectroscopy (FT-IR). The study identified significant quantities of plastics, ranging in shape, with hot spots found at confluence points within the estuary. Though the FT-IR analysis was inconclusive, the nature of the samples indicates the effect of oceanographic conditions on the prevalent types of microplastics found, which in turn identifies key local sources such as wastewater treatment plants and the plastics industry as being the dominant inputs.


Molecular epidemiology of Pseudomonas aeruginosa infections in a cystic fibrosis outpatient clinic.
Chronic respiratory infection by Pseudomonas aeruginosa is a significant determinant in the prognosis of cystic fibrosis patients. Cross-infection between cystic fibrosis patients and the prevalence of P. aeruginosa among them were investigated by microbiological surveillance and RAPD typing of the isolates. A total of 748 samples was cultured, including specimens from the respiratory tract (sputum or throat swabs) and hands of patients and medical staff, resulting in the collection of 86 isolates of P. aeruginosa from 65 samples. Prevalence of P. aeruginosa was 39.3% in respiratory samples, 0.2% on patients' hands and none in the medical staff's hand samples. RAPD typing characterised 51 genotypes and clonal persistence was observed in the majority of patients. These results suggest that cross-infection is not common in the outpatient clinic studied and a common source of acquisition is unlikely.


Unique gene expression and MR T2 relaxometry patterns define chronic murine dextran sodium sulphate colitis as a model for connective tissue changes in human Crohn's disease.
Chronically relapsing inflammation, tissue remodeling and fibrosis are hallmarks of inflammatory bowel diseases. The aim of this study was to investigate changes in connective tissue in a chronic murine model resulting from repeated cycles of dextran sodium sulphate (DSS) ingestion, to mimic the relapsing nature of the human disease. C57BL/6 mice were exposed to DSS in drinking water for 1 week, followed by a recovery phase of 2 weeks. This cycle of exposure was repeated for up to 3 times (9 weeks in total). Colonic inflammation, fibrosis, extracellular matrix proteins and colonic gene expression were studied. In vivo MRI T 2 relaxometry was studied as a potential non-invasive imaging tool to evaluate bowel wall inflammation and fibrosis. Repeated cycles of DSS resulted in a relapsing and remitting disease course, which induced a chronic segmental, transmural colitis after 2 and 3 cycles of DSS with clear induction of fibrosis and remodeling of the muscular layer. Tenascin expression mirrored its expression in Crohn's colitis. Microarray data identified a gene expression profile different in chronic colitis from that in acute colitis. Additional recovery was associated with upregulation of unique genes, in particular keratins, pointing to activation of molecular pathways for healing and repair. In vivo MRI T2 relaxometry of the colon showed a clear shift towards higher T2 values in the acute stage and a gradual regression of T2 values with increasing cycles of DSS. Repeated cycles of DSS exposure induce fibrosis and connective tissue changes with typical features, as occurring in Crohn's disease. Colonic gene expression analysis revealed unique expression profiles in chronic colitis compared to acute colitis and after additional recovery, pointing to potential new targets to intervene with the induction of fibrosis. In vivo T2 relaxometry is a promising non-invasive assessment of inflammation and fibrosis.


Portable Systems for Metered Dispensing of Aggressive Liquids.
Precise metering in liquid dispensing applications often requires application-specific solutions due to incompatibilities of the sensor and actuator components with the dispensed liquids. Some reoccurring challenges are aggressive liquids that would damage the sensors or tubing, the need for sterile liquids while the pumps or sensors cannot be sterilized, or media that can clog the sensor channels. Two different dispensing systems are here presented where the dispensing flow rate or volume is indirectly measured through a coupled pressure change or airflow, thus avoiding contact between the sensor and liquid. The controlled pressure-driven dispensing (cPDD) system builds an overpressure in the liquid reservoir by pumping air and controls the opening of the liquid output valve based on the internal pressure development. The FlowCap system uses a liquid pump on the outlet, controlled by the measured inflow of air to the reservoir. Both systems are designed for compactness and portability and offer independent operation, as well as control and communication, over a wireless interface.


The role of toxins in Clostridium difficile infection.
Clostridium difficile is a bacterial pathogen that is the leading cause of nosocomial antibiotic-associated diarrhea and pseudomembranous colitis worldwide. The incidence, severity, mortality and healthcare costs associated with C. difficile infection (CDI) are rising, making C. difficile a major threat to public health. Traditional treatments for CDI involve use of antibiotics such as metronidazole and vancomycin, but disease recurrence occurs in about 30% of patients, highlighting the need for new therapies. The pathogenesis of C. difficile is primarily mediated by the actions of two large clostridial glucosylating toxins, toxin A (TcdA) and toxin B (TcdB). Some strains produce a third toxin, the binary toxin C. difficile transferase, which can also contribute to C. difficile virulence and disease. These toxins act on the colonic epithelium and immune cells and induce a complex cascade of cellular events that result in fluid secretion, inflammation and tissue damage, which are the hallmark features of the disease. In this review, we summarize our current understanding of the structure and mechanism of action of the C. difficile toxins and their role in disease.


Predicting Risk of Chronic Kidney Disease in Infants and Young Children at Diagnosis of Posterior Urethral Valves: Initial Ultrasound Kidney Characteristics and Validation of Parenchymal Area as Forecasters of Renal Reserve.
There is paucity of validated objective early imaging markers to help predict future renal deterioration in infants with posterior urethral valves. We evaluated the prognostic value of total renal parenchymal area, renal echogenicity and corticomedullary differentiation regarding future development of chronic kidney disease. We analyzed initial postnatal ultrasonographic images from serial posterior urethral valve cases seen at a single tertiary referral center using National Institutes of Health sponsored image processing software. Echogenicity and corticomedullary differentiation were objectively measured as ratios relative to the adjacent liver or spleen and between cortex and medulla. The primary study outcome, renal function at last followup, was dichotomized based on glomerular filtration rate and/or need for renal replacement therapy (dialysis or renal transplantation, stage 5 chronic kidney disease). A total of 75 patients were evaluated, of whom 16 had progression to stage 5 chronic kidney disease after a mean ± SD followup of 64.2 ± 38.9 months. Mean renal parenchymal area was 21.41 cm(2) in patients without and 16 cm(2) in patients with stage 5 chronic kidney disease (p <0.001), and mean corticomedullary differentiation was 1.77 and 1.21, respectively (p <0.001). Bilateral echogenic kidneys were significantly associated with development of stage 5 chronic kidney disease (p = 0.004). The performance of corticomedullary differentiation in predicting stage 5 chronic kidney disease was statistically significant (AUROC 0.881, 95% CI 0.776-0.987, p <0.001). Estimates of renal parenchyma quantity (total renal parenchymal area) and quality (corticomedullary differentiation and renal echogenicity) measured on initial postnatal ultrasound carry prognostic value in determining future risk of stage 5 chronic kidney disease in patients with posterior urethral valves. These data are promising for developing tools to risk stratify patients, counsel parents and customize monitoring protocols.


Psychologic adjustment of the family with a member who has cystic fibrosis.
Seventy-eight percent of the 384 patients 2 years of age and older who were enrolled in the Cystic Fibrosis Clinic at the Hospital for Sick Children participated in this study of family adjustment to cystic fibrosis. Residence with at least one parent at the time of the survey was a condition for inclusion in the study. The effect of the disease on the family was assessed using three questionnaires: an inventory of family problems as perceived by the parents and two standardized measures of family interaction. Measures of behavior and self-concept in each patient and sibling, physical disease parameters for each patient, and demographic characteristics of the families were examined and analyzed in relation to the responses to the questionnaires on family interaction. Families of patients with cystic fibrosis were found to function very well, with normal mean scores on both standardized profiles. Children with cystic fibrosis showed more behavior problems than expected from the test norms, and healthy siblings of children with cystic fibrosis had behavior problem scores midway between those with the disease and norm scores. Measures of self-concept were remarkably similar for patients and siblings and showed levels at or above those for children and young adults in the general population. Parents psychosocial scores were influenced by the patients' physical parameters, but the severity of the disease generally was unrelated to psychosocial scores of patients and siblings. A notable exception was the correlation of emotional disturbance with amount of hospitalization in patients older than 12 years of age.


The role of video-assisted thoracoscopic surgery in the diagnosis of interstitial lung disease.
When a clinical context is indeterminate for idiopathic pulmonary fibrosis (IPF), or a chest high-resolution computed tomography (HRCT) pattern is not indicative of typical or probable usual interstitial pneumonia (UIP) in patients with interstitial lung disease (ILD), surgical lung biopsy should be considered to make a confident diagnosis on the basis of multidisciplinary diagnosis (MDD). The aim of this study was to evaluate the role and safety of video-assisted thoracoscopic surgery (VATS) in patients with ILD. A total of 143 patients with ILD underwent VATS at Toho University Medical Center Omori Hospital between March 2004 and April 2017. We conducted a retrospective study on the usefulness and safety of VATS in the diagnosis of ILD under MDD. The 30-day mortality was 0%. The postoperative complication rate was 12.6%, which included 5 cases of pneumothorax after discharge (3.5%), 4 cases of prolonged air leakage (2.8%), and 2 cases of acute exacerbation (1.4%). Three of 9 cases (33.3%) complicated by pneumothorax after discharge or prolonged air leakage were resected specimens of pleuroparenchymal fibroelastosis (PPFE). Two patients had acute exacerbation, who were ultimately diagnosed as having idiopathic unclassifiable IP and had histologically significant irregular dense fibrosis and numerous fibroblastic foci. The comparison between chest HRCT and histopathological findings revealed 55 cases of possible UIP [UIP (45%), NSIP (25%), and unclassifiable IP (29%)] and 21 cases of inconsistent with UIP [UIP (10%), NSIP (33%), organizing pneumonia (10%), unclassifiable IP (24%), and PPFE (24%)]. VATS can be safely performed to obtain a confident diagnosis for appropriate treatment strategies in patients with ILD.


Expression profiling of stem cell signaling alters with spheroid formation in CD133high/CD44high prostate cancer stem cells.
Cancer stem cells (CSC) isolated from multiple tumor types differentiate in vivo and in vitro when cultured in serum; however, the factors responsible for their differentiation have not yet been identified. The first aim of the present study was to identify CD133high/CD44high DU145 prostate CSCs and compare their profiles with non-CSCs as bulk counterparts of the population. Subsequently, the two populations continued to be three-dimensional multicellular spheroids. Differentiation was then investigated with stem cell-related genomic characteristics. Polymerase chain reaction array analyses of cell cycle regulation, embryonic and mesenchymal cell lineage-related markers, and telomerase reverse transcriptase (TERT) and Notch signaling were performed. Immunohistochemistry of CD117, Notch1, Jagged1, Delta1, Sox2, c-Myc, Oct4, KLF4, CD90 and SSEA1 were determined in CSC and non-CSC monolayer and spheroid subcultures. Significant gene alterations were observed in the CD133high/CD44high population when cultured as a monolayer and continued as spheroid. In this group, marked gene upregulation was determined in collagen type 9 α1, Islet1 and cyclin D2. Jagged1, Delta-like 3 and Notch1 were respectively upregulated genes in the Notch signaling pathway. According to immunoreactivity, the staining density of Jagged1, Sox2, Oct4 and Klf-4 increased significantly in CSC spheroids. Isolated CSCs alter their cellular characterization over the course of time and exhibit a differentiation profile while maintaining their former surface antigens at a level of transcription or translation. The current study suggested that this differentiation process may be a mechanism responsible for the malignant process and tumor growth.


Mechanical action of the intercostal muscles on the ribs.
The external and internal interosseous intercostal muscles were separately stimulated at end-expiratory lung volume in anesthetized dogs. These muscles were all found to elevate the ribs into which they insert. By attaching weights to the ribs, it was determined that the nonlinear compliance of the ribs was responsible for this phenomenon.


A combined experimental and theoretical study of the polar [3 + 2] cycloaddition of electrophilically activated carbonyl ylides with aldehydes and imines.
Numerous 2,5-diaryl-1,3-dioxolane-4,4-dicarbonitriles and 2,4-diphenyl-1,3-oxazolidine-5,5-dicarbonitriles have been synthesized by [3 + 2] cycloaddition reactions between carbonyl ylides generated from epoxides and aldehydes or imines. In contrast to the use of aldehydes (3,4,5-trimethoxybenzaldehyde, piperonal, 1-naphthaldehyde, indole-3-carboxaldehyde, furan-2-carboxaldehyde, and thiophene-2-carboxaldehyde), the reactions performed with imines (N-(phenylmethylene)methanamine, N-(1,3-benzodioxol-5-ylmethylene)propylamine, N-(1,3-benzodioxol-5-ylmethylene)butylamine, and N-(1,3-benzodioxol-5-ylmethylene)benzylamine) proceed diastereoselectively. The effect of microwave irradiation on the outcome of the reaction was studied. The mechanism of these [3 + 2] cycloaddition reactions has been theoretically investigated using DFT methods. These cycloadditions, which have one-step mechanisms, consist of the nucleophilic attack of the aldehyde oxygen or imine nitrogen on the carbonyl ylide. For the reaction with aldehydes, a back-donation effect is responsible for the unexpected reverse charge transfer found at the transition structure. The analysis of the reactivity indexes indicates that the large electrophilic character of the carbonyl ylides induces them to act as strong electrophiles in these polar [3 + 2] cycloaddition reactions.


The regulation of EFG1 in white-opaque switching in Candida albicans involves overlapping promoters.
EFG1, which encodes a trans-acting factor, is expressed as a more abundant 3.2 kb transcript in the white phase and as a less abundant 2.2 kb transcript in the opaque phase of the white-opaque transition in Candida albicans. To understand how alternative phase-specific mRNAs are transcribed from the same gene locus, the 2320 bp upstream region of the gene was functionally characterized by analysing the -activity of deletion derivatives in a luciferase-based reporter system. The white phase-specific promoter contained three discrete sequences involved in white phase-specific activation, between -2022 and -1809 bp (AR1), between -1809 and -1727 bp (AR2) and between -922 and -840 bp (AR3). A higher resolution deletion and mutation analysis of AR2 revealed two regions between -1809 and -1787 bp and between -1764 and -1728 bp that are responsible for AR2 activation. Targeting of promoter constructs to the ectopic ADE2 genomic site and the 3' end of the EFG1 genomic site revealed a positional requirement for white phase-regulated activation specific for the AR2 region of the promoter. Gel mobility shift assays using AR2 revealed a white phase-specific activation complex. No discrete activation sequences were identified in the overlapping promoter of the opaque phase-specific EFG1 transcript. The strength of opaque phase activation was directly proportional to the length of the promoter. Northern analysis excluded the possibility of an opaque phase-specific repressor. These results demonstrate overlapping promoters for white and opaque phase-specific expression of the gene for the transcription factor Efg1, with distinctly different mechanisms of phase-specific activation.


10 years of didactic training for novices in medical education at Charité.
Introduction: Many medical faculties are introducing faculty development programmes to train their teaching staff with the aim of improving student learning performance. Frequently changing parameters within faculties pose a challenge for the sustainable establishment of such programmes. In this paper, we aim to describe facilitating and hindering parameters using the example of the basic teacher training (BTT) course at the Charité - Universtitätsmedizin Berlin (Charité). Project description: After sporadic pilot attempts for university education training, basic teacher training was finally established at the Charité in 2006 for all new teaching staff. An interdisciplinary taskforce at the office for student affairs designed the programme according to the Kern cycle of curriculum development, while the Charité advanced training academy provided the necessary resources. Within ten years more than 900 faculty members have completed the BTT (9% of current active teaching staff at the Charité). The BTT programme underwent several phases (piloting, evaluation, review, personnel and financial boosting), all of which were marked by changes in the staff and organizational framework. Evaluations by participants were very positive, sustainable effects on teaching could be proven to a limited extent. Discussion: Success factors for the establishment of the programme were the institutional framework set by the faculty directors, the commitment of those involved, the support of research grants and the thoroughly positive evaluation by participants. More challenging were frequent changes in parameters and the allocation of incentive resources for other, format-specific training courses (e.g. PBL) as part of the introduction of the new modular curriculum of the Charité. Conclusion: The sustainment of the programme was enabled through strategic institutional steps taken by the faculty heads. Thanks to the commitment and input by those at a working level as well as management level, the basic teacher training course is today an established part of the faculty development programme at the Charité.


Forage feeding to reduce preharvest Escherichia coli populations in cattle, a review.
Although Escherichia coli are commensal organisms that reside within the host gut, some pathogenic strains of E. coli can cause hemorrhagic colitis in humans. The most notable enterohemorrhagic E. coli (EHEC) strain is O157:H7. Cattle are asymptomatic natural reservoirs of E. coli O157:H7, and it has been reported that as many as 30% of all cattle are carriers of this pathogen, and in some circumstances this can be as high as 80%. Feedlot and high-producing dairy cattle are fed large grain rations in order to increase feed efficiency. When cattle are fed large grain rations, some starch escapes ruminal microbial degradation and passes to the hind-gut where it is fermented. EHEC are capable of fermenting sugars released from starch breakdown in the colon, and populations of E. coli have been shown to be higher in grain fed cattle, and this has been correlated with E. coli O157:H7 shedding in barley fed cattle. When cattle were abruptly switched from a high grain (corn) diet to a forage diet, generic E. coli populations declined 1000-fold within 5 d, and the ability of the fecal generic E. coli population to survive an acid shock similar to the human gastric stomach decreased. Other researchers have shown that a switch from grain to hay caused a smaller decrease in E. coli populations, but did not observe the same effect on gastric shock survivability. In a study that used cattle naturally infected with E. coli O157:H7, fewer cattle shed E. coli O157:H7 when switched from a feedlot ration to a forage-based diet compared with cattle continuously fed a feedlot ration. Results indicate that switching cattle from grain to forage could potentially reduce EHEC populations in cattle prior to slaughter; however the economic impact of this needs to be examined.


Voices of the vulnerable: Exploring the livelihood strategies, coping mechanisms and their impact on food insecurity, health and access to health care among Syrian refugees in the Beqaa region of Lebanon.
Lebanon has approximately one million Syrian refugees (SR) registered with the United Nations High Commission on Refugees (UNHCR) and an unknown number of unregistered SR, who cannot benefit from formal assistance. This study aimed to examine the livelihoods, coping strategies, and access to healthcare among SR based on registration status and accompanying formal assistance. A mixed-method approach with more emphasis on the qualitative design was adopted. A purposive convenient sampling approach was used to recruit SR from informal tented settlements (ITS) in the Beqaa region in Lebanon. Data collection included 19 focus group discussions (FGDs) that were conducted with participants, who were further divided into three groups: registered refugees with assistance, registered without assistance and unregistered. Twelve in-depth interviews were conducted with key informants from humanitarian organizations. All interviews and FGDs were audio recorded, transcribed, and thematically analyzed. SR were highly dependent on formal assistance when received, albeit being insufficient. Regardless of registration status, refugees resorted to informal livelihood strategies, including informal employment, child labor, early marriage, and accruing debt. Poor living conditions and food insecurity were reported among all SR. Limited healthcare access and high out-of-pocket costs led to limited use of antenatal care services, prioritizing life-threatening conditions, and resorting to alternative sources of healthcare. Severity of these conditions and their adverse health consequences were especially pronounced among unregistered refugees. Our findings shed light on the economic and health disparities among marginalized SR, with the lack of registration and formal assistance increasing their vulnerability. More tailored and sustainable humanitarian programs are needed to target the most vulnerable and hard-to-reach groups.


Molecular Basis and Clinical Overview of McLeod Syndrome Compared With Other Neuroacanthocytosis Syndromes: A Review.
McLeod syndrome, encoded by the gene XK, is a rare and progressive disease that shares important similarities with Huntington disease but has widely varied neurologic, neuromuscular, and cardiologic manifestations. Patients with McLeod syndrome have a distinct hematologic presentation with specific transfusion requirements. Because of its X-linked location, loss of the XK gene or pathogenic variants in this gene are principally associated with the McLeod blood group phenotype in male patients. The clinical manifestation of McLeod syndrome results from allelic variants of the XK gene or as part of a contiguous gene deletion syndrome involving XK and adjacent genes, including those for chronic granulomatous disease, Duchenne muscular dystrophy, and retinitis pigmentosa. McLeod syndrome typically manifests as neurologic and cardiologic symptoms that evolve in individuals beginning at approximately 40 years of age. Diagnosis of McLeod syndrome encompasses a number of specialties, including neurology and transfusion medicine. However, information regarding the molecular basis of the syndrome is incomplete, and clinical information is difficult to find. The International Society of Blood Transfusion has recently compiled and curated a listing of XK alleles associated with the McLeod phenotype. Of note, McLeod syndrome caused by structural variants as well as those cases diagnosed as part of a contiguous gene deletion syndrome were previously classified under a singular allele designation. This review discusses the clinical manifestations and molecular basis of McLeod syndrome and provides a comprehensive listing of alleles with involvement in the syndrome published to date. This review highlights the clinical diversity of McLeod syndrome and discusses the development of molecular tools to elucidate genetic causes of disease. A more precise and systematic genetic classification is the first step toward correlating and understanding the diverse phenotypic manifestations of McLeod syndrome and may guide clinical treatment of patients and support for affected and carrier family members. This review provides a knowledge base for neurologists, hematologists, and clinical geneticists on this rare and debilitating disease.


Schwann-like cells cultured from human dermal neurofibromas. Immunohistological identification and response to Schwann cell mitogens.
Primary cultures prepared from dermal and plexiform neurofibromas contain Schwann-like cells and fibroblast-like cells. SLC are elongated and bipolar or multipolar. By indirect immunofluorescence light microscopy, living SLC bind antibodies against laminin and against nerve growth factor receptor to their surface, but not antibodies against fibronectin. In these respects, cultured SLC are indistinguishable from cultured human adult Schwann cells. FLC are flat and pleomorphic. By indirect immunofluorescence light microscopy, living FLC bind antibodies against fibronectin but not against laminin or NGFR. In these respects, cultured FLC are indistinguishable from cultured human adult endoneurial fibroblasts. Considerable purification of viable SLC from SLC/FLC mixed cultures can be achieved by flow cytofluorometry using a monoclonal anti-NGFR antibody. Tritiated thymidine radioautography indicated that mitosis of SLC in mixed SLC/FLC cultures prepared from dermal neurofibromas is infrequent in MEM with 10% calf serum, more frequent in RPMI 1640 medium with 15% fetal calf serum. Central nervous system axolemmal fragments (rat or human) elicited a greater than 10-fold SLC proliferative response in mixed SLC/FLC cultures from three of seven dermal neurofibromas (from six patients with neurofibromatosis), but had no effect on SLC mitosis in cultures from the other four dermal neurofibromas. SLC mitosis was inhibited by concentrations of cyclic adenosine 3',5'-monophosphate analogues known to stimulate proliferation of normal rat Schwann cells. Glial growth factor partially purified from bovine pituitaries stimulated SLC mitosis both in SLC/FLC mixed cultures and in cultures of purified SLC. The studies we have described indicate that neurofibroma SLC can be cultured, unequivocally identified in culture by morphological and immunohistological criteria, purified, and stimulated to proliferate by several Schwann cell mitogens. Further quantitative comparisons of the baseline and mitogen-stimulated rates of proliferation of SLC and age-matched control human Schwann cells are needed, however, to determine which of the two alternate pathogenetic mechanisms for formation of neurofibromas mentioned in the introduction is correct.


A case of abdominal aortic pseudoaneurysm associated with right heart mass.
We present a patient with right-sided endocarditis associated with abdominal aortic pseudo-aneurysm presenting only with high fever and pulsating abdominal mass. A higher clinical awareness of aortic pseudoaneurysms associated with intracardiac lesions disease, leading to early computed tomography evaluation and prompt surgical intervention appears to offer the best chance of survival. In this aspect, single-stage surgical treatment of both endocarditis and the aortic pathology is necessary.


Sympathetic predominance is associated with impaired endothelial progenitor cells and tunneling nanotubes in controlled-hypertensive patients.
Early endothelial progenitor cells (early EPC) and late EPC are involved in endothelial repair and can rescue damaged endothelial cells by transferring organelles through tunneling nanotubes (TNT). In rodents, EPC mobilization from the bone marrow depends on sympathetic nervous system activity. Indirect evidence suggests a relation between autonomic derangements and human EPC mobilization. We aimed at testing whether hypertension-related autonomic imbalances are associated with EPC impairment. Thirty controlled-essential hypertensive patients [systolic blood pressure/diastolic blood pressure = 130(120-137)/85(61-88) mmHg; 81.8% male] and 20 healthy normotensive subjects [114(107-119)/75(64-79) mmHg; 80% male] were studied. Mononuclear cells were cultured on fibronectin- and collagen-coated dishes for early EPC and late EPC, respectively. Low (LF)- and high (HF)-frequency components of short-term heart rate variability were analyzed during a 5-min rest, an expiration/inspiration maneuver, and a Stroop color-word test. Modulations of cardiac sympathetic and parasympathetic activities were evaluated by LF/HF (%) and HF power (ms(2)), respectively. In controlled-hypertensive patients, the numbers of early EPC, early EPC that emitted TNT, late EPC, and late EPC that emitted TNT were 41, 77, 50, and 88% lower than in normotensive subjects (P < 0.008), respectively. In controlled-hypertensive patients, late EPC number was positively associated with cardiac parasympathetic reserve during the expiration/inspiration maneuver (rho = 0.45, P = 0.031) and early EPC with brachial flow-mediated dilation (rho = 0.655; P = 0.049); also, late TNT number was inversely related to cardiac sympathetic response during the stress test (rho = -0.426, P = 0.045). EPC exposure to epinephrine or norepinephrine showed negative dose-response relationships on cell adhesion to fibronectin and collagen; both catecholamines stimulated early EPC growth, but epinephrine inhibited late EPC growth. In controlled-hypertensive patients, sympathetic overactivity/parasympathetic underactivity were negatively associated with EPC, suggesting that reducing sympathetic/increasing parasympathetic activation might favor endothelial repair.


Hematuria grading scale: a new tool for gross hematuria.
To develop a new tool for gross hematuria, the Hematuria Grading Scale (HGS), and evaluate its consistency in assessing hematuria samples. The HGS was developed on the basis of an evaluation of sample brightness and saturation using a cyan, magenta, yellow, key (black) color model. Thirty hematuria samples were prepared from human blood by diluting with saline using a standard method. Twenty examiners (5 in each group, including laypeople, nurses, general practitioners, and urologists) participated. Each scored 30 hematuria samples using the HGS under the same conditions without communicating with one another. The intraclass correlation coefficient (ICC) was calculated to assess the reliability of the datasets. Questionnaires for usefulness (Q1) and simplicity (Q2) were obtained from all examiners using a 5-point Likert scale. The ICC for pooled examiner scores showed a high agreement rate (99.7%, 95% confidence interval [CI] 0.996-0.999). ICC values by group were 99.3% (95% CI 0.989-0.997) for laypeople, 98.8% (95% CI 0.980-0.994) for nurses, 99.1% (95% CI 0.984-0.995) for general practitioners, and 99.2% (95% CI 0.987-0.996) for urologists. Mean Q1 and Q2 scores were 4.70 ± 0.66 and 4.30 ± 1.03, respectively, indicating general satisfaction with the HGS among all examiners. Evaluations of gross hematuria using the HGS were in high agreement among examiners of all types, and all examiners found the HGS simple and easy to use. The HGS should be a helpful tool for assessment and communication of gross hematuria.


Genetic variation in brain-derived neurotrophic factor and human fear conditioning.
Brain-derived neurotrophic factor (BDNF) has been implicated in hippocampal-dependent learning processes, and carriers of the Met allele of the Val66Met BDNF genotype are characterized by reduced hippocampal structure and function. Recent nonhuman animal work suggests that BDNF is also crucial for amygdala-dependent associative learning. The present study sought to examine fear conditioning as a function of the BDNF polymorphism. Fifty-seven participants were genotyped for the BDNF polymorphism and took part in a differential-conditioning paradigm. Participants were shocked following a particular conditioned stimulus (CS+) and were also presented with stimuli that ranged in perceptual similarity to the CS+ (20, 40 or 60% smaller or larger than the CS+). The eye blink component of the startle response was measured to quantify fear conditioning; post-task shock likelihood ratings for each stimulus were also obtained. All participants reported that shock likelihood varied with perceptual similarity to the CS+ and showed potentiated startle in response to CS +/- 20% stimuli. However, only the Val/Val group had potentiated startle responses to the CS+. Met allele carrying individuals were characterized by deficient fear conditioning--evidenced by an attenuated startle response to CS+ stimuli. Variation in the BDNF genotype appears related to abnormal fear conditioning, consistent with nonhuman animal work on the importance of BDNF in amygdala-dependent associative learning. The relation between genetic variation in BDNF and amygdala-dependent associative learning deficits is discussed in terms of potential mechanisms of risk for psychopathology.


Three-dimensional image reconstruction of distribution of Pnmt+ cell-derived cells in murine heart.
Elucidating the function of specific cell types in a highly complex multicellular system such as the heart often requires detailed anatomic reconstruction. We recently described a distinctive class of phenylethanolamine n-methyltransferase (Pnmt+) cell-derived cardiomyocytes (PdCMs), a new cardiomyocyte population with a potential endocrine role. In this dataset, a 3D reconstruction was carried out to visualise the distribution of PdCMs throughout the murine heart. Rigid registration (stiff rotation and translation) was applied to properly align the fused heart slice images based on landmarks using TrakEM2, an open source plug-in in Fiji. The registered slices were then analysed and reconstructed using MATLAB (MATLAB®. Version 126.96.36.1992). The final reconstructed 3D volume was 561×866×48 pixels (corresponding to spatial resolutions of 5.8, 8.9 and 2.5 mm in the x-, y- and z-direction respectively), and visualised in Paraview. The reconstruction allows for detailed analyses of morphology, projections and cellular features of different cell types, enabling further geometrical and topological analyses. Image data can be accessed and viewed through Figshare.


Analysis of 118 pesticides in tobacco after extraction with the modified QuEChRS method by LC-MS-MS.
A liquid chromatography-tandem quadrupole mass spectrometry (LC-MS-MS) multi-residue method for the simultaneous target analysis of a wide range of pesticides in tobacco has been developed. Gradient elution has been used in conjunction with positive mode electrospray ionization tandem mass spectrometry to detect up to 118 pesticides in tobacco. The recoveries obtained for each pesticide ranged between 70 and 118% at two spiked concentration levels. Good linear relationships were observed with correlation coefficients r(2) > 0.992 for all analytes. The established method was successfully applied to the determination of pesticide residues in real tobacco samples in order to validate the suitability for routine analysis.


Temperature dependence of blood viscosity in frogs and turtles: effect on heat exchange with environment.
The temperature dependence of the viscosity of blood from frogs and turtles has been assessed for temperatures between 5 and 40 degrees C. Viscosity of turtles' blood was, on average, reduced from 3.50 +/- 0.16 to 2.13 +/- 0.10 cP between 10 and 30 degrees C, a decline of 39%. Even larger changes in viscosity were observed for frogs' blood with viscosity falling from 4.55 +/- 0.32 to 2.55 +/- 0.25 cP over the same temperature range, a change of 44%. Blood viscosity was highly correlated with hematocrit in both species at all temperatures. Viscosity of blood from both frogs and turtles showed a large standard deviation at all temperatures and this was attributed to large individual-to-individual variations in hematocrit. Turtles heat faster than they cool, regardless of whether tests are performed at temperatures above or below the range of thermal preference. The effect of temperature dependence of blood viscosity on heating and cooling rates is demonstrated.


The pro-oxidative riminophenazine B669 neutralizes the inhibitory effects of Mycobacterium tuberculosis on phagocyte antimicrobial activity.
The effects of clofazimine, a riminophenazine antimicrobial agent, and its analogue B669 on phagocyte functions have been investigated. Clofazimine, at concentrations attainable in vivo, and B669, in particular, increased the intracellular killing ability of phagocytes following appropriate cell stimulation. Similarly, nitro blue tetrazolium reduction, hydrogen peroxide production, lysosyme release and hexose monophosphate shunt activity were all increased by treating phagocytes with the riminophenazines. It has previously been shown that a 25 kDa glycolipoprotein derived from Mycobacterium tuberculosis inhibits phagocyte functions associated with phagocyte antimicrobial activity. The present study confirms these observations. A further aspect of the study examined the ability of riminophenazines to reverse the inhibition of phagocyte functions by the 25 kDa mycobacterial fraction. Whilst both riminophenazines were capable of partially but significantly reversing the inhibition due to the mycobacterial fraction, the restorative capacity of B669 was greater than that of clofazimine.


The sensitivity of the stiffness and thickness of a titanium inlay in a cementless PEEK femoral component to the micromotions and bone strain energy density.
Polyetheretherketone (PEEK) has been proposed as alternative material for total knee arthroplasty implants due to its low stiffness, which may reduce stress-shielding. In cementless fixation, a proper primary fixation is required for long-term fixation. Previous research showed that the lower stiffness of a cementless PEEK femoral component results in larger micromotions at the implant-bone interface compared to a cobalt-chrome femoral component. A titanium inlay on the PEEK implant surface may improve the primary fixation while maintaining the favourable stiffness properties. Therefore, the effect of thickness and stiffness of a titanium inlay on the primary fixation and stress-shielding was investigated. A finite element model of the femur and femoral component was created with five titanium inlay variants. The micromotions and strain energy density (SED) were quantified as outcome measures. The distal thin - proximal thick variant showed the largest resulting micromotions (51.2 µm). Relative to the all-PEEK femoral component, the addition of a titanium inlay reduced the micromotions with 30 % to 40 % without considerably affecting the stress-shielding capacity (strain energy difference of 6 % to 10 %). Differences in micromotions (43.0-51.2 µm) and SED between the variants were relatively small. In conclusion, the addition of a titanium inlay could lead to a reduction of the micromotions without substantially affecting the SED distribution.


Discovery of fragments inducing conformational effects in dynamic proteins using a second-harmonic generation biosensor.
Biophysical screening of compound libraries for the identification of ligands that interact with a protein is efficient, but does typically not reveal if (or how) ligands may interfere with its functional properties. For this a biochemical/functional assay is required. But for proteins whose function is dependent on a conformational change, such assays are typically complex or have low throughput. Here we have explored a high-throughput second-harmonic generation (SHG) biosensor to detect fragments that induce conformational changes upon binding to a protein in real time and identify dynamic regions. Multiwell plate format SHG assays were developed for wild-type and six engineered single-cysteine mutants of acetyl choline binding protein (AChBP), a homologue to ligand gated ion channels (LGICs). They were conjugated with second harmonic-active labels via amine or maleimide coupling. To validate the assay, it was confirmed that the conformational changes induced in AChBP by nicotinic acetyl choline receptor (nAChR) agonists and antagonists were qualitatively different. A 1056 fragment library was subsequently screened against all variants and conformational modulators of AChBP were successfully identified, with hit rates from 9-22%, depending on the AChBP variant. A subset of four hits was selected for orthogonal validation and structural analysis. A time-resolved grating-coupled interferometry-based biosensor assay confirmed the interaction to be a reversible 1-step 1 : 1 interaction, and provided estimates of affinities and interaction kinetic rate constants (K D = 0.28-63 μM, k a = 0.1-6 μM-1 s-1, k d = 1 s-1). X-ray crystallography of two of the fragments confirmed their binding at a previously described conformationally dynamic site, corresponding to the regulatory site of LGICs. These results reveal that SHG has the sensitivity to identify fragments that induce conformational changes in a protein. A selection of fragment hits with a response profile different to known LGIC regulators was characterized and confirmed to bind to dynamic regions of the protein.


Mechanism-Guided Design of Photocatalysts for CO2 Reduction toward Multicarbon Products.
Photocatalytic reduction of CO2 to high value-added multicarbon (C2+) products is an important way to achieve sustainable production of green energy but limited by the low efficiency of catalysts. One fundamental issue lies in the high complexity of catalyst structure and reaction process, making the rational catalyst design and targeted performance optimization a grand challenge. Herein, we performed a mechanism-guided design of photocatalysts for CO2 reduction by using the experimentally reported Cu doped TiO2 (Cu-TiO2) with high C3H8 selectivity and well-defined structure as the prototype. Our mechanistic study highlights three key factors for C3H8 formation, i.e., formation of double O vacancies (Vdi-O) for selectivity, C-C coupling for activity, and Vdi-O recovery for durability. More importantly, Vdi-O formation/recovery and C-C coupling are negatively correlated, indicating that ideal candidates should achieve a balance between oxygen vacancy (VO) formation and C-C coupling. On this basis, TiO2 with the doping of two adjacent Cu atoms (Cu-Cu-TiO2) was designed with enhanced performance for CO2 photoreduction toward C3H8. Furthermore, a simple descriptor (Nμ, "effective d electron number") based on inherent atomic properties was constructed to uncover the underlying causes of the performance variation of different systems. These results provide new insights into the "structure-performance" relation of metal oxide-based photocatalysts, thus offering useful strategies for the rational design of excellent catalysts for CO2 photoreduction.


Small Performance Effects of a Practical Mixed-Methods Cooling Strategy in Elite Team Sport Athletes.
Purpose: The ingestion of ice slurry and application of ice towels can elicit favorable physiological, perceptual, and performance benefits when used individually; however, the combined use and effectiveness of these practical cooling strategies have not been assessed using a sport-specific performance test, based on actual match demands, in an elite team sport context. Methods: Ten non-heat acclimated elite male rugby sevens athletes undertook two cycling heat response tests (HRT) designed to be specific to the demands of rugby sevens in hot conditions (35°C, 80% rH). In a crossover design, the HRTs were conducted with (COOLING) and without (HOT) the combined use of internal (ice slushy ingestion) and external (application of ice towels to the head, neck, and face) pre- and per-cooling strategies. Physiological, perceptual, and performance variables were monitored throughout each HRT. Results: COOLING resulted in reductions in mean tympanic temperature (-0.4 ± 0.2°C; d = 1.18); mean heart rate (-5 ± 8 bpm; d = 0.53); thermal discomfort (-0.5 ± 0.9 AU; d = 0.48); and thirst sensation (-1.0 ± 1.1 AU; d = 0.61) during the HRT. COOLING also resulted in a small increase in 4-min time trial power output (by 7 ± 33 W, ~3%; d = 0.35) compared to HOT. Discussion: A combination of internal and external pre- and per-cooling strategies can result in a range of small physiological, perceptual, and performance benefits during a rugby sevens specific HRT, compared to undertaking no cooling. Practitioners should include such strategies when performing in hot conditions.


Direct transactivator-transcription factor IID (TFIID) contacts drive yeast ribosomal protein gene transcription.
Transcription factor IID (TFIID) plays a key role in regulating eukaryotic gene expression by directly binding promoters and enhancer-bound transactivator proteins. However, the precise mechanisms and outcomes of transactivator-TFIID interaction remain unclear. Transcription of yeast ribosomal protein genes requires TFIID and the DNA-binding transactivator Rap1. We have previously shown that Rap1 directly binds to the TFIID complex through interaction with its TATA-binding protein-associated factor (Taf) subunits Taf4, -5, and -12. Here, we identify and characterize the Rap1 binding domains (RBDs) of Taf4 and Taf5. These RBDs are essential for viability but dispensable for Taf-Taf interactions and TFIID stability. Cells expressing altered Rap1 binding domains exhibit conditional growth, synthetic phenotypes when expressed in combination or with altered Rap1, and are selectively defective in ribosomal protein gene transcription. Taf4 and Taf5 proteins with altered RBDs bind Rap1 with reduced affinity. We propose that collectively the Taf4, Taf5, and Taf12 subunits of TFIID represent the physical and functional targets for Rap1 interaction and, furthermore, that these interactions drive ribosomal protein gene transcription.