The Number of Pulses Needed to Measure Corticospinal Excitability by Navigated Transcranial Magnetic Stimulation: Eyes Open vs. Close Condition.
Objective: Motor evoked potentials (MEPs) obtained by transcranial magnetic stimulation (TMS) enable measures of the corticospinal excitability (CSE). However the reliability of TMS-derived CSE measures is suboptimal due to appreciable pulse-to-pulse MEP amplitude variability. We thus calculated how many TMS-derived MEPs will be needed to obtain a reliable CSE measure in awake adult subjects, in the eyes open (EO) and eyes closed (EC) conditions. Methods: Twenty healthy adults (70% male) received 40 consecutive navigated TMS pulses (120% resting motor threshold, RMT) in the EO or EC conditions on two separate days in randomized order. Results: For either the EO or EC condition, the probability that the 95% confidence interval (CI) derived from consecutive MEP amplitude measured included the true CSE, increased when the number of consecutive stimuli increased (EO: p = 0.05; EC: p = 0.001). No significant effect of RMT, Mini-Mental State Examination (MMSE) score, or gender on the CSE estimates was identified. At least 34 consecutive stimuli were required to obtain a most reliable CSE estimate in the EO condition and 31 in the EC condition. Conclusion: Our findings indicate that >30 consecutive MEPs may be necessary in order to obtain a CSE measure in healthy adults.


Neuron-specific cholinergic modulation of a forebrain song control nucleus.
Cholinergic activation profoundly affects vertebrate forebrain networks, but pathway, cell type, and modality specificity remain poorly understood. Here we investigated cell-specific cholinergic modulation of neurons in the zebra finch forebrain song control nucleus HVC using in vitro whole cell recordings. The HVC contains projection neurons that exclusively project to either another song motor nucleus RA (robust nucleus of the arcopallium) (HVC-RAn) or the basal ganglia Area X (HVC-Xn) and these populations are synaptically coupled by a network of GABAergic interneurons. Among HVC-RAn, we observed two physiologically distinct classes that fire either phasically or tonically to injected current. Muscarine excited phasic HVC-RAn and most HVC-Xn. Effects were observed under conditions of blockade of fast synaptic transmission and were reversed by atropine. In contrast, unlike what is commonly observed in mammalian systems, HVC interneurons were inhibited by muscarine and these effects were reversed by atropine. Thus cholinergic modulation reconfigures the HVC network in a more complex fashion than that implied by monolithic "gating." The two projection pathways are decoupled through suppression of the inhibitory network that links them, whereas each is simultaneously predominantly excited. We speculate that fluctuating cholinergic tone in HVC could modulate the interaction of song motor commands with basal ganglia circuitry associated with song perception and modification. Furthermore, if the in vitro distinction between RA-projecting neurons that we observed is also present in vivo, then the song system motor pathway exhibits greater physiological diversity than has been commonly assumed.


Transport of cytokinins mediated by purine transporters of the PUP family expressed in phloem, hydathodes, and pollen of Arabidopsis.
Nucleobases and derivatives like cytokinins and caffeine are translocated in the plant vascular system. Transport studies in cultured Arabidopsis cells indicate that adenine and cytokinin are transported by a common H+-coupled high-affinity purine transport system. Transport properties are similar to that of Arabidopsis purine transporters AtPUP1 and 2. When expressed in yeast, AtPUP1 and 2 mediate energy-dependent high-affinity adenine uptake, whereas AtPUP3 activity was not detectable. Similar to the results from cell cultures, purine permeases (PUP) mediated uptake of adenine can be inhibited by cytokinins, indicating that cytokinins are transport substrates. Direct measurements demonstrate that AtPUP1 is capable of mediating uptake of radiolabeled trans-zeatin. Cytokinin uptake is strongly inhibited by adenine and isopentenyladenine but is poorly inhibited by 6-chloropurine. A number of physiological cytokinins including trans- and cis-zeatin are also efficient competitors for AtPUP2-mediated adenine uptake, suggesting that AtPUP2 is also able to mediate cytokinin transport. Furthermore, AtPUP1 mediates transport of caffeine and ribosylated purine derivatives in yeast. Promoter-reporter gene studies point towards AtPUP1 expression in the epithem of hydathodes and the stigma surface of siliques, suggesting a role in retrieval of cytokinins from xylem sap to prevent loss during guttation. The AtPUP2 promoter drives GUS reporter gene activity in the phloem of Arabidopsis leaves, indicating a role in long-distance transport of adenine and cytokinins. Promoter activity of AtPUP3 was only found in pollen. In summary, three closely related PUPs are differentially expressed in Arabidopsis and at least two PUPs have properties similar to the adenine and cytokinin transport system identified in Arabidopsis cell cultures.


Toxicity, elimination, and metabolism of 10-ethyl-10-deazaaminopterin in rats and dogs.
10-Ethyl-10-deazaaminopterin (10-EdAM) is an antifolate compound with greater therapeutic activity than methotrexate against transplanted tumors in mice. When given weekly for 3 weeks, the 10% lethal dose in rats was 125 mg/kg (i.p.) and in dogs it was 2.5 mg/kg (i.v.). The major histopathological findings in intoxicated animals were damage to the mucosa of the gastrointestinal tract in rats and dogs and hypocellularity of the marrow in rats. The elimination of 50 mg/kg of 10-EdAM from the plasma of rats was triexponential with a terminal phase t1/2 of 18.5 h but a mean residence time of 0.7 h. The primary route of elimination in rats was biliary secretion of parent compound and eventual excretion of the parent compound and the deglutamate metabolite in the feces; the 7-hydroxy metabolite was also present in plasma, bile, and feces. Biliary elimination was independent of dose over a 5-fold range. The elimination of 10-EdAM from the plasma of dogs was also triexponential with a mean terminal phase t1/2 of 9.1 h and a mean residence time of 2.5 h; nonrenal clearance was the primary route of elimination. The pharmacokinetic parameters were independent of dose over the range of 0.25 to 5.0 mg/kg. High tissue concentrations of 10-EdAM were observed initially in liver, kidney, and small intestine of rats, while concentrations in bone marrow were low. Some polyglutamate formation was observed in these tissues as early as 0.5 h after drug administration but declined over 72 h.


A technique for the quantitative evaluation of dose distributions.
The commissioning of a three-dimensional treatment planning system requires comparisons of measured and calculated dose distributions. Techniques have been developed to facilitate quantitative comparisons, including superimposed isodoses, dose-difference, and distance-to-agreement (DTA) distributions. The criterion for acceptable calculation performance is generally defined as a tolerance of the dose and DTA in regions of low and high dose gradients, respectively. The dose difference and DTA distributions complement each other in their useful regions. A composite distribution has recently been developed that presents the dose difference in regions that fail both dose-difference and DTA comparison criteria. Although the composite distribution identifies locations where the calculation fails the preselected criteria, no numerical quality measure is provided for display or analysis. A technique is developed to unify dose distribution comparisons using the acceptance criteria. The measure of acceptability is the multidimensional distance between the measurement and calculation points in both the dose and the physical distance, scaled as a fraction of the acceptance criteria. In a space composed of dose and spatial coordinates, the acceptance criteria form an ellipsoid surface, the major axis scales of which are determined by individual acceptance criteria and the center of which is located at the measurement point in question. When the calculated dose distribution surface passes through the ellipsoid, the calculation passes the acceptance test for the measurement point. The minimum radial distance between the measurement point and the calculation points (expressed as a surface in the dose-distance space) is termed the gamma index. Regions where gamma > 1 correspond to locations where the calculation does not meet the acceptance criteria. The determination of gamma throughout the measured dose distribution provides a presentation that quantitatively indicates the calculation accuracy. Examples of a 6 MV beam penumbra are used to illustrate the gamma index.


Interrater Reliability of Experts in Identifying Interictal Epileptiform Discharges in Electroencephalograms.
The validity of using electroencephalograms (EEGs) to diagnose epilepsy requires reliable detection of interictal epileptiform discharges (IEDs). Prior interrater reliability (IRR) studies are limited by small samples and selection bias. To assess the reliability of experts in detecting IEDs in routine EEGs. This prospective analysis conducted in 2 phases included as participants physicians with at least 1 year of subspecialty training in clinical neurophysiology. In phase 1, 9 experts independently identified candidate IEDs in 991 EEGs (1 expert per EEG) reported in the medical record to contain at least 1 IED, yielding 87 636 candidate IEDs. In phase 2, the candidate IEDs were clustered into groups with distinct morphological features, yielding 12 602 clusters, and a representative candidate IED was selected from each cluster. We added 660 waveforms (11 random samples each from 60 randomly selected EEGs reported as being free of IEDs) as negative controls. Eight experts independently scored all 13 262 candidates as IEDs or non-IEDs. The 1051 EEGs in the study were recorded at the Massachusetts General Hospital between 2012 and 2016. Primary outcome measures were percentage of agreement (PA) and beyond-chance agreement (Gwet κ) for individual IEDs (IED-wise IRR) and for whether an EEG contained any IEDs (EEG-wise IRR). Secondary outcomes were the correlations between numbers of IEDs marked by experts across cases, calibration of expert scoring to group consensus, and receiver operating characteristic analysis of how well multivariate logistic regression models may account for differences in the IED scoring behavior between experts. Among the 1051 EEGs assessed in the study, 540 (51.4%) were those of females and 511 (48.6%) were those of males. In phase 1, 9 experts each marked potential IEDs in a median of 65 (interquartile range [IQR], 28-332) EEGs. The total number of IED candidates marked was 87 636. Expert IRR for the 13 262 individually annotated IED candidates was fair, with the mean PA being 72.4% (95% CI, 67.0%-77.8%) and mean κ being 48.7% (95% CI, 37.3%-60.1%). The EEG-wise IRR was substantial, with the mean PA being 80.9% (95% CI, 76.2%-85.7%) and mean κ being 69.4% (95% CI, 60.3%-78.5%). A statistical model based on waveform morphological features, when provided with individualized thresholds, explained the median binary scores of all experts with a high degree of accuracy of 80% (range, 73%-88%). This study's findings suggest that experts can identify whether EEGs contain IEDs with substantial reliability. Lower reliability regarding individual IEDs may be largely explained by various experts applying different thresholds to a common underlying statistical model.


Concise Review: early embryonic erythropoiesis: not so primitive after all.
In the developing embryo, hematopoiesis begins with the formation of primitive erythroid cells (EryP), a distinct and transient red blood cell lineage. EryP play a vital role in oxygen delivery and in generating shear forces necessary for normal vascular development. Progenitors for EryP arise as a cohort within the blood islands of the mammalian yolk sac at the end of gastrulation. As a strong heartbeat is established, nucleated erythroblasts begin to circulate and to mature in a stepwise, nearly synchronous manner. Until relatively recently, these cells were thought to be "primitive" in that they seemed to more closely resemble the nucleated erythroid cells of lower vertebrates than the enucleated erythrocytes of mammals. It is now known that mammalian EryP do enucleate, but not until several days after entering the bloodstream. I will summarize the common and distinguishing characteristics of primitive versus definitive (adult-type) erythroid cells, review the development of EryP from the emergence of their progenitors through maturation and enucleation, and discuss pluripotent stem cells as models for erythropoiesis. Erythroid differentiation of both mouse and human pluripotent stem cells in vitro has thus far reproduced early but not late red blood cell ontogeny. Therefore, a deeper understanding of cellular and molecular mechanisms underlying the differences and similarities between the embryonic and adult erythroid lineages will be critical to improving methods for production of red blood cells for use in the clinic.


Warm fresh whole blood transfusion for severe hemorrhage: U.S. military and potential civilian applications.
The objective of this study was to review the history and current literature regarding the benefits and risks of warm fresh whole blood transfusion to include recent U.S. Army research from Afghanistan and Iraq. We also discuss current indications for its use as well as potential civilian applications for large-scale disasters. The use of warm fresh whole blood currently only persists in emergency life-threatening scenarios when tested stored blood components are not available. Recent combat operations in Afghanistan and Iraq have redirected attention on the benefits and risks of warm fresh whole blood for life-threatening injuries in casualties. Between March 2003 and July 2007, over 6000 units of warm fresh whole blood have been transfused in Afghanistan and Iraq by U.S. medical providers to patients with life-threatening traumatic injuries with hemorrhage. Preliminary results in approximately 500 patients with massive transfusion indicate that the amount of fresh warm whole blood transfused is independently associated with improved 48-hr and 30-day survival and the amount of stored red blood cells is independently associated with decreased 48-hr and 30-day survival for patients with traumatic injuries that require massive transfusion. Risks of warm fresh whole blood transfusion include the transmission of infectious agents and the potential for microchimerism. For patients with life-threatening hemorrhage at risk for massive transfusion, if complete component therapy is not available or not adequately correcting coagulopathy, the risk:benefit ratio of warm fresh whole blood transfusion favors its use. In addition, recent evidence suggests that there is potential for warm fresh whole blood to be more efficacious than stored component therapy that includes stored red blood cells in critically ill patients requiring massive transfusion. Efforts must continue to improve the safety of warm fresh whole blood transfusion for patients when it is required in emergency situations. U.S. civilian disaster agencies are preparing guidelines for its use in massive casualty scenarios and prospective, randomized trials are about to start to determine whether stored warm fresh (<24 hrs) whole blood improves outcomes compared with standard stored component therapy.


Intrinsic signals regulate the initial steps of myogenesis in vertebrates.
In vertebrates, despite the evidence that extrinsic factors induce myogenesis in naive mesoderm, other experiments argue that the initiation of the myogenic program may take place independent of these factors. To resolve this discrepancy, we have re-addressed this issue, using short-term in vivo microsurgery and culture experiments in chick. Our results show that the initial expression of the muscle-specific markers Myf5 and MyoD is regulated in a mesoderm-autonomous fashion. The reception of a Wnt signal is required for MyoD, but not Myf5 expression; however, we show that the source of the Wnt signal is intrinsic to the mesoderm. Gain- and loss-of-function experiments indicate that Wnt5b, which is expressed in the presomitic mesoderm, represents the MyoD-activating cue. Despite Wnt5b expression in the presomitic mesoderm, MyoD is not expressed in this tissue: our experiments demonstrate that this is due to a Bmp inhibitory signal that prevents the premature expression of MyoD before somites form. Our results indicate that myogenesis is a multistep process which is initiated prior to somite formation in a mesoderm-autonomous fashion; as somites form, influences from adjacent tissues are likely to be required for maintenance and patterning of early muscles.


Induction of coiling in tendrils by auxin and carbon dioxide.
Symmetric application of indole-3-acetic acid, CO(2), or, to a lesser extent, ethylene can substitute for the contact stimulus in inducing coiling in the tendrils of Marah fabaceus. In the case of auxin, treatment of the apical few millimeters results in strong, permanent coiling throughout the length of the tendril. The speed of the response to CO(2) is comparable to that to tactile stimuli. A possible mechanism for thigmotropism is outlined.


Resting metabolic rate and anthropometry in older people: a comparison of measured and calculated values.
Accurate assessment of energy expenditure and anthropometry in older people is important for targeted nutritional support. The present study aimed to compare measured and calculated resting metabolic rate (m-RMR and c-RMR) and measured, calculated and estimated weight and height in older people aged ≥70 years. Participants were healthy older people aged ≥70 years. Indirect calorimetry using a ventilated hood calorimeter was performed for 30 min on fasted participants, and was compared with c-RMR, as calculated using six commonly used equations. Measured, calculated and estimated height and weight were compared. Subjects comprised 14 males and 20 females and mean (SD) m-RMR was 5243 (845) kJ day(-1) [1253 (202) kcal day(-1) ]. The Mifflin St-Jeor equation was the most consistently accurate, with the smallest mean difference between m-RMR and c-RMR of 58 (553) kJ day(-1) [14 (132) kcal day(-1) ] and c-RMR was within 10% of m-RMR in the greatest number of participants (n = 24; 70%). The Schofield equation was among the least accurate in this age group. In older males, self-reported height and weight were accurate, whereas, in females or those unable to self-report height, ulna length was the most accurate alternative to measured height. Current equations used to calculate RMR in older people have inaccuracies, although the Mifflin St-Jeor equation was most accurate. Future studies should investigate the validity, reliability, cost and practicality of using fat free mass as an item in novel equations to calculate RMR in this age group. Self-reported height and weight in males, and height calculated from ulna length in females, were the most accurate alternatives to measured values in the present study.


Psychiatric Diagnoses Among HIV-Exposed and HIV-Unexposed Uninfected Children: A Danish Nationwide Cohort Study.
This nationwide registry-based cohort study aimed to compare the risk of psychiatric diagnoses among HIV-exposed uninfected (HEU) children with a matched comparison group of HIV-unexposed uninfected (HUU) children, born in Denmark. We hypothesized that HEU children had an increased risk of psychiatric diagnoses and that this increased risk may differ by sex and age. All HEU children born in Denmark between year 2000 and 2020 were included. Each HEU child was matched by year of birth, maternal age at birth, and maternal immigration status to 10 HUU children. The primary outcome was risk of any psychiatric diagnosis (International Classification of Diseases, 10th Revision F00-F99). Incidence rate ratios (IRRs) were estimated using Poisson regression. Analyses stratifying by sex and age were also conducted. In total, 550 HEU children and 5500 HUU children were included. HEU children had an increased risk of any psychiatric disorder [IRR 1.45; 95% confidence interval (CI): 1.04-2.04] in the unadjusted analysis, but in the adjusted analysis, the risk was only significant for children aged 6-11 years [adjusted incidence rate ratio (aIRR) 1.93; 95% CI: 1.14-3.28]. Stratifying by sex, girls aged 6-11 years had an increased risk of any psychiatric disorder (aIRR 3.04; 95% CI: 1.27-7.28), while boys had an increased risk at age 12-20 years (aIRR 2.47; 95% CI: 1.18-5.17). In conclusion, HEU girls aged 6-11 years and HEU boys aged 12-20 years had an increased risk of any psychiatric disorder compared with HUU girls and boys, respectively. These findings highlight the importance of addressing the mental health needs of HEU children/adolescents.


Osmolality predictive models of different polymers as tools in parenteral and ophthalmic formulation development.
During the development of parenteral dosage forms, different physicochemical studies are required to ensure stable, effective and safe formulations. The osmolality of this kind of dosage forms should bear a close similarity to the body fluids to prevent local irritation, pain or even more significant side effects like endothelial damage. The osmotic studies performed in Polyethylene glycol 400 (PEG 400), Polyethylene glycol 4000 (PEG 4000), Poloxamer 407 (P407), Sodium Hyaluronate (SH), Chondroitin Sulphate Sodium (CS), Cremophor RH 40 (CRE40) and Polyvinyl alcohol (PVA) aqueous solutions, showed that the theoretical determination of the osmolality based on their molecular weight as the only determinant factor did not agree with the values obtained by the measurement of colligative properties such as the freezing point depression. The data obtained from this study and its analysis, provided predictive equations that can be used as tools in the primary development to estimate formulation's osmolality at different concentrations; and its evolution over a period at the hypothetical worst-case scenario of storage temperature.


Agreement between Mantoux skin testing and QuantiFERON-TB assay using dual mycobacterial antigens in current and former injection drug users.
Individuals infected with non-tuberculous mycobacteria may elicit false-positive reactions on tuberculin skin testing. The QuantiFERON-TB (QFT) assay utilizes tuberculin and M. avium antigens and, therefore, may be more specific for latent tuberculosis infection. The objective of this study was to investigate the agreement between the QFT and single and dual antigen skin testing for detecting latent M. tuberculosis and assess the impact of cross-reactions from latent infection with other mycobacteria in inner-city injection drug users, a population at high risk for tuberculosis. We studied the agreement of results from skin testing using tuberculin and purified protein derivative-Battey (PPD-B) with the QFT test using tuberculin and Mycobacterium avium sensitin (MAS) in 48 HIV-seronegative injection drug users. The agreement between skin testing and the QFT assay for tuberculin was 73% (kappa = 0.45) and for PPD-B/MAS was 63% (kappa = 0.12). Agreement between skin test tuberculin dominance (tuberculin reaction > or =5 mm greater than PPD-B) and QFT tuberculin dominance (proportional difference between MAS and tuberculin reaction of > or =10%) was 75% (kappa = 0.53). All subjects tuberculin dominant by skin test were also QFT positive for tuberculin. Agreement between skin test Battey dominance and QFT avium dominance was 83% (kappa = 0.12). Results from the QFT assay and skin testing demonstrated moderate concordance in identifying subjects with latent tuberculous infection, and use of dual antigens did not appreciably improve the agreement between the two methods.


[Nontraumatic hemorrhagic stroke in children after the neonatal period].
The aim of this study was to determine the incidence, clinical features and outcome of nontraumatic hemorrhagic stroke in children. We performed a retrospective study of children with nontraumatic hemorrhagic stroke (codes 430-432 of the International Classification of Disease, 9th revision). Between January 1982 and December 1995, twenty-one children less than 16 years of age with nontraumatic hemorrhagic stroke were admitted to our hospital. The incidence rate for hemorrhagic stroke was 2.3 cases per 100,000 per year which constituted 67.7% of all strokes in children. There was a slight predominance of males (1.62:1). Onset of symptoms was sudden in 61.9% of cases and gradual in 38%. The most commo symptoms were altered consciousness (71.4%) and hemiparesis (61.9%). Vascular malformations constituted the largest etiological group (42.8%) followed by the hemorrhages into cerebral tumors (9.5%). In 28.5% of the cases no cause could be found. Surgery was carried out in 38% of the cases. There was an overall mortality of 19% and a post-operative mortality of 25%. Hemorrhagic stroke is a rare event in childhood. Prompt surgical management decreases the mortality rate and improves the outcome of these patients.


Interpretable Representation Learning for Healthcare via Capturing Disease Progression through Time.
Various deep learning models have recently been applied to predictive modeling of Electronic Health Records (EHR). In medical claims data, which is a particular type of EHR data, each patient is represented as a sequence of temporally ordered irregularly sampled visits to health providers, where each visit is recorded as an unordered set of medical codes specifying patient's diagnosis and treatment provided during the visit. Based on the observation that different patient conditions have different temporal progression patterns, in this paper we propose a novel interpretable deep learning model, called Timeline. The main novelty of Timeline is that it has a mechanism that learns time decay factors for every medical code. This allows the Timeline to learn that chronic conditions have a longer lasting impact on future visits than acute conditions. Timeline also has an attention mechanism that improves vector embeddings of visits. By analyzing the attention weights and disease progression functions of Timeline, it is possible to interpret the predictions and understand how risks of future visits change over time. We evaluated Timeline on two large-scale real world data sets. The specific task was to predict what is the primary diagnosis category for the next hospital visit given previous visits. Our results show that Timeline has higher accuracy than the state of the art deep learning models based on RNN. In addition, we demonstrate that time decay factors and attentions learned by Timeline are in accord with the medical knowledge and that Timeline can provide a useful insight into its predictions.


APASL HCV guidelines of virus-eradicated patients by DAA on how to monitor HCC occurrence and HBV reactivation.
In the direct-acting antiviral (DAA) era for hepatitis C virus (HCV) infection, sustained virological response (SVR) is very high, but close attention must be paid to the possible occurrence of hepatocellular carcinoma (HCC) and reactivation of hepatitis B virus (HBV) in patients with co-infection who achieved SVR in short term. HCC occurrence was more often observed in patients with previous HCC history. We found occurrence of HCC in 178 (29.6%) of 602 patients with previous HCC history (15.4 months mean follow-up post-DAA initiation) but, in contrast, in only 604 (1.3%) of 45,870 patients without previous HCC history (18.2 months mean follow-up). Thus, in these guidelines, we recommend the following: in patients with previous HCC history, surveillance at 4-month intervals for HCC by ultrasonography (US) and tumor markers should be performed. In patients without previous HCC history, surveillance at 6- to 12-month intervals for HCC including US is recommended until the long-term DAA treatment effects, especially for the resolution of liver fibrosis, are confirmed. This guideline also includes recommendations on how to follow-up patients who have been infected with both HCV and HBV. When HCV was eradicated in these HBsAg-positive patients or patients with previous HBV infection (anti-HBc and/or anti-HBs-positive), it was shown that HBV reactivation or HBV DNA reappearance was observed in 67 (41.4%) of 162 or 12 (0.9%) of 1317, respectively. For these co-infected patients, careful attention should be paid to HBV reactivation for 24 weeks post-treatment.


Hierarchical Features Driven Residual Learning for Depth Map Super-Resolution.
Rapid development of affordable and portable consumer depth cameras facilitates the use of depth information in many computer vision tasks such as intelligent vehicles and 3D reconstruction. However, depth map captured by low-cost depth sensors (e.g., Kinect) usually suffers from low spatial resolution, which limits its potential applications. In this paper, we propose a novel deep network for depth map super-resolution (SR), called DepthSR-Net. The proposed DepthSR-Net automatically infers a high resolution (HR) depth map from its low resolution (LR) version by hierarchical features driven residual learning. Specifically, DepthSR-Net is built on a residual U-Net deep network architecture. Given LR depth map, we first obtain the desired HR by bicubic interpolation upsampling, and then construct an input pyramid to achieve multiple level receptive fields. Next, we extract hierarchical features from the input pyramid, intensity image, and encoder-decoder structure of UNet. Finally, we learn the residual between the interpolated depth map and the corresponding HR one using the rich hierarchical features. The final HR depth map is achieved by adding the learned residual to the interpolated depth map. We conduct an ablation study to demonstrate the effectiveness of each component in the proposed network. Extensive experiments demonstrate that the proposed method outperforms the state-of-the-art methods. Additionally, the potential usage of the proposed network in other low-level vision problems is discussed.


Regulation of Exopolysaccharide Production by ProE, a Cyclic-Di-GMP Phosphodiesterase in Pseudomonas aeruginosa PAO1.
The ubiquitous second messenger c-di-GMP is involved in regulation of multiple biological functions including the important extracellular matrix exopolysaccharides (EPS). But how c-di-GMP metabolic proteins influence EPS and their enzymatic properties are not fully understood. Here we showed that deletion of proE, which encodes a protein with GGDEF-EAL hybrid domains, significantly increased the transcriptional expression of the genes encoding EPS production in Pseudomonas aeruginosa PAO1 and changed the bacterial colony morphology. Our data showed that ProE is a very active phosphodiesterase (PDE), with a high enzyme activity in degradation of c-di-GMP. Interestingly, the optimal activity of ProE was found in the presence of Co2+, unlike other PDEs that commonly rely on Mg2+ or Mn2+ for best performance. Furthermore, we identified three widely conserved novel residues that are critical for the function of ProE through site-directed mutagenesis. Subsequent study showed that ProE, together with other three key PDEs, i.e., RbdA, BifA, and DipA regulate the EPS production in P. aeruginosa PAO1. Moreover, by using the GFP-fusion approach, we observed that these four EPS associated-PDEs showed a polar localization pattern in general. Taken together, our data unveil the molecular mechanisms of ProE in regulation of EPS production, and provide a new insight on its enzymatic properties in degradation of c-di-GMP.


Demonstrating the Protective Efficacy of the Novel Fluoroquinolone Finafloxacin against an Inhalational Exposure to Burkholderia pseudomallei.
Burkholderia pseudomallei is the causative agent of melioidosis, a serious disease endemic in Southeast Asia and Northern Australia. Antibiotic treatment is lengthy and relapse often occurs. Finafloxacin is a novel fluoroquinolone with increased antibacterial activity in acidic conditions in contrast to other fluoroquinolones which demonstrate reduced activity at a lower pH. Therefore, finafloxacin may have improved efficacy against B. pseudomallei, which can survive within host cells where the local pH is acidic. In vitro analysis was performed using MICs, minimal bactericidal concentrations (MBCs), time-kill assays, persister cell assays, and macrophage assays. Finafloxacin showed increased bactericidal activity at pH 5 in comparison to pH 7 and ciprofloxacin at pH 5. In vivo studies in BALB/c mice included pharmacokinetic studies to inform an appropriate dosing regimen. Finafloxacin efficacy was evaluated in an inhalational murine model of melioidosis where antibiotic treatment was initiated at 6 or 24 h postchallenge and continued for 14 days, and mice were observed for 63 days. The survival of infected mice following 14 days of treatment was 80%, 60% or 0% for treatments initiated at 6 h and 60%, 30% or 0% for treatments initiated at 24 h for finafloxacin, co-trimoxazole, or ciprofloxacin, respectively. In summary, finafloxacin has increased bactericidal activity for B. pseudomallei under acidic conditions in vitro and improves survival in a murine model of melioidosis compared with those for ciprofloxacin. Furthermore, finafloxacin improves bacteriological clearance compared with that of co-trimoxazole, suggesting it may offer an effective postexposure prophylaxis against B. pseudomallei.


Global burden of Shigella infections: implications for vaccine development and implementation of control strategies.
Few studies provide data on the global morbidity and mortality caused by infection with Shigella spp.; such estimates are needed, however, to plan strategies of prevention and treatment. Here we report the results of a review of the literature published between 1966 and 1997 on Shigella infection. The data obtained permit calculation of the number of cases of Shigella infection and the associated mortality occurring worldwide each year, by age, and (as a proxy for disease severity) by clinical category, i.e. mild cases remaining at home, moderate cases requiring outpatient care, and severe cases demanding hospitalization. A sensitivity analysis was performed to estimate the high and low range of morbid and fatal cases in each category. Finally, the frequency distribution of Shigella infection, by serogroup and serotype and by region of the world, was determined. The annual number of Shigella episodes throughout the world was estimated to be 164.7 million, of which 163.2 million were in developing countries (with 1.1 million deaths) and 1.5 million in industrialized countries. A total of 69% of all episodes and 61% of all deaths attributable to shigellosis involved children under 5 years of age. The median percentages of isolates of S. flexneri, S. sonnei, S. boydii, and S. dysenteriae were, respectively, 60%, 15%, 6%, and 6% (30% of S. dysenteriae cases were type 1) in developing countries; and 16%, 77%, 2%, and 1% in industrialized countries. In developing countries, the predominant serotype of S. flexneri is 2a, followed by 1b, 3a, 4a, and 6. In industrialized countries, most isolates are S. flexneri 2a or other unspecified type 2 strains. Shigellosis, which continues to have an important global impact, cannot be adequately controlled with the existing prevention and treatment measures. Innovative strategies, including development of vaccines against the most common serotypes, could provide substantial benefits.


Tumor- and osteoclast-derived NRP2 in prostate cancer bone metastases.
Understanding the role of neuropilin 2 (NRP2) in prostate cancer cells as well as in the bone microenvironment is pivotal in the development of an effective targeted therapy for the treatment of prostate cancer bone metastasis. We observed a significant upregulation of NRP2 in prostate cancer cells metastasized to bone. Here, we report that targeting NRP2 in cancer cells can enhance taxane-based chemotherapy with a better therapeutic outcome in bone metastasis, implicating NRP2 as a promising therapeutic target. Since, osteoclasts present in the tumor microenvironment express NRP2, we have investigated the potential effect of targeting NRP2 in osteoclasts. Our results revealed NRP2 negatively regulates osteoclast differentiation and function in the presence of prostate cancer cells that promotes mixed bone lesions. Our study further delineated the molecular mechanisms by which NRP2 regulates osteoclast function. Interestingly, depletion of NRP2 in osteoclasts in vivo showed a decrease in the overall prostate tumor burden in the bone. These results therefore indicate that targeting NRP2 in prostate cancer cells as well as in the osteoclastic compartment can be beneficial in the treatment of prostate cancer bone metastasis.


When science develops outside state patronage: Dutch studies in Japan at the turn of the nineteenth century.
It is one of the peculiar features of the movement of translation of Western scientific treatises from Dutch into Japanese, known as Dutch learning (rangaku), that if first originated in Nagasaki with a group of Japanese interpreters. This group differed from the scholarly community of the capital, Edo, by both training and social status. This article shows how this difference contributes to explaining some of the particularities of rangaku in its initial phase. A case in point is Shizuki Tadao's introduction of Newtonian physics and astronomy. Yet, Sugita Genpaku, a major representative of the Edo scholarly community, gave an account of the beginning of Dutch learning that attempted to minimise, or even to erase, the contribution of the Nagasaki interpreters, who were dismissed as unscholarly. This attempt, too, is best understood in the light of the difference between the two communities.


Assessment of the impact of age and of blood-derived inflammatory markers in horses with colitis.
To determine the impact of age on survival in horses with colitis and to elucidate whether a lower type-1/type-2 cytokine ratio or an exaggerated inflammatory state contribute to reduced survival in aged horses. Part 1: Retrospective cohort analysis. Part 2: Analytic observational study. Part 1: One hundred twenty-four adult horses with colitis. Part 2: Twenty-nine adult horses with new diarrhea onset while hospitalized. Part 1: Patient signalment, select clinicopathological data, diagnoses, treatment, hospitalization length, and invoice were compared between survivors (n = 101) and nonsurvivors (n = 23). Only age and plasma transfusion retained statistical significance in the final multivariate outcome model, with 8.5 times lower odds of survival in transfused horses (95% confidence interval [CI], 2.6-27.2%). Additionally, the likelihood of nonsurvival increased by 11.8% (95% CI, 4-20.2%) for every year the horse aged (P = 0.002). Similarly, geriatric horses (≥20 years) were 15.2 times more likely to die than young-adults (2-12 years, P = 0.03), independent of financial investment, documented comorbidities, and duration of hospitalization. Part 2: Select cytokine analyses were performed on serum collected from hospitalized horses within 1 hour of diarrhea onset (T0) and 6 hours later. At T0, all recorded clinicopathological variables were comparable between geriatric and young-adult horses, suggesting a similar degree of systemic illness. The median concentration of type-2 cytokines interleukin-4 and interleukin-10, and type-1 cytokine interferon-γ did not differ between age groups. Inflammatory cytokines interleukin-6 and tumor necrosis factor-α were significantly higher in geriatric compared to young-adult horses at both sampling time points. Outcome of colitis was less favorable in aging horses and patients receiving a plasma transfusion. Although an exaggerated inflammatory state, based on increased interleukin-6 and tumor necrosis factor-α concentrations, in geriatric horses may contribute to reduced survival, a lower type-1/type-2 cytokines ratio was not identified in our geriatric population.


Effect of Music on Reducing Anxiety for Patients Undergoing Transrectal Ultrasound-Guided Prostate Biopsies: Randomized Prospective Trial.
The purpose of this study was to assess the impact of music on anxiety and perceived pain during transrectal ultrasound-guided prostate biopsy. Forty consecutive men with an elevated serum prostate specific antigen (PSA) level and/ or an abnormal digital rectal examination referred for transrectal ultrasound-guided prostate biopsy were recruited and allocated to a music (n = 20) or a non-music (n = 20) group. Anxiety was assessed prior to and after biopsy and pain was assessed after biopsy in each patient using visual analog scales (VAS) in the same setting, and group anxiety scores were compared. Patients in the music group experienced less anxiety (P = .046) during the procedure, but group pain scores were not significantly different. Music was found to decrease anxiety effectively during transrectal ultrasound-guided prostate biopsy.


Serial and subjective clustering on a verbal learning test (VLT) in children aged 5-15: the nature of subjective clustering.
This study investigated which strategies children aged 5-15 years (N = 408) employ while performing a multitrial free recall test of semantically unrelated words. Serial clustering (i.e., a relatively passive strategy) is an index of the sequential consistency of recall order. Subjective clustering (i.e., a more active strategy) is based on similar word groupings in successive trials. Previously, Meijs et al. (2009) found that the level of (serial and subjective) clustering increases with age. At all ages, the level of serial clustering correlates positively with the ability to recall information on VLT trials. However, subjective clustering is more predictive of VLT performance than serial clustering after ≥ 3 trials, but only in children aged 8+. Knowledge on how children organize words (based on, for example, sound or meaning) and how this relates to developmental stage is still lacking. This study revealed that the level of subjective clustering is primarily determined by the position of words in a VLT list. More specifically, primacy (i.e., recall of words 1-3 of the VLT list - whether recalled in the same order or reversed) and recency (i.e., recall of words 14-15) effects primarily determine level subjective organization over successive trials. Thus, older children still organize words based on the serial position of the VLT list and are much less likely to organize them based on any other feature of the words, for example, sound or meaning. This indicates that the most important information to be learned needs to be presented first or last, even in older children and even with repeated presentations.


On-line biofilm monitoring by "BIOX" electrochemical probe.
The innovative electrochemical monitoring probe (BIOX) recently developed to improve the antifouling treatments of cooling systems in industrial plants is presented. On the basis of the good results obtained from applications on marine sites, some research has been stated to validate this technique in biofilm growth and prevention of microbial corrosion in fresh and drinking waters.


Treatment of familial hemophagocytic lymphohistiocytosis with bone marrow transplantation from HLA genetically nonidentical donors.
Familial hemophagocytic lymphohistiocytosis (FHL) is a rare genetic disorder associated with the onset early in life of overwhelming activation of T lymphocytes and macrophages invariably leading to death. Allogeneic bone marrow transplantation (BMT) from an HLA-identical related donor is the treatment of choice in patients with this disease. However, fewer than 20% of patients have a disease-free HLA-identical sibling. BMT from HLA-nonidentical related donors has previously met with poor results, with graft rejection a major obstacle in all cases. We describe BMTs from HLA-nonidentical related donors (n = 13) and from a matched unrelated donor (n = 1) performed in two centers in 14 consecutive cases of FHL. Remission of disease was achieved before BMT in 10 patients. Marrow was T-cell-depleted to minimize graft-versus-host disease (GVHD). Antiadhesion antibodies specific for the alpha chain of the leukocyte function-associated antigen-1 (LFA-1, CD11a) and the CD2 molecules were infused pre-BMT and post-BMT to help prevent graft rejection, in addition to a conditioning regimen of busulfan (BU), cyclophosphamide (CP), and etoposide (VP16) or antithymocyte globulin (ATG). Sustained engraftment was obtained in 11 of 17 transplants (3 patients had 2 transplants) and disease-free survival in 9 patients with a follow-up period of 8 to 69 months (mean, 33). Acute GVHD greater than stage I was not observed, and 1 patient had mild cutaneous chronic GVHD that resolved. Toxicity due to the BMT procedure was low. Results obtained using this protocol are promising in terms of engraftment and event-free survival within the limitations of the small sample. We conclude that an immunologic approach in terms of drugs used to obtain disease remission and a conditioning regimen that includes antiadhesion molecules in T-cell-depleted BMT from HLA genetically nonidentical donors is an alternative treatment that warrants further study in FHL patients who lack a suitable HLA genetically identical donor.


PPARγ controls pregnancy outcome through activation of EG-VEGF: new insights into the mechanism of placental development.
PPARγ-deficient mice die at E9.5 due to placental abnormalities. The mechanism by which this occurs is unknown. We demonstrated that the new endocrine factor EG-VEGF controls the same processes as those described for PPARγ, suggesting potential regulation of EG-VEGF by PPARγ. EG-VEGF exerts its functions via prokineticin receptor 1 (PROKR1) and 2 (PROKR2). This study sought to investigate whether EG-VEGF mediates part of PPARγ effects on placental development. Three approaches were used: 1) in vitro, using human primary isolated cytotrophoblasts and the extravillous trophoblast cell line (HTR-8/SVneo); 2) ex vivo, using human placental explants (n = 46 placentas); and 3) in vivo, using gravid wild-type PPARγ(+/-) and PPARγ(-/-) mice. Major processes of placental development that are known to be controlled by PPARγ, such as trophoblast proliferation, migration, and invasion, were assessed in the absence or presence of PROKR1 and PROKR2 antagonists. In both human trophoblast cell and placental explants, we demonstrated that rosiglitazone, a PPARγ agonist, 1) increased EG-VEGF secretion, 2) increased EG-VEGF and its receptors mRNA and protein expression, 3) increased placental vascularization via PROKR1 and PROKR2, and 4) inhibited trophoblast migration and invasion via PROKR2. In the PPARγ(-/-) mouse placentas, EG-VEGF levels were significantly decreased, supporting an in vivo control of EG-VEGF/PROKRs system during pregnancy. The present data reveal EG-VEGF as a new mediator of PPARγ effects during pregnancy and bring new insights into the fine mechanism of trophoblast invasion.


[Comparison of boys' and girls' families for actor and partner effect of stress, depression and parent-adolescent communication on middle school students' suicidal ideation: triadic data analysis].
This study was done to compare families of boys or of girls for actor and partner effect of stress, depression and parent-adolescent communication as perceived by mother, father and adolescent on adolescents' suicidal ideation. Participants were 183 families (104 boys' families, 79 girls' families) who met eligibility criteria. All measures were self-administered. Data were analyzed using SPSS 18.0 and AMOS 18.0 program. In boys' families, boys' depression and communication with father showed actor effect on boys' suicidal ideation. Boys' stress showed indirect effect on boys' suicidal ideation through communication with father and boys' depression. Mothers' depression showed indirect partner effect on boys' suicidal ideation through boys' depression. In families of girls, girls' depression and stress showed actor effects on girls' suicidal ideation. Girls' communication with mother showed indirect effects through girls' depression. Also girls' stress showed indirect effect through girls' depression. Stress in mothers and/or fathers showed partner effect on girls' suicidal ideation. To intervene in adolescents' suicidal ideation and promote adolescents' mental health, programs should be developed differently according to gender and based on parent's psychological states.


Nasal toxicity, carcinogenicity, and olfactory uptake of metals.
Occupational exposures to inhalation of certain metal dusts or aerosols can cause loss of olfactory acuity, atrophy of the nasal mucosa, mucosal ulcers, perforated nasal septum, or sinonasal cancer. Anosmia and hyposmia have been observed in workers exposed to Ni- or Cd-containing dusts in alkaline battery factories, nickel refineries, and cadmium industries. Ulcers of the nasal mucosa and perforated nasal septum have been reported in workers exposed to Cr(VI) in chromate production and chrome plating, or to As(III) in arsenic smelters. Atrophy of the olfactory epithelium has been observed in rodents following inhalation of NiSO4 or alphaNi3S2. Cancers of the nose and nasal sinuses have been reported in workers exposed to Ni compounds in nickel refining, cutlery factories, and alkaline battery manufacture, or to Cr(VI) in chromate production and chrome plating. In animals, several metals (eg, Al, Cd, Co, Hg, Mn, Ni, Zn) have been shown to pass via olfactory receptor neurons from the nasal lumen through the cribriform plate to the olfactory bulb. Some metals (eg, Mn, Ni, Zn) can cross synapses in the olfactory bulb and migrate via secondary olfactory neurons to distant nuclei of the brain. After nasal instillation of a metal-containing solution, transport of the metal via olfactory axons can occur rapidly, within hours or a few days (eg, Mn), or slowly over days or weeks (eg, Ni). The olfactory bulb tends to accumulate certain metals (eg, Al, Bi, Cu, Mn, Zn) with greater avidity than other regions of the brain. The molecular mechanisms responsible for metal translocation in olfactory neurons and deposition in the olfactory bulb are unclear, but complexation by metal-binding molecules such as carnosine (beta-alanyl-L-histidine) may be involved.


Effect of blood tamoxifen concentrations on surrogate biomarkers in a trial of dose reduction in healthy women.
Tamoxifen administered at 20 mg/d has been shown to decrease breast cancer incidence in at-risk women by 50%, but toxicity may limit its broad use, particularly in postmenopausal women. Because toxicity may be dose-dependent, we studied the biologic activity of low concentrations of tamoxifen to determine the plausibility of a dose reduction. We measured the blood concentrations of tamoxifen and its main metabolites in a dose titration study in 105 healthy women (placebo, tamoxifen 10 mg on alternate days, tamoxifen 10 mg/d, and tamoxifen 20 mg/d). Drug levels measured after 2 months of treatment were correlated with the changes in surrogate biomarkers of different diseases, including lipid profile, blood cell count, fibrinogen, antithrombin III, osteocalcin, and insulin-like growth factor I, a promising surrogate biomarker of breast cancer. The means (+/- SD) for tamoxifen and N-desmethyltamoxifen (metabolite X) concentrations (ng/mL) were dose-related, being, respectively, 0 and 0 with placebo, 26.8 +/- 15.1 and 43.7 +/- 22.5 with 10 mg every other day, 51.2 +/- 24.1 and 90.7 +/- 48.0 with 10 mg/d, and 136.0 +/- 52.7 and 230.6 +/- 75.0 with 20 mg/d of tamoxifen. At variance, the biomarker changes were of comparable magnitude at any drug concentration except for platelet count and triglycerides levels, the latter showing a trend to an increase with increasing tamoxifen concentrations. An 80% reduction in blood concentrations does not seem to affect the activity of tamoxifen on biomarkers of cardiovascular or breast cancer risk and may in fact have a more favorable safety profile. Additional studies are warranted to determine the most appropriate dose of this agent.


Rosmarinic Acid Potently Detoxifies Amylin Amyloid and Ameliorates Diabetic Pathology in a Transgenic Rat Model of Type 2 Diabetes.
Protein aggregation is associated with a large number of human protein-misfolding diseases, yet FDA-approved drugs are currently not available. Amylin amyloid and plaque depositions in the pancreas are hallmark features of type 2 diabetes. Moreover, these amyloid deposits are implicated in the pathogenesis of diabetic complications such as neurodegeneration. We recently discovered that catechols and redox-related quinones/anthraquinones represent a broad class of protein aggregation inhibitors. Further screening of a targeted library of natural compounds in complementary medicine that were enriched with catechol-containing compounds identified rosmarinic acid (RA) as a potent inhibitor of amylin aggregation (estimated inhibitory concentration IC50 = 200-300 nM). Structure-function relationship analysis of RA showed the additive effects of the two catechol-containing components of the RA molecule. We further showed that RA does not reverse fibrillation back to monomeric amylin but rather lead to nontoxic, remodeled protein aggregates. RA has significant ex vivo efficacy in reducing human amylin oligomer levels in HIP rat sera as well as in sera from diabetic patients. In vivo efficacy studies of RA treatment with the diabetic HIP rat model demonstrated significant reduction in amyloid islet deposition and strong mitigation of diabetic pathology. Our work provides new in vitro molecular mechanisms and in vivo efficacy insights for a model nutraceutical agent against type 2 diabetes and other aging-related protein-misfolding diseases.


Enhanced dissolution of silver nanoparticles in a physical mixture with platinum nanoparticles based on the sacrificial anode effect.
A strategy to reduce implant-related infections is the inhibition of the initial bacterial implant colonization by biomaterials containing silver (Ag). The antimicrobial efficacy of such biomaterials can be increased by surface enhancement (nanosilver) or by creating a sacrificial anode system for Ag. Such a system will lead to an electrochemically driven enhanced Ag ion release due to the presence of a more noble metal. Here we combined the enlarged surface of nanoparticles (NP) with a possible sacrificial anode effect for Ag induced by the presence of the electrochemically more noble platinum (Pt) in physical mixtures of Ag NP and Pt NP dispersions. These Ag NP/Pt NP mixtures were compared to the same amounts of pure Ag NP in terms of cell biological responses, i.e. the antimicrobial activity against Staphylococcus aureus and Escherichia coli as well as the viability of human mesenchymal stem cells (hMSC). In addition, Ag NP was analyzed by ultraviolet-visible (UV-vis) spectroscopy, cyclic voltammetry, and atomic absorption spectroscopy. It was found that the dissolution rate of Ag NP was enhanced in the presence of Pt NP within the physical mixture compared to a dispersion of pure Ag NP. Dissolution experiments revealed a fourfold increased Ag ion release from physical mixtures due to enhanced electrochemical activity, which resulted in a significantly increased toxicity towards both bacteria and hMSC. Thus, our results provide evidence for an underlying sacrificial anode mechanism induced by the presence of Pt NP within physical mixtures with Ag NP. Such physical mixtures have a high potential for various applications, for example as antimicrobial implant coatings in the biomedicine or as bactericidal systems for water and surface purification in the technical area.


Optimization and In Vivo Profiling of a Refined Rat Model of Walker 256 Breast Cancer Cell-Induced Bone Pain Using Behavioral, Radiological, Histological, Immunohistochemical and Pharmacological Methods.
In the majority of patients with advanced breast cancer, there is metastatic spread to bones resulting in pain. Clinically available drug treatments for alleviation of breast cancer-induced bone pain (BCIBP) often produce inadequate pain relief due to dose-limiting side-effects. A major impediment to the discovery of novel well-tolerated analgesic agents for the relief of pain due to bony metastases is the fact that most cancer-induced bone pain models in rodents relied on the systemic injection of cancer cells, causing widespread formation of cancer metastases and poor general animal health. Herein, we have established an optimized, clinically relevant Wistar Han female rat model of breast cancer induced bone pain which was characterized using behavioral assessments, radiology, histology, immunohistochemistry and pharmacological methods. In this model that is based on unilateral intra-tibial injection (ITI) of Walker 256 carcinoma cells, animals maintained good health for at least 66 days post-ITI. The temporal development of hindpaw hypersensitivity depended on the initial number of Walker 256 cells inoculated in the tibiae. Hindpaw hypersensitivity resolved after approximately 25 days, in the continued presence of bone tumors as evidenced by ex vivo histology, micro-computed tomography scans and immunohistochemical assessments of tibiae. A possible role for the endogenous opioid system as an internal factor mediating the self-resolving nature of BCIBP was identified based upon the observation that naloxone, a non-selective opioid antagonist, caused the re-emergence of hindpaw hypersensitivity. Bolus dose injections of morphine, gabapentin, amitriptyline and meloxicam all alleviated hindpaw hypersensitivity in a dose-dependent manner. This is a first systematic pharmacological profiling of this model by testing standard analgesic drugs from four important diverse classes, which are used to treat cancer induced bone pain in the clinical setting. Our refined rat model more closely mimics the pathophysiology of this condition in humans and hence is well-suited for probing the mechanisms underpinning breast cancer induced bone pain. In addition, the model may be suitable for efficacy profiling of new molecules from drug discovery programs with potential to be developed as novel agents for alleviation of intractable pain associated with disseminated breast cancer induced bony metastases.


Follow-up treatment for osteoporosis after fracture.
Studies of the management of osteoporosis in older women who have had hip or wrist fractures have found underdiagnosis and undertreatment of the disease. Few such studies have been conducted in the United States, however, and most studies have been confined to a subset of the treatments currently available to treat osteoporosis. Mail surveys were sent to 381 women between 50 and 84 years of age who had been treated for a hip or wrist fracture at a large northeast US teaching hospital between October 1, 1998, and September 30, 2000. These surveys included questions about osteoporosis risk factors and physician treatment both before and after the index fracture. Of 381 surveys mailed, 70 were returned because of an invalid address or by a relative because a patient was deceased. Of the remaining 311 surveys, 147 completed responses were received. Fifty-two percent of respondents reported having received either a prescription or a recommendation for a nonprescription medication used to treat osteoporosis before the fracture. After fracture, 60% of subjects were advised to take any osteoporosis medication, and 42% of were advised to take a prescription medication. Of women reporting no treatment advice before fracture, 33% reported treatment after. Twenty-four percent of patients reported a change in treatment after fracture versus before. No significant differences in treatment were found according to fracture history, maternal history of fracture, or maternal history of osteoporosis. Both prescription and nonprescription treatment prevalence after fracture were lower than expected, and there was only a small change in reported treatment prevalence after fracture versus before. There was also little difference in treatment prevalence based on risk factors for osteoporosis or osteoporotic fractures. A sizeable opportunity exists for intervention to reduce the risk of osteoporotic fractures for patients who have a history of fracture.


The requirements for a high level of transposition of bacteriophage Mu.
To analyse the functional and structural requirements of Mu transposition a special mini-Mu transposon was constructed. This mini-Mu, situated on a derivative of pBR322, has an easily selectable marker gene conferring chloramphenicol resistance (Cam) cloned between the ends of the mini-Mu. The genes required for transposition are cloned outside the ends and are under control of the PL promoter of bacteriophage lambda. To obtain a high frequency of transposition the A and B products are required. Under B- conditions both cointegrate formation and simple insertions are strongly reduced. The role of B in the transposition process is unknown and is discussed. Deletion analysis of the ends of the mini-Mu has revealed the existence of multiple sites required for transposition. The same sites were found to be strong A-binding sites. In addition two weak A-binding sites were found. Point mutations introduced into these last sites show that they are at least as important for transposition as the strong binding sites. The organization of the binding sites is asymmetric in Mu. At the left end two binding sites are situated more than 100 bp away from the end. These affect specifically the frequency of transposition. The distance between these two sites and the site close to the left end is extremely critical. At the right end also three A-binding sites are found, but only two seem to be important for transposition. Transposition with low frequency can be obtained with a mini-Mu with only one end. Secondary att sites are selected which all show some homology with an A-binding site. An absolute requirement for a 5'T at the artificial end is observed. Based on these results a simple scheme for the evolution of a transposon is discussed.


Clinical Benefit of Tyrosine Kinase Inhibitors in Advanced Lung Cancer with EGFR-G719A and Other Uncommon EGFR Mutations.
The optimal management of advanced non-small cell lung cancer (NSCLC) with noncanonical epidermal growth factor receptor (EGFR) mutations (i.e., exon 19 deletion and exon 21 L858R) is constrained by the heterogeneous behavior of individual uncommon mutations and limited prospective clinical data in this setting. Despite encouraging results with osimertinib from a recently published phase II trial from South Korea, afatinib remains the only currently approved drug for patients with tumors harboring uncommon EGFR mutations (i.e., S768I, L861Q, and/or G719X). When used at the standard dose of 40 mg daily, afatinib is associated with significant rates of treatment-related adverse events, leading to frequent dose reductions and treatment discontinuations. We report a case of a woman with advanced NSCLC harboring EGFR-G719A mutation treated with afatinib (at an off-label pulse dose strategy that merits further evaluation in prospective studies) with sustained partial response for 20 months with manageable expected toxicities. Subsequent disease progression was mediated by off-target pan-EGFR inhibitor (including osimertinib)-resistant KRAS mutation and not by acquisition of EGFR-T790M. We further present the current state of evidence in the literature behind use of first-, second-, and third-generation tyrosine kinase inhibitors and summarize the evolving spectrum of activity ascribed to osimertinib (and newer EGFR inhibitors with a more favorable therapeutic window and intracranial penetration) in this population of patients with advanced NSCLC and uncommon EGFR mutations. KEY POINTS: Uncommon EGFR mutations characterize a heterogeneous group of patients with advanced non-small cell lung cancer (NSCLC). Afatinib is the only currently U.S. Food and Drug Administration-approved drug for management of advanced NSCLC with uncommon EGFR mutations (S768I, L861Q, and/or G719X). Afatinib treatment at 40 mg daily is associated with high rates of adverse events and dose reductions; alternative strategies including pulse intermittent dosing should be evaluated prospectively. Osimertinib (with favorable safety profile and intracranial penetration) has shown promising results in this population in a phase II trial from South Korea; additional trials are ongoing.


Neural dynamics between anterior insular cortex and right supramarginal gyrus dissociate genuine affect sharing from perceptual saliency of pretended pain.
Empathy for pain engages both shared affective responses and self-other distinction. In this study, we addressed the highly debated question of whether neural responses previously linked to affect sharing could result from the perception of salient affective displays. Moreover, we investigated how the brain network involved in affect sharing and self-other distinction underpinned our response to a pain that is either perceived as genuine or pretended (while in fact both were acted for reasons of experimental control). We found stronger activations in regions associated with affect sharing (anterior insula [aIns] and anterior mid-cingulate cortex) as well as with affective self-other distinction (right supramarginal gyrus [rSMG]), in participants watching video clips of genuine vs. pretended facial expressions of pain. Using dynamic causal modeling, we then assessed the neural dynamics between the right aIns and rSMG in these two conditions. This revealed a reduced inhibitory effect on the aIns to rSMG connection for genuine pain compared to pretended pain. For genuine pain only, brain-to-behavior regression analyses highlighted a linkage between this inhibitory effect on the one hand, and pain ratings as well as empathic traits on the other. These findings imply that if the pain of others is genuine and thus calls for an appropriate empathic response, neural responses in the aIns indeed seem related to affect sharing and self-other distinction is engaged to avoid empathic over-arousal. In contrast, if others merely pretend to be in pain, the perceptual salience of their painful expression results in neural responses that are down-regulated to avoid inappropriate affect sharing and social support.


Digestibility index and factors affecting rate of starch digestion in vitro in conventional food preparation.
The rate of starch hydrolysis in ten cereal-based food preparations was studied using an in vitro dialysis system. The foods were incubated with human saliva and porcine pancreatin. The sugars released after 3 h digestion were expressed as digestibility index (DI), the percentage starch digested was determined and correlated with the degree of gelatinization (DG). Granule morphology was also investigated and related with starch availability for hydrolysis. Significant differences were observed in the in vitro starch digestibility of the 10 foods (P < 0.05). The DI ranged from 53 for chapathi to 78 for rice flakes. DI was inversely related to the protein (r = -0.79, P < 0.01), fat (r = -0.63, P < 0.05) and energy (r = -0.61, P < 0.01). Percent starch digested was inversely related to the insoluble (r = -0.49, P < 0.05) and total dietary fiber (r = -0.63, P < 0.01) content of the foods. The SEM results provided a better understanding of granular morphology on cooking and the effect of protein on limiting DG. The results suggest that carbohydrate foods of potential use in the therapeutic diets may be identified by their in vitro digestion characteristics.


Mechanisms of amphotericin B-induced decrease in glomerular filtration rate in rats.
Mechanisms responsible for the reductions in renal blood flow (RBF) and glomerular filtration rate (GFR) in response to acute infusions of amphotericin B were investigated in vivo in rats. The influence of salt status and the roles of adenosine, cyclic AMP, and calcium influx were examined. Amphotericin B was infused into the renal artery in seven groups of rats at 0.025 mg/kg of body weight per min for 15 min. RBF and GFR were measured over 15 min before, during, and after the infusion. Control rats were maintained on a normal salt diet; a second group of rats received a salt-depleted diet, and a third group received a high-salt intake. Four other groups were kept on a normal diet and received theophylline (0.5 mumol/kg/min into the renal artery, intra-arterially [i.a.]), dibutyryl cyclic AMP (85 micrograms/min, i.a.), the 5'-nucleotidase inhibitor adenosine alpha,beta-methylene diphosphate (4 mg/kg, intramuscularly), or diltiazem (20 micrograms/kg/min, i.a.). Control rats had a prompt 50% decrease in RBF in response to amphotericin B. This was sustained over the 15-min infusion period and was accompanied by a decrease in creatinine clearance (CLCR) (from 0.83 +/- 0.08 to 0.40 +/- 0.09 ml/min; P less than 0.05). On stopping the infusion, RBF returned quickly to baseline but CLCR continued to decrease further (to 0.35 +/- 0.07 ml/min; P less than 0.05). Salt loading, theophylline, and diltiazem administration prevented the decreases in both RBF and CLCR. Both RBF and CLCR responses in the remaining groups were not significantly different from those in controls. The results of this study reveal a protective effect of salt loading and theophylline against amphotericin B nephrotoxicity in the rat but deny a role for adenosine in mediating these effects. They further suggest that theophylline inhibits the acute responses by a mechanism unrelated to either adenosine receptor blockade or phosphodiesterase inhibition and that calcium influx into the cells is probably responsible for the acute changes in RBF and GFR in response to amphotericin B.


The importance of intestinal endotoxins in liver disease.
The development and wider use of the Limulus amoebocyte lysate assay (LAL) for the detection of endotoxin, has led to renewed interest in the link between gut-derived endotoxin, liver injury and the extra-hepatic manifestations of clinical liver disease. The concept that endotoxemia may occur and be harmful in clinical states without the concomitant presence of Gram-negative bacteria is a relatively recent one and has been most intensively studied in the context of liver injury. Since the liver stands between the gut and the systemic circulation, it has been postulated that failure to detoxify endotoxins absorbed into the portal circulation after hepatic injury might lead to further liver damage and escape of this toxic material into the general circulation. The present review will update the status of this hypothesis, discuss mechanisms proposed for the damage and present evidence for the association in animal models. Finally, studies both positive and negative, will be cited that have used the LAL to detect endotoxemia and its consequences in human liver disease.


Human immunodeficiency virus (HIV) and stroke: targets for intervention.
Human immunodeficiency virus (HIV) infection causes stroke through several mechanisms. Stroke results from opportunistic infection and neoplasia, HIV induced cardiac disease, HIV associated cerebral vasculopathy, and perhaps of HIV induced facilitation of some forms of systemic vasculitis and prothrombotic haematological conditions. HIV causes more ischaemic stroke than cerebral haemorrhage. Although stroke is currently a relatively infrequent manifestation of HIV infection, the incidence of stroke in HIV infected individuals is likely to increase with current combination antiretroviral therapy. HIV infection per se induces endothelial activation and dyslipidaemia, predisposing to accelerated atherosclerosis. Antiretroviral therapy, which increases life expectancy and therefore inherently increases ischaemic stroke risk with advancing age and length of exposure to traditional risk factors, also causes pro-atherosclerotic metabolic and endothelial dysfunction. Antiretroviral induced vascular dysfunction together with pre-existing HIV induced vascular disease has the potential to increase atherosclerotic causes of ischaemic stroke. New antiretroviral agents should ideally eradicate the human immunodeficiency virus thereby reducing vascular risk and HIV related causes of stroke without inducing metabolic or endothelial dysfunction. Future studies of vascular disease in HIV infected individuals, particularly studies investigating the impact of current and future antiretroviral agents, should ideally assess stroke as a specific outcome, and provide data by pathological stroke type and ischaemic stroke subtype, to clarify the mechanisms of stroke and guide the approach to treatment and prevention of stroke.


Single-dose pharmacokinetics and tolerance of a cholesteryl sulfate complex of amphotericin B administered to healthy volunteers.
Twenty-three healthy volunteer subjects received a single dose of amphotericin B colloidal dispersion or placebo (4:2) in a double-blind, randomized, dose-escalating design. Doses ranged from 0.25 to 1.5 mg/kg of body weight. The medication was administered via intravenous infusion at a rate of 0.5 mg/kg/h. Plasma amphotericin B concentrations increased with increasing doses, resulting in a linear increase in the amphotericin B area under the curve. Concentrations in plasma decreased rapidly upon discontinuation of the infusion, indicating rapid tissue distribution. A log-linear biexponential elimination phase was observed. A three-compartment open model was used to describe the distribution and elimination of amphotericin B. The mean terminal elimination half-life ranged from 86 h at the 0.25-mg/kg dose level to 244 and 235 h at the 1.0- and 1.5-mg/kg dose levels, respectively. Mean total body clearance ranged from 219 to 284 ml/kg/h. The volume of distribution increased with dose, from 3.37 liter/kg at the 0.25-mg/kg dose to 7.92 liter/kg at the 1.5-mg/kg dose. At the lowest dose level, 0.25 mg/kg, the medication was generally well tolerated. Progressive increases in the dose led to increasing side effects. At the 1.5-mg/kg dose level, 50% of the patients on active medication experienced nausea, vomiting, and chills. Physical examinations, ophthalmologic examinations, and clinical laboratory parameters remained within normal limits compared with those obtained during prestudy examinations.


The relevance of a multidomain geriatric assessment in older patients with heart failure.
Physical frailty screening is more commonly performed at outpatient heart failure (HF) clinics. However, this does not incorporate other common geriatric domains. This study assesses whether a multidomain geriatric assessment, in comparison with HF severity or physical frailty, is associated with short-term adverse outcomes. This is a prospective cohort study of 197 patients with HF (mean age 78, 44% female) attending outpatient HF clinics. HF severity was assessed with New York Heart Association class (I-II versus III-IV) and N-terminal pro b-type natriuretic peptide levels. Physical frailty was assessed with the Fried frailty criteria (not frail, pre-frail, and frail). The following geriatric domains were assessed: physical function, nutrition, polypharmacy, cognition, and dependency in activities of daily living. Logistic regression analyses adjusted for age, sex, diabetes and kidney function assessed 3 month risk of adverse health outcomes (emergency department visits, hospital admissions, and/or death) according to HF severity, physical frailty, and number of affected domains. Number (%) of patients with HF with no, 1, 2, and ≥3 domains affected were 36 (18%), 61 (31%), 58 (29%), and 42 (21%). Seventy-four adverse outcomes were experienced in 50 patients at follow-up. Severity of HF and physical frailty were not significantly associated with an increased risk of adverse health outcomes. However, increasing number of affected domains were significantly associated with an increased risk of adverse outcomes. Compared with no domains affected, odds ratios (95% confidence interval) for 1, 2, and ≥3 domains were 1.8 (0.5-6.5), 4.5 (1.3-15.4), and 7.2 (2.0-26.3) (P-trend <0.01). Further adjustment for HF severity and frailty status slightly attenuated the effect estimates (P-trend 0.02). Having limitations in multiple domains appears more strongly associated with short-term adverse outcomes than HF severity and physical frailty. This may illustrate the potential added value of a multidomain geriatric assessment in the evaluation and treatment of patients with HF with respect to relevant short-term health outcomes.


jMOTU and Taxonerator: turning DNA Barcode sequences into annotated operational taxonomic units.
DNA barcoding and other DNA sequence-based techniques for investigating and estimating biodiversity require explicit methods for associating individual sequences with taxa, as it is at the taxon level that biodiversity is assessed. For many projects, the bioinformatic analyses required pose problems for laboratories whose prime expertise is not in bioinformatics. User-friendly tools are required for both clustering sequences into molecular operational taxonomic units (MOTU) and for associating these MOTU with known organismal taxonomies. Here we present jMOTU, a Java program for the analysis of DNA barcode datasets that uses an explicit, determinate algorithm to define MOTU. We demonstrate its usefulness for both individual specimen-based Sanger sequencing surveys and bulk-environment metagenetic surveys using long-read next-generation sequencing data. jMOTU is driven through a graphical user interface, and can analyse tens of thousands of sequences in a short time on a desktop computer. A companion program, Taxonerator, that adds traditional taxonomic annotation to MOTU, is also presented. Clustering and taxonomic annotation data are stored in a relational database, and are thus amenable to subsequent data mining and web presentation. jMOTU efficiently and robustly identifies the molecular taxa present in survey datasets, and Taxonerator decorates the MOTU with putative identifications. jMOTU and Taxonerator are freely available from http://www.nematodes.org/.


Synergistic activity of everolimus and 5-aza-2'-deoxycytidine in medullary thyroid carcinoma cell lines.
Medullary thyroid cancer (MTC) is a tumor highly resistant to chemo- and radiotherapy. Drug resistance can be induced by epigenetic changes such as aberrant DNA methylation. To overcome drug resistance, we explored a promising approach based on the use of 5-aza-2'-deoxycytidine (AZA), a demethylating agent, in combination with the mTOR inhibitor everolimus in MTC cells (MZ-CRC-1 and TT). This combined treatment showed a strong synergistic antiproliferative activity through the induction of apoptosis. The effect of everolimus and/or AZA on genome-wide expression profiling was evaluated by Illumina BeadChip in MZ-CRC-1 cells. An innovative bioinformatic pipeline identified four potential molecular pathways implicated in the synergy between AZA and everolimus: PI3K-Akt signaling, the neurotrophin pathway, ECM/receptor interaction, and focal adhesion. Among these, the neurotrophin signaling pathway was most directly involved in apoptosis, through the overexpression of NGFR and Bax genes. The increased expression of genes involved in the NGFR-MAPK10-TP53-Bax/Bcl2 pathway during incubation with AZA plus everolimus was validated by western blotting in MZ-CRC-1 cells. Interestingly, addition of a neutralizing anti-NGFR antibody inhibited the synergistic cytotoxic activity between AZA and everolimus. These results open a new therapeutic scenario for MTC and potentially other neuroendocrine tumors, where therapy with mTOR inhibitors is currently approved.


Safflower polysaccharide inhibits the development of tongue squamous cell carcinoma.
Safflower polysaccharide (SPS) is one of the most important active components of safflower (Carthamus tinctorius L.), which has been confirmed to have the immune-regulatory function and antitumor effect. This study aimed to explore the effects of safflower polysaccharide (SPS) on tongue squamous cell carcinoma (TSCC). HN-6 cells were treated with 5 μg/mL cisplatin and various concentrations of SPS (0, 0.02, 0.04, 0.08, 0.16, 0.32, 0.64, and 1.28 mg/mL), and cell proliferation was measured. After treatment with 5 μg/mL cisplatin and 0.64 mg/mL SPS, the induction of apoptosis and the protein and mRNA expression of Bax, Bcl-2, COX-2, and cleaved caspase-3 in HN-6 cells were quantified. In addition, HN-6 cells were implanted into mice to establish an in vivo tumor xenograft model. Animals were randomly assigned to three groups: SPS treatment, cisplatin treatment, and the model group (no treatment). The body weight, tumor volume, and tumor weight were measured, and the expression of the above molecules was determined. SPS treatment (0.02-0.64 mg/mL) for 24-72 h inhibited HN-6 cell proliferation. In addition, 0.64 mg/mL SFP markedly induced apoptosis in HN-6 cells and arrested the cell cycle at the G0/G1 phase. Compared with the control group, the expression of Bcl-2 and COX-2 was markedly reduced by SPS treatment, whereas the expression of Bax and cleaved caspase-3 was increased. Moreover, SPS significantly inhibited the growth of the tumor xenograft, with similar changes in the expression of Bcl-2, COX-2, Bax, and cleaved caspase-3 in the tumor xenograft to the in vitro analysis. Our results indicated that SPS may inhibit TSCC development through regulation of Bcl-2, COX-2, Bax, and cleaved caspase-3 expression.


TADA: Taxonomy-Aware Dataset Aggregator.
The profusion of sequenced genomes across the bacterial and archeal domains offers unprecedented possibilities for phylogenetic and comparative genomic analyses. In general, phylogenetic reconstruction is improved by the use of more data. However, including all available data is (i) not computationally tractable, and (ii) prone to biases, as the abundance of genomes is very unequally distributed over the biological diversity. Thus, in most cases, subsampling taxa to build a phylogeny is necessary. Currently, though, there is no available software to perform that handily. Here we present TADA, a taxonomic-aware dataset selection workflow that allows sampling across user-defined portions of the prokaryotic diversity with variable granularity, while setting constraints on genome quality and balance between branches. TADA is implemented as a snakemake workflow and is freely available at https://github.com/emilhaegglund/TADA. Supplementary data are available at Bioinformatics online.


Comparing Smoking Cessation Outcomes in Nurse-Led and Physician-Led Primary Care Visits.
Smoking is a significant public health concern in the United States, yet 50% of patients do not receive recommended tobacco use screening and counseling. This project compared smoking cessation rates in newly reimbursable nurse-led wellness visits with rates in physician-led visits. Although the findings were not statistically significant, they suggested that smoking cessation is at least equivalent in patients who attend nurse-led visits compared with physician-led visits and may be higher.


Identification and management of young infants with possible serious bacterial infection where referral was not feasible in rural Lucknow district of Uttar Pradesh, India: An implementation research.
Based on World Health Organization guidelines, Government of India recommended management of possible serious bacterial infection (PSBI) in young infants up to two months of age on an outpatient basis where referral is not feasible. We implemented the guideline in program setting to increase access to treatment with high treatment success and low resultant mortality. Implementation research was conducted in four rural blocks of Lucknow district in Uttar Pradesh, India. It included policy dialogues with the central and state government and district level officials. A Technical Support Unit was established. Thereafter, capacity building across all cadres of health workers in the implementation area was done for strengthening of home based newborn care (HBNC) program, skills enhancement for identification and management of PSBI, logistics management to ensure availability of necessary supplies, monitoring and evaluation as well as providing feedback. Data was collected by the research team. From June 2017 to February 2019 there were 24,448 live births in a population of 856106. We identified 1302 infants, aged 0-59 days, with any sign of PSBI leading to a coverage of 53% (1302/2445), assuming an incidence of 10%. However, in the establishment phase the coverage was 33%, while it was 85% in the implementation phase. Accredited social health activists (ASHAs) identified 81.2% (1058/1302) cases while rest were identified by families. ASHAs increased home visits within first 7 days of life in home based newborn care program from 74.3% (2781/3738) to 89.0% (3128/3513) and detection of cases of PSBI from 1.6% (45/2781) to 8.7% (275/3128) in the first and last quarter of the project, respectively. Of these 18.7% (244/1302) refused referral to government health system and 6.7% (88/1302) were treated in a hospital. Among cases of PSBI, there were 13.3% (173/1302) cases of fast breathing in young infant aged 7-59 days in whom referral was not needed. Of these 147 were treated by oral amoxicillin and 95.2% (140/147) were cured. Among those who needed referral, simplified treatment was given when referral was refused. There were 2.9% (37/1302) cases of fast breathing at ages of 0-6 days of which 34 were treated by simplified treatment with100% (34/34) cured;66.5% (866/1302) were cases of clinical severe infection of which 685 treated by simplified treatment with94.2% (645/685)cured and 09 died;17.3% (226/1302) cases of critical illness of which 93 were treated by simplified treatment, as a last resort, 72% (67/93) cured and 16 died. Among 255 cases who either did not seek formal treatment or sought it at private facilities, 96 died. Simplified treatment for PSBI is feasible in public program settings in northern India with good cure rates. It required system strengthening and supportive supervision.


Femoral varus-derotation osteotomy in spastic cerebral palsy.
Twenty children (twenty hips) with spastic cerebral palsy underwent femoral varus-derotation osteotomy for which the principal indication was inadequate coverage of the femoral head. Other infrequent indications included pain, valgus angulation of the femoral neck, and dislocation of the hip. The age at surgery ranged from four to fifteen years. Each child had muscle releases before the osteotomy was done. At follow-up, at least seven years later, the femoral head in twenty hips was well centered. Four hips remained subluxated but were less subluxated than before the osteotomy. One hip remained dislocated. The center-edge angle averaged -8 degrees preoperatively and +17 degrees at follow-up. The neck-shaft angle averaged 155 degrees preoperatively, 114 degrees immediately post-operatively, and 125 degrees at follow-up. Femoral varus-derotation osteotomy, when combined with muscle releases, can allow children with spastic cerebral palsy to maintain their ambulatory status and may decrease pain about the hip.


Regulation of rhodopsin-eGFP distribution in transgenic xenopus rod outer segments by light.
The rod outer segment (OS), comprised of tightly stacked disk membranes packed with rhodopsin, is in a dynamic equilibrium governed by a diurnal rhythm with newly synthesized membrane inserted at the OS base balancing membrane loss from the distal tip via disk shedding. Using transgenic Xenopus and live cell confocal imaging, we found OS axial variation of fluorescence intensity in cells expressing a fluorescently tagged rhodopsin transgene. There was a light synchronized fluctuation in intensity, with higher intensity in disks formed at night and lower intensity for those formed during the day. This fluctuation was absent in constant light or dark conditions. There was also a slow modulation of the overall expression level that was not synchronized with the lighting cycle or between cells in the same retina. The axial variations of other membrane-associated fluorescent proteins, eGFP-containing two geranylgeranyl acceptor sites and eGFP fused to the transmembrane domain of syntaxin, were greatly reduced or not detectable, respectively. In acutely light-adapted rods, an arrestin-eGFP fusion protein also exhibited axial variation. Both the light-sensitive Rho-eGFP and arrestin-eGFP banding were in phase with the previously characterized birefringence banding (Kaplan, Invest. Ophthalmol. Vis. Sci. 21, 395-402 1981). In contrast, endogenous rhodopsin did not exhibit such axial variation. Thus, there is an axial inhomogeneity in membrane composition or structure, detectable by the rhodopsin transgene density distribution and regulated by the light cycle, implying a light-regulated step for disk assembly in the OS. The impact of these results on the use of chimeric proteins with rhodopsin fused to fluorescent proteins at the carboxyl terminus is discussed.


A randomized trial of daily awakening in critically ill patients managed with a sedation protocol: a pilot trial.
Protocolized sedation (PS) and daily sedative interruption (DI) in critically ill patients have both been shown to shorten the durations of mechanical ventilation (MV) and intensive care unit (ICU) stay. Our objective was to determine the safety and feasibility of a randomized trial to determine whether adults managed with both PS + DI have a shorter duration of MV than patients managed with PS alone. Prospective randomized, concealed, unblinded, multicenter, pilot trial. Three university-affiliated medical-surgical ICUs. Sixty-five adults anticipated to require MV >48 hrs and receiving sedative/analgesic infusions. Patients were randomized to PS alone, or PS + DI. PS was implemented by bedside nurses; sedatives/analgesics were titrated to achieve Sedation Agitation Score (SAS) 3-4. The PS + DI group also had infusions interrupted daily until the patients awoke. Diagnosis, age [mean +/- SD] (53 +/- 18.3 vs. 62.1 +/- 16.7 yrs) and Acute Physiology and Chronic Health Evaluation II (27.7 +/- 8.4 vs. 26.6 +/- 8.4) were similar in the PS and PS + DI groups, respectively. The median duration of MV in the PS and PS + DI groups was 8.0 vs. 10.5 days, and ICU stay was 10.0 vs. 13.0 days, respectively. The SAS was within target range (3-4) in 59% of 9,611 measurements, and within an acceptable range (2-5) in 86% of measurements. Self-assessed nursing and respiratory therapist workload was low in the majority of the cohort. Adverse events were similar in both groups. Patient recruitment was slower than projected (1.5 patients/mo). This pilot trial comparing PS vs. PS + DI confirmed the safety and acceptability of the sedation protocol and DI, and guided important modifications to the protocol, thus enhancing the feasibility of a future multicenter trial. This trial was not designed to detect small but significant differences in clinically important outcomes.


Extracellular Vesicles for Research on Psychiatric Disorders.
Extracellular vesicles (EVs) have gained increasing attention as underexplored intercellular communication mechanisms in basic science and as potential diagnostic tools in translational studies, particularly those related to cancers and neurological disorders. This article summarizes accumulated findings in the basic biology of EVs, EV research methodology, and the roles of EVs in brain cell function and dysfunction, as well as emerging EV studies in human brain disorders. Further research on EVs in neurobiology and psychiatry may open the door to a better understanding of intercellular communications in healthy and diseased brains, and the discovery of novel biomarkers and new therapeutic strategies in psychiatric disorders.


Small-for-Size Syndrome: Bridging the Gap Between Liver Transplantation and Graft Recovery.
In living donor liver transplantation, optimal graft size is estimated from values like graft volume/standard liver volume and graft/recipient body weight ratio but the final functional hepatic mass is influenced by other donor and recipient factors. Grafts with insufficient functional hepatic mass can produce a life-threatening condition with rapidly progressive liver failure called small-for-size syndrome (SFSS). Diagnosis of SFSS requires careful surveillance for signs of inadequate hepatocellular function, residual portal hypertension, and systemic inflammation that suggest rapidly progressive liver failure. Early diagnosis, symptom control, and addressing the cause of SFSS may prevent the need for retransplantation. With increased attention to avoiding donor risk, intensivists will be confronted with more SFSS recipients. In this review, we aim to outline a systematic approach to the medical management of patients with SFSS by providing a concise synopsis of general supportive care-neurological, cardiovascular, and renal support, mechanical ventilation, nutritional support, infection control, and tailored immunosuppression-with an aim to avoid end-organ damage or death and a review of current interventions including liver support devices, portal flow modulating drugs, and other experimental interventions that aim to preserve existing hepatic mass and improve conditions for hepatic regeneration. We examine evidence for SFSS interventions to provide the reader with information that may assist in clinical decision making. Points of controversy in care are purposefully highlighted to identify areas where additional experimental work is still needed. A full understanding of the pathophysiology of SFSS and measures to support liver regeneration will guide effective management.


Dietary indicators of oral and pharyngeal cancer.
The relationship between frequency of consumption of a selected number of indicator foods and oral and oropharyngeal cancer risk was analysed in a case-control study conducted in Northern Italy on 105 cases of oral and pharyngeal cancer and 1169 controls in hospital for acute, non-neoplastic or digestive diseases. Besides significant and strong direct associations with tobacco (relative risk, RR = 11.0 for current versus never smokers) and alcohol (RR = 5.8 for upper versus lower consumption tertile), consumption of six food items (milk, meat, cheese, carrots, green vegetables and fruit) were inversely and significantly related to oral and pharyngeal cancer risk. The strongest protection was apparently related to frequent fruit consumption, with RRs of 0.8 and 0.2 in the two highest tertiles. Allowance for major potential confounding factors, including tobacco, alcohol and social class indicators explained only part of the dietary correlates observed. The two items remaining significant after multivariate analysis were fruit (RR = 0.3 for the upper tertile) and alcohol (RR = 3.8 for the upper tertile). The associations observed may simply reflect a generally poorer nutritional status in the cases, although the observation that fruit consumption appears to be a particularly important protective factor against oropharyngeal cancer is of potential interest, in terms of aetiological clues and preventive implications.


Blood miR-21 and miR-26 tailor a good diagnostic model for childhood asthma.
This study was to investigate the relationships between blood miR-21/26a with the prevalence and severity of childhood asthma (CAMP). For this purpose, 123 children with allergic asthma (AZ) from June 2018 to June 2020, and 60 contemporaneous healthy children for reference, were enrolled. Lung function was detected using a portable pediatric spirometer and AZ severity was evaluated. Blood samples of admissions were collected to quantify the expression degrees of miR-21 and miR-26a. Logistic regression analysis and model were constructed. Results showed that (1) CAMP had higher MiR-21 expression and lower MiR-26a expression than healthy controls; (2) The severity of AZ, evidenced by FEV1/PV, significantly correlated with miR-21(Y=-3.825X+102.6, P<0.001) and miR-26a (Y=10.43X+54.29, P<0.001); (3) The prevalence of AZ-related to miR-21 (OR=4.180, P<0.001) and miR-26a (OR=0.058, P<0.001) after adjusting for cofounders. (4) the expression levels of miR-21/26a had a good diagnostic potential for AZ (AUC are 0.85 and 0.94, respectively). In conclusion, Blood miRNA-21 and miR-26a are promising biomarkers for the diagnosis and severity of CAMP.


Genomic alterations in cultured human embryonic stem cells.
Cultured human embryonic stem cell (hESC) lines are an invaluable resource because they provide a uniform and stable genetic system for functional analyses and therapeutic applications. Nevertheless, these dividing cells, like other cells, probably undergo spontaneous mutation at a rate of 10(-9) per nucleotide. Because each mutant has only a few progeny, the overall biological properties of the cell culture are not altered unless a mutation provides a survival or growth advantage. Clonal evolution that leads to emergence of a dominant mutant genotype may potentially affect cellular phenotype as well. We assessed the genomic fidelity of paired early- and late-passage hESC lines in the course of tissue culture. Relative to early-passage lines, eight of nine late-passage hESC lines had one or more genomic alterations commonly observed in human cancers, including aberrations in copy number (45%), mitochondrial DNA sequence (22%) and gene promoter methylation (90%), although the latter was essentially restricted to 2 of 14 promoters examined. The observation that hESC lines maintained in vitro develop genetic and epigenetic alterations implies that periodic monitoring of these lines will be required before they are used in in vivo applications and that some late-passage hESC lines may be unusable for therapeutic purposes.