Immunocytochemical studies on the localization of plasma and of cellular retinol-binding proteins and of transthyretin (prealbumin) in rat liver and kidney.
The immunocytochemical localization of cellular retinol-binding protein (CRBP), of plasma retinol-binding protein (RBP), and of plasma transthyretin (TTR) was studied in rat liver and kidney. The studies employed normal rats, retinol-deficient rats, and rats fed excess retinol. Antisera were prepared in rabbits against purified rat CRBP, RBP, and TTR. The primary antibodies and goat anti-rabbit IgG were purified by immunosorbent affinity chromatography, using the respective pure antigen coupled to Sepharose as the immunosorbent. This procedure effectively removed cross-reactive and heterophile antibodies, which permitted the specific staining and localization of each antigen by the unlabeled peroxidase-antiperoxidase method. CRBP was found to be localized in two cell types in the liver, the parenchymal cells and the fat-storing cells. Diffuse cytoplasmic staining for CRBP was seen in all the parenchymal cells. Much more intense staining for CRBP was seen in the fat-storing cells. The prominence of the CRBP-positive fat-storing cells changed markedly with vitamin A status. Thus, these cells were most prominent, and appeared most numerous, in liver from rats fed excess retinol. Both RBP and TTR were localized within liver parenchymal cells. The intensity of RBP staining increased markedly in retinol-deficient rat liver, consistent with previous biochemical observations. With the methods employed, specific staining for RBP or TTR was not seen in cells other than the parenchymal cells. In the kidney, all three proteins (CRBP, RBP, and TTR) were localized in the proximal convoluted tubules of the renal cortex. Staining for RBP was much more intense in normal kidney than in kidney from retinol-deficient rats. These findings reflect the fact that RBP in the tubules represents filtered and reabsorbed RBP. The pattern of specific staining for CRBP among the various tubules was very similar to that seen for RBP on adjacent, serial sections of kidney. The function of CRBP in the kidney is not known.


Comparison of Nonabsorbable and Absorbable Suture in Total Knee Arthroplasty.
BACKGROUND Wound closure of KA is important for postoperative rehabilitation. At present there is still no consensus on the best wound closure technique for KA. We performed the present study to determine whether absorbable suture is better than nonabsorbable suture in total knee arthroplasty (TKA). MATERIAL AND METHODS A total of 180 patients who underwent TKA were divided into 3 groups: 80 cases of nonabsorbable suture, 50 cases of 2-0 absorbable suture, and 50 cases of 4-0 absorbable suture. The time required for closure, frequency of gauze change, length of stay in hospital, adverse events, range of motion (ROM) after 3 months postoperatively, and VAS score of wounds were calculated. Comparison was made to explore any significant differences between different groups. RESULTS There were significant differences between the nonabsorbable group and the absorbable group with regards to closure time, frequency of gauze change, and hospital length of stay (LOS). Closure time was longer in the absorbable group than in the nonabsorbable group. Frequency of gauze change, hospital LOS, and adverse events were lower, and VAS was higher in the absorbable group. Closure time was longer in the 4-0 absorbable group than in the 2-0 group. There was no significant difference between the 4-0 group and 2-0 group in other variables. There was no significant difference in long-term ROM among all groups. CONCLUSIONS Absorbable suture in TKA reduces the incidence of fatty liquefaction, frequency of gauze change, and postoperative LOS. It improves the cosmetic appearance and overall reduces the economic cost. There was no significant effect on early and long-term functional ROM. In conclusion, absorbable suture can be used in TKA when appropriately indicated.


New genetic and morphological evidence suggests a single hoaxer created 'Piltdown man'.
In 1912, palaeontologist Arthur Smith Woodward and amateur antiquarian and solicitor Charles Dawson announced the discovery of a fossil that supposedly provided a link between apes and humans: Eoanthropus dawsoni (Dawson's dawn man). The publication generated huge interest from scientists and the general public. However, 'Piltdown man's' initial celebrity has long been overshadowed by its subsequent infamy as one of the most famous scientific frauds in history. Our re-evaluation of the Piltdown fossils using the latest scientific methods (DNA analyses, high-precision measurements, spectroscopy and virtual anthropology) shows that it is highly likely that a single orang-utan specimen and at least two human specimens were used to create the fake fossils. The modus operandi was found consistent throughout the assemblage (specimens are stained brown, loaded with gravel fragments and restored using filling materials), linking all specimens from the Piltdown I and Piltdown II sites to a single forger-Charles Dawson. Whether Dawson acted alone is uncertain, but his hunger for acclaim may have driven him to risk his reputation and misdirect the course of anthropology for decades. The Piltdown hoax stands as a cautionary tale to scientists not to be led by preconceived ideas, but to use scientific integrity and rigour in the face of novel discoveries.


Drug misuse in a special clinic patient population in Glasgow.
During the period October, 1970, to October, 1972, a sample of 295 patients attending Special Clinics in the City of Glasgow participated in an investigation into drug misuse. Results show that, of the 136 males and 159 females who took part--all within an age range of 16 to 24 years, 43 per cent. of the males and 36 per cent. of the females admitted to having experimented with drugs. These percentage figures on the prevalence of drug misuse compare with 36 per cent. of males and 24 per cent. of females obtained from other Glasgow-domiciled target groups, comprising a sample of 2,514 individuals. The principal drug of misuse is shown to be cannabis, followed by LSD and sleeping pills. Data on drug availability suggest that little difficulty is experienced in obtaining drugs illicitly from a variety of sources. Comparison between drug misusers and non-misusers on domestic, educational, and social variables revealed no very distinctive patterns. Results of screening for personality characteristics, using Eysenck's Psychoticism, Extraversion and Neuroticism Inventory generally confirmed previous findings related to atypical scores on the P, E, and N parameters. Even so, results obtained by means of the PEN Inventory and IPAT Anxiety Scale revealed a pattern in which drug misuse is shown to be related to even higher scores of psychoticism, neuroticism, and anxiety. The results of the investigation are discussed with particular reference to the nature of the appeal of drugs to the sexually permissive and promiscuous.


Immediate heart-rate response to standing: simple test for autonomic neuropathy in diabetes.
The immediate heart-rate response to standing was measured in 22 normal controls and 25 patients with diabetes, 15 of whom had autonomic neuropathy. The response in the controls and patients without autonomic neuropathy was characteristic and consistent, with tachycardia maximal at around the 15th beat and relative bradycardia maximal at around the 30th beat. The diabetics with autonomic neuropathy, however, showed a flat response. In three controls the response was abolished with intravenous atropine but not with propranolol, showing that it is mediated through the vagus. A simplified test using routine ECGs and measuring the R-R interval at beats 15 and 30 with a ruler is easily performed as an outpatient procedure and may be used as a measure of autonomic function in diabetes.


Meta-analysis of new genome-wide association studies of colorectal cancer risk.
Colorectal cancer is the second leading cause of cancer death in developed countries. Genome-wide association studies (GWAS) have successfully identified novel susceptibility loci for colorectal cancer. To follow up on these findings, and try to identify novel colorectal cancer susceptibility loci, we present results for GWAS of colorectal cancer (2,906 cases, 3,416 controls) that have not previously published main associations. Specifically, we calculated odds ratios and 95% confidence intervals using log-additive models for each study. In order to improve our power to detect novel colorectal cancer susceptibility loci, we performed a meta-analysis combining the results across studies. We selected the most statistically significant single nucleotide polymorphisms (SNPs) for replication using ten independent studies (8,161 cases and 9,101 controls). We again used a meta-analysis to summarize results for the replication studies alone, and for a combined analysis of GWAS and replication studies. We measured ten SNPs previously identified in colorectal cancer susceptibility loci and found eight to be associated with colorectal cancer (p value range 0.02 to 1.8 × 10(-8)). When we excluded studies that have previously published on these SNPs, five SNPs remained significant at p < 0.05 in the combined analysis. No novel susceptibility loci were significant in the replication study after adjustment for multiple testing, and none reached genome-wide significance from a combined analysis of GWAS and replication. We observed marginally significant evidence for a second independent SNP in the BMP2 region at chromosomal location 20p12 (rs4813802; replication p value 0.03; combined p value 7.3 × 10(-5)). In a region on 5p33.15, which includes the coding regions of the TERT-CLPTM1L genes and has been identified in GWAS to be associated with susceptibility to at least seven other cancers, we observed a marginally significant association with rs2853668 (replication p value 0.03; combined p value 1.9 × 10(-4)). Our study suggests a complex nature of the contribution of common genetic variants to risk for colorectal cancer.


Aberrant Activity in Degenerated Retinas Revealed by Electrical Imaging.
In this review, I present and discuss the current understanding of aberrant electrical activity found in the ganglion cell layer (GCL) of rod-degenerated (rd) mouse retinas. The reported electrophysiological properties revealed by electrical imaging using high-density microelectrode arrays can be subdivided between spiking activity originating from retinal ganglion cells (RGCs) and local field potentials (LFPs) reflecting strong trans-membrane currents within the GCL. RGCs in rd retinas show increased and rhythmic spiking compared to age-matched wild-type retinas. Fundamental spiking frequencies range from 5 to 15 Hz in various mouse models. The rhythmic RGC spiking is driven by a presynaptic network comprising AII amacrine and bipolar cells. In the healthy retina this rhythm-generating circuit is inhibited by photoreceptor input. A unique physiological feature of rd retinas is rhythmic LFP manifested as spatially-restricted low-frequency (5-15 Hz) voltage changes. Their spatiotemporal characterization revealed propagation and correlation with RGC spiking. LFPs rely on gap-junctional coupling and are shaped by glycinergic and by GABAergic transmission. The aberrant RGC spiking and LFPs provide a simple readout of the functionality of the remaining retinal circuitry which can be used in the development of improved vision restoration strategies.


Distribution patterns of mucosally applied particles and characterization of the antigen presenting cells.
Mucosal application is the most common route of vaccination to prevent outbreaks of infectious diseases like Newcastle disease virus (NDV). To gain more knowledge about distribution and uptake of a vaccine after mucosal vaccination, we studied the distribution pattern of antigens after different mucosal routes of administration. Chickens were intranasally (i.n.), intratracheally (i.t.) or intraocularly (i.o.) inoculated with fluorescent beads and presence of beads in nasal-associated lymphoid tissue (NALT), Harderian gland (HG), conjunctiva-associated lymphoid tissue (CALT), trachea, lungs, air sacs, oesophagus and blood was characterized. The distribution patterns differed significantly between the three inoculation routes. After i.t. inoculation, the beads were mainly retrieved from trachea, NALT and lung. I.n. inoculation resulted in beads found mainly in NALT but detectable in all organs sampled. Finally, after i.o. inoculation, the beads were detected in NALT, CALT, HG and trachea. The highest number of beads was retrieved after i.n. inoculation. Development of novel vaccines requires a comprehensive knowledge of the mucosal immune system in birds in order to target vaccines appropriately and to provide efficient adjuvants. The NALT is likely important for the induction of mucosal immune responses. We therefore studied the phenotype of antigen-presenting cells isolated from NALT after i.n. inoculation with uncoated beads or with NDV-coated beads. Both types of beads were efficiently taken up and low numbers of bead+ cells were detected in all organs sampled. Inoculation with NDV-coated beads resulted in a preferential uptake by NALT antigen-presenting cells as indicated by high percentages of KUL01+-, MHC II+ and CD40+ bead+ cells.


Advertising Content, Platform Characteristics and the Appeal of Beer Advertising on a Social Networking Site.
The current study was conducted to investigate how changes in the content of a social media ad, user engagement values associated with the ad and user-generated comments (UGCs) associated with the ad can influence the appeal (i.e. source appeal, informational appeal and emotional appeal) of a social media ad. Facebook beer ads that violated the guidelines of a relevant marketing code were rated as more emotionally appealing compared to Facebook beer ads that did not violated the guidelines. Increased emotional appeal in beer advertising increases the probability that the ad will be remembered and influence future drinking occasions. A 2 (ad regulatory compliance: compliant vs. non-compliant) × 2 (user engagement: low vs. high) × 2 (UGC congruence: pro- vs anti-alcohol) mixed factorial experiment was conducted with 120 young adults, 21-24 years old. Each participant viewed four Facebook beer ads that were previously evaluated for thematic content and regulatory compliance. Participants were randomized to view either high or low user engagement values and either pro- or anti-drinking user-generated comments. After each ad exposure, ad appeal was assessed. Statistical analysis was conducted using hierarchical linear modeling. Models were adjusted for demographics, Alcohol Use Disorders Identification Test (AUDIT) scores and Facebook involvement. Source appeal (P = 0.034) and informational appeal (P < 0.001) were significantly higher among ads that were compliant with existing advertising regulations. Emotional appeal was significantly higher among ads that were non-compliant (P = 0.004). The effect of user engagement and UGCs were non-significant (p's > 0.05). Additionally, AUDIT scores (p's < 0.01) and Facebook involvement scores (p's < 0.01) were positively associated with all forms of ad appeal. The appeal of Facebook beer ads may be primarily determined by ad content. Increased emotional appeal in advertising caused by non-compliant advertising may increase the probability that the ad will be remembered and influence future drinking occasions.


SirT1 gain of function increases energy efficiency and prevents diabetes in mice.
In yeast, worms, and flies, an extra copy of the gene encoding the Sirtuin Sir2 increases metabolic efficiency, as does administration of polyphenols like resveratrol, thought to act through Sirtuins. But evidence that Sirtuin gain of function results in increased metabolic efficiency in mammals is limited. We generated transgenic mice with moderate overexpression of SirT1, designed to mimic the Sirtuin gain of function that improves metabolism in C. elegans. These mice exhibit normal insulin sensitivity but decreased food intake and locomotor activity, resulting in decreased energy expenditure. However, in various models of insulin resistance and diabetes, SirT1 transgenics display improved glucose tolerance due to decreased hepatic glucose production and increased adiponectin levels, without changes in body weight or composition. We conclude that SirT1 gain of function primes the organism for metabolic adaptation to insulin resistance, increasing hepatic insulin sensitivity and decreasing whole-body energy requirements. These findings have important implications for Sirtuin-based therapies in humans.


Classification of crystallization outcomes using deep convolutional neural networks.
The Machine Recognition of Crystallization Outcomes (MARCO) initiative has assembled roughly half a million annotated images of macromolecular crystallization experiments from various sources and setups. Here, state-of-the-art machine learning algorithms are trained and tested on different parts of this data set. We find that more than 94% of the test images can be correctly labeled, irrespective of their experimental origin. Because crystal recognition is key to high-density screening and the systematic analysis of crystallization experiments, this approach opens the door to both industrial and fundamental research applications.


Four Decades of Obesity Trends among Non-Hispanic Whites and Blacks in the United States: Analyzing the Influences of Educational Inequalities in Obesity and Population Improvements in Education.
Both obesity (body mass index ≥ 30) and educational attainment have increased dramatically in the United States since the 1970s. This study analyzed the influences of educational inequalities in obesity and population improvements in education on national obesity trends between 1970 and 2010. For non-Hispanic white and black males and females aged 25-74 years, educational differences in the probability of being obese were estimated from the 1971-2012 National Health and Nutrition Examination Surveys, and population distributions of age and educational groups, from the 1970 Census and 2010 American Community Survey. In the total population, obesity increased from 15.7% to 38.8%, and there were increases in the greater obese probabilities of non-college graduates relative to four-year college graduates. The increase in obesity would have been lower by 10% (2.2 percentage points) if educational inequalities in obesity had stayed at their 1970 values and lower by one third (7.9 points) if obesity inequalities had been eliminated. Obesity inequalities were larger for females than males and for whites than blacks, and obesity did not differ by education among black males. As a result, the impact of obesity inequalities on the obesity trend was largest among white females (a 47% reduction in the obesity increase if obesity inequalities had been eliminated), and virtually zero among black males. On the other hand, without educational improvements, the obesity increase would have been 9% more in the total population, 23% more among white females and not different in the other three subpopulations. Results indicate that obesity inequalities made sizable contributions to the obesity trends, and the obesity reductions associated with educational improvements were more limited.


Adult Anomalous Left Coronary Artery Arising From the Pulmonary Artery (ALCAPA) Syndrome as First Presentation With Atrial Fibrillation in a Marathon Runner.
Anomalous left coronary artery arising from the pulmonary artery (ALCAPA) syndrome is a very rare congenital heart disease with an incidence of one in 300,000 and a high rate of mortality early in life if left untreated. Adult-type ALCAPA presents when significant collaterals develop from the right coronary artery (RCA) to the left coronary artery (LCA). Even with the collaterals, chronic sub-endocardial ischemia occurs in most cases, and patients die from sudden cardiac death. Here we present a case of a 38-year-old female who lived an active and healthy life and presented with chest pain and palpitations. Initial electrocardiography (EKG) showed atrial fibrillation with rapid ventricular response. Although initial cardiac enzymes were negative and there were no ischemic EKG changes, troponins became elevated over the course of the hospital stay and the patient underwent a left heart catheterization. Results revealed a dilated RCA extending to the left ventricle and an anomalous left main originating from the pulmonary artery with complete occlusion. The patient received medical management for acute coronary syndrome, including heparin infusion for 48 hours, aspirin, metoprolol, and atorvastatin. She was referred to a tertiary care facility for surgical correction of anomaly of the coronary arteries. The next day, the patient arrived in the emergency department with an acute onset of speech difficulty and left-sided weakness. A brain CT without contrast showed hematoma in the right frontal lobe. The patient underwent surgical evacuation of the hematoma with marked improvement of her weakness. The patient recovered after a successful surgical repair involving translocation of the left main coronary artery to the aorta. It has been reported that ALCAPA should be considered in a young adult with dilated cardiomyopathy and mitral regurgitation (MR). Other common presentations include acute myocardial infarction, angina, and dyspnea on exertion. Sudden cardiac death is not uncommon; however, it tends to decrease with age of diagnosis. Interestingly, our patient was known to have MR with regular follow-up at the cardiology clinic for years. Echocardiogram never showed any abnormalities other than MR. She never received further workup to address the reason of MR, although she has no underlying chronic conditions that can explain it. In relatively young patients with a healthy lifestyle presenting with chest pain, a broader look at etiologies should be considered. We would like to emphasize the importance of looking up for possible coronary artery disease, especially in young individuals.


Cue-dependent effects of VR experience on motion-in-depth sensitivity.
The visual system exploits multiple signals, including monocular and binocular cues, to determine the motion of objects through depth. In the laboratory, sensitivity to different three-dimensional (3D) motion cues varies across observers and is often weak for binocular cues. However, laboratory assessments may reflect factors beyond inherent perceptual sensitivity. For example, the appearance of weak binocular sensitivity may relate to extensive prior experience with two-dimensional (2D) displays in which binocular cues are not informative. Here we evaluated the impact of experience on motion-in-depth (MID) sensitivity in a virtual reality (VR) environment. We tested a large cohort of observers who reported having no prior VR experience and found that binocular cue sensitivity was substantially weaker than monocular cue sensitivity. As expected, sensitivity was greater when monocular and binocular cues were presented together than in isolation. Surprisingly, the addition of motion parallax signals appeared to cause observers to rely almost exclusively on monocular cues. As observers gained experience in the VR task, sensitivity to monocular and binocular cues increased. Notably, most observers were unable to distinguish the direction of MID based on binocular cues above chance level when tested early in the experiment, whereas most showed statistically significant sensitivity to binocular cues when tested late in the experiment. This result suggests that observers may discount binocular cues when they are first encountered in a VR environment. Laboratory assessments may thus underestimate the sensitivity of inexperienced observers to MID, especially for binocular cues.


Ethnicity and ERG frequency in prostate cancer.
Emerging observations emphasize a distinct biology of prostate cancer among men of different ethnicities and races, as demonstrated by remarkable differences in the frequency of ERG oncogenic activation, one of the most common and widely studied prostate cancer driver genes. Worldwide assessment of ERG alterations frequencies show consistent trends, with men of European ancestry having the highest rates of alteration and men of African or Asian ancestries having considerably lower alteration rates. However, data must be interpreted cautiously, owing to variations in assay platforms and specimen types, as well as ethnic and geographical classifications. Many opportunities and challenges remain in assessing cancer-associated molecular alterations at a global level, and these need to be addressed in order to realize the true potential of precision medicine for all cancer patients.


Yerba Maté (Illex Paraguariensis) ingestion augments fat oxidation and energy expenditure during exercise at various submaximal intensities.
Ingesting Yerba Maté (YM) has become widely popular for health promotion, obesity prevention and body weight reduction, primarily due its thermogenic effectiveness. However, the YM effects on fat metabolism during exercise, when fat metabolism is already increased several fold, are unknown. The present study investigated whether acute YM ingestion augments fat metabolism parameters of fatty acid oxidation (FAO) and energy expenditure derived from FAO (EEFAO) during exercise with several intensities. Fourteen healthy males and females were randomised in a repeated measures crossover experimental design. All participants ingested either 1000 mg of YM or placebo capsules (PLC) 60 min before performing two incremental exercise ergometry tests. Power output was initiated at and increased by 0.5 W.kg(-1) of body weight every 3 min stage, until reaching peak oxygen uptake [Formula: see text]. Expired gases and stoichiometric indirect calorimetry were used to analyse FAO and EEFAO. Capillary blood samples were collected and analysed for blood lactate concentration (BLC) at rest and at each submaximal and maximal power output. YM significantly increased FAO and EEFAO by 24% in all submaximal exercise intensities below 70% of [Formula: see text] (p < 0.001, ANOVA main effects) with post hoc tests showing a higher FAO and EEFAO (p < 0.05) at the lower exercise intensities (e.g. 0.26 ± 0.09 vs. 0.35 ± 0.10 and 0.25 ± 0.12 vs. 0.33 ± 0.11 g.min(-1) at 40 and 50% of [Formula: see text] respectively). These changes were combined with a trend towards a decrease in BLC (P = 0.066), and without a significant difference in [Formula: see text], peak power, peak RER, or peak BLC. Acute YM ingestion augments the exercise dependent increase in FAO and EEFAO at submaximal exercise intensities without negatively affecting maximal exercise performance, suggesting a potential role for YM ingestion to increase the exercise effectiveness for weight loss and sports performance.


Survey of alternative medicine use among organ transplant patients.
Herbal medicine and health food supplements have become increasingly popular. However, many of these pharmacologically active compounds remain poorly understood. Patients with chronic and life-threatening conditions often use alternative therapies while receiving conventional medical care, and his population is at increased risk for complications and adverse drug interactions due to poor health and complex drug regimens. Patients awaiting or who had received solid organ transplants were surveyed about their use of herbal medicines and health food supplements. Twenty percent of respondents acknowledged experience with these products, which they used to prolong the function of a failing organ or to obtain relief from fatigue and insomnia. Transplant staff often were unaware of their patients' use of these treatments, despite patients' claims to the contrary. The potential for unexpected drug interactions, toxicity, and other adverse reactions resulting from the use of herbal medicines or supplements must be recognized and identified by transplant teams.


Preliminary results of autologous adipose-derived stem cells in early knee osteoarthritis: identification of a subpopulation with greater response.
The purpose of this study was to report the clinical and functional results of a series of patients with early knee osteoarthritis (KOA) treated with the intra-articular injection of autologous adipose-derived stem cells (aASCs) plus arthroscopic debridement. The hypothesis was that protocol would significantly improve the clinical and functional outcomes in patients with early KOA. Fifty-two patients with early KOA, who received arthroscopic debridement followed by percutaneous injection of aASCs, were enrolled into the study and retrospectively analyzed with an average follow-up of 15.3 (range, 6 to 24) months. Patients were assessed through the IKS knee and function scores and VAS pain scale. The mean IKS knee score improved from 37.4 (range, 14 to 79) points pre-operatively to 62.6 (range, 27 to 95) points at the latest follow-up (p < < 0.01). The mean IKS function score improved from 57.2 (range, 25 to 100) points pre-operatively to 83.0 (range, 35 to 100) points at the latest follow-up (p < < 0.01). The mean VAS score decreased from 8.5 (range, 3 to 10) pre-operatively to 5.1 (range, 0 to 8) at the latest follow-up (p < < 0.01). Additionally, patients with a pre-operative VAS score greater than 8 were found to show greater clinical and functional benefits compared with patients with VAS score lower than 8. The knee injection of aASCs associated to arthroscopic debridement increased significantly the clinical and functional scores in patients with early KOA at a mid-term follow-up, especially those with higher pre-operative VAS scores.


Seeing responsibility: can neuroimaging teach us anything about moral and legal responsibility?
As imaging technologies help us understand the structure and function of the brain, providing insight into human capabilities as basic as vision and as complex as memory, and human conditions as impairing as depression and as fraught as psychopathy, some have asked whether they can also help us understand human agency. Specifically, could neuroimaging lead us to reassess the socially significant practice of assigning and taking responsibility? While responsibility itself is not a psychological process open to investigation through neuroimaging, decision-making is. Over the past decade, different researchers and scholars have sought to use neuroimaging (or the results of neuroimaging studies) to investigate what is going on in the brain when we make decisions. The results of this research raise the question whether neuroscience-especially now that it includes neuroimaging-can and should alter our understandings of responsibility and our related practice of holding people responsible. It is this question that we investigate here.


Isolation of immunomodulatory triterpene acids from a standardized rose hip powder (Rosa canina L.).
A previously published systematic review and a metaanalysis have concluded that the consumption of standardized rose hip powder (Rosa canina L.) can reduce pain in osteoarthritis patients. Synovial inflammation has been suggested to play an important role in the pathogenesis of osteoarthritis and mainly to involve infiltration of the synovial membrane by macrophages. Therefore, the immunomodulatory effect of standardized rose hip powder of Rosa canina L. was investigated and active principles isolated using the Mono Mac 6 cell line as a model for human macrophages. Treatment of Mono Mac 6 cells with the residue of a crude dichloromethane extract of rose hip powder significantly and concentration dependently inhibited the lipopolysaccharide induced interleukin-6 release. Through bioassay-guided fractionation the immunomodulatory effect of the dichloromethane extract was correlated to a mixture of three triterpene acids; oleanolic acid, betulinic acid and ursolic acid (IC(50) 21 ± 6 µm). Further studies revealed that only oleanolic acid and ursolic acid, but not betulinic acid, could inhibit the lipopolysaccharide induced interleukin-6 release from Mono Mac 6 cells when tested separately. Combination of either oleanolic acid or ursolic acid with betulinic acid enhanced the immunomodulatory effect of the two triterpene acids.


Use of targeted glycoproteomics to identify serum glycoproteins that correlate with liver cancer in woodchucks and humans.
Chronic infection with hepatitis B virus (HBV) is associated with the majority of hepatocellular carcinoma (HCC). The diagnosis of HCC is usually made in the late stages of the disease, when treatment options are limited and prognosis is poor. We therefore have developed a method of glycoproteomic analysis in an attempt to discover serum markers that can assist in the early detection of HBV-induced liver cancer. Briefly, a comparative method for analysis of oligosaccharides released from serum glycoproteins and for recovery and identification of proteins with aberrant glycosylation, as a function of cancer diagnosis, is described. The model we have used is the woodchuck (Marmota monax), which shares similarities in the glycosylation pattern associated with liver proteins in human HCC. In this report, we show that woodchucks diagnosed with HCC have dramatically higher levels of serum-associated core alpha-1,6-linked fucose, as compared with woodchucks without a diagnosis of HCC. The coupling of this methodology with 2D gel proteomics has permitted the identification of several glycoproteins with altered glycosylation as a function of cancer. One such glycoprotein, Golgi Protein 73 (GP73), was found to be elevated and hyperfucosylated in animals with HCC. Further, the study showed GP73 to be elevated in the serum of people with a diagnosis of HCC, providing a validation of our approach. The potential of this technology for biomarker discovery and the implications of increased levels of GP73 in liver cancer are discussed.


Does an occluded RCA affect prognosis in patients undergoing PCI or CABG for left main coronary artery disease? Analysis from the EXCEL trial.
The impact of an occluded right coronary artery (RCA) in patients with left main coronary artery disease (LMCAD) undergoing revascularisation is unknown. We compared outcomes for patients with LMCAD randomised to percutaneous coronary intervention (PCI) or coronary artery bypass grafting (CABG) according to the presence of an occluded RCA in the EXCEL trial. The EXCEL trial randomised 1,905 patients with LMCAD and SYNTAX scores ≤32 to PCI with everolimus-eluting stents versus CABG. Patients were categorised according to whether they had an occluded RCA at baseline, and their outcomes were examined using multivariable Cox proportional hazards regression. The primary endpoint was a composite of death, stroke, or myocardial infarction at three years. Among 1,753 patients with a dominant RCA by core laboratory analysis, the RCA was occluded in 130 (7.4%) at baseline. PCI was attempted in 34 of 65 patients with an occluded RCA (52.3%) and was successful in 27 (79.4% of those attempted; 41.5% of all RCAs recanalised). The RCA was bypassed in 42 of 65 patients with an occluded RCA (64.6%; p=0.0008 versus PCI). The three-year absolute and relative rates of the primary endpoint were similar between PCI and CABG, in patients with or without an occluded RCA (pinteraction=0.92). In the EXCEL trial, the presence of an occluded RCA at baseline did not confer a worse three-year prognosis in patients undergoing revascularisation for LMCAD and did not affect the relative outcomes of PCI versus CABG in this high-risk patient cohort.


Family history in participants of breast cancer screening.
Of 12,337 participants of mass screening for breast cancer (screenees) in Gunma Prefecture between 1980 to 1985, 1,000 participants' records in Gunma Prefecture were investigated. The records of screenees who had already developed breast cancer were excluded. The rate of screenees who had a family history (FH) of cancer in first- and second-degree relatives was compared with that of 1,248 breast cancer patients (controls) using the conditional logistic regression model. The mean age of screenees (49 y.o.) was similar to that of controls (51 y.o.). The number of screenees with positive FH of cancer including other malignancies was 530 with the odds ratio of 2.68. This was statistically significant with chi-square test. The rates of screenees with positive FH of cancer were quite similar in the seven districts. Screenees had a significantly larger number of relatives with a positive FH of breast cancer compared with the controls. The rate of positive breast cancer history in parents of screenees was significantly higher than in the parents of controls. Rates of positive breast cancer history in grandmothers and aunts of screenees were also significantly higher than that in the controls. The rate of breast cancer history in sisters was not different between the two groups. Screenees had a higher rate of positive FH of cancer in parents, especially in mothers. Family histories of stomach, liver, lung, and uterus cancer in parents were more frequently observed in screenees compared with the controls and those cancer histories were also frequently observed in other family members of screenees. These data showed that a FH of cancer is one of the primary motivations of participation in the breast cancer screening program. Participants seemed to be intrinsically a high risk group of breast cancer.


Deep sparse multi-task learning for feature selection in Alzheimer's disease diagnosis.
Recently, neuroimaging-based Alzheimer's disease (AD) or mild cognitive impairment (MCI) diagnosis has attracted researchers in the field, due to the increasing prevalence of the diseases. Unfortunately, the unfavorable high-dimensional nature of neuroimaging data, but a limited small number of samples available, makes it challenging to build a robust computer-aided diagnosis system. Machine learning techniques have been considered as a useful tool in this respect and, among various methods, sparse regression has shown its validity in the literature. However, to our best knowledge, the existing sparse regression methods mostly try to select features based on the optimal regression coefficients in one step. We argue that since the training feature vectors are composed of both informative and uninformative or less informative features, the resulting optimal regression coefficients are inevidently affected by the uninformative or less informative features. To this end, we first propose a novel deep architecture to recursively discard uninformative features by performing sparse multi-task learning in a hierarchical fashion. We further hypothesize that the optimal regression coefficients reflect the relative importance of features in representing the target response variables. In this regard, we use the optimal regression coefficients learned in one hierarchy as feature weighting factors in the following hierarchy, and formulate a weighted sparse multi-task learning method. Lastly, we also take into account the distributional characteristics of samples per class and use clustering-induced subclass label vectors as target response values in our sparse regression model. In our experiments on the ADNI cohort, we performed both binary and multi-class classification tasks in AD/MCI diagnosis and showed the superiority of the proposed method by comparing with the state-of-the-art methods.


Treatment of gonorrhea: comparison of cefotaxime and penicillin.
Ninety-seven patients with 118 sites infected with Neisseria gonorrhoeae were treated with a single dose of either procaine penicillin G (4.8 x 10(6) U) or cefotoxime (1.0 g) intramuscularly. Only the penicillin group took 1 g of probenecid orally. The numbers of infected sites in each treatment group were as follows: penicillin-urethra, 37; rectum, 9; cervix, 8; and pharynx, 4; cefotaxime-urethra, 42; rectum, 9; cervix, 5; and pharynx, 4. The cure rates in each treatment group were 100%. No adverse reactions were noted in either group. beta-Lactamase-positive N. gonorrhoeae strains were not found. Ninety-five percent of clinical isolates were inhibited by less than or equal to 0.007 micrograms of cefotaxime and less than or equal to 0.25 micrograms of penicillin per ml. In this study cefotaxime was as effective as procaine penicillin in the treatment of uncomplicated gonorrhea.


Characterizing Body Image Distortion and Bodily Self-Plasticity in Anorexia Nervosa via Visuo-Tactile Stimulation in Virtual Reality.
We combined virtual reality and multisensory bodily illusion with the aim to characterize and reduce the perceptual (body overestimation) and the cognitive-emotional (body dissatisfaction) components of body image distortion (BID) in anorexia nervosa (AN). For each participant (20 anorexics, 20 healthy controls) we built personalized avatars that reproduced their own body size, shape, and verisimilar increases and losses of their original weight. Body overestimation and dissatisfaction were measured by asking participants to choose the avatar that best resembled their real and ideal body. Results show higher body dissatisfaction in AN, caused by the desire of a thinner body, and no body-size overestimation. Interpersonal multisensory stimulation (IMS) was then applied on the avatar reproducing participant's perceived body, and on the two avatars which reproduced increases and losses of 15% of it, all presented with a first-person perspective (1PP). Embodiment was stronger after synchronous IMS in both groups, but did not reduce BID in participants with AN. Interestingly, anorexics reported more negative emotions after embodying the fattest avatar, which scaled with symptoms severity. Overall, our findings suggest that the cognitive-emotional, more than the perceptual component of BID is severely altered in AN and that perspective (1PP vs. 3PP) from which a body is evaluated may play a crucial role. Future research and clinical trials might take advantage of virtual reality to reduce the emotional distress related to body dissatisfaction.


Statistics of random protein superpositions: p-values for pairwise structure alignment.
Quantification of statistical significance is essential for the interpretation of protein structural similarity. To address this, a random model for protein structure comparison was developed. Novelty of the model is threefold. First, a sample of random structure comparisons is restricted to molecules of the same size and shape as the superposition of interest. Second, careful selection of the sample and accurate modeling of shape allows approximation of the root mean square deviation (RMSD) distribution of random comparisons with a Nakagami probability density function. Third, through convolution, a second probability density function is obtained that describes the coordinate difference vector projections underlying the random distribution of RMSD. This last feature allows sampling random distributions of not only RMSD, but also any similarity score that depends on difference vector projections, such as GDT_TS score, TM score, and LiveBench 3D score. Probabilities estimated from the method correlate well with common measures of structural similarity, such as the Dali Z-score and the GDT_TS score. As a result, the p-value for a given superposition can be calculated using simple formulae depending on RMSD, radius of gyration, and thinnest molecular dimension. In addition to scoring structural similarity, p-values computed by this method can be applied to evaluation of homology modeling techniques, providing a statistically sound alternative to scores used in reference-independent evaluation of alignment quality.


Something in the way you move: dispersal pathways affect invasion success.
Biological invasions are caused by human-mediated extra-range dispersal and, unlike natural extra-range dispersal, are often the result of multiple introductions from multiple sources to multiple locations. The processes and opportunities that result in propagules moving from one area to another can be used more broadly to differentiate all types of extra-range dispersal. By examining key properties of dispersal pathways (notably propagule pressure, genetic diversity and the potential for simultaneous movement of coevolved species), the establishment and evolutionary trajectories of extra-range dispersal can be better understood. Moreover, elucidation of the mechanistic properties of dispersal pathways is crucial for scientists and managers who wish to assist, minimise or prevent future movements of organisms.


Medial-lateral organization of the orbitofrontal cortex.
Emerging evidence suggests that specific cognitive functions localize to different subregions of OFC, but the nature of these functional distinctions remains unclear. One prominent theory, derived from human neuroimaging, proposes that different stimulus valences are processed in separate orbital regions, with medial and lateral OFC processing positive and negative stimuli, respectively. Thus far, neurophysiology data have not supported this theory. We attempted to reconcile these accounts by recording neural activity from the full medial-lateral extent of the orbital surface in monkeys receiving rewards and punishments via gain or loss of secondary reinforcement. We found no convincing evidence for valence selectivity in any orbital region. Instead, we report differences between neurons in central OFC and those on the inferior-lateral orbital convexity, in that they encoded different sources of value information provided by the behavioral task. Neurons in inferior convexity encoded the value of external stimuli, whereas those in OFC encoded value information derived from the structure of the behavioral task. We interpret these results in light of recent theories of OFC function and propose that these distinctions, not valence selectivity, may shed light on a fundamental organizing principle for value processing in orbital cortex.


The Relationship of Chronological Age, Age at Injury, and Duration of Injury to Employment Status in Individuals with Spinal Cord Injury.
Employment status following spinal cord injury (SCI) has important implications for financial and psychosocial well-being. Several age-related variables-in particular chronological age, duration of SCI, and age at SCI onset-have been identified as being associated with employment among individuals with SCI. Cross-sectional investigations of this topic are complicated by methodological and statistical issues associated with aging and disability. The purpose of the current study was to examine the associations between three aging variables and employment status in individuals with SCI through a series of regression analyses. Six hundred twenty individuals with SCI completed a survey that included measures of demographic characteristics, pain, psychological functioning, physical functioning, fatigue, and sleep. The results indicated that chronological age and age at SCI onset were significant predictors of employment status. A significantly greater proportion of individuals aged 45-54 were employed compared to those aged 55-64 even after controlling for biopsychosocial variables. Additionally, there was a negative linear relationship between percent employed and age at SCI onset, and this relationship was not accounted for by the biopsychosocial variables. The analyses used in this study provide one method by which to disentangle the effects of different age-related variables on important SCI outcomes in cross-sectional research. Continued research in this area is needed to better understand age-related effects on employment status, which could be used to help maximize the quality of life in individuals with SCI.


Invisibility of Racism in the Global Neoliberal Era: Implications for Researching Racism in Healthcare.
This paper describes the difficulties of researching racism in healthcare contexts as part of the wider issue of neoliberal reforms in welfare states in the age of global migration. In trying to understand the contradiction of a phenomenon that is historical and strongly felt by individuals and yet widely denied by both institutions and individuals, we consider the current political and socioeconomic context of healthcare provision. Despite decades of legislation against racism, its presence persists in healthcare settings, but data on these experiences is rarely gathered in Europe. National systems of healthcare provision have been subject to neoliberal reforms, where among others, cheaper forms of labor are sought to reduce the cost of producing healthcare, while the availability of services is rationed to contain demand. The restriction both on provision of and access to welfare, including healthcare, is unpopular among national populations. However, the explanations for restricted access to healthcare are assumed to be located outside the national context with immigrants being blamed. Even as migrants are used as a source of cheap labor in healthcare and other welfare sectors, the arrival of immigrants has been held responsible for restricted access to healthcare and welfare in general. One implication of (im)migration being blamed for healthcare restrictions, while racism is held to be a problem of the past, is the silencing of experiences of racism, which has dire consequences for ethnic minority populations. The implications of racism as a form of inequality within healthcare and the circumstances of researching racism in healthcare and its implication for the sociology of health in Sweden are described.


Intensive care readmission: a contemporary review of the literature.
ICU readmissions are a commonly used quality measure but despite decades of research, these adverse events continue to occur. Of particular concern is that readmitted patients have much worse prognoses than those not readmitted. In recent years new clinical service roles have evolved to assist ward staff with the care of acutely ill patients, such as those discharged from ICU. Given the recent emergence of these service roles, a review of contemporary ICU readmission studies was warranted to determine their impact on this adverse event. Reviewed studies indicated the incidence of readmissions and outcomes of these patients have changed little in recent years. Few studies mentioned whether clinical service roles existed to support ward staff caring for patients recently discharged from ICU. Future research needs to focus on identifying modifiable factors in care processes to reduce the incidence and outcomes of this adverse event and to determine how clinical service roles can best help prevent its occurrence.


New therapeutic approach to suppress castration-resistant prostate cancer using ASC-J9 via targeting androgen receptor in selective prostate cells.
Using androgen receptor (AR) knockout mice to determine AR functions in selective prostate cancer (PCa) cells, we determined that AR might play differential roles in various cell types, either to promote or suppress PCa development/progression. These observations partially explain the failure of current androgen deprivation therapy (ADT) to reduce/prevent androgen binding to AR in every cell. Herein, we identified the AR degradation enhancer ASC-J9, which selectively degrades AR protein via interruption of the AR-AR selective coregulator interaction. Such selective interruption could, therefore, suppress AR-mediated PCa growth in the androgen-sensitive stage before ADT and in the castration-resistant stage after ADT. Mechanistic dissection suggested that ASC-J9 could activate the proteasome-dependent pathway to promote AR degradation through the enhanced association of AR-Mdm2 complex. The consequences of ASC-J9-promoted AR degradation included reduced androgen binding to AR, AR N-C terminal interaction, and AR nuclear translocation. Such inhibitory regulation could then result in suppression of AR transactivation and AR-mediated cell growth in eight different mouse models, including intact or castrated nude mice xenografted with androgen-sensitive LNCaP cells or androgen-insensitive C81 cells and castrated nude mice xenografted with castration-resistant C4-2 and CWR22Rv1 cells, and TRAMP and Pten(+/-) mice. These results demonstrate that ASC-J9 could serve as an AR degradation enhancer that effectively suppresses PCa development/progression in the androgen-sensitive and castration-resistant stages.


Global expression analysis of the brown alga Ectocarpus siliculosus (Phaeophyceae) reveals large-scale reprogramming of the transcriptome in response to abiotic stress.
Brown algae (Phaeophyceae) are phylogenetically distant from red and green algae and an important component of the coastal ecosystem. They have developed unique mechanisms that allow them to inhabit the intertidal zone, an environment with high levels of abiotic stress. Ectocarpus siliculosus is being established as a genetic and genomic model for the brown algal lineage, but little is known about its response to abiotic stress. Here we examine the transcriptomic changes that occur during the short-term acclimation of E. siliculosus to three different abiotic stress conditions (hyposaline, hypersaline and oxidative stress). Our results show that almost 70% of the expressed genes are regulated in response to at least one of these stressors. Although there are several common elements with terrestrial plants, such as repression of growth-related genes, switching from primary production to protein and nutrient recycling processes, and induction of genes involved in vesicular trafficking, many of the stress-regulated genes are either not known to respond to stress in other organisms or are have been found exclusively in E. siliculosus. This first large-scale transcriptomic study of a brown alga demonstrates that, unlike terrestrial plants, E. siliculosus undergoes extensive reprogramming of its transcriptome during the acclimation to mild abiotic stress. We identify several new genes and pathways with a putative function in the stress response and thus pave the way for more detailed investigations of the mechanisms underlying the stress tolerance of brown algae.


Structural insight into the differential interactions between the DNA mimic protein SAUGI and two gamma herpesvirus uracil-DNA glycosylases.
Uracil-DNA glycosylases (UDGs) are conserved DNA-repair enzymes that can be found in many species, including herpesviruses. Since they play crucial roles for efficient viral DNA replication in herpesviruses, they have been considered as potential antiviral targets. In our previous work, Staphylococcus aureus SAUGI was identified as a DNA mimic protein that targets UDGs from S. aureus, human, Herpes simplex virus (HSV) and Epstein-Barr virus (EBV). Interestingly, SAUGI has the strongest inhibitory effects with EBVUDG. Here, we determined complex structures of SAUGI with EBVUDG and another γ-herpesvirus UDG from Kaposi's sarcoma-associated herpesvirus (KSHVUDG), which SAUGI fails to effectively inhibit. Structural analysis of the SAUGI/EBVUDG complex suggests that the additional interaction between SAUGI and the leucine loop may explain why SAUGI shows the highest binding capacity with EBVUDG. In contrast, SAUGI appears to make only partial contacts with the key components responsible for the compression and stabilization of the DNA backbone in the leucine loop extension of KSHVUDG. The findings in this study provide a molecular explanation for the differential inhibitory effects and binding strengths that SAUGI has on these two UDGs, and the structural basis of the differences should be helpful in developing inhibitors that would interfere with viral DNA replication.


Unrevealing sequence and structural features of novel coronavirus using in silico approaches: The main protease as molecular target.
Direct-acting antivirals are effective tools to control viral infections. SARS-CoV-2 is a coronavirus associated with the epidemiological outbreak in late 2019. Previous reports showed that HIV-1 protease inhibitors could block SARS-CoV main protease. Based on that and using an in silico approach, we evaluated SARS-CoV-2 main protease as a target for HIV-1 protease inhibitors to reveal the structural features related to their antiviral effect. Our results showed that several HIV inhibitors such as lopinavir, ritonavir, and saquinavir produce strong interaction with the active site of SARS-CoV-2 main protease. Furthermore, broad library protease inhibitors obtained from PubChem and ZINC (www.zinc.docking.org) were evaluated. Our analysis revealed 20 compounds that could be clustered into three groups based on their chemical features. Then, these structures could serve as leading compounds to develop a series of derivatives optimizing their activity against SARS-CoV-2 and other coronaviruses. Altogether, the results presented in this work contribute to gain a deep understanding of the molecular pharmacology of SARS-CoV-2 treatment and validate the use of protease inhibitors against SARS-CoV-2.


Cortical thickness and brain volumetric analysis in body dysmorphic disorder.
Individuals with body dysmorphic disorder (BDD) suffer from preoccupations with perceived defects in physical appearance, causing severe distress and disability. Although BDD affects 1-2% of the population, the neurobiology is not understood. Discrepant results in previous volumetric studies may be due to small sample sizes, and no study has investigated cortical thickness in BDD. The current study is the largest neuroimaging analysis of BDD. Participants included 49 medication-free, right-handed individuals with DSM-IV BDD and 44 healthy controls matched by age, sex, and education. Using high-resolution T1-weighted magnetic resonance imaging, we computed vertex-wise gray matter (GM) thickness on the cortical surface and GM volume using voxel-based morphometry. We also computed volumes in cortical and subcortical regions of interest. In addition to group comparisons, we investigated associations with symptom severity, insight, and anxiety within the BDD group. In BDD, greater anxiety was significantly associated with thinner GM in the left superior temporal cortex and greater GM volume in the right caudate nucleus. There were no significant differences in cortical thickness, GM volume, or volumes in regions of interest between BDD and control subjects. Subtle associations with clinical symptoms may characterize brain morphometric patterns in BDD, rather than large group differences in brain structure.


Metaplastic Carcinoma of the Breast: Analysis of 38 Cases from a Single Institute.
To evaluate the pathological and radiological features, hormone profiles, surgery and treatment methods of metaplastic breast carcinoma cases diagnosed at our center in the light of current literature. A total of 38 metaplastic breast cancer cases diagnosed between 2006-2018 at our center were included in the study. The patients were evaluated in terms of age, tumor size, localization, histological grade, hormone profiles (ER, PR, Her2-neu), American Joint Committee on Cancer (AJCC) Tumor, Lymph node status, Metastases (TNM) stage, progression, survival, radiological features, types of surgery and therapy modalities (chemotherapy and / or radiotherapy). The age of the patients ranged between 32 and 95 years. Pathological evaluation of cases showed that 14 were pure epithelial (IC-NST + squamous cell carcinoma) and 24 were metaplastic carcinomas with mesenchymal differentiation. Ductal carcinoma in situ (DCIS) was accompanying an invasive component in twenty cases. Seventeen patients had lymph node metastasis. Twelve patients developed distant metastasis. Thirty patients were triple negative for hormone receptors. The mean follow-up period of the patients was 34 months. The estimated life expectancy was 116 months. All of the patients received chemotherapy and 28 patients received adjuvant radiotherapy. There was no correlation between tumor size and lymph node or distant metastasis in our series. Our findings are consistent with the literature. Metaplastic breast carcinoma is a rare entity among breast carcinomas. Metaplastic carcinomas of the breast draw attention with the differences in their clinical course and the radiological and pathological heterogeneity.


The gut microbiome in psychiatry: A primer for clinicians.
Research in the past decade has shown that variations in the gut microbiome may influence behavior, and vice versa. As such, interest in the role of the gut microbiome in psychiatric conditions has drawn immense interest. This is evidenced by the recent surge in published studies examining microbial dysbiosis in clinical psychiatric populations, particularly autism spectrum disorder and depression. However, critical examination of these studies reveals methodological flaws in design and execution, suggesting that they may not be held to the same standards as other bodies of clinical research. Given the complex nature of the gut microbiome, this narrative review attempts to clarify concepts critical to effectively examine its potential role in psychopathology to appropriately inform mental health researchers. More specifically, the numerous variables known to affect the gut microbiome are discussed, including inflammation, diet, weight, and medications. A comprehensive review of the extant microbiome literature in clinical psychiatric populations is also provided, in addition to clinical implications and suggestions for future directions of research. Although there is a clear need for additional studies to elucidate the gut microbiome's role in psychiatric disorders, there is an even greater need for well-designed, appropriately controlled studies to truly impact the field.


A comparison of the effects of problem-based learning and lecturing on the development of students' critical thinking.
Educational approaches are thought to have facilitative or hindering effects on students' critical thinking development. The aim of this study was to compare the effects of problem-based learning (PBL) and lecturing approaches on the development of students' critical thinking. All 79 Year 1 undergraduate nursing students at a university in Hong Kong were randomly assigned to 1 of 2 parallel courses delivered by either PBL (n = 40) or lecturing (n = 39) over 1 academic year. The primary outcome measure was students' critical thinking disposition as measured by the California Critical Thinking Disposition Inventory (CCTDI). Individual interviews were also conducted to elicit the students' perceptions of their learning experience. Data were collected at 4 timepoints spanning 3 years. The overall CCTDI and subscale scores for the PBL group were not significantly different from those of the lecture group at the first timepoint (pretest). Compared with lecture students, PBL students showed significantly greater improvement in overall CCTDI (P = 0.0048), Truthseeking (P = 0.0008), Analyticity (P = 0.0368) and Critical Thinking Self-confidence (P = 0.0342) subscale scores from the first to the second timepoints; in overall CCTDI (P = 0.0083), Truthseeking (P = 0.0090) and Analyticity (P = 0.0354) subscale scores from the first to the third timepoints; and in Truthseeking (P = 0.0173) and Systematicity (P = 0.0440) subscale scores from the first to the fourth timepoints. There were significant differences in the development of students' critical thinking dispositions between those who undertook the PBL and lecture courses, respectively.


Targeted association analysis identified japonica rice varieties achieving Na(+)/K (+) homeostasis without the allelic make-up of the salt tolerant indica variety Nona Bokra.
During the last decade, a large number of QTLs and candidate genes for rice tolerance to salinity have been reported. Using 124 SNP and 52 SSR markers, we targeted 14 QTLs and 65 candidate genes for association mapping within the European Rice Core collection (ERCC) comprising 180 japonica accessions. Significant differences in phenotypic response to salinity were observed. Nineteen distinct loci significantly associated with one or more phenotypic response traits were detected. Linkage disequilibrium between these loci was extremely low, indicating a random distribution of favourable alleles in the ERCC. Analysis of the function of these loci indicated that all major tolerance mechanisms were present in the ERCC although the useful level of expression of the different mechanisms was scattered among different accessions. Under moderate salinity stress some accessions achieved the same level of control of Na(+) concentration and Na(+)/K(+) equilibrium as the indica reference variety for salinity tolerance Nona Bokra, although without sharing the same alleles at several loci associated with Na(+) concentration. This suggests (a) differences between indica and japonica subspecies in the effect of QTLs and genes involved in salinity tolerance and (b) further potential for the improvement of tolerance to salinity above the tolerance level of Nona Bokra, provided the underlying mechanisms are complementary at the whole plant level. No accession carried all favourable alleles, or showed the best phenotypic responses for all traits measured. At least nine accessions were needed to assemble the favourable alleles and all the best phenotypic responses. An effective strategy for the accumulation of the favourable alleles would be marker-assisted population improvement.


Picosecond resolution in scanning tunneling microscopy.
A method has been developed for performing fast time-resolved experiments with a scanning tunneling microscope. The method uses the intrinsic nonlinearity in the microscope's current versus voltage characteristics to resolve optically generated transient signals on picosecond time scales. The ability to combine the spatial resolution of tunneling microscopy with the time resolution of ultrafast optics yields a powerful tool for the investigation of dynamic phenomena on the atomic scale.


Evaluating statistical methods used to estimate the number of postsynaptic receptors.
Calcium levels in spines play a significant role in determining the sign and magnitude of synaptic plasticity. The magnitude of calcium influx into spines is highly dependent on influx through N-methyl D-aspartate (NMDA) receptors, and therefore depends on the number of postsynaptic NMDA receptors in each spine. We have calculated previously how the number of postsynaptic NMDA receptors determines the mean and variance of calcium transients in the postsynaptic density, and how this alters the shape of plasticity curves. However, the number of postsynaptic NMDA receptors in the postsynaptic density is not well known. Anatomical methods for estimating the number of NMDA receptors produce estimates that are very different than those produced by physiological techniques. The physiological techniques are based on the statistics of synaptic transmission and it is difficult to experimentally estimate their precision. In this paper we use stochastic simulations in order to test the validity of a physiological estimation technique based on failure analysis. We find that the method is likely to underestimate the number of postsynaptic NMDA receptors, explain the source of the error, and re-derive a more precise estimation technique. We also show that the original failure analysis as well as our improved formulas are not robust to small estimation errors in key parameters.


Decisionmaking in regional health planning agencies.
Health Systems Agencies (HSAs), the new regional health planning agencies established by the National Health Planning and Resources Development Act of 1974, have as their major goals quality, accessibility, continuity and cost containment. One of the tools for cost containment available to HSAs is their active participation in the statewide certificate-of-need (CON) program. To help anticipate how HSAs may play their CON roles, this article reviews the CON decisionmaking processes of two Comprehensive Health Planning (CHP) agencies, the predecessors of HSAs. While most CHPs lacked specific health plans and powers. the two California agencies examined have had both regional plans and roles in CON programs comparable to the present HSAs'. Our focus is on the decisionmaking process rather than the effectiveness of CON programs. A simple descriptive model is developed based on the assumptions that the planners seek satisfactory, rather than optimal solutions and that the data available are extremely limited. The cases analyzed suggest that, while most projects exceed the bed need standards, institutions were almost always able to demonstrate a need through various special circumstances. Once a need has been shown, the agency was able to deny an application only if a better alternative were available. This study suggests two approaches to improving regional health planning decisionmaking. First, identified needs can be better met either by increasing the agency's ability to generate alternatives or by encouraging health facilities to submit competing applications. Second, a regional health care budget would force the agency to make choices among competing needs.


Characterization of the B Cell Transcriptome Bound by RNA-Binding Proteins with iCLIP.
Posttranscriptional regulation of gene expression shapes the B cell transcriptome and controls messenger RNA (mRNA) translation into protein. Recent reports have highlighted the importance of RNA binding proteins (RBPs) for mRNA splicing, subcellular location, stability, and translation during B lymphocyte development, activation, and differentiation. Here we describe individual-nucleotide resolution UV cross-linking and immunoprecipitation (iCLIP) in primary lymphocytes, a method that maps RNA-protein interactions in a genome-wide scale allowing mechanistic analysis of RBP function. We discuss the latest improvements in iCLIP technology and provide some examples of how integration of the RNA-protein interactome with other high-throughput mRNA sequencing methodologies uncovers the important role of RBP-mediated RNA regulation in key biological cell processes.


Probing the reaction pathways of DL-proline on TiO2 (001) single crystal surfaces.
The reaction of DL-Proline on O2-annealed (stoichiometric) and O-defected (sub-stoichiometric) TiO2 (001) single-crystal surfaces has been investigated. This is of significance in trying to understand the concept of how biomolecules interact with the surfaces of biomedical implants (molecular recognition). On an O2-annealed TiO2 surface, proline is found to largely adsorb then desorb intact at approximately 350 K. DFT (B3LYP) calculations of proline bound to a Ti(OH)4 cluster suggest a binding through the carboxylate functional group rather than through the NH group of the ring. In contrast, proline reaction was considerably different on the O-defected surface. First, proline was further stabilized, evidenced by a shift of its desorption temperature (during temperature-programmed desorption) to approximately 530 K. Along with proline desorption, two distinctive sets of reaction processes occurred at 530 and 630 K, respectively. The first pathway (alpha) at 530 K shows desorption of large amounts of m/e 55 (attributed to 1-azetine) and m/e 42 (attributed to ketene). At still higher temperature, 630 K, a pathway (beta) dominated by the appearance of low masses, mainly m/e 28, 27, and 26, is seen. These masses are tentatively attributed to desorption of HCN, ethylene, and/or acetylene as they represent the logical further decomposition of the different fragments of proline.


Molecular characterization of Citrus tatter leaf virus historically associated with Meyer lemon trees: complete genome sequence and development of biologically active in vitro transcripts.
Citrus tatter leaf virus isolated from Meyer lemon trees (CTLV-ML) from California and Florida induces bud union incompatibility of citrus trees grafted on the widely used trifoliate and trifoliate hybrid rootstocks. The complete genome sequence of CTLV-ML was determined to be 6,495 nucleotides (nts), with two overlapping open reading frames (ORFs) and a poly (A) tail at the 3' end. The genome organization is similar to other capilloviruses, with ORF1 (nts 37 to 6,354) encoding a putative 242-kDa polyprotein which contains replication-associated domains plus a coat protein (CP), and ORF2 (nts 4,788 to 5,750), which is located within ORF1 in a different reading frame and encodes a putative movement protein. Although the proteins encoded by CTLV-ML possesses 84 to 96% amino acid sequence identity with strains of Apple stem grooving virus (ASGV), we observed two strikingly different regions in ORF1: variable region I (amino acids 532 to 570) and variable region II (amino acids 1,583 to 1,868), with only 15 to 18 and 56 to 62% identities, respectively, with the corresponding regions of ASGV strains. Conditions for a herbaceous systemic assay host were optimized in which the wild-type virus induced systemic infection in Phaseolus vulgaris cv. Light Red Kidney (LRK) bean plants at 19 or 22 degrees C but not at higher temperatures. In vitro transcripts generated from full-length cDNA clones induced systemic symptoms on LRK bean plants similar to that of the wild-type virus. Replication of the recombinant virus was confirmed by hybridization of a 5' positive-stranded RNA-specific probe to a genome-sized RNA and by reverse-transcription polymerase chain reaction.


Evidence for altered LRP/RAGE expression in Alzheimer lesion pathogenesis.
There is significant evidence to suggest that a damaged or dysfunctional blood-brain barrier (BBB) may contribute to the pathogenesis of Alzheimer's disease (AD) lesions. Lipoprotein receptor-related protein (LRP-1) and receptor for advanced glycation end products (RAGE) are known to be important (BBB) capillary transport proteins. Altered expression of either of these capillary endothelial LRP-1 and RAGE receptor proteins could indicate a dysfunction of the BBB and its transport regulation of beta-amyloid (Abeta). Cortical samples from the superior temporal (ST) and calcarine occipital (COC) cortices of ten confirmed AD brains and ten comparison group (CG) brains were examined. The densities of neurofibrillary tangles (NFTs), senile plaques (SPs) and LRP-1 and RAGE positive capillaries were recorded and statistically analyzed. There was a statistically significant difference between AD and CG cases and the densities of LRP-1 and RAGE positive capillaries, the AD cases demonstrating the greater numbers. Further, in AD brains there were significant negative correlations between the Abeta burden of SPs and both LRP-1 and RAGE-positive capillaries [p < .001]. Additionally, there was a strong positive correlation between LRP-1 and RAGE capillaries in AD brains [p < .001]. These results suggest that alterations in the LRP-1 and RAGE mediated transport of Abeta take place in AD brains in lesion prone regions and may therefore contribute to SP lesion pathogenesis.


Assessment of copy number variation using the Illumina Infinium 1M SNP-array: a comparison of methodological approaches in the Spanish Bladder Cancer/EPICURO study.
High-throughput single nucleotide polymorphism (SNP)-array technologies allow to investigate copy number variants (CNVs) in genome-wide scans and specific calling algorithms have been developed to determine CNV location and copy number. We report the results of a reliability analysis comparing data from 96 pairs of samples processed with CNVpartition, PennCNV, and QuantiSNP for Infinium Illumina Human 1Million probe chip data. We also performed a validity assessment with multiplex ligation-dependent probe amplification (MLPA) as a reference standard. The number of CNVs per individual varied according to the calling algorithm. Higher numbers of CNVs were detected in saliva than in blood DNA samples regardless of the algorithm used. All algorithms presented low agreement with mean Kappa Index (KI) <66. PennCNV was the most reliable algorithm (KI(w=) 98.96) when assessing the number of copies. The agreement observed in detecting CNV was higher in blood than in saliva samples. When comparing to MLPA, all algorithms identified poorly known copy aberrations (sensitivity = 0.19-0.28). In contrast, specificity was very high (0.97-0.99). Once a CNV was detected, the number of copies was truly assessed (sensitivity >0.62). Our results indicate that the current calling algorithms should be improved for high performance CNV analysis in genome-wide scans. Further refinement is required to assess CNVs as risk factors in complex diseases.


Production and purification of lentiviral vectors generated in 293T suspension cells with baculoviral vectors.
Lentivirus can be engineered to be a highly potent vector for gene therapy applications. However, generation of clinical grade vectors in enough quantities for therapeutic use is still troublesome and limits the preclinical and clinical experiments. As a first step to solve this unmet need we recently introduced a baculovirus-based production system for lentiviral vector (LV) production using adherent cells. Herein, we have adapted and optimized the production of these vectors to a suspension cell culture system using recombinant baculoviruses delivering all elements required for a safe latest generation LV preparation. High-titer LV stocks were achieved in 293T cells grown in suspension. Produced viruses were accurately characterized and the functionality was also tested in vivo. Produced viruses were compared with viruses produced by calcium phosphate transfection method in adherent cells and polyethylenimine transfection method in suspension cells. Furthermore, a scalable and cost-effective capture purification step was developed based on a diethylaminoethyl monolithic column capable of removing most of the baculoviruses from the LV pool with 65% recovery.


Sterically induced reductive linkage of iron polypnictides with bulky lanthanide complexes by ring-opening of THF.
Reduction of [Cp*Fe(η5-E5)] (E = P, As) with divalent lanthanide reagents usually leads to reduction of [Cp*Fe(η5-E5)] followed by a Ln-E bond formation. In contrast, by using the sterically encumbered reagent [(DippForm)2Sm(thf)2] (DippForm = {(2,6-iPr2C6H3)NC(H)[double bond, length as m-dash]N(2,6-iPr2C6H3)}-), ring-opening of thf and reduction of the polypnictide is observed. This leads to two new 3d/4f polyphosphide or polyarsenide complexes [(DippForm)2Sm(Cp*Fe)E5{(CH2)4O}{(DippForm)2Sm(thf)}], in which [(DippForm)2Sm(thf)2] and [Cp*Fe(η5-E5)] are linked by a ring-opened thf molecule and no Ln-E bond formation is observed.


Biomechanical association of dental and temporomandibular pathology in a medieval Nubian population.
An analysis of the relationship between oral pathology and degenerative change at the temporomandibular joint (TMJ) was undertaken on an archaeological sample of 122 adult crania from the Medieval site of Kulubnarti in Sudanese Nubia. The crania were sorted into 2 groups: those demonstrating clearly visible bony changes at the joint (TMJ+) and those without visible change (TMJ-). These groups were compared according to 1) age; 2) sex; 3) active dental pathologies (abscesses, caries, partial socket resorption); 4) tooth loss with complete socket resorption; and 5) dental attrition. No statistically significant association was evident between degenerative change at the TMJ and age, active dental pathologies, or dental attrition; however, sex differences and posterior tooth loss with complete socket resorption revealed a significant correspondence to degenerative TMJ changes. Both of these factors agree with the clinical literature and with biomechanical models (most notably that of Hylander) based upon modern populations. Furthermore, the results support the contention that paleopathological conditions can be analyzed from a clinical and functional biomechanical perspective.


A potential tool for high-resolution monitoring of ocean acidification.
Current anthropogenic carbon dioxide emissions generate besides global warming unprecedented acidification rates of the oceans. Recent evidence indicates the possibility that ocean acidification and low oceanic pH may be a major reason for several mass extinctions in the past. However, a major bottleneck for research on ocean acidification is long-term monitoring and the collection of consistent high-resolution pH measurements. This study presents a low-power (<1 W) small sample volume (25 μL) semiconductor based fluorescence method for real-time ship-board pH measurements at high temporal and spatial resolution (approximately 15 s and 100 m between samples). A 405 nm light emitting diode and the blue and green channels from a digital camera was used for swift detection of fluorescence from the pH sensitive dye 6,8-Dihydroxypyrene-1,3-disulfonic acid in real-time. Main principles were demonstrated by automated continuous measurements of pH in the surface water across the Baltic Sea and the Kattegat region with a large range in salinity (~3-30) and temperature (~0-25°C). Ship-board precision of salinity and temperature adjusted pH measurements were estimated as low as 0.0001 pH units.


Very high-resolution spectroscopy for extremely large telescopes using pupil slicing and adaptive optics.
Under seeing limited conditions very high resolution spectroscopy becomes very difficult for extremely large telescopes (ELTs). Using adaptive optics (AO) the stellar image size decreases proportional with the telescope diameter. This makes the spectrograph optics and hence its resolution independent of the telescope diameter. However AO for use with ELTs at visible wavelengths require deformable mirrors with many elements. Those are not likely to be available for quite some time. We propose to use the pupil slicing technique to create a number of sub-pupils each of which having its own deformable mirror. The images from all sub-pupils are combined incoherently with a diameter corresponding to the diffraction limit of the sub-pupil. The technique is referred to as "Pupil Slicing Adaptive Optics" or PSAO.


Estimation of feed crude protein concentration and rumen degradability by Fourier-transform infrared spectroscopy.
Currently, rapid methods are needed for feed analysis. This study examined the potential of Fourier-transform infrared (FTIR) spectroscopy to predict the nutritional value of a wide range of feeds for ruminants, as an alternative to the in situ technique. Moreover, we investigated whether universal equations could be developed that would allow the low-cost determination of crude protein (CP) concentrations and their kinetics of degradation into the rumen. Protein nutritional values of 663 samples comprising 80 different feed types were determined in terms of concentrations of CP, water-soluble CP (CP(WS)), total-tract mobile bag CP digestibility (CP(TTD)), and in situ CP degradability, including the rumen soluble fraction (CP(A)), the degradable but not soluble fraction (CP(B)), rate of CP(B) degradation (CP(C)), effective degradability (CP(ED)), and potential degradability (CPPD). Infrared spectra of dry samples were collected by attenuated total reflectance from 4000 to 600 cm(-1). Models were developed by partial least squares (PLS) regression in a randomly selected subset of samples, and the precision of the equations was confirmed by using an external validation set. Analysis by FTIR spectroscopy was sufficiently sensitive to allow the accurate prediction of sample CP concentration (R(2)=0.92) and to classify feeds according to their CPWS concentrations using universal models (R(2)=0.78) that included all sample types. Moreover, substantial improvements in predictions were observed when samples were subdivided in groups. Models for forages led to accurate predictions of CP(WS) and fractions CP(A) and CP(B) (R(2)>0.83), whereas models for CP(TTD) and CP(ED) could be used for screening purposes (R(2)>0.67). This study showed that models for protein-rich concentrates alone could also be used for screening according to the feed concentrations of CP(WS), CP(TTD), CP(ED), CP(A), and CP(B), but models for energy-rich concentrates gave relatively poor predictions. The general difficulty observed in predicting CP(C) is because of a low correlation between FTIR spectra and the kinetics of CP degradation, which may be the result of large variation in the reference method (i.e., in situ degradation studies) and perhaps also because of the presence of compounds that can modify the CP degradation pattern in the rumen. In conclusion, FTIR spectroscopy should be considered as a low-cost alternative in the feed evaluation industry.


Precursor-centric genome-mining approach for lasso peptide discovery.
Lasso peptides are a class of ribosomally synthesized posttranslationally modified natural products found in bacteria. Currently known lasso peptides have a diverse set of pharmacologically relevant activities, including inhibition of bacterial growth, receptor antagonism, and enzyme inhibition. The biosynthesis of lasso peptides is specified by a cluster of three genes encoding a precursor protein and two enzymes. Here we develop a unique genome-mining algorithm to identify lasso peptide gene clusters in prokaryotes. Our approach involves pattern matching to a small number of conserved amino acids in precursor proteins, and thus allows for a more global survey of lasso peptide gene clusters than does homology-based genome mining. Of more than 3,000 currently sequenced prokaryotic genomes, we found 76 organisms that are putative lasso peptide producers. These organisms span nine bacterial phyla and an archaeal phylum. To provide validation of the genome-mining method, we focused on a single lasso peptide predicted to be produced by the freshwater bacterium Asticcacaulis excentricus. Heterologous expression of an engineered, minimal gene cluster in Escherichia coli led to the production of a unique lasso peptide, astexin-1. At 23 aa, astexin-1 is the largest lasso peptide isolated to date. It is also highly polar, in contrast to many lasso peptides that are primarily hydrophobic. Astexin-1 has modest antimicrobial activity against its phylogenetic relative Caulobacter crescentus. The solution structure of astexin-1 was determined revealing a unique topology that is stabilized by hydrogen bonding between segments of the peptide.


Reduction of unnecessary intravenous catheter use. Internal medicine house staff participate in a successful quality improvement project.
We had previously observed that a large proportion of peripheral intravenous (i.v.) catheters placed in patients on a regular medical ward at our hospital were unnecessary. We conducted the current study to assess the effect of a quality improvement project led by medicine house staff on the prevalence of unnecessary peripheral i.v. catheters (those without any therapeutic use, referred to as idle). All patients on four regular-care medical wards of a large university-affiliated veterans hospital were included in the study. The proportion of i.v. catheter episodes in which catheters were idle 2 or more consecutive days (idle episodes) and the proportion of patients exposed to an idle catheter episode were determined by direct observation, chart review, and patient interview before and after a multidisciplinary quality improvement task force defined guidelines for appropriate i.v. catheter use and made recommendations for hospital policy changes related to i.v. catheter use. The proportion of all i.v. catheter episodes that were idle catheter episodes decreased significantly after the intervention (42% before vs 29% after, P < .01), as did the proportion of patients with an i.v. catheter who had at least one idle i.v. catheter episode (43% vs 27%, P < .001). This quality improvement effort successfully reduced unnecessary i.v. catheter use. We suspect that house-staff involvement in the intervention was critical. We encourage other academic medical centers to involve house staff in quality improvement activities to improve patient care and to enhance the education of house staff regarding quality improvement processes.


Mechanism of neutralization by the broadly neutralizing HIV-1 monoclonal antibody VRC01.
The structure of VRC01 in complex with the HIV-1 gp120 core reveals that this broadly neutralizing CD4 binding site (CD4bs) antibody partially mimics the interaction of the primary virus receptor, CD4, with gp120. Here, we extended the investigation of the VRC01-gp120 core interaction to the biologically relevant viral spike to better understand the mechanism of VRC01-mediated neutralization and to define viral elements associated with neutralization resistance. In contrast to the interaction of CD4 or the CD4bs monoclonal antibody (MAb) b12 with the HIV-1 envelope glycoprotein (Env), occlusion of the VRC01 epitope by quaternary constraints was not a major factor limiting neutralization. Mutagenesis studies indicated that VRC01 contacts within the gp120 loop D, the CD4 binding loop, and the V5 region were necessary for optimal VRC01 neutralization, as suggested by the crystal structure. In contrast to interactions with the soluble gp120 monomer, VRC01 interaction with the native viral spike did not occur in a CD4-like manner; VRC01 did not induce gp120 shedding from the Env spike or enhance gp41 membrane proximal external region (MPER)-directed antibody binding to the Env spike. Finally, VRC01 did not display significant reactivity with human antigens, boding well for potential in vivo applications. The data indicate that VRC01 interacts with gp120 in the context of the functional spike in a manner distinct from that of CD4. It achieves potent neutralization by precisely targeting the CD4bs without requiring alterations of Env spike configuration and by avoiding steric constraints imposed by the quaternary structure of the functional Env spike.


Genome-wide association of functional traits linked with Campylobacter jejuni survival from farm to fork.
Campylobacter jejuni is a major cause of bacterial gastroenteritis worldwide, primarily associated with the consumption of contaminated poultry. C. jejuni lineages vary in host range and prevalence in human infection, suggesting differences in survival throughout the poultry processing chain. From 7343 MLST-characterised isolates, we sequenced 600 C. jejuni and C. coli isolates from various stages of poultry processing and clinical cases. A genome-wide association study (GWAS) in C. jejuni ST-21 and ST-45 complexes identified genetic elements over-represented in clinical isolates that increased in frequency throughout the poultry processing chain. Disease-associated SNPs were distinct in these complexes, sometimes organised in haplotype blocks. The function of genes containing associated elements was investigated, demonstrating roles for cj1377c in formate metabolism, nuoK in aerobic survival and oxidative respiration, and cj1368-70 in nucleotide salvage. This work demonstrates the utility of GWAS for investigating transmission in natural zoonotic pathogen populations and provides evidence that major C. jejuni lineages have distinct genotypes associated with survival, within the host specific niche, from farm to fork.


Zebrafish bioassay-guided microfractionation identifies anticonvulsant steroid glycosides from the Philippine medicinal plant Solanum torvum.
Medicinal plants used for the treatment of epilepsy are potentially a valuable source of novel antiepileptic small molecules. To identify anticonvulsant secondary metabolites, we performed an in vivo, zebrafish-based screen of medicinal plants used in Southeast Asia for the treatment of seizures. Solanum torvum Sw. (Solanaceae) was identified as having significant anticonvulsant activity in zebrafish larvae with seizures induced by the GABAA antagonist pentylenetetrazol (PTZ). This finding correlates well with the ethnomedical use of this plant in the Philippines, where a water decoction of S. torvum leaves is used to treat epileptic seizures. HPLC microfractionation of the bioactive crude extract, in combination with the in vivo zebrafish seizure assay, enabled the rapid localization of several bioactive compounds that were partially identified online by UHPLC-TOF-MS as steroid glycosides. Targeted isolation of the active constituents from the methanolic extract enabled the complete de novo structure identification of the six main bioactive compounds that were also present in the traditional preparation. To partially mimic the in vivo metabolism of these triterpene glycosides, their common aglycone was generated by acid hydrolysis. The isolated molecules exhibited significant anticonvulsant activity in zebrafish seizure assays. These results underscore the potential of zebrafish bioassay-guided microfractionation to rapidly identify novel bioactive small molecules of natural origin.


Consensus paper: Decoding the Contributions of the Cerebellum as a Time Machine. From Neurons to Clinical Applications.
Time perception is an essential element of conscious and subconscious experience, coordinating our perception and interaction with the surrounding environment. In recent years, major technological advances in the field of neuroscience have helped foster new insights into the processing of temporal information, including extending our knowledge of the role of the cerebellum as one of the key nodes in the brain for this function. This consensus paper provides a state-of-the-art picture from the experts in the field of the cerebellar research on a variety of crucial issues related to temporal processing, drawing on recent anatomical, neurophysiological, behavioral, and clinical research.The cerebellar granular layer appears especially well-suited for timing operations required to confer millisecond precision for cerebellar computations. This may be most evident in the manner the cerebellum controls the duration of the timing of agonist-antagonist EMG bursts associated with fast goal-directed voluntary movements. In concert with adaptive processes, interactions within the cerebellar cortex are sufficient to support sub-second timing. However, supra-second timing seems to require cortical and basal ganglia networks, perhaps operating in concert with cerebellum. Additionally, sensory information such as an unexpected stimulus can be forwarded to the cerebellum via the climbing fiber system, providing a temporally constrained mechanism to adjust ongoing behavior and modify future processing. Patients with cerebellar disorders exhibit impairments on a range of tasks that require precise timing, and recent evidence suggest that timing problems observed in other neurological conditions such as Parkinson's disease, essential tremor, and dystonia may reflect disrupted interactions between the basal ganglia and cerebellum.The complex concepts emerging from this consensus paper should provide a foundation for further discussion, helping identify basic research questions required to understand how the brain represents and utilizes time, as well as delineating ways in which this knowledge can help improve the lives of those with neurological conditions that disrupt this most elemental sense. The panel of experts agrees that timing control in the brain is a complex concept in whom cerebellar circuitry is deeply involved. The concept of a timing machine has now expanded to clinical disorders.


Mathematical model of the changes in heart rate elicited by vagal stimulation.
We developed a mathematical model of the underlying cellular mechanisms responsible for the changes in sinus cycle length (SCL) elicited by vagal stimulation in intact animals. The model incorporated a stimulation-mediated depletion of the releasable pool of acetylcholine (ACh) in the nerve endings, the in vitro reaction kinetics of acetylcholinesterase, and the electrical activity of a pacemaker cell with six membrane ionic currents. SCL increased linearly with the frequency of simulated vagal stimulation, as it does in animal experiments, because the concentration of ACh in the neuroeffector junction [( ACh]) saturated as the frequency of stimulation was increased and because SCL increased geometrically in response to increases in [ACh]. The dependence of SCL on the timing of vagal stimulation in the cardiac cycle resulted, in part, from the dependence of [ACh] on SCL. Simulated vagal stimulation entrained the sinus node because the rate of activation and inactivation of ACh-activated K+ channels depended only weakly on membrane potential during diastolic depolarization. SCL increased geometrically with [ACh], because 1) during diastolic depolarization, the amplitude of the ACh-activated K+ current was approximately equal to the amplitude of the sum of the other ionic currents, 2) [ACh] was low enough to saturate neither acetylcholinesterase nor the cellular system that activates the ACh-activated K+ channels, 3) the pacemaker cell membrane behaved electrotonically like a capacitor, and 4) the sum of all the ionic currents increased linearly with the amplitude of the ACh-activated K+ current.


The Dual Inhibition of RNA Pol I Transcription and PIM Kinase as a New Therapeutic Approach to Treat Advanced Prostate Cancer.
The MYC oncogene is frequently overexpressed in prostate cancer. Upregulation of ribosome biogenesis and function is characteristic of MYC-driven tumors. In addition, PIM kinases activate MYC signaling and mRNA translation in prostate cancer and cooperate with MYC to accelerate tumorigenesis. Here, we investigate the efficacy of a single and dual approach targeting ribosome biogenesis and function to treat prostate cancer. The inhibition of ribosomal RNA (rRNA) synthesis with CX-5461, a potent, selective, and orally bioavailable inhibitor of RNA polymerase I (Pol I) transcription, has been successfully exploited therapeutically but only in models of hematologic malignancy. CX-5461 and CX-6258, a pan-PIM kinase inhibitor, were tested alone and in combination in prostate cancer cell lines, in Hi-MYC- and PTEN-deficient mouse models and in patient-derived xenografts (PDX) of metastatic tissue obtained from a patient with castration-resistant prostate cancer. CX-5461 inhibited anchorage-independent growth and induced cell-cycle arrest in prostate cancer cell lines at nanomolar concentrations. Oral administration of 50 mg/kg CX-5461 induced TP53 expression and activity and reduced proliferation (MKI67) and invasion (loss of ductal actin) in Hi-MYC tumors, but not in PTEN-null (low MYC) tumors. While 100 mg/kg CX-6258 showed limited effect alone, its combination with CX-5461 further suppressed proliferation and dramatically reduced large invasive lesions in both models. This rational combination strategy significantly inhibited proliferation and induced cell death in PDX of prostate cancer. Our results demonstrate preclinical efficacy of targeting the ribosome at multiple levels and provide a new approach for the treatment of prostate cancer. Clin Cancer Res; 22(22); 5539-52. ©2016 AACR.