12 TIPS for Implementing Peer Instruction in Medical Education.
This article was migrated. The article was marked as recommended. Peer Instruction (PI) is a vibrant instructional strategy, used successfully for over two decades in undergraduate physics and mathematics courses. It has had limited use and few publications in medical education. This 12 TIPS provides a focused review on the evidence supporting its use in higher education and rationale for its wider adoption in medical education. The authors detail important steps for its implementation with large classes. Based on several years of experience with PI in a US allopathic medical school, they feel that PI attends to core principles from the science of learning and provides students and faculty with immediate feedback on learning. It is also adaptable to on-line synchronous administration.


Effects of prostanoids on isolated feline cerebral arteries. II. Roles of extra- and intracellular calcium for the prostaglandin F2 alpha-induced contraction.
The roles of extra- and intracellular calcium for the contractile effects of PGF2 alpha in the feline basilar artery (BA) were investigated. Comparisons were made with contractions induced by K+ and noradrenaline (NA). Addition of nifedipine to PGF2 alpha- or K+ (124 mM)-contracted arteries resulted in an incomplete relaxation, whereas NA-contracted vessels were completely relaxed. Incubation of the preparations in a calcium-free medium containing 10(-5) M EGTA for 5-10 min almost abolished contractions induced by K+ and NA. In contrast, 63% of the response to PGF2 alpha remained after pretreatment of the arteries in a calcium-free solution for 40 min; PGF2 alpha produced a biphasic contraction in 17 out of 20 preparations consisting of a rapidly developing initial phase followed by a second increase in tension after 1-6 min. The second phase was absent if the EGTA-concentration was increased to 10(-4) M, or if the arteries were pre-treated with nifedipine. After incubation of the arteries in a calcium-free medium for 40-120 min and K+-depolarization, re-addition of calcium elicited contractions at lower concentrations in the presence of PGF2 alpha than in controls. The results suggest that PGF2 alpha-induced contractions in the feline BA are considerably less dependent on extracellular calcium than contractions evoked by K+ or NA. PGF2 alpha appears to be able to release calcium from two cellular stores, and may also promote calcium influx through the cell membrane.


[Possible mechanisms of the emergence of chromosome aberrations. VIII. Cytogenetic analysis of the dynamics of repair and occurrence of potential chromosome damage in bone marrow cells of gamma-irradiated mice].
The dynamics of mitosis delay and appearance of structural chromosomal aberrations (SCA) in relation to radiation dose and time after postradiation are studied. The results obtained suggest that potential DNA damage (PD)--chemical modifications responsible for SCA formation--induce a G2-block. According to this result, it was concluded that PD arose in 30, 80, 90, and 95% of cells at radiation doses of 9, 24, 51, and 75 cGy, respectively. In cycling cells, PD are either repaired or become SCA. If within the first hour after radiation, it was more probable that they would be repaired. The rate of PD repair decreased and the probability of SCA increased as the length of time after radiation increased. The probability of PD also depended on the separation rate of sister chromatids in mitosis. Radiation affects the rate of chromatid separation and, consequently, the occurrence of PD.


Direct long-term effect of hydrocortisone on insulin and glucagon release from mouse pancreatic islets in tissue culture.
The effects of glucocorticoids on the pancreatic endocrine function was studied in isolated mouse pancreatic islets maintained in tissue culture for 1 to 3 weeks. Following culture for 2 week without corticoid supplement acute experiments with hydrocortisone showed no significant effect on the glucose-induced insulin release at 10(-8) to 10(-5) mol/l hydrocortisone. When, however, the islets were cultured in the presence of hydrocortisone, there was an increased insulin release to the medium in a dose-dependent manner, with the maximal effect at 10(-7) mol/l hydrocortisone. The release of glucagon to the medium was not affected to the same degree, but showed a slight inhibition at increasing concentrations of hydrocortisone. Short-term experiments after the culture period showed that islets cultured for 3 weeks in the presence of 10(-7) to 10(-5) mol/l hydrocortisone had an enhanced insulin secretion in response to glucose. The islets did not show any statistically significant change in their insulin- and DNA-content after 3 weeks of culture with hydrocortisone, but a marked reduction in the content of hydrocortisone. The present results suggest that physiological concentrations of hydrocortisone are importance for mouse islets to maintain their insulin production in tissue culture.


Tumors resurrect an embryonic vascular program to escape immunity.
Tumors can escape immunity through multiple mechanisms, one of which is by enforcing a state of unresponsiveness of the tumor vasculature to inflammatory cytokines. This results in a lack of adhesiveness of angiogenic endothelial cells for immune cells and thus compromised immunity. This type of escape from immunity, called tumor endothelial cell anergy, is the result of exposure to angiogenic growth factors. Angiogenesis is a hallmark not only of cancer but also of embryonic development. It is assumed that angiogenesis-induced suppression of adhesion molecules is a regulatory function to provide an embryo with immune privileged conditions and allow uninterrupted growth and development. It is becoming clear that similar conditions are used by tumors to evade the immune system and ensure progressive growth. Gaining enhanced insight into these immune-privileged conditions is important as endothelial cell anergy can be overcome by angiogenesis inhibitors, an application that is rapidly emerging as a successful strategy to improve immunotherapy. The literature on endothelial adhesion molecule expression and leukocyte-vessel wall interactions during embryonic and fetal development is sparse, but available data allow the hypothesis that tumors, through angiogenesis, enforce an embryonic-like gene expression program in endothelial cells to suppress leukocyte infiltration and compromise antitumor immunity.


Oral transmucosal fentanyl citrate (OTFC) for the treatment of breakthrough pain in cancer patients: a controlled dose titration study.
Oral transmucosal fentanyl citrate (OTFC) is a novel opioid formulation in which the potent synthetic mu-agonist fentanyl is embedded in a sweetened matrix that is dissolved in the mouth. It is undergoing investigation as a treatment for cancer-related breakthrough pain, a prevalent phenomenon defined as a transitory flare of moderate to severe pain that interrupts otherwise controlled persistent pain. There have been no controlled trials of other treatments for this condition. To evaluate the safety and efficacy of ascending doses of OTFC, a novel controlled dose titration methodology was developed that applied blinding and randomization procedures to the evaluation of recurrent pains in the home environment. The study was a multicenter, randomized, double-blind dose titration study in ambulatory cancer patients. The sample comprised adult patients receiving a scheduled oral opioid regimen equivalent to 60-1000 mg oral morphine per day, who were experiencing at least one episode per day of breakthrough pain and had achieved at least partial relief of this pain by use of an oral opioid rescue dose. After collection of 2 days of baseline data concerning the efficacy of the usual rescue drug, patients were randomly treated with either 200 or 400 microg OTFC unit doses in double-blind fashion. Up to two breakthrough pains each day could be treated with up to four OTFC unit doses per pain. OTFC in unit doses containing 200, 400, 600, 800, 1200 or 1600 microg of fentanyl citrate were available for the study. The unit dose was titrated upward in steps until the patient had 2 consecutive days on which breakthrough pain could be treated with the single unit dose, titration was ineffective at a 1600 microg unit dose, or 20 days elapsed. To maintain the double-blind, orders to titrate up were ignored one-third of the time according to a pre-defined randomization schedule accessible only to an unblinded study pharmacist. Main outcome measures included, numeric or categorical measures of pain intensity, pain relief, and global assessment of drug performance. Dose response relationships were found suggesting that the methodology was sensitive to opioid effects. Seventy-four percent of patients were successfully titrated. There was no relationship between the total daily dose of the fixed schedule opioid regimen and the dose of OTFC required to manage the breakthrough pain. Although the study was not designed to provide a definitive comparison between OTFC and the usual rescue drug, exploratory analyses found that OTFC provided significantly greater analgesic effect at 15, 30 and 60 min, and a more rapid onset of effect, than the usual rescue drug. Adverse effects of the OTFC were typically opioid-related, specifically somnolence, nausea and dizziness. Very few adverse events were severe or serious. This study demonstrated the feasibility of controlled trial methodology in studies of breakthrough pain. OTFC appears to be a safe and effective therapy for breakthrough pain, and dose titration can usually identify a unit dose capable of providing adequate analgesia. If the lack of a relationship between the effective OTFC dose and fixed schedule opioid regimen is confirmed, dose titration may be needed in the clinical use of this formulation. Further investigation of OTFC as a specific treatment for breakthrough pain is warranted.


Three-dimensional tissue models for drug discovery and toxicology.
Launching a new drug on the market is an extremely time-consuming and expensive process. The total costs from the lab bench to the patient's bedside are in the range of $800 million for each new compound. Innovative pre-clinical assays are urgently needed to select the most promising drug candidates. High-throughput molecular screening does not provide information on the effects on cellular functions. Testing on animals is expensive, ethically controversial, and poorly predictive of the response in humans. Conventional two-dimensional (2D) cellular assays do not accurately reflect the drug response in vivo. To overcome these limitations, biotechnologists are developing three-dimensional (3D) cultures. 3D cultures provide more accurate compound screening and can eliminate toxic and ineffective substances at an early stage. Moreover, 3D cultures can accomplish the 3R agenda (refinement, reduction, and replacement) for the replacement of toxicity testing on animals. We provide an up-to-date overview on the patents in the field.


[Mechanisms of the interaction of the leukemic cell with endogenous regulators of the proliferation of hematopoietic tissue].
A substance inhibiting DNA synthesis in mouse leukemic cells was isolated from the regenerating calf spleen. When added to a suspension of leukaemic cells, this substance is adsorbed on their surface. The following changes in cell features being noticed: 1) a minute decrease in electrophoretic cell motility, 2) a decrease in esterase activity of the cells, 3) an increase in microviscosity of membrane lipids, 4) an increase in the intracellular pH values. With a longer contact with this substance, changes in nuclear chromatin structure were noticed, with special reference to weakened bonds between DNA and proteins. The data obtained are of significance for revealing molecular mechanisms of hematopoietic mediator action on target cells.


Anesthesia for trans-sternal thymectomy: modified non-muscle relaxant technique.
Anesthesia for thymectomy in myasthenia gravis is challenging. Early surgical management is now considered to be an important therapeutic intervention for most of the patients of myasthenia gravis. The anesthetic experience of that technique is quite large. It involves either muscle relaxant or non-muscle relaxant techniques. However, the literature is deficient of standard anesthetic technique for thymectomy. Therefore we present in this report a modified non-muscle relaxant technique for thymectomy. We report one case with thymectomy under general anesthesia using fentanyl and propofol for induction and endotracheal intubation using non-muscle relaxant technique. The intubating, intraoperative and postoperative conditions were excellent.


LacI(Ts)-regulated expression as an in situ intracellular biomolecular thermometer.
In response to needs for in situ thermometry, a temperature-sensitive vector was adapted to report changes in the intracellular heat content of Escherichia coli in near-real time. This model system utilized vectors expressing increasing quantities of β-galactosidase in response to stepwise temperature increases through a biologically relevant range (22 to 45°C). As judged by calibrated fluorometric and colorimetric reporters, both whole E. coli cells and lysates expressed significant repeatable changes in β-galactosidase activity that were sensitive to temperature changes of less than 1°C (35 to 45°C). This model system suggests that changes in cellular heat content can be detected independently of the medium in which cells are maintained, a feature of particular importance where the medium is heterogeneous or nonaqueous, or otherwise has a low heat transfer capacity. We report here that the intracellular temperature can be reliably obtained in near-real time using reliable fluorescent reporting systems from cellular scales, with a 20°C range of detection and at least 0.7°C sensitivity between 35 and 45°C.


Factors associated with the absence of pharmacological treatment for common modifiable complications in children with chronic kidney disease.
Chronic kidney disease (CKD) is associated with many comorbidities requiring complex management. We described treatment patterns for common modifiable CKD complications (high blood pressure, anemia, hyperphosphatemia, and acidosis) according to severity of CKD and examined factors associated with the absence of drug therapy, among participants with a persistent comorbidity, for 1 year in children enrolled in the CKiD study. A total of 703 CKiD participants contributed 2849 person-visits over a median 3.5 years of follow-up. Using pairs of annual visits, we examined whether participants with abnormal biomarker levels at the first (index) visit persisted in the abnormal levels 1 year later according to CKD risk stage. Multivariate analyses identified demographic and clinical factors associated with the absence of drug therapy among those with persistent comorbid conditions for 1 year. The overall proportions of person-visits prescribing therapy at 1-year follow-up for treating anemia, acidosis, hyperphosphatemia, and high blood pressure were 54%, 45%, 29%, and 81%, respectively. The frequency of therapy increased with advanced CKD risk stage for all comorbidities; however, 19-23% of participants with acidosis, 24-27% with anemia, and 30-39% with hyperphosphatemia at high-risk stages (E and F) were not prescribed appropriate therapy despite the persistence of abnormal levels of these biomarkers for at least 1 year. The resolution of comorbidities at advanced CKD stages without treatment was unlikely. Many children with CKD in the CKiD cohort did not receive pharmacological treatment for common and persistent modifiable comorbidities, even in severe CKD risk stages.


Relation between cardiovascular and metabolic disease and cognition in very old age: cross-sectional and longitudinal findings from the berlin aging study.
This study documented findings on the relation between cognitive functioning (perceptual speed, memory, fluency, and knowledge) and cardiovascular and metabolic disease in a sample of very old adults (ages 70 and older), both cross-sectionally (n=516) and longitudinally (n=206) in a 4-year follow-up. After age, SES, sex, and dementia status were controlled for, 4 diagnoses were negatively associated with cognition: congestive heart failure, stroke, coronary heart disease, and diabetes mellitus, with a joint effect of 0.47 standard deviations. The impact of disease status was largest on perceptual speed and fluency, memory was impacted only by diabetes, and knowledge was not related to any somatic diagnosis. There was no differential decline in participants diagnosed with 1 of these 4 diseases and those who were not. The only cardiovascular risk factor associated with cognitive performance was alcohol consumption.


BI-1 regulates endoplasmic reticulum Ca2+ homeostasis downstream of Bcl-2 family proteins.
BI-1 (Bax inhibitor-1) is an evolutionarily conserved multitransmembrane protein that resides in the endoplasmic reticulum (ER) and that has documented cytoprotective functions in both animals and plants. Recent studies indicate that BI-1 shares in common with Bcl-2/Bax family proteins the ability to regulate the amounts of Ca(2+) that can be released from the ER by agents, such as the ER-Ca(2+)-ATPase (SERCA) inhibitor thapsigargin (TG). Using an ER-targeted, Ca(2+) indicator (cameleon), with characteristics optimized for measuring ER Ca(2+) ([Ca(2+)](er)), we studied the effects of BI-1 on [Ca(2+)](er) in resting and TG-treated cells. Similar to cells overexpressing antiapoptotic Bcl-2 or Bcl-X(L), overexpression of BI-1 resulted in lower resting [Ca(2+)](er), with concomitantly less Ca(2+) released into the cytosol upon stimulation by TG and with a higher rate of Ca(2+) leakage from the ER. Co-expression of SERCA restored levels of [Ca(2+)](er) to normal, showing opposing actions of the ER-Ca(2+)ATPase and BI-1 on ER Ca(2+) homeostasis. Conversely, cells with deficient BI-1 have increased [Ca(2+)](er), and release more Ca(2+) into the cytosol when challenged with TG. In BI-1-deficient cells, Bcl-X(L) fails to reduce [Ca(2+)](er), indicating that BI-1 functions downstream of Bcl-X(L). In bax(-/-)bak(-/-) double knock-out cells, both BI-1 and Bcl-X(L) retained their ability to reduce [Ca(2+)](er), suggesting that BI-1 and Bcl-X(L) operate downstream of or parallel to Bax/Bak. The findings reveal a hierarchy of functional interactions of BI-1 with Bcl-2/Bax family proteins in regulating ER Ca(2+) homeostasis.


Psychometric evaluation of the Health-Risk Attitude Scale (HRAS-13): assessing the reliability, dimensionality and validity in the general population and a patient population.
The aim of this study was to assess the reliability, dimensionality and validity of the self-report questionnaire Health-Risk Attitude Scale (HRAS-13) in a sample of the general population and a patient population. Sample 1 (n = 930) was recruited from the general population aged 18-65 years in the Netherlands. Sample 2 (n = 486) was recruited from the population of knee and hip osteoarthritis patients aged 45 and over, also from the Netherlands. Reliability was assessed using Cronbach's alpha, average inter-item correlation and item-total correlations. Dimensionality was examined using confirmatory factor analysis (CFA), principal component analysis (PCA) and bifactor analysis. Validity was assessed by performing known-group analysis using ANOVA tests. Cronbach's alphas of the HRAS-13 were 0.73 in sample 1 and 0.69 in sample 2. Reliability and dimensionality analyses differed slightly between the samples, and suggest that a short version of the HRAS may capture a general component of health-risk attitude. Validity assessment of known groups showed that the HRAS-13 and a likely HRAS-6 distinguished between subgroups of respondents based on most of the assessed characteristics, but not all. These findings are a preliminary indication that the HRAS-13 is a promising multidimensional instrument for measuring health-risk attitude. However, further research in various samples on decisions where health risks play a role is warranted to confirm the dimensionality of the HRAS-13 and the items to be retained in a full or a shorter version.


Realigning biomedical science instruction in predoctoral curricula: a proposal for change.
Currently in North America, there is an active dialogue going on about the state of predoctoral dental education and the need for curriculum change, innovation, and the adoption of contemporary, competency-based educational models. At the institutional level, curriculum committees struggle with requests from faculty to add new content to an overburdened didactic and clinic schedule. This article will describe potential solutions centering on the role and scope of the biomedical sciences in predoctoral dental education. The authors propose that dental educators and institutions reconsider the current admission prerequisites and curriculum content of the biomedical sciences in predoctoral programs. The proposed changes are intended to eliminate content redundancy between undergraduate and predoctoral dental education by integration of the biomedical sciences--in particular, biochemistry, microbiology, and physiology--into other clinically oriented coursework and learning experiences in the curriculum based on a pathophysiology model that fosters students' comprehension of the etiology of oral and systemic diseases encountered by the general dental practitioner. The authors explore how changes in the biomedical science prerequisites for dental school matriculation and associated modifications in curriculum focus and content would impact admissions testing, composition of national board exams, and strategies for teaching and learning within dental schools.


Perspective on 'Harm' in Personalized Medicine.
Avoiding harm is an uncontroversial aim of personalized medicine and other epidemiologic initiatives. However, the precise mathematical translation of "harm" is disputable. Here we use a formal causal language to study common, but distinct, definitions of "harm". We clarify that commitment to a definition of harm has important practical and philosophical implications for decision making. We relate our practical and philosophical considerations to ideas from medical ethics and legal practice.


Body weight gain rate in patients with Parkinson's disease and deep brain stimulation.
We evaluated body weight changes in patients with Parkinson's disease (PD) after electrode implantation for deep brain stimulation (DBS) in the subthalamic nucleus (STN) in relation to clinical improvement. Thirty PD patients who received STN DBS were included (22 men, 8 women; mean age, 60.0 +/- 7.1 years; mean PD duration, 13.5 +/- 3.7 years; mean body mass index [BMI], 21.6 +/- 3.0 kg/m2). Body weight, physical activity, and Unified Parkinson's Disease Rating Scale (UPDRS) scores were noted before and 3 and 12 months after the procedure. Significant weight gain occurred in 29 patients; the mean increase was 14.8 +/- 9.8% of initial body weight in 1 year. Of the patients, 46.5% reported weight gain in the first 3 months, 21.4% gradual weight gain in the first 6 months, and 32.1% a slow increase for 1 year. Mean BMI increased up to 24.7 +/- 3.7 kg/m2. After 1 year, mean UPDRS motor score improved significantly in off and in on; and therapy complications improved by 91.0 +/- 17.0%. BMI changes at 3 and 12 months were significantly correlated to dyskinesia score changes, and levodopa dosage was not. In PD, STN DBS produces not only symptom control, but also weight gain. DBS candidates should be given nutritional counseling before the intervention to prevent rapid and/or excessive weight gain.


Quantitative gene expression profiles in real time from expressed sequence tag databases.
An accumulation of expressed sequence tag (EST) data in the public domain and the availability of bioinformatic programs have made EST gene expression profiling a common practice. However, the utility and validity of using EST databases (e.g., dbEST) has been criticized, particularly for quantitative assessment of gene expression. Problems with EST sequencing errors, library construction, EST annotation, and multiple paralogs make generation of specific and sensitive qualitative arid quantitative expression profiles a concern. In addition, most EST-derived expression data exists in previously assembled databases. The Virtual Northern Blot (VNB) (http: //tlab.bu.edu/vnb.html) allows generation, evaluation, and optimization of expression profiles in real time, which is especially important for alternatively spliced, novel, or poorly characterized genes. Representative gene families with variable nucleotide sequence identity, tissue specificity, and levels of expression (bcl-xl, aldoA, and cyp2d9) are used to assess the quality of VNB's output. The profiles generated by VNB are more sensitive and specific than those constructed with ESTs listed in preindexed databases at UCSC and NCBI. Moreover, quantitative expression profiles produced by VNB are comparable to quantization obtained from Northern blots and qPCR. The VNB pipeline generates real-time gene expression profiles for single-gene queries that are both qualitatively and quantitatively reliable.


Cationic Defect-Modulated Li-Ion Migration in High-Voltage Li-Metal Batteries.
Li metal exhibits high potential as an anode material for next-generation high-energy density batteries. However, the nonuniform transport of Li+ ions causes Li-dendrite growth at the metal electrode, leading to severe capacity decay and a short cycling life. In this study, negatively charged lithiophilic sites (such as cationic metal vacancies) were used as hosts to regulate the atomic-scale Li+-ion deposition in Li-metal batteries (LMBs). As a proof of concept, three-dimensional (3D) carbon nanofibers (CNFs) decorated with negatively charged TiNbO4 grains (labeled CNF/nc-TNO) were confirmed to be promising Li hosts. Cationic vacancies caused by the carbothermal reduction of Nb5+ and Ti4+ ions generated a negatively charged fiber surface and strong electrostatic interactions that guided the Li+-ion flux to the shadowed areas underneath the fiber and throughout the fibrous mat. Consequently, circumferential Li-metal plating was observed in the CNF/nc-TNO host, even at a high current density of 10 mA cm-2. Moreover, CNF/nc-TNO asymmetric cells delivered a significantly more robust and stable Coulombic efficiency (CE) (99.2% over 380 cycles) than cells comprising electrically neutral CNFs without cationic defects (which exhibits rapid failure after 20 cycles) or Cu foil (which exhibits rapid CE decay, with a CE of 87.1% after 100 cycles). Additionally, CNF/nc-TNO exhibited high stability and low-voltage hysteresis during repeated Li plating/stripping (for over 4000 h at 2 mA cm-2) with an areal capacity of 2 mAh cm-2. It was further paired with high-voltage LiNi0.8Co0.1Mn0.1 (NCM811) cathodes, and the full cells showed long-term cycling (220 cycles) with a CE of 99.2% and a steady rate capability.


GK921, a transglutaminase inhibitor, strengthens the antitumor effect of cisplatin on pancreatic cancer cells by inhibiting epithelial-to-mesenchymal transition.
Pancreatic adenocarcinoma (PAAD), a common digestive malignant tumor, presents high mortality rates and limited treatment methods. Currently, chemotherapy remains the main therapy method for patients with PAAD. As a classical chemotherapy drug, cisplatin (DDP) is limited by dose-related toxicity in patients with PAAD. In this study, we demonstrated that TGM2 may be a treatment and prognosis marker in pancreatic cancer patients. Co-treatment of low dose of DDP and GK921, a transglutaminase (TGM2) inhibitor, is capable of synergistically inhibiting the PAAD cell viability and proliferation in vitro and in vivo. Based on in vitro study, GK921 inhibited the epithelial-to-mesenchymal transition (EMT) induced by TGM2 as well as aggravated cell cycle arrest and apoptosis resulted from DDP, making pancreatic cancer cells more sensible to DDP. Our results showed that GK921 increased the protein levels regarding E-cadherin as well as decreased the protein level regarding Snail2, N-cadherin, which indicated that GK921 inhibited EMT in pancreatic cancer cells. Snail2 overexpression inhibited GK921/DDP-induced cell apoptosis, as well as mitigated the GK921/DDP-caused cell death and the EMT inhibition. In vivo studies also found GK921/DDP combination can further inhibit the growth of PAAD without significantly side effects. To sum up, we showed that GK921 increased PAAD cells sensitivity to DDP via inhibiting EMT. As revealed, DDP/GK921 co-treatment could promisingly serve for treating PAAD patients.


Visual processing asymmetries in change detection.
Change detection is critically dependent on attentional mechanisms. However, the relation between an asymmetrical distribution of visuo-spatial attention and the detection of changes in visual scenes is not clear. Spatial tasks are known to induce a stronger activation of the right hemisphere. The effects of such visual processing asymmetries induced by a spatial task on change detection were investigated. When required to detect changes in the left and in the right visual fields, participants were significantly faster in detecting changes on the left than on the right. Importantly, this left-side superiority in change detection is not influenced by inspection time, suggesting a critical role of visual processing benefit for the left visual field.


Lipidomic Analysis of the Outer Membrane Vesicles from Paired Polymyxin-Susceptible and -Resistant Klebsiella pneumoniae Clinical Isolates.
Gram-negative bacteria produce outer membrane vesicles (OMVs) as delivery vehicles for nefarious bacterial cargo such as virulence factors, which are antibiotic resistance determinants. This study aimed to investigate the impact of polymyxin B treatment on the OMV lipidome from paired polymyxin-susceptible and -resistant Klebsiella pneumoniae isolates. K. pneumoniae ATCC 700721 was employed as a reference strain in addition to two clinical strains, K. pneumoniae FADDI-KP069 and K. pneumoniae BM3. Polymyxin B treatment of the polymyxin-susceptible strains resulted in a marked reduction in the glycerophospholipid, fatty acid, lysoglycerophosphate and sphingolipid content of their OMVs. Conversely, the polymyxin-resistant strains expressed OMVs richer in all of these lipid species, both intrinsically and increasingly under polymyxin treatment. The average diameter of the OMVs derived from the K. pneumoniae ATCC 700721 polymyxin-susceptible isolate, measured by dynamic light scattering measurements, was ~90.6 nm, whereas the average diameter of the OMVs isolated from the paired polymyxin-resistant isolate was ~141 nm. Polymyxin B treatment (2 mg/L) of the K. pneumoniae ATCC 700721 cells resulted in the production of OMVs with a larger average particle size in both the susceptible (average diameter ~124 nm) and resistant (average diameter ~154 nm) strains. In light of the above, we hypothesize that outer membrane remodelling associated with polymyxin resistance in K. pneumoniae may involve fortifying the membrane structure with increased glycerophospholipids, fatty acids, lysoglycerophosphates and sphingolipids. Putatively, these changes serve to make the outer membrane and OMVs more impervious to polymyxin attack.


Correlation of serum 1,25-dihydroxycholecalciferol with the incidence of primary open-angle glaucoma: A cross-sectional study on patients with chronic illnesses.
Primary open-angle glaucoma (POAG) is a clinical progressive neuropathy which can lead to irreversible blindness if left untreated. A low level of serum Vitamin D3 is a major risk factor for glaucoma, and hence, represents a second target for glaucoma therapy following intraocular pressure (IOP). However, there is still controversy about whether there is a direct correlation between Vitamin D3 deficiency and the risk of increased IOP. This study aims to investigate the correlation between low serum levels of 1,24-dihydroxycholecalciferol and the development of open-angle glaucoma. The study included a total of forty-one patients with POAG. Patients were classified into whether they have chronic illnesses such as type 2 diabetes and hypertension. Matching control subjects of 20 healthy controls were also included in the study. Anthropometric measures and venous blood samples were taken from all participants for serum analysis of various biochemical markers including serum 1,25-dihydroxycholecalciferol (1,25(OH)2D) levels. Overall, serum Vitamin D3 levels were 15% significantly lower in the patient's cohort with open-angle glaucoma as compared to the healthy participants (P < 0.05). Among those, 63% of type 2 diabetic participants had significantly low levels of Vitamin D3 (P < 0.01). There was also a significant 70% reduction in serum Vitamin D3 levels among the hypertensive participants, (P < 0.001). We concluded that lower serum 1,25(OH) 2D levels were significantly associated with an increased risk of open-angle glaucoma in patients with chronic illnesses.


Drug Abuse Ontology to Harness Web-Based Data for Substance Use Epidemiology Research: Ontology Development Study.
Web-based resources and social media platforms play an increasingly important role in health-related knowledge and experience sharing. There is a growing interest in the use of these novel data sources for epidemiological surveillance of substance use behaviors and trends. The key aims were to describe the development and application of the drug abuse ontology (DAO) as a framework for analyzing web-based and social media data to inform public health and substance use research in the following areas: determining user knowledge, attitudes, and behaviors related to nonmedical use of buprenorphine and illicitly manufactured opioids through the analysis of web forum data Prescription Drug Abuse Online Surveillance; analyzing patterns and trends of cannabis product use in the context of evolving cannabis legalization policies in the United States through analysis of Twitter and web forum data (eDrugTrends); assessing trends in the availability of novel synthetic opioids through the analysis of cryptomarket data (eDarkTrends); and analyzing COVID-19 pandemic trends in social media data related to 13 states in the United States as per Mental Health America reports. The domain and scope of the DAO were defined using competency questions from popular ontology methodology (101 ontology development). The 101 method includes determining the domain and scope of ontology, reusing existing knowledge, enumerating important terms in ontology, defining the classes, their properties and creating instances of the classes. The quality of the ontology was evaluated using a set of tools and best practices recognized by the semantic web community and the artificial intelligence community that engage in natural language processing. The current version of the DAO comprises 315 classes, 31 relationships, and 814 instances among the classes. The ontology is flexible and can easily accommodate new concepts. The integration of the ontology with machine learning algorithms dramatically decreased the false alarm rate by adding external knowledge to the machine learning process. The ontology is recurrently updated to capture evolving concepts in different contexts and applied to analyze data related to social media and dark web marketplaces. The DAO provides a powerful framework and a useful resource that can be expanded and adapted to a wide range of substance use and mental health domains to help advance big data analytics of web-based data for substance use epidemiology research.


Supraspinous ligament avulsion: a case report.
Avulsion of a supraspinous ligament is one potential cause of low back pain. Since the symptom pattern of this condition may mimic that of other, more common, conditions, a thorough biomechanical and radiological examination is necessary to ensure proper and successful management. The necessity for a thorough and complete biomechanical, laboratory and x-ray evaluation is emphasized before successful treatment can be initiated.


Lipoamino acids as major components of absorption promoters in drug delivery.
Many biologically active compounds are unsuitable for development as drugs due to their poor bioavailability. For hydrophilic compounds, modifications to increase lipophilicity can increase passive diffusion or increase uptake into the lymphatic system. Alternatively, improved bioavailability of hydrophilic drug candidates may be achieved by formulation with absorption promoters such as surfactants, penetration enhancers, or ion pairing agents. This approach to enhancing bioavailability also has the potential to widen the range of compound categories that can be used as chemical probes to study biological systems in cells and in vivo where membrane permeability would otherwise be a significant limitation. Lipidic amino acids, which combine the structural properties of lipids with those of α-amino acids, represent a relatively unexplored class of agents that can improve drug adsorption. This review discusses the potential of absorption promoters possessing lipoamino acids for improving drug bioavailability.


Genetic parameters for pathogen-specific mastitis resistance in Danish Holstein Cattle.
The objective of this study was to estimate heritabilities for and genetic correlations among different pathogen-specific mastitis traits. The traits were unspecific mastitis, which is all mastitis treatments regardless of the causative pathogen as well as mastitis caused by Streptococcus dysgalactiae, Escherichia coli, coagulase-negative staphylococci (CNS), Staphylococcus aureus and Streptococcus uberis. Also groups of pathogens were investigated, Gram-negative v. Gram-positive and contagious v. environmental pathogens. Data from 168 158 Danish Holstein cows calving first time between 1998 and 2006 were used in the analyses. Variances and covariances were estimated using uni- and bivariate threshold models via Gibbs sampling. Posterior means of heritabilities of pathogen-specific mastitis were lower than the heritability of unspecific mastitis, ranging from 0.035 to 0.076 for S. aureus and S. uberis, respectively. The heritabilities of groups of pathogen ranged from 0.053 to 0.087. Genetic correlations among the pathogen-specific mastitis traits ranged from 0.45 to 0.77. These estimates tended to be lowest for bacteria eliciting very different immune responses, which can be considered as the overall pleiotropic effect of genes affecting resistance to a specific pathogen, and highest for bacteria sharing characteristics regarding immune response. The genetic correlations between the groups of pathogens were high, 0.73 and 0.83. Results showed that the pathogen-specific traits used in this study should be considered as different traits. Genetic evaluation for pathogen-specific mastitis resistance may be beneficial despite lower heritabilities than unspecific mastitis because a pathogen-specific mastitis trait is a direct measure of an udder infection, and because the cost of a mastitis case caused by different pathogens has been shown to differ greatly. Sampling bias may be present because there were not pathogen information on all mastitis treatments and because some farms do not record pathogen information. Therefore, improved recording of pathogen information and mastitis treatments in general is critical for a successful genetic evaluation of udder health. Also, economic values have to be specified for each pathogen-specific trait separately.


Cognitive deficits in obstructive sleep apnea: Insights from a meta-review and comparison with deficits observed in COPD, insomnia, and sleep deprivation.
Obstructive sleep apnea (OSA) is a nocturnal breathing disorder that is associated with cognitive impairment. The primary determinants of cognitive deficits in OSA are thought to be sleep disruption and blood gas abnormalities. Cognitive impairment is also seen in other disorders that are characterised primarily by sleep disturbance (e.g., sleep restriction/deprivation, insomnia) or hypoxia/hypercarbia (e.g., chronic obstructive pulmonary disease (COPD)). Assessment of the cognitive deficits observed in these other disorders could help better define the mechanisms underlying cognitive deficits in OSA. This study utilised meta-review methodology to examine the findings from systematic reviews and meta-analyses of the effects of untreated OSA, COPD, insomnia, and sleep deprivation on cognitive function in adults, compared with norms or controls. Eighteen papers met inclusion criteria: seven in OSA, two in insomnia, five in COPD, and four in sleep deprivation. OSA and COPD were both accompanied by deficits in attention, memory, executive function, psychomotor function, and language abilities, suggesting that hypoxia/hypercarbia may be an important determinant of deficits in these domains in OSA. Both OSA and sleep deprivation studies were accompanied by deficits in attention and memory, suggesting that short-term sleep disturbance in OSA may contribute to deficits in these domains. Visuospatial deficits were unique to OSA, suggesting the contribution of a mechanism other than sleep disturbance and hypoxia/hypercarbia to this problem. Our findings suggest that the cognitive deficits associated with untreated OSA are multidimensional, with different physiological disturbances responsible for differing cognitive problems.


Update on low-dose corticosteroids.
Low-dose hydrocortisone is recommended in patients with septic shock unresponsive to fluid and vasopressor therapy. Recent research added new data for patients with septic shock and other target groups such as patients with severe sepsis, acute respiratory distress syndrome (ARDS), community-acquired pneumonia, and burns. The objective of this review is to summarize and comment recent findings on low-dose corticosteroids (LDC) in critically ill patients. In the last 2 years, a series of clinical trials and retrospective analyses investigated LDC therapy in critically ill patients with severe systemic inflammation of various origins. Improvement in morbidity has been demonstrated in ARDS and community-acquired pneumonia. Retrospective propensity-score analyses also suggest that LDC administered in severe septic shock or in septic shock due to community-acquired pneumonia or intestinal perforation may improve survival. Low-dose hydrocortisone or a corresponding low-dose corticosteroid therapy may improve morbidity in specific target groups of critically ill patients. Beneficial effects on mortality remain to be demonstrated in large-scale randomized controlled trials.


Agenda-setting revisited: When and how do primary-care physicians solicit patients' additional concerns?
Soliciting patients' complete agendas of concerns (aka. 'agenda setting') can improve patients' health outcomes and satisfaction, and physicians' time management. We assess the distribution, content, and effectiveness of physicians' post-chief-complaint, agenda-setting questions. We coded videotapes/transcripts of 407 primary-, acute-care visits between adults and 85 general-practice physicians operating in 46 community-based clinics in two states representing urban and rural care. Measures are the incidence of physicians' questions, their linguistic format, position within visits, likelihood of being responded to, and the nature of such responses. Physicians' questions designed to solicit concerns additional to chief concerns occurred in only 32% of visits (p<.001). Compared to questions whose communication format explicitly solicited 'questions' (e.g., "Do you have any questions?"), those that were formatted so as to allow for 'concerns' (e.g., "Any other concerns?") were significantly more likely to generate some type of agenda item (Chi(2) (1, N=131)=11.96, p=.001), and to do so more frequently when positioned 'early' vs. 'late' during visits (Chi(2) (1, N=73)=4.99, p=.025). Agenda setting is comparatively infrequent. The communication format and position of physicians' questions affects patients' provision of additional concerns/questions. Physicians should increase use of optimized forms of agenda setting.


Exposure to antenatal corticosteroids and reduced respiratory sinus arrhythmia in adult survivors of extremely low birth weight.
Purpose/aim: Antenatal corticosteroid (ACS) therapy has dramatically increased survival rates among extremely low birth weight (ELBW) infants. However, the long-term effects of ACS on autonomic nervous system function have not been explored. Using the world's oldest longitudinally followed cohort of ELBW infants we compared respiratory sinus arrhythmia (RSA) among ELBW survivors whose mothers received ACS (ELBW-S), those who did not (ELBW-NS) and normal birth weight (NBW) controls in their 20 and 30 s. Methods: Resting electrocardiogram (ECG) was recorded from ELBW-S (n = 28), ELBW-NS (n = 36), and matched NBW controls (n = 79) at 22-26 and 29-36 years. Resting RSA was compared across groups via analyses of covariance (ANCOVA), adjusting for sex, medication use, postnatal steroid exposure and the presence of chronic health conditions. RSA was also compared across assessments for each group. Results: At 29-36 years, resting RSA in ELBW-S was significantly lower than in NBW controls. RSA in the ELBW-NS group was intermediate between ELBW-S and NBW groups. Although the ELBW-S group also showed nominally reduced RSA compared to NBW controls at the 22-26-year visit, this difference was not statistically significant. Conclusions: ELBW survivors exposed to ACS had lower RSA than NBW controls during their 30 s, suggestive of a decline in parasympathetic input to heart. ELBW survivors who received ACS may be particularly vulnerable to cardiovascular problems in later life.


Liquid acoustic lens for photoacoustic tomography.
Recently, intensive research has been conducted to accelerate the development of photoacoustic (PA) imaging modality for biomedical applications. The use of acoustic lenses to collect ultrasound signals is of great interest. This Letter presents the design and fabrication of a liquid acoustic diverging lens, which can enlarge the acceptance angle of an ultrasound transducer. This lens possesses an inherent advantage of low acoustic impedance and the convenience to be attached to or detached from a commercial flat transducer. Phantom experiments have been carried out to demonstrate the improvement of using such a liquid lens over using a bare transducer for PA tomography.


Thin-film polymer light emitting diodes as integrated excitation sources for microscale capillary electrophoresis.
We report the use of a thin-film polymer light emitting diode as an integrated excitation source for microfabricated capillary electrophoresis. The polyfluorene-based diode has a peak emission wavelength of 488 nm, an active area of 40 microm x 1000 microm and a thickness of similar 2 mm. The simple layer-by-layer deposition procedures used to fabricate the polymer component allow facile integration with planar chip-based systems. To demonstrate the efficacy of the approach, the polyfluorene diode is used as an excitation source for the detection of fluorescent dyes separated on-chip by electrophoresis. Using a conventional confocal detection system the integrated pLED is successfully used to detect fluorescein and 5-carboxyfluorescein at concentrations as low as 10(-6) M with a mass detection limit of 50 femtomoles. The drive voltages required to generate sufficient emission from the polymer diode device are as low as 3.7 V.


[Control mechanisms in a telemarketing call center and workers' complaints of fatigue and exhaustion].
This study describes the control mechanisms used in a telemarketing call center and compares them with workers' complaints previously filed through the respective trade union. Such complaints related to fatigue, mental exhaustion, and powerlessness in the face of a rigid work organization. The work in various divisions of a company with 2,285 employees was approached using ergonomic analysis methods and analysis of the system's management documents. Control mechanisms were classified according to: call time, content, comportment, volume of calls performed, and results. The conclusion was that the company relies on the telemarketers' mental and affective effort to achieve customer interaction and satisfaction, overcoming the difficulties as they arise without either surpassing the scheduled time limits as set by commercial targets or breaching the established controls.


Effects of Chronic Exercise on Endothelial Progenitor Cells and Microparticles in Professional Runners.
The effects of chronic exposure to exercise training on vascular biomarkers have been poorly explored. Our study aimed to compare the amounts of endothelial progenitor cells (EPCs), and endothelial (EMP) and platelet (PMP) microparticles between professional runners and healthy controls. Twenty-five half-marathon runners and 24 age- and gender-matched healthy controls were included in the study. EPCs (CD34+/KDR+, CD133+/KDR+, and CD34+/CD133+), EMP (CD51+) and PMP (CD42+/CD31+) were quantified by flow-cytometry. All blood samples were obtained after 12 h of fasting and the athletes were encouraged to perform their routine exercises on the day before. As compared with controls, the CD34+/KDR+ EPCs (p=0.038) and CD133+/KDR+ EPCs (p=0.018) were increased, whereas CD34+/CD133+ EPCs were not different (p=0.51) in athletes. In addition, there was no difference in MPs levels between the groups. Chronic exposure to exercise in professional runners was associated with higher percentage of EPCs. Taking into account the similar number of MPs in athletes and controls, the study suggests a favorable effect of exercise on these vascular biomarkers.


Counting mitoses by image processing in Feulgen stained breast cancer sections: the influence of resolution.
Counting of mitotic cells has been shown to be of prognostic value in breast cancer in different retrospective studies. Up to now the number of mitoses is assessed mainly manually according to a standardized but strict protocol. Although such a manual procedure is reasonably reproducible, automatic counting of mitotic cells offers the potential for greater objectivity and reproducibility. This paper describes the influence of resolution on automatic recognition by image processing of mitotic cells in Feulgen stained breast cancer sections. Using the image recording, correction and segmentation procedure described in a previous study, five specimens were analyzed: one was used to serve as a training set and four were put aside for later use as independent test set. For each slide, objects from a pre-selected area were recorded at increasing resolution. For each object, contour features and optical density measurements were computed and stored in a data file for statistical analysis. The results showed that increased resolution using a 40x objective lowered the number of misclassified mitoses compared with a 20x objective (overall mean percentage of misclassified mitoses over training and all test specimens: 20x, 24.57; 40x, 7.96). The number of misclassifications of non-mitoses was almost stable per specimen but varied between specimens (19-42%) due to differences among tissues. Given the improvement in classifying mitoses and the possibility to evaluate interactively the measurement result, the described semi-automated mitoses pre-screener of histological sections may be suitable for further testing in a clinical setting.


The effects of infliximab therapy on health-related quality of life in ulcerative colitis patients.
The impact of infliximab induction and maintenance therapy on health-related quality of life (HRQL) was evaluated in patients with ulcerative colitis (UC). In two placebo-controlled, double-blind studies (the Active Ulcerative Colitis Trials 1 and 2 [ACT 1 and 2]), 728 patients were randomized to placebo or infliximab 5 mg/kg or 10 mg/kg. Infusions were administered at weeks 0, 2, 6, and every 8 wk thereafter, up to week 22 (ACT 2) or 46 (ACT 1). Changes in Inflammatory Bowel Disease Questionnaire (IBDQ) and Medical Outcomes Study 36-Item Short Form Health Survey physical and mental component summary (PCS and MCS, respectively) scores were analyzed. Baseline scores for the pooled patient population indicated substantial impairment in HRQL. Improvement at week 8 in the total IBDQ score was significantly greater in the infliximab 5-mg/kg (40, P < 0.001) and 10-mg/kg (36, P < 0.001) groups compared with the placebo group (28). Improvement at week 8 was also significantly greater in the infliximab 5- and 10-mg/kg groups for the PCS (6.8 and 5.9, respectively) and MCS (5.9 and 6.4, respectively) compared with placebo (PCS = 3.7, MCS = 3.0, P < 0.01 for all comparisons). Continued benefit was seen at weeks 30 and 54 with infliximab maintenance therapy (P < 0.001 for all comparisons). Improvement in total IBDQ score correlated significantly (P < 0.001) with improvement in both PCS and MCS scores, and Mayo score. Infliximab therapy substantially improved HRQL in patients with UC. This benefit was sustained through 1 yr with maintenance infliximab therapy.


More Versus Less Blood Pressure Lowering: An Update.
This article summarizes the results of a recent systematic review and meta-analysis of the safety and efficacy of intensive blood pressure (BP) lowering, including an update with the SPRINT (Systolic Blood Pressure Intervention Trial) results. We discuss the consistency of results within this set of trials (eg, ACCORD [Action to Control Cardiovascular Risk in Diabetes] and SPRINT) and the results in the context of other BP-lowering trials, including the recently published HOPE3 (Heart Outcomes Prevention Evaluation-3) study. This study was a narrative review with updated meta-analysis from systematic review of trials comparing more- versus less-intensive BP lowering. With the addition of SPRINT, there are >20 trials comparing more- versus less-intensive treatment with 54,350 participants overall. Over an average of 3.9 years of follow-up, the average systolic BP was 133 mm Hg in the more-intensive treatment arms and 140 mm Hg in the less-intensive treatment arms. More-intensive BP lowering reduced the risk of major vascular events, with benefits seen across a range of groups defined according to baseline systolic BP (120-139 mm Hg, 140-159 mm Hg, or ≥160 mm Hg), age (<70 or >70 years), and main entry criterion (hypertension, diabetes, or renal disease). The evidence accumulated thus far provides clear evidence of the benefits of BP lowering in the 120- to 140-mm Hg range for various high-risk patient groups. Although intensive BP lowering has side effects, these trials indicate that the benefits will predominate for those at high risk of major vascular events.


Physical Education Teacher Education Leaders' Perceptions on a National Curriculum in Physical Education.
Purpose: Given criticism of P-12 physical education and wide variability in instructional quality and subject matter outcomes in the United States (US), a national curriculum has been debated by some scholars as a mechanism for improving the status of the subject matter. Grounded in the systemic reform (SR) model, the purpose of this study was to explore physical education teacher education (PETE) leaders' perceptions regarding the implementation of a national curriculum. Method: In total, 28 individuals participated in in-depth interviews that were inductively/deductively coded and triangulated. Results: Themes indicated that nationalizing the curriculum has the potential to offer explicit educational goals, substantial pedagogical guidelines, and valid assessments. Despite recognizing the potential benefits of national curriculum, however, the majority of participants were opposed because of the inflexibility of such a system in the culturally and geographically diverse school contexts across the US. Conclusions: The concept of national curriculum can be differently interpreted in different countries based on sociocultural, historical, and contextual factors, and its relevance depends primarily on one's perceptions and previous experiences.


Current status of molecular biomarkers in endometrial cancer.
In spite of the high and increasing incidence of endometrial cancer, our current models for prediction of prognosis and even more treatment response are suboptimal, and molecular biomarkers to assist clinical decision making are needed. In this review an overview is given of the currently known as well as promising prognostic and predictive biomarkers in endometrial carcinoma. Key clinical challenges, where use of molecular biomarkers can meet clinical needs, are highlighted. The current status for the presently most promising prognostic and predictive biomarkers in endometrial carcinoma is reviewed. DNA ploidy, p53 status, hormone receptor level, HER2, stathmin, L1 cell adhesion molecule expression and other biomarkers are discussed in relation to the scientific robustness of various essential steps in biomarker development and (current) clinical applicability for individualizing treatment strategies. Tumour heterogeneity and its consequences for biomarker assessment and the importance of developing standardised tests for implementation are discussed. To improve the development and clinical uptake of biomarkers, several strategies are proposed.


The effect of passive heating and face cooling on perceived exertion during exercise in the heat.
Increased body temperature is thought to be an important component of the higher perception of exertion that is a feature of fatigue during exercise in the heat but a causal relationship has yet to be demonstrated. We have investigated the effect of passive heating on the perception of exertion during a standard bout of exercise and also assessed the effect of cooling the head on compensating for the increased body temperature on the feelings of exertion. Ten male subjects performed a 14-min cycling exercise [average power approximately 63% of maximum power output ( W(max))] at an ambient temperature of 35 degrees C at resting rectal temperature [mean (SD): 37.49 (0.27) degrees C; control (CON) trial] on one occasion, and after sitting in a sauna to raise rectal temperature [mean (SD): 38.95(0.13) degrees C; sauna (SAU) trial]. During the exercise, subjects reported their ratings of overall perceived exertion (RPE), perceived exertion of the legs (RPE(legs)) and thermal comfort (TC). A blood sample was collected by the end of the exercise for determination of plasma glucose, lactate and prolactin and haematocrit. RPE values were significantly elevated after passive heating [mean (SE): 14.5 (0.7) units in CON and 17.2 (0.5) units in SAU, at the end of exercise; P<0.001] as were the RPE(legs) ( P<0.01), while ratings of TC were similar in CON and SAU trials. Passive heating increased blood glucose ( P<0.05) but had no effect on lactate at the end of the exercise. Plasma prolactin was markedly elevated as a result of the sauna exposure [mean (SE): 1598 (152) versus 225 (31) mU l(-1) in SAU and CON trials, respectively; P<0.001]. Six of the subjects repeated the two trials but with the face cooled during exercise (trials CON(FAN) and SAU(FAN)) that was achieved by combining face fanning and spraying the face with a mist of cooled water. Face cooling decreased RPE values after sauna to a point that no differences between the two conditions existed. RPE(legs) scores and heart rate, however, remained higher in SAU(FAN) compared with CON(FAN) ( P<0.05). We conclude that hyperthermia is a causative element of the increased perception of exertion during submaximal exercise in the heat and that the effect of increased core temperature on the feelings of exertion is modulated by face cooling.


Differential requirements for cyclase-associated protein (CAP) in actin-dependent processes of Toxoplasma gondii.
Toxoplasma gondii contains a limited subset of actin binding proteins. Here we show that the putative actin regulator cyclase-associated protein (CAP) is present in two different isoforms and its deletion leads to significant defects in some but not all actin dependent processes. We observe defects in cell-cell communication, daughter cell orientation and the juxtanuclear accumulation of actin, but only modest defects in synchronicity of division and no defect in the replication of the apicoplast. 3D electron microscopy reveals that loss of CAP results in a defect in formation of a normal central residual body, but parasites remain connected within the vacuole. This dissociates synchronicity of division and parasite rosetting and reveals that establishment and maintenance of the residual body may be more complex than previously thought. These results highlight the different spatial requirements for F-actin regulation in Toxoplasma which appear to be achieved by partially overlapping functions of actin regulators.


L-ascorbic acid alpha-glucoside formed by regioselective transglucosylation with rat intestinal and rice seed alpha-glucosidases: its improved stability and structure determination.
The definite structure and chemical stability of a new glucoside of L-ascorbic acid (AA) which was enzymatically glucosylated with rat intestinal and rice seed alpha-glucosidases were reported. The stability of this AA derivative in water under aerobic conditions was proved by its remarkable resistance against enhanced oxidative degradation by heat, Cu2+ ion or ascorbate oxidase, and it was found to have no reducing activity toward radicals. These properties were obviously distinguishable from those of AA. This glucoside was effectively hydrolyzed by alpha-glucosidases which possessed the ability to synthesize itself, resulting in the liberation of AA activity. The conjugate was composed of equimoles of AA and glucose. Nuclear magnetic resonance spectra, mass spectra, pH profiles of ultraviolet spectra and pK(a) value of 3.10 supported the coupling of alpha-glucose to the 2-position of AA. From these results, its structure was assigned 2-O-alpha-D-glucopyranosyl-L-ascorbic acid, being distinct from 6-O-alpha-D-glucopyranosyl-L-ascorbic acid formed with Aspergillus niger alpha-glucosidase. These findings indicate that the 2-O-glucoside formed by regioselective transglucosylation withstands oxidative degradation even in aqueous solutions and it can be used as an available active AA source for multicomponent liquid products.


Rates of DNA sequence evolution in experimental populations of Escherichia coli during 20,000 generations.
We examined rates of DNA sequence evolution in 12 populations of Escherichia coli propagated in a glucose minimal medium for 20,000 generations. Previous work saw mutations mediated by mobile elements in these populations, but the extent of other genomic changes was not investigated. Four of the populations evolved defects in DNA repair and became mutators. Some 500 bp was sequenced in each of 36 genes for 50 clones, including 2 ancestral variants, 2 clones from each population at generation 10,000, and 2 from each at generation 20,000. Ten mutations were found in total, all point mutations including mostly synonymous substitutions and nonsynonymous polymorphisms; all 10 were found in mutator populations. We compared the observed sequence evolution to predictions based on different scenarios. The number of synonymous substitutions is lower than predicted from measured mutation rates in E. coli, but the number is higher than rates based on comparing E. coli and Salmonella genomes. Extrapolating to the entire genome, these data predict about 250 synonymous substitutions on average per mutator population, but only about 3 synonymous substitutions per nonmutator population, during 20,000 generations. These data illustrate the challenge of finding sequence variation among bacterial isolates that share such a recent ancestor. However, this limited variation also provides a useful baseline for research aimed at finding the beneficial substitutions in these populations.


Differentiation of schizophrenia using structural MRI with consideration of scanner differences: A real-world multisite study.
Neuroimaging studies have revealed that patients with schizophrenia exhibit reduced gray matter volume in various regions. With these findings, various studies have indicated that structural MRI can be useful for the diagnosis of schizophrenia. However, multisite studies are limited. Here, we evaluated a simple model that could be used to differentiate schizophrenia from control subjects considering MRI scanner differences employing voxel-based morphometry. Subjects were 541 patients with schizophrenia and 1252 healthy volunteers. Among them, 95 patients and 95 controls (Dataset A) were used for the generation of regions of interest (ROI), and the rest (Dataset B) were used to evaluate our method. The two datasets were comprised of different subjects. Three-dimensional T1-weighted MRI scans were taken for all subjects and gray-matter images were extracted. To differentiate schizophrenia, we generated ROI for schizophrenia from Dataset A. Then, we determined volume within the ROI for each subject from Dataset B. Using the extracted volume data, we calculated a differentiation feature considering age, sex, and intracranial volume for each MRI scanner. Receiver-operator curve analyses were performed to evaluate the differentiation feature. The area under the curve ranged from 0.74 to 0.84, with accuracy from 69% to 76%. Receiver-operator curve analysis with all samples revealed an area under the curve of 0.76 and an accuracy of 73%. We moderately successfully differentiated schizophrenia from control using structural MRI from differing scanners from multiple sites. This could be useful for applying neuroimaging techniques to clinical settings for the accurate diagnosis of schizophrenia.


Genotoxic potential of dental bulk-fill resin composites.
To investigate both genotoxicity and hardening of bulk-fill composite materials applied in 4-mm layer thickness and photo-activated for different exposure times. Three flowable bulk-fill materials and one conventional flowable composite were filled in molds (height: 4mm) and irradiated for 20 or 30s. The top (0mm) and bottom (4mm) specimen surface were mechanically scraped, and eluates (0.01g composite in 1.5ml RPMI 1640 cell culture media) prepared for each material, surface level and irradiation time. Genotoxicity was assessed in human leukocytes using both the alkaline comet assay and cytokinesis-blocked micronucleus assay, and Knoop hardness (KHN) was measured at the top and bottom specimen surface (n=8). At both irradiation times, none of the bulk-fill composites significantly affected comet assay parameters used in primary DNA damage assessment or induced significant formation of any of the scored chromatin abnormalities (number of micronuclei, nuclear buds, nucleoplasmic bridges), whether eluates were obtained from the top or bottom surface. Furthermore, no decrease in KHN from the top to the bottom surface of the bulk-fill materials was observed. On the other hand, the conventional composite irradiated for 20s showed at 4-mm depth a significant increase in the percentage of DNA that migrated in the tail and a significant increase in the number of nuclear buds, as well as a significant decrease in KHN relative to the top surface. Bulk-fill resin composites, in contrast to conventional composite, applied in 4-mm thickness and photo-activated for at least 20s do not induce relevant genotoxic effects or mechanical instability.


Phosphorylation, expression and function of the Ultrabithorax protein family in Drosophila melanogaster.
Alternative splicing of the Ultrabithorax homeotic gene transcript generates a family of five proteins (UBX isoforms) that function as transcription factors. All isoforms contain a homeodomain within a common 99 aa C-terminal region (C-constant region) which is jointed to a common 247 aa N-terminal (N-constant) region by different combinations of three small optional elements. Unlike the UBX proteins expressed in E. coli, UBX isoforms expressed in D. melanogaster cells are phosphorylated on serine and threonine residues, located primarily within a 53 aa region near the middle of the N-constant region, to form at least five phosphorylated states per isoform. Similar, if not identical states can be generated in vitro from purified E. coli UBX protein by a kinase activity in nuclear extracts from D. melanogaster cells. Temporal developmental profiles of UBX isoforms parallel those for the respective mRNAs, and all isoforms are similarly phosphorylated throughout embryogenesis. Analysis by cotransfection assays of the promoter activation and repression functions of mutant UBX proteins with various deletions in the N-constant region shows that repression is generally insensitive to deletion and, hence, presumably to phosphorylation. By contrast, the activation function is differentially sensitive to the different deletions in a manner indicating the absence of a discrete activating domain and instead, the presence of multiple activating sequences spread throughout the region.


Scientific and technical factors affecting the setting of Salmonella criteria for raw poultry: a global perspective.
Concerns about foodborne salmonellosis have led many countries to introduce microbiological criteria for certain food products. If such criteria are not well-grounded in science, they could be an unjustified obstacle to trade. Raw poultry products are an important part of the global food market. Import and export ambiguities and regulatory confusion resulting from different Salmonella requirements were the impetus for convening an international group of scientific experts from 16 countries to discuss the scientific and technical issues that affect the setting of a microbiological criterion for Salmonella contamination of raw chicken. A particular concern for the group was the use of criteria implying a zero tolerance for Salmonella and suggesting complete absence of the pathogen. The notion can be interpreted differently by various stakeholders and was considered inappropriate because there is neither an effective means of eliminating Salmonella from raw poultry nor any practical method for verifying its absence. Therefore, it may be more useful at present to set food safety metrics that involve reductions in hazard levels. Such terms as "zero tolerance" or "absence of a microbe" in relation to raw poultry should be avoided unless defined and explained by international agreement. Risk assessment provides a more meaningful approach than a zero tolerance philosophy, and new metrics, such as performance objectives that are linked to human health outcomes, should be utilized throughout the food chain to help define risk and identify ways to reduce adverse effects on public health.


The Influence of Clinical and Biological Factors on Transfusion-Associated Non-ABO Antigen Alloimmunization: Responders, Hyper-Responders, and Non-Responders.
In the context of transfusion medicine, alloimmunization most often refers to the development of antibodies to non-ABO red blood cell (RBC) antigens following pregnancy, transfusion, or transplantation. The development of RBC alloantibodies can have important clinical consequences, particularly in patients who require chronic transfusions. It has been suggested that alloimmunization is more common in some clinical circumstances and patient populations than in others. As such, individuals that develop alloantibodies are frequently referred to as 'responders' in the medical literature. In contrast, individuals that do not develop alloantibodies despite repeated exposures to non-self blood group antigens have been referred to as 'non-responders'. The purpose of this article is to review the phenomenon of RBC alloimmunization in the context of responders and non-responders to: i) establish a basic framework for alloimmunization as reported across several diverse patient populations; ii) more fully explore literature reports which support the concept of responders/non-responders regarding blood group antigen alloimmunization; iii) summarize the mechanisms that have been shown to predispose an individual to alloimmunization to determine how these factors may differentiate 'responders' from 'non-responders'; and iv) briefly discuss some practical approaches to prevent alloimmunization in patients who may be prone to alloantibody development.


Metal-free catalyst for the chemoselective methylation of amines using carbon dioxide as a carbon source.
N-methylation of amines is an important step in the synthesis of many pharmaceuticals and has been widely applied in the preparation of other key intermediates and chemicals. Therefore, the development of efficient methylation methods has attracted considerable attention. In this respect, carbon dioxide is an attractive C1 building block because it is an abundant, renewable, and nontoxic carbon source. Consequently, we developed a highly chemoselective, metal-free catalytic system that operates under ambient conditions for the N-methylation of amines.


Comparison of native and non-native phone imitation by English and Spanish speakers.
Experiments investigating phonetic convergence in conversation often focus on interlocutors with similar phonetic inventories. Extending these experiments to those with dissimilar inventories requires understanding the capacity of speakers to imitate native and non-native phones. In the present study, we tested native Spanish and native English speakers to determine whether imitation of non-native tokens differs qualitatively from imitation of native tokens. Participants imitated a [ba]-[pa] continuum that varied in VOT from -60 ms (prevoiced, Spanish [b]) to +60 ms (long lag, English [p]) such that the continuum consisted of some tokens that were native to Spanish speakers and some that were native to English speakers. Analysis of the imitations showed two critical results. First, both groups of speakers demonstrated sensitivity to VOT differences in tokens that fell within their native regions of the VOT continuum (prevoiced region for Spanish and long lag region for English). Secondly, neither group of speakers demonstrated such sensitivity to VOT differences among tokens that fell in their non-native regions of the continuum. These results show that, even in an intentional imitation task, speakers cannot accurately imitate non-native tokens, but are clearly flexible in producing native tokens. Implications of these findings are discussed with reference to the constraints on convergence in interlocutors from different linguistic backgrounds.


Mouse models in the era of large human tumour sequencing studies.
Cancer is a complex disease in which cells progressively accumulate mutations disrupting their cellular processes. A fraction of these mutations drive tumourigenesis by affecting oncogenes or tumour suppressor genes, but many mutations are passengers with no clear contribution to tumour development. The advancement of DNA and RNA sequencing technologies has enabled in-depth analysis of thousands of human tumours from various tissues to perform systematic characterization of their (epi)genomes and transcriptomes in order to identify (epi)genetic changes associated with cancer. Combined with considerable progress in algorithmic development, this expansion in scale has resulted in the identification of many cancer-associated mutations, genes and pathways that are considered to be potential drivers of tumour development. However, it remains challenging to systematically identify drivers affected by complex genomic rearrangements and drivers residing in non-coding regions of the genome or in complex amplicons or deletions of copy-number driven tumours. Furthermore, functional characterization is challenging in the human context due to the lack of genetically tractable experimental model systems in which the effects of mutations can be studied in the context of their tumour microenvironment. In this respect, mouse models of human cancer provide unique opportunities for pinpointing novel driver genes and their detailed characterization. In this review, we provide an overview of approaches for complementing human studies with data from mouse models. We also discuss state-of-the-art technological developments for cancer gene discovery and validation in mice.


Total and kallikrein arginine esterase activities in the urine of salt-hypertensive susceptible and resistant rats.
Urinary enzymes that hydrolyze the artificial substrate alpha-N-p-tosyl-L-arginine methyl ester (TAME) were studied in Dahl salt-sensitive (S) and salt-resistant (R) rats. Total urinary TAME esterase activity (kallikrein and non-kallikrein) showed a marked increase with dialysis against water, but only in hypertensive S rats with proteinuria. This phenomenon suggests the presence of dialyzable TAME esterase inhibitor(s) in urine following renal damage, but these data do not define what urinary esterases might be affected. Partially purified urinary kallikrein exhibited a ratio of kininogenase to esterase activity which was equal for S and R rats. Thus, the marked discrepancy between kininogenase and esterase activities reported by Carretero et al. with S and R whole urine is not a function of the S and R kallikrein molecules but is probably related to interfering substances in the whole urine. Urinary kallikrein excretion was measured on individual rat samples by TAME esterase activity following dialysis and separation from non-kallikrein TAME esterase(s) using DEAE-Sephadex minicolumns. S rats had lower urinary kallikrein excretion that R when the S rats were hypertensive and showed marked proteinuria. Young S and R rats raised on low salt showed similar blood pressures and similar kallikrein excretion. High salt (8% NaCl) diet decreased kallikrein excretion in both S and R, but the decrease was greater in the S rats which became hypertensive and had increased urine protein excretion. These data suggest that the lower urinary kallikrein excretion in S rats relative to R rats is probably a consequence of hypertension and renal damage rather than a primary cause of hypertension.


Intestinal organoid-based 2D monolayers mimic physiological and pathophysiological properties of the pig intestine.
Gastrointestinal infectious diseases remain an important issue for human and animal health. Investigations on gastrointestinal infectious diseases are classically performed in laboratory animals leading to the problem that species-specific models are scarcely available, especially when it comes to farm animals. The 3R principles of Russel and Burch were achieved using intestinal organoids of porcine jejunum. These organoids seem to be a promising tool to generate species-specific in vitro models of intestinal epithelium. 3D Organoids were grown in an extracellular matrix and characterized by qPCR. Organoids were also seeded on permeable filter supports in order to generate 2D epithelial monolayers. The organoid-based 2D monolayers were characterized morphologically and were investigated regarding their potential to study physiological transport properties and pathophysiological processes. They showed a monolayer structure containing different cell types. Moreover, their functional activity was demonstrated by their increasing transepithelial electrical resistance over 18 days and by an active glucose transport and chloride secretion. Furthermore, the organoid-based 2D monolayers were also confronted with cholera toxin derived from Vibrio cholerae as a proof of concept. Incubation with cholera toxin led to an increase of short-circuit current indicating an enhanced epithelial chloride secretion, which is a typical characteristic of cholera infections. Taken this together, our model allows the investigation of physiological and pathophysiological mechanisms focusing on the small intestine of pigs. This is in line with the 3R principle and allows the reduction of classical animal experiments.


Laparoendoscopic single-site nephrectomy using standard laparoscopic instruments: our initial experience.
To report our initial experience with laparoendoscopic single-site (LESS) nephrectomy using a GelPOINT single port and standard laparoscopic instruments. Laparoendoscopic single-site transperitoneal nephrectomy was done for 6 adult patients with a poorly functioning small or hydronephrotic kidney. The procedure was done with the GelPOINT system, which consisted of a wound retractor (Alexis) and GelSeal cap. Standard laparoscopic instruments were used and the renal pedicle was controlled with 10-mm Hem-o-Lok clips. The participants were 3 men and 3 women with the median age of 29.5 years. Laparoendoscopic single-site nephrectomy was successfully done in all the patients without any major complications. Median operation time was 110 minutes (range, 90 to 130 minutes). There was no need for blood transfusion in any patient. The recovery phase was uneventful and all the patients were discharged after a median hospital stay of 2.5 days (range, 2 to 3 days). Renal function remained stable in all the patients after the operation. The incision site healed well on postoperative follow-up. Our initial experience with LESS nephrectomy was successful with the use of a GelPOINT single port and standard laparoscopic instruments. This report may remove barriers to further work with the LESS technique and may offer a strategy to help surgeons gain experience with this novel technology.


Production of physician services under fee-for-service and blended fee-for-service: Evidence from Ontario, Canada.
We examine family physicians' responses to financial incentives for medical services in Ontario, Canada. We use administrative data covering 2003-2008, a period during which family physicians could choose between the traditional fee for service (FFS) and blended FFS known as the Family Health Group (FHG) model. Under FHG, FFS physicians are incentivized to provide comprehensive care and after-hours services. A two-stage estimation strategy teases out the impact of switching from FFS to FHG on service production. We account for the selection into FHG using a propensity score matching model, and then we use panel-data regression models to account for observed and unobserved heterogeneity. Our results reveal that switching from FFS to FHG increases comprehensive care, after-hours, and nonincentivized services by 3%, 15%, and 4% per annum. We also find that blended FFS physicians provide more services by working additional total days as well as the number of days during holidays and weekends. Our results are robust to a variety of specifications and alternative matching methods. We conclude that switching from FFS to blended FFS improves patients' access to after-hours care, but the incentive to nudge service production at the intensive margin is somewhat limited.


Assessment of antioxidant and antiproliferative properties of spinach plants grown under low oxygen availability.
In the human diet, the consumption of fresh fruits and vegetables is important in maintaining good health and in preventing chronic diseases. It is known that plant-derived food is a powerful source of chemopreventive molecules, i.e. antioxidants, and spinach (Spinacia oleracea L., Chenopodiaceae) possesses a wide range of metabolites with such biological activity. Plant stress response could lead to the production of metabolites with high value for human health and this could be a tool to enhance the production of molecules with antioxidant activity in plants. Data reported in this paper confirm the antioxidant properties of spinach plants, and show a strong antiproliferative activity of leaf extract on HT-29 human cell line. Besides, the hypoxic stress seems to affect the pool of antioxidant molecules present in spinach leaves, as verified by means of HPLC-MS/MS analysis and the aluminium chloride and ABTS assays. Our findings represent a basis for improving the biological and pharmacological properties of spinach plants, including the use of different growth conditions to modulate the phytocomplex profile of spinach.


94,000- and 100,000-molecular-weight simian virus 40 T-antigens are associated with the nuclear matrix in transformed and revertant mouse cells.
A small fraction of the 94,000-molecular-weight multifunctional large T-antigen of simian virus 40 was associated with the nuclear protein matrix derived from simian virus 40-transformed mouse cells. The interaction between this fraction of T-antigen and the matrix was largely or entirely independent of nuclear DNA. Similar amounts of T-antigen were retained by the nuclei of transformed and revertant cell lines. A 100,000-molecular-weight variant of T-antigen, which has been found to correlate specifically with anchorage-independent growth, was present in the nuclear protein matrix of a transformed cell line. A T-antigen-containing revertant selected for the reacquisition of a high serum requirement and an anchorage requirement for growth retained T-antigen in association with its matrix.


Oleander toxicosis in equids: 30 cases (1995-2010).
To determine clinical, laboratory analysis, and necropsy findings for equids with oleander toxicosis and to identify factors associated with outcome. Retrospective case series. 30 equids. Medical records of equids with detectable concentrations of oleandrin in serum, plasma, urine, or gastrointestinal fluid samples and equids that had not received cardiac glycoside drugs but had detectable concentrations of digoxin in serum were identified via a medical records database search. Descriptive statistics were calculated for medical history, physical examination, laboratory analysis, and necropsy variables. Logistic regression analysis was used to identify physical examination and laboratory analysis factors significantly associated with outcome. 3 of 30 (10.0%) equids died before or immediately after arrival at the hospital. Of the other 27 equids, 23 (85.2%) had gastrointestinal tract abnormalities, azotemia was detected for 19 (70.4%), and a cardiac arrhythmia was ausculted for 18 (66.7%). Mortality rate for all equids was 50.0%; mortality rate for hospitalized equids was 44.4%. The most common cause of death was cardiac dysfunction. Odds of survival to discharge from the hospital were lower for equids with cardiac arrhythmias versus those without arrhythmias and decreased with increasing Hct and serum glucose concentrations. Odds of survival increased with increasing serum chloride concentration and duration of hospitalization. Equids with oleander toxicosis frequently had simultaneous gastrointestinal tract, cardiac, and renal problems. Oleander intoxication should be a differential diagnosis for equids with colic in geographic areas where oleander is found, especially when azotemia or cardiac arrhythmias are detected concurrently.


Concise synthesis of complicated polypropionates through one-pot dissymmetrical two-directional chain elongation.
Herein, a major breakthrough in a sulfur dioxide-mediated oxyallylation cascade reaction is reported that allows the preparation of complex long-chain polyketide fragments with more than ten stereogenic centers through a carefully designed desymmetrization process. An allylbissilane is combined, under the appropriate reaction conditions, with two different 1,3-dioxy-1,3-dienes permitting the construction of a 13-membered polypropionate precursor in one pot. Four stereocenters are selectively created during this process. The so-obtained pseudo-C(2)- or -C(S)-symmetric products are desymmetrized through selective deprotection and can be selectively elongated in both directions using aldol chemistry.


SLAMF3 promotes Th17 differentiation and is reversed by iguratimod through JAK1/STAT3 pathway in primary Sjögren's syndrome.
The signaling lymphocytic activation molecule family of receptors (SLAMF) is involved in the activation of T cells and plays important roles in the pathogenesis of autoimmune diseases. The purpose of this study is to observe the expression of SLAMF3 on CD4 + T cells and its effect on the differentiation of T helper 17 (Th17) in primary Sjögren's syndrome (pSS). Furthermore, we found iguratimod (IGU) could effectively reverse the aberrant Th17 differentiation through JAK1/STAT3 signaling. Peripheral blood mononuclear cells from 40 pSS and 40 healthy control subjects were enrolled for analysis of expression of SLAMF3 on CD4 + T and Th17 cells by flow cytometry. Serum IL-17 and SLAMF3 were detected by ELISA assay. Labial biopsies from 20 pSS patients and 20 non-pSS controls were performed immunohistochemical for staining expression of CD4, IL-17, and SLAMF3. Under the priming conditions with anti-CD3/CD28 or CD3/SLAMF3 antibodies on CD4 + T cells extracted from pSS and controls, the proportion of Th17 cells in CD4 + T cells and the amount of soluble IL-17A were assessed by flow cytometry and ELISA. Furthermore, RNA sequencing was performed for the transcriptomics study. Additionally, RNA level of RORγt and IL-17A and the protein level of RORγt, p-JAK1 and p-STAT3, were detected by real-time PCR and western blot. The expression levels of SLAMF3 on CD4 + T and Th17 cells in the peripheral blood and salivary glands in pSS patients were significantly elevated than that in control groups. The serum IL-17A and SLAMF3 in pSS patients were much higher compared with the control group. Although co-stimulation of CD3/SLAMF3 could promote CD4 + T cells differentiate into Th17 cells both in pSS and controls, the CD4 + T cells from pSS have a more sensitive response in Th17 differentiation with the SLAMF3 stimulation. Transcriptomics results showed the CD3/SLAMF3 stimulation caused the activation of Th17 signaling and JAK1/STAT3 pathway. Quantitative PCR and western blotting confirmed the IGU (iguratimod), which is a safe clinical drug in treatment of autoimmune diseases, effectively reversed the increased Th17 proportion, the expression levels of RORγt, pJAK1, and pSTAT3 caused by CD3/SLAMF3 stimulation. SLAMF3 upregulates Th17 cell differentiation of CD4 + T cells and IL-17A secretion through enriching RORγt and activating the transcriptomics participating in the pathogenesis of primary Sjögren's syndrome. IGU could inhibit the process through this therapeutic target in pSS.


Characterization of a proteinase inhibitor from Brachypodium distachyon suggests the conservation of defence signalling pathways between dicotyledonous plants and grasses.
SUMMARY Proteinase inhibitors (PIs) are established markers for wound- and especially jasmonate-mediated signalling in dicot species such as tomato and potato. Differential screening of a cDNA library constructed from RNA isolated from wounded leaves of the grass Brachypodium distachyon led to the identification of a proteinase inhibitor gene (Bdpin1). Bdpin1 exhibited the highest homology to the subtilisin/chymotrypsin-inhibiting subgroup of the pin1 class of plant PIs. Northern analyses indicated that Bdpin1 was induced within 6 h at the site of wounding and systemically, by 24 h, thereby providing evidence for long-distance signalling in grasses. Bdpin1 also proved to be more rapidly induced in susceptible than in resistant ecotypes of B. distachyon following challenge with the Rice blast pathogen, Magnaporthe grisea. Screening with chemical signals indicated that Bdpin1 could be induced with MeJA but not with the putative mimic of salicylic acid, benzothiadiazole. Genomic Southern hybridization was consistent with Bdpin1 existing at a single locus, which was isolated following screening of a genomic cosmid library. DNA upstream of the Bdpin1 coding sequence was characterized via fusion to a GUS reporter and was found to confer wound-responsive transcription in B. distachyon and other cereals following biolistic bombardment. Both wound- and TMV-activated Bdpin1-GUS activity was detected in transgenic tobacco. Given that B. distachyon represents an ancestral grass species, our data suggest that there is considerable conservation in defence-associated signalling between dicots and grasses.


Removal of cactus spines from the skin. A comparative evaluation of several methods.
The removal of very fine cactus spines from the skin is particularly distressing for the pediatric patient. We describe two typical patients and a study in experimental animals comparing the effectiveness of several previously described methods for removal. The most effective method involved using tweezers to remove clumps of spines followed by a thin layer of glue covered with gauze, which was allowed to dry and then peeled off to remove individual spines. Attempts to use adhesive tape or a thin layer of a commercial facial mask to aid in removal of the spines produced more retention and inflammation three days after removal than no treatment.


Genistein inhibits proliferation and induces senescence in neonatal mouse pituitary gland explant cultures.
Genistein is an isoflavone abundant in soybean and infants are exposed to high levels of genistein in soy-based formula. It is known that genistein mediates estrogen receptor (ER) signaling, and exposure during neonatal development could cause acute and long term endocrine effects. We assayed genistein's impact on the neonatal mouse pituitary gland because it is an endocrine signaling hub and is sensitive to endocrine disruption during critical periods. Pituitary explant cultures, which actively proliferate and differentiate, were exposed to 0.06 μM-36 μM genistein and assayed for mRNA and protein changes. Genistein induced mRNA expression of the ERα regulated gene, Cckar, to the same magnitude as estradiol (E2) but with less potency. Interestingly, 36 μM genistein strongly inhibited pituitary proliferation, measured by a reduction in mKi67 mRNA and phospho-Histone H3 immunostaining. Examining cell cycle dynamics, we found that 36 μM genistein decreased Ccnb1 (Cyclin B1) mRNA; while mRNA for the cyclin dependent kinase inhibitor Cdkn1a (p21) was upregulated, correlated with an apparent increase in p21 immunostained cells. Strikingly, we observed a robust onset of cellular senescence, permanent cell cycle exit, in 36 μM genistein treated pituitaries by increased senescence activated β-galactosidase staining. We also found that 36 μM genistein decreased Bcl2 mRNA levels, a gene protective against apoptosis. Taken together these data suggest that genistein exposure during the neonatal period could initiate senescence and halt proliferation during a time when the proper numbers of endocrine cells are being established for mature gland function.


Cell shape impacts on the positioning of the mitotic spindle with respect to the substratum.
All known mechanisms of mitotic spindle orientation rely on astral microtubules. We report that even in the absence of astral microtubules, metaphase spindles in MDCK and HeLa cells are not randomly positioned along their x-z dimension, but preferentially adopt shallow β angles between spindle pole axis and substratum. The nonrandom spindle positioning is due to constraints imposed by the cell cortex in flat cells that drive spindles that are longer and/or wider than the cell's height into a tilted, quasidiagonal x-z position. In rounder cells, which are taller, fewer cortical constraints make the x-z spindle position more random. Reestablishment of astral microtubule-mediated forces align the spindle poles with cortical cues parallel to the substratum in all cells. However, in flat cells, they frequently cause spindle deformations. Similar deformations are apparent when confined spindles rotate from tilted to parallel positions while MDCK cells progress from prometaphase to metaphase. The spindle disruptions cause the engagement of the spindle assembly checkpoint. We propose that cell rounding serves to maintain spindle integrity during its positioning.


Prefrontal Asymmetry BCI Neurofeedback Datasets.
Prefrontal cortex (PFC) asymmetry is an important marker in affective neuroscience and has attracted significant interest, having been associated with studies of motivation, eating behavior, empathy, risk propensity, and clinical depression. The data presented in this paper are the result of three different experiments using PFC asymmetry neurofeedback (NF) as a Brain-Computer Interface (BCI) paradigm, rather than a therapeutic mechanism aiming at long-term effects, using functional near-infrared spectroscopy (fNIRS) which is known to be particularly well-suited to the study of PFC asymmetry and is less sensitive to artifacts. From an experimental perspective the BCI context brings more emphasis on individual subjects' baselines, successful and sustained activation during epochs, and minimal training. The subject pool is also drawn from the general population, with less bias toward specific behavioral patterns, and no inclusion of any patient data. We accompany our datasets with a detailed description of data formats, experiment and protocol designs, as well as analysis of the individualized metrics for definitions of success scores based on baseline thresholds as well as reference tasks. The work presented in this paper is the result of several experiments in the domain of BCI where participants are interacting with continuous visual feedback following a real-time NF paradigm, arising from our long-standing research in the field of affective computing. We offer the community access to our fNIRS datasets from these experiments. We specifically provide data drawn from our empirical studies in the field of affective interactions with computer-generated narratives as well as interfacing with algorithms, such as heuristic search, which all provide a mechanism to improve the ability of the participants to engage in active BCI due to their realistic visual feedback. Beyond providing details of the methodologies used where participants received real-time NF of left-asymmetric increase in activation in their dorsolateral prefrontal cortex (DLPFC), we re-establish the need for carefully designing protocols to ensure the benefits of NF paradigm in BCI are enhanced by the ability of the real-time visual feedback to adapt to the individual responses of the participants. Individualized feedback is paramount to the success of NF in BCIs.