Preoperative intravenous fluid therapy decreases postoperative nausea and pain in high risk patients.
The potential for preoperative IV rehydration to reduce postoperative nausea and vomiting (PONV) and pain in patients undergoing ambulatory surgery remains unclear, with conflicting results reported. We sought to determine whether preoperative IV rehydration with a balanced salt solution would decrease the incidence of PONV in patients at increased risk for these symptoms. Eighty ASA grade I-III patients presenting for gynecologic laparoscopy were randomized to receive large (2 mL/kg per hour fasting) or small (3 mL/kg) volume infusions of compound sodium lactate solution over 20 min preoperatively. A standardized balanced anesthetic was used. The incidence and severity of PONV and pain, and need for supplemental antiemetic and analgesic therapy, were assessed by a blinded investigator at 0.5, 1, and 4 h postoperatively, and on the first and third postoperative days. The incidence (control 87% versus large volume 59%) and severity of PONV were significantly reduced in the large volume infusion group at all time intervals. The large volume infusion group also had decreased postoperative pain scores and required less supplemental analgesia. Preoperative correction of intravascular volume deficits effectively reduces PONV and postoperative pain in high risk patients presenting for ambulatory surgery. We recommend the preoperative administration of 2 mL/kg of compound sodium lactate for every hour of fasting to patients with an increased PONV risk presenting for ambulatory surgery.


The relation between problem categorization and problem solving among experts and novices.
These investigations were conducted to examine the relationship between problem-solving ability and the criteria used to decide that two classical mechanics problems would be solved similarly. We began by comparing experts and novices on a similarity judgment task and found that the experts predominantly relied on the problems' deep structures in deciding on similarity of solution, although the presence of surface-feature similarity had a clear adverse effect on performance. The novices relied predominantly on surface features, but were capable of using the problems' deep structures under certain conditions. In a second experiment, we compared groups of novices, at the same level of experience, who tended to employ different types of reasoning in making similarity judgments. Compared to novices who relied predominantly on surface features, novices who made greater use of principles tended to categorize problems similarly to how experts categorized them, as well as score higher in problem solving. These results suggest that principles play a fundamental role in the organization of conceptual and procedural knowledge for good problem solvers at all levels.


Unplanned complex suicide by self-strangulation associated with multiple sharp force injuries: a case report.
In cases of ligature strangulation, the importance of distinguishing self-inflicted death from homicide is crucial. This entails objective scene investigation, autopsy and anamnesis in order to elucidate the manner of death correctly. The authors report a case of unplanned complex suicide by means of self-strangulation and multiple sharp force injury. The use of more than one suicide method, consecutively--termed unplanned complex suicide--gives this case particular significance. A brief discussion on this uncommon method of suicide is presented, particularly relevant to the attending forensic physician. In addition, a short overview of the entity of complex suicide is given.


What traits are carried on mobile genetic elements, and why?
Although similar to any other organism, prokaryotes can transfer genes vertically from mother cell to daughter cell, they can also exchange certain genes horizontally. Genes can move within and between genomes at fast rates because of mobile genetic elements (MGEs). Although mobile elements are fundamentally self-interested entities, and thus replicate for their own gain, they frequently carry genes beneficial for their hosts and/or the neighbours of their hosts. Many genes that are carried by mobile elements code for traits that are expressed outside of the cell. Such traits are involved in bacterial sociality, such as the production of public goods, which benefit a cell's neighbours, or the production of bacteriocins, which harm a cell's neighbours. In this study we review the patterns that are emerging in the types of genes carried by mobile elements, and discuss the evolutionary and ecological conditions under which mobile elements evolve to carry their peculiar mix of parasitic, beneficial and cooperative genes.


Activation of preformed EGF receptor dimers by ligand-induced rotation of the transmembrane domain.
The epidermal growth factor receptor plays crucial roles throughout the development of multicellular organisms, and inappropriate activation of the receptor is associated with neoplastic transformation of many cell types. The receptor is thought to be activated by ligand-induced homodimerisation. Here, however, we show by chemical cross-linking and sucrose density-gradient centrifugation that in the absence of bound ligand the receptor has an ability to form a dimer and exists as a preformed dimer on the cell surface. We also analysed the receptor dimerisation by inserting cysteine residues at strategic positions about the putative alpha-helix axis of the extracellular juxtamembrane region. The mutant receptors spontaneously formed disulphide bridges and transformed NIH3T3 cells in the absence of ligand, depending upon the positions of the cysteine residue inserted. Kinetic analyses of the disulphide bonding indicate that EGF binding induces flexible rotation or twist of the juxtamembrane region of the receptor in the plane parallel with the lipid bilayer. The binding of an ATP competitor to the intracellular domain also induced similar flexible rotation of the juxtamembrane region. All the disulphide-bonded dimers had flexible ligand-binding domains with the same biphasic affinities for EGF as the wild-type. These results demonstrate that ligand binding to the flexible extracellular domains of the receptor dimer induce rotation or twist of the juxtamembrane regions, hence the transmembrane domains, and dissociate the dimeric, inactive form of the intracellular domains. The flexible rotation of the intracellular domains may be necessary for the intrinsic catalytic kinase to become accessible to the multiple tyrosine residues present in the regulatory domain and various substrates, and may be a common property of many cell-surface receptors, such as the insulin receptor.


Bipolar risk and mental imagery susceptibility in a representative sample of Chinese adults residing in the community.
We need to better understand the cognitive factors associated with risk for bipolar disorders. Recent research suggests that increased susceptibility to mental imagery may be one such factor. However, since this research was primarily conducted with Western students and at a single time-point, it is not known whether the relationship between imagery susceptibility and bipolar symptoms exists across cultures or within the general community, or whether this relationship remains stable over time. This study evaluated whether Chinese adults identified as being at high (HR) versus low (LR) risk of developing bipolar disorders showed greater mental imagery susceptibility. We aimed to test whether such a relationship was stable over time by measuring imagery characteristics at baseline and at the 7-week follow-up. This prospective study recruited a community sample of N = 80 Chinese adults screened for the absence of neurotic and psychotic disorders. The sample was split into HR (n = 18) and LR (n = 62) groups at baseline based on a criterion cut-off score on a measure of hypomania, the Mood Disorder Questionnaire (MDQ). Participants completed measures of imagery susceptibility and its impact: the Spontaneous Use of Imagery Scale (SUIS) and the Impact of Future Events Scale (IFES), at baseline and 7 weeks later. HR group reported greater tendency to use imagery in daily life (SUIS) and greater emotional impact of prospective imagery (IFES) than LR group at baseline. These results remained stable at follow-up. This study provides preliminary evidence for increased susceptibility to mental imagery in individuals at high risk of bipolar disorders recruited from a community sample of Chinese adults. This extends previous research in Western student samples suggesting that imagery (both levels of use and its emotional impact) may be a cognitive factor with cross-cultural relevance that is stable over time.


Inducible nitric oxide synthase (iNOS) in sinonasal polyp pathogenesis.
We investigated the role of inducible nitric oxide synthase (iNOS) in the pathogenesis of sinonasal polyps. Adult patients (21 men, 3 women) with nasal polyposis underwent functional endoscopic sinus surgery. Nine adults without polyps (6 men) who underwent septoplasty and/or rhinoplasty served as controls. Polyp specimens came from three regions: the maxillary sinus (10), ethmoid sinus (14), and nasal cavity (10). Control group samples (9) came from the inferior turbinate. Specimens were evaluated in eight mucosal layers for count and distribution of inflammatory cells and iNOS expression. An iNOS positivity index (PI) was determined for the epithelium (E), subepithelial layer of the lamina propria (SE), and deep paraglandular layer of the mucosa (D). Polymorphonuclear cell (PMNC) % values of the ethmoid and maxillary sinus and overall ethmoid sinus PI were significantly higher in the polyp group. Patients with longer polyp duration, D-perivascular (D-pv), and a higher Brinkmann index had decreased ethmoid sinus D PIs. However, in older patients and patients with longer polyp duration, perivascular PIs increased in maxillary sinus SE and D, respectively. Furthermore, as PMNC % and iNOS-PMNC PI increased, SE_glandular and epithelial_apical iNOS values decreased. In the ethmoid and maxillary sinuses, iNOS_D_. endothelial values increased but decreased in the nasal cavity. iNOS may play a role in sinonasal polyp pathogenesis, especially in mucosal SE and D layers. Increased vascular permeability, stromal edema, inflammatory cell migration into the stroma of the mucosa, and increased mucosal gland secretion may result in polyp formation.


Maternal ethanol consumption alters the epigenotype and the phenotype of offspring in a mouse model.
Recent studies have shown that exposure to some nutritional supplements and chemicals in utero can affect the epigenome of the developing mouse embryo, resulting in adult disease. Our hypothesis is that epigenetics is also involved in the gestational programming of adult phenotype by alcohol. We have developed a model of gestational ethanol exposure in the mouse based on maternal ad libitum ingestion of 10% (v/v) ethanol between gestational days 0.5-8.5 and observed changes in the expression of an epigenetically-sensitive allele, Agouti viable yellow (A(vy)), in the offspring. We found that exposure to ethanol increases the probability of transcriptional silencing at this locus, resulting in more mice with an agouti-colored coat. As expected, transcriptional silencing correlated with hypermethylation at A(vy). This demonstrates, for the first time, that ethanol can affect adult phenotype by altering the epigenotype of the early embryo. Interestingly, we also detected postnatal growth restriction and craniofacial dysmorphology reminiscent of fetal alcohol syndrome, in congenic a/a siblings of the A(vy) mice. These findings suggest that moderate ethanol exposure in utero is capable of inducing changes in the expression of genes other than A(vy), a conclusion supported by our genome-wide analysis of gene expression in these mice. In addition, offspring of female mice given free access to 10% (v/v) ethanol for four days per week for ten weeks prior to conception also showed increased transcriptional silencing of the A(vy) allele. Our work raises the possibility of a role for epigenetics in the etiology of fetal alcohol spectrum disorders, and it provides a mouse model that will be a useful resource in the continued efforts to understand the consequences of gestational alcohol exposure at the molecular level.


Occupational social class, risk factors and cardiovascular disease incidence in men and women: a prospective study in the European Prospective Investigation of Cancer and Nutrition in Norfolk (EPIC-Norfolk) cohort.
To investigate the association between occupational social class and cardiovascular disease (CVD) incidence, and the extent to which classical and lifestyle risk factors explain such relationships, and if any differences persist after 65 years of age. Prospective population study of 22,478 men and women aged 39-79 years living in the general community in Norfolk, United Kingdom, recruited using general practice age-sex registers in 1993-1997 and followed up for total mortality to 2006. In both men and women an inverse relationship was observed between social class and CVD incidence, with a relative risk of social class V compared to I of 1.90 in men (95% CI 1.47 to 2.47, P < 0.001) and 1.90 in women (95% CI 1.45 to 2.49, P < 0.001). Adjusting for classical and lifestyle risk factors (age, smoking, BMI, systolic blood pressure, total blood cholesterol, history of diabetes, physical activity, weekly alcohol intake and plasma vitamin C levels) had little effect in men; the relative risk of social class V compared to I of 1.70 (95% CI 1.31 to 2.22, P < 0.001), while there was some attenuation seen in women, relative risk of social class V compared to I of 1.56 (95% CI 1.18 to 2.05, P = 0.011). The association persisted in men and women aged > or =65 years. Some but not all of the socioeconomic differential in CVD incidence can be explained by potentially modifiable classical and lifestyle risk factors. Low social class remains a risk factor for CVD after age 65 years. Further understanding of the mechanisms underlying the association is needed if we are to reduce inequalities in health.


Activation of protein kinase C-ζ in pancreatic β-cells in vivo improves glucose tolerance and induces β-cell expansion via mTOR activation.
PKC-ζ activation is a key signaling event for growth factor-induced β-cell replication in vitro. However, the effect of direct PKC-ζ activation in the β-cell in vivo is unknown. In this study, we examined the effects of PKC-ζ activation in β-cell expansion and function in vivo in mice and the mechanisms associated with these effects. We characterized glucose homeostasis and β-cell phenotype of transgenic (TG) mice with constitutive activation of PKC-ζ in the β-cell. We also analyzed the expression and regulation of signaling pathways, G1/S cell cycle molecules, and β-cell functional markers in TG and wild-type mouse islets. TG mice displayed increased plasma insulin, improved glucose tolerance, and enhanced insulin secretion with concomitant upregulation of islet insulin and glucokinase expression. In addition, TG mice displayed increased β-cell proliferation, size, and mass compared with wild-type littermates. The increase in β-cell proliferation was associated with upregulation of cyclins D1, D2, D3, and A and downregulation of p21. Phosphorylation of D-cyclins, known to initiate their rapid degradation, was reduced in TG mouse islets. Phosphorylation/inactivation of GSK-3β and phosphorylation/activation of mTOR, critical regulators of D-cyclin expression and β-cell proliferation, were enhanced in TG mouse islets, without changes in Akt phosphorylation status. Rapamycin treatment in vivo eliminated the increases in β-cell proliferation, size, and mass; the upregulation of cyclins Ds and A in TG mice; and the improvement in glucose tolerance-identifying mTOR as a novel downstream mediator of PKC-ζ-induced β-cell replication and expansion in vivo. CONCLUSIONS PKC:-ζ, through mTOR activation, modifies the expression pattern of β-cell cycle molecules leading to increased β-cell replication and mass with a concomitant enhancement in β-cell function. Approaches to enhance PKC-ζ activity may be of value as a therapeutic strategy for the treatment of diabetes.


Binocular depth reversals despite familiarity cues.
Stereoscopic depth can be reversed by interchanging the left- and right-eye views (pseudoscopy) when abstract stereograms are used, but not when stereograms contain natural objects or scenes. This resistance to reversal of depth has traditionally been attributed to familiarity with the shape of objects and the presence of monocular depth cues. However, when texture disparity is neutralized by making the texture perspective of surfaces identical for both eyes, even a highly familiar object, like a monocularly recognizable human face, appears as concave (nose pointing inwards) when viewed pseudoscopically.


A caged imidazopyrazinone for selective bioluminescence detection of labile extracellular copper(ii).
Copper is an essential redox-active metal that plays integral roles in biology ranging from enzymatic catalysis to mitochondrial respiration. However, if not adequately regulated, this redox activity has the potential to cause oxidative stress through the production of reactive oxygen species. Indeed, the dysregulation of copper has been associated with a variety of disease states including diabetes, neurodegenerative disorders, and multiple cancers. While increasing tools are being developed for illuminating labile intracellular copper pools and the trafficking pathways in which they are involved, significantly less attention has been given to the analogous extracellular labile pool. To address this gap, we have developed a bioluminescence-based imaging probe, picolinic ester caged-diphenylterazine (pic-DTZ) for monitoring labile, extracellular copper using a coelenterazine-like imidazopyrazinone and the genetically-engineered, marine-based luciferase, nanoluciferase. Unlike the more commonly-used firefly luciferase, nanoluciferase does not require ATP, allowing its application to the extracellular milieu. pic-DTZ demonstrates high metal and oxidation state selectivity for Cu(ii) in aqueous buffer as well as selectivity for labile pools over coordinatively inaccessible protein-bound Cu(ii). We demonstrate the potential of pic-DTZ as a diagnostic tool in human serum and plasma for copper-associated diseases. Additionally, we apply pic-DTZ to lend insight into the extracellular copper dynamic in anticancer treatments.


A four-subunit DNA polymerase ζ complex containing Pol δ accessory subunits is essential for PCNA-mediated mutagenesis.
DNA polymerase ζ (Pol ζ) plays a key role in DNA translesion synthesis (TLS) and mutagenesis in eukaryotes. Previously, a two-subunit Rev3-Rev7 complex had been identified as the minimal assembly required for catalytic activity in vitro. Herein, we show that Saccharomyces cerevisiae Pol ζ binds to the Pol31 and Pol32 subunits of Pol δ, forming a four-subunit Pol ζ(4) complex (Rev3-Rev7-Pol31-Pol32). A [4Fe-4S] cluster in Rev3 is essential for the formation of Pol ζ(4) and damage-induced mutagenesis. Pol32 is indispensible for complex formation, providing an explanation for the long-standing observation that pol32Δ strains are defective for mutagenesis. The Pol31 and Pol32 subunits are also required for proliferating cell nuclear antigen (PCNA)-dependent TLS by Pol ζ as Pol ζ(2) lacks functional interactions with PCNA. Mutation of the C-terminal PCNA-interaction motif in Pol32 attenuates PCNA-dependent TLS in vitro and mutagenesis in vivo. Furthermore, a mutant form of PCNA, encoded by the mutagenesis-defective pol30-113 mutant, fails to stimulate Pol ζ(4) activity, providing an explanation for the observed mutagenesis phenotype. A stable Pol ζ(4) complex can be identified in all phases of the cell cycle suggesting that this complex is not regulated at the level of protein interactions between Rev3-Rev7 and Pol31-Pol32.


Evaluation of User-Interface Alert Displays for Clinical Decision Support Systems for Sepsis.
Hospitals are increasingly turning to clinical decision support systems for sepsis, a life-threatening illness, to provide patient-specific assessments and recommendations to aid in evidence-based clinical decision-making. Lack of guidelines on how to present alerts has impeded optimization of alerts, specifically, effective ways to differentiate alerts while highlighting important pieces of information to create a universal standard for health care providers. To gain insight into clinical decision support systems-based alerts, specifically targeting nursing interventions for sepsis, with a focus on behaviors associated with and perceptions of alerts, as well as visual preferences. An interactive survey to display a novel user interface for clinical decision support systems for sepsis was developed and then administered to members of the nursing staff. A total of 43 nurses participated in 2 interactive survey sessions. Participants preferred alerts that were based on an established treatment protocol, were presented in a pop-up format, and addressed the patient's clinical condition rather than regulatory guidelines. The results can be used in future research to optimize electronic medical record alerting and clinical practice workflow to support the efficient, effective, and timely delivery of high-quality care to patients with sepsis. The research also may advance the knowledge base of what information health care providers want and need to improve the health and safety of their patients.


A systematic review and quality analysis of pediatric traumatic brain injury clinical practice guidelines.
Traumatic brain injuries (TBI) are a significant cause of mortality and morbidity for children globally. Adherence to evidence-based treatment guidelines have been shown to improve TBI outcomes. To inform the creation of a pediatric TBI management guideline for a low and middle income country context, we assessed the quality of available clinical practice guidelines (CPGs) for the acute management pediatric TBI. Articles were identified and retrieved from MEDLINE, EMBASE, Cochrane Library, LILACS, Africa-Wide Information and Global Index Medicus. These articles were screened by four reviewers independently. Based on the eligibility criteria, with the exception of literature reviews, opinion papers and editor's letters, articles published from 1995 to November 11, 2016 which covered clinical recommendations, clinical practice or treatment guidelines for the acute management of pediatric TBI (within 24 hours) were included for review. A reference and citation analysis was performed. Seven independent reviewers from low, middle and high income clinical settings with knowledge of pediatric TBI management appraised the guidelines using the AGREE II instrument. Scores for the CPGs were aggregated by domain and overall assessment was determined. We screened 2372 articles of which 17 were retained for data extraction and guideline appraisal. Except for one CPG from a middle income country, the majority (16/17) of the guidelines were developed in high income countries. Seven guidelines were developed specifically for the pediatric population, while the remaining CPGs addressed the acute management of TBI in both adult and pediatric populations. The New Zealand Guideline Group (NZGG, 2006) received the highest overall assessment score of 46/49 (93.88%) followed by the Scandinavian Neurotrauma Committee (SNC, 2016) with a score of 45/49 (91.84%) followed by the Scottish Intercollegiate Guideline Network (SIGN, 2009) and Brain Trauma Foundation (BTF 2012) both with scores of 44/49 (89.80%). CPGs from Cincinnati Children's Hospital (CCH 2006) and Sao Paulo Medical School Hospital/Brazilian Society of Neurosurgery (USP/BSN, 2001) received the lowest score of 27/49 (55.10%) subsequently followed by the Appropriateness Criteria (ACR, 2015) with 29/49 (59.18%). The domains for scope and purpose and clarity of presentation received the highest scores across the CPGs, while applicability and editorial independence domains had the lowest scores with a wider variability in score range for rigor of development and stakeholder involvement. To our knowledge, this is the first systematic review and guideline appraisal for pediatric CPGs concerning the acute management of TBI. Targeted guideline creation specific to the pediatric population has the potential to improve the quality of acute TBI CPGs. Furthermore, it is crucial to address the applicability of a guideline to translate the CPG from a published manuscript into clinically relevant local practice tools and for resource limited practice settings.


Reexamining Dis/Similarity-Based Tests for Rare-Variant Association with Case-Control Samples.
A properly designed distance-based measure can capture informative genetic differences among individuals with different phenotypes and can be used to detect variants responsible for the phenotypes. To detect associated variants, various tests have been designed to contrast genetic dissimilarity or similarity scores of certain subject groups in different ways, among which the most widely used strategy is to quantify the difference between the within-group genetic dissimilarity/similarity (i.e., case-case and control-control similarities) and the between-group dissimilarity/similarity (i.e., case-control similarities). While it has been noted that for common variants, the within-group and the between-group measures should all be included; in this work, we show that for rare variants, comparison based on the two within-group measures can more effectively quantify the genetic difference between cases and controls. The between-group measure tends to overlap with one of the two within-group measures for rare variants, although such overlap is not present for common variants. Consequently, a dissimilarity or similarity test that includes the between-group information tends to attenuate the association signals and leads to power loss. Based on these findings, we propose a dissimilarity test that compares the degree of SNP dissimilarity within cases to that within controls to better characterize the difference between two disease phenotypes. We provide the statistical properties, asymptotic distribution, and computation details for a small sample size of the proposed test. We use simulated and real sequence data to assess the performance of the proposed test, comparing it with other rare-variant methods including those similarity-based tests that use both within-group and between-group information. As similarity-based approaches serve as one of the dominating approaches in rare-variant analysis, our results provide some insight for the effective detection of rare variants.


Low-input Capture-C: A Chromosome Conformation Capture Assay to Analyze Chromatin Architecture in Small Numbers of Cells.
Chromosome conformation capture (3C) techniques are crucial to understanding tissue-specific regulation of gene expression, but current methods generally require large numbers of cells. This protocol describes two new low-input Capture-C approaches that can generate high-quality 3C interaction profiles from 10,000-20,000 cells, depending on the resolution used for analysis.


The Availability of Supervision in Routine Mental Health Care.
Clinical supervision is an embedded resource for practice quality in community mental health organizations. Supervision has been found to increase provider competence and decrease stress. In addition, supervision has been associated with service user outcomes including decreased depressive symptoms. However, little is known about the availability and nature of supervision in real world settings. The primary aims of this study were to identify available supervision and the extent to which contextual factors are related to that availability. The data source for this study was a multi-state and multi-site (N=14) NIMH-funded trial survey of providers (N=273). Supervision was measured by hours per week (quantity) and by utilization of best practice activities (content). Univariate, chi-square, independent samples t-tests, and ANOVA analyses were used to assess supervision content and quantity and to examine subgroup differences. Participants reported an average of 2.17 hours of supervision per week and 28.6% of participants endorsed best practice content. Supervision quantity varied significantly across sites (p<.05) and program type (p<.05) while content did not. Individual role within the organization had a significant relationship with reported supervision content (p<.001). In these settings, organizations are exercising discretion in how to utilize supervision within the available time. Supervision time also varied by program type, increasing with the intensity of services. Findings demonstrate that reports of availability vary according to position within the organization and the intensity of services within a given program type. Implications for workforce development, access to quality services, and implementation of evidence-based practices are discussed.


Psychometric properties of the Swedish version of the burn outcomes questionnaire for children aged 5 to 18 years.
Although pediatric burn injuries are common, there is a lack of burn-specific health outcome measurements for children. The American Burn Association and the Shriners Hospitals for Children have developed the Burn Outcomes Questionnaire (BOQ), which is a parent-report questionnaire measuring the functional outcome after burn in children aged 5 to 18 years. The aim of this study was to examine the psychometric properties of the Swedish version of the BOQ, assessing feasibility, reliability, and validity aspects. Participants were parents (n = 70) of children aged 5 to 18 years who were treated at the Uppsala or Linköping burn center between January 2000 and December 2008. For most subscales, feasibility was adequate and the internal consistency was good: Cronbach's α values were above 0.76 in all but 1 subscale, and mean interitem correlations ranged from 0.34 to 0.90. The test-retest reliability was significant in the majority of subscales. Evidence of validity was shown by associations among the BOQ subscales and between BOQ subscales and measures of burn severity, heat sensitivity, fear-avoidance beliefs, and parent reports of the child's psychological problems. In conclusion, with the exception of a few subscales, this study supports the continued evaluation of the Swedish version of BOQ as a tool to measure outcome after burn in children aged 5 to 18 years.


Clinical outcomes and predictors for ESRD and mortality in primary GN.
Relatively little is known about the long-term outcomes of different histologic types of primary glomerulonephritis in Asian populations. From 1993 to 2006, 987 patients undergoing renal biopsy were studied, and 580 patients (mean age=44.4 years, male=58.5%) with the four most common forms of glomerulonephritis (membranous nephropathy, focal and segmental glomerulosclerosis, IgA nephropathy, and minimal change disease) were selected for analysis. Median follow-up period was 5.9 (interquartile range=5.7) years. The focal and segmental glomerulosclerosis group displayed the highest incidence of ESRD (25.8%) and the fastest decline of estimated GFR (4.6 ml/min per 1.73 m(2) per year). The IgA nephropathy group also had a higher rate of ESRD than the membranous nephropathy patients (19.2% versus 4.3%, P<0.001). In contrast, the membranous nephropathy group exhibited an overall death rate similar to the focal and segmental glomerulosclerosis group (17.2% versus 14.4%) but higher than the IgA nephropathy and minimal change disease patients (4.6% and 3.7%, respectively, P<0.001). The most powerful predictor for ESRD was focal and segmental glomerulosclerosis, whereas the strongest predictor for all-cause mortality was membranous nephropathy with higher proteinuria. Protectors against ESRD included male sex and higher hemoglobin. Most predictors for ESRD and overall mortality found in this ethnic Chinese cohort were similar to other studies. However, some risk factors linked with distinct glomerular pathologies displayed differential clinical outcomes.


Metachronous Ovarian Metastases in a Patient with Primary Colorectal Cancer. A Case Report and Review of the Literature.
BACKGROUND Metachronous ovarian metastasis from primary colorectal cancer (CRC) is a rare condition that is diagnosed after the treatment of CRC. In most cases, ovarian metastases present without specific symptoms or signs and are usually diagnosed during follow-up imaging. A rare case is presented of metachronous ovarian metastasis from primary CRC, diagnosed on follow-up by computed tomography (CT) and magnetic resonance imaging (MRI), and includes a review of the literature. CASE REPORT A 66-year-old woman recently underwent a left hemicolectomy for a stage T3, N0, M0 primary adenocarcinoma of the sigmoid colon, which was completely excised. Three years later, follow-up CT and MRI imaging showed a right ovarian cyst. She underwent exploratory laparotomy and bilateral salpingo-oophorectomy, which identified tumor in the right ovary. Histopathology and immunohistochemistry confirmed metachronous ovarian metastasis from CRC. The patient was referred for further treatment. CONCLUSIONS Newly-diagnosed ovarian metastasis from primary colorectal cancer (CRC) is challenging to diagnose and manage, and may initially be incorrectly diagnosed as malignancy of primary ovarian origin. This case demonstrated that it is important to confirm the diagnosis with imaging, histology, and the appropriate use of tumor markers. Because ovarian metastases do not respond favorably to chemotherapy, the treatment of choice is surgery. However, for women who are treated for CRC, the use of prophylactic oophorectomy remains controversial.


Mitochondrial DNA sequence variation and phylogeographical structure of rock partridge (Alectoris graeca) populations.
The rock partridge (Alectoris graeca) is distributed in the Alps, Apennines and Balkans, in mountainous areas that were heavily affected by cyclic climate and landscape changes during the last Pleistocene glaciations. Some partridge populations have colonized and expanded their present ranges only after deglaciation and recent deforestation by humans. Other populations have been segregated in land-bridge islands isolated after the rise in level of the Mediterranean Sea. Consequently, partridges from different areas could be genetically differentiated. This study has analysed 436 nucleotides of the domain I of the mitochondrial DNA control-region in 70 rock partridges collected from 14 populations throughout their distribution. Phylogenetic analysis and analysis of molecular variance showed that the Sicilian population is very divergent from the continental populations. The Albanian and central Apennine samples are slightly divergent from the Alpine rock partridges, which cluster in two groups according to their east-west distributions. The mitochondrial DNA sequences suggest the existence of a phylogeographical structuring among rock partridge populations, resulting from genetic divergence in southern refugia and subsequent postglacial colonization of northern mountain areas. Sicilian partridges could have been isolated for approximately 500,000 years, whereas the Albanian-Apennine populations could have been in contact since the last glacial maximum (21,000 yr BP) through the north Adriatic land-bridge. The Alps could have been colonized in two different periods or by two different source populations. The mitochondrial DNA phylogeographical structuring is not completely concordant with the subspecies distribution. These findings suggest that rock partridges should be managed based on the identification of phylogeographical units.


Population Dynamics of the Sea Slug Plakobranchus ocellatus (Opisthobranch: Sacoglossa: Elysioidea) on a Subtropical Coral Reef off Okinawa-jima Island, Ryukyu Archipelago, Japan.
Daisuke Tanamura and Euichi Hirose (2016) Plakobranchus ocellatus is a sacoglossan sea slug that can retain functional chloroplast from its algal food. This species feed on multiple species of siphonous green algae and can survive several months without food by utilizing retained chloroplasts in its digestive gland (kleptoplasty). While the population dynamics of opisthobranchs are often influenced by the seasonal fluctuation of the abundance of food resources, the fluctuation of food availability would not be a crucial factor to restrict the occurrence of P. ocellatus. We monitored the population density of P. ocellatus for 20 months on a subtropical coral reef where the water temperature fluctuated from 17°C to 32°C, in order to examine whether the population density, distribution pattern of individuals, and size distribution of P. ocellatus are stable or seasonally change. The present results showed that P. ocellatus appeared all year round in the study site, while the population density changed seasonally. The population density decreased in cold (≤ 21°C) and hot (≥ 27°C) periods, and densities in the months of intermediate temperature range (< 21°C, > 25°C) were significantly higher than the densities in other months (Student's t-test, P < 0.0001). Accordingly, population density is probably influenced by water temperature. Morisita's Iδ indicated that the sea slugs were distributed in random patterns (13 months) or clumped patterns (7 months). Our field observations indicated that the sea slugs do not feed in daytime, and probably feed at night. Whereas P. ocellatus individuals of less than 10 mm were rarely recorded in the monitoring area, a decrease of the average body length and increase in population density in April - May suggest active recruitment of small individuals in this period.


Activation of the cholinergic anti-inflammatory pathway ameliorates postoperative ileus in mice.
We previously showed that intestinal inflammation is reduced by electrical stimulation of the efferent vagus nerve, which prevents postoperative ileus in mice. We propose that this cholinergic anti-inflammatory pathway is mediated via alpha7 nicotinic acetylcholine receptors expressed on macrophages. The aim of this study was to evaluate pharmacologic activation of the cholinergic anti-inflammatory pathway in a mouse model for postoperative ileus using the alpha7 nicotinic acetylcholine receptor-agonist AR-R17779. Mice were pretreated with vehicle, nicotine, or AR-R17779 20 minutes before a laparotomy (L) or intestinal manipulation (IM). Twenty-four hours thereafter gastric emptying was determined using scintigraphy and intestinal muscle inflammation was quantified. Nuclear factor-kappaB transcriptional activity and cytokine production was assayed in peritoneal macrophages. Twenty-four hours after surgery IM led to a delayed gastric emptying compared with L (gastric retention: L(saline) 14% +/- 4% vs IM(saline) 38% +/- 10%, P = .04). Pretreatment with AR-R17779 prevented delayed gastric emptying (IM(AR-R17779) 15% +/- 4%, P = .03). IM elicited inflammatory cell recruitment (L(saline) 50 +/- 8 vs IM(saline) 434 +/- 71 cells/mm(2), P = .001) which was reduced by AR-R17779 pretreatment (IM(AR-R17779) 231 +/- 32 cells/mm(2), P = .04). An equimolar dose of nicotine was not tolerated. Subdiaphragmal vagotomy did not affect the anti-inflammatory properties of AR-R17779. In peritoneal macrophages, both nicotinic agonists reduced nuclear factor kappaB transcriptional activity and proinflammatory cytokine production, with nicotine being more effective than AR-R17779. AR-R17779 treatment potently prevents postoperative ileus, whereas toxicity limits nicotine administration to ineffective doses. Our data further imply that nicotinic inhibition of macrophage activation may involve other receptors in addition to alpha7 nicotinic acetylcholine receptor.


Ultrastructural characterization of noradrenergic axons and Beta-adrenergic receptors in the lateral nucleus of the amygdala.
Norepinephrine (NE) is thought to play a key role in fear and anxiety, but its role in amygdala-dependent Pavlovian fear conditioning, a major model for understanding the neural basis of fear, is poorly understood. The lateral nucleus of the amygdala (LA) is a critical brain region for fear learning and regulating the effects of stress on memory. To understand better the cellular mechanisms of NE and its adrenergic receptors in the LA, we used antibodies directed against dopamine beta-hydroxylase (DβH), the synthetic enzyme for NE, or against two different isoforms of the beta-adrenergic receptors (βARs), one that predominately recognizes neurons (βAR 248) and the other astrocytes (βAR 404), to characterize the microenvironments of DβH and βAR. By electron microscopy, most DβH terminals did not make synapses, but when they did, they formed both asymmetric and symmetric synapses. By light microscopy, βARs were present in both neurons and astrocytes. Confocal microscopy revealed that both excitatory and inhibitory neurons express βAR248. By electron microscopy, βAR 248 was present in neuronal cell bodies, dendritic shafts and spines, and some axon terminals and astrocytes. When in dendrites and spines, βAR 248 was frequently concentrated along plasma membranes and at post-synaptic densities of asymmetric (excitatory) synapses. βAR 404 was expressed predominately in astrocytic cell bodies and processes. These astrocytic processes were frequently interposed between unlabeled terminals or ensheathed asymmetric synapses. Our findings provide a morphological basis for understanding ways in which NE may modulate transmission by acting via synaptic or non-synaptic mechanisms in the LA.


Lessons from BWS twins: complex maternal and paternal hypomethylation and a common source of haematopoietic stem cells.
The Beckwith-Wiedemann syndrome (BWS) is a growth disorder for which an increased frequency of monozygotic (MZ) twinning has been reported. With few exceptions, these twins are discordant for BWS and for females. Here, we describe the molecular and phenotypic analysis of 12 BWS twins and a triplet; seven twins are MZ, monochorionic and diamniotic, three twins are MZ, dichorionic and diamniotic and three twins are dizygotic. Twelve twins are female. In the majority of the twin pairs (11 of 13), the defect on chromosome 11p15 was hypomethylation of the paternal allele of DMR2. In 5 of 10 twins, there was additional hypomethylation of imprinted loci; in most cases, the loci affected were maternally methylated, but in two cases, hypomethylation of the paternally methylated DLK1 and H19 DMRs was detected, a novel finding in BWS. In buccal swabs of the MZ twins who share a placenta, the defect was present only in the affected twin; comparable hypomethylation in lymphocytes was detected in both the twins. The level of hypomethylation reached levels below 25%. The exchange of blood cells through vascular connections cannot fully explain the degree of hypomethylation found in the blood cell of the non-affected twin. We propose an additional mechanism through which sharing of aberrant methylation patterns in discordant twins, limited to blood cells, might occur. In a BWS-discordant MZ triplet, an intermediate level of demethylation was found in one of the non-affected sibs; this child showed mild signs of BWS. This finding supports the theory that a methylation error proceeds and possibly triggers the twinning process.


Amygdalin-mediated inhibition of non-small cell lung cancer cell invasion in vitro.
Lung cancer is a common malignant tumor claiming the highest fatality worldwide for a long period of time. Unfortunately, most of the current treatment methods are still based on the characteristics of cancer cells in the primary lesion and the prognosis is often much poorer in patients with metastatic cancers. Amygdalin, a natural product of glycosides and lots of evidence shows that amygdalin can inhibit the proliferation of some kinds of cancer cells. In this study, we first obtained the highly metastatic NSCLC cell lines H1299/M and PA/M and further treated these cells with amygdalin. We found that the in vitro proliferability of H1299/M and PA/M was inhibited, but such inhibition required higher concentration of amygdalin. When lower concentration of amygdalin was used for the experiments, we observed that the in vitro invasive and migration capacities of H1299/M and PA/M were significantly inhibited. These results strongly suggested that amygdalin was likely to have anti-metastatic NSCLC effect. This study offers information of the role of amygdalin that may be useful as a therapeutic target in lung tumors.


Assessing Primary Care Physician's Beliefs and Attitudes of Asthma Exacerbation Treatment and Follow-Up.
The objective of this survey was to assess adult primary care physicians' and pediatricians' perceptions of asthma exacerbation management, including beliefs concerning the discharge of patients from the emergency department (ED) following asthma exacerbations. This was a cross-sectional survey of primary care physicians (PCPs) treating adult or pediatric patients. Surveys were mailed to physicians and included questions on how PCPs define an exacerbation, how they are notified and how they followed-up with their patients who experienced exacerbations. A total of 189 physicians were targeted in this survey, with 124 (65%) returning a completed survey. The majority of physicians agreed that an exacerbation included worsening asthma requiring a course of oral corticosteroids (83%). However, >/=70% of physicians agreed that an exacerbation could also include events which did not require OCS. Overall, 71% of PCPs believed that the majority of their patients' asthma exacerbations were treated in the doctor's office with only 6% believing the majority were treated in the ED. Over 90% of PCPs surveyed said they scheduled a follow-up with their patients "all or most of the time" when notified of an ED visit for an asthma exacerbation. Of the adult PCPs surveyed, 20% said they were never notified when one of their patients received treatment in the hospital because of an asthma exacerbation, whereas only 10% of pediatricians said they were never notified. The majority of PCPs surveyed (79%) indicated that if a controller medication was warranted, the ED staff should initiate treatment at time of discharge. This study showed that healthcare providers may not share a common definition of an asthma exacerbation. In addition, most physicians believe that the majority of exacerbations are treated in their office or at home. Further, most agreed that if a controller medication was warranted, the ED or urgent care staff should initiate treatment.


The Elephant in the Room: Too Much Medicine in Musculoskeletal Practice.
While advances in assessment and management of musculoskeletal conditions have improved care for many people, there have been other, less beneficial developments in the provision of care for people with musculoskeletal pain conditions, one of which is the worrying tendency to provide too much medicine. Too much medicine occurs when the provision of either investigation or intervention (or both) is unjustifiably excessive. Another concern in musculoskeletal health care is medicalizing normality-when a normal human function or condition is labeled as abnormal. In this Viewpoint, the authors argue that medicalizing normality creates health concerns where none exist, while too much medicine involves provision of care where benefits do not outweigh harms, and wastes precious health care resources. The authors (1) list 2 common examples of too much medicine, and 2 examples of medicalizing normality, relevant to physical therapy practice; (2) outline the drivers of too much medicine and medicalizing normality; and (3) make suggestions for change. J Orthop Sports Phys Ther 2020;50(1):1-4. doi:10.2519/jospt.2020.0601.


Too much to know? The cognitive demands of daily knowledge seeking and the buffering role of coworker contact quality.
To get their work done and achieve their daily work-related goals, employees seek knowledge from their coworkers. While the benefits of knowledge seeking have been established in the literature, we have yet to understand the potential downsides of daily knowledge seeking. We adopt a cognitive perspective to carve out the negative effect of daily knowledge seeking, while controlling for its established positive effect via perceived learning. Based on cognitive load theory, we argue that daily knowledge seeking produces intrinsic cognitive load that can hinder daily goal attainment through the experience of knowledge overload and subsequent resource depletion. However, the relational context in which knowledge seekers interact with knowledge sources represents an important contextual boundary condition. Coworker contact quality can mitigate the effect of knowledge seeking on knowledge overload because high coworker contact quality reduces extraneous (i.e., ineffective) and increases germane (i.e., productive) cognitive load that knowledge seekers experience when navigating the social interaction with knowledge sources. Under this condition, cognitive capacity is freed up and knowledge overload is less likely to occur. Based on an experience sampling study in which we collected data across 10 working days from 189 German employees, we found support for our hypotheses. An employee's knowledge seeking had a negative indirect effect on goal attainment via knowledge overload and subsequent resource depletion, however, the downsides of daily knowledge seeking became less pronounced when coworker contact quality increased. We discuss the implications of our findings for research on knowledge seeking and resource exchange behaviors. (PsycInfo Database Record (c) 2022 APA, all rights reserved).


Assessment of prevalence and mortality incidences due to poisoning in a South Indian tertiary care teaching hospital.
WHO reports estimate poisoning as one of the most common causes of increased morbidity and mortality rate world-wide. Various agents such as pesticides, drugs have been used for intentional and accidental poisoning in different countries. In the Indian scenario, pesticides are the most commonly used poisoning agents. To assess the prevalence and mortality incidence rate due to various poisoning agents a retrospective and prospective study conducted over a period of nine months in a tertiary care teaching hospital. Retrospective data of poisoning cases was collected from the medical records section and the prospective data of poisoning cases was collected from the emergency and causality departments. A total of 1045 poisoning related admissions were identified during the period January 2005 to September 2008. Among them, 68.40% of cases were due to intentional poisoning and 31.60% were due to accidental poisoning. Of the poisoning related admissions, 84.4% of patients recovered, whereas in 7.6% of cases condition did not improve. Mortality rate was observed 4%. Intentional poisoning was observed more in male population (60.2%) in the age group of 18-29 years. Accidental poisoning was seen more in children in the age group of 1-3 years. Incidence of overall poisoning cases were high due to pesticides (39.5%) followed by medicines (26.1%), household products (22.1%), environmental poisoning (12.1%) and heavy metals (0.2%). It was observed that availability of centralised poison information centre and treatment protocols will improvise poison management practices in tertiary care hospitals by the clinicians.


Analysis of the MTHFD1 promoter and risk of neural tube defects.
Genetic variants in MTHFD1 (5,10-methylenetetrahydrofolate dehydrogenase/5,10-methenyltetrahydrofolate cyclohydrolase/ 10-formyltetrahydrofolate synthetase), an important folate metabolic enzyme, are associated with a number of common diseases, including neural tube defects (NTDs). This study investigates the promoter of the human MTHFD1 gene in a bid to understand how this gene is controlled and regulated. Following a combination of in silico and molecular approaches, we report that MTHFD1 expression is controlled by a TATA-less, Initiator-less promoter and transcription is initiated at multiple start sites over a 126 bp region. We confirmed the presence of three database polymorphisms (dbSNP) by direct sequencing of the upstream region (rs1076991 C > T, rs8010584 G > A, rs4243628 G > T), with a fourth (dbSNP rs746488 A > T) not found to be polymorphic in our population and no novel polymorphisms identified. We demonstrate that a common SNP rs1076991 C > T within the window of transcriptional initiation exerts a significant effect on promoter activity in vitro. We investigated this SNP as a potential risk factor for NTDs in a large homogenous Irish population and determined that it is not an independent risk factor, but, it does increase both case (chi (2) = 11.06, P = 0.001) and maternal (chi (2) = 6.68, P = 0.01) risk when allele frequencies were analysed in combination with the previously identified disease-associated p.R653Q (c.1958 G > A; dbSNP rs2236225) polymorphism. These results provide the first insight into how MTHFD1 is regulated and further emphasise its importance during embryonic development.


Comparative Genomic Insights into Ecophysiology of Neutrophilic, Microaerophilic Iron Oxidizing Bacteria.
Neutrophilic microaerophilic iron-oxidizing bacteria (FeOB) are thought to play a significant role in cycling of carbon, iron and associated elements in both freshwater and marine iron-rich environments. However, the roles of the neutrophilic microaerophilic FeOB are still poorly understood due largely to the difficulty of cultivation and lack of functional gene markers. Here, we analyze the genomes of two freshwater neutrophilic microaerophilic stalk-forming FeOB, Ferriphaselus amnicola OYT1 and Ferriphaselus strain R-1. Phylogenetic analyses confirm that these are distinct species within Betaproteobacteria; we describe strain R-1 and propose the name F. globulitus. We compare the genomes to those of two freshwater Betaproteobacterial and three marine Zetaproteobacterial FeOB isolates in order to look for mechanisms common to all FeOB, or just stalk-forming FeOB. The OYT1 and R-1 genomes both contain homologs to cyc2, which encodes a protein that has been shown to oxidize Fe in the acidophilic FeOB, Acidithiobacillus ferrooxidans. This c-type cytochrome common to all seven microaerophilic FeOB isolates, strengthening the case for its common utility in the Fe oxidation pathway. In contrast, the OYT1 and R-1 genomes lack mto genes found in other freshwater FeOB. OYT1 and R-1 both have genes that suggest they can oxidize sulfur species. Both have the genes necessary to fix carbon by the Calvin-Benson-Basshom pathway, while only OYT1 has the genes necessary to fix nitrogen. The stalk-forming FeOB share xag genes that may help form the polysaccharide structure of stalks. Both OYT1 and R-1 make a novel biomineralization structure, short rod-shaped Fe oxyhydroxides much smaller than their stalks; these oxides are constantly shed, and may be a vector for C, P, and metal transport to downstream environments. Our results show that while different FeOB are adapted to particular niches, freshwater and marine FeOB likely share common mechanisms for Fe oxidation electron transport and biomineralization pathways.


Genetic analysis and QTL mapping for multiple biotic stress resistance in cassava.
In Sub-Saharan Africa cassava (Manihot esculenta Crantz) is one of the most important food crops where more than 40% of the population relies on it as their staple carbohydrate source. Biotic constraints such as viral diseases, mainly Cassava Mosaic Disease (CMD) and Cassava Brown Streak Disease (CBSD), and arthropod pests, particularly Cassava Green Mite (CGM), are major constraints to the realization of cassava's full production potential in Africa. To address these problems, we aimed to map the quantitative trait loci (QTL) associated with resistance to CBSD foliar and root necrosis symptoms, foliar CMD and CGM symptoms in a full-sib mapping population derived from the genotypes AR40-6 and Albert. A high-density linkage map was constructed with 2,125 SNP markers using a genotyping-by-sequencing approach. For phenotyping, clonal evaluation trials were conducted with 120 F1 individuals for two consecutive field seasons using an alpha-lattice design at Chambezi and Naliendele, Tanzania. Previously identified QTL for resistance to CBSD foliar symptoms were corroborated, and a new putative QTL for CBSD root necrosis identified (qCBSDRNc14AR) from AR40-6. Two QTL were identified within the region of the previously recognized CMD2 locus from this population in which both parents are thought to possess the CMD2 locus. Interestingly, a minor but consistent QTL, qCGM18AR, for CGM resistance at 3 months after planting stage was also detected and co-localized with a previously identified SSR marker, NS346, linked with CGM resistance. Markers underlying these QTL may be used to increase efficiencies in cassava breeding programs.


Predictors of treatment receipt among adults with a drug use disorder.
This study used data from the 2000 and 2001 National Household Surveys on Drug Abuse to examine factors that contribute to the receipt of specialty substance abuse treatment, which is defined as treatment in rehabilitation facilities, hospitals, or mental health centers designed to help stop or reduce drug use. The population examined was a nationally representative sample of 3291 adults aged 18 or older with a drug use disorder in the past 12 months. Data were collected by computer-assisted interviews using a combination of computer-assisted personal interviews conducted by the interviewer and audio computer-assisted self-interviewing guided by the computer and respondent. Using descriptive analyses and multivariate logistic regression models, this study compared sociodemographic, substance abuse, and psychosocial characteristics of those receiving treatment with those not receiving treatment; it also examined the factors that influenced treatment receipt while controlling for potential confounders. Characteristics significantly contributing to treatment receipt among adults with a drug use disorder included the following: a woman without social support; a high school graduate with no college education; those receiving insurance through Medicaid or a state Children's Health Insurance Program; those on probation, parole, or supervised release in the past year; a daily smoker of cigarettes; those meeting at least three criteria for drug dependence; those having past year dependence on or abuse of alcohol; and those receiving any mental health treatment or counseling in the past year. Adults associated with the criminal justice system had a different pattern of treatment predictors from those who were not involved with the criminal justice system.


Topography of age-related changes in sleep spindles.
Aging induces multiple changes to sleep spindles, which may hinder their alleged functional role in memory and sleep protection mechanisms. Brain aging in specific cortical regions could affect the neural networks underlying spindle generation, yet the topography of these age-related changes is currently unknown. In the present study, we analyzed spindle characteristics in 114 healthy volunteers aged between 20 and 73 years over 5 anteroposterior electroencephalography scalp derivations. Spindle density, amplitude, and duration were higher in young subjects than in middle-aged and elderly subjects in all derivations, but the topography of age effects differed drastically. Age-related decline in density and amplitude was more prominent in anterior derivations, whereas duration showed a posterior prominence. Age groups did not differ in all-night spindle frequency for any derivation. These results show that age-related changes in sleep spindles follow distinct topographical patterns that are specific to each spindle characteristic. This topographical specificity may provide a useful biomarker to localize age-sensitive changes in underlying neural systems during normal and pathological aging.


Neural mechanisms of discourse comprehension: a human lesion study.
Discourse comprehension is a hallmark of human social behaviour and refers to the act of interpreting a written or spoken message by constructing mental representations that integrate incoming language with prior knowledge and experience. Here, we report a human lesion study (n = 145) that investigates the neural mechanisms underlying discourse comprehension (measured by the Discourse Comprehension Test) and systematically examine its relation to a broad range of psychological factors, including psychometric intelligence (measured by the Wechsler Adult Intelligence Scale), emotional intelligence (measured by the Mayer, Salovey, Caruso Emotional Intelligence Test), and personality traits (measured by the Neuroticism-Extraversion-Openness Personality Inventory). Scores obtained from these factors were submitted to voxel-based lesion-symptom mapping to elucidate their neural substrates. Stepwise regression analyses revealed that working memory and extraversion reliably predict individual differences in discourse comprehension: higher working memory scores and lower extraversion levels predict better discourse comprehension performance. Lesion mapping results indicated that these convergent variables depend on a shared network of frontal and parietal regions, including white matter association tracts that bind these areas into a coordinated system. The observed findings motivate an integrative framework for understanding the neural foundations of discourse comprehension, suggesting that core elements of discourse processing emerge from a distributed network of brain regions that support specific competencies for executive and social function.


Property profiling of biosimilar mucus in a novel mucus-containing in vitro model for assessment of intestinal drug absorption.
Oral delivery of drugs, including peptide and protein therapeutics, can be impeded by the presence of the mucus surface-lining the intestinal epithelium. The aim of the present project was to design and characterize biosimilar mucus compatible with Caco-2 cell monolayers cultured in vitro to establish a more representative in vitro model for the intestinal mucosa. The rheological profile of a biosimilar mucus mixture composed of purified gastric mucin, lipids and protein in buffer was optimized by supplementing with an anionic polymer to display viscoelastic properties and a microstructure comparable to freshly isolated porcine intestinal mucus (PIM). Further, this multicomponent biosimilar mucus mixture was optimized with regard to the lipid content in order to obtain cellular compatibility with well-differentiated Caco-2 cell monolayers. In contrast, PIM was found to severely disrupt the Caco-2 cell monolayer. When combined with the Caco-2 cell monolayers, the final biosimilar mucus was found to significantly affect the permeability profiles for hydrophobic and hydrophilic small and large model drug compounds in different ways. In conclusion, the present study describes an improvement of the biorelevance of the Caco-2 cell culture model by application of mucus, resulting in an in vitro model of oral mucosa suitable for future assessment of innovative drug delivery approaches.


Randomized comparison of primary stenting and provisional balloon angioplasty guided by flow velocity measurement. Doppler Endpoints Balloon Angioplasty Trial Europe (DEBATE) II Study Group.
Coronary stenting improves outcomes compared with balloon angioplasty, but it is costly and may have other disadvantages. Limiting stent use to patients with a suboptimal result after angioplasty (provisional angioplasty) may be as effective and less expensive. To analyze the cost-effectiveness of provisional angioplasty, patients scheduled for single-vessel angioplasty were first randomized to receive primary stenting (97 patients) or balloon angioplasty guided by Doppler flow velocity and angiography (523 patients). Patients in the latter group were further randomized after optimization to either additional stenting or termination of the procedure to further investigate what is "optimal." An optimal result was defined as a flow reserve >2.5 and a diameter stenosis <36%. Bailout stenting was needed in 129 patients (25%) who were randomized to balloon angioplasty, and an optimal result was obtained in 184 of the 523 patients (35%). There was no significant difference in event-free survival at 1 year between primary stenting (86.6%) and provisional angioplasty (85.6%). Costs after 1 year were significantly higher for provisional angioplasty (EUR 6573 versus EUR 5885; P:=0.014). Results after the second randomization showed that stenting was also more effective after optimal balloon angioplasty (1-year event free survival, 93.5% versus 84.1%; P:=0. 066). After 1 year of follow-up, provisional angioplasty was more expensive and without clinical benefit. The beneficial value of stenting is not limited to patients with a suboptimal result after balloon angioplasty.


Regional brain volume abnormalities and long-term cognitive outcome in preterm infants.
Preterm infants have a high prevalence of long-term cognitive and behavioral disturbances. However, it is not known whether the stresses associated with premature birth disrupt regionally specific brain maturation or whether abnormalities in brain structure contribute to cognitive deficits. To determine whether regional brain volumes differ between term and preterm children and to examine the association of regional brain volumes in prematurely born children with long-term cognitive outcomes. Case-control study conducted in 1998 and 1999 at 2 US university medical schools. A consecutive sample of 25 eight-year-old preterm children recruited from a longitudinal follow-up study of preterm infants and 39 term control children who were recruited from the community and who were comparable with the preterm children in age, sex, maternal education, and minority status. Volumes of cortical subdivisions, ventricular system, cerebellum, basal ganglia, corpus callosum, amygdala, and hippocampus, derived from structural magnetic resonance imaging scans and compared between preterm and term children; correlations of regional brain volumes with cognitive measures (at age 8 years) and perinatal variables among preterm children. Regional cortical volumes were significantly smaller in the preterm children, most prominently in sensorimotor regions (difference: left, 14.6%; right, 14.3% [P<.001 for both]) but also in premotor (left, 11.2%; right, 12.6% [P<.001 for both]), midtemporal (left, 7.4% [P =.01]; right, 10.2% [P<.001]), parieto-occipital (left, 7.9% [P =.01]; right, 7.4% [P =.005]), and subgenual (left, 8.9% [P =.03]; right, 11.7% [P =.01]) cortices. Preterm children's brain volumes were significantly larger (by 105. 7%-271.6%) in the occipital and temporal horns of the ventricles (P<. 001 for all) and smaller in the cerebellum (6.7%; P =.02), basal ganglia (11.4%-13.8%; P</=.005), amygdala (left, 20.2% [P =.001]; right, 30.0% [P<.001]), hippocampus (left, 16.0% [P =.001]; right, 12.0% [P =.007]), and corpus callosum (13.1%-35.2%; P</=.01 for all). Volumes of sensorimotor and midtemporal cortices were associated positively with full-scale, verbal, and performance IQ scores (P<.01 for all). Our data indicate that preterm birth is associated with regionally specific, long-term reductions in brain volume and that morphological abnormalities are, in turn, associated with poorer cognitive outcome. JAMA. 2000;284:1939-1947.


Emergent exploration via novelty management.
When encountering novel environments, animals perform complex yet structured exploratory behaviors. Despite their typical structuring, the principles underlying exploratory patterns are still not sufficiently understood. Here we analyzed exploratory behavioral data from two modalities: whisking and locomotion in rats and mice. We found that these rodents maximized novelty signal-to-noise ratio during each exploration episode, where novelty is defined as the accumulated information gain. We further found that these rodents maximized novelty during outbound exploration, used novelty-triggered withdrawal-like retreat behavior, and explored the environment in a novelty-descending sequence. We applied a hierarchical curiosity model, which incorporates these principles, to both modalities. We show that the model captures the major components of exploratory behavior in multiple timescales: single excursions, exploratory episodes, and developmental timeline. The model predicted that novelty is managed across exploratory modalities. Using a novel experimental setup in which mice encountered a novel object for the first time in their life, we tested and validated this prediction. Further predictions, related to the development of brain circuitry, are described. This study demonstrates that rodents select exploratory actions according to a novelty management framework and suggests a plausible mechanism by which mammalian exploration primitives can be learned during development and integrated in adult exploration of complex environments.


Experiences of Self-Monitoring in Self-Directed Weight Loss and Weight Loss Maintenance: Systematic Review of Qualitative Studies.
The experience and role of self-monitoring in self-directed weight loss attempts may be distinctly different from that within formal interventions, and has yet to be fully explored. We systematically reviewed qualitative studies to examine experiences of self-monitoring as an aid to self-directed weight loss. Thematic synthesis was used to construct descriptive and analytical themes from the available data. In all, 22 studies (681 participants) were included, in which the uses of self-monitoring ranged from an aid to increase adherence to a tool for facilitating analysis. Self-monitoring also influenced and was influenced by self-perception and emotions. Feelings of shame were linked with abandonment of efforts. Findings highlight the centrality of interpretation of self-monitored data, the implications this interpretation has on sense of self, and the impact of broader discourses. Explicitly framing self-monitoring as a positive tool with which to aid analysis may encourage helpful use of this technique.


Designing an anion-functionalized fluorescent ionic liquid as an efficient and reversible turn-off sensor for detecting SO2.
A novel anion-functionalized fluorescent ionic liquid was designed and prepared, which was capable of capturing sulphur dioxide with high capacity and could also be used as a good colorimetric and fluorescent SO2 sensor. Compared to conventional fluorescent sensors, this fluorescent ionic liquid did not undergo aggregation-caused quenching or aggregation-induced emission, and the fluorescence was quenched when exposed to SO2, and the fluorescence would quench when exposed to SO2. The experimental absorption, spectroscopic investigation, and quantum chemical calculations indicated that the quenching of the fluorescence originated from SO2 physical absorption, not chemical absorption. Furthermore, this fluorescent ionic liquid exhibited high selectivity, good quantification, and excellent reversibility for SO2 detection, and showed potential for an excellent liquid sensor.


The qualities of an effective mentor from the student nurse's perspective: findings from a longitudinal qualitative study.
Parker and Carlisle (Journal of Advanced Nursing 24, 771-778) argue that there is a scarcity of empirical research focusing on issues such as supernumerary status and mentorship in Project 2000 courses from the students' perspective. This paper presents the findings of a longitudinal cohort study using Grounded Theory to discover the effect(s) of mentorship on student nurses following the introduction of the 1992 programme of education leading to a Diploma of Higher Education in Nursing and registration with the United Kingdom Central Council (UKCC). The cohort consisted of 10 students from a large Scottish College of Nursing & Midwifery who were interviewed on five occasions during the three years of their course. Students also kept a diary to record their thoughts and experiences regarding mentorship during their practice placements. In addition, a further seven students volunteered to participate by diary only. Data were analysed with the aid of NUD.IST and subjected to the constant comparative method of analysis. Findings indicate that Diploma students quickly lose their idealistic view of their mentor and over time develop an insight into the qualities they perceive are required of an effective mentor. Students quickly become aware of the importance of choosing good role models and learning their own mentor's likes and dislikes as they realize this impinges on the outcome of their assessment. As students move into their Branch programme, a gradual distancing from their mentor is evident. This coincides with a development in their confidence, skills and a holistic perspective of care.


Are high lags of accommodation in myopic children due to motor deficits?
Children with a progressing myopia exhibit an abnormal pattern of high accommodative lags coupled with high accommodative convergence (AC/A) and high accommodative adaptation. This is not predicted by the current models of accommodation and vergence. Reduced accommodative plant gain and reduced sensitivity to blur have been suggested as potential causes for this abnormal behavior. These etiologies were tested by altering parameters (sensory, controller and plant gains) in the Simulink model of accommodation. Predictions were then compared to the static and dynamic blur accommodation (BA) measures taken using a Badal optical system on 12 children (6 emmetropes and 6 myopes, 8-13years) and 6 adults (20-35years). Other critical parameters such as CA/C, AC/A, and accommodative adaptation were also measured. Usable BA responses were classified as either typical or atypical. Typical accommodation data confirmed the abnormal pattern of myopia along with an unchanged CA/C. Main sequence relationship remained invariant between myopic and nonmyopic children. An overall reduction was noted in the response dynamics such as peak velocity and acceleration with age. Neither a reduced plant gain nor reduced blur sensitivity could predict the abnormal accommodative behavior. A model adjustment reflecting a reduced accommodative sensory gain (ASG) coupled with an increased AC cross-link gain and reduced vergence adaptive gain does predict the empirical findings. Empirical measures also showed a greater frequency of errors in accommodative response generation (atypical responses) in both myopic and control children compared to adults.


What gets published: the characteristics of quality improvement research articles from low- and middle-income countries.
Reports of quality improvement (QI) research from low- and middle-income countries (LMICs) remain sparse in the scientific literature. The authors reviewed the published literature to describe the characteristics of such reports. The authors conducted a systematic search for QI research articles from LMICs catalogued in the PubMed databases prior to December 2011, complemented by recommendations from experts in the field. Articles were categorised based on bibliometric and research characteristics. Twenty papers were randomly selected for narrative analysis regarding strategies used to present the methods and results of interventions. Seventy-six articles met the inclusion criteria. Publication rate accelerated over time, particularly among observational studies. Most studies did not use a concurrent control group; pre-/post-study designs were most common overall. Four papers were published in top-tier journals, 17 in journals at the top of their specialty and 20 in quality-specific journals. Among the papers selected for narrative analysis, four distinct components were observed in most: a description of the problem state, a description of the improvement processes and tools, a separate description of the interventions tested and a description of the evaluation methods. The small number of articles identified by this review suggests that publication of QI research from LMICs remains challenging. However, recent increases in publication rates, especially among observational studies, may attest to greater interest in the topic among scientific audiences. Though the authors are not able to compare this sample with unpublished papers, the four components observed by them in the narrative analysis seem to strengthen QI research reports.


Investigating tendon mineralisation in the avian hindlimb: a model for tendon ageing, injury and disease.
Mineralisation of the tendon tissue has been described in various models of injury, ageing and disease. Often resulting in painful and debilitating conditions, the processes underlying this mechanism are poorly understood. To elucidate the progression from healthy tendon to mineralised tendon, an appropriate model is required. In this study, we describe the spontaneous and non-pathological ossification and calcification of tendons of the hindlimb of the domestic chicken (Gallus gallus domesticus). The appearance of the ossified avian tendon has been described previously, although there have been no studies investigating the developmental processes and underlying mechanisms leading to the ossified avian tendon. The tissue and cells from three tendons - the ossifying extensor and flexor digitorum longus tendons and the non-ossifying Achilles tendon - were analysed for markers of ageing and mineralisation using histology, immunohistochemistry, cytochemistry and molecular analysis. Histologically, the adult tissue showed a loss of healthy tendon crimp morphology as well as markers of calcium deposits and mineralisation. The tissue showed a lowered expression of collagens inherent to the tendon extracellular matrix and presented proteins expressed by bone. The cells from the ossified tendons showed a chondrogenic and osteogenic phenotype as well as tenogenic phenotype and expressed the same markers of ossification and calcification as the tissue. A molecular analysis of the gene expression of the cells confirmed these results. Tendon ossification within the ossified avian tendon seems to be the result of an endochondral process driven by its cells, although the roles of the different cell populations have yet to be elucidated. Understanding the role of the tenocyte within this tissue and the process behind tendon ossification may help us prevent or treat ossification that occurs in injured, ageing or diseased tendon.


Genetic studies of membrane excitability in Drosophila: lethal interaction between two temperature-sensitive paralytic mutations.
Two mutants of Drosophila melanogaster, parats1 (1-53.9) and napts (2-56.2) both display similar temperature-sensitive paralysis associated with blockage in the conduction of nerve action potentials, suggesting that the two gene products have a similar function. This idea is supported by the observation that the double mutant is unconditionally lethal. Genetic analysis of this synergistic interaction has revealed the following: it specifically involves the para and nap loci; all para alleles interact with napts, but the strength of the interaction varies in an allele-dependent fashion; lethality of the double mutant occurs during the first larval instar with parats1 but differs with other para alleles; hypodosage of para+ causes lethality in a napts background. These results together with previous electrophysiological, behavioral and pharmacological studies of these mutants suggest that both para and nap affect sodium channels and possibly encode different subunits.


Gene-environment interaction demonstrates the vulnerability of the embryonic heart.
Mammalian embryos develop in a low oxygen environment. The transcription factor hypoxia inducible factor 1a (HIF1α) is a key element in the cellular response to hypoxia. Complete deletion of Hif1α from the mouse conceptus causes extensive placental, vascular and heart defects, resulting in embryonic lethality. However the precise role of Hif1α in each of these organ systems remains unknown. To further investigate, we conditionally-deleted Hif1α from mesoderm, vasculature and heart individually. Surprisingly, deletion from these tissues did not recapitulate the same severe heart phenotype or embryonic lethality. Placental insufficiency, such as occurs in the complete Hif1α null, results in elevated cellular hypoxia in mouse embryos. We hypothesized that subjecting the Hif1α conditional null embryos to increased hypoxic stress might exacerbate the effects of tissue-specific Hif1α deletion. We tested this hypothesis using a model system mimicking placental insufficiency. We found that the majority of embryos lacking Hif1α in the heart died when exposed to non-physiological hypoxia. This was a heart-specific phenomenon, as HIF1α protein accumulated predominantly in the myocardium of hypoxia-stressed embryos. Our study demonstrates the vulnerability of the heart to lowered oxygen levels, and that under such conditions of non-physiological hypoxia the embryo absolutely requires Hif1α to continue normal development. Importantly, these findings extend our understanding of the roles of Hif1α in cardiovascular development.


Temporal trends and predictors of perioperative chemotherapy use in elderly patients with resected nonsmall cell lung cancer.
The authors assessed patterns of perioperative chemotherapy use in elderly patients with resected stage I, II, or IIIA nonsmall cell lung cancer (NSCLC) from 1992 to 2002. By using data from the Surveillance, Epidemiology, and End Results Program, 11,807 patients were identified who had resected stage I, II, or IIIA NSCLC between 1992 and 2002 and survived >or=120 days beyond diagnosis. The rate of perioperative chemotherapy use was measured by calendar year, and the association between clinical/demographic characteristics and the receipt of chemotherapy was examined by using logistic regression. In total, 957 patients with stage I, II, or IIIA NSCLC (8.1% of the study population) received perioperative chemotherapy. The proportion of patients receiving chemotherapy for stage I NSCLC changed little during the study period. Of 3230 patients with stage II and IIIA NSCLC, 609 patients (18.9%) received chemotherapy, 423 patients (13%) received chemotherapy combined with radiation. 452 patients (15.6%) received adjuvant chemotherapy, and 66 patients (2.3%) received neoadjuvant chemotherapy. The use of chemotherapy increased significantly among patients who were diagnosed after 1994 relative to patients who were diagnosed in 1992 after controlling for sociodemographic and treatment characteristics (P< .001). There was significantly increased use of new-generation chemotherapy agents, such as carboplatin and taxanes (P< .001). The proportion of patients receiving combined-modality therapy also increased significant (P< .001). Younger age, being married, having advanced-stage tumor or adenocarcinoma, having a later diagnosis year, receiving radiation, and seeing an oncologist were predictors for the receipt of chemotherapy (P< .001). A substantial proportion of Medicare beneficiaries with NSCLC received perioperative chemotherapy. Specifically designed prospective trials that focus on older patients are needed.


Thermodynamic modeling of chromium: strong and weak magnetic coupling.
As chromium is a decisive ingredient for stainless steels, a reliable understanding of its thermodynamic properties is indispensable. Parameter-free first-principles methods have nowadays evolved to a state allowing such thermodynamic predictions. For materials such as Cr, however, the inclusion of magnetic entropy and higher order contributions such as anharmonic entropy is still a formidable task. Employing state-of-the-art ab initio molecular dynamics simulations and statistical concepts, we compute a set of thermodynamic properties based on quasiharmonic, anharmonic, electronic and magnetic free energy contributions from first principles. The magnetic contribution is modeled by an effective nearest-neighbor Heisenberg model, which itself is solved numerically exactly by means of a quantum Monte Carlo method. We investigate two different scenarios: a weak magnetic coupling scenario for Cr, as usually presumed in empirical thermodynamic models, turns out to be in clear disagreement with experimental observations. We show that instead a mixed Hamiltonian including weak and strong magnetic coupling provides a consistent picture with good agreement to experimental thermodynamic data.


Synergistic binding of transcription factors to cell-specific enhancers programs motor neuron identity.
Efficient transcriptional programming promises to open new frontiers in regenerative medicine. However, mechanisms by which programming factors transform cell fate are unknown, preventing more rational selection of factors to generate desirable cell types. Three transcription factors, Ngn2, Isl1 and Lhx3, were sufficient to program rapidly and efficiently spinal motor neuron identity when expressed in differentiating mouse embryonic stem cells. Replacement of Lhx3 by Phox2a led to specification of cranial, rather than spinal, motor neurons. Chromatin immunoprecipitation-sequencing analysis of Isl1, Lhx3 and Phox2a binding sites revealed that the two cell fates were programmed by the recruitment of Isl1-Lhx3 and Isl1-Phox2a complexes to distinct genomic locations characterized by a unique grammar of homeodomain binding motifs. Our findings suggest that synergistic interactions among transcription factors determine the specificity of their recruitment to cell type-specific binding sites and illustrate how a single transcription factor can be repurposed to program different cell types.


Cell Membrane Composition Drives Selectivity and Toxicity of Designed Cyclic Helix-Loop-Helix Peptides with Cell Penetrating and Tumor Suppressor Properties.
The tumor suppressor protein p53 is inactive in a large number of cancers, including some forms of sarcoma, breast cancer, and leukemia, due to overexpression of its intrinsic inhibitors MDM2 and MDMX. Reactivation of p53 tumor suppressor activity, via disruption of interactions between MDM2/X and p53 in the cytosol, is a promising strategy to treat cancer. Peptides able to bind MDM2 and/or MDMX were shown to prevent MDM2/X:p53 interactions, but most possess low cell penetrability, low stability, and/or high toxicity to healthy cells. Recently, the designed peptide cHLH-p53-R was reported to possess high affinity for MDM2, resistance toward proteases, cell-penetrating properties, and toxicity toward cancer cells. This peptide uses a stable cyclic helix-loop-helix (cHLH) scaffold, which includes two helices connected with a Gly loop and cyclized to improve stability. In the current study, we were interested in examining the cell selectivity of cHLH-p53-R, its cellular internalization, and ability to reactivate the p53 pathway. We designed analogues of cHLH-p53-R and employed biochemical and biophysical methodologies using in vitro model membranes and cell-based assays to compare their structure, activity, and mode-of-action. Our studies show that cHLH is an excellent scaffold to stabilize and constrain p53-mimetic peptides with helical conformation, and reveal that anticancer properties of cHLH-p53-R are mediated by its ability to selectively target, cross, and disrupt cancer cell membranes, and not by activation of the p53 pathway. These findings highlight the importance of examining the mode-of-action of designed peptides to fully exploit their potential to develop targeted therapies.


Deficient maturation of aspects of attention and executive functions in early onset schizophrenia.
The few existing long-term, neuropsychological follow-up studies of early onset schizophrenia (EOS) patients have reported relative stability in some cognitive functions but abnormal developmental trajectories in verbal memory, set shifting, aspects of attention, and speed of information processing throughout late adolescence into early adulthood. The current 5-year follow-up study compared the development of specific cognitive functions in EOS patients (N = 17) from the time of first-episode to chronic phase with that of healthy controls (N = 38) and secondarily to patients with other early onset, non-organic, non-affective psychoses (EOP) (N = 11). Speed of processing of executive functions, set shifting, and attention improved significantly in the healthy controls and reflected continuous functional maturation during late adolescence and early adulthood. The developmental progression of attention and set shifting but not speed of processing of executive functions was significantly subnormal in EOS patients. Other specific cognitive functions that had attained functional maturity in the healthy controls before or around the time of the baseline assessment showed normal development in EOS patients during the follow-up period, indicating stable cognitive deficits. These results suggest post-onset developmental deficits in two out of the three aspects of attention and executive functions that have protracted maturational trajectories and that overlap the age of onset of EOS. No significant difference in the development of any specific cognitive function was found between the EOS and EOP group.


Therapy of adrenal insufficiency: an update.
Adrenal insufficiency may be caused by the destruction or altered function of the adrenal gland with a primary deficit in cortisol secretion (primary adrenal insufficiency) or by hypothalamic-pituitary pathologies determining a deficit of ACTH (secondary adrenal insufficiency). The clinical picture is determined by the glucocorticoid deficit, which may in some conditions be accompanied by a deficit of mineralcorticoids and adrenal androgens. The substitutive treatment is aimed at reducing the signs and symptoms of the disease as well as at preventing the development of an addisonian crisis, a clinical emergency characterized by hypovolemic shock. The oral substitutive treatment should attempt at mimicking the normal circadian profile of cortisol secretion, by using the lower possible doses able to guarantee an adequate quality of life to patients. The currently available hydrocortisone or cortisone acetate preparations do not allow an accurate reproduction of the physiological secretion pattern of cortisol. A novel dual-release formulation of hydrocortisone, recently approved by EMEA, represents an advancement in the optimization of the clinical management of patients with adrenal insufficiency. Future clinical trials of immunomodulation or immunoprevention will test the possibility to delay (or prevent) the autoimmune destruction of the adrenal gland in autoimmune Addison's disease.


RNA polymerase III catalysed transcription can be regulated in Saccharomyces cerevisiae by the bacterial tetracycline repressor-operator system.
We have investigated whether the RNA polymerase III-driven transcription of eukaryotic tRNA genes can be regulated by the prokaryotic tetracycline operator-repressor system. The bacterial tet operator (tetO) was inserted at two different positions (-7 and -46) upstream of a tRNA(Glu) (amber) suppressor gene. Both constructs are transcribed in Saccharomyces cerevisiae and yield functional tRNAs as scored by suppression of an amber nonsense mutation in the met8-1 allele. Controlled expression of Tet repressor was achieved by fusing the bacterial tetR gene to the yeast gal1 promoter. This leads to expression of Tet repressor in yeast on galactose--but not on glucose--containing media. Regulation of the su-tRNA gene with the tetO fragment inserted at position -7 has been demonstrated. Under conditions which allow tetR expression, cells exhibit a met- phenotype. This methionine auxotrophy can be conditionally reverted to prototrophy by adding tetracycline. However, a su-tRNA gene with the tetO fragment inserted at position -46 cannot be repressed. Our results demonstrate clearly that the bacterial repressor protein binds to its operator in the yeast genome. Formation of this complex in the vicinity of the pol III transcription initiation site reduces the level of su-tRNA at least 50-fold as concluded from quantitative primer extension analyses. This indicates for the first time that class III gene expression can be regulated by a DNA binding protein with its target site in the 5'-flanking region and that a prokaryotic repressor can confer regulation of a suitably engineered tRNA gene.


Rheological properties, biocompatibility and in vivo performance of new hydrogel-based bone fillers.
Three different heterologous substitutes for bone regeneration, manufactured with equine-derived cortical powder (CP), cancellous chips (CC) and demineralized bone matrix granules (DBM), were compared in in vitro and in vivo settings. We tested: a commercially available bone paste (Osteoplant-Activagen™, consisting of aqueous collagenous carrier, CP, DBM; named A); a second-generation injectable paste (20 kDa polyethylene glycol/hydroxypropyl-methyl cellulose-based hydrogel, CP, DBM; B); a pre-formed bone filler (400 kDa polyethylene oxide/hydroxypropyl-methyl cellulose-based hydrogel, CP, CC, DBM; C). Vitamin C acted as a visco-modulator during C and B β-rays sterilization, modifying graft injectability. For each filler, we examined dissolution in culture medium, gene expression of the substitute-exposed osteogenically-induced human bone marrow stromal cells (hBMSC), and performance in a rabbit bone defect model. A dissolved after 1 h, while fragmentation of B peaked after 8 h. C remained unaltered for 2 days, but affected the microenvironmental pH, slowing the proliferation of exposed cells. B-exposed hBMSC overexpressed bone sialoprotein, osteocalcin and RUNX2. For all fillers histological results evidenced bridged lesion margins, marrow replenishment and bone-remodeling. However, B-treated lesions displayed a metachromatic type II collagen-rich matrix with prehypertrophic-like cells, matching the in vitro expression of cartilage-specific markers, and suggesting a possible application of B/C double-layer monolithic osteochondral plugs for full-thickness articular defects.


Improved visualization of outer retinal morphology with aberration cancelling reflective optical design for adaptive optics - optical coherence tomography.
We present an aberration cancelling optical design for a reflective adaptive optics - optical coherence tomography (AO-OCT) retinal imaging system. The optical performance of this instrument is compared to our previous multimodal AO-OCT/AO-SLO retinal imaging system. The feasibility of new instrumentation for improved visualization of microscopic retinal structures is discussed. Examples of images acquired with this new AO-OCT instrument are presented.


Influence of high hydrostatic pressure on beat length in highly birefringent single-mode bow tie fibers.
New experimental studies on the effects of high hydrostatic pressure on highly birefringent optical fibers are presented. Using Rayleigh scattering, the effect of beat length modification in highly birefringent singlemode bow tie fibers under high pressure up to 100 MPa was investigated. The results indicate that the beat length decreases with pressure, with the mean coefficient of 1/L(B0)|dL(B)/dp| = 0.15%/MPa. A similar decrease of beat length was observed when uniaxial longitudinal stress was applied to the bow tie fiber. A more developed semiphenomenological interpretation of these experimental results is also presented in the paper.


GryphSens: A Smartphone-Based Portable Diagnostic Reader for the Rapid Detection of Progesterone in Milk.
Enzyme-linked immunosorbent assay (ELISA) is a popular assay technique for the detection and quantification of various biological substances due its high sensitivity and specificity. More often, it requires large and expensive laboratory instruments, which makes it difficult to conduct when the tests must be performed quickly at the point-of-care (POC). To increase portability and ease of use, we propose a portable diagnostic system based on a Raspberry Pi imaging sensor for the rapid detection of progesterone in milk samples. We designed, assembled, and tested a standalone portable diagnostic reader and validated it for progesterone detection against a standard ELISA assay using a commercial plate reader. The portable POC device yielded consistent results, regardless of differences in the cameras and flashlights between various smartphone devices. An Android application was built to provide front-end access to users, control the diagnostic reader, and display and store the progesterone measurement on the smartphone. The diagnostic reader takes images of the samples, reads the pixel values, processes the results, and presents the results on the handheld device. The proposed POC reader can perform to superior levels of performance as a plate reader, while adding the desirable qualities of portability and ease of use.