Differential effects of exposure to low-light or high-light open-field on anxiety-related behaviors: relationship to c-Fos expression in serotonergic and non-serotonergic neurons in the dorsal raphe nucleus.
Serotonergic systems arising from the mid-rostrocaudal and caudal dorsal raphe nucleus (DR) have been implicated in the facilitation of anxiety-related behavioral responses to anxiogenic drugs or aversive stimuli. In this study we attempted to determine a threshold to engage serotonergic neurons in the DR following exposure to aversive conditions in an anxiety-related behavioral test. We manipulated the intensity of anxiogenic stimuli in studies of male Wistar rats by leaving them undisturbed (CO), briefly handling them (HA), or exposing them to an open-field arena for 15-min under low-light (LL: 8-13 lx) or high-light (HL: 400-500 lx) conditions. Rats exposed to HL conditions responded with reduced locomotor activity, reduced time spent exploring the center of the arena, a lower frequency of rearing and grooming, and an increased frequency of facing the corner of the arena compared to LL rats. Rats exposed to HL conditions had small but significant increases in c-Fos expression within serotonergic neurons in subdivisions of the rostral DR. Exposure to HL conditions did not alter c-Fos responses in serotonergic neurons in any other DR subdivision. In contrast, rats exposed to the open-field arena had increased c-Fos expression in non-serotonergic cells throughout the DR compared to CO rats, and this effect was particularly apparent in the dorsolateral part of the DR. We conclude that exposure to HL conditions, compared to LL conditions, increased anxiety-related behavioral responses in an open-field arena but this stimulus was at or below the threshold required to increase c-Fos expression in serotonergic neurons.


Parental preference for one-stage versus two-stage surgical repair for children with congenital heart disease.
Little is known regarding parental preference for a one-stage complete repair versus a two-staged approach with initial palliation, followed by repair, of the congenital cardiac malformation. We interviewed 103 parents of healthy children referred to a clinic for pediatric cardiology. Participants were presented with a hypothetical scenario in which their children had a cardiac lesion requiring surgery. The surgery could be performed either by means of one-stage complete repair, or using a two-stage approach, with palliation first followed by complete repair a year later. The mortality rate for the one-stage repair was set at 5%. Participants were asked to choose between the one- and two-stage approaches, with differing mortality rates for the two-stage approach. The scenarios included options when the two-stage combined mortality rate was lower than the one-stage mortality, and the first stage mortality rate was at 1% and 3%, and when the two-stage combined mortality rate was the same as that for one-stage mortality, these being set at 1% and 3%. When the two-stage combined mortality rate was lower than that of the one-stage repair, participants were more likely to choose the two-stage approach if the first stage mortality rate was 1% as compared to 3% (57% and 44%, respectively, p = 0.04). When the two-stage combined mortality rate was the same as the one-stage approach, participants choosing the two-stage approach when the mortality rate was set at 1%, and when it was raised to 3%, were not significantly different (42% and 34%, respectively, p = 0.24). When the combined two-stage mortality was the same as that set for one-stage repair, participants with no insurance were less likely to choose the two-stage approach than those covered by insurance (p = 0.03). In the chosen scenarios, when the mortality for a two-stage combined approach is the same as that for one-staged repair, more parents choose the one-staged repair. If the two-stage combined mortality is lower than that for one-staged repair, parents are more likely to choose the two-stage repair if the mortality for the first stage is lower. When the mortality rates for the one-stage and two-stage approaches are the same, people without insurance are more likely to choose one-staged repair.


The effects of regurgitant orifice size, chamber compliance, and systemic vascular resistance on aortic regurgitant velocity slope and pressure half-time.
The determinants of the aortic regurgitant velocity profile have been investigated using computer and in vitro simulations in which regurgitant orifice area, ventricular and aortic compliance, and systemic vascular resistance could be independently varied. In the study, regurgitant fraction was altered, either by changing the size of the regurgitant orifice or by holding the regurgitant orifice constant and changing chamber compliance or systemic vascular resistance. Upon increasing regurgitant fraction by increasing the size of the regurgitant orifice, the slope got steeper and the pressure half-time shortened, the response anticipated in current clinical practice. However, when the regurgitant orifice was kept constant and regurgitation fraction was increased by increasing the systemic vascular resistance or by increasing the compliance of the left ventricle, slope became less steep and pressure half-time lengthened. Multivariate analysis was used to quantify the relationship of regurgitant fraction to slope and pressure half-time. When orifice area was allowed to vary, slope was related directly (multiple r = 0.78, p less than 0.001) and half-time was related inversely (multiple r = 0.66, p less than 0.001) to regurgitant fraction. With the orifice area fixed, however, directionally opposite responses were seen; slope varied inversely (multiple r = 0.87, p less than 0.001), whereas half-time varied directly (multiple r = 0.88, p less than 0.001) with regurgitant fraction. This study suggests that the utility of the slope and pressure half-time of the regurgitant velocity tracing in clinical practice relates to their ability to discriminate regurgitant orifices of differing sizes.(ABSTRACT TRUNCATED AT 250 WORDS)


Effectiveness of interactive videodisc instruction for the continuing education of paramedics.
To evaluate the effectiveness of interactive videodisc (IVD) instruction of paramedics through the use of computer analysis of trip sheets. Prospective, controlled, in an urban 9-1-1, paramedic, emergency medical services (EMS) system with total call volume of 62,000/year; 15,000 advanced life support (ALS). All 150 paramedics in the system received eight hours of IVD instruction covering five subject areas: 1) airway; 2) head/cervical trauma; 3) chest; 4) shock; and 5) cardiac arrest. Trip sheets from 9,943 runs in the pre-IVD period were subjected to computer analysis, and a compliance score was generated using previously developed algorithms that assigned a weight to each omission and commission. After a nine-month IVD training period, 4,303 cases were collected and analyzed in the post-IVD period. Statistical analyses were made using "Student's" t-test and Chi-square with alpha set at 0.05. Only those records of adult patients who fit one of the five protocols were eligible for computer analysis. Of the 9,943 cases in the pre-IVD group, 480 (4.8%) were excluded, all due to inadequate data recording by the paramedics. A statistically similar portion, 233 (5.4%) of the 4,303 post-IVD instruction cases were excluded (p = .15). Overall the mean compliance score of the pre-IVD group was 0.65 +/-0.19 (+/-SD). The post-IVD group score was 0.65 +/-0.19 (p = 0.99). Analysis of scores for each algorithm also showed no significant differences. This study had an observed power of .94 to detect a difference in compliance as small as 0.030. (ABSTRACT TRUNCATED AT 250 WORDS)


Enteropathogenic Escherichia coli translocate Tir and form an intimin-Tir intimate attachment to red blood cell membranes.
Type III secretion allows bacteria to inject effector proteins into host cells. In enteropathogenic Escherichia coli (EPEC) the type III secreted protein, Tir, is translocated to the host-cell plasma membrane where it functions as a receptor for the bacterial adhesin intimin, leading to intimate bacterial attachment and "attaching and effacing" (A/E) lesion formation. To study EPEC type III secretion the interaction of EPEC with monolayers of red blood cells (RBCs) has been exploited and in a recent study [Shaw, R. K., Daniell, S., Ebel, F., Frankel, G. & Knutton, S. (2001 ). Cell Microbiol 3, 213-222] it was shown that EPEC induced haemolysis of RBCs and translocation of EspD, a putative pore-forming type III secreted protein in the RBC membrane. Here it is demonstrated that EPEC are able to translocate and correctly insert Tir into the RBC membrane and produce an intimin-Tir intimate bacterial attachment, identical to that seen in A/E lesions. Following translocation Tir did not undergo any change in apparent molecular mass or become tyrosine-phosphorylated and there was no focusing of RBC cytoskeletal actin beneath intimately adherent bacteria, and no pedestal formation. This study, employing an RBC model of infection, has demonstrated that Tir translocation can be separated from host-cell-mediated Tir modifications; the data show that the EPEC type III protein translocation apparatus is sufficient to deliver and correctly insert Tir into host-cell membranes independent of eukaryotic cell functions.


GPVI and GPIbα mediate staphylococcal superantigen-like protein 5 (SSL5) induced platelet activation and direct toward glycans as potential inhibitors.
Staphylococcus aureus (S. aureus) is a common pathogen capable of causing life-threatening infections. Staphylococcal superantigen-like protein 5 (SSL5) has recently been shown to bind to platelet glycoproteins and induce platelet activation. This study investigates further the interaction between SSL5 and platelet glycoproteins. Moreover, using a glycan discovery approach, we aim to identify potential glycans to therapeutically target this interaction and prevent SSL5-induced effects. In addition to platelet activation experiments, flow cytometry, immunoprecipitation, surface plasmon resonance and a glycan binding array, were used to identify specific SSL5 binding regions and mediators. We independently confirm SSL5 to interact with platelets via GPIbα and identify the sulphated-tyrosine residues as an important region for SSL5 binding. We also identify the novel direct interaction between SSL5 and the platelet collagen receptor GPVI. Together, these receptors offer one mechanistic explanation for the unique functional influences SSL5 exerts on platelets. A role for specific families of platelet glycans in mediating SSL5-platelet interactions was also discovered and used to identify and demonstrate effectiveness of potential glycan based inhibitors in vitro. These findings further elucidate the functional interactions between SSL5 and platelets, including the novel finding of a role for the GPVI receptor. We demonstrate efficacy of possible glycan-based approaches to inhibit the SSL5-induced platelet activation. Our data warrant further work to prove SSL5-platelet effects in vivo.


Preclinical studies of the pan-Bcl inhibitor obatoclax (GX015-070) in multiple myeloma.
Bcl family members Bcl-2, Bcl-x(L), and Mcl-1, are frequently expressed and implicated in the survival of myeloma cells. Obatoclax (GX015-070) is a novel, small-molecule antagonist of the BH3-binding groove of the Bcl family of proteins. We show that GX015-070 inhibits the binding of Bak to Mcl-1, up-regulates Bim, induces cytochrome c release, and activates capase-3 in human myeloma cell lines (HMCLs), confirming the predicted mechanism of action. Consequently, GX015-070 potently inhibited the viability of 15 of 16 HMCLs (mean IC(50) of 246 nM), including those resistant to melphalan and dexamethasone. In combination studies, GX015-070 enhanced the antimyeloma activity induced by melphalan, dexamethasone, or bortezomib. Sensitivity to GX015-070 correlated with the absence or near absence of Bcl-x(L). Coculture with interleukin-6 or adherence to bone marrow stroma conferred modest resistance; however, it did not overcome GX015-070-induced cytotoxicity. Of importance, GX015-070 as a single agent induced potent cytotoxic responses against patient-derived tumor cells. GX015-070 inhibited normal bone marrow-derived colony formation; however, cytotoxicity to human blood lymphocytes was not observed. Taken together, these studies describe a novel BH3 mimic with selectivity for Mcl-1, and support the therapeutic application of GX015-070 for diverse neoplasias including multiple myeloma.


Contact activation of blood coagulation: trigger properties and hysteresis. Kinetic recognition of foreign surfaces upon contact activation of blood coagulation: a hypothesis.
A mathematical model of contact activation of blood coagulation was developed and analysed. The model variables are concentrations of factor XIIa, kallikrein and activated high-molecular-weight kininogen. Concentrations of active factors were shown to depend on the activating signal value in a hysteretic manner. Within a range of relatively small signals, two (activated and non-activated) stable states coexist (bistability). Signals of the natural environment (surfaces of endothelial and blood cells) seem to be in the range of bistability; therefore, contact activation that persists for a short time can induce a transition of the system to the activated state, and, correspondingly, the formation of a clot. The system cannot return to the initial state, which is characterized by low activation levels, until the activating signals decrease significantly below those present in the circulation.


Holothuria leucospilota Extract Induces Apoptosis in Leishmania major Promastigotes.
The present study aimed to survey antileishmanial activity of methanolic Holothuria leucospilota extract against Leishmania major promastigotes in vitro. Promastigotes were cultured in RPMI 1640 and after reaching the stationary phase, the study was conducted with different concentrations of the extract. Afterwards, MTT colorimetric assay for the obtaining of 50% inhibitory concentration (IC50) was utilized. Furthermore, in order to determine the possible induction of apoptosis in L. major promastigotes, flow cytometry and DNA fragmentation methods were employed using annexin-V FLUOS staining kit and DNA ladder kit, respectively. The IC50 value of H. leucospilota extract at three time points of 24, 48, and 72 h was estimated 2000, 300 and 85 μg/ml, respectively. In addition, the extract revealed a dose and time-dependent antileishmanial activity. Furthermore, various characteristics of apoptosis appeared after L. major promastigotes treatment, which included cell shrinkage, formation of apoptotic bodies, blebbing of the cell membrane, and externalization of phosphatidylserine, although no laddering pattern was observed. The methanolic extract of H. leucospilota possesses lethal effect on L. major promastigotes and induces the apoptosis in parasites. Further studies are required to address the apoptosis mechanism in vivo.


Rates of colon and rectal cancers are increasing in young adults.
Incidence rates for colorectal cancer are decreasing in the United States, possibly due to preventative cancer screening. Because these programs target older patients, their beneficial effects may not apply to young patients. The purpose of this study was to compare incidence rates and tumor characteristics of colon and rectal cancers for young versus older patients using a population-based cancer registry. Colon and rectal cancer patients reported in the Surveillance, Epidemiology, and End Results registry (1973-1999) were separately analyzed. Incidence rates over time, stage, and grade were compared for two age groups: young patients (20-40 years, n = 5383) and older patients (60+ years, n = 256,401). For older patients, colon cancer incidence remained stable while rectal cancer incidence decreased 11 per cent to 72.1/100,000 persons (P < 0.05). For the young, colon cancer incidence increased 17 per cent to 2.1 (P < 0.05), and rectal incidence rose 75 per cent to 1.4 (P < 0.05). Young patients had less localized tumors than older patients: colon (25.8% vs. 35.3%, P < 0.001); rectal (38.4% vs. 41.7%, P = 0.005). Young patients also had more poorly differentiated tumors: colon (22.2% vs. 14.7%, P < 0.001); rectal (16.4% vs. 12.3%, P < 0.001). Incidence rates for colon and rectal cancers in young patients are rising, and they have more advanced disease. Although the overall prevalence is low in this population, the increasing incidence suggests health-care providers should have heightened awareness when caring for this population.


The AKT/NF-κB inhibitor xanthohumol is a potent anti-lymphocytic leukemia drug overcoming chemoresistance and cell infiltration.
Although the vast majority of patients with acute lymphocytic leukemia (ALL) attain remission with modern therapies, relapsed leukemia will continue to be a common malignancy both in childhood and in adults, until new treatments are available. Therapeutic options for advanced B-cell acute lymphocytic leukemia are still limited and acquired drug resistance and extramedullary tissue infiltration are two major obstacles during treatment. The prenylflavonoid xanthohumol (XN) has shown in vitro and in vivo therapeutic potential against a range of tumors. In the present study we investigated the effects of XN on B-ALL cells in vitro and in an ALL-like xenograft mouse model. Treatment of ALL cell lines with XN resulted in growth arrest and apoptosis induction. XN retained its cytotoxicity when adriamycin resistant cells were examined while ALL cell clones adapted to long-term exposure to XN resulted highly responsive to cytotoxic drugs. Administration of 50μg XN/mouse (5 days/week) significantly increased animal life span by delaying the insurgence of neurological disorders due to leukemic cells dissemination. In agreement with a less invasive phenotype, cell migration and invasion were impaired by XN and basal levels of FAK, AKT and NF-κB signaling pathways were down-regulated in ALL cells upon XN exposure. Our data indicate that XN has significant antileukemic activity both in vitro and in vivo, which associates with impaired cell migration and invasion. Interestingly, this activity overcomes mechanisms leading to drug-resistance. XN represents a promising agent perspective for ALL therapy and recurrence prevention and would deserve clinical testing in the near future.


Dental trauma and prevention guidelines: a narrative review.
Dental injuries affect different patients every day in conditions that can range from domestic, work, or road accidents. As for traumas in the age of development, the field is restricted to domestic, sports, and school environments. The purpose of this study was to clarify the current protocols in the literature to limit and manage this type of pathology. This narrative review considers the literature of the last 20 years on this topic in different ways. The literature is in agreement with dividing the treatments into primary and secondary and also according to what is the place where the trauma occurred to evaluate the type of intervention. However, all protocols are aimed at implementing efficient preventive strategies rather than having to solve a problem subsequently, certainly, new protocols and protection systems can limit this problem which leads not only to more or less complex problems related to oral health and aesthetics but also possible subsequent psychological problems.


Validity of a laser fluorescence system (DIAGNOdent) for detection of occlusal caries in third molars: an in vitro study.
The aim of this study was to compare the validity of the measurements of the laser fluorescence device, KaVo DIAGNOdent, with the result of polarized light microscopy in the detection of occlusal fissure caries in extracted third molars. Ten impacted, surgically removed, and 25 extracted third molars with macroscopically intact occlusal surfaces were selected. The DIAGNOdent measurements of the occlusal test site were recorded by two observers at intervals of 2 days. The teeth were then sectioned at the specified test sites for histological examinations. Prepared specimens were evaluated under the polarized light microscopy and all images were scored with the caries classification of D1 (sound and fissure lesion in the half of the outer enamel), D2 (enamel decay) and D3 (dentin decay) level (gold standard). The kappa value for the inter-observer repeatability was calculated and the value 0.83 for the first measurements and 0.67 for the second measurement were obtained,respectively. Inter-observer correct classification percentages were 88.5 and 77.1 for the first and second measurements, respectively. The kappa value for intra-observer repeatability was 0.79 for the first observer and 0.75 for the second observer. Intra-observer correct classification percentage values were 85.7 and 82.8 for the first and second observer, respectively. Value of specificity for the detection of enamel caries at D1 level was 0.74 and sensitivity values at D2 and D3 levels were 0.66 and 1.00, respectively. The present study indicates that the DIAGNOdent provides not only almost perfect agreement but also sufficient repeatability at D1, D2, D3 levels and better specificity at D1 level as well as lower sensitivity at D2 level and excellent sensitivity at D3 level.


Uncoupling of PARP1 trapping and inhibition using selective PARP1 degradation.
PARP1 inhibitors (PARPi) are known to kill tumor cells via two mechanisms (PARP1 catalytic inhibition and PARP1 trapping). The relative contribution of these two pathways in mediating the cytotoxicity of PARPi, however, is not well understood. Here we designed a series of small molecule PARP degraders. Treatment with one such compound iRucaparib-AP6 results in highly efficient and specific PARP1 degradation. iRucaparib-AP6 blocks the enzymatic activity of PARP1 in vitro, and PARP1-mediated poly-ADP-ribosylation signaling in intact cells. This strategy mimics PARP1 genetic depletion, which enables the pharmacological decoupling of PARP1 inhibition from PARP1 trapping. Finally, by depleting PARP1, iRucaparib-AP6 protects muscle cells and primary cardiomyocytes from DNA-damage-induced energy crisis and cell death. In summary, these compounds represent 'non-trapping' PARP1 degraders that block both the catalytic activity and scaffolding effects of PARP1, providing an ideal approach for the amelioration of the various pathological conditions caused by PARP1 hyperactivation.


Theoretical perspectives on the relation between catastrophizing and pain.
The tendency to "catastrophize" during painful stimulation contributes to more intense pain experience and increased emotional distress. Catastrophizing has been broadly conceived as an exaggerated negative "mental set" brought to bear during painful experiences. Although findings have been consistent in showing a relation between catastrophizing and pain, research in this area has proceeded in the relative absence of a guiding theoretical framework. This article reviews the literature on the relation between catastrophizing and pain and examines the relative strengths and limitations of different theoretical models that could be advanced to account for the pattern of available findings. The article evaluates the explanatory power of a schema activation model, an appraisal model, an attention model, and a communal coping model of pain perception. It is suggested that catastrophizing might best be viewed from the perspective of hierarchical levels of analysis, where social factors and social goals may play a role in the development and maintenance of catastrophizing, whereas appraisal-related processes may point to the mechanisms that link catastrophizing to pain experience. Directions for future research are suggested.


Insights from the reanalysis of high-throughput chemical genomics data for Escherichia coli K-12.
Despite the demonstrated success of genome-wide genetic screens and chemical genomics studies at predicting functions for genes of unknown function or predicting new functions for well-characterized genes, their potential to provide insights into gene function has not been fully explored. We systematically reanalyzed a published high-throughput phenotypic dataset for the model Gram-negative bacterium Escherichia coli K-12. The availability of high-quality annotation sets allowed us to compare the power of different metrics for measuring phenotypic profile similarity to correctly infer gene function. We conclude that there is no single best method; the three metrics tested gave comparable results for most gene pairs. We also assessed how converting quantitative phenotypes to discrete, qualitative phenotypes affected the association between phenotype and function. Our results indicate that this approach may allow phenotypic data from different studies to be combined to produce a larger dataset that may reveal functional connections between genes not detected in individual studies.


Effect on blood pressure of potassium, calcium, and magnesium in women with low habitual intake.
In populations, dietary intakes of potassium, calcium, and magnesium each have been inversely associated with blood pressure. However, most clinical trials in normotensive populations have not found that dietary supplements of these minerals lowered blood pressure. We tested the hypothesis that normotensive persons who have low habitual intake of these minerals would be particularly responsive to supplementation. Three hundred normotensive women in the Nurses Health Study II (mean age, 39 years), whose reported intakes of potassium, calcium, and magnesium were between the 10th and 15th percentiles, received for 16 weeks' duration daily supplements of either potassium 40 mmol, calcium 30 mmol (1200 mg), magnesium 14 mmol (336 mg), all three minerals together or placebos. At baseline, mean (+/-SD) 24-hour ambulatory blood pressures were 116+/-8 and 73+/-6 mm Hg systolic and diastolic, respectively, and mean dietary intakes of potassium, calcium, and magnesium were 62+/-20 mmol/d, 638+/-265 mg/d, and 239+/-79 mg/d, respectively. The mean differences (with 95% confidence intervals) of the changes in systolic and diastolic blood pressures between the treatment and placebo groups were significant for potassium, -2.0 (-3.7 to -0.3) and -1.7 (-3.0 to -0.4), but not for calcium, -0.6 (-2.2 to 1.0) and -0.7 (-2.0 to 0.6), or for magnesium, -0.9 (-2.6 to 0.8) and -0.7 (-2.2 to 0.8). The administration of calcium and magnesium with potassium did not enhance the effect of potassium alone, and the changes in blood pressure were not significant -1.3 (-3.0 to 0.4) and -0.9 (-2.2 to 0.4). In conclusion, potassium, but not calcium or magnesium supplements, has a modest blood pressure-lowering effect in normotensive persons with low dietary intake. This study strengthens evidence for the importance of potassium for blood pressure regulation in the general population.


Histone deacetylase inhibition modulates histone acetylation at gene promoter regions and affects genome-wide gene transcription in Schistosoma mansoni.
Schistosomiasis is a parasitic disease infecting hundreds of millions of people worldwide. Treatment depends on a single drug, praziquantel, which kills the Schistosoma spp. parasite only at the adult stage. HDAC inhibitors (HDACi) such as Trichostatin A (TSA) induce parasite mortality in vitro (schistosomula and adult worms), however the downstream effects of histone hyperacetylation on the parasite are not known. TSA treatment of adult worms in vitro increased histone acetylation at H3K9ac and H3K14ac, which are transcription activation marks, not affecting the unrelated transcription repression mark H3K27me3. We investigated the effect of TSA HDACi on schistosomula gene expression at three different time points, finding a marked genome-wide change in the transcriptome profile. Gene transcription activity was correlated with changes on the chromatin acetylation mark at gene promoter regions. Moreover, combining expression data with ChIP-Seq public data for schistosomula, we found that differentially expressed genes having the H3K4me3 mark at their promoter region in general showed transcription activation upon HDACi treatment, compared with those without the mark, which showed transcription down-regulation. Affected genes are enriched for DNA replication processes, most of them being up-regulated. Twenty out of 22 genes encoding proteins involved in reducing reactive oxygen species accumulation were down-regulated. Dozens of genes encoding proteins with histone reader motifs were changed, including SmEED from the PRC2 complex. We targeted SmEZH2 methyltransferase PRC2 component with a new EZH2 inhibitor (GSK343) and showed a synergistic effect with TSA, significantly increasing schistosomula mortality. Genome-wide gene expression analyses have identified important pathways and cellular functions that were affected and may explain the schistosomicidal effect of TSA HDACi. The change in expression of dozens of histone reader genes involved in regulation of the epigenetic program in S. mansoni can be used as a starting point to look for possible novel schistosomicidal targets.


From abortion to contraception in Europe.
This review summarizes abortion statistics, mainly in Europe and primarily in Eastern Europe. The reasons for unwanted pregnancies and consequent abortions are analyzed. Recommendations are proposed to achieve a shift from abortion to contraception in fertility regulation. The relevant literature and the results of the data collected by the author in 16 Eastern European countries by means of a questionnaire are reviewed. The need and the possibilities for better future perspectives are outlined in the conclusions of The Szeged Declaration. The major problems identified were the low level or lack of training of professionals, deficiencies in the knowledge of current family planning methods among professionals, consumers, policy makers and media representatives, the lack of supplies of modern contraceptives to meet the needs of the population, the lack of sex education in schools and the high level of induced abortions. In the majority of the countries in the Eastern European region, there is a need for the establishment of a system for the delivery of high-quality family planning services. This imposes a requirement for the establishment of a system for the continuous supply of contraceptives, through national family planning programs, by governmental and/or nongovernmental organizations. To increase contraceptive use, countries in this region should consider the establishment of extensive educational programs on current family planning methods and other aspects of reproductive health for their health professionals, consumers and representatives of the media. Sex education in schools should be promoted. Governments should be encouraged to invest more in setting up and running family planning services and programs.


Determination of gas phase protein ion densities via ion mobility analysis with charge reduction.
We use a charge reduction electrospray (ESI) source and subsequent ion mobility analysis with a differential mobility analyzer (DMA, with detection via both a Faraday cage electrometer and a condensation particle counter) to infer the densities of single and multiprotein ions of cytochrome C, lysozyme, myoglobin, ovalbumin, and bovine serum albumin produced from non-denaturing (20 mM aqueous ammonium acetate) and denaturing (1 : 49.5 : 49.5, formic acid : methanol : water) ESI. Charge reduction is achieved through use of a Po-210 radioactive source, which generates roughly equal concentrations of positive and negative ions. Ions produced by the source collide with and reduce the charge on ESI generated drops, preventing Coulombic fissions, and unlike typical protein ESI, leading to gas-phase protein ions with +1 to +3 excess charges. Therefore, charge reduction serves to effectively mitigate any role that Coulombic stretching may play on the structure of the gas phase ions. Density inference is made via determination of the mobility diameter, and correspondingly the spherical equivalent protein volume. Through this approach it is found that for both non-denaturing and denaturing ESI-generated ions, gas-phase protein ions are relatively compact, with average densities of 0.97 g cm(-3) and 0.86 g cm(-3), respectively. Ions from non-denaturing ESI are found to be slightly more compact than predicted from the protein crystal structures, suggesting that low charge state protein ions in the gas phase are slightly denser than their solution conformations. While a slight difference is detected between the ions produced with non-denaturing and denaturing ESI, the denatured ions are found to be much more dense than those examined previously by drift tube mobility analysis, in which charge reduction was not employed. This indicates that Coulombic stretching is typically what leads to non-compact ions in the gas-phase, and suggests that for gas phase measurements to be correlated to biomolecular structures in solution, low charge state ions should be analyzed. Further, to determine if different solution conditions give rise to ions of different structure, ions of similar charge state should be compared. Non-denatured protein ion densities are found to be in excellent agreement with non-denatured protein ion densities inferred from prior DMA and drift tube measurements made without charge reduction (all ions with densities in the 0.85-1.10 g cm(-3) range), showing that these ions are not strongly influenced by Coulombic stretching nor by analysis method.


Na+- and K+-dependent oligomeric interconversion among alphabeta-protomers, diprotomers and higher oligomers in solubilized Na+/K+-ATPase.
Protein fractions of a higher-oligomer (H), (alphabeta)(2)-diprotomer (D) and alphabeta-protomer (P) were separated from dog kidney Na(+)/K(+)-ATPase solubilized in the presence of NaCl and KCl. Na(+)/K(+)-dependent interconversion of the oligomers was analysed using HPLC at 0 degrees C. With increasing KCl concentrations, the content or amount of D increased from 27.6 to 54.3% of total protein, i.e. DeltaC(max) = 26.7%. DeltaC(max) for the sum of D and H was equivalent to the absolute value of DeltaC(max) for P, regardless of the anion present, indicating that K(+) induced the conversion of P into D and/or H, and Na(+) had the opposite effect. When enzymes that had been denatured to varying degrees by aging were solubilized, DeltaC(max) increased linearly with the remaining ATPase activity. The magnitude of the interconversion could be explained based on an equilibrium of D <==> 2P, assuming 50-fold difference in the K(d) between KCl and NaCl, and coexistence of unconvertible oligomers, which comprised as much as 39% of the eluted protein. Oligomeric interconversion, determined as a function of the KCl or NaCl concentration, showed K(0.5)s of 64.8 microM and 6.50 mM for KCl and NaCl, respectively, implying that oligomeric interconversion was coupled with Na(+)/K(+)-binding to their active transport sites.


Systemic lupus erythematosus. Some contemporary laboratory aspects.
A summary of some of the important laboratory features of systemic lupus erythematosus (SLE) is presented. Emphasis is given to the sensitivity and specificity of the antinuclear antibody test, the relevance of particular antinuclear antibody patterns, and the status of testing for antibodies to DNA. Serum complement abnormalities in the patient with SLE are briefly reviewed, and the utility of the total hemolytic complement assay is stressed. The increasing evidence for disordered immune regulation in SLE is considered. Particular attention is given to the role of antilymphocyte antibodies and in vitro assays that permit an assessment of functional lymphocyte subsets. The heterogeneity of the SLE patient population and the influence of treatment and/or disease activity on laboratory results is repeatedly emphasized.


Arab and Jewish elementary school students' perceptions of fear and school violence: understanding the influence of school context.
This inquiry explores variables that predict elementary school students' fear of attending school due to school violence and their overall judgments of school violence as a problem. Using a nationally representative sample (Israel) of 5,472 elementary-school-aged children, this study tested the hypotheses that: (a) young students' personal fear of attending school due to violence, and (b) students' assessment of a school violence problem, are best understood as separate conceptual constructs. Structural equation modelling was used to test the proposed theoretical model for the sample as a whole and separately for across gender and for Arab and Jewish students. Student fear of attending school due to violence was related directly to experiences of personal victimization on school grounds by students and teachers. Children's judgments of their schools' overall violence problem were influenced directly by the school climate, risky peer-group behaviours, and personal victimization. The findings provide evidence that the proposed theoretical model applies across gender groups and for both Arab and Jewish students. Implications for policy, theory, and future research are highlighted.


Efficacy of pegylated interferon plus ribavirin combination therapy for hepatitis C patients with normal ALT levels: a matched case-control study.
The antiviral effect of pegylated interferon (Peg-IFN) plus ribavirin combination therapy in chronic hepatitis C (CHC) patients with normal alanine aminotransferase (ALT) levels (N-ALT) has been reported to be equivalent to that for patients with elevated ALT levels (E-ALT). However, the actual antiviral effect in N-ALT patients remains obscure because efficacy can be overestimated in patients with an advantageous background. In this study, 386 patients were extracted, for a matched case-control study, from 1320 CHC patients treated with Peg-IFN alpha-2b plus ribavirin combination therapy; 193 N-ALT patients [116 with hepatitis C virus genotype 1 (HCV-1), 77 with HCV genotype 2 (HCV-2)] were matched with 193 E-ALT patients by a propensity score method using the variables of age, sex, IFN treatment history, body mass index, and platelet counts. On multivariate analysis for sustained virological response (SVR) in N-ALT patients, younger age, low HCV RNA level at baseline, and HCV-2 were significant factors. The matched case-control study showed that the SVR rates of N-ALT patients were equivalent to those of E-ALT patients; at 49 and 40% in the HCV-1 group (P = 0.146), and 78 and 81% in the HCV-2 group (P = 0.691). However, in N-ALT patients with non-SVR, approximately 40% showed ALT elevation at 24 weeks post-treatment. Our findings indicate that the antiviral effect of Peg-IFN plus ribavirin therapy in N-ALT patients is comparable to that for E-ALT patients irrespective of their advantageous background; however, the application of this therapy for N-ALT patients, especially for those with HCV-1, should be considered carefully.


Key importance of the Helicobacter pylori adherence factor blood group antigen binding adhesin during chronic gastric inflammation.
Helicobacter pylori has been assigned as a class I carcinogen because of its relation to gastric adenocarcinoma. Chronic H. pylori infection may lead to severe gastritis, glandular atrophy (AT), and intestinal metaplasia (IM). Strains secreting the vacuolating toxin VacA and producing the cytotoxin-associated antigen CagA (type 1 strains), as well as the blood group antigen binding adhesin (BabA) targeting Lewis(b) antigens, have been associated previously with distal gastric adenocarcinoma (M. Gerhard et al., Proc. Natl. Acad. Sci. USA, 96: 12778-12783, 1999) and may therefore also be related to lesions preceding gastric cancer. Antral and corpus biopsies were collected from 451 patients; 151 were H. pylori positive, as determined by PCR. Gastric biopsies were histologically evaluated for activity of gastritis (G0-G3, granulocyte infiltration), chronicity of gastritis (L1-L3, lymphocyte infiltration), and the presence of IM and/or AT according to the Sydney classification. Simultaneously, the presence of bacterial genes encoding virulence and adherence factors (racAs1/s2, cagA, and babA2) was determined by PCR. The presence of cagA+ and vacAs1 (alone or combined) both correlated with activity and chronicity of gastritis (P < 0.05); however, the overall prevalence of these genes was 60 or 72%, respectively, and was thus relatively frequent. The babA2 gene, encoding the adhesin BabA, was detected in 38% of infected patients and was correlated with the activity of gastritis in antrum and corpus (P < 0.005). cagA+/vacAs1+ strains (suggesting the presence of type 1 strains) that were also babA2 positive were detected more frequently in patients with severe histological alterations (such as G3, IM, or AT) compared with subjects without these changes (P < 0.01). cagA+/vacAs1+ strains that were babA2 negative, however, lacked a significant correlation with severe histological changes, activity, or chronicity of gastritis in antrum and corpus. Adherence of H. pylori via BabA appears to be of importance for efficient delivery of VacA and CagA and may play a special role in the pathogenesis of severe histological changes.


[Structure of the longitudinal suture of the bony base of the hard palate during the period of permanent bite formation].
The investigations performed support the literature data that the longitudinal connective tissue suture in the osseous base of the hard palate during formation of the permanent bite possesses all the necessary tissue components in order to realize certain osteogenic functions. Its blood supply is provided at the expense of the microcirculatory bed including arterioles, precapillaries, capillaries and postcapillaries. The main morphological signs, demonstrating active osteogenic processes in the connective tissue suture of the hard palate, are preserved during the whole period when the permanent bite is forming; they remain more or less constant, while the form of the connective tissue suture of the hard palate undergoes certain rearrangements. It is necessary to note that among tissue components of the longitudinal connective tissue suture of the hard palate, besides osteoblasts and osteoclasts, Sharpey's fibres are important constructive elements. In addition to their mechanical function, they are capable to participate in transport of fluid and dissolved substances from the interstitium of the connective tissue suture into the osseous tissue along peculiar tubules in which the Sharpey's fibres are situated.


Absence of filamin A prevents cells from responding to stiffness gradients on gels coated with collagen but not fibronectin.
Cell types from many tissues respond to changes in substrate stiffness by actively remodeling their cytoskeletons to alter spread area or adhesion strength, and in some cases changing their own stiffness to match that of their substrate. These cell responses to substrate stiffness are linked to substrate-induced changes in the state, localization, and amount of numerous proteins, but detailed evidence for the requirement of specific proteins in these distinct forms of mechanical response are scarce. Here we use microfluidics techniques to produce gels with a gradient of stiffness to show the essential function of filamin A in cell responses to mechanical stimuli and dissociate cell spreading and stiffening by contrasting responses of a pair of human melanoma-derived cell lines that differ in expression of this actin cross-linking protein. M2 melanoma cells null for filamin A do not alter their adherent area in response to increased substrate stiffness when they link to the substrate only through collagen receptors, but change adherent area normally when bound through fibronectin receptors. In contrast, filamin A-replete A7 cells change adherent area on both substrates and respond more strongly to collagen I-coated gels than to fibronectin-coated gels. Strikingly, A7 cells alter their stiffness, as measured by atomic force microscopy, to match the elastic modulus of the substrate immediately adjacent to them on the gradient. M2 cells, in contrast, maintain a constant stiffness on all substrates that is as low as that of A7 cells on the softest gels examined (1000 Pa). Comparison of cell spreading and cell stiffening on the same gradient substrates shows that cell spreading is uncoupled from stiffening. At saturating collagen and fibronectin concentrations, adhesion of M2 cells is reduced compared to that of A7 cells to an extent approximately equal to the difference in adherent area. Filamin A appears to be essential for cell stiffening on collagen, but not for cell spreading on fibronectin. These results have implications for different models of cell protrusion and adhesion and identify a key role for filamin A in altering cellular stiffness that cannot be compensated for by other actin cross-linkers in vivo.


Quantifying protein interactions with isomeric carbohydrate ligands using a catch and release electrospray ionization-mass spectrometry assay.
The application of a catch-and-release electrospray ionization mass spectrometry (CaR-ESI-MS) assay to quantify interactions between proteins and isomeric carbohydrate ligands is described. Absolute affinities for each ligand are determined from the abundance ratio of ligand-bound to free protein measured directly by ESI-MS and the relative abundances of the individual isomeric ligands, which are established by releasing the ligands, in their deprotonated form, from the protein using collision-induced dissociation (CID) and subjecting them to ion mobility separation (IMS) or another stage of CID to fragment the ions. Using Gaussian functions to represent the contributions of individual ligands to the arrival time distributions (ATDs) measured by IMS, the relative abundance of each ligand bound to the protein can be established. A modification of this method, suitable for cases where nonspecific ligand-protein binding occurs during the ESI process, is also described. In cases where the ATDs are not sufficiently different to distinguish the isomeric ligands, CID can establish the relative abundance of each ligand bound to the protein from the relative abundance of the resulting fragment ions. The implementation and reliability of the CaR-ESI-MS assay for the analysis of isomeric carbohydrate ligands is demonstrated using three carbohydrate-binding proteins, a single chain antibody, an antigen binding fragment, and a fragment of a bacterial toxin, and their interactions with isomeric carbohydrate ligands with affinities ranging from 10(3) to 10(5) M(-1).


Fast detection and structural identification of carbocations on zeolites by dynamic nuclear polarization enhanced solid-state NMR.
Acidic zeolites are porous aluminosilicates used in a wide range of industrial processes such as adsorption and catalysis. The formation of carbocation intermediates plays a key role in reactivity, selectivity and deactivation in heterogeneous catalytic processes. However, the observation and determination of carbocations remain a significant challenge in heterogeneous catalysis due to the lack of selective techniques of sufficient sensitivity to detect their low concentrations. Here, we combine 13C isotopic enrichment and efficient dynamic nuclear polarization magic angle spinning nuclear magnetic resonance spectroscopy to detect carbocations in zeolites. We use two dimensional 13C-13C through-bond correlations to establish their structures and 29Si-13C through-space experiments to quantitatively probe the interaction between multiple surface sites of the zeolites and the confined hydrocarbon pool species. We show that a range of various membered ring carbocations are intermediates in the methanol to hydrocarbons reaction catalysed by different microstructural β-zeolites and highlight that different reaction routes for the formation of both targeted hydrocarbon products and coke exist. These species have strong van der Waals interaction with the zeolite framework demonstrating that their accumulation in the channels of the zeolites leads to deactivation. These results enable understanding of deactivation pathways and open up opportunities for the design of catalysts with improved performances.


5-year survival and rehospitalization due to stroke recurrence among patients with hemorrhagic or ischemic strokes in Singapore.
Stroke is the 4th leading cause of death and 1st leading cause of disability in Singapore. However the information on long-term post stroke outcomes for Singaporean patients was limited. This study aimed to investigate the post stroke outcomes of 5-year survival and rehospitalization due to stroke recurrence for hemorrhagic and ischemic stroke patients in Singapore. The outcomes were stratified by age, ethnic group, gender and stroke types. The causes of death and stroke recurrence were also explored in the study. A multi-site retrospective cohort study. Patients admitted for stroke at any of the three hospitals in the National Healthcare Group of Singapore were included in the study. All study patients were followed up to 5 years. Kaplan-Meier was applied to study the time to first event, death or rehospitalization due to stroke recurrence. Cox proportional hazard model was applied to study the time to death with adjustment for stroke type, age, sex, ethnic group, and admission year. Cumulative incidence model with competing risk was applied for comparing the risks of rehospitalization due to stroke recurrence with death as the competing risk. Totally 12,559 stroke patients were included in the study. Among them, 59.3% survived for 5 years; 18.4% were rehospitalized due to stroke recurrence in 5 years. The risk of stroke recurrence and mortality increased with age in all stroke types. Gender, ethnic group and admitting year were not significantly associated with the risk of mortality or stroke recurrence in hemorrhagic stroke. Male or Malay patient had higher risk of stroke recurrence and mortality in ischemic stroke. Hemorrhagic stroke had higher early mortality while ischemic stroke had higher recurrence and late mortality. The top cause of death among died stroke patients was cerebrovascular diseases, followed by pneumonia and ischemic heart diseases. The recurrent stroke was most likely to be the same type as the initial stroke among rehospitalized stroke patients. Five year post-stroke survival and rehospitalization due to stroke recurrence as well as their associations with patient demographics were studied for different stroke types in Singapore. Specific preventive strategies are needed to target the high risk groups to improve their long-term outcomes after acute stroke.


Whole-Genome Screening of Newborns? The Constitutional Boundaries of State Newborn Screening Programs.
State newborn screening (NBS) programs routinely screen nearly all of the 4 million newborns in the United States each year for ∼30 primary conditions and a number of secondary conditions. NBS could be on the cusp of an unprecedented expansion as a result of advances in whole-genome sequencing (WGS). As WGS becomes cheaper and easier and as our knowledge and understanding of human genetics expand, the question of whether WGS has a role to play in state NBS programs becomes increasingly relevant and complex. As geneticists and state public health officials begin to contemplate the technical and procedural details of whether WGS could benefit existing NBS programs, this is an opportune time to revisit the legal framework of state NBS programs. In this article, we examine the constitutional underpinnings of state-mandated NBS and explore the range of current state statutes and regulations that govern the programs. We consider the legal refinements that will be needed to keep state NBS programs within constitutional bounds, focusing on 2 areas of concern: consent procedures and the criteria used to select new conditions for NBS panels. We conclude by providing options for states to consider when contemplating the use of WGS for NBS.


Periodontal parameters of two types of 3 x 3 orthodontic retainer: a longitudinal study.
bonded fixed retainers are often used to stabilize the results obtained with the orthodontic treatment. It is important that they do not prejudice dental health, as they will be used for a long period. The purpose of the present study was to compare periodontal indexes between two types of bonded fixed retainers, conventional 3 x 3 plain retainer (0.8-mm orthodontic wire, bonded to the canines only) and a manufactured braided retainer (0.2 x 0.7-mm stainless steel wire, bonded to all anterior teeth) after use. a test group of 15 volunteers (aged from 18 to 25 years) used both the conventional retainer and braided retainer for six months. A randomized longitudinal study design, with a two week washout interval, was applied. The dental plaque index, gingival index and dental calculus index were evaluated. Furthermore, the calculus accumulated along the retainer wire was measured and all patients answered a questionnaire about the use, acceptance and comfort of both types of retainers. the scores for plaque and gingival indexes were higher for the braided retainer (p< 0.05) on the lingual and proximal surfaces. The same occurred with the calculus index on the lingual surfaces (p< 0.05). The calculus index along wire was higher for the braided retainer (p< 0.05). All patients preferred the conventional retainer, and said that it was also more comfortable to use. it was concluded that the conventional retainer showed better periodontal indexes than the braided type.


Human HLA-DR transgenes protect mice from fatal virus-induced encephalomyelitis and chronic demyelination.
We evaluated the participatory role of human HLA-DR molecules in control of virus from the central nervous system and in the development of subsequent spinal cord demyelination. The experiments utilized intracranial infection with Theiler's murine encephalomyelitis virus (TMEV), a picornavirus that, in some strains of mice, results in primary demyelination. We studied DR2 and DR3 transgenic mice that were bred onto a combined class I-deficient mouse (beta-2 microglobulin deficient; beta2m(0)) and class II-deficient mouse (Abeta(0)) of the H-2(b) background. Abeta(0).beta2m(0) mice infected with TMEV died within 18 days of infection. These mice showed severe encephalomyelitis due to rapid replication of virus genome. In contrast, transgenic mice with insertion of a single human class II major histocompatibility complex (MHC) gene (DR2 or DR3) survived the acute infection. DR2 and DR3 mice controlled virus infection by 45 days and did not develop spinal cord demyelination. Levels of virus RNA were reduced in HLA-DR transgenic mice compared to Abeta(0).beta2m(0) mice. Virus-neutralizing antibody responses did not explain why DR mice survived the infection and controlled virus replication. However, DR mice showed an increase in gamma interferon and interleukin-2 transcripts in the brain, which were associated with protection. The findings support the hypothesis that the expression of a single human class II MHC molecule can, by itself, influence the control of an intracerebral pathogen in a host without a competent class I MHC immune response. The mechanism of protection appears to be the result of cytokines released by CD4(+) T cells.


Multidisciplinary Team Treatment Approaches to Chronic Daily Headaches.
In this review, we focus on nonmedication treatment approaches to chronic daily headaches and chronic migraine. We review the current scientific data on studies using multimodal treatments, especially physical therapy and occupational therapy, and provide recommendations on the formation of interdisciplinary headache teams. Chronic daily headache, which includes chronic migraine, is a particularly challenging clinical entity which often involves multiple headache types and comorbidities. A team approach in treating these patients may be particularly useful. We review all current studies performed with at least one or more other modality in addition to usual medical treatment, with a focus on physical and occupational therapy. Emphasis on physical and occupational therapy with an explanation of their methods and role in multidisciplinary treatment is a pivotal part of this review. We also suggest approaches to setting up a multimodality clinic for the busy headache clinician. Setting up a collaborative, multidisciplinary team of specialists in headache practices with the goal of modifying physical, environmental, and psychological triggers for chronic daily headaches may facilitate treatment of these refractory patients.


Protein Signatures to Trace Seafood Contamination and Processing.
This review presents some applications of proteomics and selected spectroscopic methods to validate certain aspects of seafood traceability. After a general introduction to traceability and the initial applications of proteomics to authenticate traceability information, it addresses the application of proteomics to trace seafood exposure to some increasingly abundant emergent health hazards with the potential to indicate the geographic/environmental origin, such as microplastics, triclosan and human medicinal and recreational drugs. Thereafter, it shows the application of vibrational spectroscopy (Fourier-Transform Infrared Spectroscopy (FTIR) and Fourier-Transform Raman Spectroscopy (FT Raman)) and Low Field Nuclear Magnetic Resonance (LF-NMR) relaxometry to discriminate frozen fish from thawed fish and to estimate the time and temperature history of frozen fillets by monitoring protein modifications induced by processing and storage. The review concludes indicating near future trends in the application of these techniques to ensure seafood safety and traceability.


Comparison of the outcome of porcine bioprosthetic versus mechanical prosthetic replacement of the tricuspid valve in the Ebstein anomaly.
Our objective was to determine the relative merits of using a bioprosthetic porcine valve (BPV) versus a mechanical valve (MechV) when tricuspid valve (TV) replacement is required in patients with Ebstein anomaly. From 1972 to 2006, 333 patients received a BPV and 45 received a MechV. Patient records were reviewed, vital status ascertained, and all patients not known to be deceased were mailed a medical questionnaire or contacted by telephone. Early mortality was not statistically higher for patients who had a MechV (11%) than for those who had a BPV (5%) inserted in the TV position (p = 0.173). The only independent preoperative predictor of operative mortality was moderate to severe left ventricular dysfunction (odds ratio 3.1, p = 0.03); 20-year survival was better in patients who had a BPV (75%) than for those who had a MechV (43%, p = 0.003). On multivariate analysis, after adjusting for ablation of accessory pathways, sinus rhythm at dismissal, and concomitant repair of pulmonary valve stenosis, a BPV remained a predictor of late survival (hazard ratio 0.42, p = 0.004). Survival free of reoperation on the TV at 20 years postoperatively was similar for patients who had a MechV (49%) compared with those who had a BPV (42%) inserted (p = 0.941). A greater percentage of patients who had a MechV reported endocarditis (12% vs 2%), bleeding requiring hospitalization (6% vs 3%), and thrombosis (12% vs 6%); however, none of these differences were statistically significant. In conclusion, a BPV in the tricuspid position was an independent predictor of improved survival. This may be related to the higher incidence of bleeding and thrombotic complications in the patients with MechVs or may be related to differences between the 2 groups. A BPV may offer superior late survival when compared with a MechV when TV replacement is required in patients with Ebstein anomaly, but patient selection must be individualized.


Heterogeneous local dynamics revealed by classification analysis of spatially disaggregated time series data.
Time series data provide a crucial window into infectious disease dynamics, yet their utility is often limited by the spatially aggregated form in which they are presented. When working with time series data, violating the implicit assumption of homogeneous dynamics below the scale of spatial aggregation could bias inferences about underlying processes. We tested this assumption in the context of the 2015-2016 Zika epidemic in Colombia, where time series of weekly case reports were available at national, departmental, and municipal scales. First, we performed a descriptive analysis, which showed that the timing of departmental-level epidemic peaks varied by three months and that departmental-level estimates of the time-varying reproduction number, R(t), showed patterns that were distinct from a national-level estimate. Second, we applied a classification algorithm to six features of proportional cumulative incidence curves, which showed that variability in epidemic duration, the length of the epidemic tail, and consistency with a cumulative normal density curve made the greatest contributions to distinguishing groups. Third, we applied this classification algorithm to data simulated with a stochastic transmission model, which showed that group assignments were consistent with simulated differences in the basic reproduction number, R0. This result, along with associations between spatial drivers of transmission and group assignments based on observed data, suggests that the classification algorithm is capable of detecting differences in temporal patterns that are associated with differences in underlying drivers of incidence patterns. Overall, this diversity of temporal patterns at local scales underscores the value of spatially disaggregated time series data.


Prognostic impact of primary tumor resection and lymph node dissection in stage IV colorectal cancer with unresectable metastasis: a propensity score analysis in a multicenter retrospective study.
Retrospective studies have shown that primary tumor resection improves the prognosis of patients with colorectal cancer with unresectable metastasis (mCRC). Prognostic significance of lymph node dissection (LND) in mCRC has not been examined previously. The aim of this study was to investigate the prognostic impact of primary tumor resection and LND in mCRC. A total of 1,982 patients with mCRC from January 1997 to December 2007 were retrospectively studied. The impact of primary tumor resection and LND on overall survival (OS) was analyzed using Cox proportional hazards model and propensity score analysis to mitigate the selection bias. Covariates in the models for propensity scores included treatment period, institution, age, sex, carcinoembryonic antigen, tumor location, histology, depth, lymph node metastasis, lymphovascular invasion, and number of metastatic organs. In a multivariate analysis, primary tumor resection and treatment in the latter period were associated with an improved OS, and age over 70 years, female sex, lymph node metastasis, and multiple organ metastasis were associated with a decreased OS. In the propensity-matched cohort, patients treated with primary tumor resection showed a significantly better OS than those without tumor resection (median OS 13.8 vs. 6.3 months; p = 0.0001). Furthermore, among patients treated with primary tumor resection, patients treated with D3 LND showed a significantly better OS than those with less extensive LND (median OS 17.2 vs. 13.7 months; p < 0.0001). It was suggested that primary tumor resection with D3 LND improves the survival of patients with mCRC.


Drug therapy reviews: dietary fiber and fiber supplements in the therapy of gastrointestinal disorders.
Dietary fiber and fiber supplements are reviewed, with particular emphasis on their sources, composition and properties; physiological actions on gastrointestinal functions; and uses in gastrointestinal disease states (functional bowel disease, diverticular disease and other conditions). Adverse effects and contraindications, and the hypothesis of diet's effect on colon cancer also are discussed. Dietary fiber supplements may relieve symptoms of constipation, spastic colon, and diverticular disease; in the two latter disorders, colonic pressure relationships are altered. It is concluded that current evidence does not support other therapeutic uses for dietary fiber sonstituents, except possibly in patients with anal fissures and hemorrhoids, which can be helped by the passage of a softer stool.


Accessing Mole-Scanning through Community Pharmacy: A Pilot Service in Collaboration with Dermatology Specialists.
Early identification and treatment of malignant melanoma is crucial to prevent mortality. The aim of this work was to describe the uptake, profile of users and service outcomes of a mole scanning service in the community pharmacy setting in the UK. In addition, health care costs saved from the perspective of general practice were estimated. The service allowed patients to have concerning skin lesions scanned with a dermatoscopy device which were analyzed remotely by clinical dermatology specialists in order to provide recommendations for the patient. Patients were followed up to ascertain the clinical outcome. Data were analyzed for 6355 patients and 9881 scans across 50 community pharmacies. The majority of the scans required no further follow-up (n = 8763, 88.7%). Diagnosis was confirmed for 70.4% (n = 757/1118) of scans where patients were recommended to seek further medical attention. Of these, 44.3% were ultimately defined as normal (n = 335) and 6.2% as malignant melanoma (n = 47/757). An estimated 0.7% of scans taken as part of the service led to a confirmed diagnosis of malignant melanoma. This service evaluation has shown that a mole scanning service available within community pharmacies is effective at triaging patients and ultimately playing a part in identifying diagnoses of malignant melanoma.


Endothelial NADPH oxidase 4 protects ApoE-/- mice from atherosclerotic lesions.
Vascular reactive oxygen species (ROS) are known to be involved in atherosclerosis development and progression. NADPH oxidase 4 (Nox4) is a constitutively active ROS-producing enzyme that is highly expressed in the vascular endothelium. Nox4 is unique in its biology and has been implicated in vascular repair, however, the role of Nox4 in atherosclerosis is unknown. Therefore, to determine the effect of endothelial Nox4 on development of atherosclerosis, Apoe E-/- mice +/- endothelial Nox4 (ApoE-/- + EC Nox4) were fed a high cholesterol/high fat (Western) diet for 24 weeks. Significantly fewer atherosclerotic lesions were observed in the ApoE-/- + EC Nox4 mice as compared to the ApoE-/- littermates, which was most striking in the abdominal region of the aorta. In addition, markers of T cell populations were markedly different between the groups; T regulatory cell marker (FoxP3) was increased whereas T effector cell marker (T-bet) was decreased in aorta from ApoE-/- + EC Nox4 mice compared to ApoE-/- alone. We also observed decreased monokine induced by gamma interferon (MIG; CXCL9), a cytokine known to recruit and activate T cells, in plasma and tissue from ApoE-/- + EC Nox4 mice. To further investigate the link between endothelial Nox4 and MIG expression, we utilized cultured endothelial cells from our EC Nox4 transgenic mice and human cells with adenoviral overexpression of Nox4. In these cultured cells, upregulation of Nox4 attenuated endothelial cell MIG expression in response to interferon-gamma. Together these data suggest that endothelial Nox4 expression reduces MIG production and promotes a T cell distribution that favors repair over inflammation, leading to protection from atherosclerosis.


Justifications of physicians' choice of action.
To compare choice of actions and justifying reasons among the general public, GPs and oncologists. A postal questionnaire using four vignettes: a healthy patient reluctant to quit smoking and one demanding an X-ray, a pulmonary cancer patient reluctant to quit smoking and one demanding immunotherapy. Subjects A random sample of the general public (n = 1000), GPs (n = 200), and oncologists (n = 200). The respondents' choice of how the physician should act (general public)/would act (physicians), and, with reference to their choice of action, their justifying reasons. A majority answered that the physician should/would bring up the question about smoking cessation with the reluctant patient, justified by promotion of medical benefit and protection from harm. With reference to the reluctant cancer patient every second oncologist and every third GP would not mention smoking cessation, justified by respect for self-determination and enhancement of the relationship. Facing the patient demanding a new X-ray the general public was more in favour while the physicians rejected it. One-third of the general public answered that the seriously ill patient's demand for immunotherapy should be granted and referred to respect for self-determination and enhanced relationship. A majority of the physicians rejected such a demand, justified by promotion of a fair distribution of resources. The fear of infringing the patient's right to self-determination and harming the relationship is exaggerated by physicians. They are mostly expected to bring up smoking cessation with a reluctant patient. How to manage demanding patients is more controversial.


Heterogeneous binding sites for alpha-atrial natriuretic peptide in human umbilical cord and placenta.
The distributions of atrial natriuretic peptide (ANP) clearance receptors in human umbilical cord and placenta were investigated by using des[Gln18,Ser19,Gly20,Leu21,Gly22]ANP-(4-2 3) (C-ANP) as a specific ligand of this receptor to displace bound alpha-125I-labeled ANP. alpha-125I-ANP bound reversibly to umbilical venous and arterial intima and to fetal placental and maternal decidual tissues, with dissociation constants of 1.24 +/- 0.51, 0.58 +/- 0.19, 1.86 +/- 0.51, and 1.07 +/- 0.25 mM, respectively. Binding was reversed by 1 microM unlabeled alpha-ANP but not by unrelated peptides such as gastrin. The 1 microM C-ANP displaced bound alpha-125I-ANP from the intima of umbilical artery but not that of the vein. The specific reversible binding of alpha-125I-ANP to placental decidua but not to fetal placenta was also displaced by 1 microM C-ANP. Therefore high-affinity binding sites for alpha-ANP on the intima of the umbilical artery and on placental decidua differ from those on the umbilical vein and on fetal placental tissue; the binding sites of the intima of the umbilical artery and of the decidua in humans are consistent with clearance receptors for alpha-ANP.


Developing a schedule to identify social communication difficulties and autism spectrum disorder in young children with visual impairment.
Available observational tools used in the identification of social communication difficulties and diagnosis of autism spectrum disorder (ASD) rely partly on visual behaviours and therefore may not be valid in children with visual impairment. A pilot observational instrument, the Visual Impairment and Social Communication Schedule (VISS), was developed to aid in identifying social communication difficulties and ASD in young children with visual impairment affected by congenital disorders of the peripheral visual system (disorders of the globe, retina, and anterior optic nerve). The VISS was administered to 23 consecutive children (age range 1 y 9 mo-6 y 11 mo, mean 4 y 1 mo [SD 1.6]; 12 males, 11 females) with visual impairment (nine with severe and 14 with profound visual impairment). Item analysis was carried out by fit of the items to the Rasch model. Validity of the VISS was explored by comparison with the Childhood Autism Rating Scale (CARS) score, and the clinical ASD diagnosis (n=9). Correlation between the VISS and CARS total scores was highly significant (Spearman's rho=-0.89; p=0.01). Below threshold rating on the VISS (score of 35) showed good agreement with the clinical ASD diagnosis (sensitivity 89%, specificity 100%). This preliminary study shows the VISS to be a promising schedule to aid the identification of ASD in young children with visual impairment.


Mapping of lymphatic filariasis in Nepal.
BACKGROUND: Human infection with Wuchereria bancrofti causes a disabling parasitic disease known as lymphatic filariasis, which is a major public health and socio-economic problem in many parts of the world. At the onset of the study, little was known of the distribution of filariasis and its current importance as a public health problem in Nepal. METHODS: Epidemiological mapping was undertaken to determine the prevalence of infection by Wuchereria bancrofti in 37 districts of Nepal between July to December 2001. The study population above 15 years of age was selected, and the immunochromatographic test (ICT Filariasis) was used to screen for circulating filarial antigen (CFA). RESULTS: The overall prevalence of lymphatic filariasis from a 4,488-sample population was 13% and 33/37 districts were found to be endemic. On the basis of geographical data, the highest number of cases was found at altitudes between 500-700 m; however, a substantial number of infected individuals were found in the highly populated Kathmandu valley, at altitudes between 900-1,500 metres where transmission appears to take place. Prevalence rates above 20% were found in 11 districts (with the highest rate of 40%), 6-19% were found in 15 districts, and 0.1-5% were in 7 districts.Information on people's knowledge, attitudes and behaviour towards filariasis was also collected by means of a structured questionnaire, which is presented and discussed in the study. CONCLUSIONS: This is the most extensive study of lymphatic filariasis undertaken to date in Nepal. The study indicates that the prevalence of infection is far greater that was previously reported and that lymphatic filariasis should be a much higher health priority than currently given.


[Behaviour of near-infrared cerebral oximetry readings during percutaneous carotid angioplasty].
Transluminal percutaneous carotid angioplasty and stenting (CAS) carries a risk of cerebral ischemia, hemorrhage, or edema due to relative hyperemia and hemodynamic instability during and after the procedure. Noninvasive monitoring of near-infrared regional cerebral oxygen saturation (SrO2) offers an indirect way to estimate cerebral blood flow. To evaluate the behaviour of SrO2 during CAS and the usefulness of this variable for continuous monitoring of cerebral blood flow variation and neurological status. Prospective study of 25 patients scheduled for unilateral CAS under monitored anesthesia care. SrO, and other hemodynamic and clinical data were recorded. A change in SrO2 (deltaSrO2) of 15% or more in comparison with the baseline value and lasting more than 30 seconds was considered clinically significant. Neurological complications in the first 24 hours were also registered. Baseline SrO, ranged from 51% to 75%. With administration of papaverine SrO2 values increased by a mean (SD) of 5.6% (6%) (P<.05 vs baseline). They decreased during angioplasty, -2.5% (5.7%) (P<.05 vs baseline), increased after 5 minutes, and fell again at 30 minutes to a level 3% (6.54%) above baseline. Two patients showed signs of elevated intracranial pressure after the procedures and also had ASrO2 readings exceeding 15%; measures to lower arterial hypertension reduced SrO2 in these patients. High interindividual variability of absolute SrO2 values has been confirmed. SrO, fluctuates with maneuvers that change cerebral blood flow in the same way. Changes can precede the onset of other clinical signs.


g-C3N4/graphene oxide/SnFe2O4 ternary composite for the effective sunlight-driven photocatalytic degradation of methylene blue.
A broadly used dye, methylene blue (MB), adversely impacts human health and water resources, which triggers efficient methods for its elimination. Semiconductor-based heterogeneous photocatalysis is an environmentally friendly approach that effectively degrades organic pollutants. The purpose of the current work is to elucidate and validate the application of a promising g-C3N4/GO/SnFe2O4 (CGS) composite for the environmental remediation of methylene blue dye. The ternary CGS composite has been synthesized using a solvothermal approach. The fabricated composites were analyzed through FTIR, XRD, SEM/EDX, UV-VIS spectroscopy, TEM, and XPS. The photoactivity of composites and affecting parameters (pH, H2O2 dosage, composite amount, initial dye concentration, and irradiation time) were observed in sunlight illumination. The optimal conditions for photocatalytic degradation were pH = 5, photocatalyst dosage = 30 mg/100 mL, H2O2 dosage = 6 mM, and initial dye concentration (IDC) of 10 ppm employing ternary CGS composite, and MB dye was degraded effectively within 1 h. Ninety-eight percent degradation efficacy was attained by employing ternary CGS composite under the optimized conditions. Scavenging analysis suggested that •OH radicals were the key reactive oxygen species (ROS) responsible for the photodegradation of MB dye. Furthermore, the CGS nanocomposite exhibited outstanding recyclability of 84% after five consecutive runs, demonstrating its potential for use in practical applications, particularly pollutant removal.


Clinical aspects of neuropathic lysosomal storage disorders.
The purpose of this review is to describe neurological phenotypes associated with lysosomal storage diseases (LSDs), focusing on features arising from primary neuronal involvement. Clinical presentation, progression and genetic data, are discussed in detail in Part 2, the electronic material. Main features are summarized in Part 1. Insights gained from several observational studies are discussed. Prospective studies of the natural history of most neuronopathic LSDs have been hampered by the rarity of these conditions and the short survival of affected patients. Increasingly, longitudinal observations relating to neurological manifestations are being reported. Better clinical studies are necessary, including repeated measurements of disease progression to facilitate the development of sensitive scoring systems and appropriate counseling of affected individuals and their families. Ideally, clinical studies should involve a large cohort. As treatment becomes available, knowledge of disease expression and factors that influence the phenotype may enable critical assessment of therapeutic outcomes. It is hoped that increased familiarity with the clinical expression of individual LSDs will allow early diagnosis, so families at risk are given options to consider during future pregnancies. Early diagnosis also permits the introduction of timely intervention, to favoring improved outcome in cases that are potentially treatable.


The effects of disruption of phosphoglucose isomerase gene on carbon utilisation and cellulase production in Trichoderma reesei Rut-C30.
Cellulase and hemicellulase genes in the fungus Trichoderma reesei are repressed by glucose and induced by lactose. Regulation of the cellulase genes is mediated by the repressor CRE1 and the activator XYR1. T. reesei strain Rut-C30 is a hypercellulolytic mutant, obtained from the natural strain QM6a, that has a truncated version of the catabolite repressor gene, cre1. It has been previously shown that bacterial mutants lacking phosphoglucose isomerase (PGI) produce more nucleotide precursors and amino acids. PGI catalyzes the second step of glycolysis, the formation of fructose-6-P from glucose-6-P. We deleted the gene pgi1, encoding PGI, in the T. reesei strain Rut-C30 and we introduced the cre1 gene in a Δpgi1 mutant. Both Δpgi1 and cre1+Δpgi1 mutants showed a pellet-like and growth as well as morphological alterations compared with Rut-C30. None of the mutants grew in media with fructose, galactose, xylose, glycerol or lactose but they grew in media with glucose, with fructose and glucose, with galactose and fructose or with lactose and fructose. No growth was observed in media with xylose and glucose. On glucose, Δpgi1 and cre1+Δpgi1 mutants showed higher cellulase activity than Rut-C30 and QM6a, respectively. But in media with lactose, none of the mutants improved the production of the reference strains. The increase in the activity did not correlate with the expression of mRNA of the xylanase regulator gene, xyr1. Δpgi1 mutants were also affected in the extracellular β-galactosidase activity. Levels of mRNA of the glucose 6-phosphate dehydrogenase did not increase in Δpgi1 during growth on glucose. The ability to grow in media with glucose as the sole carbon source indicated that Trichoderma Δpgi1 mutants were able to use the pentose phosphate pathway. But, they did not increase the expression of gpdh. Morphological characteristics were the result of the pgi1 deletion. Deletion of pgi1 in Rut-C30 increased cellulase production, but only under repressing conditions. This increase resulted partly from the deletion itself and partly from a genetic interaction with the cre1-1 mutation. The lower cellulase activity of these mutants in media with lactose could be attributed to a reduced ability to hydrolyse this sugar but not to an effect on the expression of xyr1.


The Klebsiella pneumoniae cytochrome bd' terminal oxidase complex and its role in microaerobic nitrogen fixation.
Cytochrome bd' has been implicated in having an important role in microaerobic nitrogen fixation in the enteric bacterium Klebsiella pneumoniae, where it is expressed under all conditions that permit diazotrophy. In this paper the sequence of the genes encoding this terminal oxidase (cydAB) of Klebsiella pneumoniae and the characterization of a cyd mutant are reported. The deduced amino acid sequences support the proposal that His 19, His 186 and Met 393 provide three of the four axial ligands to the Fe of the three haems in the oxidase complex. The nitrogen-fixing ability of the mutant was severely impaired in the presence of low concentrations of oxygen compared with the wild-type bacterium. Only the wild-type organism was capable of microaerobic nitrogenase activity supported by fermentation products. It is proposed that formate dehydrogenase-O may be involved in supplying electrons to a respiratory chain terminated by the bd-type oxidase, which would remove inhibitory oxygen and supply ATP for nitrogenase activity.


A detailed fluid mechanics study of tilting disk mechanical heart valve closure and the implications to blood damage.
Hemolysis and thrombosis are among the most detrimental effects associated with mechanical heart valves. The strength and structure of the flows generated by the closure of mechanical heart valves can be correlated with the extent of blood damage. In this in vitro study, a tilting disk mechanical heart valve has been modified to measure the flow created within the valve housing during the closing phase. This is the first study to focus on the region just upstream of the mitral valve occluder during this part of the cardiac cycle, where cavitation is known to occur and blood damage is most severe. Closure of the tilting disk valve was studied in a "single shot" chamber driven by a pneumatic pump. Laser Doppler velocimetry was used to measure all three velocity components over a 30 ms period encompassing the initial valve impact and rebound. An acrylic window placed in the housing enabled us to make flow measurements as close as 200 microm away from the closed occluder. Velocity profiles reveal the development of an atrial vortex on the major orifice side of the valve shed off the tip of the leaflet. The vortex strength makes this region susceptible to cavitation. Mean and maximum axial velocities as high as 7 ms and 20 ms were recorded, respectively. At closure, peak wall shear rates of 80,000 s(-1) were calculated close to the valve tip. The region of the flow examined here has been identified as a likely location of hemolysis and thrombosis in tilting disk valves. The results of this first comprehensive study measuring the flow within the housing of a tilting disk valve may be helpful in minimizing the extent of blood damage through the combined efforts of experimental and computational fluid dynamics to improve mechanical heart valve designs.


Derivation and validation of a MEDLINE search strategy for research studies that use administrative data.
To derive and validate a search strategy that identifies administrative database research (ADR) in the MEDLINE database. Analytical survey. We downloaded all articles published between January 1, 2008 and October 7, 2009 in 20 top journals in internal medicine, cardiovascular medicine, public health, and health services research. These were reviewed to determine whether they were ADR (in which the study cohort, exposure, or outcome was defined using electronic data created for or during the processing of patients through their health care). We used chi-squared recursive partitioning to create a search strategy that maximized sensitivity based on publication type, MeSH headings, and text words. Sensitivity and positive predictive value of the search strategy for true ADR in three samples: derivation (n=5,513); internal validation (n=2,710); and external validation (n=1,500). The prevalence of ADR in the derivation, internal validation, and external validation samples was 2.6, 2.9, and 2.2 percent, respectively. The sensitivity of our search strategy in these samples was 90.9 percent (95 percent confidence interval [CI] 85.0-95.1), 88.5 percent (79.2-94.6), and 100 percent (99.3-100), respectively. The positive predictive value in these samples was 10.7 percent (9.0-12.6), 11.5 percent (9.1-14.4), and 3.3 percent (2.3-4.6), respectively. We derived and validated a search strategy that is highly sensitive for ADR in MEDLINE.


A specific population of gonadotrophs purified from immature female rat pituitary.
When dispersed pituitary cells from 14-day-old female rats were sedimented in a bovine serum albumin gradient, a fraction was isolated which consisted of almost 90 percent of large cells that stained purple in the periodic acid-Schiff (PAS) reaction. Immunostaining indicated that over 85 percent of these PAS-purple cells were gonadotrophs containing both follicle stimulating and luteinizing hormone. Reproducible cell cultures could be obtained on poly-L-lysine coated dishes. As early as the second day in culture, the secretion of both follicle stimulating and luteinizing hormone was highly stimulated by luteinizing hormone-releasing hormone. The effect of FSH was as marked as that on LH. The data suggest that the isolated gonadotrophs are a specific functional subtype.


Antineutrophil Cytoplasmic Antibody-associated Vasculitis Superimposed on Infection-related Glomerulonephritis Secondary to Pulmonary Mycobacterium avium Complex Infection.
A 73-year-old woman was diagnosed with pulmonary Mycobacterium avium complex (MAC) infection and received no treatment. Disease progression was evident one year later with the development of myeloperoxidase-antineutrophil cytoplasmic antibody (ANCA) titers and systemic symptoms of a fever, polyarthritis, purpura, and rapidly progressive glomerulonephritis. Her symptoms did not improve with antibiotic treatment. A renal biopsy revealed crescentic glomerulonephritis with immunodeposition. According to these findings, she was diagnosed with ANCA-associated vasculitis (AAV) superimposed on infection-related glomerulonephritis (IRGN). Although there was a risk of aggravating an underlying infection, the combination therapy of corticosteroid and antibiotics improved AAV, IRGN, and even the lung radiological findings. To the best of our knowledge, this is the first case of AAV and IRGN secondary to pulmonary MAC infection.


Sensory Effects of Nicotine and Tobacco.
Ingestion of nicotine by smoking, vaping, or other means elicits various effects including reward, antinociception, and aversion due to irritation, bitter taste, and unpleasant side effects such as nausea and dizziness. Here we review the sensory effects of nicotine and the underlying neurobiological processes. Nicotine elicits oral irritation and pain via the activation of neuronal nicotinic acetylcholine receptors (nAChRs) expressed by trigeminal nociceptors. These nociceptors excite neurons in the trigeminal subnucleus caudalis (Vc) and other brainstem regions in a manner that is significantly reduced by the nAChR antagonist mecamylamine. Vc neurons are excited by lingual application of nicotine and exhibit a progressive decline in firing to subsequent applications, consistent with desensitization of peripheral sensory neurons and progressively declining ratings of oral irritation in human psychophysical experiments. Nicotine also elicits a nAChR-mediated bitter taste via excitation of gustatory afferents. Nicotine solutions are avoided even when sweeteners are added. Studies employing oral self-administration have yielded mixed results: Some studies show avoidance of nicotine while others report increased nicotine intake over time, particularly in adolescents and females. Nicotine is consistently reported to increase human pain threshold and tolerance levels. In animal studies, nicotine is antinociceptive when delivered by inhalation of tobacco smoke or systemic infusion, intrathecally, and by intracranial microinjection in the pedunculopontine tegmentum, ventrolateral periaqueductal gray, and rostral ventromedial medulla. The antinociception is thought to be mediated by descending inhibition of spinal nociceptive transmission. Menthol cross-desensitizes nicotine-evoked oral irritation, reducing harshness that may account for its popularity as a flavor additive to tobacco products. Nicotine activates brain systems underlying reward and antinociception, but at the same time elicits aversive sensory effects including oral irritation and pain, bitter taste, and other unpleasant side effects mediated largely by nicotinic acetylcholine receptors (nAChRs). This review discusses the competing aversive and antinociceptive effects of nicotine and exposure to tobacco smoke, and the underlying neurobiology. An improved understanding of the interacting effects of nicotine will hopefully inform novel approaches to mitigate nicotine and tobacco use.


Ultrasound Elasticity Imaging Predicts Therapeutic Outcomes of Patients With Crohn's Disease Treated With Anti-Tumour Necrosis Factor Antibodies.
Ultrasound elasticity imaging is a non-invasive technique developed to evaluate fibrosis. Measuring tissue strain by ultrasound elasticity imaging can reliably detect severe ileal fibrosis in patients with Crohn's disease [CD]. We have hypothesised that a more severe range of fibrosis might influence the therapeutic response to anti-tumour necrosis factor [TNF] treatment. The aim of this study was to assess the ability of ultrasound elasticity imaging to predict the therapeutic outcome for CD patients. Consecutive patients with ileal/ileocolonic CD, starting anti-TNF treatment, were enrolled for the study. These patients underwent bowel ultrasound and ultrasound elasticity imaging at baseline and at 14 and 52 weeks after anti-TNF treatment. Bowel wall stiffness was quantified by calculating the strain ratio between the mesenteric tissue and the bowel wall. Strain ratio ≥ 2 was used to identify severe ileal fibrosis. Transmural healing at 14 and 52 weeks was defined as bowel wall thickness ≤ 3 mm. Thirty patients with CD were enrolled. Five patients underwent surgery for bowel obstruction. The frequency of surgeries was significantly greater in patients with a strain ratio ≥ 2 at baseline [p = 0.003]. A significant reduction of the bowel thickness was observed after 14 and 52 weeks of anti-TNF treatment [p < 0.005]. A significant inverse correlation was observed between the strain ratio values at baseline and the thickness variations following anti-TNF therapy [p = 0.007]; 27% of patients achieved transmural healing at 14 weeks. The baseline strain ratio was significantly lower in patients with transmural healing [p < 0.05]. This study shows that ultrasound elasticity imaging predicts therapeutic outcomes for CD patients treated with anti-TNF.


Multiscale simulations reveal the role of PcrA helicase in protecting against spontaneous point mutations in DNA.
Proton transfer across hydrogen bonds in DNA can produce non-canonical nucleobase dimers and is a possible source of single-point mutations when these forms mismatch under replication. Previous computational studies have revealed this process to be energetically feasible for the guanine-cytosine (GC) base pair, but the tautomeric product (G[Formula: see text]C[Formula: see text]) is short-lived. In this work we reveal, for the first time, the direct effect of the replisome enzymes on proton transfer, rectifying the shortcomings of existing models. Multi-scale quantum mechanical/molecular dynamics (QM/MM) simulations reveal the effect of the bacterial PcrA Helicase on the double proton transfer in the GC base pair. It is shown that the local protein environment drastically increases the activation and reaction energies for the double proton transfer, modifying the tautomeric equilibrium. We propose a regime in which the proton transfer is dominated by tunnelling, taking place instantaneously and without atomic rearrangement of the local environment. In this paradigm, we can reconcile the metastable nature of the tautomer and show that ensemble averaging methods obscure detail in the reaction profile. Our results highlight the importance of explicit environmental models and suggest that asparagine N624 serves a secondary function of reducing spontaneous mutations in PcrA Helicase.


Maladaptive plasticity: imprinting of past experiences onto phantom limb schemata.
Phantom limb perception is common following amputation, and is sometimes characterised by pain that resembles the characteristics, intensity or location of past pain. We tested Flor's model that phantom pain results from memory for long-lasting znoxious input. We report a questionnaire study of 283 amputees, that explored the experience of painful, non-painful and postural somatosensory memories in the phantom. We explore the impact of pre-amputation pain and impairment duration, and complications in the limb (eg, infection, gangrene, surgery, and vascular disease). Differences in mood, coping and adjustment to amputation are also explored in those with somatosensory pain memories. Our findings support Flor's model, as amputation-related and non-amputation-related pain memories, and non-painful memories comprised pains or sensations that were either enduring/recurring pains or sensations (eg, ingrown toenail, corns, chilblains, arthritis-type pain in winter, night-cramps, or holding a tennis racquet), or resulted from a painful event with a "core-trauma" element (eg, fracture, crushing/penetration injury). Pain memories related to amputation were more common following functional impairment before amputation; infection or surgery prior to amputation; or having diabetic or vascular amputations-which are associated with multiple complications, including neuropathic changes, infection and prior surgery. Furthermore, participants with amputation-related pain memories exhibited higher sensory pain ratings, as well as poorer mood and adjustment to the limitations of amputation. We propose that somatosensory pain memories likely relate to the generation and maintenance of limb representations upon which intense or emotionally powerful past experiences have been imprinted.


Contribution of socioeconomic factors and health care access to the awareness and treatment of diabetes and hypertension among older Mexican adults.
To estimate changes in self-report and treatment of diabetes and hypertension between 2001 and 2012 among Mexican aged 50-80, assessing the contribution of education and health insurance coverage. The Mexican Health and Aging Study was used to estimate associations of education and insurance on prevalence and treatment of diabetes and hypertension in 2001 and 2012. Multivariate decomposition was used to assess the contribution of changes in the composition of covariates vs. their "effects" on changes in prevalence and treatment over time. Increases in the prevalence/diagnosis and treatment during the period are largely attributable to the expansion of health insurance. Its effects on diagnosis/prevalence and treatment have also increased over time. The expansion of Seguro Popular likely improved screening and treatment. More research is needed to assess if these have translated into better control and a lower burden of disease.


Electron back-scattering diffraction (EBSD) measurements of antigorite lattice-preferred orientations (LPO).
Lattice preferred orientations of serpentines induce a strong anisotropy of various properties in serpentine bearing-rocks. Lattice preferred orientations had so far been obtained only by X-ray diffraction techniques. We have applied electron back-scattering diffraction to the measurement of the lattice preferred orientations of antigorite in a naturally deformed high-pressure serpentinite. This technique is very sensitive to sample preparation that can lead to surface amorphization in the case of serpentine. A polishing procedure is described that avoids amorphization and allows accurate electron back-scattering diffraction measurements with optimized experimental conditions in a variable pressure scanning electron microscope. Results indicate that deformation leads to lattice preferred orientations characterized by extremely strong c-axis clustering perpendicular to the foliation, as expected for a layered silicate. In the foliation plane, a significant clustering of the a-axis is observed and tentatively attributed to intracrystalline deformation mechanisms. These data suggest that antigorite deforms mostly by gliding along the basal plane of the layered phyllosilicate structure, but that gliding may occur along directions favouring a-axis alignment. Electron back-scattering diffraction appears to be a reliable method for determining phyllosilicate lattice preferred orientations in deformed rocks, with potential applications for determining anisotropy of properties like seismic velocities or thermal and electrical conductivities.