Archaeal signal transduction: impact of protein phosphatase deletions on cell size, motility, and energy metabolism in Sulfolobus acidocaldarius.
In this study, the in vitro and in vivo functions of the only two identified protein phosphatases, Saci-PTP and Saci-PP2A, in the crenarchaeal model organism Sulfolobus acidocaldarius were investigated. Biochemical characterization revealed that Saci-PTP is a dual-specific phosphatase (against pSer/pThr and pTyr), whereas Saci-PP2A exhibited specific pSer/pThr activity and inhibition by okadaic acid. Deletion of saci_pp2a resulted in pronounced alterations in growth, cell shape and cell size, which could be partially complemented. Transcriptome analysis of the three strains (Δsaci_ptp, Δsaci_pp2a and the MW001 parental strain) revealed 155 genes that were differentially expressed in the deletion mutants, and showed significant changes in expression of genes encoding the archaella (archaeal motility structure), components of the respiratory chain and transcriptional regulators. Phosphoproteome studies revealed 801 unique phosphoproteins in total, with an increase in identified phosphopeptides in the deletion mutants. Proteins from most functional categories were affected by phosphorylation, including components of the motility system, the respiratory chain, and regulatory proteins. In the saci_pp2a deletion mutant the up-regulation at the transcript level, as well as the observed phosphorylation pattern, resembled starvation stress responses. Hypermotility was also observed in the saci_pp2a deletion mutant. The results highlight the importance of protein phosphorylation in regulating essential cellular processes in the crenarchaeon S. acidocaldarius.


TcpC protein from E. coli Nissle improves epithelial barrier function involving PKCζ and ERK1/2 signaling in HT-29/B6 cells.
The probiotic Escherichia coli Nissle 1917 (EcN) is widely used to maintain remission in ulcerative colitis. This is thought to be mediated by various immunomodulatory and barrier-stabilizing effects in the intestine. In this study, the mechanisms of barrier modulation by EcN were studied in the human epithelial HT-29/B6 cell culture model.EcN supernatant increased transepithelial resistance (TER) and reduced permeability to mannitol because of sealing of the paracellular passage pathway as revealed by two-path impedance spectroscopy. This increase in TER was attributed to the TcpC protein of EcN. TcpC induced protein kinase C-ζ (PKCζ) and extracellular-signal-regulated kinase 1/2 (ERK1/2) phosphorylation, which in turn resulted in upregulation of the barrier-forming tight junction protein claudin-14. By specific silencing of protein expression by small interfering RNA (siRNA), the sealing function of claudin-14 was confirmed. In conclusion, the TcpC protein of EcN affects innate immunity by improving intestinal barrier function through upregulation of claudin-14 via PKCζ and ERK1/2 signaling.


Drug-Drug Interactions Among Hospitalized Children Receiving Chronic Antiepileptic Drug Therapy.
Children treated with chronic medications are at risk of drug-drug interactions (DDIs) when hospitalized with an acute illness and prescribed new medications. We aimed to measure the prevalence of potential DDIs (pDDIs) among hospitalized children treated with antiepileptic drugs (AEDs) and to evaluate the impact of computerized physician order entry (CPOE) on pDDIs. We analyzed a national sample of pediatric hospitalizations from 2005 to 2012 associated with administration of an AED and identified those prescribed a second medication with risk of a DDI. The prevalence of hospitalizations associated with a pDDI was calculated for each AED. We identified the drugs most commonly implicated in pDDIs and factors associated with pDDIs. Rates of pDDIs were measured in pre- and post-CPOE implementation periods. A pDDI was identified in 181 380 (41.7%) hospitalizations associated with the use of an AED, with 117 880 (27.1%) classified as severe pDDIs. AEDs most often implicated with a pDDI were phenobarbital, valproic acid, and phenytoin. Hospitalizations with pDDIs were associated with increased length of stay and a greater number of medications, ICU admissions, and operating room procedures. The implementation of CPOE did not result in a change in the rate of pDDIs (42.7% before versus 40.8% after; P = .48). Children treated with AEDs are at risk of pDDIs while hospitalized. The use of CPOE has not been associated with a significant decrease in the rate of pDDIs. Additional investigation to better define the impact of pDDIs and to advance development of clinical decision support within CPOE systems is warranted.


Core outcome measures for research in traumatic injury survivors: The National Trauma Research Action Plan modified Delphi consensus study.
Until recently, survival has been the main outcome measure for injury research. Given the impact of injury on quality of life, the National Academies of Science, Engineering, and Medicine has called for advancing the science of research evaluating the long-term outcomes of trauma survivors. This is necessary so that treatments and interventions can be assessed for their impact on a trauma patients' long-term functional and psychosocial outcomes. We sought to propose a set of core domains and measurement instruments that are best suited to evaluate long-term outcomes after traumatic injury with a goal for these measures to be adopted as a national standard. As part of the development of a National Trauma Research Action Plan, we conducted a two-stage, five-round modified online Delphi consensus process with a diverse panel of 50 key stakeholders including clinicians, researchers, and trauma survivors from more than 9 professional areas across the United States. Before voting, panelists reviewed the results of a scoping review on patient-reported outcomes after injury and standardized information on measurement instruments following the Consensus-based Standards for the Selection of Health Measurement Instruments guidelines. The panel considered a preliminary list of 74 outcome domains (patient-reported outcomes) and ultimately reached the a priori consensus criteria for 29 core domains that encompass aspects of physical, mental, social, and cognitive health. Among these 29 core domains, the panel considered a preliminary list of 199 patient-reported outcome measures and reached the a priori consensus criteria for 14 measures across 13 core domains. Participation of panelists ranged from 65% to 98% across the five Delphi rounds. We developed a core outcome measurement set that will facilitate the synthesis, comparison, and interpretation of long-term trauma outcomes research. These measures should be prioritized in all future studies in which researchers elect to evaluate long-term outcomes of traumatic injury survivors. Diagnostic Test or Criteria, Level IV.


[Blepharoplasty: various technics in surgery of eyelid aging].
The authors reviewing their own experience of blepharoplasty (92 cases) over the last ten years, discuss the principal approaches used depending on the age of the patient, the amount of adipose tissue, the lower lid tarsal and muscular laxity and the quality of the skin excess. In the lower eyelid they emphasize the value of the transconjunctival approach in young and or coloured people and the use of a cutaneo-muscular flap in the classical technique. In the upper eyelid, they combine the skin excision with an orbicularis muscle strip and in old people with suspension of the lateral canthus. A wise selection of these various approaches avoids the rare complications (ectropion and scleral show) because blepharoplasty must be considered to be a "variable geometry" operation.


Resistance and tolerance to herbivory in Solidago altissima (Asteraceae): genetic variability, costs, and selection for multiple traits.
Quantifying the genetic variability, fitness costs, and selection gradients associated with plant defense traits is necessary to understand their evolution and continued persistence in populations. Few studies have simultaneously examined the costs, benefits, and genetic variability in multiple traits related to plant resistance and tolerance to herbivory. • Using 103 Solidago altissima (Asteraceae) genets from two populations previously studied in situ, we conducted a common garden experiment to assess genetic variability, costs, selection gradients, and correlations among resistance, tolerance, and various resistance and tolerance traits (i.e., lateral branching, relative growth rate, leaf addition and senescence rate, specific leaf area, and leaf toughness). • We report evidence for significant genetic variability in resistance and various tolerance-related traits but low broad-sense heritability (H(2) < 0.14) for all traits. For all traits examined, no correlation existed between trait levels of parent ramets (measured in their field of origin) and daughter ramets (measured in the common garden), suggesting plasticity in goldenrod traits. We found a strong cost of resistance and selection gradient against high resistance. Conversely, we found no evidence of costs but did find significant selection gradients favoring increased tolerance and many tolerance trait levels. • Our study suggests that herbivores impose selection favoring increased tolerance and reduced resistance in goldenrods. In this environment, we expect that over time, resistant genets will decrease in frequency. Despite strong selection pressures, the evolution of tolerance in this environment may be constrained by the low broad-sense heritability in tolerance traits.


Acute hemoperitoneum in horses: a review of 19 cases (1992-2003).
The medical records of 19 horses with acute hemoperitoneum were reviewed. The causes for the hemoperitoneum were idiopathic (8 horses), splenic hematoma with capsular tear (7), bleeding from the reproductive tract (3), multicentric hemangiosarcoma (1), and systemic amyloidosis (1). The affected horses were between 4 and 32 years of age (median 11.5 years). The most consistent findings on initial examination were depression, tachycardia, tachypnea, pale mucous membranes, prolonged capillary refill time, colic, and abdominal discomfort. Less common clinical signs included abdominal distention, profuse sweating, ataxia, and broad ligament mass palpated on rectal examination. Clinicopathologic abnormalities commonly detected were anemia, neutrophilia, lymphopenia, thrombocytopenia, hypoproteinemia, hypocalcemia, azotemia, increased creatinine kinase, and sorbitol dehydrogenase activity. Hemoperitoneum was diagnosed on the basis of abdominocentesis, transabdominal ultrasonography, and postmortem examination. Sixteen horses were treated, and 3 horses were euthanized at owners' request because of severe clinical signs. The treatment consisted of the administration of intravenous fluids, plasma or blood transfusion, nonsteroidal drugs, antimicrobial drugs, and antifibrinolytic and procoagulant agents. Rapid clinical deterioration was observed in 2 horses, necessitating euthanasia. The remaining 14 horses survived the abdominal bleeding (survival rate 74%) and were discharged 3-15 days (median 7.0 days) after presentation. Postmortem examination of the 6 nonsurvivors showed massive abdominal hemorrhage from splenic hematoma with capsular tear (2 horses), multicentric hemangiosarcoma with liver rupture (1), systemic amyloidosis with splenic hematoma and capsular tear (1), and bilateral ruptured ovarian hematomas (1). In one horse, no origin of the bleeding could be determined during postmortem examination.


Bactericidal mechanisms revealed for rapid water disinfection by superabsorbent cryogels decorated with silver nanoparticles.
The authors have recently reported the fabrication of superabsorbent cryogels decorated with silver nanoparticles (PSA/AgNP cryogels) that demonstrate rapid water disinfection. This paper provides a systematic elucidation of the bactericidal mechanisms of AgNPs (silver nanoparticles), both generally and in the specific context of cryogels. Direct contact between the PSA/AgNP cryogel interface and the bacterial cells is required to accomplish disinfection. Specifically, the disinfection efficacy is closely correlated to the cell-bound Ag concentration, which constitutes >90% of the Ag released. Cells exposed to PSA/AgNP cryogels show a significant depletion of intracellular adenosine triphosphate (ATP) content and cell-membrane lesions. A positive ROS (reactive oxygen species) scavenging test confirms the involvement of ROS (·O2(-), H2O2, and ·OH) in the bactericidal mechanism. Furthermore, exposed bacterial cells show an enhanced level of thiobarbituric acid reactive substances, indicating the occurrence of cell-membrane peroxidation mediated by ROS. In addition, this study reveals that both Ag(+) and Ag(0) are involved in the bactericidal mechanism of AgNPs via tests conducted using PSA cryogels with bound Ag(+) ions (or PSA/Ag(+) cryogels without reducing Ag(+) to Ag(0)). Significantly, bacterial cells exposed to PSA/Ag(+) cryogels did not show any cell-membrane damage even though the former had a higher cell-bound Ag concentration than that of the PSA/AgNP cryogels, thus indicating the differential action of Ag(+) and Ag(0).


Qualitative identification of rigid gas permeable contact lens materials by densitometry.
We describe a practical method to qualitatively identify polymethylmethacrylate (PMMA) and rigid gas permeable (RGP) contact lens materials. By progressive dilution of a saturated saline solution made with distilled or tap water and sodium chloride, we recorded comparative densitometry of rigid contact lens materials using a small hydrometer or by liquid displacement. The method was sensitive enough to separate the polymethylmethacrylate, all silicon-methacrylates, and all but two fluorine-containing silicon-methacrylates. The hydrometer had a precision of three decimals rounded to the nearest 0.005. There was only one RGP product that could have been confused with the PMMA material. Most silicon-methacrylates had lower densities than fluorine containing silicon-methacrylates. Only four of 25 products under 1.117 gm/cm3 contained fluorine. Densitometry with a hydrometer is an effective non-destructive method to identify RGP materials and to verify their quality. The method is easier when lens blanks are tested, but in spite of differences in shape, size, and weight, densitometry may also be used with new or used contact lenses. Its simplicity and low cost makes densitometry feasible for any contact lens laboratory or clinic to use on a routine basis. Only silicon-methacrylates had an inverse relationship between density and oxygen permeability. As the silicon content of the contact lens increases, the Dk increases and the density decreases.


Rationale and design of the Turkish acute myocardial infarction registry: The TURKMI Study.
To the best of our knowledge, there is no up-to-date information regarding the presentation, management, and clinical course of patients with acute myocardial infarction (MI) in Turkey. The TURKMI registry is designed to provide an insight into the characteristics, management from the symptoms onset to the hospital discharge, and outcome of patients with acute MI in Turkey. The TURKMI study, as a nationwide registry, will be conducted in 50 percutaneous coronary intervention-capable centers, selected from each EuroStat NUTS region in Turkey according to their population sampling weight, prioritizing the hospital volume in each region. All consecutive patients with acute MI admitted to the coronary care units within the 48 hours of the symptoms onset will be prospectively enrolled during a predefined 2-week period. The first step of the study has a cross-sectional design in which baseline information such as symptoms, risk factors, time periods at each step from the symptoms onset to revascularization, way of arrival to hospital, biochemical analysis, and in-hospital management and outcome will be assessed. The second step has a cohort characteristic in which the enrolled patients will be followed-up up to 2 years. Follow-up visits will be conducted at the 1st, 6th, 12th, and 24th month, and predictors and risk of cardiovascular events and implementation of guidelines will be assessed as secondary outcomes. The national TURKMI registry is expected to provide important information to improve the national policy regarding diagnosing, management, and outcomes of MI in Turkey.


[Participation of bone-marrow stem cells in the differentiation of mdx mice striated muscle].
Two sets of experiments were carried out. The first one involved chimeric mice, obtained by intravenously injections of bone marrow derived cells taken from transgenic C57BL/6 mice, expressing GFP, to 5 Gy X-ray irradiated mdx or C57BL/6 mice. In 2 months M. quadriceps femoris of chimeric mice were destroyed by surgical clamp. Following the next 4-5 weeks, the same muscles were studied for the presence of GFP-positive striated muscle fibres. In the case of chimeric C57BL/6 mice GFP-positive striated muscle fibres were observed in 0.3 +/- 0.5 and in 0.2 +/- 0.3 % of destroyed muscle, and in lateral (control) muscle, consequently. In the case of chimeric mdx mice, positive results were observed in 1.7 +/- 0.4 and in 0.5 +/- 0.3 % of destroyed and control muscles, respectively. In the second set of experiments, the GFP-positive bone marrow cells were used for multiple intramuscular injections to M. quadriceps femoris of C57BL/6 or mdx mice in a dose of 2 x 10(5)-5 x 10(5) cells per mouse. Before injection, GFP-positive bone marrow cells were fractionated in a 63 % Percoll solution and then were exhausted from differentiated cells by magnetic manner using CD4, CD8, CD38, CD45R, CD119, Ly-6G, and F4/80 antibodies. After 2-3 weeks, as many as 0.15 +/- 0.40 and 0.1 +/- 0.2 % of GFP-positive muscle fibres were found in injected and control muscles of C57BL/6 mice, respectively. In the case of mdx mice, the frequency of GFP-positive striated muscle fibres was 2.0 +/- 0.8 and 1.2 +/- 0.6 % for injected and control muscles, respectively. A conclusion is made that bone marrow stem cells can take part in differentiation of mdx mouse muscles after their delivery by needle injections.


Intermittent Horner's syndrome: recurrent, alternate Horner's syndrome in cervical cord injury.
Ten patients with functional transection of the lower cervical cord were noted to develop a transient, occasionally alternate Horner syndrome. The ocular signs were rarely complete, eyelid droop being the most constant and prominent symptom. Signs could be elicited by turning the patients from side to side. They disappeared gradually with patients lying recumbent or supine. Cocaine and Adrenalin in the presently used concentration are not useful in diagnosis, but cocaine may possibly be used to "bring out" a "latent Horner's syndrome." The appearance of eye symptoms is delayed from 2 months to 7 years and, although implicating cavitation within the central gray, is not necessarily associated with progression of cord destruction rostrally or loss of remaining function. Temporary interference with sympathetic stimulation along the lateral tectotegmental tracts discharging to the intermediolateralis sympathetic column supplying the eye is suggested as responsible for this phenomenon. Fluid withdrawal from these cysts has been reported to revert the eye findings to normal and possibly arrest further cavitation.


Dermoscopic features of clear cell acanthoma.
Clear cell acanthoma is an uncommon type of benign epithelial tumor. Typically, it is a solitary lesion found on the lower limbs. It presents as a nodule or small plaque with slow and well-defined growth. Diagnosis used to be clinical and histopathological, but the advent of dermoscopy has led to an increase in diagnostic accuracy. We describe a case in which dermoscopy proved highly useful for diagnosis of the lesion.


The effects of low-volume resistance training with and without advanced techniques in trained subjects.
This study examined low-volume resistance training (RT) in trained participants with and without advanced training methods. Trained participants (RT experience 4±3 years) were randomised to groups performing single-set RT: ssRM (N.=21) performing repetitions to self-determined repetition maximum (RM), ssMMF (N.=30) performing repetitions to momentary muscular failure (MMF), and ssRP (N.=28) performing repetitions to self-determined RM using a rest pause (RP) method. Each performed supervised RT twice/week for 10 weeks. Outcomes included maximal isometric strength and body composition using bioelectrical impedance analysis. The ssRM group did not significantly improve in any outcome. The ssMMF and ssRP groups both significantly improved strength (p < 0.05). Magnitude of changes using effect size (ES) was examined between groups. Strength ES's were considered large for ssMMF (0.91 to 1.57) and ranging small to large for ssRP (0.42 to 1.06). Body composition data revealed significant improvements (P<0.05) in muscle and fat mass and percentages for whole body, upper limbs and trunk for ssMMF, but only upper limbs for ssRP. Body composition ES's ranged moderate to large for ssMMF (0.56 to 1.27) and ranged small to moderate for ssRP (0.28 to 0.52). ssMMF also significantly improved (P<0.05) total abdominal fat and increased intracellular water with moderate ES's (-0.62 and 0.56, respectively). Training to self-determined RM is not efficacious for trained participants. Training to MMF produces greatest improvements in strength and body composition, however, RP style training does offer some benefit.


[Distribution of therapeutic procedures and choice of drug therapies in patients with angiographically confirmed coronary artery disease].
The aims of our study were to assess the distribution of interventional and other therapeutic procedures performed on subjects who had proven CAD by angiography in our clinic; to determine the groups of therapeutic agents prescribed at the time of discharge; and to compare these with the results of EUROASPIRE II, which examined the prophylactic drug therapy upon discharge of CAD patients in 15 European countries. Our patients comprises of 617 subjects (516 male, 101 female; mean age 57.2 +/- 10.8 years) who underwent coronary angiography for the first time in our clinic and who were found to have a 50% lesion in at least one of the coronary arteries. In all patients distribution of risk factors on admission, distribution of therapeutic procedures and the use of drug therapies at the hospital discharge were recorded. We found that, 68% of our cases were considered to be eligible for a percutaneous or surgical intervention, while 27% were assessed not as requiring such an intervention and consequently were discharged being prescribed appropriate medications. For the remaining 5% of the subjects, tests for detection of viable myocardium were advised, before selecting the proper type of management. At discharge, prescription rates for antiplatelets, beta-blockers, nitrates, statins, angiotensin-converting enzyme inhibitors (ACE-I), calcium channel blockers and anticoagulants were detected to be %99, 86%, 86%, 63%, 40%, 16%, and 2% respectively. Compared with the results of EUROASPIRE II study, these data show that, antiplatelet, beta-blocker, ACE-I and lipid-lowering treatments our subjects received upon discharge were higher, whereas anticoagulant treatment was lower than the averages of the fifteen European countries consisting the EUROASPIRE II study.


Efficient in utero gene transfer system to the developing mouse brain using electroporation: visualization of neuronal migration in the developing cortex.
We report a novel gene transfer system using electroporation. We used this technique to introduce a marker gene plasmid containing enhanced green fluorescent protein into mouse brains at embryonic day 12-17 without removing the embryos from the uterus. The embryos were allowed to continue to develop in utero, and more than 80% were born normally expressing the exogenous gene. Enhanced green fluorescent protein driven by the cytomegalovirus promoter was strongly expressed in the ventricular zone, radial fibers and migrating neuroblasts, but not in mature neurons, suggesting that the cytomegalovirus promoter is silenced after the cells differentiate into mature neurons. Since there is still no convenient way of visualizing the migrating neuroblasts, especially of distinguishing them from the surrounding mature neurons in the cortical plate, this system should provide a good tool for analysing neuronal migration. In the postnatal lateral cortex, neuroblasts migrated almost "tangentially" along the obliquely running "radial" fibers beneath the cortical plate, and after entering the cortical plate, turned towards the marginal zone and migrated radially. Neurons with primitive dendrites were observed only along the border between the marginal zone and the cortical plate, and never at other sites, such as in the middle of the cortical plate. These results imply that the neuroblasts do terminate migration and start differentiation to mature neurons when they encounter the marginal zone, as has long been suggested. By contrast, when elongation factor 1alpha promoter was used, prominent fluorescence allowed visualization of the entire mature neurons as well. The labeled neurons were observed to send axons to the contralateral cortex where they arborized extensively.Thus, this system is much easier and more efficient than virus-mediated gene transfer, and is useful for gain-of-function analysis of neural cell fate determination, migration, positioning and axon path-finding in mouse embryos.


Combination of fluorescence microscopy and nanomotion detection to characterize bacteria.
Antibiotic-resistant pathogens are a major health concern in everyday clinical practice. Because their detection by conventional microbial techniques requires minimally 24 h, some of us have recently introduced a nanomechanical sensor, which can reveal motion at the nanoscale. By monitoring the fluctuations of the sensor, this technique can evidence the presence of bacteria and their susceptibility to antibiotics in less than 1 h. Their amplitude correlates to the metabolism of the bacteria and is a powerful tool to characterize these microorganisms at low densities. This technique is new and calls for an effort to optimize its protocol and determine its limits. Indeed, many questions remain unanswered, such as the detection limits or the correlation between the bacterial distribution on the sensor and the detection's output. In this work, we couple fluorescence microscopy to the nanomotion investigation to determine the optimal experimental protocols and to highlight the effect of the different bacterial distributions on the sensor.


A Predictive Model for Yeast Cell Polarization in Pheromone Gradients.
Budding yeast cells exist in two mating types, a and α, which use peptide pheromones to communicate with each other during mating. Mating depends on the ability of cells to polarize up pheromone gradients, but cells also respond to spatially uniform fields of pheromone by polarizing along a single axis. We used quantitative measurements of the response of a cells to α-factor to produce a predictive model of yeast polarization towards a pheromone gradient. We found that cells make a sharp transition between budding cycles and mating induced polarization and that they detect pheromone gradients accurately only over a narrow range of pheromone concentrations corresponding to this transition. We fit all the parameters of the mathematical model by using quantitative data on spontaneous polarization in uniform pheromone concentration. Once these parameters have been computed, and without any further fit, our model quantitatively predicts the yeast cell response to pheromone gradient providing an important step toward understanding how cells communicate with each other.


Finding the frame shift: digit loss, developmental variability, and the origin of the avian hand.
The origin of the tridactyl hand of crown birds from the pentadactyl hand of those early theropod dinosaurs lying along the avian stem has become a classic, but at times seemingly intractable, historical problem. The point in question is whether the fingers of crown birds represent digits 1-3 as predicted by generalized trends in the fossil record; or digits 2-4, as evidenced by the topology of the embryonic mesenchymal condensations from which the digits develop. The frame shift hypothesis attempted to resolve this paradox by making these signals congruent by means of a homeotic transformation in digital identity, but recently the paleontological support for this hypothesis was questioned. Here, we reassess the frame shift from a paleontological perspective by addressing what predictions a frame shift makes for skeletal morphology, whether the frame shift remains a viable explanation of the known fossil data, and where on the theropod tree the frame shift most likely occurred. Our results indicate that the frame shift remains viable, and based on the inferred pattern of digit loss, the frame shift likely occurred at a deeper position in theropod phylogeny than previously proposed. A new evolutionary model of the frame shift is described in which the early history of the frame-shifted hand is marked by an extended zone of developmental polymorphism. This model provides a new conceptual framework for the role of developmental variability in communicating broad evolutionary patterns on a taxonomically inclusive phylogenetic tree.


Origins of alphavirus-derived small RNAs in mosquitoes.
The continual transmission in nature of many arthropod-borne viruses depends on the establishment of a persistent, nonpathogenic infection in a mosquito vector. The importance of antiviral immunity directed by small RNAs in the mechanism by which alphaviruses establish a persistent, nonpathogenic infection in the mosquito vector has recently been demonstrated. The origin of the small RNAs central to this RNA silencing response has recently been the subject of debate. Here we briefly summarize what is known about the mechanism of small RNA-directed immunity in invertebrates, and discuss current models for the viral triggers of this response. Finally, we summarize evidence indicating that alphavirus double-stranded replicative intermediates trigger an exogenous-siRNA pathway in mosquitoes resulting in the biogenesis of virus-derived siRNAs.


Socioeconomic and cognitive roots of trait anxiety in young adults.
In 54 participants (41% women) from the Czech arm of the European Longitudinal Study of Pregnancy and Childhood, a national birth cohort with prospectively collected data from their birth until young adulthood, we aimed to study the association between early-life socioeconomic deprivation (ELSD), cognitive ability in adolescence, trait anxiety and resting state functional connectivity of the lateral prefrontal cortex (LPFC) in young adulthood. We found that ELSD was associated with lower cognitive ability in adolescence (at age 13) as well as higher trait anxiety in young adulthood (at age 23/24). Higher cognitive ability in adolescence predicted lower trait anxiety in young adulthood. Resting state functional connectivity between the right LPFC and a cluster of voxels including left precentral gyrus, left postcentral gyrus and superior frontal gyrus mediated the relationship between lower cognitive ability in adolescence and higher trait anxiety in young adulthood. These findings indicate that lower cognitive ability and higher trait anxiety may be both consequences of socioeconomic deprivation in early life. The recruitment of the right LPFC may be the underlying mechanism, through which higher cognitive ability may ameliorate trait anxiety.


Postural orientation: age-related changes in variability and time-to-boundary.
The relation between age-specific postural instability and the detection of stability boundaries was examined. Balance control was investigated under different visual conditions (eyes open/closed) and postural orientations (forward/backward lean) while standing on a force platform. Dependent variables included center of pressure variability and the time-to-contact of the center of pressure with the stability boundaries around the feet (i.e., time-to-boundary). While leaning maximally, older individuals (ages 55-69) showed increased center of pressure variability compared to no lean, while younger subjects (ages 24-38) showed a decrease. These significant differences were found only in anterior-posterior direction. No significant age-specific differences were found between eyes open and eyes closed conditions. Time-to-boundary analysis revealed reduced spatio-temporal stability margins in older individuals in both anterior-posterior and medio-lateral directions. Time-to-boundary variability, however, was not significantly different between the groups in both medio-lateral and anterior-posterior direction. These results show the importance of boundary relevant center of pressure measures in the study of postural control, especially concerning the lateral instability often observed in older adults.


The crossover conformational shift of the GTPase atlastin provides the energy driving ER fusion.
The homotypic fusion of endoplasmic reticulum membranes is catalyzed by the atlastin GTPase. The mechanism involves trans-dimerization between GTPase heads and a favorable crossover conformational shift, catalyzed by GTP hydrolysis, that converts the dimer from a "prefusion" to "postfusion" state. However, whether crossover formation actually energizes fusion remains unclear, as do the sequence of events surrounding it. Here, we made mutations in atlastin to selectively destabilize the crossover conformation and used fluorescence-based kinetic assays to analyze the variants. All variants underwent dimerization and crossover concurrently, and at wild-type rates. However, certain variants were unstable once in the crossover dimer conformation, and crossover dimer stability closely paralleled lipid-mixing activity. Tethering, however, appeared to be unimpaired in all mutant variants. The results suggest that tethering and lipid mixing are catalyzed concurrently by GTP hydrolysis but that the energy requirement for lipid mixing exceeds that for tethering, and the full energy released through crossover formation is necessary for fusion.


The use of alternative health care by a family practice population.
This study examined the characteristics of family practice patients using alternative medicine, the problems that led them to use it, and their satisfaction with its use. A confidential questionnaire was mailed to 250 randomly selected adults enrolled in a large military family practice clinic, with a final response rate of 71 percent. More than 28 percent of patients used some form of alternative medicine. The typical user was 30 to 49 years old, female, white, and well educated. Common methods used were chiropractic (64 percent), massage therapy (36 percent), herbal therapy (32 percent), and acupuncture (16 percent). The most common problems for which patients sought alternative care were back pain (56 percent), other musculoskeletal pain (22 percent), and stress or other psychosocial problems (20 percent). Fewer than one half were satisfied with their alternative health care, although 82 percent reported at least some improvement in their conditions. Most (63 percent) had not told their family physician of their use of alternative health care. A substantial number of family practice patients are using alternative medicine. Although most derive some benefit, most are not satisfied with the results. Reasons for this disparity between satisfaction and effectiveness of alternative medicine deserve further study.


Family-based therapy for dementia caregivers: clinical observations.
Family caregiving for dementia patients is a major social and clinical problem. Family caregivers face major stressful emotional, social and economic burdens, and the negative consequences associated with caregiving are well documented. Given the projected increase in the number of people with dementia, there is a need to identify approaches that will help families manage the challenges of caregiving. Social support from friends and family members has consistently been found to mediate caregiver outcomes, yet many caregivers face problems with isolation and estrangement from family members. In this regard, family-based therapy is a promising intervention for increasing social support for caregivers, and enhancing their quality of life and ability to provide care.This paper will discuss how family-based therapy can be applied as an intervention for family caregivers of dementia patients.The clinical implications of specific interactional patterns will be presented via case examples from an ongoing clinical trial with white American and Cuban American caregivers of dementia patients.The intent is to demonstrate how identification of interactional patterns is a valuable tool for implementing family-based interventions.


Host-recognition of pathogens and commensals in the mammalian intestine.
To peacefully coexist with the microbial inhabitants of the intestine, mammals have evolved elaborate and interconnected regulatory mechanisms to maintain immune homeostasis in the face of potential infection and tissue damage by pathogenic microorganisms. Physical barriers, antimicrobial factors and secretory antibodies act in concert to keep microbes at a distance from the epithelium and initiate repair mechanisms in the event of damage. Commensal bacteria are not ignored but dynamically controlled via many complex overlapping and intertwined mechanisms involving intestinal epithelial cells (IECs) and signals from the microbiota. Polarized IECs play a decisive role in homeostasis by regulating the expression and activity of the pattern-recognition receptors (PRRs), in different compartments of the intestine. The differential signaling and expression of receptors on apical and basal membranes of the epithelium also plays its part in distinguishing commensals from harmful invaders. In steady state conditions macrophages and dendritic cells (DCs) in the lamina propria (LP) are conditioned by environmental factors to induce immune tolerance. The distinction between pathogen and non-pathogen is linked to the ability of pathogens to invade and cause damage to the host cells and tissues. This induces local inflammatory responses and the attraction of capillary leukocytes by chemokines released from colonized and invaded epithelial cells. This bypasses the tolerogenic mechanisms controlling the responses of resident DCs and macrophages leading to pathogen killing and adaptive immune responses. Research on this topic has important implications for the development of novel therapeutic approaches to treat or prevent inflammatory bowel disease (IBD), inflammation-related cancer and other gut-related diseases and disorders.


Clinical and economic impact of Ventricular Assist Device infections: a real-world claims analysis.
VAD therapy has revolutionized the treatment of end-stage heart failure, but infections remain an important complication. The objective of this study was to characterize the clinical and economic impacts of VAD-specific infections. A retrospective analysis of a United States claims database identified members 18 years with a claim for a VAD implant procedure, at least 6 months of pre-implant baseline data, and 12 months of follow-up between June 1, 2016 and December 31, 2019. Cumulative incidence of infection was calculated. Infection and non-infection cohorts were compared regarding mortality, healthcare utilization, and total cost. Regression models were used to identify risk factors associated with infections and mortality. A total of 2,259 patients with a VAD implant were included, with 369 experiencing infection (12-month cumulative incidence 16.1%). Patients with infection were 2.1 times more likely to die (p < 0.001, 95% CI [1.5-2.9]). The mean 12-month total cost per US patient was $354,339 for the non-infection cohort and $397,546 for the infection cohort, a difference of $43,207 (p < 0.0001). VAD infections were associated with higher mortality, more healthcare utilization, and higher total cost. Strategies to minimize VAD-specific infections could lead to improved clinical and economic outcomes.


The influence of HIV-related stigma on PrEP disclosure and adherence among adolescent girls and young women in HPTN 082: a qualitative study.
Stigma and disclosure concerns have been key barriers to oral pre-exposure prophylaxis (PrEP) adherence for African adolescent girls and young women (AGYW) in efficacy trials. We aimed to understand the impact of these factors among African AGYW in an open-label PrEP study. HPTN 082 was an open-label PrEP study among AGYW (ages 16 to 24) in Harare, Zimbabwe, and Cape Town and Johannesburg, South Africa from 2016 to 2018. Women starting PrEP were randomized to standard adherence support (counselling, two-way SMS, monthly adherence clubs) or standard support plus drug-level feedback. Serial in-depth interviews were conducted among 67 AGYW after 13-week and 26-week study visits to explore experiences of stigma, disclosure and PrEP adherence. We analysed data by coding transcripts and memo-writing and diagramming to summarize themes. AGYW described stigma related to sexual activity (e.g. "people say I'm a prostitute") and being perceived to be living with HIV because of taking antiretrovirals (e.g. "my husband's friends say I'm HIV infected"). Participants who anticipated stigma were reluctant to disclose PrEP use and reported adherence challenges. Disclosure also resulted in stigmatizing experiences. Across all sites, negative descriptions of stigma and disclosure challenges were more common in the first interview. In the second interview, participants often described disclosure as an "empowering" way to combat community-level PrEP stigma; many said that they proactively discussed PrEP in their communities (e.g. became a "community PrEP ambassador"), which improved their ability to take PrEP and encourage others to use PrEP. These empowering disclosure experiences were facilitated by ongoing HPTN 082 study activities (e.g. counselling sessions, adherence clubs) in which they could discuss PrEP-related stigma, disclosure and PrEP adherence issues. Stigma and disclosure challenges were initial concerns for African AGYW newly initiating PrEP but many were empowered to disclose PrEP use over their first six months of PrEP use, which helped them cope with stigma and feel more able to take PrEP regularly. PrEP programmes can foster disclosure through community and clinic-based discussion, adherence clubs and activities normalizing sexual behaviour and PrEP use, which can reduce stigma and improve PrEP adherence and thus effectiveness.


Tuberculosis risk factors and mortality for HIV-infected persons receiving antiretroviral therapy in South Africa.
To determine important risk factors for and impact of tuberculosis on survival in HIV-infected patients starting antiretroviral therapy (ART) in South Africa. Prospective trial of 1771 HIV-infected patients with either CD4 cell count less than 200 cells/microl or a prior AIDS-defining illness, enrolled in randomized trial of four antiretroviral regimens. Data collected from patient records. A history of tuberculosis at study entry was reported by 27% of patients and correlated with poor baseline health status. A history of tuberculosis at baseline was associated with subsequent tuberculosis and death during ART, but was not itself an independent risk factor for poor outcome. Tuberculosis was diagnosed during ART in 14% of patients and was more frequent during the first 3 months. Tuberculosis during therapy was independently associated with increased hazard of other AIDS-defining events and death, regardless of when during ART tuberculosis occurred. ART that consistently suppressed circulating viremia reduced but did not eliminate tuberculosis risk. In HIV-infected patients who started ART at low CD4 cell counts, tuberculosis at baseline was a predictor of death, but was not independent of other factors indicating poor baseline health status. Tuberculosis during follow-up was, in contrast, an independent predictor of death even after adjustments for baseline risk factors, including CD4 cell count and viral load. Virologic failure during ART was associated with a 55% increase in risk of tuberculosis. Thus, tuberculosis is a major marker for poor outcome both at baseline and during ART and is not completely eliminated by fully suppressive ART.


Surgery for fistula-in-ano in a specialist colorectal unit: a critical appraisal.
Several techniques have been described for the management of fistula-in-ano, but all carry their own risks of recurrence and incontinence. We conducted a prospective study to assess type of presentation, treatment strategy and outcome over a 5-year period. Between 1st January 2005 and 31st March 2011,247 patients presenting with anal fistulas were treated at the University Hospital Tor Vergata and were included in the present prospective study. Mean age was 47 years (range 16-76 years); minimum follow-up period was 6 months (mean 40, range 6-74 months).Patients were treated using 4 operative approaches: fistulotomy, fistulectomy, seton placement and rectal advancement flap. Data analyzed included: age, gender, type of fistula, operative intervention, healing rate, postoperative complications, reinterventions and recurrence. Etiologies of fistulas were cryptoglandular (n = 218), Crohn's disease (n = 26) and Ulcerative Colitis (n = 3). Fistulae were classified as simple -intersphincteric 57 (23%), low transphincteric 28 (11%) and complex -high transphicteric 122 (49%), suprasphincteric 2 (0.8%), extrasphinteric 2 (0.8%), recto-vaginal 7 (2.8%) Crohn 26 (10%) and UC 3 (1.2%).The most common surgical procedure was the placement of seton (62%), usually applied in case of complex fistulae and Crohn's patients.Eighty-five patients (34%) underwent fistulotomy, mainly for intersphincteric and mid/low transphincteric tracts. Crohn's patients were submitted to placement of one or more loose setons.The main treatment successfully eradicated the primary fistula tract in 151/247 patients (61%). Three cases of major incontinence (1.3%) were detected during the follow-up period; Furthermore, three patients complained minor incontinence that was successfully treated by biofeedback and permacol injection into the internal anal sphincter. This prospective audit demonstrates an high proportion of complex anal fistulae treated by seton placement that was the most common surgical technique adopted to treat our patients as a first line. Nevertheless, a good outcome was achieved in the majority of patients with a limited rate of faecal incontinence (6/247 = 2.4%). New technologies provide promising alternatives to traditional methods of management particularly in case of complex fistulas. There is, however, a real need for high-quality randomized control trials to evaluate the different surgical and non surgical treatment options.


[Malignant adenoma of the cervix: a diagnostic and therapeutic dilemma].
"Adenoma malignum" of the cervix is an extremely rare variant of cervical adenocarcinoma. Its diagnosis by means of light microscopy is rendered very difficult by its strong similarity to benign endocervical hyperplasia. During the last few years immunohistochemistry has proved to be of great help in differential diagnosis. Treatment of "adenoma malignum" also poses great problems. Most cases are diagnosed in advanced stages, although only early diagnosis offers the possibility of effective treatment. Radiotherapy has failed in all known cases in an advanced stage of the disease. Chemotherapy is also not promising as a therapeutical option, because the tumour expresses high levels of the multidrug-resistance marker P-glycoprotein.


Cardioprotective effect of virgin coconut oil in heated palm oil diet-induced hypertensive rats.
Virgin coconut oil (VCO) contains high antioxidant activity which may have protective effects on the heart in hypertensive rats. The study investigated the effects of VCO on blood pressure and cardiac tissue by measuring angiotensin-converting enzyme (ACE) activity and its histomorphometry in rats fed with a heated palm oil (HPO) diet. Thirty-two male Sprague-Dawley rats were randomly divided into four groups: (i) control, (ii) orally given VCO (1.42 ml/kg), (iii) fed with a HPO (15%) diet, and (iv) fed with a HPO diet and supplemented with VCO (1.42 ml/kg, po) (HPO+VCO) for 16 weeks. Blood pressure was measured monthly. After 16 weeks, rat hearts were dissected for lipid peroxidation (TBARS) and ACE activity measurement and histomorphometric study. Systolic blood pressure was significantly increased in the HPO group compared with the control starting at week eight (112.91 ± 1.32 versus 98.08 ± 3.61 mmHg, p < 0.05) which was prevented by VCO supplementation (91.73 ± 3.42 mmHg). The consumption of HPO increased TBARS and ACE activity in heart, which were inhibited by VCO supplementation. The increases in the myofiber width and area as well as nuclear size reduction in the HPO group were significantly prevented by VCO supplementation. These results suggested that VCO supplementation possesses a cardioprotective effect by preventing the increase in blood pressure via an antioxidant mechanism and remodeling in rats fed repeatedly with a HPO diet.


[Hepatic cell transformation changes in experimental opisthorchiasis].
Changes in the liver where Opisthorchis felines (Rivolta, 1884) parasitizes were traced by light and transmission electron microscopies on experimental models (gold hamsters) of acute and chronic opisthorchiasis. Regenerative processes, such as hepatic epithelial mitotic indices, binucleated, hypertrophic hepatocytes, were determined; the composition of infiltrates was identified; fibroplastic changes and hepatocytic apoptotic index were traced. The progression/regression (P/R) ratio was used to detect atrophy of a hepatic parenchymatous component (hepatocytes) in different periods (from 3 to 120 days) of invasion. In opisthorchiasis, proliferation of committed hepatocytes was first found on day 10 of invasion. Oval (committed) cells are differentiated mainly to cholangiocytes, followed by the formation of ductular structures; the committed cells of the liver may be simultaneously a source of programmed hepatocellular differentiation; however, the newly formed hepatocytes are solitary amongst them in opisthorchiasis.


[The parenchymal cellular composition and extracellular matrix in ontogenesis of Triaenophorus nodulus (Cestoda)].
The cellular composition of parenchyma was studied in the ontogenesis of Triaenophorus nodulosus and it was shown that the specialized parenchymal cells was absent. The extracellular matrix fibrils were synthesized successively by the tegumental cytons and muscle cells. The coracidium basal matrix consisted of electron-light and electron-dense layers, and the reticular layer appeared at the procercoid stage. Reserve nutrients were accumulated by the musculocutaneous sac elements at the procercoid stage and, later, by the glandular and tegumental cells. It has been proposed that the parenchyma as an independent histological unit is absent, while the parenchymal organization is based on all specialized cellular elements that do not lose their typical features.


A pilot study of cognitive therapy in bipolar disorders.
The efficacy and effectiveness of cognitive therapy (CT) is well established for unipolar disorders, but little is known about its utility in bipolar disorders. This study aimed to explore the feasibility and efficacy of using CT as an adjunct to usual psychiatric treatment in this patient population. Subjects referred by general adult psychiatrists were assessed by and independent rater and then randomly allocated to immediate CT (N = 21) or 6-month waiting-list control, which was then followed by CT (N = 21). Observer and self-ratings of symptoms and functioning were undertaken immediately prior to CT, after a 6-month course of CT and a further 6-months later. Data on relapse and hospitalization rates in the 18 months before and after commencing CT were also collected. At 6-month follow-up, subjects allocated to CT showed statistically significantly greater improvements in symptoms and functioning as measured on the Beck Depression Inventory, the Internal State Scale, and the Global Assessment of Functioning than those in the waiting-list control group. In the 29 patients who eventually received CT, relapse rates in the 1 8 months after commencing CT showed a 60 % reduction in comparison with the 18 months prior to commencing CT. Seventy per cent of subjects who commenced therapy viewed CT as highly acceptable. Although the results of this study are encouraging, the use of CT in subjects with bipolar disorders is more complex than in unipolar disorders and requires a high level of therapist expertise. The therapy may prove to be particularly useful in the treatment of bipolar depression.


Microstructure-dependent CO2-responsive microemulsions for deep-cleaning of oil-contaminated soils.
CO2-responsive microemulsion (ME) is considered a promising candidate for deep-cleaning and oil recovery from oil-contaminated soils. Understanding the responsive nature of different microstructures (i.e., oil-in-water (O/W), bicontinuous (B.C.) and water-in-oil (W/O)) is essential for unlocking the potential and mechanisms of CO2-responsive emulsions in complex multiphase systems and providing comprehensive guidance for remediation of oil-contaminated soils. Herein, the responsiveness of microstructures of ME to CO2 trigger was investigated using experimental designs and coarse-grained molecular dynamic simulations. MEs were formed for the first time by a weakly associated pseudo-Gemini surfactant of indigenous organic acids (naphthenic acids, NAs are a class of natural surface-active molecules in crude oil) and tetraethylenepentamine (TEPA) through fine tuning of co-solvent of dodecyl benzene sulfonic acid (DBSA) and butanol. The O/W ME exhibited an optimal CO2-responsive character due to easier proton migration in the continuous aqueous phase and more pronounced dependence of configuration on deprotonated NA ions. Conversely, the ME with W/O microstructure exhibited a weak to none responsive characteristic, most likely attributed to its high viscosity and strong oil-NA interactions. The O/W ME also showed superior cleaning efficiency and oil recovery from oil-contaminated soils. The results from this study provide insights for the design of CO2-responsive MEs with desired performance and guidance for choosing the favorable operating conditions in various industrial applications, such as oily solid waste treatment, enhanced oil recovery (EOR), and pipeline transportation. The insights from this work allow more efficient and tailored design of switchable MEs for manufacturing advanced responsive materials in various industrial sectors and formulation of household products.


Coping with early stage breast cancer: examining the influence of personality traits and interpersonal closeness.
The study examines the influence of personality traits and close relationships on the coping style of women with breast cancer. A sample of 72 Italian patients receiving treatment for early stage breast cancer was recruited. Participants completed questionnaires measuring personality traits (Interpersonal Adaptation Questionnaire), interpersonal closeness (Inclusion of the Other in the Self Scale), and adjustment to cancer (Mini-Mental Adjustment to Cancer Scale). We hypothesized that diverse personality traits and degrees of closeness contribute to determine the coping styles shown by participants. Multiple regression analyses were conducted for each of the five coping styles (Helplessness/Hopelessness, Anxious Preoccupation, Avoidance, Fatalism, and Fighting Spirit) using personality traits and interpersonal closeness variables (Strength of Support Relations, and Number of Support Relations) as predictors. Women who rated high on assertiveness and social anxiety were more likely to utilize active coping strategies (Fighting Spirit). Perceived strength of relationships was predictive of using an active coping style while the number of supportive relationships did not correlate with any of the coping styles. Implications for assessment of breast cancer patients at risk for negative adaptation to the illness and the development of psychosocial interventions are discussed.


Distribution of parkin in the adult rat brain.
A mutation in the parkin gene has been identified as the cause for an autosomal recessively inherited form of Parkinson's disease (PD). The authors have recently isolated the mRNA coding for the rat homolog of parkin and showed its widespread expression in the central nervous system (CNS) by in situ hybridization. In the present study, we investigated the distribution of parkin in the rat CNS with a polyclonal antibody that reacts with a approximately 52-kDa protein, mainly localized in the cytoplasm and corresponding to the predicted molecular mass of parkin. Immunohistochemistry on adult rat brain sections showed a widespread distribution of parkin. This included labeling of cell bodies, nuclei as well as processes in the hippocampus, cerebral cortex, cerebellum, and several nuclei in the brainstem. The regional expression of parkin-immunoreactivity (IR) correlated well with the parkin-mRNA levels assessed by real-time quantitative reverse transcription-polymerase chain reaction (RT-PCR). This study provides the detailed analysis of the regional and cellular distribution of parkin in the rat brain and may be useful in elucidating its pathophysiological role.


Structural basis of the XPB helicase-Bax1 nuclease complex interacting with the repair bubble DNA.
Nucleotide excision repair (NER) removes various DNA lesions caused by UV light and chemical carcinogens. The DNA helicase XPB plays a key role in DNA opening and coordinating damage incision by nucleases during NER, but the underlying mechanisms remain unclear. Here, we report crystal structures of XPB from Sulfurisphaera tokodaii (St) bound to the nuclease Bax1 and their complex with a bubble DNA having one arm unwound in the crystal. StXPB and Bax1 together spirally encircle 10 base pairs of duplex DNA at the double-/single-stranded (ds-ss) junction. Furthermore, StXPB has its ThM motif intruding between the two DNA strands and gripping the 3'-overhang while Bax1 interacts with the 5'-overhang. This ternary complex likely reflects the state of repair bubble extension by the XPB and nuclease machine. ATP binding and hydrolysis by StXPB could lead to a spiral translocation along dsDNA and DNA strand separation by the ThM motif, revealing an unconventional DNA unwinding mechanism. Interestingly, the DNA is kept away from the nuclease domain of Bax1, potentially preventing DNA incision by Bax1 during repair bubble extension.


Antioxidants inhibit neuronal toxicity in Parkinson's disease-linked LRRK2.
Mutations in leucine-rich repeat kinase-2 are the most common cause of familial Parkinson's disease. The prevalent G2019S mutation increase oxidative, kinase and toxic activity and inhibit endogenous peroxidases. We initially screened a library of 84 antioxidants and identified seven phenolic compounds that inhibited kinase activity on leucine-rich repeat kinase-2 substrates. The representative antioxidants (piceatannol, thymoquinone, and esculetin) with strong kinase inhibitor activity, reduced loss in dopaminergic neurons, oxidative dysfunction, and locomotor defects in G2019S-expressing neuronal and Drosophila models compared to weak inhibitors. We provide proof of principle that natural antioxidants with dual antioxidant and kinase inhibitor properties could be useful for leucine-rich repeat kinase-2-linked Parkinson's disease.


Review article: defining remission in ulcerative colitis.
There is no international agreement on scoring systems used to measure disease activity in ulcerative colitis, nor is there a validated definition for disease remission. To review the principles and components for defining remission in ulcerative colitis and propose a definition that will help improve patient outcomes. A review of current standards of remission from the perspective of clinical trials, guidelines, clinical practice and patients was conducted by the authors. Selected literature focused on the components of a definition of remission, the utility of a definition and treatment strategies, based on current definitions. Different definitions of remission affect the assessment of outcome and make it difficult to compare trials. In the clinic, endoscopy is rarely used to confirm remission, because mucosal healing has only recently begun to be related to the duration of subsequent remission in a way that will affect clinical practice. Histopathology may be the ultimate arbiter of mucosal healing. There is no agreement on the definition of remission in current guidelines. Patient-defined remission may predict endoscopic remission, but has yet to be shown to predict duration of remission. A standard based on clinical symptoms and endoscopy is proposed. Histopathology is a third dimension of remission that may have prognostic value. The definition of remission should help predict long-term outcome. The expectations of patients and their physicians need to be raised, as the goal of treatment of active ulcerative colitis should be to induce remission.


Regulation of human brown adipose tissue by adenosine and A2A receptors - studies with [15O]H2O and [11C]TMSX PET/CT.
Brown adipose tissue (BAT) has emerged as a potential target to combat obesity and diabetes, but novel strategies to activate BAT are needed. Adenosine and A2A receptor (A2AR) agonism activate BAT in rodents, and endogenous adenosine is released locally in BAT as a by-product of noradrenaline, but physiological data from humans is lacking. The purpose of this pilot study was to investigate the effects of exogenous adenosine on human BAT perfusion, and to determine the density of A2ARs in human BAT in vivo for the first time, using PET/CT imaging. Healthy, lean men (n = 10) participated in PET/CT imaging with two radioligands. Perfusion of BAT, white adipose tissue (WAT) and muscle was quantified with [15O]H2O at baseline, during cold exposure and during intravenous administration of adenosine. A2AR density of the tissues was quantified with [11C]TMSX at baseline and during cold exposure. Adenosine increased the perfusion of BAT even more than cold exposure (baseline 8.3 ± 4.5, cold 19.6 ± 9.3, adenosine 28.6 ± 7.9 ml/100 g/min, p < 0.01). Distribution volume of [11C]TMSX in BAT was significantly lower during cold exposure compared to baseline. In cold, low [11C]TMSX binding coincided with high concentrations of noradrenaline. Adenosine administration caused a maximal perfusion effect in human supraclavicular BAT, indicating increased oxidative metabolism. Cold exposure increased noradrenaline concentrations and decreased the density of A2AR available for radioligand binding in BAT, suggesting augmented release of endogenous adenosine. Our results show that adenosine and A2AR are relevant for activation of human BAT, and A2AR provides a future target for enhancing BAT metabolism.


Reversible induction of translational isoforms of p53 in glucose deprivation.
Tumor suppressor protein p53 is a master transcription regulator, indispensable for controlling several cellular pathways. Earlier work in our laboratory led to the identification of dual internal ribosome entry site (IRES) structure of p53 mRNA that regulates translation of full-length p53 and Δ40p53. IRES-mediated translation of both isoforms is enhanced under different stress conditions that induce DNA damage, ionizing radiation and endoplasmic reticulum stress, oncogene-induced senescence and cancer. In this study, we addressed nutrient-mediated translational regulation of p53 mRNA using glucose depletion. In cell lines, this nutrient-depletion stress relatively induced p53 IRES activities from bicistronic reporter constructs with concomitant increase in levels of p53 isoforms. Surprisingly, we found scaffold/matrix attachment region-binding protein 1 (SMAR1), a predominantly nuclear protein is abundant in the cytoplasm under glucose deprivation. Importantly under these conditions polypyrimidine-tract-binding protein, an established p53 ITAF did not show nuclear-cytoplasmic relocalization highlighting the novelty of SMAR1-mediated control in stress. In vivo studies in mice revealed starvation-induced increase in SMAR1, p53 and Δ40p53 levels that was reversible on dietary replenishment. SMAR1 associated with p53 IRES sequences ex vivo, with an increase in interaction on glucose starvation. RNAi-mediated-transient SMAR1 knockdown decreased p53 IRES activities in normal conditions and under glucose deprivation, this being reflected in changes in mRNAs in the p53 and Δ40p53 target genes involved in cell-cycle arrest, metabolism and apoptosis such as p21, TIGAR and Bax. This study provides a new physiological insight into the regulation of this critical tumor suppressor in nutrient starvation, also suggesting important functions of the p53 isoforms in these conditions as evident from the downstream transcriptional target activation.


Short-Term Propofol Infusion and Associated Effects on Serum Lactate in Pediatric Patients.
This study aimed to determine if short-duration procedural sedation in children with propofol is related to an adverse metabolic stress response measured by serum lactate. Propofol infusion syndrome is associated with high-dose and long-duration infusion. It has not been studied with short-duration, outpatient propofol administration. This was a prospective, longitudinal study that included 50 patients between the ages of 2 and 18 years who were undergoing outpatient procedural sedation with propofol at a pediatric sedation unit. Patients received bolus or bolus and continuous infusion doses of propofol. Serum lactate values were obtained before and after the patients' sedated procedures. The average length of procedure was 40.3 minutes, and the mean dose of propofol per patient was 8.2 mg/kg. The highest measured lactate value was 1.8 mmol/L. The average (SD) preprocedure and postprocedure lactate values were 1.0 (0.3) and 0.7 (0.2) mmol/L, respectively, resulting in an overall significant decrease in lactate of -0.3 (0.3) mmol/L (P < 0.001). There was a significant (P = 0.01) positive relationship between age and postprocedure lactate value, when controlling for the length of the procedure in a multivariable regression. No significant association was found between propofol dosage and length of the procedure with lactate values. In this study, we did not find a significant increase in metabolic stress, measured by serum lactate. Using propofol for short-duration procedural sedation may not carry similar risks for propofol infusion syndrome to those for long-duration/high-dose infusion therapy.


Challenges in Radiofrequency Pasteurization of Shell Eggs: Coagulation Rings.
A total of 50 different configurations of simple radiofrequency (RF) heating at 27.12 MHz of a shell egg were simulated using a finite element model with the purpose of pasteurizing the egg. Temperature-dependent thermal and dielectric properties of the yolk, albumen, and shell were measured, fitted, and introduced into the model. A regression equation that relates the top electrode voltage to the gap between the electrodes and vertical position of the egg was developed. Simulation and experimental results had good agreement in terms of temperature deviation (root mean squared error ranged from 0.35 °C to 0.48 °C) and both results demonstrated the development of a "coagulation ring" around the air cell. The focused heating near the air cell of the egg prevented pasteurization of the egg due to its impact on quality (coagulation). Analysis of the electric field patterns offered a perspective on how nonuniform RF heating could occur in heterogeneous food products. The results can be used to guide development of RF heating for heterogeneous food products and further development of RF pasteurization of eggs.


Anaphylaxis and anaphylactoid reactions associated with the insertion of peripherally inserted central catheters: A multiyear comparative retrospective cohort study.
Peripherally inserted central catheters (PICCs) are a mainstay of nonpermanent vascular access devices. In this study, we assessed patients displaying anaphylaxis or anaphylactoid reactions to the PowerPICC SOLO and Groshong PICC (Bard Access Systems) using the Sherlock tip locating system (TLS). Patients from 2 tertiary-care hospitals were systematically monitored over 4 years for adverse events following the insertion of a PICC using the Sherlock TLS. Insertion data were also collected using the BioFlo PICC (Angiodynamics)from a third hospital site and from The Ottawa Hospital over 4 years as an additional comparator. Three definitions of anaphylaxis and anaphylactoid reactions were utilized, and the Cohen κ was used to assess interrater agreement. Analysis of reactions among the patient cohorts was performed using the χ2 test with Yates correction or the Fisher exact test as appropriate. Among 8,257 insertions using the TLS PICCs, 37 potential reactions (0.45%) were recorded. Using specific definitions for anaphylaxis or anaphylactoid reactions, 54.1%-91.9% met criteria. Comparator populations using data from Calgary (n = 491) and Ottawa (n = 7,889) using the BioFlo PICC insertion found no reactions. Anaphylactic or anaphylactoid reactions were significantly associated with the PowerPICC SOLO and Groshong PICC with the TLS compared to the BioFlo PICC (P < .0001) across all definitions. The largest subset of patients experiencing adverse reactions had cystic fibrosis (CF) (n = 4, 10.8%). Our study results demonstrate significant adverse events associated with the PowerPICC SOLO and Groshong PICC using the Sherlock TLS inserted across a range of patient populations. The incidence rate of anaphylaxis or anaphylactoid reactions in the CF population at our center is significantly higher than in non-CF patients (P < .001).


What do we need to consider when planning, implementing and researching the use of alternatives to face-to-face consultations in primary healthcare?
Communications technologies are variably utilised in healthcare. Policymakers globally have espoused the potential benefits of alternatives to face-to-face consultations, but research is in its infancy. The aim of this essay is to provide thinking tools for policymakers, practitioners and researchers who are involved in planning, implementing and evaluating alternative forms of consultation in primary care. We draw on preparations for a focussed ethnographic study being conducted in eight general practice settings in the UK, knowledge of the literature, qualitative social science and Cochrane reviews. In this essay we consider different types of patients, and also reflect on how the work, practice and professional identities of different members of staff in primary care might be affected. Elements of practice are inevitably lost when consultations are no longer face-to-face, and we know little about the impact on core aspects of the primary care relationship. Resistance to change is normal and concerns about the introduction of alternative methods of consultation are often expressed using proxy reasons; for example, concerns about patient safety. Any planning or research in the field of new technologies should be attuned to the potential for unintended consequences. Implementation of alternatives to the face-to-face consultation is more likely to succeed if approached as co-designed initiatives that start with the least controversial and most promising changes for the practice. Researchers and evaluators should explore actual experiences of the different consultation types amongst patients and the primary care team rather than hypothetical perspectives.


Semicarbazide-sensitive amine oxidases in heart and bovine serum.
In guinea pig dorsal skin the semicarbazide-sensitive amine oxidase (SSAO) is localised in fibroblasts. Fibroblasts in culture lose the ability to express this enzymatic activity with doublings, thus suggesting that the SSAO expression needs some factors which are not present in the 10% bovine serum culture medium. Fresh bovine serum of adult animals contains two SSAO activities, one with high affinity for benzylamine and one with lower affinity. The enzyme with lower affinity for benzylamine was identified as spermine oxidase, the oxidation of [14C]-benzylamine was inhibited by semicarbazide, alpha-aminoguanidine and B24, a specific inhibitor of benzylamine oxidase and spermine oxidase, both SSAO enzymes. The enzymatic activity of bovine serum was partially purified, the kinetic properties and sensitivity to inhibitors studied. A mathematical procedure for the analysis of the kinetics resulting from the activity of two enzymes acting on the same substrate seems to give better results than the methods previously described.


PET, image-guided HDAC inhibition of pediatric diffuse midline glioma improves survival in murine models.
Efforts at altering the dismal prognosis of pediatric midline gliomas focus on direct delivery strategies like convection-enhanced delivery (CED), where a cannula is implanted into tumor. Successful CED treatments require confirmation of tumor coverage, dosimetry, and longitudinal in vivo pharmacokinetic monitoring. These properties would be best determined clinically with image-guided dosimetry using theranostic agents. In this study, we combine CED with novel, molecular-grade positron emission tomography (PET) imaging and show how PETobinostat, a novel PET-imageable HDAC inhibitor, is effective against DIPG models. PET data reveal that CED has significant mouse-to-mouse variability; imaging is used to modulate CED infusions to maximize tumor saturation. The use of PET-guided CED results in survival prolongation in mouse models; imaging shows the need of CED to achieve high brain concentrations. This work demonstrates how personalized image-guided drug delivery may be useful in potentiating CED-based treatment algorithms and supports a foundation for clinical translation of PETobinostat.


c-Kit-dependent growth of uveal melanoma cells: a potential therapeutic target?
This study was conducted to investigate the expression and functional impact of the proto-oncogene c-kit in uveal melanoma. Based on immunohistochemical (IHC) study of paraffin-embedded specimens from 134 uveal melanomas and Western blot analysis on eight fresh-frozen samples the expression of c-kit in uveal melanoma was studied. Furthermore, the phosphorylation of c-kit and the impact of the tyrosine kinase inhibitor STI571 was examined in the three uveal melanoma cell lines OCM-1, OCM-3, and 92-1. Eighty-four of 134 paraffin-embedded samples and six of eight fresh-frozen samples expressed c-kit. c-Kit was strongly expressed and tyrosine phosphorylated in cultured uveal melanoma cells compared with cutaneous melanoma cells. Moreover, in contrast to cutaneous melanoma cell lines c-kit maintained a high phosphorylation level in serum-depleted uveal melanoma cells. No activation-related mutations in exon 11 of the KIT gene were found. On the contrary, expression of the stem cell growth factor (c-kit ligand) was detected in all three uveal melanoma cell lines, suggesting the presence of autocrine (paracrine) stimulation pathways. Treatment of uveal melanoma cell lines with STI571, which blocks c-kit autophosphorylation, resulted in cell death. The IC(50) of the inhibitory effects on c-kit phosphorylation and cell proliferation was of equal size and less than 2.5 microM. The results confirm that c-kit is vastly expressed in uveal melanoma, suggest that the c-kit molecular pathway may be important in uveal melanoma growth, and point to its use as a target for therapy with STI571.


[Distribution, abundance and population parameters of Diapterus rhombeus (Pisces: Gerreidae) in Laguna de Términos, Campeche, Mexico].
Diapterus rhombeus dominates the nektonic community of Terminos Lagoon and is associated to zones with submerged vegetation, temperatures between 27 and 33 degrees C and salinity between 25 and 36.5 UPS. A total of 3,024 individuals were collected with a trawl net in 23 sites between September 1997 and September 1998. The length-weight model parameters are a = 0.0098; b = 3.155 with a 96% correlation. The growth parameters are L infinity = 20.4 cm, k = 0.74 and t(o) = -4.207 and the total mortality coefficient is 3.526. Fish length is 2.5-18.0 cm with a mode of 7.5 cm. Recruitment is continuous with a peak from May to August. The species uses several habitats of the lagoon mainly for feeding and growth, reproduction probably occurs in the adjacent continental shelf. Movements between continental shelf and the estuarine system represent a continuous energy flow.


ACTH-related peptides, kindling and seizure disorders.
Very small amounts of pituitary hormones and their peptide fragments can profoundly affect learning, memory and other behaviors in both rodents and humans. In addition, several potent pituitary hormone analogs have been developed (e.g. ORG-2766) which retain the behavioral but not the endocrine properties of the parent hormone. The abilities of these peptides to influence nervous system functions suggested that they also may be capable of modifying seizure activity. This possibility is supported by the fact that treatment with adrenocorticotropic hormone (ACTH) represents a major effective therapy for at least one clinical convulsive disorder, infantile spasm. This contention also is reinforced by our findings that ORG-2766 markedly reduces both the behavioral severity and the spread of seizure potentials in an animal model of epilepsy, the kindled rat. By contrast, arginine vasopressin and its non-endocrine desglycyl fragment (DGAVP) facilitates the seizure process in this animal model. Our research also suggests that the behavioral and physiological effects of certain anterior and posterior pituitary hormone fragments depend, in part, on their abilities to modulate permeability mechanisms in brain vasculature. In addition, and especially significant from the standpoint of etiology, is the observation that the kindling process itself appears to alter cerebrovascular permeability. In the kindled rat, "permanent" decreases in permeability (60-75%) are found selectively in the hypothalamic and hippocampal regions, weeks after the last seizure. Several lines of evidence indicate that disruption of normal cerebrovascular function occurs following epileptiform seizures. We propose that an immediate increase in cerebral blood flow and cerebrovascular permeability occurs following a single, acute seizure, and that repeated chronic seizures lead to damage to the cerebrovascular system. Sustained damage would be expected to contribute to the development and maintenance of a chronic seizure focus. Such observations suggest a link between important cerebrovascular disturbances associated with seizures and the existing known and proposed electrophysiological, metabolic and neuropathological substrates of epilepsy. They also point to new strategies for the treatment of seizure disorders by focusing on ways to reduce or prevent cerebrovascular damage.


Predicting cardiotoxicity propensity of the novel iodinated contrast medium GE-145: Ventricular fibrillation during left coronary arteriography in pigs.
Severe side effects caused by iodinated radiographic contrast media (CM) are rare, but can occur in high risk patients and during percutaneous coronary intervention. To minimize this risk a new nonionic CM with low inherent osmolality has been designed, giving room for a relatively high concentration of favorable electrolytes in the isotonic formulation. To test a new radiographic CM (GE-145) in a pig model of cardiotoxicity by comparing its ventricular fibrillation (VF) propensity and hemodynamic effects to that of iodixanol. Test agents were injected into the left anterior descending coronary artery (LAD) of pigs through an inflated balloon catheter (injection volume 25 ml, injection rate 0.4 ml/s, maximum injection time 62.5 s). Series 1: GE-145 (338 mg I/ml) + 45 mM NaCl and iodixanol (321 mg I/ml) + 19 mM NaCl were injected in five pigs. Series 2: GE-145 (320 mg I/ml) + 45 mM NaCl + 0.1, 0.3, or 0.7 mM CaCl₂ and iodixanol (320 mg I/ml) + 19 mM NaCl + 0.3 mM CaC₂ (Visipaque) were injected in six pigs. Iodixanol + NaCl caused VF in 6 of 13 injections (46%) after 60.3±7.5 s (mean ± SD). GE-145 + NaCl did not cause any VF in 13 injections (0%) (P<0.05). Iodixanol + 19 mM NaCl + 0.3 mM CaCl₂ caused VF in 9 of 9 injections (100%) after 61±4 s. GE-145 + 45 mM NaCl + 0.1, 0.3, or 0.7 mM CaCl₂ did not cause any VF during or after 9 injections of each agent (0%) (P<0.05). The least hemodynamic effects were seen with GE-145 + 45 mM NaCl + 0.7 mM CaCl₂. In this model of direct administration of CM into the LAD of anesthetized pigs, the tested GE-145 formulations had a significantly lower propensity to induce VF than iodixanol with electrolytes. Favorable hemodynamic properties of GE-145 can be achieved by optimizing concentrations of sodium and calcium.


Long-term outcomes of endoscopic submucosal dissection for colorectal epithelial neoplasms.
Endoscopic submucosal dissection (ESD) provides a high en bloc resection rate with less invasiveness than surgical resection for large or scarring gastrointestinal neoplasms. However, detailed outcomes in colorectal ESD are still lacking. The aim of our study was to elucidate short- and long-term outcomes of colorectal ESD. 310 consecutive colorectal epithelial neoplasms (146 adenomas, 164 carcinomas), in 290 patients, which fulfilled our indication criteria and were treated with ESD between July 2000 and December 2008 were studied. ESD was done by three skilled endoscopists. As short-term outcomes, rates of en bloc resection, en bloc plus R0 resection, and major complications were analyzed. As long-term outcomes, disease-free and overall survival were assessed in 224 patients. Rates of en bloc resection and en bloc plus R0 resection were 90.3 % and 74.5 %, respectively. Eight patients underwent additional colectomy due to histopathologically proven possible node-positive cancer. Intraoperative perforations occurred with 14 lesions (4.5 %), which were treated successfully only by endoscopic clipping. Emergent surgery was needed for one case of postoperative perforation. Blood transfusion due to intraoperative massive bleeding was required in 1 case (0.3 %). Postoperative bleeding occurred with four lesions (1.3 %), and was endoscopically managed without blood transfusion. Local recurrence was detected in 4 lesions (4/202 patients, 2.0 %); resection had been piecemeal in all 4. During a median follow-up of 38.7 months (range 12.8 - 104.2), the 3- and 5-year overall/disease-specific survivals were 97.1/100 % and 95.3/100 %, respectively. Colorectal ESD showed favorable long-term outcomes. It may largely replace colectomy for node-negative colorectal epithelial neoplasia.


Primary thyroid lymphoma: A rare disease.
Primary thyroid lymphomas are rare neoplasms comprising of 1-5% of thyroid malignancies. These are predominantly B-cell in origin. Here, we report a case of 60 years lady, a known case of lymphocytic thyroiditis, diagnosed as thyroid lymphoma (diffuse large B-cell) on fine needle aspiration and confirmed histopathogically and immunohistochemically. She presented with a sudden increase in thyroid swelling. Fine needle aspiration performed showed highly cellular smears comprising predominantly of the monomorphic population of medium to large sized lymphoid cells with high nuclear/cytoplasmic ratio and scant cytoplasm. A possibility of thyroid lymphoma possibly diffuse large B-cell lymphoma was suggested which was later confirmed on biopsy. Fine needle aspiration provides an easy mode for diagnosing large cell lymphoma like diffuse large B-cell. Hence, an early diagnosis is possible for a timely intervention. Also, cases of lymphocytic thyroiditis should be regularly followed for the development of lymphoma.


The relationship between wellness and training and match load in professional male soccer players.
The aims of this study were to: (i) analyse the within-microcycle variations in professional soccer players; (ii) analyse the relationships between wellness and training and match load demands; (iii) analyse the relationships between match-day (MD) demands and wellness during the following day (MD+1); and (iv) analyse the relationships between MD and wellness during the day before match-play (MD-1). Thirteen professional soccer players (age: 24.85±3.13 years) were monitored daily over 16-weeks for wellness and training and match-play intensity. The daily wellness measures included fatigue, quality of sleep, muscle soreness, mood and stress using a 1-5 scale. Internal intensity was subjectively measured daily using the rating of perceived exertion (RPE) and the multiplication of RPE by session duration (s-RPE). While external intensity was quantified utilising high-speed running, sprinting, and acceleration and deceleration metrics. Data was analysed from each training session before (i.e., MD-5) or after the match (i.e., MD+1). Repeated measures ANOVA or Friedman ANOVA was used to analyse the aims (i) where Spearman correlation was applied to analyse the relationships between the aims (ii) and (iii) between sleep quality and training intensity. The main results for aim (i) showed that MD+1 presented the lowest values for wellness variables (p < 0.05). While MD-1 presented the lowest internal and external load values (for all variables), with MD presenting the highest values (p < 0.05). Regarding aim (ii), the main result showed significant large negative correlations between fatigue and s-RPE (r = -0.593; p = 0.033). Considering aim (iii), significant small to very large negative correlations were found for sleep quality, fatigue and muscle soreness with all internal and external variables (p < 0.05). Lastly, the main results for aim (iv) showed large negative correlations for fatigue and session duration; fatigue and s-RPE; muscle soreness and session duration; muscle soreness and s-RPE; and muscle soreness and decelerations (p < 0.05, for all). The main conclusions were that MD had an influence on wellness and internal and external training intensity, notably MD-1 and MD+1 were most affected. In this regard, a tendency of higher internal and external intensity on MD was associated with lower wellness measures of sleep quality, muscle soreness and fatigue on MD+1.


Insights into the diversity of NOD-like receptors: Identification and expression analysis of NLRC3, NLRC5 and NLRX1 in rainbow trout.
Nucleotide-binding oligomerization domain (NOD)-like receptors (NLRs) are efficient soluble intracellular sensors that activate defense mechanisms against pathogens. In teleost fish, the involvement of NLRs in the immune response is not well understood. However, recent work has evidenced the expression of different NLRs in response to some pathogen associated molecular patterns (PAMPs). In the present work, the cDNA sequence encoding three new NOD-like receptors were identified in Oncorhynchus mykiss, namely OmNLRC3, OmNLRC5 and OmNLRX1. Results showed that their sequences coded for proteins of 1135, 836 and 1010 amino acids, respectively. The deduced protein sequences of all receptors showed characteristic domains of this receptor family, such as leucine rich repeats and NACHT domain. Phylogenetic analysis revealed a high degree of identity with other NOD-like receptors and they are clustered into different families. Transcript expression analysis indicated that OmNLRs are constitutively expressed in liver, spleen, intestine, gill, skin and brain. OmNLR expression was upregulated in kidney and gills from rainbow trout in response to LPS. In order to give new insights into the function of these new NLR members, an in vitro model of immune stimulation was established using the rainbow trout cell line RTgill-W1. Expression analysis revealed that RTgill-W1 overexpressed proinflammatory cytokines in response to LPS and poly I:C alongside with a differential overexpression of OmNLRC3, OmNLRC5 and OmNLRX1. The expression of OmNLRC5 was further verified at the protein level by immunofluorescence. Finally, the effect of the overexpressed cytokines on the OmNLR expression by RTgill-W1 cells was assessed, suggesting a regulatory mechanism on OmNLRC3 expression. Overall, results suggest that O. mykiss NOD-like receptors could play a key role in the defense mechanisms of teleost through PAMP recognition. Future studies will focus on gills which could be related with a key sensor mucosal system in one of the most environmentally fish exposed tissues.


Coordinating actions.
The motor system composes complex actions by combining simpler submovements. This presumably involves sharing information about the progress of one submovement with the centres controlling another submovement, to ensure that the second happens in an appropriate relation to the first. This process is called coordination. In this paper I discuss evidence that coordinating actions indeed involves an active process of sharing information about the current state of movements. Coordination appears to be qualitatively different from the process of reacting to external stimuli. This may reflect the importance of predictive representations in coordination. Finally, the processes underlying coordination appear to be organized in a response-specific fashion, as a number of relatively independent circuits. The development and tuning of these circuits may, in part, be what makes an action "skilled".


Regulation of gene expression directed by the long terminal repeat of the feline immunodeficiency virus.
The long terminal repeat (LTR) of a retrovirus contains sequence elements that constitute a promoter for controlling viral gene expression in infected cells. We have examined regulation of LTR-directed gene expression in feline immunodeficiency virus (FIV), a T-lymphocytopathic lentivirus associated with a fatal AIDS-like disease in domestic cats. Two independent virus isolates, designated FIV-Petaluma and FIV-PPR, have been molecularly cloned and show greater than 85% sequence homology. Both clones (termed pF34 and pPPR) produce infectious virus after transfection of permissive feline cells. Basal promoter activity of the LTRs was measured in various cell lines in transient expression assays using plasmids containing the viral LTR linked to the bacterial chloramphenicol acetyltransferase gene. Both LTRs were strong promoters in several cell lines, although in some cell lines the pF34 LTR had four- to fivefold higher basal activity than the pPPR LTR. FIV LTR mutations affecting the first AP4 site, AP1 site, ATF site, or NF-kappa B site resulted in decreased basal activity of the FIV promoter. Mutational analysis also revealed a negative regulatory element. In cotransfection experiments, both pF34 proviral DNA and pPPR proviral DNA appeared to transactivate either the pF34 LTR or the pPPR LTR; however, levels of transactivation were very low. Cotransfection of both LTRs with FIV subgenomic clones containing various viral open reading frames resulted in low level or no transactivation. The LTRs of both FIV clones responded to cell activation signals in human T-lymphoid cells (Jurkat) treated with phytohemagglutinin and phorbol-12-myristate-13-acetate. Promoter function of both FIV LTRs was also enhanced in cells treated with either forskolin, an inducer of intracellular cyclic-AMP (c-AMP), or dibutyryl c-AMP. Analysis of site-specific mutants showed that a potential AP1 site in the U3 domain of the LTR was required for T-cell activation responses mediated by protein kinase C, whereas a putative ATF site was the target for c-AMP-induced responses mediated by protein kinase A. These studies revealed that cellular transcription factors play a significant role in regulation of FIV gene expression.


Clinical trial to evaluate immunogenicity and safety of inactivated hepatitis A vaccination starting at 2-month-old children.
Active immunization with hepatitis A vaccine has been shown to provide long-term protection against hepatitis A virus (HAV) infection. However, few data are available regarding use of the hepatitis A vaccine in children under two years of age. The present study was conducted to test the safety and immunogenicity of inactivated hepatitis A vaccine administered to infants, and to evaluate the correlation between mother and infant anti-HAV antibodies. A total of sixty healthy children, two months of age, were enrolled in this study and immunized with 360 EU of inactivated hepatitis A vaccine (Havrix) according to the two, four and six months of age schedule. Blood sampling was performed prior to the first vaccination and one month after the third vaccination at seven months. Venipuncture was also done on mother on admission. The reactogenicity was expressed as the percentage of reported local and systemic reactions. The most common side effects were erythema on the injection site and fever. Infants with passively transferred maternal anti-HAV antibodies had a reduced anti-HAV GMT after vaccination. On admission, only one infant and his mother were seronegative and seroconversion was only detected in this infant. One month after the third dose seven infants (12.3%) were found to be seronegative. The infant without passively acquired maternal anti-HAV had the protective levels with a GMT of 3176 mIU/ml one month following the third dose. There was a significant positive correlation between the titers of mother and infant anti-HAV antibodies (n = 0.96, p < 0.001) on admission. Hepatitis A vaccine showed no immunogenicity in infants with presence of maternal antibodies. Hepatitis A vaccine is safe but it should be used after the disappearance of maternal antibodies.


On the natural seeding of marshland pastures with bovine gastrointestinal parasites.
By placing parasite-free calves in paddocks grazed by infected animals for 18 day periods at various times during the previous season it was shown that eggs of Ostertagia ostertagi, Cooperia oncophora and Nematodirus helvetianus deposited on pastures from early July to October of one year were able to survive in the Maritime area of Canada over winter either as eggs and/or larvae and contribute to residual infections on these pastures the following spring. The greatest deposition and/or survival of those eggs that were shed on pasture occurred in August and September for Cooperia and in September and October for Ostertagia. Greatest deposition of Nematodirus occurred in July and August and relatively few Nematodirus eggs shed in late September or early October were infective early in the next season. The number of generations of worms per year was low, ranging from one to two or perhaps three per year depending on the species. There was a delay in the maturation of many worm eggs. Residual overwintering infections play a significant role in the establishment of initial infections each summer in susceptible stock. These animals recontaminate the pastures leading to the subsequent development of large numbers of infective larvae by late summer and autumn.


Evaluation of etiological hypotheses and a study of early lesions in alcoholic pancreatitis.
Current understanding of the pathology of chronic pancreatitis secondary to chronic alcoholism is exclusively based on surgical biopsies or autopsy studies of the pancreatic gland from far-advanced stages of the disease. Biopsy of the pancreas is dangerous, and good experimental models for alcoholic pancreatitis are not available to evaluate the chronology of events in the pancreas. A classification system was proposed many years ago with various hypotheses on the pathogenesis of chronic pancreatitis. These hypotheses will be examined in relation to the empirical evidence.


Crystal and molecular structure of DL-serine hydrochloride studied by X-ray diffraction, low-temperature Fourier transform infrared spectroscopy and DFT(B3LYP) calculations.
The structure of dl-serine.HCl was studied by three complementary techniques. Experimental Fourier transform infrared (FT-IR) spectra of pure NH/OH polycrystalline dl-serine.HCl [HO-CH2-CH(NH3+)-COOH.Cl(-)] and the respective deuterated derivatives [ND/ODAlcohol/Acid (<10% and ca. 60% D)] were recorded in the region 4000-400 cm(-1) in the temperature range 300-10 K and interpreted. The assignments were confirmed by comparison with the vibrational spectra of crystalline dl- and l-serine zwitterions [HO-CH 2-CH(NH3+)-COO(-)]. Further insight into the structure of the title compound was provided by theoretical DFT(B3LYP)/6-311++G(d,p) calculations of the infrared spectra and energies of 13 different conformers. Potential energy distributions resulting from normal co-ordinate analysis were calculated for the most stable conformer ( I) in its hydrogenated and deuterated modification. Frequencies of several vibrational modes were used in the estimation of enthalpies of individual H-bonds present in the crystal, using empirical correlations between enthalpy and the frequency shift that occurs as a result of the establishment of the H-bonds. X-ray crystallography data for dl-serine.HCl were recorded for the first time and, together with the experimental vibrational spectra and the theoretical calculations, allowed a detailed characterization of its molecular structure.