Drosophila increase exploration after visually detecting predators.
Novel stimuli elicit behaviors that are collectively known as specific exploration. These behaviors allow the animal to become more familiar with the novel objects within its environment. Specific exploration is frequently suppressed by defensive reactions to predator cues. Herein, we examine if this suppression occurs in Drosophila melanogaster by measuring the response of these flies to wild harvested predators. The flies used in our experiments have been cultured and had not lived under predator threat for multiple decades. In a circular arena with centrally-caged predators, wild type Drosophila actively avoided the pantropical jumping spider, Plexippus paykulli, and the Texas unicorn mantis, Phyllovates chlorophaena, indicating an innate defensive reaction to these predators. Interestingly, wild type Drosophila males also avoided a centrally-caged mock spider, and the avoidance of the mock spider became exaggerated when it was made to move within the cage. Visually impaired Drosophila failed to detect and avoid the Plexippus paykulli and the moving mock spider, while the broadly anosmic orco2 mutants were fully capable of detecting and avoiding Plexippus paykulli, indicating that these flies principally relied upon vison to perceive the predator stimuli. During early exploration of the arena, exploratory activity increased in the presence of Plexippus paykulli and the moving mock spider. The elevated activity induced by Plexippus paykulli disappeared after the fly had finished exploring, suggesting the flies were capable of habituating the predator cues. Taken together, these results indicate that despite being isolated from predators for decades Drosophila will visually detect these predators, retain innate defensive behaviors, respond by increasing exploratory activity in the arena rather than suppressing activity, and may habituate to normal predator cues.


Dr. Luigi Di Bella and the politics of elixirs.
Health Canada and the Canadian-Italian Physicians Association recently spent $20,000 to send 4 Canadian cancer specialists to Italy to check out the "miracle" anticancer cocktail prepared by an Italian physician. Most doctors remain decidedly sceptical about this latest "cure," but Charlotte Gray says their suspicions count for little when an issue like this captures the imagination of the public and their politicians.


The impact of menopause and serum DHEA-S level on the severity of Covid-19.
We aimed to demonstrate the difference between premenopausal and postmenopausal women in respect of the clinical course and outcomes of Covid-19. We investigated the epidemiological and hormonal factors which influence the severity of the disease. This observational cross-sectional study included the female patients admitted to a Covid-19 outpatient clinic between July 2020 and June 2021 and diagnosed with a positive polymerase chain reaction test. Blood samples were obtained to determine the serum levels of follicle stimulating hormone, luteinizing hormone, estradiol, total testosterone and dehydroepiandrosterone sulfate. Our primary outcome was the poor clinical course of the disease in postmenopausal women. Our secondary outcome was the contribution of hormonal status to the clinical course of the disease. Our cohort consisted of 253 female patients (85 with mild, 125 with moderate and 43 with severe disease; 101 at the premenopausal and 152 at the postmenopausal stage). There was a statistically significant difference between the patients in different severity groups regarding clinical data and serum levels of luteinizing hormone, follicle stimulating hormone, estradiol and dehydroepiandrosterone sulfate. Being one year younger decreased the odds of having severe Covid-19 0.338-fold relative to the group with mild disease. A decrease in the serum dehydroepiandrosterone sulfate level was associated with a 2.604-fold increase in the odds of having severe Covid-19 relative to the group with mild disease. Being postmenopausal increased the odds of having severe disease compared with mild disease by 2.687-fold. The prognosis of Covid-19 is more favorable in premenopausal women compared with postmenopausal women. Age, postmenopausal status and serum levels of dehydroepiandrosterone sulfate are important predictors of the severity of Covid-19 for women.


An optimized, chemically regulated gene expression system for Chlamydomonas.
Chlamydomonas reinhardtii is a model system for algal and cell biology and is used for biotechnological applications, such as molecular farming or biological hydrogen production. The Chlamydomonas metal-responsive CYC6 promoter is repressed by copper and induced by nickel ions. However, induction by nickel is weak in some strains, poorly reversible by chelating agents like EDTA, and causes, at high concentrations, toxicity side effects on Chlamydomonas growth. Removal of these bottlenecks will encourage the wide use of this promoter as a chemically regulated gene expression system. Using a codon-optimized Renilla luciferase as a reporter gene, we explored several strategies to improve the strength and reversibility of CYC6 promoter induction. Use of the first intron of the RBCS2 gene or of a modified TAP medium increases the strength of CYC6 induction up to 20-fold. In the modified medium, induction is also obtained after addition of specific copper chelators, like TETA. At low concentrations (up to 10 microM) TETA is a more efficient inducer than Ni, which becomes a very efficient inducer at higher concentrations (50 microM). Neither TETA nor Ni show toxicity effects at the concentrations used. Unlike induction by Ni, induction by TETA is completely reversible by micromolar copper concentrations, thus resulting in a transient "wave" in luciferase activity, which can be repeated in subsequent growth cycles. We have worked out a chemically regulated gene expression system that can be finely tuned to produce temporally controlled "waves" in gene expression. The use of cassettes containing the CYC6 promoter, and of modified growth media, is a reliable and economically sustainable system for the temporally controlled expression of foreign genes in Chlamydomonas.


The Effect of Upper Body Mass and Initial Knee Flexion on the Injury Outcome of Post Mortem Human Subject Pedestrian Isolated Legs.
In the ECE 127 Regulation on pedestrian leg protection, as well as in the Euro NCAP test protocol, a legform impactor hits the vehicle at the speed of 40 kph. In these tests, the knee is fully extended and the leg is not coupled to the upper body. However, the typical configuration of a pedestrian impact differs since the knee is flexed during most of the gait cycle and the hip joint applies an unknown force to the femur. This study aimed at investigating the influence of the inertia of the upper body (modelled using an upper body mass fixed at the proximal end of the femur) and the initial knee flexion angle on the lower limb injury outcome. In total, 18 tests were conducted on 18 legs from 9 Post Mortem Human Subjects (PMHS). The principle of these tests was to impact the leg at 40 kph using a sled equipped with 3 crushing steel tubes, the stiffness of which were representative of the front face of a European sedan (bonnet leading edge, bumper and spoiler). The mass of the equipped sled was 74.5 kg. The test matrix was designed to perform 4 tests in 4 configurations combining two upper body masses (either 0 or 3 kg) and two knee angles (0 or 20 degrees) at 40 kph (11 m/s) plus 2 tests at 9 m/s. Autopsies were performed on the lower limbs and an injury assessment was established. The findings of this study were first that the increase of the upper body mass resulted in more severe injuries, second that an initial flexion of the knee, corresponding to its natural position during the gait cycle, decreased the severity of the injuries, and third that based on the injury outcome, a test conducted with no upper body mass and the knee fully extended was as severe as a test conducted with a 3 kg upper body mass and an initial knee flexion of 20°.


Osteoblastoma of the mandible with root resorption: a case report.
This article reviews the clinical behavior, histologic features, differential diagnosis, and treatment of a benign osteoblastoma. Benign osteoblastoma is a rare tumor constituting 1% of all primary bone tumors. Only 15% of osteoblastomas occur in the skull and jaw bones. The most common clinical presentation is a painful or tender swelling. A case is presented of a 21-year-old female who had noted discomfort for approximately 2 years, and the pain was not relieved by any analgesic. The choice of treatment was local excision and curettage. In this case, root resorption of the adjacent tooth, which is not a characteristic behavior of osteoblastoma, is seen.


Interaction between 5-HTTLPR and 5-HT1B genotype status enhances cerebral 5-HT1A receptor binding.
Serotonergic neurotransmission is thought to underlie a dynamic interrelation between different key structures of the serotonin system. The serotonin transporter (SERT), which is responsible for the reuptake of serotonin from the synaptic cleft into the neuron, as well as the serotonin-1A (5-HT1A) and -1B (5-HT1B) receptors, inhibitory auto-receptors in the raphe region and projection areas, respectively, are likely to determine serotonin release. Thereby, they are involved in the regulation of extracellular serotonin concentrations and the extent of serotonergic effects in respective projection areas. Complex receptor interactions can be assessed in vivo with positron emission tomography (PET) and single-nucleotide-polymorphisms, which are thought to alter protein expression levels. Due to the complexity of the serotonergic system, gene × gene interactions are likely to regulate transporter and receptor expression and therefore subsequently serotonergic transmission. In this context, we measured 51 healthy subjects (mean age 45.5 ± 12.9, 38 female) with PET using [carbonyl-(11)C]WAY-100635 to determine 5-HT1A receptor binding potential (5-HT1A BPND). Genotyping for rs6296 (HTR1B) and 5-HTTLPR (SERT gene promoter polymorphism) was performed using DNA isolated from whole blood. Voxel-wise whole-brain ANOVA revealed a positive interaction effect of genotype groups (5-HTTLPR: LL, LS+SS and HTR1B: rs6296: CC, GC+GG) on 5-HT1A BPND with peak t-values in the bilateral parahippocampal gyrus. More specifically, highest 5-HT1A BPND was identified for individuals homozygous for both the L-allele of 5-HTTLPR and the C-allele of rs6296. This finding suggests that the interaction between two major serotonergic structures involved in serotonin release, specifically the SERT and 5-HT1B receptor, results in a modification of the inhibitory serotonergic tone mediated via 5-HT1A receptors.


Early hearing improvement predicts the prognosis of idiopathic sudden sensorineural hearing loss.
To determine pre- and post-treatment factors that are useful for predicting the prognosis of hearing improvement in idiopathic sudden sensorineural hearing loss (ISSHL). This retrospective study included 332 patients with ISSHL. Patients received intravenous steroid treatment (prednisolone sodium succinate; 120 mg/day followed by dose tapering). Complete recovery of hearing levels was defined as a final pure-tone audiometry of ≤ 20 dB HL or the same level as the contralateral ear. Patients' age; sex; side of hearing loss; initial hearing level; days from onset to treatment; presence of vertigo, diabetes, and hypertension; and hearing improvement on days 3-4 and 6-7 after treatment initiation were analyzed as potential prognostic factors. Overall, 109 patients (32%) had complete recovery. Results of the multivariate logistic regression model identified age (odds ratio [OR] = 0.974), initial hearing level (OR = 0.949), vertigo (OR = 0.409), and hearing improvement on days 6-7 after treatment initiation (OR = 1.11) as significant independent predictors of complete recovery. Age ≥ 60 years, initial hearing level ≥ 72.5 dB HL, and vertigo contributed to poor prognosis. Patients without these three factors and a hearing improvement of ≥ 10 dB HL on days 6-7 post-treatment had a complete recovery rate of 80%. Only 1.5% of the patients with 2-3 of these factors and a hearing improvement of < 10 dB HL on days 6-7 after treatment initiation achieved complete recovery. Age, initial hearing level, vertigo, and hearing improvement on days 6-7 after treatment initiation were independent predictors of hearing recovery in ISSHL.


Geographic variation in adaptation at the molecular level: a case study of plant immunity genes.
Natural selection imposed by interacting species frequently varies among geographic locations and can lead to local adaptation, where alternative phenotypes are found in different populations. Little is known, however, about whether geographically variable selection acting on traits that mediate species interactions is consistent or strong enough to influence patterns of nucleotide variation at individual loci. To investigate this question, we examined patterns of nucleotide diversity and population structure at 16 plant innate immunity genes, with putative functions in defending plants against pathogens or herbivores, from six populations of teosinte (Zea mays ssp. parviglumis). Specifically, we tested whether patterns of population structure and within-population diversity at immunity genes differed from patterns found at nonimmunity (reference) loci and from neutral expectations derived from coalescent simulations of structured populations. For the majority of genes, we detected no strong evidence of geographically variable selection. However, in the wound-induced serine protease inhibitor (wip1), which inhibits the hydrolysis of dietary proteins in insect herbivores, one population showed unusually high levels of genetic differentiation, very low levels of nucleotide polymorphism, and was fixed for a novel replacement substitution in the active site of the protein. Taken together, these data suggest that wip1 experienced a recent selective sweep in one geographic region; this pattern may reflect local adaptation or an ongoing species-wide sweep. Overall, our results indicate that a signature of local adaptation at the molecular level may be uncommon-particularly for traits that are under complex genetic control.


A brief period of sleep deprivation leads to subtle changes in mouse gut microbiota.
Not getting enough sleep is a common problem in our society and contributes to numerous health problems, including high blood pressure, diabetes and obesity. Related to these observations, a wealth of studies has underscored the negative impact of both acute and chronic sleep deprivation on cognitive function. More recently it has become apparent that the gut microbiota composition can be rapidly altered, modulates brain function and is affected by the aforementioned health problems. As such, changes in the microbiota composition may contribute to the behavioural and physiological phenotypes associated with sleep deprivation. It is unclear, however, whether a brief period of sleep deprivation can also negatively impact the gut microbiota. Here, we examined the impact of 5 hr of sleep deprivation on gut microbiota composition of male C57Bl6/J mice. Despite the fact that the overall microbial composition did not change between the control- and sleep-deprived groups, the relative abundance of the Clostridiaceae and Lachnospiraceae were slightly altered in sleep-deprived animals compared to controls. Together, these data suggest that depriving mice of sleep for 5 hr leads to subtle changes in the gut microbiota composition.


Potential molecular basis of different physiological properties of the transient outward K+ current in rabbit and human atrial myocytes.
The properties of the transient outward current (Ito) differ between rabbit and human atrial myocytes. In particular, rabbit Ito is known to recover more slowly than its human counterpart and to show much more frequency dependence. To assess the possibility that these physiological differences may reflect differing expression of K+ channel subunit gene products, we used a combination of whole-cell voltage-clamp, heterologous expression, pharmacological, antisense, and Western blot techniques. The inactivation of Ito in rabbit atrial myocytes was significantly slowed by hydrogen peroxide, with human Ito being unaffected. Use-dependent unblocking with 4-aminopyridine was not seen for rabbit Ito nor for Kv1.4 currents in Xenopus oocytes, whereas human Ito showed strong use-dependent unblock (as did Kv4 currents). Western blots indicated the presence of Kv4 proteins in both human and rabbit atrial membranes, but Kv1.4 was only detected in the rabbit. Antisense oligodeoxynucleotides directed against Kv4.3, Kv4.2, or Kv1.4 subunit sequences significantly inhibited Ito current density in cultured rabbit atrial myocytes, whereas only Kv4.3 antisense significantly inhibited Ito in human cells. Neither mismatch oligodeoxynucleotides nor vehicle altered currents in either species. We conclude that, unlike human atrial myocytes, rabbit atrial myocytes express Kv1.4 channel subunits, which likely contribute to a number of important physiological differences in Ito properties between the species. To our knowledge, these studies constitute the first demonstration of a functional role for Kv1.4 channels in cardiac membranes and provide insights into the molecular mechanisms of an important cardiac repolarizing current.


Use and Effectiveness of Sleep Medications by Parent Report in Individuals with Williams Syndrome.
Sleep disorders are common in individuals with Williams syndrome (WS), and sleep disturbance has a significant negative effect on attention, learning, and behavior. The use of sleep-inducing medicine in individuals with WS has not been widely investigated. The objective of this study was to evaluate the use and effectiveness of sleep medications among a large sample of individuals with WS by parent survey. A survey of the use and effectiveness of sleep medications was completed by 513 (of 2846) members of the Williams Syndrome Association. The online survey asked for the age at initiation, degree of effectiveness (helpful, somewhat helpful, and not helpful), and side effects. One hundred thirty participants (25%) indicated that their family member with WS had taken medication to help with sleep. Melatonin was the most commonly reported medication taken for sleep, with 91% of parents reporting that it was "helpful" or "somewhat helpful" for their child with WS. Those who reported taking melatonin reported very few, if any, side effects. This parent-completed survey is a preliminary study showing the positive benefit of melatonin for individuals with WS who have disrupted sleep. The findings support the need for further study of the use of melatonin, in addition to behavioral sleep aids, given evidence that sleep disturbance negatively influences cognition and behavior.


[Organization of centromere regions of chromosomes in the lampbrush phase].
Centromeres have a pivotal role in the complex of structural elements that are required for precise segregation of eukaryotic chromosomes during two types of cellular divisions--mitosis and meiosis. Data of ultrastructural and cytomolecular analysis indicate significant changes in molecular composition and functional morphology of centromeres during preparation for the first meiotic division. The review is devoted to modern concepts of morpho-functional organization of chromosomal centromere regions in growing oocytes in birds and amphibians. Structure, molecular composition as well as domain organization of centromeres in the lampbrush phase are characterized; data of cytogenetic analysis are presented. Special attention is given to the significance and regulation of satellite DNA transcription in the nuclei of developing oocytes. Possible functions of centromere "protein bodies" formed at the primary constriction of meiotic bivalents are discussed.


Mechanosensory neurons control sweet sensing in Drosophila.
Animals discriminate nutritious food from toxic substances using their sense of taste. Since taste perception requires taste receptor cells to come into contact with water-soluble chemicals, it is a form of contact chemosensation. Concurrent with that contact, mechanosensitive cells detect the texture of food and also contribute to the regulation of feeding. Little is known, however, about the extent to which chemosensitive and mechanosensitive circuits interact. Here, we show Drosophila prefers soft food at the expense of sweetness and that this preference requires labellar mechanosensory neurons (MNs) and the mechanosensory channel Nanchung. Activation of these labellar MNs causes GABAergic inhibition of sweet-sensing gustatory receptor neurons, reducing the perceived intensity of a sweet stimulus. These findings expand our understanding of the ways different sensory modalities cooperate to shape animal behaviour.


A conserved acidic residue in phenylalanine hydroxylase contributes to cofactor affinity and catalysis.
The catalytic domains of aromatic amino acid hydroxylases (AAAHs) contain a non-heme iron coordinated to a 2-His-1-carboxylate facial triad and two water molecules. Asp139 from Chromobacterium violaceum PAH (cPAH) resides within the second coordination sphere and contributes key hydrogen bonds with three active site waters that mediate its interaction with an oxidized form of the cofactor, 7,8-dihydro-l-biopterin, in crystal structures. To determine the catalytic role of this residue, various point mutants were prepared and characterized. Our isothermal titration calorimetry (ITC) analysis of iron binding implies that polarity at position 139 is not the sole criterion for metal affinity, as binding studies with D139E suggest that the size of the amino acid side chain also appears to be important. High-resolution crystal structures of the mutants reveal that Asp139 may not be essential for holding the bridging water molecules together, because many of these waters are retained even in the Ala mutant. However, interactions via the bridging waters contribute to cofactor binding at the active site, interactions for which charge of the residue is important, as the D139N mutant shows a 5-fold decrease in its affinity for pterin as revealed by ITC (compared to a 16-fold loss of affinity in the case of the Ala mutant). The Asn and Ala mutants show a much more pronounced defect in their kcat values, with nearly 16- and 100-fold changes relative to that of the wild type, respectively, indicating a substantial role of this residue in stabilization of the transition state by aligning the cofactor in a productive orientation, most likely through direct binding with the cofactor, supported by data from molecular dynamics simulations of the complexes. Our results indicate that the intervening water structure between the cofactor and the acidic residue masks direct interaction between the two, possibly to prevent uncoupled hydroxylation of the cofactor before the arrival of phenylalanine. It thus appears that the second-coordination sphere Asp residue in cPAH, and, by extrapolation, the equivalent residue in other AAAHs, plays a role in fine-tuning pterin affinity in the ground state via deformable interactions with bridging waters and assumes a more significant role in the transition state by aligning the cofactor through direct hydrogen bonding.


A relapse of POEMS syndrome presenting as acute kidney injury.
Peripheral neuropathy, organomegaly, endocrinopathy, monoclonal protein and skin changes (POEMS) syndrome is a rare disease, and only in a minority of cases, causes an impairment of kidney function. Here, we describe a case of a 55-year-old man with a history of POEMS syndrome who presented with acute kidney injury following a routine blood test. On further investigation, a relapse in POEMS syndrome was diagnosed, uniquely isolated to renal involvement.


Threshold Concepts in a Literary Magazine Produced by Health Care Students.
This article was migrated. The article was marked as recommended. Introduction: Progress Notes is a literary magazine featuring works from health care students in the USA. It is edited by medical students at the Uniformed Services University of the Health Sciences (USUHS) to provide a space for their creative works. Since the magazine's inception in 2016, four volumes have been published, containing 130 works of poetry, fiction, reflection, and artwork. Goal: To understand the themes and perspectives of the published works. Methods: After Institutional Review Board approval, two researchers used qualitative thematic analysis to examine the texts, coding independently and resolving differences by discussion. They arranged the codes into themes which were discussed until consensus. A third researcher read the texts, codes and themes and verified that they were an accurate reflection of the published works. Artwork was assigned themes by the same two researchers who analyzed the written works, and a sample selection was verified by the third researcher. Results: Researchers identified eight themes across poetry, fiction, and reflective essays: vocation, death, failure/resilience, emotional restraint, personhood of the patient, approach to the patient, military physicians, and moments of personal realization. Four themes were identified in the artwork: death; comradeship/aloneness; vocation/quest; and competence. Discussion: Students submitted creative works in which they grapple with what it means to be a physician. Analyzed through the lens of the threshold concepts, researchers identified: "I am a healer;" "I can deal with ambiguity;" "The patient is the focus;" "As a military medical officer, I serve two masters;" and "As a physician, I have a unique and complex relationship with death." These threshold concepts represent an ontological shift in the students' professional identity. Conclusion: A literary magazine edited, and featuring works by, health care students provides a forum in which health care students wrestle with the elusive and enigmatic fundamental principles of being a physician.


Implementing an advanced waste separation step in an MBT plant: assessment of technical, economic and environmental impacts.
Heavy fractions resulting from mechanical treatment stages of mechanical-biological waste treatment plants are posing very specific demands with regard to further treatment (large portions of inert and high-caloric components). Based on the current Austrian legal situation such a waste stream cannot be landfilled and must be thermally treated. The aim of this research was to evaluate if an inert fraction generated from this waste stream with advanced separation technologies, two sensor-based [near-infrared spectroscopy (NIR), X-ray transmission (XRT)] and two mechanical systems (wet and dry) is able to be disposed of. The performance of the treatment options for separation was evaluated by characterizing the resulting product streams with respect to purity and yield. Complementing the technical evaluation of the processing options, an assessment of the economic and global warming effects of the change in waste stream routing was conducted. The separated inert fraction was evaluated with regard to landfilling. The remaining high-caloric product stream was evaluated with regard to thermal utilization. The results show that, in principal, the selected treatment technologies can be used to separate high-caloric from inert components. Limitations were identified with regard to the product qualities achieved, as well as to the economic expedience of the treatment options. One of the sensor-based sorting systems (X-ray) was able to produce the highest amount of disposeable heavy fraction (44.1%), while having the lowest content of organic (2.0% C biogenic per kg waste input) components. None of the high-caloric product streams complied with the requirements for solid recovered fuels as defined in the Austrian Ordinance on Waste Incineration. The economic evaluation illustrates the highest specific treatment costs for the XRT (€ 23.15 per t), followed by the NIR-based sorting system (€ 15.67 per t), and the lowest costs for the air separation system (€ 10.79 per t). Within the ecological evaluation it can be shown that the results depend strongly on the higher heating value of the high caloric light fraction and on the content of C biogenic of the heavy fraction. Therefore, the XRT system had the best results for the overall GWP [-14 kg carbon dioxide equivalents (CO2 eq) per t of input waste] and the NIR-based the worst (193 kg CO2 eq per t of input waste). It is concluded that three of the treatment options would be suitable under the specific conditions considered here. Of these, sensor-based sorting is preferable owing to its flexibility.


Additional predisposing risk factors for atherothrombotic cerebrovascular disease among treated hypertensive volunteers.
A 7-year prospective study of a cohort of 107 neurologically normal elderly hypertensive volunteers (mean age, 65.8 +/- 8.3 years) was undertaken to investigate the predictive validities of additional risk factors for atherothrombotic cerebrovascular disease including stroke, transient ischemic attacks, reversible ischemic neurological deficits, and multi-infarct dementia. This longitudinal study has been in progress now for 7 years with a mean follow-up interval of 50.12 +/- 5.76 months. Among 107 formerly symptom-free, normal hypertensive volunteers, 25 (23%) have developed cerebrovascular disease, 7 (6.5%) sustained a stroke, 10 (9.3%) developed multi-infarct dementia, and 18 (16.8%) have transient ischemic attacks. None have suffered intracranial hemorrhage. Mean gray matter cerebral blood flow (CBF) values measured at the initial visit were sensitive predictors of cerebrovascular disease. Eight of 16 hypertensives (50%) with initial CBF values below 60.0 ml/100 g/min now exhibit signs and symptoms of cerebrovascular disease, while 11 of 43 hypertensives (25.6%) with initial CBF values between 60.1 and 69.9 ml/100 g/min and only 6 of 48 (12.5%) with initial CBF levels above 70.0 developed cerebrovascular disease. Incidence of cerebrovascular disease among cigarette smoking hypertensive volunteers (32.5%) was significantly greater than among nonsmokers (17.2%).


[Comparison of two psychophysical methods for the estimation of the severity of illness].
In this study, 15 diagnoses were selected and evaluated by the methods of estimation of magnitudes and estimation of categories. These methods were performed by 46 subjects (10 physicians, 20 nurses and 16 psychologists). The results show that the non-metric continuum of seriousness of an illness exhibits prosthetic characteristics and the variability of estimates is a linear function of the magnitude estimation, thus confirming that Ekman's law is also valid for non-metric continua.


Neuroretinal dysfunction with intact blood-retinal barrier and absent vasculopathy in type 1 diabetes.
It is unknown whether independent neural damage may occur in the pre-/absent vascular diabetic retinopathy (DR). To exclude vasculopathy, it is important to measure the integrity of the blood-retinal barrier (BRB). This cross-sectional study addressed this problem in type 1 diabetic patients with normal ocular fundus and absent breakdown of the BRB (confirmed with vitreous fluorometry). These were compared with a group with disrupted BRB (with normal fundus or initial DR) and normal controls. Multifocal electroretinography and chromatic/achromatic contrast sensitivity were measured in these 42 patients with preserved visual acuity. Amplitudes of neurophysiological responses (multifocal electroretinogram) were decreased in all eccentricity rings in both clinical groups, when compared with controls, with sensitivity >78% for a specificity level of 90%. Implicit time changes were also found in the absence of initial DR. Impaired contrast sensitivity along chromatic axes was also observed, and achromatic thresholds were also different between controls and both clinical groups. The pattern of changes in the group without baseline BRB permeability alterations, as probed by psychophysical and electrophysiological measurements, does thereby confirm independent damage mechanisms. We conclude that retinal neuronal changes can be diagnosed in type 1 diabetes, independently of the breakdown of the BRB and onset of vasculopathy.


Inhibition of COX-2, mPGES-1 and CYP4A by isoliquiritigenin blocks the angiogenic Akt signaling in glioma through ceRNA effect of miR-194-5p and lncRNA NEAT1.
Arachidonic acid (AA) metabolic enzymes including cyclooxygenase-2 (COX-2), microsomal prostaglandin E synthase-1 (mPGES-1) and cytochrome P450 (CYP) 4A11 play important roles in glioma angiogenesis. Thus, there is an urgent need to identify the underlying mechanisms and develop strategies to overcome them. A homology model of human CYP4A11 was constructed using SYBYL-X 2.0. Structure-based virtual screening against COX-2, mPGES-1 and CYP4A11was performed using the Surflex-Dock of the SYBYL suite. The candidates were further evaluated their antiangiogenic activities in a zebrafish embryo and rabbit corneal angiogenesis model. Laser doppler analysis was used to measure tumor perfusion. The expression of CD31 and α-SMA was measured by immunofluorescence. Western blot was used to measure the expression of HIF-1, Akt and p-Akt. The gene expression of FGF-2, G-CSF, PDGF, TGF-β, Tie-2, VEGF, lncRNA NEAT1 and miR-194-5p were determined using qPCR. The production of FGF-2, TGF-β and VEGF were analyzed using ELISA. Bioinformatic analysis and luciferase reporter assays confirmed the interaction between lncRNA NEAT1 and miR-194-5p. The nearly 36,043 compounds from the Traditional Chinese Medicine (TCM) database were screened against COX-2, mPGES-1 and CYP4A11 3D models, and the 17 top flavonoids were identified. In zebrafish screening, isoliquiritigenin (ISL) exhibited the most potent antiangiogenic activities with the EC50 values of 5.9 μM. Conversely, the antiangiogenic effects of ISL in the zebrafish and rabbit corneal models were partly reversed by 20-hydroxyeicosatetraenoic acid (20-HETE) or prostaglandin E2 (PGE2). ISL normalized glioma vasculature and improved the efficacy of temozolomide therapy in the rat C6 glioma model. Inhibition of COX-2, mPGES-1 and CYP4A by ISL decreased FGF-2, TGF-β and VEGF production in the C6 and U87 glioma cells with p-Akt downregulation, which was reversed by Akt overexpression. Furthermore, ISL downregulated lncRNA NEAT1 but upregulated miR-194-5p in the U87 glioma cell. Importantly, lncRNA NEAT1 overexpression reversed ISL-mediated increase in miR-194-5p expression, and thereby attenuated FGF-2, TGF-β and VEGF production. Reprogramming COX-2, mPGES-1 and CYP4A mediated-AA metabolism in glioma by flavonoid ISL inhibits the angiogenic Akt- FGF-2/TGF-β/VEGF signaling through ceRNA effect of miR-194-5p and lncRNA NEAT1, and may serve as a novel therapeutic strategy for human glioma.


Converting beach-cast seagrass wrack into biochar: A climate-friendly solution to a coastal problem.
Excessive accumulation of plant 'wrack' on beaches as a result of coastal development and beach modification (e.g. groin installation) is a global problem. This study investigated the potential for converting beach-cast seagrass wrack into biochar as a 'climate-friendly' disposal option for resource managers. Wrack samples from 11 seagrass species around Australia were initially screened for their biochar potential using pyrolysis techniques, and then two species - Posidonia australis and Zostera muelleri - underwent detailed analyses. Both species had high levels of refractory materials and high conversion efficiency (48-57%) of plant carbon into biochar carbon, which is comparable to high-quality terrestrial biochar products. P. australis wrack gave higher biochar yields than Z. muelleri consistent with its higher initial carbon content. According to 13C NMR, wrack predominantly comprised carbohydrates, protein, and lignin. Aryl carbon typical of pyrogenic materials dominated the spectrum of the thermally-altered organic materials. Overall, this study provides the first data on the feasibility of generating biochar from seagrass wrack, showing that biocharring offers a promising climate-friendly alternative to disposal of beach wrack in landfill by avoiding a portion of the greenhouse gas emissions that would otherwise occur if wrack was left to decompose.


Antibiotic resistance in acne: changes, consequences and concerns.
Antibiotic resistance in acne was first observed in the 1970s and has been a major concern in dermatology since the 1980s. The resistance rates and types of antimicrobials have subsequently shown great variations in regions and countries. Illustrative of this is the resistance to topical erythromycin and clindamycin which continues to be a problem worldwide, while resistance to systemic treatment with tetracyclines has remained low during the past decade. The resistance for the newer macrolides like azithromycin and clarithromycin has been increasing. The results of antibiotic resistance may include treatment failure of acne, disturbance of skin microbiota, induction of opportunistic pathogens locally and systemically, and dissemination of resistant strains to both healthcare personnel and the general population. The ensuing complications, such as aggravated opportunistic infections caused by Propionibacterium acnes and the emergence of multiresistant superbugs, have not yet been confirmed.


[Delirium in the clinical practice of a therapist].
Delirium is a neuropsychiatric condition that may complicate any visceral disease. Its rate is especially high among patients with inflammatory diseases or metabolic disturbances and in the elderly. Brain injury concurrent with an abnormal stress response underlies the development of delirium. The clinical picture of delirium is characterized by clouding of consciousness accompanied by global cognitive and behavioral changes. According to the nature of changes in motor behavior, delirium is divided into hyperactive, hypoactive, and mixed subtypes. Special scales, such as Confusion Assessment Method for the Intensive Care Unit (CAM-ICU), are used to identify delirium. Management of delirium includes specific therapy for the underlying disease and adequate care. Low-dose neuroleptics, haloperidol in particular, are recommended to correct behavioral changes.


How can malaria rapid diagnostic tests achieve their potential? A qualitative study of a trial at health facilities in Ghana.
Rapid diagnostic tests (RDTs) for malaria are at the early stages of introduction across malaria endemic countries. This is central to efforts to decrease malaria overdiagnosis and the consequent overuse of valuable anti-malarials and underdiagnosis of alternative causes of fever. Evidence of the effect of introducing RDTs on the overprescription of anti-malarials is mixed. A recent trial in rural health facilities in Ghana reduced overprescription of anti-malarials, but found that 45.5% patients who tested negative with RDTs were still prescribed an anti-malarial. A qualitative study of this trial was conducted, using in-depth interviews with a purposive sample of health workers involved in the trial, ranging from those who continued to prescribe anti-malarials to most patients with negative RDT results to those who largely restricted anti-malarials to patients with positive RDT results. Interviews explored the experiences of using RDTs and their results amongst trial participants. Meanings of RDTs were constructed by health workers through participation with the tests themselves as well as through interactions with colleagues, patients and the research team. These different modes of participation with the tests and their results led to a change in practice for some health workers, and reinforced existing practice for others. Many of the characteristics of RDTs were found to be inherently conducive to change, but the limited support from purveyors, lack of system antecedents for change and limited system readiness for change were apparent in the analysis. When introduced with a limited supporting package, RDTs were variously interpreted and used, reflecting how health workers had learnt how to use RDT results through participation. To build confidence of health workers in the face of negative RDT results, a supporting package should include local preparation for the innovation; unambiguous guidelines; training in alternative causes of disease; regular support for health workers to meet as communities of practice; interventions that address negotiation of health worker-patient relationships and encourage self-reflection of practice; feedback systems for results of quality control of RDTs; feedback systems of the results of their practice with RDTs; and RDT augmentation such as a technical and/or clinical troubleshooting resource.


[Effect of fasting on the content and transport of potassium ions in rat liver mitochondria].
The paper deals with the effect of fasting on the content and transport of potassium ions in the rat liver mitochondria as well as on oxidative phosphorylation. It is shown that the concentration of potassium ions in the mitochondria under fasting lowers by 20-30% at an average as compared to the control. The rate of potassium ions release from the mitochondria in the presence of DNP also lowers. It is found that the respiratory control ratio after 24-h fasting decreases and increases after 48-h fasting. Possible reasons of changes in the transport of potassium ions in the rat liver mitochondria in fasting are discussed.


A near-infrared light-controlled, ultrasensitive one-step photoelectrochemical detection of dual cell apoptosis indicators in living cancer cells.
Here, a near-infrared (NIR) light-controlled, ultrasensitive one-step photoelectrochemical (PEC) strategy was constructed to simultaneously detect cell apoptosis indicators, phosphatidylserine (Pho) and sodium-potassium adenosine triphosphatase (Sat), on living cancer cells. Using NIR light as excitation, the signal probe methylene blue (Tagkinetic) could be released, leading to a gradually decreased photocurrent signal Ikinetic; meanwhile, the photocurrent Istable of the signal probe carbon quantum dots (Tagstable) remained stable. The simultaneous detection of Pho and Sat could be achieved based on rapid one-step PEC detection under single NIR light with the assistance of a smart signal decryption strategy with Ikinetic and Istable. Importantly, this proposal provides more effective drug candidates with milder pharmaceutical effect but improved safety.


Imaging Reversible Mitochondrial Membrane Potential Dynamics with a Masked Rhodamine Voltage Reporter.
Mitochondria are the site of aerobic respiration, producing ATP via oxidative phosphorylation as protons flow down their electrochemical gradient through ATP synthase. This negative membrane potential across the inner mitochondrial membrane (ΔΨm) represents a fundamental biophysical parameter central to cellular life. Traditional, electrode-based methods for recording membrane potential are impossible to implement on mitochondria within intact cells. Fluorescent ΔΨm indicators based on cationic, lipophilic dyes are a common alternative, but these indicators are complicated by concentration-dependent artifacts and the requirement to maintain dye in the extracellular solution to visualize reversible ΔΨm dynamics. Here, we report the first example of a fluorescent ΔΨm reporter that does not rely on ΔΨm-dependent accumulation. We redirected the localization of a photoinduced electron transfer (PeT)-based indicator, Rhodamine Voltage Reporter (RhoVR), to mitochondria by masking the carboxylate of RhoVR 1 as an acetoxymethyl (AM) ester. Once within mitochondria, esterases remove the AM ester, trapping RhoVR inside of the mitochondrial matrix, where it can incorporate within the inner membrane and reversibly report on changes in ΔΨm. We show that this Small molecule, Permeable, Internally Redistributing for Inner membrane Targeting Rhodamine Voltage Reporter, or SPIRIT RhoVR, localizes to mitochondria across a number of different cell lines and responds reversibly to changes in ΔΨm induced by exceptionally low concentrations of the uncoupler FCCP without the need for exogenous pools of dye (unlike traditional, accumulation-based rhodamine esters). SPIRIT RhoVR is compatible with multi-color imaging, enabling simultaneous, real-time observation of cytosolic Ca2+, plasma membrane potential, and reversible ΔΨm dynamics.


[Dmt1]N/OFQ(1-13)-NH2: a potent nociceptin/orphanin FQ and opioid receptor universal agonist.
Intrathecally (i.t.) administered nociceptin/orphanin FQ (N/OFQ) evokes antinociceptive effects in rodents. Recent studies in monkeys demonstrated that i.t. co-application of N/OFQ and morphine elicits synergistic antinociceptive actions suggesting mixed N/OFQ peptide (NOP) and μ opioid receptor agonists as innovative spinal analgesics. Thus, novel N/OFQ related peptides were synthesized in order to identify and pharmacologically characterize a mixed NOP/ μ opioid receptor agonist. The following in vitro assays were used: calcium mobilization in cells expressing the human NOP or classical opioid receptors and chimeric G proteins, receptor and [(35)S]-GTPγS binding, [(35)S]-GTPγS binding in rat spinal cord membranes, guinea pig ileum bioassay. In vivo experiments were performed in monkeys using the tail withdrawal assay. From calcium mobilization studies [Dmt(1)]N/OFQ(1-13)-NH(2) was selected as the most potent and least selective compound. The mixed NOP/opioid full agonist activity and high affinity of [Dmt(1)]N/OFQ(1-13)-NH(2) was confirmed at human recombinant receptors in receptor binding, calcium mobilization and/or [(35)S]-GTPγS binding studies, at rat spinal cord receptors in [(35)S]-GTPγS binding experiments, and at guinea pig receptors inhibiting neurogenic contractions in the ileum. In vivo in the tail withdrawal assay in monkeys i.t. [Dmt(1) ]N/OFQ(1-13)-NH(2) was able to elicit robust and long-lasting antinociceptive effects. Collectively, these results demonstrate that [Dmt(1)]N/OFQ(1-13)-NH(2) behaves as NOP/opioid receptor universal agonist and substantiate the suggestion that such mixed ligands are worthy of development as innovative spinal analgesics.


Role of lipid A palmitoylation in bacterial pathogenesis.
The presence of palmitate in a minor fraction of lipid A has been known since the chemical structure of lipid A was first elucidated, but the functional importance in bacterial pathogenesis of regulated lipid A palmitoylation has become clear only recently. A palmitate chain from a phospholipid is incorporated into lipid A by an outer membrane enzyme PagP. The isolation of pagP mutants from pathogenic Gram-negative bacteria has revealed that palmitoylated lipid A can both protect the bacterium from certain host immune defenses and attenuate the ability of lipid A to activate those same defenses through the TLR4 signal transduction pathway. The mechanisms by which bacteria regulate the incorporation of palmitate into lipid A strikingly reflect the corresponding organism's pathogenic lifestyle. Variations on these themes can be illustrated with the known pagP homologs from Gram-negative bacteria, which include pathogens of humans and other mammals in addition to pathogens of insects and plants. The PagP enzyme is now lending itself both as a target for the development of anti-infective agents, and as a tool for the synthesis of lipid A-based vaccine adjuvants and endotoxin antagonists.


Alpinia oxyphylla Miq. fruit extract activates IGFR-PI3K/Akt signaling to induce Schwann cell proliferation and sciatic nerve regeneration.
It is known that the medicinal herb Alpinia oxyphylla Miq. is widely used as a remedy for diarrhea as well as the symptoms accompanying hypertension and cerebrovascular disorders. Moreover, it has also been reported that Alpinia oxyphylla Miq. has beneficial effects on anti-senescence and neuro-protection. This study focuses on the molecular mechanisms by which the Alpinia oxyphylla Miq. fruits promote neuron regeneration. A piece of silicone rubber was guided across a 15 mm gap in the sciatic nerve of a rat. This nerve gap was then filled with various doses of Alpinia oxyphylla Miq. fruits to assess their regenerative effect on damaged nerves. Further, we investigated the role of Alpinia oxyphylla Miq. fruits in RSC96 Schwann cell proliferation. Our current results showed that treatment with the extract of Alpinia oxyphylla Miq. fruits triggers the phosphorylated insulin-like growth factor-1 receptor- phosphatidylinositol 3-kinase/serine-threonine kinase pathway, and up-regulated the proliferating cell nuclear antigen in a dose-dependent manner. Cell cycle analysis on RSC96 Schwann cells showed that, after exposure to Alpinia oxyphylla Miq. fruit extract, the transition from the first gap phase to the synthesis phase occurs in 12-18 h. The expression of the cell cycle regulatory proteins cyclin D1, cyclin E and cyclin A increased in a dose-dependent manner. Transfection with a small interfering RNA blocked the expression of phosphatidylinositol 3-kinase and induced down-regulation both on the mRNA and protein levels, which resulted in a reduction of the expression of the survival factor B-cell lymphoma 2. We provide positive results that demonstrate that Alpinia oxyphylla Miq. fruits facilitate the survival and proliferation of RSC96 cells via insulin-like growth factor-1 signaling.


Are antioxidants helpful for disease prevention?
Free radicals are produced continuously in the cells as part of normal cellular function, however excess production might play a role in pathophysiology of many disease conditions, including cancer, Alzheimer's disease, atherosclerosis and some of the drug-induced toxicity. Many basic research studies and observational epidemiologic studies in human suggest that antioxidants can prevent oxidative damage. However, this is still a controversial issue because the results of clinical trials have been inconsistent. This article provides a brief overview of some of the diseases which are associated with free radicals, then discusses the roles of some of dietary antioxidant supplements in disease prevention, with particular reference to the findings of latest clinical trials.


Hypoxemia in School-age Children Undergoing One-lung Ventilation: A Retrospective Cohort Study from the Multicenter Perioperative Outcomes Group.
Risk factors for hypoxemia in school-age children undergoing one-lung ventilation remain poorly understood. The hypothesis was that certain modifiable and nonmodifiable factors may be associated with increased risk of hypoxemia in school-age children undergoing one-lung ventilation and thoracic surgery. The Multicenter Perioperative Outcomes Group database was queried for children 4 to 17 yr of age undergoing one-lung ventilation. Patients undergoing vascular or cardiac procedures were excluded. The original cohort was divided into two cohorts: 4 to 9 and 10 to 17 yr of age inclusive. All records were reviewed electronically for the primary outcome of hypoxemia during one-lung ventilation, which was defined as an oxygen saturation measured by pulse oximetry (Spo2) less than 90% for 3 min or longer continuously, while severe hypoxemia was defined as Spo2 less than 90% for 5 min or longer. Potential modifiable and nonmodifiable risk factors associated with these outcomes were evaluated using separate multivariable least absolute shrinkage and selection operator regression analyses for each cohort. The covariates evaluated included age, extremes of weight, American Society of Anesthesiologists Physical Status of III or higher, duration of one-lung ventilation, preoperative Spo2 less than 98%, approach to one-lung ventilation, right operative side, video-assisted thoracoscopic surgery, lower tidal volume ventilation (defined as tidal volume of 6 ml/kg or less and positive end-expiratory pressure of 4 cm H2O or greater for more than 80% of the duration of one-lung ventilation), and procedure type. The prevalence of hypoxemia in the 4- to 9-yr-old cohort and the 10- to 17-yr-old cohort was 24 of 228 (10.5% [95% CI, 6.5 to 14.5%]) and 76 of 1,012 (7.5% [95% CI, 5.9 to 9.1%]), respectively. The prevalence of severe hypoxemia in both cohorts was 14 of 228 (6.1% [95% CI, 3.0 to 9.3%]) and 47 of 1,012 (4.6% [95% CI, 3.3 to 5.8%]). Initial Spo2 less than 98% was associated with hypoxemia in the 4- to 9-yr-old cohort (odds ratio, 4.20 [95% CI, 1.61 to 6.29]). Initial Spo2 less than 98% (odds ratio, 2.76 [95% CI, 1.69 to 4.48]), extremes of weight (odds ratio, 2.18 [95% CI, 1.29 to 3.61]), and right-sided cases (odds ratio, 2.33 [95% CI, 1.41 to 3.92]) were associated with an increased risk of hypoxemia in the older cohort. Increasing age (1-yr increment; odds ratio, 0.88 [95% CI, 0.80 to 0.97]) was associated with a decreased risk of hypoxemia. An initial room air oxygen saturation of less than 98% was associated with an increased risk of hypoxemia in all children 4 to 17 yr of age. Extremes of weight, right-sided cases, and decreasing age were associated with an increased risk of hypoxemia in children 10 to 17 yr of age.


Biological resurfacing in a canine model of hip osteoarthritis.
Articular cartilage has unique load-bearing properties but has minimal capacity for intrinsic repair. Here, we used three-dimensional weaving, additive manufacturing, and autologous mesenchymal stem cells to create a tissue-engineered, bicomponent implant to restore hip function in a canine hip osteoarthritis model. This resorbable implant was specifically designed to function mechanically from the time of repair and to biologically integrate with native tissues for long-term restoration. A massive osteochondral lesion was created in the hip of skeletally mature hounds and repaired with the implant or left empty (control). Longitudinal outcome measures over 6 months demonstrated that the implant dogs returned to normal preoperative values of pain and function. Anatomical structure and functional biomechanical properties were also restored in the implanted dogs. Control animals never returned to normal and exhibited structurally deficient repair. This study provides clinically relevant evidence that the bicomponent implant may be a potential therapy for moderate hip osteoarthritis.


Making predictions in a changing world-inference, uncertainty, and learning.
To function effectively, brains need to make predictions about their environment based on past experience, i.e., they need to learn about their environment. The algorithms by which learning occurs are of interest to neuroscientists, both in their own right (because they exist in the brain) and as a tool to model participants' incomplete knowledge of task parameters and hence, to better understand their behavior. This review focusses on a particular challenge for learning algorithms-how to match the rate at which they learn to the rate of change in the environment, so that they use as much observed data as possible whilst disregarding irrelevant, old observations. To do this algorithms must evaluate whether the environment is changing. We discuss the concepts of likelihood, priors and transition functions, and how these relate to change detection. We review expected and estimation uncertainty, and how these relate to change detection and learning rate. Finally, we consider the neural correlates of uncertainty and learning. We argue that the neural correlates of uncertainty bear a resemblance to neural systems that are active when agents actively explore their environments, suggesting that the mechanisms by which the rate of learning is set may be subject to top down control (in circumstances when agents actively seek new information) as well as bottom up control (by observations that imply change in the environment).


Leukemia inhibitory factor production by rat mast cells.
Leukemia inhibitory factor (LIF) is a pluripotent cytokine of importance in the regulation of immune and inflammatory responses. This cytokine may play an important role in neuronal development and bone metabolism. We have examined the ability of freshly isolated rat mast cells and mast cell lines to produce LIF at both the mRNA and bioactivity levels. Initial experiments demonstrated that two mucosal mast cell-like cell lines RBL.2H3 and RCMC9 endogenously produced low levels of LIF bioactivity. The production of this cytokine was examined using a hepatocyte-stimulating factor activity assay and confirmed by the use of neutralizing antibodies specific for LIF. This production was enhanced by treatment with the calcium ionophore A23187. No interleukin-6 production was observed by these cells either endogenously or following ionophore activation. Freshly isolated highly purified rat peritoneal mast cells also expressed mRNA for LIF. These results could have important implications for the role of mast cells in neuronal development, hematopoiesis bone metabolism and the acute-phase response.


Quorum-sensing regulation governs bacterial adhesion, biofilm development, and host colonization in Pantoea stewartii subspecies stewartii.
The phytopathogenic bacterium Pantoea stewartii subsp. stewartii synthesizes stewartan exo/capsular polysaccharide (EPS) in a cell density-dependent manner governed by the EsaI/EsaR quorum-sensing (QS) system. This study analyzes biofilm development and host colonization of the WT and QS regulatory mutant strains of P. stewartii. First, we show that the cell density-dependent synthesis of stewartan EPS, governed by the EsaI/EsaR QS system, is required for proper bacterial adhesion and development of spatially defined, 3D biofilms. Second, a nonvirulent mutant lacking the esaI gene adheres strongly to surfaces and develops densely packed, less structurally defined biofilms in vitro. This strain appears to be arrested in a low cell density developmental mode. Exposure of this strain to exogenous N-acyl-homoserine lactone counteracts this adhesion phenotype. Third, QS mutants lacking the EsaR repressor attach poorly to surfaces and form amorphous biofilms heavily enmeshed in excess EPS. Fourth, the WT strain disseminates efficiently within the xylem, primarily in a basipetal direction. In contrast, the two QS mutant strains remain largely localized at the site of infection. Fifth, and most significantly, epifluorescence microscopic imaging of infected leaf tissue and excised xylem vessels reveals that the bacteria colonize the xylem with unexpected specificity, particularly toward the annular rings and spiral secondary wall thickenings of protoxylem, as opposed to indiscriminate growth to fill the xylem lumen. These observations are significant to bacterial plant pathogenesis in general and may reveal targets for disease control.


Unilateral destructive wrist synovitis in juvenile idiopathic arthritis.
To describe the clinical and radiographic features of a group of juvenile idiopathic arthritis (JIA) patients who developed unilateral destructive wrist synovitis. All wrist radiographs performed yearly between 1986 and 2002 in JIA patients who had wrist involvement were retrospectively reviewed to identify patients who had unilateral erosive wrist synovitis, defined as a difference of at least -3 units in the Poznanski score between the affected wrist and the unaffected wrist, with the Poznanski score in the unaffected wrist being > -2 units throughout the follow-up period. Clinical and radiographic data obtained during follow-up were recorded for all patients. Of a total of 250 patients for whom we had approximately 900 wrist radiographs, 6 patients were found to have unilateral erosive wrist synovitis. The JIA onset subtype was oligoarticular in 5 patients and polyarticular in 1 patient and the disease duration from presentation to the last follow-up visit ranged from 2 to 16 years. The arthritis course was polyarticular in all patients. Five patients had positive antinuclear antibodies (ANA) and 1 had positive rheumatoid factor (RF). At the last follow-up visit, all patients had some impairment of wrist function and 2 patients had wrist subluxation. There was a marked radiographic damage in all affected wrist, with the Poznanski ranging from -8.0 to -8.50 units in 3 patients and being -5.5, -3.1 and -2.4 units, respectively, in 3 patients. The severity of radiographic damage in the ANA-positive patients with the longest disease duration was comparable to that observed in the RF-positive patient. Unilateral erosive wrist synovitis seems to be uncommon in JIA. Patients with unilateral wrist synovitis may be at risk of a destructive course irrespective of the JIA onset subtype.


Berries as a Treatment for Obesity-Induced Inflammation: Evidence from Preclinical Models.
Inflammation that accompanies obesity is associated with the infiltration of metabolically active tissues by inflammatory immune cells. This propagates a chronic low-grade inflammation associated with increased signaling of common inflammatory pathways such as NF-κB and Toll-like receptor 4 (TLR4). Obesity-associated inflammation is linked to an increased risk of chronic diseases, including type 2 diabetes, cardiovascular disease, and cancer. Preclinical rodent and cell culture studies provide robust evidence that berries and their bioactive components have beneficial effects not only on inflammation, but also on biomarkers of many of these chronic diseases. Berries contain an abundance of bioactive compounds that have been shown to inhibit inflammation and to reduce reactive oxygen species. Therefore, berries represent an intriguing possibility for the treatment of obesity-induced inflammation and associated comorbidities. This review summarizes the anti-inflammatory properties of blackberries, blueberries, strawberries, and raspberries. This review highlights the anti-inflammatory mechanisms of berries and their bioactive components that have been elucidated through the use of preclinical models. The primary mechanisms mediating the anti-inflammatory effects of berries include a reduction in NF-κB signaling that may be secondary to reduced oxidative stress, a down-regulation of TLR4 signaling, and an increase in Nrf2.


Autonomic correlates of ocular accommodation and cardiovascular function.
To evaluate the hypothesis that objective measures of open- and closed-loop ocular accommodation are related to systemic cardiovascular function, and ipso facto autonomic nervous system activity. Sixty subjects (29 male; 31 female) varying in age from 18 to 33 years (average: 20.3 +/- 2.9 years) with a range of refractive errors [mean spherical equivalent (MSE): -7.12 to +1.82 D] participated in the study. Five 20-s continuous objective recordings of the accommodative response, measured with an open-view IR autorefractor (Shin-Nippon SRW-5000), were obtained for a variety of open- and closed-loop accommodative demands while simultaneous continuous measurement of heart rate was recorded with a finger-mounted piezo-electric pulse transducer for 5 min. Fast Fourier Transformation of cardiovascular function allowed the absolute and relative power of the autonomic components to be assessed in the frequency-domain, whereas heart period gave an indication of the time-domain response. Increasing closed-loop accommodative demand led to a concurrent increase in heart rate of approximately 2 beats/min for a 4.0 D increase in accommodative demand. The increase was attributable to a reduction in the absolute (p < 0.05) and normalised (p < 0.001) input of the systemic parasympathetic nervous system, and was unaffected by refractive group. The interaction with refractive group failed to reach significance. For sustained accommodation effort, the data demonstrate covariation between the oculomotor and cardiovascular systems which implies that a near visual task can significantly influence cardiovascular behaviour. Accommodative effort alone, however, is not a sufficient stimulus to induce autonomic differences between refractive groups. The data suggest that both the oculomotor and cardiovascular systems are predominantly attributable to changes in the systemic parasympathetic nervous system.


Application of multiblock analysis to identify key areas and risk factors for dairy cow persistence.
The present study analysed the importance of individual variables and different thematic blocks of production areas, management, and herd infectious disease status on cow persistence, characterised by herd on-farm mortality rate (MR), culling rate (CR), and mean age of culled cows (MAofCC) applying multiblock partial least squares (mbPLS) analysis. This study included 120 free-stall dairy herds with ≥ 100 cows. Data on the previous year's predominant cow housing system and management practices were collected, and on-farm measurements and cow scoring were performed. Bulk tank milk (BTM) and heifer blood samples (10 samples per herd) were collected and analysed for antibodies against the selected pathogens. In total, 172 variables were aggregated into 14 thematic blocks. The annual CR, MR, and MAofCC values were calculated for each herd. Thematic blocks with significant impact on cow persistence (included herd MR, CR and MAofCC) were 'infectious diseases' (block importance index out of all blocks = 13.6%, 95% CI 10.3; 20.5), 'fertility management' (16.3%, 95% CI 6.8; 26.9), 'lactating cow management' (11.5%, 95% CI 6.4; 17.8), 'milking' (11.3%, 95% CI 3.2; 17.1), 'herd characteristics' (10.1%, 95% CI 6.3; 14.2), 'close-up period management' (9.7%, 95% CI 2.7; 15.7), 'calving management' (7.9%, 95% CI 3.1; 11.4) and 'disease management' (7.3%, 95% CI 0.2; 12.0). Variable categories with the highest importance in explaining composite outcome including herd MR, CR and MAofCC were rear-end and udder lesions in ≥ 20% of the cows, BTM and heifers seropositive to bovine respiratory syncytial virus, vaccination against bovine herpesvirus 1, twice daily milking and herd location in Northwest region. Larger herd size, higher levels of milk yield, and rearing predominantly Holstein breed cattle were herd factors associated with poorer cow persistency. Grazing cows and having semi-insulated barns were associated with lower CR and MR, respectively. Heat detection and farm pregnancy testing strategies were significant factors in the fertility block. Using disposable dry papers for teat cleaning and not using any wet teat-cleaning tools were risk factors for high MR. A robotic milking system was protective for increased herd MR and CR. A high pre-calving body condition score and poor rear body cleanliness of ≥ 30% of cows were associated with inferior herd persistency outcomes. Calving in group pens with deep litter bedding was associated with a lower CR. Multiblock PLS model is innovative tool that helped to identify most influential farming areas but also single risk factors associated with cow persistency described by multiple parameters.


Increased production of colicin E1 in stationary phase.
The synthesis of colicin E1 is known to be regulated by the SOS response, anaerobiosis, and catabolite repression. The expression of cea-lacZ fusions was also found to be stimulated when cells reached stationary phase. This increase in expression was determined to be due to depletion of nutrients from the medium, since the addition of fresh medium reversed the effect. Expression of the fusion increased when cells were starved in 10 mM MgSO4 and when they were grown in conditioned medium in which cells had been grown previously. The stimulation of expression occurred when the cea-lacZ fusion was present in single-copy as well as in multicopy plasmids. Finally, the data were consistent with this increase being independent of the SOS response, anaerobiosis, catabolite repression, and integration host factor as well as the stationary-phase regulators encoded by rpoS and lrp.


Contact Location and Neuropsychological Outcomes in Subthalamic Deep Brain Stimulation.
A host of influences contribute to cognitive and behavioral changes following deep brain stimulation. The location of the active cathode is likely an important variable but it has received little attention. To determine whether active contact location relative to the subthalamic nucleus and other neighboring structures is related to nonmotor outcomes. We identified a retrospective, cross-sectional sample of 46 patients who underwent subthalamic nucleus deep brain stimulation for treatment of idiopathic Parkinson's disease. T-tests or nonparametric equivalents were used to detect baseline differences between unilateral left, unilateral right, and bilateral surgical groups. Correlation and partial correlational analyses identified relationships between contact location variables and alterations in cognitive, mood, quality of life, motor, and disease variables. Medial contact locations within the left subthalamic nucleus were correlated with improvements in self-reported mood (r12 = -0.78, P = .001; 95% confidence interval [CI] = -0.43 to -0.93) but worsening semantic fluency (r26 = -0.38, P = .048; 95% CI = -0.01 to -0.66). Phonemic fluency worsened with more posterior left placement (r34 = 0.35, P = .036; 95% CI = 0.03 to 0.61). Memory outcome was related to right hemisphere stimulation voltage (r29 = -0.40, P = .022; 95% CI = -0.05 to -0.66), which is likely a proxy for variable electrode location. Location of the active contact is related to nonmotor outcomes, even in electrodes that are adequately placed. This is relevant to clinical care as there appears to be a trade-off between mood and fluency abilities that should be considered during surgical planning according to preoperative patient characteristics.


Week 48 Resistance Analyses of the Once-Daily, Single-Tablet Regimen Darunavir/Cobicistat/Emtricitabine/Tenofovir Alafenamide (D/C/F/TAF) in Adults Living with HIV-1 from the Phase III Randomized AMBER and EMERALD Trials.
Darunavir/cobicistat/emtricitabine/tenofovir alafenamide (D/C/F/TAF) 800/150/200/10 mg is being investigated in two Phase III trials, AMBER (NCT02431247; treatment-naive adults) and EMERALD (NCT02269917; treatment-experienced, virologically suppressed adults). Week 48 AMBER and EMERALD resistance analyses are presented. Postbaseline samples for genotyping/phenotyping were analyzed from protocol-defined virologic failures (PDVFs) with viral load (VL) ≥400 copies/mL at failure/later time points. Post hoc analyses were deep sequencing in AMBER, and HIV-1 proviral DNA from baseline samples (VL <50 copies/mL) in EMERALD. Through week 48 across both studies, no darunavir, primary PI, or tenofovir resistance-associated mutations (RAMs) were observed in HIV-1 viruses of 1,125 participants receiving D/C/F/TAF or 629 receiving boosted darunavir plus emtricitabine/tenofovir-disoproxil-fumarate. In AMBER, the nucleos(t)ide analog reverse transcriptase inhibitor (N(t)RTI) RAM M184I/V was identified in HIV-1 of one participant during D/C/F/TAF treatment. M184V was detected pretreatment as a minority variant (9%). In EMERALD, in participants with prior VF and genoarchive data (N = 140; 98 D/C/F/TAF and 42 control), 4% had viruses with darunavir RAMs, 38% with emtricitabine RAMs, mainly at position 184 (41% not fully susceptible to emtricitabine), 4% with tenofovir RAMs, and 21% ≥ 3 thymidine analog-associated mutations (24% not fully susceptible to tenofovir) detected at screening. All achieved VL <50 copies/mL at week 48 or prior discontinuation. D/C/F/TAF has a high genetic barrier to resistance; no darunavir, primary PI, or tenofovir RAMs were observed through 48 weeks in AMBER and EMERALD. Only one postbaseline M184I/V RAM was observed in HIV-1 of an AMBER participant. In EMERALD, baseline archived RAMs to darunavir, emtricitabine, and tenofovir in participants with prior VF did not preclude virologic response.


Effect of the density of pillar-patterned substrates on contact mechanics: Transition from top to mixed contact with a detailed pressure-field description.
Different contact regimes between a spherical lens and a periodically patterned substrate are observed, when they are pressed against each other. Top contact occurs when only the highest substrate sections touch the lens, whereas mixed contact implies that both the highest and the lowest substrate sections come into contact with the lens. In this paper, we study how the pattern density of the substrate, along with its physical properties and those of the lens, determine the transition from top contact to mixed contact. Experiments and numerical simulations had been performed, as complementary approaches to obtain data, and a theoretical analysis has been developed to gain insight on the effect of the physical parameters on the contact transition. As a result, a phase diagram is presented, in terms of the load and the contact radius, that combines the observations of the three approaches (experimental, numerical, and theoretical), unveiling the boundaries of three contact regimes: (1) deterministic-driven contact, (2) top contact, and (3) mixed contact.


Metabolic studies with 5-fluorocytosine-6-14C in mouse, rat, rabbit, dog and man.
Metabolism of 5-fluorocytosine-6-14C (5-FC) was studied in mice, rats, rabbits and dogs after oral and subcutaneous, single and repeated administration. In the urines of all species, intact 5-FC accounted for more than 90% of the total radioactivity at any time of the various treatment schedules. The average proportion of the urinary metabolites was around 5% in dogs, 3% in rabbits, 2.5% in rats, and 2% in mice of the total radioactivity. At repeated dosage, there was an increase of metabolites in mice but a decrease in rats treated subcutaneously. Neither increase nor decrease was observed in rabbits (treated orally) and dogs. Two metabolites were identified, alpha-fluoro-beta-ureido-propionic acid (FUPA) and alpha-fluoro-beta-alanine, the latter occurring mainly after oral treatment. These compounds represent probably that part of 5-FC which was deaminated to 5-fluorouracil (5-FU) or directly to 5-fluorodihydrouracil. FUPA was the only metabolite found in the urines collected from 4 out of 5 human volunteers during the first 12 h after single oral administration of 3.5 g of the radiolabelled drug. Its maximum proportion was 1.1% of the total radioactivity. No metabolites were detected in the urine neither of the 5th volunteer nor in those of 3 mycosis patients who were given the radioactive dose after they had received regular chemotherapy with unlabelled 5-FC (150 mg/kg/day) for at least 2 weeks. The sensitivity threshold of the method was 0.1-0.4% of the total radioactivity. One of the patients had developed thrombocytopenia which was probably due to 5-FC chemotherapy. The symptoms of 5-FC intolerance were in most of the examined species similar to those observed with 5-FU [9]. However, no quantitative correlation between proportion of metabolites and 5-FC toxicity is apparent except that man is the species in which both metabolism and toxicity are the lowest. It has not been proved yet that 5-FC intolerance occurring in a small percentage of patients receiving 5-FC chemotherapy (mainly leukopenia, thrombocytopenia) results in fact from conversion to 5-FU.


Studies on mercury occurrence in inorganic constituents of Polish coking coals.
During the cokemaking process, a significant amount of mercury occurring in a coal blend is released to the atmosphere. One of the ways of reducing this emission is to reduce mercury content in a coal blend. This could be obtained through the coal washing process. The optimization of this process requires the knowledge of mercury occurrence in coal, especially in its inorganic constituents. A qualitative analysis of mercury occurrence in the inorganic constituents of Polish coking coals was performed using an electron probe microanalyzer (EPMA). For that purpose, selected samples of rejects and middling products derived from the washing process in dense media separators and jig concentrators were examined. The obtained results have confirmed a strong connection between mercury occurrence and the presence of sulfides (pyrite, marcasite, and chalcopyrite) in Polish coking coals. Significant amounts of mercury were also noticed for barite, siderite, and aluminosilicates. The highest value of mercury content, at the level of 0.100%, was obtained for marcasite. For the analyzed coals, the effectiveness of mercury removal in the washing process was determined by the forms of pyrite occurring in coal. The highest values of effectiveness of mercury removal were obtained in the case of coals for which the large framboidal pyrite aggregates with chalcopyrite overgrowths were noticed. It was also found that middling products were characterized by the occurrence of the Hg-rich overgrowths of pyrite on organic matter. To achieve a significant reduction in mercury content in clean coal, it is necessary to develop an effective method of removing this form of pyrite from hard coal.


[Effect of chitooligosaccharide on hepatic triglyceride metabolism and related mechanisms].
To investigate the effect of chitooligosaccharide (COS) on hepatic triglyceride (TG) metabolism and related mechanisms. The LO2 cells treated by 1 mmol/L fatty acid were used as model group, the cells treated by 1 mmol/L fatty acid and 0.5 mg/ml COS were used as COS group, and the untreated cells were used as control group. The TG content in cells was measured. RT-PCR and Western blot were used to measure the mRNA and protein expression of sterol regulatory element binding protein-1c (SREBP-1c), fatty acid synthase (FAS), and carnitine palmityl transferase 1A (CPT1A) in each group. Male C57BL/6J mice were randomized into control group, high-fat group, and COS group to receive different treatments. Sixteen weeks later, the liver was harvested for HE and oil red O staining to measure the content of TG in the liver. The t-test or one-way analysis of variance was used for comparison of data between groups, and the SNK method was used for comparison of data between any two groups. The LO2 cells in the model group had an increased number of lipid droplets and an increased TG content, and after COS treatment, the TG content was low. The COS group had significantly lower relative mRNA expression of SREBP-1c and FAS compared with the model group (1.135 ± 0.177 vs 2.322 ± 0.198,F= 60.457,P< 0.01; 1.226 ± 0.150 vs 1.801 ± 0.159,F= 24.753,P< 0.01), while compared with the control group, the COS group had significantly higher mRNA expression of CPT1A (1.254 ± 0.156 vs 1.908 ± 0.087,F= 31.734,P< 0.01). The COS group had significantly lower protein expression of SREBP-1c and FAS than the model group (0.161 ± 0.081 vs 0.351±0.016,F= 188.920,P< 0.01; 0.332 ± 0.023 vs 1.238 ± 0.051,F= 624.069,P< 0.01), and significantly higher protein expression of CPT1A than the model group (1.014 ± 0.033 vs 0.561 ± 0.046,F= 193.793,P< 0.01). COS reduced the TG content in the liver in rats on high-fat diet. COS can reduce the accumulation of TG in the hepatocyte model of nonalcoholic fatty liver disease and in the liver in rats on high-fat diet, and the possible mechanism may be related to inhibiting the expression of SREBP-1c and downstream FAS, reducing the synthesis of TG, increasing the expression of CPT1A, and accelerating the breakdown of TG.


Chronic Adolescent Δ9-Tetrahydrocannabinol Treatment of Male Mice Leads to Long-Term Cognitive and Behavioral Dysfunction, Which Are Prevented by Concurrent Cannabidiol Treatment.
Introduction: The high prevalence of adolescent cannabis use, the association between this use and later psychiatric disease, and increased access to high-potency cannabis highlight the need for a better understanding of the long-term effects of adolescent cannabis use on cognitive and behavioral outcomes. Furthermore, increasing Δ9-tetrahydrocannabinol (THC) in high-potency cannabis is accompanied by a decrease in cannabidiol (CBD), thus an understanding of the interactions between CBD and THC in the neurodevelopmental effects of THC is also important. The current study examined the immediate and long-term behavioral consequences of THC, CBD, and their combination in a mouse model of adolescent cannabis use. Materials and Methods: Male CD1 mice received daily injections of THC (3 mg/kg), CBD (3 mg/kg), CBD+THC (3 mg/kg each), vehicle, or remained undisturbed in their home cage (no handling/injections), either during adolescence (postnatal day [PND] 28-48) or during early adulthood (PND 69-89). Animals were then evaluated with a battery of behavioral tests 1 day after drug treatment, and again after 42 drug-free days. The tests included the following: open field (day 1), novel object recognition (NOR; day 2), marble burying (day 3), elevated plus maze (EPM; day 4), and Nestlet shredding (day 5). Results: Chronic administration of THC during adolescence led to immediate and long-term impairments in object recognition/working memory, as measured by the NOR task. In contrast, adult administration of THC caused immediate, but not long term, impairment of object/working memory. Adolescent chronic exposure to THC increased repetitive and compulsive-like behaviors, as measured by the Nestlet shredding task. Chronic administration of THC, either during adolescence or during adulthood, led to a delayed increase in anxiety as measured by the EPM. All THC-induced behavioral abnormalities were prevented by the coadministration of CBD+THC, whereas CBD alone did not influence behavioral outcomes. Conclusion: These data suggest that chronic exposure to THC during adolescence leads to some of the behavioral abnormalities common in schizophrenia. Interestingly, CBD appeared to antagonize all THC-induced behavioral abnormalities. These findings support the hypothesis that adolescent THC use can impart long-term behavioral deficits; however, cotreatment with CBD prevents these deficits.


Plasticity of intragraft alloreactive T cell clones in human gut correlates with transplant outcomes.
The site of transition between tissue-resident memory (TRM) and circulating phenotypes of T cells is unknown. We integrated clonotype, alloreactivity, and gene expression profiles of graft-repopulating recipient T cells in the intestinal mucosa at the single-cell level after human intestinal transplantation. Host-versus-graft (HvG)-reactive T cells were mainly distributed to TRM, effector T (Teff)/TRM, and T follicular helper compartments. RNA velocity analysis demonstrated a trajectory from TRM to Teff/TRM clusters in association with rejection. By integrating pre- and post-transplantation (Tx) mixed lymphocyte reaction-determined alloreactive repertoires, we observed that pre-existing HvG-reactive T cells that demonstrated tolerance in the circulation were dominated by TRM profiles in quiescent allografts. Putative de novo HvG-reactive clones showed a transcriptional profile skewed to cytotoxic effectors in rejecting grafts. Inferred protein regulon network analysis revealed upstream regulators that accounted for the effector and tolerant T cell states. We demonstrate Teff/TRM interchangeability for individual T cell clones with known (allo)recognition in the human gut, providing novel insight into TRM biology.


Current position of advanced laparoscopic surgery of the liver.
With the improvement of laparoscopic techniques and the development of new and dedicated technologies, endoscopic liver surgery has become feasible. While wedge liver resections are performed more and more frequently, laparoscopic anatomical liver resections are still at an early stage of development and are somewhat controversial. In 1993 we initiated formal laparoscopic liver resections in selected patients. From 1993 to December 1995 20 patients underwent endoscopic formal resections: the procedures comprised six left hepatectomies, five right hepatectomies, one of which extended to the segment IV, three mesohepatectomy, five segmentectomies and one bisegmentectomy. The operation time ranged from 120 to 270 min (average 193 min). In 17 out of 20 cases a Pringle manoeuvre was performed (mean occlusion time 45 min). No intra-operative complications occurred and there were no conversions in the whole series. Average intra-operative blood loss was 397.5 mL and 35% of patients required intro-operative blood transfusions. Post-operative mortality rate was 5% and post-operative morbidity rate was 45% (one coagulopathy with severe trombocytopaenia, six pleural effusions, one bile collection and four hematomas of the trocar sites). Such preliminary data are comparable with those of a group of 65 patients who underwent open anatomical liver resections from 1992 and 1995. Far from being a routine technique in liver surgery, the laparoscopic approach to forma liver resections may be a promising procedure in selected patients.


Neuropharmacological Aspects of Crocus sativus L.: A Review of Preclinical Studies and Ongoing Clinical Research.
Crocus sativus L. (Iridaceae) is an important member of the genus Crocus having high medicinal value. Its dried stigmas, known as "saffron" are being widely used form past many centuries as a food additive, coloring agent, flavoring agent and a potential source of traditional medicine. The stigmas along with other botanical parts of Crocus sativus are being extensively used in ethnomedical treatment of varied central nervous system diseases. In line with its ethnomedical importance, several preclinical studies have been carried out to validate its traditional uses, identify active principle(s), understand pharmacological basis of therapeutic action and explore novel medicinal uses. The bioactive components of Crocus sativus have been found to modulate several synaptic processes via direct/indirect interplay with neurotransmitter receptor functions, interaction with neuronal death/survival pathways and alteration in neuronal proteins expression. Many clinical studies proving beneficial effect of Crocus sativus in depressive disorders, Alzheimer's disease and some other neurological abnormalities have also been carried out. Based on the vast literature reports available, an attempt has been made to comprehend the fragmented information on neuropharmacological aspects, chemistry and safety of Crocus sativus. Although the plant has been well explored, but still a large scope of future preclinical and clinical research exist to explore its potential in neurological diseases, that has been discussed in depth in the present review.


Canine visceral leishmaniasis in the Northeast Region of Brazil.
Visceral leishmaniasis (VL) is a zoonosis that affects dogs and other mammals, including humans. Contact with dogs is a major risk factor for humans. This disease is endemic in several regions of Brazil. The aim of this study was to determine the prevalence of Leishmania spp. infection in dogs and to correlate it with possible risk factors. Blood samples were collected from 391 dogs of different ages, breeds, and both genders, coming from Campina Grande, Paraíba state, Brazil. An epidemiological questionnaire was employed in order to identify risk factors associated with the disease. Serological tests were performed using indirect immunofluorescence, enzyme-linked immunosorbent assay (ELISA S7®) and polymerase chain reaction. Leishmania spp. antibodies were detected in 33 (8.4 %) and 17 (4.3 %) dogs according to the indirect immunofluorescence test (IFAT) and enzyme-linked immunosorbent assay (ELISA S7®), respectively. PCR results indicated the presence of L. chagasi DNA in only eight (2 %) blood samples. There was a significant association between reactive animals and contact with animals from different houses (OR = 4.1; p = 0.02). It is suggested that CVL may occur in urban areas. Moreover, it is demonstrated that the association among different diagnostic tests may lead to a more accurate identification of positive animals, which might help to improve the disease control and prevent euthanasia in false-positive results.


Influence of physical restraint on unplanned extubation of adult intensive care patients: a case-control study.
Unplanned extubation commonly occurs in intensive care units. Various physical restraints have been used to prevent patients from removing their endotracheal tubes. However, physical restraint not only does not consistently prevent injury but also may be a safety hazard to patients. To evaluate the effect of physical restraint on unplanned extubation in adult intensive care patients. A total of 100 patients with unplanned extubations and 200 age-, sex-, and diagnosis-matched controls with no record of unplanned extubation were included in this case-control study. The 300 participants were selected from a population of 1455 patients receiving mechanical ventilation during a 21-month period in an adult intensive care unit at a medical center in Taiwan. Data were collected by reviewing medical records and incident reports of unplanned extubation. The incidence rate of unplanned extubation was 8.7%. Factors associated with increased risk for unplanned extubation included use of physical restraints (increased risk, 3.11 times), nosocomial infection (increased risk, 2.02 times), and a score of 9 or greater on the Glasgow Coma Scale on admission to the unit (increased risk, 1.98 times). Episodes of unplanned extubation also were associated with longer stays in the unit. An impaired level of consciousness on admission to the intensive care unit and the presence of nosocomial infection intensify the risk for unplanned extubation, even when physical restraints are used. To minimize the risk of unplanned extubation, nurses must establish better standards for using restraints.


Structural dynamics of HIV-1 Rev and its complexes with RRE and 5S RNA.
The Rev protein of the human immunodeficiency virus type 1 (HIV-1) has been studied by time-resolved fluorescence spectroscopy. The single tryptophan residue of Rev, Trp45, located within the arginine-rich RNA-binding domain of the protein, was utilized as an intrinsic spectroscopic probe. In addition, five peptides spanning different lengths of the arginine-rich domain, each containing the tryptophan residue, and two C-terminal deletion mutants of Rev, Rev M9 delta 14 and Rev M11 delta 14, were examined. Rev M9 delta 14 lacks residues 68-112 whereas Rev M11 delta 14 is missing residues 92-112 of the C-terminus of Rev. The fluorescence decay of Trp45 in wild-type Rev was resolved into four discrete lifetime components, and decay-associated spectra (DAS) were obtained for each component. The fluorescence decays of all five peptides and Rev M9 delta 14 were resolved into three lifetime components. The fluorescence decay of Rev M11 delta 14 was resolved into four components similar to those found for wild-type Rev. These results indicate that the activation domain (residues 78-93), present in wild-type Rev and Rev M11 delta 14, induced a unique tryptophan environment, characterized by a short-lived, blue-shifted emission, attributed to higher order assembly of Rev. In addition, fluorescence anisotropy decay data obtained for wild-type Rev and the two C-terminal deletion mutants also indicate that the activation domain mediates self-association of Rev. Based on the anisotropy decay results for wild-type Rev, the distribution of oligomers is independent of salt concentration. The average fluorescence lifetime of Trp45 was reduced upon complexation of Rev with a 40-mer fragment of the Rev response element containing the minimal element for Rev binding (F8-RRE), and the emission was blue-shifted. In addition, the local rotation of the tryptophan side chain was blocked in the protein-RRE complex. These results indicate that Trp45 directly interacts with the RRE. Rev is also shown to bind to 5S RNA, resulting in very similar changes in the time-resolved tryptophan fluorescence to those observed upon complexation of Rev with F8-RRE.


The relationships of the Euparkeriidae and the rise of Archosauria.
For the first time, a phylogenetic analysis including all putative euparkeriid taxa is conducted, using a large data matrix analysed with maximum parsimony and Bayesian analysis. Using parsimony, the putative euparkeriid Dorosuchus neoetus from Russia is the sister taxon to Archosauria + Phytosauria. Euparkeria capensis is placed one node further from the crown, and forms a euparkeriid clade with the Chinese taxa Halazhaisuchus qiaoensis and 'Turfanosuchus shageduensis' and the Polish taxon Osmolskina czatkowicensis. Using Bayesian methods, Osmolskina and Halazhaisuchus are sister taxa within Euparkeriidae, in turn sister to 'Turfanosuchus shageduensis' and then Euparkeria capensis. Dorosuchus is placed in a polytomy with Euparkeriidae and Archosauria + Phytosauria. Although conclusions remain tentative owing to low node support and incompleteness, a broad phylogenetic position close to the base of Archosauria is confirmed for all putative euparkeriids, and the ancestor of Archosauria +Phytosauria is optimized as similar to euparkeriids in its morphology. Ecomorphological characters and traits are optimized onto the maximum parsimony strict consensus phylogeny presented using squared change parsimony. This optimization indicates that the ancestral archosaur was probably similar in many respects to euparkeriids, being relatively small, terrestrial, carnivorous and showing relatively cursorial limb morphology; this Bauplan may have underlain the exceptional radiaton and success of crown Archosauria.


Ontogeny of intestinal safety factors: lactase capacities and lactose loads.
We measured intestinal safety factors (ratio of a physiological capacity to the load on it) for lactose digestion in developing rat pups. Specifically, we assessed the quantitative relationships between lactose load and the series capacities of lactase and the Na+-glucose cotransporter (SGLT-1). Both capacities increased significantly with age in suckling pups as a result of increasing intestinal mass and maintenance of mass-specific activities. The youngest pups examined (5 days) had surprisingly high safety factors of 8-13 for both lactase and SGLT-1, possibly because milk contains lactase substrates other than lactose; it also, however, suggests that their intestinal capacities were being prepared to meet future demands rather than just current ones. By day 10 (and also at day 15), increased lactose loads resulted in lower safety factors of 4-6, values more typical of adult intestines. The safety factor of SGLT-1 in day 30 (weanling) and day 100 (adult) rats was only approximately 1.0. This was initially unexpected, because most adult intestines maintain a modest reserve capacity beyond nutrient load values, but postweaning rats appear to use hindgut fermentation, assessed by gut morphology and hydrogen production assays, as a built-in reserve capacity. The series capacities of lactase and SGLT-1 varied in concert with each other over ontogeny and as lactose load was manipulated by experimental variation in litter size.


The eIF4G-eIF4E complex is the target for direct cleavage by the rhinovirus 2A proteinase.
The 2A proteinases (2Apro) of certain picornaviruses induce the cleavage of the eIF4G subunit of the cap-binding protein complex, eIF4F. Several reports have demonstrated that 2Apro of rhinovirus and coxsackievirus B4 cleave eIF4G directly. However, it was suggested that in poliovirus infection, the 2Apro induces the activation of a cellular proteinase which in turn cleaves eIF4G. Furthermore, it is not clear whether eIF4G is cleaved as part of the eIF4F complex or as an individual polypeptide. To address these issues, recombinant eIF4G was purified from Sf9 insect cells and tested for cleavage by purified rhinovirus 2Apro. Here we report that eIF4G alone is a relatively poor substrate for cleavage by the rhinovirus 2Apro. However, an eIF4G-eIF4E complex is cleaved efficiently by the 2Apro, suggesting that eIF4F is a preferred substrate for cleavage by rhinovirus 2Apro. Furthermore, 2Apr drastically reduced the translation of a capped mRNA. An eIF4G-eIF4E complex, but not eIF4G alone, was required to restore translation.


Mutational analysis of a virulence locus in the E2 glycoprotein gene of Sindbis virus.
The substitution of arginine for serine at position 114 of glycoprotein E2 in several biological and recombinant Sindbis virus mutants was shown previously to attenuate the virus for neonatal mice and also to accelerate virus penetration into BHK cells. To further examine the genetically linked effects on both virus penetration into cultured cells and pathogenesis in vivo, mutants containing each of 16 different amino acid coding changes at this position were generated by site-directed mutagenesis of a full-length cDNA clone of the Sindbis virus genome. Viable virus was recovered following transfection of RNA transcripts from 14 of the clones. Phenotypic analysis of these virus mutants revealed that specific amino acid residues affected either the pathogenesis or penetration phenotype independently or both phenotypes simultaneously. Thus, both the position of a mutation within the E2 sequence and the particular amino acid encoded at that position are important determinants of the mutant phenotypes.


Production of the interferon protein: hyporesponsiveness.
A number of mechanisms may be responsible for development of hyporeactivity in cells or animals infected with viruses or exposed to interferon inducers. Available evidence, however, has implicated a specific control protein in modulation of interferon production. The precise identity and mechanism by which the repressor molecule modulates interferon production is at present unknown. Development of hyporeactivity to interferon induction as a consequence of repeated doses of an inducer or that develops secondary to virus infection may represent a serious limitation to use of inducers as antiviral agents. However, judicious selection of treatment regimens and inducers may circumvent or limit the severity of hyporeactivity and consequently enhance the therapeutic activity of interferon inducers.