The CDC14A phosphatase regulates oocyte maturation in mouse.
The final steps of oogenesis occur during oocyte maturation that generates fertilization-competent haploid eggs capable of supporting embryonic development. Cyclin-dependent kinase 1 (CDK1) drives oocyte maturation and its activity and actions on substrates are tightly regulated. CDC14 is a dual-specificity phosphatase that reduces CDK1 activity and reverses the actions of CDK1 during mitosis. In budding yeast, Cdc14 is essential for meiosis, but it is not known whether its mammalian homolog CDC14A is required for meiosis in females. Here, we report that CDC14A is concentrated in the nucleus of meiotically incompetent mouse oocytes but is dispersed throughout meiotically competent oocytes. During meiotic progression CDC14A has no specific sub-cellular localization except between metaphase of meiosis I (Met I) and metaphase of meiosis II (Met II) when it co-localizes with the central portion of the meiotic spindle. Overexpression of CDC14A generally delays meiotic progression after resumption of meiosis whereas microinjection of oocytes with an antibody against CDC14A specifically delays exit from Met I. Each of these perturbations generates eggs with chromosome alignment abnormalities and eggs that were injected with the CDC14A antibody had an elevated incidence of aneuploidy. Collectively, these data suggest that CDC14A regulates oocyte maturation and functions to promote the meiosis I-to-meiosis II transition as its homolog does in budding yeast.


System of systems engineering and risk management of extreme events: concepts and case study.
The domain of risk analysis is expanded to consider strategic interactions among multiple participants in the management of extreme risk in a system of systems. These risks are fraught with complexity, ambiguity, and uncertainty, which pose challenges in how participants perceive, understand, and manage risk of extreme events. In the case of extreme events affecting a system of systems, cause-and-effect relationships among initiating events and losses may be difficult to ascertain due to interactions of multiple systems and participants (complexity). Moreover, selection of threats, hazards, and consequences on which to focus may be unclear or contentious to participants within multiple interacting systems (ambiguity). Finally, all types of risk, by definition, involve potential losses due to uncertain events (uncertainty). Therefore, risk analysis of extreme events affecting a system of systems should address complex, ambiguous, and uncertain aspects of extreme risk. To accomplish this, a system of systems engineering methodology for risk analysis is proposed as a general approach to address extreme risk in a system of systems. Our contribution is an integrative and adaptive systems methodology to analyze risk such that strategic interactions among multiple participants are considered. A practical application of the system of systems engineering methodology is demonstrated in part by a case study of a maritime infrastructure system of systems interface, namely, the Straits of Malacca and Singapore.


Analysis of mtDNA sequence variants in colorectal adenomatous polyps.
Colorectal tumors mostly arise from sporadic adenomatous polyps. Polyps are defined as a mass of cells that protrudes into the lumen of the colon. Adenomatous polyps are benign neoplasms that, by definition display some characteristics of dysplasia. It has been shown that polyps were benign tumors which may undergo malignant transformation. Adenomatous polyps have been classified into three histologic types; tubular, tubulovillous, and villous with increasing malignant potential. The ability to differentially diagnose these colorectal adenomatous polyps is important for therapeutic intervention. To date, little efforts have been directed to identifying genetic changes involved in adenomatous polyps. This study was designed to examine the relevance of mitochondrial genome alterations in the three adenomatous polyps. Using high resolution restriction endonucleases and PCR-based sequencing, fifty-seven primary fresh frozen tissues of adenomatous polyps (37 tumors and 20 matched surrounding normal tissues) obtained from the southern regional Cooperative Human Tissue Network (CHTN) and Grady Memorial Hospital at Atlanta were screened with three mtDNA regional primer pairs that spanned 5.9 kbp. Results from our data analyses revealed the presence of forty-four variants in some of these mitochondrial genes that the primers spanned; COX I, II, III, ATP 6, 8, CYT b, ND 5, 6 and tRNAs. Based on the MITODAT database as a sequence reference, 25 of the 44 (57%) variants observed were unreported. Notably, a heteroplasmic variant C8515G/T in the MT-ATP 8 gene and a germline variant 8327delA in the tRNAlys was observed in all the tissue samples of the three adenomatous polyps in comparison to the referenced database sequence. A germline variant G9055A in the MT-ATP 6 gene had a frequency of 100% (17/17) in tubular and 57% (13/23) in villous adenomas; no corresponding variant was in tubulovillous adenomas. Furthermore, A9006G variant at MT-ATP 6 gene was observed at frequency of 57% (13/23) in villous adenomas only. Interestingly, variants A9006G and G9055A were absent in the villous tissue samples that were clinicopathological designated as "polyvillous adenomas". Our current data provide a basis for continued investigation of certain mtDNA variants as predictors of the three adenomatous polyps in a larger number of clinicopathological specimens.


On being fired: experiences of patient-initiated termination of the patient-physician relationship in palliative medicine.
Being "fired'' by a patient is an uncomfortable, distressing, and awkward experience for most clinicians. Palliative care clinicians may be at increased risk of termination of the patient-physician relationship for a variety of reasons. Little has been written about the experience of patient-initiated termination of the patient-physician relationship, and to our knowledge, nothing has been written that is specific to palliative care. To discuss the experience of termination, offer a framework for learning from and coping with the experience, and make recommendations for how to meet patients' needs after the termination. Case study and conceptual analysis of three cases from the experience of a palliative care fellow. The experience of patient-initiated termination of the patient-physician relationship can raise issues of rejection and self-doubt in the palliative care clinician, but can also provide important opportunities for growth, reflection, and learning. In some cases, re-connecting with the patients' families may be appropriate and beneficial for both the clinician and family.


The nursing workforce: a comparison of three national surveys.
The termination of the National Sample Survey of Registered Nurses (NSSRN) represents a loss of a key source of information on the nursing workforce that has been available for more than 30 years. At the same time, this loss presents new opportunities to address some of the biases associated with using licensing data to construct lists of RNs to construct a sampling frame. Given the loss of this key national data source on the nursing workforce, the NSSRN is compared to existing alternatives for nursing workforce data in two U.S. Census Bureaus surveys: the Current Population Survey and the American Communities Survey. The endeavor of workforce planning and a deeper understanding of whether we are equipped to meet the nation's changing health care landscape will benefit from a thoughtful consideration of how best to obtain accurate and comparable data from future surveys.


High-resolution echo-planar spectroscopic imaging of the human calf.
This study exploits the speed benefits of echo-planar spectroscopic imaging (EPSI) to acquire lipid spectra of skeletal muscle. The main purpose was to develop a high-resolution EPSI technique for clinical MR scanner, to visualise the bulk magnetic susceptibility (BMS) shifts of extra-myocellular lipid (EMCL) spectral lines, and to investigate the feasibility of this method for the assessment of intra-myocellular (IMCL) lipids. The study group consisted of six healthy volunteers. A two dimensional EPSI sequence with point-resolved spectroscopy (PRESS) spatial localization was implemented on a 3T clinical MR scanner. Measurements were performed by means of 64×64 spatial matrix and nominal voxel size 3×3×15 mm(3). The total net measurement time was 3 min 12 sec for non-water-suppressed (1 acquisition) and 12 min 48 sec for water-suppressed scans (4 acquisitions). Spectra of the human calf had a very good signal-to-noise ratio and linewidths sufficient to differentiate IMCL resonances from EMCL. The use of a large spatial matrix reduces inter-voxel signal contamination of the strong EMCL signals. Small voxels enabled visualisation of the methylene EMCL spectral line splitting and their BMS shifts up to 0.5 ppm relative to the correspondent IMCL line. The mean soleus muscle IMCL content of our six volunteers was 0.30±0.10 vol% (range 0.18-0.46) or 3.6±1.2 mmol/kg wet weight (range: 2.1-5.4). This study demonstrates that high-spatial resolution PRESS EPSI of the muscle lipids is feasible on standard clinical scanners.


Evaluating a common semi-mechanistic mathematical model of gene-regulatory networks.
Modeling and simulation of gene-regulatory networks (GRNs) has become an important aspect of modern systems biology investigations into mechanisms underlying gene regulation. A key challenge in this area is the automated inference (reverse-engineering) of dynamic, mechanistic GRN models from gene expression time-course data. Common mathematical formalisms for representing such models capture two aspects simultaneously within a single parameter: (1) Whether or not a gene is regulated, and if so, the type of regulator (activator or repressor), and (2) the strength of influence of the regulator (if any) on the target or effector gene. To accommodate both roles, "generous" boundaries or limits for possible values of this parameter are commonly allowed in the reverse-engineering process. This approach has several important drawbacks. First, in the absence of good guidelines, there is no consensus on what limits are reasonable. Second, because the limits may vary greatly among different reverse-engineering experiments, the concrete values obtained for the models may differ considerably, and thus it is difficult to compare models. Third, if high values are chosen as limits, the search space of the model inference process becomes very large, adding unnecessary computational load to the already complex reverse-engineering process. In this study, we demonstrate that restricting the limits to the [-1, +1] interval is sufficient to represent the essential features of GRN systems and offers a reduction of the search space without loss of quality in the resulting models. To show this, we have carried out reverse-engineering studies on data generated from artificial and experimentally determined from real GRN systems.


A multi-scale approach to designing therapeutics for tuberculosis.
Approximately one third of the world's population is infected with Mycobacterium tuberculosis. Limited information about how the immune system fights M. tuberculosis and what constitutes protection from the bacteria impact our ability to develop effective therapies for tuberculosis. We present an in vivo systems biology approach that integrates data from multiple model systems and over multiple length and time scales into a comprehensive multi-scale and multi-compartment view of the in vivo immune response to M. tuberculosis. We describe computational models that can be used to study (a) immunomodulation with the cytokines tumor necrosis factor and interleukin 10, (b) oral and inhaled antibiotics, and


Different responses of active and inactive plasma renin to various stimuli.
1. The response of active and inactive plasma renin to orthostasis and frusemide and to inhibition of prostaglandin synthesis by indomethacin was tested in normal human volunteers. 2. Active renin increased by orthostasis and frusemide and decreased by indomethacin. The latter also blunted the increase of active renin after stimuli. 3. Inactive renin was slightly increased by orthostasis, but was not significantly influenced by acute administration of frusemide or inhibition of prostaglandin synthesis by indomethacin. 4. The results show differences in the response of active and inactive renin to stimuli and suppression. Opposite changes of active and inactive renin were not observed in the experimental conditions studied.


Associations of dietary fat with albuminuria and kidney dysfunction.
Diet represents a potentially important target for intervention in nephropathy, yet data on this topic are scarce. The objective was to investigate associations between dietary fats and early kidney disease. We examined cross-sectional associations between dietary fats and the presence of high albuminuria (an established independent predictor of kidney function decline, cardiovascular disease, and mortality) or estimated glomerular filtration rate (eGFR) <60 mL ⋅ min(-1) ⋅ 1.73 m(-2) at baseline in 19,256 participants of the REGARDS (Reasons for Geographic and Racial Differences in Stroke) study, an ongoing cohort study in US adults aged ≥45 y at time of enrollment. We used logistic regression to assess associations between quintiles of total fat and subtypes of dietary fat (saturated, monounsaturated, polyunsaturated, and trans fat) and presence of high albuminuria or eGFR <60 mL ⋅ min(-1) ⋅ 1.73 m(-2). After multivariable adjustment, only saturated fat intake was significantly associated with high albuminuria [for quintile 5 compared with quintile 1, odds ratio (OR): 1.33; 95% CI: 1.07, 1.66; P for trend = 0.04]. No significant associations between any type of fat and eGFR <60 mL · min(-1) · 1.73 m(-2) were observed. ORs between the highest quintile of saturated fat and eGFR <60 mL · min(-1) · 1.73 m(-2) varied by race with a borderline significant interaction term (ORs: 1.24 in whites compared with 0.74 in blacks; P for interaction = 0.05) in multivariable-adjusted models, but no other associations were significantly modified by race or diabetes status. Higher saturated fat intake is significantly associated with the presence of high albuminuria, but neither total nor other subtypes of dietary fat are associated with high albuminuria or eGFR <60 mL · min(-1) · 1.73 m(-2).


Preoperative risk assessment for carotid occlusion by transcranial Doppler ultrasound.
An endovascular carotid balloon occlusion test with continuous intracranial monitoring by transcranial Doppler sonography was performed in 55 patients for prediction of tolerance of a required permanent occlusion of the carotid artery. Blood flow velocities of the ipsilateral middle cerebral artery during occlusion were recorded and compared with clinical tolerance during an occlusion test as well as with postoperative outcome after an eventual permanent occlusion. To stress the capacity of the cerebral circulation to tolerate the occlusion acetazolamide was injected before occlusion in all patients. The onset of neurological symptoms during temporary occlusion was dependent on the percentage fall of mean blood flow velocity relative to baseline rather than on absolute flow velocities during the time of occlusion. Patients with a fall of mean flow velocity of less than 30% tolerated temporary and permanent occlusion, with the exception of two patients who developed an infarction due to thromboembolism after iatrogenic sacrifice of the carotid artery. Patients with a major decrease developed neurological symptoms during occlusion in 55% and, in cases of carotid ligation, a haemodynamic infarction occurred. The results show that transcranial Doppler monitoring as a part of an endovascular balloon occlusion test may be a reliable technique for preoperative risk assessment for permanent occlusion of the carotid artery.


[Jinghuaweikang capsules combined with triple therapy in the treatment of Helicobacter pylori associated gastritis and duodenal ulcer and analysis of antibiotic resistance: a multicenter, randomized, controlled, clinical study].
To explore the efficacy of Jinghuaweikang capsules plus triple therapy (LACJ) in treatment of Helicobacter pylori (H. pylori) associated gastritis or duodenal ulcer, compare it with bismuth-containing quadruple therapy (LACB) and standard triple therapy (LAC) and analyze the antibiotic sensitivity of gastric mucosal H. pylori strains from the failed patients. A total of 565 patients with H. pylori infection were recruited from 11 hospitals from January 2010 to June 2011. There were 336 males and 229 females. They underwent gastroendoscopy examination due to upper gastrointestinal symptoms and had never received H. pylori eradication therapies. Duodenal ulcer patients were divided randomly into LACJ therapy group, LACB therapy group and LAC therapy group while gastritis patients LACJ therapy group and LACB therapy group. Group LAC received lansoprazole 30 mg + amoxicillin 1000 mg + clarithromycin 500 mg, twice a day, for 7 d (d1-7). Group LACJ: LAC therapy plus Jinghuaweikang, 3 capsules, twice a day, for 7 d (d1-7) then Jinghuaweikang, 3 capsules, twice a day, for 14 d (d8-21). Group LACB: LAC plus bismuth potassium citrate 220 mg, twice a day, for 7 d (d1-7) and then bismuth potassium citrate 220 mg, twice a day, for 14 d (d8-21). All duodenal ulcer patients received lansoprazole (30 mg, once a day) for 14 days after the first 7-day of treatment (d 8-21). At least 28 days after the end of treatment, all patients underwent (13)C urea breath test. Gastric mucosa was collected under endoscopy from the failed patients. The detection technique of gene chip was employed to detect antibiotics resistant gene from mucosa. The eradication rates of duodenal ulcer patients in groups LACJ, LACB and LAC were as follows: per-protocol (PP), 80.2% (77/96), 89.9% (89/99) and 72.2% (70/97) (P = 0.007), intention-to-treat (ITT), 78.6% (77/98), 88.1% (89/101) and 70.0% (70/100) (P = 0.007). No statistical differences existed between groups LACJ and LACB or LAC (all P > 0.05). But there were statistical differences between groups LACB and LAC (both P = 0.002). The eradication rates of PP and ITT of chronic gastritis patients in groups LACJ and LACB were as follows: 75.8% (97/128), 74.6% (97/130) vs 83.8% (109/130), 80.1% (109/136) (both P > 0.05). The symptomatic improvements of abdominal pain, burning and acid reflux of duodenal ulcer patients in group LACJ were higher than those in groups LACB and LAC. There were statistical differences between groups LACJ and LAC (all P < 0.05). The symptomatic improvements of bloating and belching for chronic gastritis patients in group LACJ were higher than those of group LACB. But no significant difference existed between two groups (all P > 0.05). Sixty samples of gastric mucosa were collected from the failed patients. The detection rates of antibiotic-resistant gene to clarithromycin and amoxicillin were 60.0% (36/36) and 18.3% (11/60) respectively. The efficacy of LACJ for the treatment of H. pylori infection patients is similar to LACB and superior to LAC. And the symptomatic improvement of patients is better than the other two regimens. The main cause of treatment failure is antibiotic resistance of H. pylori strains.


The Artificial Synapse Chip: a flexible retinal interface based on directed retinal cell growth and neurotransmitter stimulation.
The Artificial Synapse Chip is an evolving design for a flexible retinal interface that aims to improve visual resolution of an electronic retinal prosthesis by addressing cells individually and mimicking the physiological stimulation achieved in synaptic transmission. We describe three novel approaches employed in the development of the Artificial Synapse Chip: (i) micropatterned substrates to direct retinal cell neurite growth to individual stimulation sites; (ii) a prototype retinal interface based on localized neurotransmitter delivery; and (iii) the use of soft materials to fabricate these devices. By patterning the growth of cells to individual stimulation sites, we can improve the selectivity of stimulation and decrease the associated power requirements. Moreover, we have microfabricated a neurotransmitter delivery system based on a 5- micro m aperture in a 500-nm-thick silicon nitride membrane overlying a microfluidic channel. This device can release neurotransmitter volumes as small as 2 pL, demonstrating the possibility of chemical-based prostheses. Finally, we have fabricated and implanted an equivalent device using soft flexible materials that conform to the retinal tissue more effectively. As many of the current retinal prosthesis devices use hard materials and electrical excitation at a lower resolution, our approach may provide more physiologic retinal stimulation.


Malaria control in Papua New Guinea results in complex epidemiological changes.
With a renewed interest in large-scale malaria interventions, knowledge about the possible long-term effects of such interventions on the nature of malaria transmission is essential. We document complex changes in malaria epidemiology over the last 40 years associated with changing malaria control activities in Karimui, an isolated area in Papua New Guinea. An initially equal distribution of Plasmodium falciparum, P. vivax and P. malariae changed to currently 68% P. falciparum, after passing through a phase of transitory P. vivax dominance, when control started to fail. Initial drops in malaria prevalence proved difficult to sustain and present post-control levels are significantly higher than pre-control levels. The example of Karimui indicates that unsustained control can lead to changes in malaria patterns that may leave a population worse off.


Sudden sensorineural hearing loss is associated with chronic rhinosinusitis: population-based study.
This study aimed to evaluate the association of chronic rhinosinusitis with sudden sensorineural hearing loss using a population-based database. Sampled subject data were obtained from the Taiwan Longitudinal Health Insurance Database 2000. A total of 3325 patients with sudden sensorineural hearing loss were identified and 9975 controls were randomly selected. A conditional logistic regression was used to calculate the odds ratio for having been previously diagnosed with chronic rhinosinusitis, for cases and controls. The adjusted odds ratio of having prior chronic rhinosinusitis among cases compared to controls was 1.36 (95 per cent confidence interval = 1.16-1.60). The significant relationship between sudden sensorineural hearing loss and chronic rhinosinusitis was most pronounced among those patients aged 44 years or less (compared to controls) (odds ratio = 2.18; 95 per cent confidence interval = 1.63-2.92). However, the significant relationship between sudden sensorineural hearing loss and prior chronic rhinosinusitis was not sustained for patients older than 60 years compared to controls.


Energy-efficient coding with discrete stochastic events.
We investigate the energy efficiency of signaling mechanisms that transfer information by means of discrete stochastic events, such as the opening or closing of an ion channel. Using a simple model for the generation of graded electrical signals by sodium and potassium channels, we find optimum numbers of channels that maximize energy efficiency. The optima depend on several factors: the relative magnitudes of the signaling cost (current flow through channels), the fixed cost of maintaining the system, the reliability of the input, additional sources of noise, and the relative costs of upstream and downstream mechanisms. We also analyze how the statistics of input signals influence energy efficiency. We find that energy-efficient signal ensembles favor a bimodal distribution of channel activations and contain only a very small fraction of large inputs when energy is scarce. We conclude that when energy use is a significant constraint, trade-offs between information transfer and energy can strongly influence the number of signaling molecules and synapses used by neurons and the manner in which these mechanisms represent information.


Morphomechanical programming of morphogenesis in cnidarian embryos.
The factors governing the pattern formation process in the early morphogenesis of a marine colonial hydroid, Dynamena pumila, have been studied. Two different types of morphogenesis have been distinguished. Morphogenesis of the first type goes on via changes in cell shape and cell axis orientation, while morphogenesis of the second type is based upon the active coordinated cell movements associated with cell rearrangements. It was shown that morphogenesis of both types can be considered as cascades in which any event is a consequence of the previous one. The spatial structure of each developmental stage contains information about the direction and the initial conditions of further morphogenesis. So, an "epigenetic program" of morphogenesis gradually originates in the course of development and provides the stable reproduction of spatial structures. It is reasonable to consider the activity of epigenetic factors guiding Dynamena morphogenesis (geometry/topology of an embryo, heterogeneity of an embryo spatial structure, configuration of the field of mechanical stresses of the embryo surface) as "morphomechanical programming" of morphogenesis.


The effect of opioid receptor blockade on the neural processing of thermal stimuli.
The endogenous opioid system represents one of the principal systems in the modulation of pain. This has been demonstrated in studies of placebo analgesia and stress-induced analgesia, where anti-nociceptive activity triggered by pain itself or by cognitive states is blocked by opioid antagonists. The aim of this study was to characterize the effect of opioid receptor blockade on the physiological processing of painful thermal stimulation in the absence of cognitive manipulation. We therefore measured BOLD (blood oxygen level dependent) signal responses and intensity ratings to non-painful and painful thermal stimuli in a double-blind, cross-over design using the opioid receptor antagonist naloxone. On the behavioral level, we observed an increase in intensity ratings under naloxone due mainly to a difference in the non-painful stimuli. On the neural level, painful thermal stimulation was associated with a negative BOLD signal within the pregenual anterior cingulate cortex, and this deactivation was abolished by naloxone.


Tethered spinal cord syndrome in adults in the MRI era: recognition, pathology, and long-term objective outcomes.
Tethered cord syndrome (TCS) has been well described in pediatric patients. Many recent reports of TCS in adult patients have grouped retethering patients with newly diagnosed ones without separately analyzing each entity and outcome. The authors reviewed their experience of newly diagnosed adult TCS patients to identify and explore TCS misdiagnosis, recognition, subtype pathology, and individual objective outcomes. This study included 24 adult patients (20 female and 4 male) who fit the criteria of being newly diagnosed and aged 20 years and older (age range 20-77 years). Preexisting dermal sinus was present in 6 patients, hypertrichosis in 5, skin tag/cleft/dimple and fatty subcutaneous masses in 5, scoliosis in 2, and neurological abnormalities in 4 patients. The pathology consisted of TCS with taut filum in 8 patients, conus lipoma with TCS in 7, diastematomyelia in 7, and cervical cord tethering in 2 patients. Of the 24 study patients, nondermatomal low-back or perineal pain occurred in 19 patients, bladder dysfunction in 21, and motor, sensory, and reflex abnormalities in 21 patients. Aggravating factors were repeated stretching, multiple pregnancies, heavy lifting, and repeated bending. Urological evaluation included bladder capacity, emptying, postvoid residuals, detrusor function, pelvic floor electromyography (EMG), bladder sensitivity, and sphincter EMG, which were repeated at 6 months and 1 year postoperatively. The follow-up was 1 to 30 years. Detailed postoperative neurological findings and separate patient outcome evaluations were recorded. Four of the 24 patients did not have an operation. Resolution of pain occurred in 16 of the 19 patients reporting low-back or perineal pain. Motor and sensory complaints resolved in 17 of 20 patients. Regarding bladder dysfunction, in the 20 patients with available data, bladder function returned to normal in 12 patients, improved in 3 patients, and was unchanged in 5 patients. If the symptom duration was less than 6-8 months, there was recovery of all parameters of pain, bladder dysfunction, and neurological deficit, and recovery from hyperreflexia matched that from neurological deficit. Fifteen patients were employed preoperatively and returned to work, and an additional 3 others who were unable to work preoperatively were able to do so postoperatively. Most adults with newly diagnosed TCS have unrecognized neurocutaneous abnormalities and neurological deficits. The triad of nondermatomal sacral or perineal pain, bladder dysfunction, and neurological deficit should not be confused with hip or degenerative lumbosacral disease. Addressing the primary pathology often leads to successful results.


Ultrastructural analysis of the Fisher to Lewis rat model of chronic allograft nephropathy.
Chronic allograft nephropathy (CAN) is the leading cause of graft loss following kidney transplantation. One factor contributing to CAN is chronic alloimmune injury. However, the involvement of alloantigen-dependent and -independent factors in CAN is unclear. The pathomechanism of CAN has been extensively studied by utilizing the Fischer-to-Lewis (F344-to-LEW) rat model. Transplant capillaropathy (circumferential multiplication of the peritubular capillary basement membrane) and transplant glomerulopathy (reduplication of the glomerular basement membrane) have recently been validated clinicopathologically as ultrastructural indicators of chronic alloimmune injury. To investigate the presence of these markers, F344-to-LEW kidneys were examined by electron and light microscopy 32, 40 and 52 weeks after implantation. F344 rats with or without 30-min ischemia of the left kidney following right nephrectomy served as controls. All transplanted rats displayed marked proteinuria. On electron microscopy, transplant capillaropathy, transplant glomerulopathy, and T-cell cytotoxicity (indicator of ongoing cellular rejection) were absent. On light microscopy, the arteries were devoid of intimal fibrosis. Focal-segmental glomerulopathy resembling hyperfiltration injury was encountered, with mild interstitial infiltration, fibrosis, and tubular atrophy. The proteinuria and kidney pathology were more severe in transplanted than in ischemic or uninephrectomized rats. Because chronic-active rejection could not be detected between weeks 32 and 52, we propose that the alloantigen-dependent initial graft injury subsides, but induces the late events: glomerular hyperfiltration, proteinuria, and glomerulosclerosis. Accordingly, the model - in the late phase - is suitable to investigate alloantigen-independent factors of CAN and lacks markers of alloantigen-dependent processes.


Cross-reactions between Legionella pneumophila (serogroup 1) and twenty-eight other bacterial species, including other members of the family Legionellaceae.
Cross-reactions between Legionella pneumophila serogroup 1 and 28 other bacterial species were studied by various quantitative immunoelectrophoretic techniques. A sonicated L. pneumophila antigen and purified homologous rabbit antibody were used as a reference system. Few antigens (0 to 6) cross-reacted with non-Legionellaceae, but two were found in nearly all gram-negative bacteria tested (antigens no. 1 and 66). Antigen no. 66 of the L. pneumophila reference system was shown to be antigenically similar to the "common antigen" of Pseudomonas aeruginosa reported in many gram-negative bacteria. Greater than 85% of the antigens from L. pneumophila serogroup 1 cross-reacted with the other six serogroups of L. pneumophila. By contrast, Fluoribacter (Legionella) bozemanae, F. (L.) dumoffii, F. (L.) gormanii, and Tatlockia (Legionella) micdadei cross-reacted with only 45, 53, 39, and 43% of the reference system antigens, respectively. The antigenic relatedness of members of the Legionellaceae, expressed as a matching coefficient, is discussed in terms of its taxonomic significance. Serogroup-, genus-, and family-specific antigens are identified in the L. pneumophila reference system.


Implementation and characterization of flow injection in dissolution dynamic nuclear polarization NMR spectroscopy.
The use of dissolution dynamic nuclear polarization (D-DNP) offers substantially increased signals in liquid-state NMR spectroscopy. A challenge in realizing this potential lies in the transfer of the hyperpolarized sample to the NMR detector without loss of hyperpolarization. Here, the use of a flow injection method using high-pressure liquid leads to improved performance compared to the more common gas-driven injection, by suppressing residual fluid motions during the NMR experiment while still achieving a short injection time. Apparent diffusion coefficients are determined from pulsed field gradient echo measurements, and are shown to fall below 1.5 times the value of a static sample within 0.8 s. Due to the single-scan nature of D-DNP, pulsed field gradients are often the only choice for coherence selection or encoding, but their application requires stationary fluid. Sample delivery driven by a high-pressure liquid will improve the applicability of these types of D-DNP advanced experiments.


Amino acids protect epithelial cells from local toxicity by absorption enhancer, sodium laurate.
To develop the safe absorption-enhancing formulation attenuating the local toxicity caused by an absorption enhancer, sodium laurate (C12), the effects of amino acids on the local toxicity by C12 were examined in rats. The absorption of phenol red, an unabsorbable marker drug, was significantly enhanced by 10 mM C12 in an in situ colon loop study and the addition of L-glutamine (L-Gln), L-arginine, or L-methionine at 10 mM did not change the promoting effect of C12. However, C12 significantly increased the elution of phospholipids, total protein, and lactate dehydrogenase, which are markers for local toxicity, from colon, but these amino acids attenuated the local toxicity caused by C12 significantly. Transport study using an Ussing-type chamber showed that the permeability of colonic membrane to phenol red was significantly enhanced by C12 and that L-Gln did not decrease the permeability enhanced by C12. Transmucosal electrical resistance was extensively decreased by C12, indicating that C12 could enhance the drug absorption at least partly by expanding the paracellular route. L-Gln significantly, but not completely, recovered resistance lowered by C12. Electrical potential difference was markedly reduced by C12, suggesting that C12 lowered the viability of mucosal cells, but 10 mM L-Gln significantly recovered potential difference almost to the control level. These results suggested the possibility that absorption-enhancing formulation with low local toxicity, which is low enough to be used practically, could be developed by using an amino acid like L-Gln as an ingredient attenuating the local toxicity caused by C12.


In-situ ultra-sensitive infrared absorption spectroscopy of biomolecule interactions in real time with plasmonic nanoantennas.
Infrared absorption spectroscopy is a powerful biochemical analysis tool as it extracts detailed molecular structural information in a label-free fashion. Its molecular specificity renders the technique sensitive to the subtle conformational changes exhibited by proteins in response to a variety of stimuli. Yet, sensitivity limitations and the extremely strong absorption bands of liquid water severely limit infrared spectroscopy in performing kinetic measurements in biomolecules' native, aqueous environments. Here we demonstrate a plasmonic chip-based technology that overcomes these challenges, enabling the in-situ monitoring of protein and nanoparticle interactions at high sensitivity in real time, even allowing the observation of minute volumes of water displacement during binding events. Our approach leverages the plasmonic enhancement of absorption bands in conjunction with a non-classical form of internal reflection. These features not only expand the reach of infrared spectroscopy to a new class of biological interactions but also additionally enable a unique chip-based technology.


Pazopanib for treatment of advanced extraskeletal myxoid chondrosarcoma: a multicentre, single-arm, phase 2 trial.
Extraskeletal myxoid chondrosarcoma is a rare sarcoma with low sensitivity to cytotoxic chemotherapy. Retrospective evidence suggests that antiangiogenic drugs could be a treatment option. We aimed to investigate the activity of pazopanib, an antiangiogenic drug, in patients with advanced extraskeletal myxoid chondrosarcoma. In this single-arm, open-label phase 2 trial, three parallel independent cohorts of different histotypes of advanced sarcomas were recruited (extraskeletal myxoid chondrosarcoma, typical solitary fibrous tumour, and malignant-dedifferentiated solitary fibrous tumour). In each cohort, patients received pazopanib. In this Article, we report the results of the cohort of patients with advanced extraskeletal myxoid chondrosarcoma. Separate reporting of the three cohorts was prespecified in the study protocol. In this cohort, adult patients (aged ≥18 years) with a diagnosis of NR4A3-translocated, metastatic, or unresectable extraskeletal myxoid chondrosarcoma, who had Response Evaluation Criteria in Solid Tumors (RECIST) progression in the previous 6 months, and had an Eastern Cooperative Oncology Group performance status of 0-2, were enrolled at 11 study sites of the Spanish, Italian, and French sarcoma groups. Patients received oral pazopanib (800 mg/day) continuously, until disease progression, unacceptable toxicity, death, non-compliance, patient refusal, or investigator's decision. The primary endpoint was the proportion of patients achieving an objective response according to RECIST 1·1 in the modified intention-to-treat population (patients who provided consent and had a central molecularly confirmed diagnosis of extraskeletal myxoid chondrosarcoma). The safety analysis included all patients who received at least one dose of pazopanib. This study is registered with ClinicalTrials.gov, number NCT02066285. Between June 24, 2014, and Jan 17, 2017, 26 patients entered the study and started pazopanib. Of these, 23 met the eligibility criteria for the modified intention-to-treat analysis. Median follow-up was 27 months (IQR 18-30). 22 patients (one patient died before the primary analysis) were evaluable for the primary endpoint: four (18% [95% CI 1-36]) had a RECIST objective response. No deaths or grade 4 adverse events occurred. The most frequent grade 3 adverse events were hypertension (nine [35%] of 26 patients), increased concentration of alanine aminotransferase (six [23%]), and increased aspartate aminotransferase (five [19%]). Pazopanib had clinically meaningful antitumour activity in patients with progressive and advanced extraskeletal myxoid chondrosarcoma, and could be considered a suitable option after failure to respond to first-line anthracycline-based chemotherapy in these patients. Spanish Group for Research on Sarcomas, Italian Sarcoma Group, French Sarcoma Group, GlaxoSmithKline, and Novartis.


Biological dosimetry by the triage dicentric chromosome assay: potential implications for treatment of acute radiation syndrome in radiological mass casualties.
Biological dosimetry is an essential tool for estimating radiation dose. The dicentric chromosome assay (DCA) is currently the tool of choice. Because the assay is labor-intensive and time-consuming, strategies are needed to increase throughput for use in radiation mass casualty incidents. One such strategy is to truncate metaphase spread analysis for triage dose estimates by scoring 50 or fewer metaphases, compared to a routine analysis of 500 to 1000 metaphases, and to increase throughput using a large group of scorers in a biodosimetry network. Previously, the National Institutes for Allergies and Infectious Diseases (NIAID) and the Armed Forces Radiobiology Research Institute (AFRRI) sponsored a double-blinded interlaboratory comparison among five established international cytogenetic biodosimetry laboratories to determine the variability in calibration curves and in dose measurements in unknown, irradiated samples. In the present study, we further analyzed the published data from this previous study to investigate how the number of metaphase spreads influences dose prediction accuracy and how this information could be of value in the triage and management of people at risk for the acute radiation syndrome (ARS). Although, as expected, accuracy decreased with lower numbers of metaphase spreads analyzed, predicted doses by the laboratories were in good agreement and were judged to be adequate to guide diagnosis and treatment of ARS. These results demonstrate that for rapid triage, a network of cytogenetic biodosimetry laboratories can accurately assess doses even with a lower number of scored metaphases.


Efficacy of Fungicides and Rotational Programs for Management of Powdery Mildew on Cantaloupe.
Powdery mildew of cucurbits, caused by Podosphaera xanthii (syn. Sphaerotheca fuliginia auct. p.p. (Schltdl.) Pollacci), is a common and often severe disease in most areas of the world. Field trials were conducted with cantaloupe to compare disease management success provided by conventional fungicides and biofungicides having different inherent efficacies and modes of action, when applied alone throughout the treatment period or as components of fungicide application programs. Additionally, the portion of total disease control provided by each component fungicide within selected rotational programs was determined. When applied alone throughout the treatment period, disease severity in 2008 and 2009 compared with nontreated plants was reduced by values of 100, 99.3, and 98.1% by wettable sulfur (Microthiol Disperss), triflumizole (Procure), and quinoxyfen (Quintec), respectively; 83.9, 76.4, and 57.4% by trifloxystrobin (Flint), pyraclostrobin (Cabrio), or azoxystrobin (Quadris), respectively; and 39.8, 31.1, 30.0, and 28.6% by thiophanate-methyl (Topsin M), potassium bicarbonate (Kaligreen), kresoxim-methyl (Sovran), and Bacillus subtilis (Serenade), respectively. Rotational application programs composed of Microthiol Disperss, Procure, and Quintec reduced powdery mildew severity on cantaloupe by 97.5 to 100% in both trials. In comparison, disease reduction of 86.0 to 100% was achieved when the first and third fungicide applications were Quintec or Procure and the second and fourth applications were Cabrio, Flint, Kaligreen, Quadris, Serenade, Sovran, or Topsin M. In field trials designed to elucidate the portion of total disease control provided by each component fungicide within a rotational program, application sequences of Procure, Streptomyces lydicus (Actinovate), Procure, and Actinovate or Procure, Kaligreen, Procure, and Kaligreen resulted in reductions in powdery mildew severity of 69.1 and 78.7%, respectively. In comparison, inclusion of only the two Procure applications brought about a mean disease reduction of 85%, whereas inclusion of only the two Actinovate or Kaligreen applications reduced the level of powdery mildew control to 17.6 and 12.9%, respectively. The usefulness of fungicides with low inherent efficacy as resistance management partners when applied with highly efficacious at-risk fungicides may be questioned; however, the importance of any fungicide as a resistance management partner should be measured by its ability to dilute the selection pressure of the at-risk fungicide and to inhibit the growth of any resistant biotypes that may arise, factors which may not be reflected by its inherent efficacy in controlling disease. Because less efficacious fungicides did not increase the overall level of disease control, encouraging growers and pest control advisors to incorporate them into powdery mildew treatment programs may be a challenge, because resistance management is not their top priority. An ongoing educational effort emphasizing the benefits of resistance management programs with respect to prolonging the effectiveness of single-site mode of action fungicides is essential.


Synthesis and evaluation of the cytotoxicities of tetraindoles: observation that the 5-hydroxy tetraindole (SK228) induces G₂ arrest and apoptosis in human breast cancer cells.
Current chemical and biological interest in indole-3-carbinol (I3C) and its metabolites has resulted in the discovery of new biologically active indoles. As part of a program aimed at the development of indole analogues, tetraindoles 1-15 were prepared and their antiproliferative effects on human breast cancer cells were evaluated. The results show that the 5-hydroxy-tetraindole 8 (SK228) has optimum antiproliferative activity against breast adenocarcinoma (MCF 7 and MDA-MB-231) cells and that this activity involves G(2)-phase arrest of the cell cycle with a distinctive increase in the expression of cyclin B1 and phospho-cdc2. Further observations suggest that 5-hydroxy-tetraindole 8 induces apoptosis through externalization of membrane phosphatidylserine, DNA fragmentation, and activation of caspase-3. Given the fact that I3C and its metabolites have been shown to improve therapeutic efficacy and to have a broad range of antitumor activities in human cancer cells, the current findings have important pharmacological relevance as they open a promising route to the development of a potential chemotherapeutic application of tetraindoles as agents for the treatment of breast cancer.


Long-term effects of laparoscopic sleeve gastrectomy, gastric bypass, and adjustable gastric banding on type 2 diabetes.
This study aimed to compare the efficacy of laparoscopic sleeve gastrectomy (SG) with that of laparoscopic gastric bypass (GBP) and laparoscopic adjustable gastric banding (AGB) for glucose homeostasis in morbidly obese subjects with type 2 diabetes mellitus (T2DM) at a 3-year follow-up assessment and to elucidate the role of weight loss in the T2DM resolution after SG. For this study, 60 morbidly obese T2DM patients (44 females and 16 males) who underwent AGB (24 patients), GBP (16 patients), or SG (20 patients) between 1996 and 2008 were retrospectively analyzed. Age, sex, body mass index (BMI), estimated weight loss (EWL), fasting glycemia, HbA1c, euglycemic hyperinsulinemic clamp, discontinuation of diabetes treatment, and time until interruption of therapy were evaluated. In the study, 54 patients received oral hypoglycemic agents for at least 12 months before surgery, and 6 patients received insulin. The mean follow-up period was 36 months. The resolution rate was 60.8% for the AGB patients, 81.2% for the GBP patients, and 80.9% for the SG patients. The postoperative time until interruption of therapy was 12.6 months for the AGB patients, 3.2 months for the GBP patients, and 3.3 months for the SG patients. The hyperinsulinemic euglycemic clamp test was performed 12 months after surgery for the cured patients. Insulin resistance was restored to normal values in all the patients. The greatest improvement from preoperative values occurred in the SG group. For the not-cured GBP and SG patients, an improvement of 120 mg/dl in fasting plasma glucose was observed 3 months after the surgery, suggesting an enhancement in insulin sensitivity, which determines better medical control. The resolution rate remained constant at the 36-month follow-up evaluation in both the GBP and SG groups. All three bariatric procedures are effective in treating diabetes, with a 3-year follow-up evaluation showing an effect that lasts. The AGB procedure was the least effective. The antidiabetic effect was similarly precocious after GBP and SG compared with AGB. This difference may indicate that a hormonal mechanism may be involved, independent of weight loss.


Amino Acid Permeases and Virulence in Cryptococcus neoformans.
Fungal opportunistic pathogens colonize various environments, from plants and wood to human and animal tissue. Regarding human pathogens, one great challenge during contrasting niche occupation is the adaptation to different conditions, such as temperature, osmolarity, salinity, pressure, oxidative stress and nutritional availability, which may constitute sources of stress that need to be tolerated and overcome. As an opportunistic pathogen, C. neoformans faces exactly these situations during the transition from the environment to the human host, encountering nutritional constraints. Our previous and current research on amino acid biosynthetic pathways indicates that amino acid permeases are regulated by the presence of the amino acids, nitrogen and temperature. Saccharomyces cerevisiae and Candida albicans have twenty-four and twenty-seven genes encoding amino acid permeases, respectively; conversely, they are scarce in number in Basidiomycetes (C. neoformans, Coprinopsis cinerea and Ustilago maydis), where nine to ten permease genes can be found depending on the species. In this study, we have demonstrated that two amino acid permeases are essential for virulence in C. neoformans. Our data showed that C. neoformans uses two global and redundant amino acid permeases, Aap4 and Aap5 to respond correctly to thermal and oxidative stress. Double deletion of these permeases causes growth arrest in C. neoformans at 37°C and in the presence of hydrogen peroxide. The inability to uptake amino acid at a higher temperature and under oxidative stress also led to virulence attenuation in vivo. Our data showed that thermosensitivity caused by the lack of permeases Aap4 and Aap5 can be remedied by alkaline conditions (higher pH) and salinity. Permeases Aap4 and Aap5 are also required during fluconazole stress and they are the target of the plant secondary metabolite eugenol, a potent antifungal inhibitor that targets amino acid permeases. In summary, our work unravels (i) interesting physiological property of C. neoformans regarding its amino acid uptake system; (ii) an important aspect of virulence, which is the need for amino acid permeases during thermal and oxidative stress resistance and, hence, host invasion and colonization; and (iii) provides a convenient prototype for antifungal development, which are the amino acid permeases Aap4/Aap5 and their inhibitor.


In situ three-dimensional characterization of membrane fouling by protein suspensions using multiphoton microscopy.
Fouling of microfiltration membranes leads to severe flux declines and the need to clean or replace the membrane. In situ 3D characterization of protein fouling both on the surface and within the pores of the membrane was achieved using multiphoton microscopy. Time-lapse images of the fouled membrane were obtained for single suspensions and mixtures of fluorescently labeled bovine serum albumin and ovalbumin. Deposited protein aggregates were visible on the membrane and evidently play an important role in fouling. A combination of 3D images and resistance versus time data was used to identify the dominant fouling mechanism. Fouling is initially internally dominated, but after 1 and 15 min for ovalbumin and bovine serum albumin, respectively, the fouling becomes externally dominated. This is in good agreement with two-stage protein fouling models.


p32/gC1qR is indispensable for fetal development and mitochondrial translation: importance of its RNA-binding ability.
p32 is an evolutionarily conserved and ubiquitously expressed multifunctional protein. Although p32 exists at diverse intra and extracellular sites, it is predominantly localized to the mitochondrial matrix near the nucleoid associated with mitochondrial transcription factor A. Nonetheless, its function in the matrix is poorly understood. Here, we determined p32 function via generation of p32-knockout mice. p32-deficient mice exhibited mid-gestation lethality associated with a severe developmental defect of the embryo. Primary embryonic fibroblasts isolated from p32-knockout embryos showed severe dysfunction of the mitochondrial respiratory chain, because of severely impaired mitochondrial protein synthesis. Recombinant p32 binds RNA, not DNA, and endogenous p32 interacts with all mitochondrial messenger RNA species in vivo. The RNA-binding ability of p32 is well correlated with the mitochondrial translation. Co-immunoprecipitation revealed the close association of p32 with the mitoribosome. We propose that p32 is required for functional mitoribosome formation to synthesize proteins within mitochondria.


Neural resources for processing language and environmental sounds: evidence from aphasia.
Although aphasia is often characterized as a selective impairment in language function, left hemisphere lesions may cause impairments in semantic processing of auditory information, not only in verbal but also in nonverbal domains. We assessed the 'online' relationship between verbal and nonverbal auditory processing by examining the ability of 30 left hemisphere-damaged aphasic patients to match environmental sounds and linguistic phrases to corresponding pictures. The verbal and nonverbal task components were matched carefully through a norming study; 21 age-matched controls and five right hemisphere-damaged patients were also tested to provide further reference points. We found that, while the aphasic groups were impaired relative to normal controls, they were impaired to the same extent in both domains, with accuracy and reaction time for verbal and nonverbal trials revealing unusually high correlations (r = 0.74 for accuracy, r = 0.95 for reaction time). Severely aphasic patients tended to perform worse in both domains, but lesion size did not correlate with performance. Lesion overlay analysis indicated that damage to posterior regions in the left middle and superior temporal gyri and to the inferior parietal lobe was a predictor of deficits in processing for both speech and environmental sounds. The lesion mapping and further statistical assessments reliably revealed a posterior superior temporal region (Wernicke's area, traditionally considered a language-specific region) as being differentially more important for processing nonverbal sounds compared with verbal sounds. These results suggest that, in most cases, processing of meaningful verbal and nonverbal auditory information break down together in stroke and that subsequent recovery of function applies to both domains. This suggests that language shares neural resources with those used for processing information in other domains.


Award for Distinguished Contributions to Research in Public Policy.
The Award for Distinguished Contributions to Research in Public Policy is given to a psychologist who has made a distinguished empirical and/or theoretical contribution to research in public policy, either through a single extraordinary achievement or a lifetime of work. This contribution may consist of such factors as research leading others to view specific national policies differently; research demonstrating the importance of the application of psychological methods and theory to public policy; or research clarifying the ways in which scientific knowledge of human behavior informs public policy. The 2020 recipients of the APA Awards for Distinguished Contributions to Research in Public Policy were selected by the 2019 Committee on Psychology in the Public Interest Awards. (PsycInfo Database Record (c) 2020 APA, all rights reserved).


Properties and crystal structure of a beta-barrel folding mutant.
A mutant of a beta-barrel protein, rat intestinal fatty acid binding protein, was predicted to be more stable than the wild-type protein due to a novel hydrogen bond. Equilibrium denaturation studies indicated the opposite: the V60N mutant protein was less stable. The folding transitions followed by CD and fluorescence were reversible and two-state for both mutant and wild-type protein. However, the rates of denaturation and renaturation of V60N were faster. During unfolding, the initial rate was associated with 75-80% of the fluorescence and all of the CD amplitude change. A subsequent rate accounted for the remaining fluorescence change for both proteins; thus the intermediate state lacked secondary structure. During folding, one rate was detected by both fluorescence and CD after an initial burst phase for both wild-type and mutant. An additional slower folding rate was detected by fluorescence for the mutant protein. The structure of the V60N mutant has been obtained and is nearly identical to prior crystal structures of IFABP. Analysis of mean differences in hydrogen bond and van der Waals interactions did not readily account for the stability loss due to the mutation. However, significant average differences of the solvent accessible surface and crystallographic displacement factors suggest entropic destabilization.


CD27 expression discriminates between regulatory and non-regulatory cells after expansion of human peripheral blood CD4+ CD25+ cells.
It is clear that regulatory T cells (Treg) have an important role in preventing autoimmunity and modulating responses to pathogens. Full characterization of Treg cell function in human patients would be greatly facilitated by practical methods for expanding Treg in vitro. Methods for expansion have been reported but whether expression of surface and intracellular markers associated with freshly isolated Treg following expansion correlates with the maintenance of function is unclear. Our aim was to investigate the various methods of expansion and to correlate regulatory activity with expression of these markers. We show that, of the markers associated with freshly isolated Treg, only CD27 expression correlated with regulatory activity and could be used to isolate cells with regulatory activity from lines expanded from CD4+ CD25+ cells. Also, cells expressing high levels of the transcription factor forkhead box P3 (Foxp3) were confined to the CD27+ population within these lines. Expression of CD27 by cells in lines expanded from CD4+ CD25- cells varied depending on the stimulus used for expansion, but these lines did not have significant regulatory activity even when the CD27+ cells were tested. Analysis of synovial CD4+ CD25+ cells from reactive arthritis patients revealed that they were predominantly CD27 positive. This also applied to CD25(high) and CD25(intermediate) CD4+ cells, despite their reported different abilities to regulate. We conclude that, whilst CD27 is useful for identifying Treg in the cell lines obtained after expansion of CD4+ CD25+ cells, its expression may not reliably identify the Treg cell population in other T-cell populations such as those found in joints.


Modeling capping protein FRAP and CALI experiments reveals in vivo regulation of actin dynamics.
To gain insights on cellular mechanisms regulating actin polymerization, we used the Virtual Cell to model fluorescence recovery after photobleaching (FRAP) and chromophore-assisted laser inactivation (CALI) experiments on EGFP-capping protein (EGFP-CP). Modeling the FRAP kinetics demonstrated that the in vivo rate for the dissociation of CP from actin filaments is much faster (approximately 0.1 s(-1)) than that measured in vitro (0.01-0.0004 s(-1)). The CALI simulation revealed that in order to induce sustainable changes in cell morphology after CP inactivation, the cells should exhibit anticapping ability. We included the VASP protein as the anticapping agent in the modeling scheme. The model predicts that VASP affinity for barbed ends has a cooperative dependence on the concentration of VASP-barbed end complexes. This dependence produces a positive feedback that stabilizes the complexes and allows sustained growth at clustered filament tips. We analyzed the range of laser intensities that are sufficient to induce changes in cell morphology. This analysis demonstrates that FRAP experiments with EGFP-CP can be performed safely without changes in cell morphology, because, the intensity of the photobleaching beam is not high enough to produce the critical concentration of free barbed ends that will induce filament growth before diffusional replacement of EGFP-CP occurs.


Nonoptimal Codon Usage Is Critical for Protein Structure and Function of the Master General Amino Acid Control Regulator CPC-1.
Under amino acid starvation conditions, eukaryotic organisms activate a general amino acid control response. In Neurospora crassa, Cross Pathway Control Protein 1 (CPC-1), the ortholog of the Saccharomyces cerevisiae bZIP transcription factor GCN4, functions as the master regulator of the general amino acid control response. Codon usage biases are a universal feature of eukaryotic genomes and are critical for regulation of gene expression. Although codon usage has also been implicated in the regulation of protein structure and function, genetic evidence supporting this conclusion is very limited. Here, we show that Neurospora cpc-1 has a nonoptimal NNU-rich codon usage profile that contrasts with the strong NNC codon preference in the genome. Although substitution of the cpc-1 NNU codons with synonymous NNC codons elevated CPC-1 expression in Neurospora, it altered the CPC-1 degradation rate and abolished its amino acid starvation-induced protein stabilization. The codon-manipulated CPC-1 protein also exhibited different sensitivity to limited protease digestion. Furthermore, CPC-1 functions in rescuing the cell growth of the cpc-1 deletion mutant and activation of the expression of its target genes were impaired by the synonymous codon changes. Together, these results reveal the critical role of codon usage in regulation of CPC-1 expression and function and establish a genetic example of the importance of codon usage in protein folding.IMPORTANCE The general amino acid control response is critical for adaptation of organisms to amino acid starvation conditions. The preference to use certain synonymous codons is a universal feature of all genomes. Synonymous codon changes were previously thought to be silent mutations. In this study, we showed that the Neurospora cpc-1 gene has an unusual codon usage profile compared to other genes in the genome. We found that codon optimization of the cpc-1 gene without changing its amino acid sequence resulted in elevated CPC-1 expression, an altered protein degradation rate, and impaired protein functions due to changes in protein structure. Together, these results reveal the critical role of synonymous codon usage in regulation of CPC-1 expression and function and establish a genetic example of the importance of codon usage in protein structure.


Using cpsA-cpsB sequence polymorphisms and serotype-/group-specific PCR to predict 51 Streptococcus pneumoniae capsular serotypes.
Streptococcus pneumoniae polysaccharide and protein-conjugate vaccines are available against the most commonly isolated pneumococcal serotypes. Ongoing surveillance of invasive pneumococcal disease is needed in order to monitor changes in distribution of serotypes. Based on previously published sequences of capsular polysaccharide synthesis (cps) gene clusters of 16 pneumococcal serotypes, a molecular capsular typing (MCT) system has been developed, based on a combination of partial cpsA-cpsB sequencing and serotype- or serogroup-specific PCR, targeting the genes wzy and wzx (except for serotype 3). In this study, 151 S. pneumoniae isolates of known serotype (representing 51 serotypes) and 276 recent clinical isolates were used to develop MCT and compare it with conventional serotyping (CS) (total 427 isolates). On the basis of 376 heterogeneity sites in the cpsA-cpsB region, 89 sequence types (ST) were identified, of which 76 corresponded to a single serotype and 11 contained two serotypes. The correct serotypes in two of the latter (10A-23F-g and 23F-23A) were identified using serotype 23F-specific PCR. Limited CS was required for 92 (22 %) isolates to distinguish between the two serotypes in the nine other mixed ST (6A-6B-g, 6A-6B-q, 15B-22F, 33F-33A, 17F-35B, 18B-18C, 13-20, 25F-38, 31-42). MCT is a specific, objective and practical method that can predict the serotype of most S. pneumoniae isolates; it will facilitate epidemiological studies. Further study of the relationship between MCT and CS is needed in order to improve our understanding of serotype differentiation and to improve MCT methods further.


The anti-apoptotic protein lifeguard is expressed in breast cancer cells and tissues.
Lifeguard (LFG) is an anti-apoptotic protein that inhibits Fas-mediated death in tumour cells. However, the molecular function of human LFG in the carcinogenesis of human breast cells is uncertain. We studied the expression and function of endogenous LFG in four breast cancer cell lines (MCF-7, MDA-MB-231, T-47D and HS 578T), a human breast epithelial cell line (HS 578Bst), and in healthy and cancerous breast tissues. Molecular (Western blot and RT-PCR) and immunohistochemical techniques were used to investigate the LFG expression. To investigate the breast cancer cell proliferation in the presence of Fas, we performed fluorescent cell viability assays. The possible association of Fas with LFG was analyzed by immunofluorescence microscopy. In this paper, we provide convincing evidence that LFG is overexpressed in several human breast cancer cell lines. More importantly, we found that the LFG expression correlates with high tumour grades in primary breast tumours. Finally, we demonstrated that Fas sensitivity is reduced in breast cancer cell lines expressing LFG. Our results indicated that LFG is strongly expressed in breast cancer epithelial cells. Moreover, the overexpression of LFG correlated with tumour grade and reduced Fas sensitivity. Our findings support the idea that LFG may have a role in the downregulation of apoptosis in breast cancer cells.


Discovery of Novel BRD4 Ligand Scaffolds by Automated Navigation of the Fragment Chemical Space.
Fragment-based drug discovery (FBDD) is a very effective hit identification method. However, the evolution of fragment hits into suitable leads remains challenging and largely artisanal. Fragment evolution is often scaffold-centric, meaning that its outcome depends crucially on the chemical structure of the starting fragment. Considering that fragment screening libraries cover only a small proportion of the corresponding chemical space, hits should be seen as probes highlighting privileged areas of the chemical space rather than actual starting points. We have developed an automated computational pipeline to mine the chemical space around any specific fragment hit, rapidly finding analogues that share a common interaction motif but are structurally novel and diverse. On a prospective application on the bromodomain-containing protein 4 (BRD4), starting from a known fragment, the platform yields active molecules with nonobvious scaffold changes. The procedure is fast and inexpensive and has the potential to uncover many hidden opportunities in FBDD.


Characterisation of chemosensory trigeminal receptors in the rainbow trout, Oncorhynchus mykiss: responses to chemical irritants and carbon dioxide.
Trigeminally innervated, mechanically sensitive chemoreceptors (M) were previously identified in rainbow trout, Oncorhynchus mykiss, but it is not known whether these receptors are responsive only to noxious, chemical irritants or have a general chemosensory function. This study aimed to characterise the stimulus-response properties of these receptors in comparison with polymodal nociceptors (P). Both P and M gave similar response profiles to acetic acid concentrations. The electrophysiological properties were similar between the two different afferent types. To determine whether the receptors have a nociceptive function, a range of chemical stimulants was applied to these receptors, including non-noxious stimuli such as ammonium chloride, bile, sodium bicarbonate and alarm pheromone, and potentially noxious chemical irritants such as acetic acid, carbon dioxide, low pH, citric acid, citric acid phosphate buffer and sodium chloride. Only irritant stimuli evoked a response, confirming their nociceptive function. All receptor afferents tested responded to carbon dioxide (CO(2)) in the form of mineral water or soda water. The majority responded to 1% acetic acid, 2% citric acid, citric acid phosphate buffer (pH 3) and 5.0 mol l(-1) NaCl. CO(2) receptors have been characterised in the orobranchial cavity and gill arches in fish; however, this is the first time that external CO(2) receptors have been identified on the head of a fish. Because the fish skin is in constant contact with the aqueous environment, contaminants with a low pH or hypercapnia may stimulate the nociceptive system in fish.


Multifrequency electron spin resonance study of the dynamics of spin labeled T4 lysozyme.
An extensive set of electron spin resonance spectra was obtained over a wide range of frequencies (9, 95, 170, and 240 GHz) and temperatures (2 to 32 degrees C) to explore the dynamic modes of nitroxide-labeled T4 lysozyme in solution. A commonly used nitroxide side chain (R1), or a methylated analogue with hindered internal motion (R2), was substituted for the native side chain at solvent-exposed helical sites, 72 or 131. The spectra at all four frequencies were simultaneously fit with the slowly relaxing local structure (SRLS) model. Good fits were achieved at all the temperatures. Two principle dynamic modes are included in the SRLS model, the global tumbling of the protein and the internal motion consisting of backbone fluctuations and side chain isomerizations. Three distinct spectral components were required for R1 and two for R2 to account for the spectra at all temperatures. One is a highly ordered and slow motional component, which is observed in the spectra of both R1 and R2; it may correspond to conformers stabilized by interaction with the protein surface. The fraction of this component decreases with increasing temperature and is more populated in the R2 spectra, possibly arising from stronger interaction of the nitroxide ring with the protein surface due to the additional methyl group. The other two components of R1 and the second component of R2 are characterized by fast anisotropic diffusion and relatively low ordering, most likely corresponding to conformers having little or no interactions with nearby residues. Ficoll of different concentrations was added to increase the solution viscosity, thereby slowing down the global tumbling of the protein. A significant effect of Ficoll on the internal motion of an immobilized component was apparent in R2 but not in R1. The ability of such multifrequency studies to separate the effects of faster internal modes of motion from slower overall motions is clearly demonstrated, and its utility in future studies is considered.


Differences in the Students' Perceptions on the Teaching of Neuroanatomy in a Medical Curriculum Organized by Disciplines and an Integrated Medical Curriculum.
On the subject of curriculum reform, most European medical schools are moving away from an educational approach consisting of discipline-based courses to an integrated curriculum. The aim of this study was to compare, in the Faculty of Medicine of the University of Porto, Portugal, the teaching of neuroanatomy in a medical curriculum organized by disciplines and in an integrated medical curriculum. Two hundred sixty one students who completed the Curricular Unit with a discipline-based approach (Neuroanatomy) and 202 students who completed it with an integrated approach (Morphophysiology of the Nervous System) were asked to complete a questionnaire on their perceptions about the Curricular Unit. Our study showed that students of the Curricular Unit with a discipline-based approach had higher grades and evaluated it higher than students who followed the integrated approach. However, it also showed that students' grades had a significant effect on the evaluation of the curricular unit, with students with higher grades evaluating higher than students with lower grades. Besides, the majority of the students of the Curricular Unit with an integrated approach appreciated this curriculum model and highlighted as a positive point the successful integration of contents covered in the three components of the curricular unit. The curriculum reform led to the integration of neuroanatomy with other disciplines and resulted in a reduction of the teaching hours, a redefinition of the syllabus contents and the students' learning objectives, the introduction of new educational methods and changes in the evaluation system. Our study could not prove conclusively the supremacy of one pedagogic approach to neuroanatomy over the other. Future initiatives to explore different pedagogical models in medical education are needed and should be of major concern to the medical faculty.


Decision-making under great uncertainty: environmental management in an era of global change.
Global change issues are complex and the consequences of decisions are often highly uncertain. The large spatial and temporal scales and stakes involved make it important to take account of present and potential consequences in decision-making. Standard approaches to decision-making under uncertainty require information about the likelihood of alternative states, how states and actions combine to form outcomes and the net benefits of different outcomes. For global change issues, however, the set of potential states is often unknown, much less the probabilities, effect of actions or their net benefits. Decision theory, thresholds, scenarios and resilience thinking can expand awareness of the potential states and outcomes, as well as of the probabilities and consequences of outcomes under alternative decisions.


Esophageal stenting for benign and malignant disease: European Society of Gastrointestinal Endoscopy (ESGE) Guideline - Update 2021.
1: ESGE recommends placement of partially or fully covered self-expandable metal stents (SEMSs) for palliation of malignant dysphagia over laser therapy, photodynamic therapy, and esophageal bypass.Strong recommendation, high quality evidence. 2 : ESGE recommends brachytherapy as a valid alternative, alone or in addition to stenting, in esophageal cancer patients with malignant dysphagia and expected longer life expectancy.Strong recommendation, high quality evidence. 3: ESGE recommends esophageal SEMS placement for sealing malignant tracheoesophageal or bronchoesophageal fistulas. Strong recommendation, low quality evidence. 4 : ESGE does not recommend SEMS placement as a bridge to surgery or before preoperative chemoradiotherapy because it is associated with a high incidence of adverse events. Other options such as feeding tube placement are preferable. Strong recommendation, low quality evidence. 5: ESGE recommends against the use of SEMSs as first-line therapy for the management of benign esophageal strictures because of the potential for adverse events, the availability of alternative therapies, and their cost. Strong recommendation, low quality evidence. 6: ESGE suggests consideration of temporary placement of self-expandable stents for refractory benign esophageal strictures. Weak recommendation, moderate quality evidence. 7: ESGE suggests that fully covered SEMSs be preferred over partially covered SEMSs for the treatment of refractory benign esophageal strictures because of their very low risk of embedment and ease of removability. Weak recommendation, low quality evidence. 8: ESGE recommends the stent-in-stent technique to remove partially covered SEMSs that are embedded in the esophageal wall. Strong recommendation, low quality evidence. 9: ESGE recommends that temporary stent placement can be considered for the treatment of leaks, fistulas, and perforations. No specific type of stent can be recommended, and the duration of stenting should be individualized. Strong recommendation, low quality of evidence. 10 : ESGE recommends considering placement of a fully covered large-diameter SEMS for the treatment of esophageal variceal bleeding refractory to medical, endoscopic, and/or radiological therapy, or as initial therapy for patients with massive bleeding. Strong recommendation, moderate quality evidence.


Proteomic adaptations to starvation prepare Escherichia coli for disinfection tolerance.
Despite the low nutrient level and constant presence of secondary disinfectants, bacterial re-growth still occurs in drinking water distribution systems. The molecular mechanisms that starved bacteria use to survive low-level chlorine-based disinfectants are not well understood. The objective of this study is to investigate these molecular mechanisms at the protein level that prepare starved cells for disinfection tolerance. Two commonly used secondary disinfectants chlorine and monochloramine, both at 1 mg/L, were used in this study. The proteomes of normal and starved Escherichia coli (K12 MG1655) cells were studied using quantitative proteomics. Over 60-min disinfection, starved cells showed significantly higher disinfection tolerance than normal cells based on the inactivation curves for both chlorine and monochloramine. Proteomic analyses suggest that starvation may prepare cells for the oxidative stress that chlorine-based disinfection will cause by affecting glutathione metabolism. In addition, proteins involved in stress regulation and stress responses were among the ones up-regulated under both starvation and chlorine/monochloramine disinfection. By comparing the fold changes under different conditions, it is suggested that starvation prepares E. coli for disinfection tolerance by increasing the expression of enzymes that can help cells survive chlorine/monochloramine disinfection. Protein co-expression analyses show that proteins in glycolysis and pentose phosphate pathway that were up-regulated under starvation are also involved in disinfection tolerance. Finally, the production and detoxification of methylglyoxal may be involved in the chlorine-based disinfection and cell defense mechanisms.


Gamma Oscillations Underlie the Maintenance of Feature-Specific Information and the Contents of Visual Working Memory.
Visual working memory (VWM) sustains information online as integrated object representations. Neuronal mechanisms supporting the maintenance of feature-specific information have remained unidentified. Synchronized oscillations in the gamma band (30-120 Hz) characterize VWM retention and predict task performance, but whether these oscillations are specific to memorized features and VWM contents or underlie general executive VWM functions is not known. In the present study, we investigated whether gamma oscillations reflect the maintenance of feature-specific information in VWM. Concurrent magneto- and electroencephalography was recorded while subjects memorized different object features or feature conjunctions in identical VWM experiments. Using a data-driven source analysis approach, we show that the strength, load-dependence, and source topographies of gamma oscillations in the visual cortex differentiate these memorized features. Load-dependence of gamma oscillations in feature-specific visual and prefrontal areas also predicts VWM accuracy. Furthermore, corroborating the hypothesis that gamma oscillations support the perceptual binding of feature-specific neuronal assemblies, we also show that VWM for color-location conjunctions is associated with stronger gamma oscillations than that for these features separately. Gamma oscillations hence support the maintenance of feature-specific information and reflect VWM contents. The results also suggest that gamma oscillations contribute to feature binding in the formation of memory representations.


Wannier90 as a community code: new features and applications.
Wannier90 is an open-source computer program for calculating maximally-localised Wannier functions (MLWFs) from a set of Bloch states. It is interfaced to many widely used electronic-structure codes thanks to its independence from the basis sets representing these Bloch states. In the past few years the development of Wannier90 has transitioned to a community-driven model; this has resulted in a number of new developments that have been recently released in Wannier90 v3.0. In this article we describe these new functionalities, that include the implementation of new features for wannierisation and disentanglement (symmetry-adapted Wannier functions, selectively-localised Wannier functions, selected columns of the density matrix) and the ability to calculate new properties (shift currents and Berry-curvature dipole, and a new interface to many-body perturbation theory); performance improvements, including parallelisation of the core code; enhancements in functionality (support for spinor-valued Wannier functions, more accurate methods to interpolate quantities in the Brillouin zone); improved usability (improved plotting routines, integration with high-throughput automation frameworks), as well as the implementation of modern software engineering practices (unit testing, continuous integration, and automatic source-code documentation). These new features, capabilities, and code development model aim to further sustain and expand the community uptake and range of applicability, that nowadays spans complex and accurate dielectric, electronic, magnetic, optical, topological and transport properties of materials.


Perioperative lidocaine infusion reduces the incidence of post-mastectomy chronic pain: a double-blind, placebo-controlled randomized trial.
Chronic post-surgical pain (CPSP) is a not uncommon complication after mastectomy, with a reported incidence between 20% and 68%. Careful dissection, the use of minimally invasive surgical techniques, and attempts to reduce the associated inflammatory and hyperalgesic responses are suggested methods to prevent CPSP. To determine if the use of perioperative lidocaine infusion is associated with decreased incidence of CPSP after mastectomy. Double-blind, placebo-controlled randomized trial. This is a secondary analysis of data from 61 out of 71 patients who underwent mastectomy for breast cancer. Patients were randomized to either placebo (Group P; n = 27) or intravenous lidocaine (Group L; n = 34, bolus 1.5 mg/kg at induction, then infusion at 2 mg/kg/hr, up to 2 hours after the end of surgery) in a prospective double-blind design. CPSP was assessed at 6 months after surgery. Stepwise logistic regression analysis was performed to assess the efficacy of lidocaine. Overall 12 (20%) patients developed CPSP, 8 (30%) in the placebo group and 4 (12%) in the lidocaine group. Predictive factors for CPSP that remained significant after multivariate analysis included lidocaine (associated with a 20-fold decrease in CPSP, P = 0.013), breast implant placement (associated with a 16-fold increase in CPSP, P = 0.034), and radiotherapy (associated with a 29-fold increase in CPSP, P = 0.008). Small sample size. Perioperative lidocaine administration was associated with a decreased incidence of CPSP, while breast implant placement and radiotherapy were associated with an increased incidence. These findings suggest a protective effect of lidocaine on CPSP development in mastectomy patients.


Apoptosis in inner ear sensory hair cells.
Apoptosis, or controlled cell death, is a normal part of cellular lifespan. Cell death of cochlear hair cells causes deafness; an apoptotic process that is not well understood. Worldwide, 1.3 billion humans suffer some form of hearing loss, while 360 million suffer debilitating hearing loss as a direct result of the absence of these cochlear hair cells (Worldwide Hearing, 2014). Much is known about apoptosis in other systems and in other cell types thanks to studies done since the mid-20th century. Here we review current literature on apoptosis in general, and causes of deafness and cochlear hair cells loss as a result of apoptosis. The family of B-cell lymphoma (Bcl) proteins are among the most studied and characterized. We will review current literature on the Bcl2 and Bcl6 protein interactions in relation to apoptosis and their possible roles in vulnerability and survival of cochlear hair cells.


Evaluation of Changes in Ghanaian Students' Attitudes Towards Science Following Neuroscience Outreach Activities: A Means to Identify Effective Ways to Inspire Interest in Science Careers.
The scientific capacity in many African countries is low. Ghana, for example, is estimated to have approximately twenty-three researchers per a million inhabitants. In order to improve interest in science among future professionals, appropriate techniques should be developed and employed to identify barriers and correlates of science education among pre-university students. Young students' attitudes towards science may affect their future career choices. However, these attitudes may change with new experiences. It is, therefore, important to evaluate potential changes in students' attitudes towards science after their exposure to experiences such as science outreach activities. Through this, more effective means of inspiring and mentoring young students to choose science subjects can be developed. This approach would be particularly beneficial in countries such as Ghana, where: (i) documented impacts of outreach activities are lacking; and (ii) effective means to develop scientist-school educational partnerships are needed. We have established an outreach scheme, aimed at helping to improve interaction between scientists and pre-university students (and their teachers). Outreach activities are designed and implemented by undergraduate students and graduate teaching assistants, with support from faculty members and technical staff. Through this, we aim to build a team of trainee scientists and graduates who will become ambassadors of science in their future professional endeavors. Here, we describe an approach for assessing changes in junior high school students' attitudes towards science following classroom neuroscience outreach activities. We show that while students tended to agree more with questions concerning their perceptions about science learning after the delivery of outreach activities, significant improvements were obtained for only two questions, namely "I enjoy science lessons" and "I want to be a scientist in the future." Furthermore, there was a generally strong trend towards a change in attitude for questions that sought information about students' perceptions about scientists (both positive and negative perceptions). In addition, outreach providers reported that their involvement in this public engagement scheme helped them acquire several transferable skills that will be beneficial in their studies and career development. These include vital skills in project and time management, teamwork and public speaking. Altogether, our findings provide novel indications that the development of scientist-school outreach partnerships in Ghana has valuable implications for science education and capacity development.


Insertion and removal of intrauterine devices.
The intrauterine device (IUD) is an effective contraceptive for many women. The copper-releasing IUD can be used for 10 years before replacement and is a good choice for women who cannot, or choose not to, use hormone-releasing contraceptives. However, some women experience an increase in menstrual blood loss and dysmenorrhea. The progestin-releasing IUD can be used for five years. It may reduce menorrhagia and dysmenorrhea, although some women have increased spotting and bleeding during the first months after insertion. The ideal candidates for IUD use are parous women in stable, monogamous relationships. Pregnancy, unexplained vaginal bleeding, and a lifestyle placing the woman at risk for sexually transmitted diseases are contraindications to IUD use. Insertion of the IUD can take place at any time during the menstrual cycle provided the woman is not pregnant. Before insertion, a bimanual examination and a sounding of the uterus are necessary to determine the uterus position and the depth of the uterine cavity. The IUD is inserted into the uterus according to individual protocols, with the threads cut at a length to allow the patient to check the device's position. Expulsion may occur with both types of IUDs.


Design of Gaussia luciferase-based bioluminescent stem-loop probe for sensitive detection of HIV-1 nucleic acids.
Here we describe the design of a bioluminescent stem-loop probe for the sensitive detection of HIV-1 spliced RNA. In this study, we employed Gaussia luciferase (GLuc), a bioluminescent protein that has several advantages over other bioluminescent proteins, including smaller size, higher bioluminescent intensity, and chemical and thermal stability. GLuc was chemically conjugated to the DABCYL-modified stem-loop probe (SLP) and was purified with a 2-step process to remove unconjugated GLuc and SLP. The binding of the target RNA to the loop region of the SLP results in the open conformation separating the stem part of SLP. GLuc conjugated to the stem acts as a reporter that produces light by a chemical reaction upon adding its substrate, coelenterazine in the presence of the target, while DABCYL serves as a quencher of bioluminescence in the closed conformation of SLP in the absence of the target. The optimized GLuc based-SLP assay resulted in a signal-to-background ratio of 47, which is the highest reported with bioluminescent SLPs and is significantly higher compared to traditional fluorescence-based SLPs that yield low signal to background ratio. Moreover, the assay showed an excellent selectivity against a single and double mismatched nucleic acid target, low detection limit, and ability to detect spiked HIV-1 RNA in human serum matrix.


Posttreatment motivation and alcohol treatment outcome 9 months later: findings from structural equation modeling.
To investigate the association between posttreatment motivation to change as measured by the Readiness to Change Questionnaire Treatment Version and drinking outcomes 9 months after the conclusion of treatment for alcohol problems. Data from 392 participants in the United Kingdom Alcohol Treatment Trial were used to fit structural equation models investigating relationships between motivation to change pre- and posttreatment and 5 outcomes 9 months later. The models included pathways through changes in drinking behavior during treatment and adjustment for sociodemographic information. Greater posttreatment motivation (being in action vs. preaction) was associated with 3 times higher odds of the most stringent definition of positive outcome (being abstinent or entirely a nonproblem drinker) 9 months later (odds ratio = 3.10, 95% confidence interval [1.83, 5.25]). A smaller indirect effect of pretreatment motivation on this outcome was seen from pathways through drinking behavior during treatment and posttreatment motivation (probit coefficient = 0.08, 95% confidence interval [0.03, 0.14]). A similar pattern of results was seen for other outcomes evaluated. Posttreatment motivation to change has hitherto been little studied and is identified here as a clearly important predictor of longer term treatment outcome.


Case Report: Co-existence of sarcoidosis and Takayasu arteritis.
Takayasu arteritis is a rare systemic large vessel vasculitis affecting the aorta and its branches. Sarcoidosis, too, is an inflammatory disease. Both entities are granulomatous conditions with a question of possible linkquestionable association in their etiopathogenesis. Only a few cases of their coexistence have been reported in the literature. To our knowledge, no such cases have been reported from Nepal. We report a Nepalese woman who presented with non-productive cough, progressive shortness of breath and chest tightness of 3 years duration. She had a history of recurrent bilateral granulomatous uveitis over the previous 3 years. Examination revealed clubbing of digits, absent pulses over the left radial, ulnar and brachial arteries, and a weak pulse over the right arm including the bilateral carotid arteries. Pulmonary function test showed restrictive pattern, a high-resolution computed tomography (HRCT) scan of the chest revealed findings suggestive of pulmonary sarcoidosis. A CT aortogram suggested large vessel vasculitis. Bronchoscopy with biopsy revealed granulomatous inflammation, negative for malignancy and tuberculosis. She was hence, diagnosed with co-existing Takayasu arteritis and sarcoidosis, and treated with Prednisolone 60 mg once daily with dramatic improvement over 4 days and was discharged stable on domiciliary oxygen. She is currently on azathioprine 50 mg, prednisolone 10 mg without the need for supplemental oxygen. This case report highlights the importance of a proper physical examination as a guide to the use of modern technology in making a correct diagnosis. Furthermore, in countries where tuberculosis is endemic, it should always come as the most important differential diagnosis of granulomatous inflammation.


Mini review on the structure and supramolecular assembly of VDAC.
The Voltage Dependent Anion Channel (VDAC) is the most abundant protein in the outer membrane of mitochondria. This strategic localization puts it at the heart of a great number of phenomena. Its recent implication in apoptosis is an example of the major importance of this protein and has created a surge of interest in VDAC. There is no atomic-resolution structure allowing a better understanding of the function of VDAC, so alternative techniques to X-ray diffraction have been used to study VDAC. Here we discuss structural models from folding predictions and review data acquired by Atomic Force Microscopy (AFM) imaging that allowed to observe VDAC's structure and supramolecular organization in the mitochondrial outer membrane.


Variation in response to artificial selection for light sensitivity in guppies (Poecilia reticulata).
We performed artificial selection on the visual system in guppies (Poecilia reticulata), using the optomotor reaction threshold as the selection criterion. Two lines were selected for increased sensitivity to blue light, two were selected for increased sensitivity to red light, and two were unselected controls. There was significant response to selection in all four selected lines and significant heritability for sensitivity. An examination of the spectral sensitivity function showed that the form of the response differed between the red and blue lines and among the red lines. Such divergence is likely because there are many different mechanisms allowing response to selection for spectral sensitivity. Diverse mechanisms allow a divergent response by different populations to the same selective pressures. Such a mechanism can promote diversity in vision and visual signals, and any multicomponent system where different components can respond to the same selective regime.


The population value of quality indicator reporting: a framework for prioritizing health care performance measures.
The Agency for Healthcare Research and Quality (AHRQ) National Healthcare Quality and Disparities Reports contain more than 250 quality indicators, such as whether a patient with a suspected heart attack received an aspirin. The Department of Health and Human Services National Quality Measures Clearinghouse identifies more than 2,100 such indicators. Because resources for making quality improvements are limited, there is a need to prioritize among these indicators. We propose an approach to assess how reporting specific quality indicators would change care to improve the length and quality of life of the US population. Using thirteen AHRQ quality indicators with readily available data on the benefits of indicator reporting, we found that seven of them account for 93 percent of total benefits, while the remaining six account for only 7 percent of total benefits. Use of a framework such as this could focus resources on indicators having the greatest expected impact on population health.


Descriptive epidemiology of osteoarthritis in British Columbia, Canada.
Osteoarthritis (OA) is a highly prevalent and often disabling disease. Data on the incidence of OA in the general population are limited. Our objectives were (1) to estimate OA prevalence and incidence rates by age and sex in a geographically defined population of 4 million people [British Columbia (BC), Canada] using an administrative database; and (2) to determine the effects of different administrative definitions of OA and observation (run-in) time on such estimates. We used data on all visits to health professionals and hospital admissions covered by the Medical Services Plan (MSP) of BC for the fiscal years 1991-92 through 2000-01. OA was defined based on International Classification of Diseases, 9th Revision, diagnostic codes required for administrative purposes. The overall prevalence of OA in 2001 was 10.8%: 8.9% in men and 12.6% in women. Prevalence was higher in women in all age groups. By age 70-74 years, about one-third of men and 40% of women had OA. Incidence rates in 2000-01 were 11.7 per 1000 person-years in the total population, 10.0 in men and 13.4 in women. Rates increased linearly with age between 50 and 80 years. Both prevalence and incidence depended strongly on the definition of OA and the run-in period. Prevalence of physician-diagnosed OA in BC was slightly lower than self-reported prevalence of arthritis in population surveys. Routinely collected administrative data could be a valuable source of information for OA surveillance, but more research is needed on the validity of OA diagnosis in administrative databases.