Pore-formation by adenylate cyclase toxoid activates dendritic cells to prime CD8+ and CD4+ T cells.
The adenylate cyclase toxin-hemolysin (CyaA) of Bordetella pertussis is a bi-functional leukotoxin. It penetrates myeloid phagocytes expressing the complement receptor 3 and delivers into their cytosol its N-terminal adenylate cyclase enzyme domain (~400 residues). In parallel, ~1300 residue-long RTX hemolysin moiety of CyaA forms cation-selective pores and permeabilizes target cell membrane for efflux of cytosolic potassium ions. The non-enzymatic CyaA-AC(-) toxoid, has repeatedly been successfully exploited as an antigen delivery tool for stimulation of adaptive T-cell immune responses. We show that the pore-forming activity confers on the CyaA-AC(-) toxoid a capacity to trigger Toll-like receptor and inflammasome signaling-independent maturation of CD11b-expressing dendritic cells (DC). The DC maturation-inducing potency of mutant toxoid variants in vitro reflected their specifically enhanced or reduced pore-forming activity and K(+) efflux. The toxoid-induced in vitro phenotypic maturation of DC involved the activity of mitogen activated protein kinases p38 and JNK and comprised increased expression of maturation markers, interleukin 6, chemokines KC and LIX and granulocyte-colony-stimulating factor secretion, prostaglandin E2 production and enhancement of chemotactic migration of DC. Moreover, i.v. injected toxoids induced maturation of splenic DC in function of their cell-permeabilizing capacity. Similarly, the capacity of DC to stimulate CD8(+) and CD4(+) T-cell responses in vitro and in vivo was dependent on the pore-forming activity of CyaA-AC(-). This reveals a novel self-adjuvanting capacity of the CyaA-AC(-) toxoid that is currently under clinical evaluation as a tool for delivery of immunotherapeutic anti-cancer CD8(+) T-cell vaccines into DC.


An optical authentication system based on imaging of excitation-selected lanthanide luminescence.
Secure data encryption relies heavily on one-way functions, and copy protection relies on features that are difficult to reproduce. We present an optical authentication system based on lanthanide luminescence from physical one-way functions or physical unclonable functions (PUFs). They cannot be reproduced and thus enable unbreakable encryption. Further, PUFs will prevent counterfeiting if tags with unique PUFs are grafted onto products. We have developed an authentication system that comprises a hardware reader, image analysis, and authentication software and physical keys that we demonstrate as an anticounterfeiting system. The physical keys are PUFs made from random patterns of taggants in polymer films on glass that can be imaged following selected excitation of particular lanthanide(III) ions doped into the individual taggants. This form of excitation-selected imaging ensures that by using at least two lanthanide(III) ion dopants, the random patterns cannot be copied, because the excitation selection will fail when using any other emitter. With the developed reader and software, the random patterns are read and digitized, which allows a digital pattern to be stored. This digital pattern or digital key can be used to authenticate the physical key in anticounterfeiting or to encrypt any message. The PUF key was produced with a staggering nominal encoding capacity of 73600. Although the encoding capacity of the realized authentication system reduces to 6 × 10104, it is more than sufficient to completely preclude counterfeiting of products.


mTORC1 Signaling and Negative Lens-Induced Axial Elongation.
The mechanism underlying axial elongation during myopia progression remains unknown. Epidermal growth factor receptor (EGFR) signaling is associated with axial elongation. We explored whether mammalian target of rapamycin complex 1 (mTORC1) signaling acts as the downstream pathway of EGFR and participates in negative lens-induced axial elongation (NLIAE). Three-week-old male pigmented guinea pigs underwent binocular NLIAE. (1) To investigate whether EGFR is the upstream regulator of mTORC1, an EGFR inhibitor (20 µg erlotinib) was intravitreally injected once a week for three weeks. (2) To assess the effect of mTORC1 inhibition on NLIAE, an mTORC1 inhibitor (2 µg, 10 µg, and 20 µg everolimus) was intravitreally injected once a week for three weeks. (3) To explore the long-term effect of mTORC1 overactivation on axial elongation, an mTORC1 agonist (4 µg MHY1485) was intravitreally injected once a week for three months. Biometric measurements included axial length and choroidal thickness were performed. Compared with the guinea pigs without NLIAE, NLIAE was associated with activation of mTORC1 signaling, which was suppressed by intravitreal erlotinib injection. Intravitreally injected everolimus suppressed NLIAE-induced axial elongation, mTORC1 activation, choroidal thinning, and hypoxia-inducible factor-1α expression in the sclera. Immunofluorescence revealed that the retinal pigment epithelium was the primary location of mTORC1 activation during NLIAE. Combining NLIAE and MHY1485 intravitreal injections significantly promoted axial elongation, choroidal thinning, and peripapillary choroidal atrophy. The mTORC1 signaling is associated with increased axial elongation, as in NLIAE, raising the possibility of inhibiting mTORC1 as a novel treatment for slowing myopia progression.


Whose quality of life is it anyway? Discrepancies between youth and parent health-related quality of life ratings in type 1 and type 2 diabetes.
Health-related quality of life (HRQOL) is a critical diabetes outcome, yet differences between youth and parent-proxy ratings can make interpretation difficult. This study aims to explore potential differences between self- and parent-reports of Pediatric Quality of Life Inventory (PedsQL) scores from youth with type 1 (T1D) or type 2 diabetes (T2D) and to evaluate associations between discrepancies, PedsQL scores, and glycemic control (HbA1c). Youth and parents in the SEARCH for Diabetes in Youth Study (T1D: age 5-18, n = 3402; T2D: age 8-18, n = 353) completed the PedsQL Generic and Diabetes Modules, and youth provided a blood sample to assess HbA1c. Discrepancies (youth minus parent PedsQL ratings) were calculated and examined by age and diabetes type, and associations with youth PedsQL scores and HbA1c were evaluated. Discrepancies existed between youth and parent-proxy reports of generic and diabetes PedsQL scores in T1D and T2D (all p values < 0.01). Higher (more favorable) ratings were reported by youth except for those 5-7-years old, where parents' scores were higher. When parent-proxy scores were higher, discrepancies were largest when the child reported low PedsQL scores. Higher HbA1c was associated with larger discrepancies (youth scores higher) for adolescents with T1D. Discrepant PedsQL ratings suggest that parents may often underestimate youths' HRQOL except in the youngest children. Although examining both reports is optimal, the youth report should be prioritized, particularly for young children with T1D and for adolescents with either T1D or T2D.


Leucine Modulates Mitochondrial Biogenesis and SIRT1-AMPK Signaling in C2C12 Myotubes.
Previous studies from this laboratory demonstrate that dietary leucine protects against high fat diet-induced mitochondrial impairments and stimulates mitochondrial biogenesis and energy partitioning from adipocytes to muscle cells through SIRT1-mediated mechanisms. Moreover, β-hydroxy-β-methyl butyrate (HMB), a metabolite of leucine, has been reported to activate AMPK synergistically with resveratrol in C2C12 myotubes. Therefore, we hypothesize that leucine-induced activation of SIRT1 and AMPK is the central event that links the upregulated mitochondrial biogenesis and fatty acid oxidation in skeletal muscle. Thus, C2C12 myotubes were treated with leucine (0.5 mM), alanine (0.5 mM), valine (0.5 mM), EX527 (SIRT1 inhibitor, 25 μM), and Compound C (AMPK inhibitor, 25 μM) alone or in combination to determine the roles of AMPK and SIRT1 in leucine-modulation of energy metabolism. Leucine significantly increased mitochondrial content, mitochondrial biogenesis-related genes expression, fatty acid oxidation, SIRT1 activity and gene expression, and AMPK phosphorylation in C2C12 myotubes compared to the controls, while EX527 and Compound C markedly attenuated these effects. Furthermore, leucine treatment for 24 hours resulted in time-dependent increases in cellular NAD(+), SIRT1 activity, and p-AMPK level, with SIRT1 activation preceding that of AMPK, indicating that leucine activation of SIRT1, rather than AMPK, is the primary event.


Accelerating changes in ice mass within Greenland, and the ice sheet's sensitivity to atmospheric forcing.
From early 2003 to mid-2013, the total mass of ice in Greenland declined at a progressively increasing rate. In mid-2013, an abrupt reversal occurred, and very little net ice loss occurred in the next 12-18 months. Gravity Recovery and Climate Experiment (GRACE) and global positioning system (GPS) observations reveal that the spatial patterns of the sustained acceleration and the abrupt deceleration in mass loss are similar. The strongest accelerations tracked the phase of the North Atlantic Oscillation (NAO). The negative phase of the NAO enhances summertime warming and insolation while reducing snowfall, especially in west Greenland, driving surface mass balance (SMB) more negative, as illustrated using the regional climate model MAR. The spatial pattern of accelerating mass changes reflects the geography of NAO-driven shifts in atmospheric forcing and the ice sheet's sensitivity to that forcing. We infer that southwest Greenland will become a major future contributor to sea level rise.


Flexible Lithium-Ion Fiber Battery by the Regular Stacking of Two-Dimensional Titanium Oxide Nanosheets Hybridized with Reduced Graphene Oxide.
Increasing interest has recently been devoted to developing small, rapid, and portable electronic devices; thus, it is becoming critically important to provide matching light and flexible energy-storage systems to power them. To this end, compared with the inevitable drawbacks of being bulky, heavy, and rigid for traditional planar sandwiched structures, linear fiber-shaped lithium-ion batteries (LIB) have become increasingly important owing to their combined superiorities of miniaturization, adaptability, and weavability, the progress of which being heavily dependent on the development of new fiber-shaped electrodes. Here, we report a novel fiber battery electrode based on the most widely used LIB material, titanium oxide, which is processed into two-dimensional nanosheets and assembled into a macroscopic fiber by a scalable wet-spinning process. The titania sheets are regularly stacked and conformally hybridized in situ with reduced graphene oxide (rGO), thereby serving as efficient current collectors, which endows the novel fiber electrode with excellent integrated mechanical properties combined with superior battery performances in terms of linear densities, rate capabilities, and cyclic behaviors. The present study clearly demonstrates a new material-design paradigm toward novel fiber electrodes by assembling metal oxide nanosheets into an ordered macroscopic structure, which would represent the most-promising solution to advanced flexible energy-storage systems.


Characterization of a novel molluskan tyrosine kinase receptor that inhibits neurite regeneration.
Receptor tyrosine kinases play many important roles in neuronal signaling including regulating neurite outgrowth. We have identified a novel receptor tyrosine kinase, neurite outgrowth regulating kinase (nork) from Aplysia californica. A fragment of this kinase was also identified in another mollusk, Lymnaea. The kinase domain is equally homologous to the Ret (rearranged during transformation) and fibroblast growth factor receptor families, but the extracellular domain is entirely novel, suggesting that it binds a nonconserved ligand. Overexpression of neurite outgrowth regulating kinase, but not a kinase dead form, causes a reduction in neurite outgrowth of Aplysia sensory neurons. Thus, we have identified a novel receptor tyrosine kinase implicated in regulating neurite outgrowth.


Syntheses, Structures, and Comparisons of Heterometallic Uranyl Iodobenzoates with Monovalent Cations.
The syntheses and crystal structures of six new heterometallic compounds containing the UO22+ cation, o-, m-, and p-iodobenzoic acid ligands, and Tl+, Rb+, and Cs+ cations which adopt the role of both charge balancing cation and secondary metal center are described, as are the luminescent properties for Tl+ containing compounds 1, 4, and 6. The structures of compounds 1-3 are isomorphous and contain uranyl monomers bound by o-iodobenzoic acid ligands with Tl+, Rb+, and Cs+ cations acting as secondary metal centers. Compounds 4 and 5 are also isomorphous and feature m-iodobenzoic acid ligands bound to the uranyl cation along with Tl+ and Rb+ cations. Compound 6 is unique in this series as it is assembled from a dimeric uranyl unit and features p-iodobenzoic acid ligands and Tl+ cations which function as charge balancing secondary metal centers. Single crystal X-ray diffraction analysis of these materials suggests that the secondary metal cations are incorporated based on the size of their ionic radius (Tl+ < Rb+ < Cs+), which is directly related to the size of the "pocket" observed in 1-6. Further, Voronoi-Dirichlet tessellation and Hirshfeld surface analysis were used to probe the coordination environment of the secondary metal centers as part of ongoing efforts to develop metrics for determining the coordination number of secondary metal cations in similar systems.


Inheritance of mutations in the V2 receptor gene in thirteen families with nephrogenic diabetes insipidus.
Congenital nephrogenic diabetes insipidus (NDI) is an X-linked recessive disease characterized by insensitivity of the distal nephron to the antidiuretic effect of arginine vasopressin. The hypothesis that the defect underlying NDI might be a dysfunctional renal vasopressin V2 receptor has recently been proven by the identification of mutations in the V2 receptor gene in NDI patients. We examined thirteen unrelated Dutch NDI families and identified thirteen distinct and unique mutations. These included nine missense mutations, two nonsense mutations and two small deletions and were found in the extracellular domains II, III and IV, the intracellular domains II and IV and in the transmembrane loops I, II, IV and V of the vasopressin type 2 receptor. In the families with multiple NDI patients the mutated gene cosegregated with the disease. Our data suggest a higher mutation frequency in male than in female gametes. No discrepancies between carrier detection by means of DNA analysis with closely linked polymorphic markers and the definite diagnosis based on sequencing data were found.


Relation of the ventromedial nuclei of the hypothalamus to the regulation of renin secretion.
During an investigation of the role of the mediobasal hypothalamus in the regulation of renin secretion from the kidneys, we found that lesions of the ventromedial nuclei prevented the increase in plasma renin activity produced by p-chloroamphetamine. In the present study, we tested the effects of bilateral electrolytic lesions of the ventromedial nuclei on the increase in plasma renin activity produced in sham-operated rats by immobilization, head-up tilt under inactin anesthesia, and a low-sodium diet. Ventromedial lesions reduced or abolished the plasma renin activity increase to all three stimuli without any change in plasma angiotensinogen. The plasma renin concentration responses to immobilization and a low-sodium diet were also reduced. All these stimuli probably exert their effect by way of the sympathetic nervous system. The data support the hypothesis that the ventromedial nuclei or neural fibers passing through them are important in the renin response to diverse stimuli that act by way of sympathetic discharge.


Discovery of TDI-10229: A Potent and Orally Bioavailable Inhibitor of Soluble Adenylyl Cyclase (sAC, ADCY10).
Soluble adenylyl cyclase (sAC) has gained attention as a potential therapeutic target given the role of this enzyme in intracellular signaling. We describe successful efforts to design improved sAC inhibitors amenable for in vivo interrogation of sAC inhibition to assess its potential therapeutic applications. This work culminated in the identification of TDI-10229 (12), which displays nanomolar inhibition of sAC in both biochemical and cellular assays and exhibits mouse pharmacokinetic properties sufficient to warrant its use as an in vivo tool compound.


Experiments on Temperature Changes of Microbolometer under Blackbody Radiation and Predictions Using Thermal Modeling by COMSOL Multiphysics Simulator.
In this study, we study a heat transfer model, with the surface of the microbolometer device receiving radiation from blackbody constructed using a COMSOL Multiphysics simulator. We have proposed three kinds of L-type 2-leg and 4-leg with the pixel pitch of 35 μm based on vanadium oxide absorbent membrane sandwiched with top passivated and bottom Si₃N₄ supporting films, respectively. Under the blackbody radiation, the surface temperature changes and distributions of these samples are simulated and analyzed in detail. The trend of change of the temperature dependent resistance of the four kinds of bolometer devices using the proposed heat transfer model is consistent with the actual results of the change of resistance of 4 samples irradiated with 325 K blackbody located in the front distance of 5 cm. In this paper, ΔT indicates the averaged differences of the top temperature on the suspended membrane and the lowest temperature on the post of legs of the microbolometers. It is shown that ΔT ≈ 17 mK is larger in nominal 2-leg microbolometer device than that of 4-leg one and of 2-leg with 2 μm × 2 μm central square hole and two 7.5 μm × 2 μm slits in suspended films. Additionally, only ΔT ≈ 5 mK with 4-leg microbolometer device under the same radiated energy of 325 K blackbody results from the larger total thermal conductance.


A Cluster-Randomized Evaluation of the SuperShelf Intervention in Choice-Based Food Pantries.
Interventions in food pantry settings have the potential to improve health among clients at risk of diet-related disease. This study evaluates whether a cluster-randomized, behavioral intervention in food pantries resulted in improved client outcomes. Sixteen Minnesota food pantries were randomized to an intervention (n = 8) or control condition (n = 8). The intervention offered pantries technical assistance to improve healthy food supply and implement behavioral economics strategies to promote healthy food selection. A convenience sample of adult clients were enrolled (paired sample, 158 intervention, 159 control) and followed for 1 year. Additional clients were enrolled at follow-up to assess food selection (follow-up sample, 85 intervention, 102 control). Analysis was limited to data from 11 pantries (5 intervention, 6 control) due to COVID-19. Outcome measures included Healthy Eating Index-2015 (HEI-2015) total and subcomponent scores for 24-hr dietary recalls and client cart selections, and Life's Simple 7 (LS7) total and subcomponent scores. Multilevel mixed-effects models tested whether client outcomes differed by intervention condition. In adjusted models, there were no statistically significant differences by intervention condition in HEI-2015 or LS7 scores. Clients in intervention food pantries had improved Refined Grain subcomponent scores (p = .004); clients in control pantries had worsened Saturated Fat subcomponents scores (p = .019) and improved physical activity scores (p = .007). The intervention did not result in improved diet quality or cardiovascular health as measured by HEI-2015 or LS7. Coordinated efforts across settings are needed to address health risks facing this population.


Active-imaginal exposure: examination of a new behavioral treatment for cynophobia (dog phobia).
The aims of this study were to investigate exposure-based treatments for cynophobia (dog phobia) and to test a newly developed hybrid imaginal exposure treatment that we have named active imaginal exposure. The treatment introduces an in vivo coping component to imaginal exposure whereby the patient physically performs coping responses to an imagined feared stimulus. Eighty-two participants meeting DSM-IV criteria for specific phobia (animal subtype) were randomly assigned to one of three 30-min. treatments: (a) active-imaginal exposure (AI), (b) imaginal exposure alone (IE), or (c) graduated in vivo exposure (IV). Participants completed a behavioral approach test at pre, post, and four-week follow-up. Significant pre- to posttreatment improvement was observed in all three treatment conditions. Response rates at posttreatment were 51.9, 62.1, and 73.1% for the IE, AI, and IV groups respectively. Likewise, effect sizes at posttreatment were 0.76, 1.41, and 1.55 for the IE, AI, and IV groups respectively. Although in the predicted direction, the between group differences were not significant. A similar pattern of results was observed at follow-up. Further, safety behavior utilization during treatment was associated with less improvement--particularly in the two imaginal treatment conditions. Exposure treatments of dog phobia appear feasible and effective in reducing phobic fear and avoidance associated with dog phobia. Furthermore, preliminary evidence suggests that our active-imaginal exposure treatment may be a viable alternative to in vivo exposure.


A bioinspired route to various siliceous vesicular structures.
Various siliceous nanostructures have been successfully synthesized through the co-organization of organic molecules and inorganic silica source under mild pH conditions (pH approximately 5). A biodegradable block copolymer P123 [EO20PO70EO20, EO is poly (ethylene oxide), PO is poly (propylene oxide)] is employed as a marcomolecular template and Na2SiO39H2O as a silica source. By changing the concentrations of the reactants and/or reaction temperature, siliceous multilamellar vesicles, unilamellar nano-foams and multilamellar vesicles with sponge-like walls have been obtained. Our work provides a convenient and bioinspired route to obtain siliceous nanostructured materials with adjustable and multi-level pore structures as well as rich morphologies, which is important to understand the biomineralization mechanism. Such artificial silica nanoporous materials may find potential applications in catalysis, separations, electronics, and photonics, etc.


Disseminated Mycobacterium avium subsp. avium infection in a pet Korean squirrel (Sciuris vulgaris coreae).
A disseminated Mycobacterium avium subsp. avium infection was diagnosed in a pet Korean squirrel. Grossly, multiple small nodules in the lung, liver, spleen, and skin were observed. Adrenal glands were very enlarged. The only tissue exhibiting necrosis and calcification was a very enlarged bronchial lymph node. The remaining lymph nodes were slightly enlarged. Moderate ascites was also observed. Microscopically, a disseminated granulomatous inflammation with numerous lymphocytes was seen. Acid-fast bacilli were detected in macrophages, in giant cells, free in the interstitium, and in some lymphatic vessels, both within cells and free in the lumen. M. avium subsp. avium was isolated and identified by polymerase chain reaction-restriction endonuclease analysis.


The potential anti-cancer effects of quercetin on blood, prostate and lung cancers: An update.
Cancer is caused by abnormal proliferation of cells and aberrant recognition of the immune system. According to recent studies, natural products are most likely to be effective at preventing cancer without causing any noticeable complications. Among the bioactive flavonoids found in fruits and vegetables, quercetin is known for its anti-inflammatory, antioxidant, and anticancer properties. This review aims to highlight the potential therapeutic effects of quercetin on some different types of cancers including blood, lung and prostate cancers.


Does nefazodone improve both depression and Parkinson disease? A pilot randomized trial.
Some of the selective serotonin reuptake inhibitors (SSRI)-induced motor side effects are mediated by stimulating 5-HT2 receptors in the basal ganglia, probably because serotonin inhibits the subsequent neuronal dopamine release. We hypothesized that nefazodone, a serotonin 2 antagonist/reuptake inhibitor (SARI) that selectively blocks 5-HT2 receptors, could disrupt the aforementioned inhibitory pathway. Therefore, increased dopamine levels in the postsynaptic milieu and an improvement in the motor symptoms in depressed patients with Parkinson disease (PD) should be observed. This study was designed to determine whether nefazodone has a dual activity as an antidepressant and as an agent capable of reducing the extrapyramidal symptoms in depressed parkinsonian patients. Depressed patients with PD were randomly assigned to 2 therapeutic groups: nefazodone or fluoxetine. Patients were evaluated by a psychiatrist and were blindly assessed by a neurologist with an array of scales. Patients on nefazodone (n = 9) showed a significant improvement over time in the total Unified Parkinson Disease Rating Scale score (UPDRS) (part II + part III) (P = 0.004) and in the UPDRS subscore part III (P = 0.003). None of these scores changed over time in the fluoxetine group (n = 7). Both, nefazodone and fluoxetine were equally effective as antidepressants: Beck Depression Inventory scores significantly improved (P < 0.001), with no significant differences between treatment groups (P = 0.97). If our results can be confirmed in a larger clinical trial, nefazodone ought to be considered over fluoxetine given its secondary beneficial effects regarding the reduction of extrapyramidal symptoms in depressed PD patients.


A Neural Dynamic Architecture for Reaching and Grasping Integrates Perception and Movement Generation and Enables On-Line Updating.
Reaching for objects and grasping them is a fundamental skill for any autonomous robot that interacts with its environment. Although this skill seems trivial to adults, who effortlessly pick up even objects they have never seen before, it is hard for other animals, for human infants, and for most autonomous robots. Any time during movement preparation and execution, human reaching movement are updated if the visual scene changes (with a delay of about 100 ms). The capability for online updating highlights how tightly perception, movement planning, and movement generation are integrated in humans. Here, we report on an effort to reproduce this tight integration in a neural dynamic process model of reaching and grasping that covers the complete path from visual perception to movement generation within a unified modeling framework, Dynamic Field Theory. All requisite processes are realized as time-continuous dynamical systems that model the evolution in time of neural population activation. Population level neural processes bring about the attentional selection of objects, the estimation of object shape and pose, and the mapping of pose parameters to suitable movement parameters. Once a target object has been selected, its pose parameters couple into the neural dynamics of movement generation so that changes of pose are propagated through the architecture to update the performed movement online. Implementing the neural architecture on an anthropomorphic robot arm equipped with a Kinect sensor, we evaluate the model by grasping wooden objects. Their size, shape, and pose are estimated from a neural model of scene perception that is based on feature fields. The sequential organization of a reach and grasp act emerges from a sequence of dynamic instabilities within a neural dynamics of behavioral organization, that effectively switches the neural controllers from one phase of the action to the next. Trajectory formation itself is driven by a dynamical systems version of the potential field approach. We highlight the emergent capacity for online updating by showing that a shift or rotation of the object during the reaching phase leads to the online adaptation of the movement plan and successful completion of the grasp.


Site of the arrhythmogenic focus and cardiac vulnerability to ventricular fibrillation.
The aim of this study was to test the hypothesis that a subendocardial arrhythmogenic focus makes the heart more susceptible to VF due to electrical interaction with the Purkinje network. Monofocal ventricular tachycardia (mVT) was created by injecting 5-microg aconitine into the left ventricular subepicardium (EPI-mVT, n = 8) or subendocardium (ENDO-mVT, n = 13) in anesthetized dogs. Despite the similar cycle length of mVT, the incidence of VF was significantly different between EPI-mVT and ENDO-mVT (100 [8/8] vs 46% [6/13], P <0.05). VF was invariably preceded by hemodynamic deterioration. Three-dimensional cardiac mapping (n = 10, 221 +/- 11 recording sites) revealed that VF was triggered solely by focal firing unrelated to the primary arrhythmogenic focus in both mVT models. No interaction between the primary focus and adjacent endocardial tissue was indicated. These results suggest that the proximity of the arrhythmogenic focus to the Purkinje network has little role in cardiac vulnerability to VF, and that progression of mVT to VF is largely caused by sporadic focal firing regardless of the site of the arrhythmogenic focus in the present animal model.


The Commit to Be Fit framework: a community case study of a multi-level, holistic school-based wellness initiative in rural Virginia.
Public health interventions that target children's physical, mental, and emotional health will enhance their ability to learn and grow. Although more complex, school initiatives that address multiple ecological levels and take a holistic view may be more effective and likely to lead to lasting change. This article presents the framework of Commit to Be Fit (C2BF) as an example of how schools can integrate multi-level and holistic approaches for health. This innovative school-based intervention includes activities addressing individual, home, school, and community to create a culture of wellness. We describe the implementation of C2BF and its basis in ecological models and give examples of activities across three components: cafeteria, classroom, and community. We discuss challenges and note that leadership engagement and alignment were critical elements for C2BF's success thus far. C2BF uses a school-based multi-level approach to creating a culture of wellness and holistic health for students, teachers, and community members. C2BF is unique compared to other school-based programming and includes activities that address all eight domains posited for program sustainability within public health. Built to be flexible and adaptive, C2BF was able to successfully pivot during the COVID pandemic and also follow new science. C2BF and other multi-level holistic approaches are more likely to achieve long-term change by utilizing strategies across the multiple levels of the ecological model to improve health and wellbeing.


Flavor characterization of top-blade, top-sirloin, and tenderloin steaks as affected by pH, maturity, and marbling.
Little information is available in the literature on the interrelationships and interactions among pH, aging time, marbling, and maturity on the flavor profile of some beef muscles commonly used for steaks. To investigate these effects on beef flavor, the infraspinatus (top-blade steak) from the chuck clod, the gluteus medius (top-sirloin steak) from the sirloin, and the psoas major (tenderloin steak) from the loin were obtained from A- (n = 80) and B-maturity (n = 60) carcasses with either Slight (n = 68) or Small (n = 72) marbling, and with either normal (< or = 5.7; n = 80) or high (> or = 6.0; n = 60) pH. Muscles were selected from two commercial processing plants at six different sampling times to evaluate factors that affect the flavor profile of cooked beef steaks. Muscles were vacuum-aged for 7, 14, 21, or 35 d, and a highly trained, flavor-profile sensory panel evaluated charbroiled steaks from these muscles. Numerous statistical interactions (P < 0.05) were detected for flavor attributes of the different muscles. In general, muscles from high pH (dark cutting) carcasses had less typical beef flavor identity and less brown-roasted flavor than those from carcasses with normal pH. Aging longer than 21 d generally decreased beef flavor identity. Top-blade steaks generally had less intense beef flavor identity and more intense bloody/serumy flavor than did top-sirloin and tenderloin steaks. Tenderloin and top-sirloin steaks of normal pH generally had the most brown-roasted flavor, especially when aged 21 d or less. Small degree of marbling generally resulted in a more rancid flavor compared with Slight marbling, but marbling had no other appreciable effects on the flavor profile. Aging steaks for 35 d increased (P < 0.05) the metallic flavor compared with aging for only 7 or 14 d. Top-sirloin steaks had a more intense (P < 0.05) sour flavor than did top-blade steaks, and steaks from carcasses with a high pH were more rancid (P < 0.05) than steaks from carcasses with normal pH. Vacuum-aging top-blade, top-sirloin, and tenderloin steaks to 21 or 35 d postmortem generally increased metallic and rancid flavors and increased sour flavor in top-sirloin steaks that were high in pH.


Metabolic and synthetic activities of Pneumocystis carinii in vitro.
Pneumocystic carinii organisms obtained from the lungs of rats with glucocorticoid-induced pneumocystosis retained a number of metabolic activities during in vitro culture for 48 to 72 h. They were able to metabolize [14C]glucose to 14CO2, to synthesize proteins from 3H-amino acids, and to synthesize ribonucleic acid from [3H]uridine. Utilization of [14C]glucose was inhibitable by excess unlabeled glucose, mannose, fructose, 2-deoxy-D-glucose, iodoacetate, and pentamidine, but not glycerol or trimethoprim-sulfamethoxazole. Protein synthesis was almost completely inhibited by incubation with high concentrations of cycloheximide, pentamidine, suramin, and chloroquine, but not by rifampin, tetracycline, or gentamicin. In additional experiments, P. carinii organisms that had been incubated overnight in the presence of potentially lethal agents were tested for their ability to utilize [14C]glucose in inhibitor-free medium as a test of viability. Pentamidine, at concentrations which caused only modest inhibition of glucose metabolism, protein synthesis, amino acid transport, or ribonucleic acid synthesis, was lethal, whereas trimethoprim-sulfamethoxazole was not.


[Pathophysiology of primary testicular dysfunction].
This review summarizes the most important forms of primary testicular dysfunction with particular reference to new pathophysiological concepts that may lead to a better understanding of primary testicular dysfunctions. The observed abnormalities can be used as a model to understand the multiple interactions of the different gonadal compartments.


Study of mercury content in edible mushrooms.
The influence of time and temperature of mineralisation of mushrooms was studied for mercury determination by cold vapor atomic absorption. Optimal conditions for a complete mercury recuperation were obtained using a mixture of nitric and sulphuric acid (6 ml HNO3, 4 ml H2SO4) in presence of KMnO4 (3 ml of a 0.44 M solution) at 95 degrees C for 4 h, with 1 g wet sample. The detection limit of method was 3.2 ng/g. These conditions have shown high reproductibility and accuracy.


Rod-to-Globule Transition of pDNA/PEG-Poly(l-Lysine) Polyplex Micelles Induced by a Collapsed Balance Between DNA Rigidity and PEG Crowdedness.
The role of poly(ethylene-glycol) (PEG) in rod-shaped polyplex micelle structures, having a characteristic core of folded plasmid DNA (pDNA) and a shell of tethered PEG chains, is investigated using PEG-detachable polyplex micelles. Rod shapes undergo change to compacted globule shapes by removal of PEG from polyplex micelles prepared from block copolymer with acid-labile linkage between PEG and poly(l-lysine) (PLys) through exposure to acidic milieu. This structural change supports the previous investigation on the rod shapes that PEG shell prevents the DNA structure from being globule shaped as the most favored structure in minimizing surface area. Noteworthy, despite the PEG is continuously depleted, the structural change does not occur in gradual shortening manner but the rod shapes keep their length unchanged and abruptly transform into globule shapes. Analysis of PEG density reveals the transition occurred when tethered PEG of rod shapes has decreased to a critical crowdedness, i.e., discontacted with neighboring PEG, which eventually illuminates another contribution, rigidity of DNA packaged as bundle in the rod shapes, in addition to the steric repulsion of PEG, in sustaining rod shapes. This investigation affirms significant role of PEG and also DNA rigidity as bundle in the formation of rod-shaped structures enduring the quest of compaction of charge-neutralized DNA in the polyplex micelles.


Water compartments in cells.
Human experience in the macrobiological world leads scientists to visualize water compartments in cells analogous to the bladder in the human pelvis or ventricles in the brain. While such water-filled cellular compartments likely exist, the volume contributions are insignificant relative to those of biomolecular hydration compartments. The purpose of this chapter is to identify and categorize the molecular water compartments caused by proteins, the primary macromolecular components of cells. The categorical changes in free energy of water molecules on proteins cause these compartments to play dominant roles in osmoregulation and provide important adjuncts to fundamental understanding of osmosensing and osmosignaling mechanisms. Water compartments possess differences in molecular motion, enthalpy, entropy, freezing point depression, and other properties because of electrostatic interaction of polar water molecules with electric fields generated by covalently bound pairs of opposite charge caused by electronegative oxygen and nitrogen atoms of the protein. Macromolecules, including polypeptides, polynucleotides, and polysaccharides, are stiff molecular chains with restricted folding capacities due to inclusion of rigid ring structures or double amide bonds in the backbone sequence. This creates "irreducible spatial charge separation" between positive and negative partial charges, causing elevated electrostatic energy. In the fully hydrated in vivo state of living cells the high dielectric coefficient of water reduces protein electrostatic free energy by providing polar "water bridge networks" between charges, thereby creating four measurably different compartments of bound water with distinct free energy differences.


Voltammetric analysis for fast and inexpensive diagnosis of urinary tract infection: a diagnostic study.
Dipstick test is widely used to support the diagnosis of urinary tract infections (UTI). It is effective in ruling out UTI, but urine culture is needed for diagnosis confirmation. In this study we compared the accuracy of voltammetric analysis (VA) with that of DT to detect UTI (diagnosed using urine culture), and its usefulness as a second-stage test in people with positive DT. 142 patients were enrolled with no exclusion criteria. VA was performed using the BIONOTE device. Partial Least Square Discrimination Analysis was used to predict UTI based on VA data; diagnostic performance was evaluated using sensitivity, specificity, positive and negative predictive values (PPV and NPV, respectively), positive and negative likelihood ratios (LR), accuracy, diagnostic odds ratio (DOR). Mean age was 76.6 years (SD 12.6), 57% were male. VA had a better overall performance respect to DT in detecting UTI with accuracy 81.7% vs 75.9%, specificity 90.8% vs 82.5%, PPV 75% vs 61.4%, positive LR 6.68 vs 3.5, DOR 17.7 vs 7.47; sensibility, NPV and negative LR of the two tests were similar. VA had an accuracy of 82.4% in discriminating bacterial from fungal infections. When added as a second-stage test, VA identified 9 of the 17 false positive patients, with a net specificity of 91.7%, sensitivity 54%, PPV 75% and NPV 81%. VA is a quick and easy method that may be used as a second stage after DT to reduce the number of urine culture and of inappropriate antibiotic prescriptions.


Synthesis of Zwitterionic and Trehalose Polymers with Variable Degradation Rates and Stabilization of Insulin.
Polymers that stabilize biomolecules are important as excipients in protein formulation. Herein, we describe a class of degradable polymers that have tunable degradation rates depending on the polymer backbone and can stabilize proteins to aggregation. Specifically, zwitterion- and trehalose-substituted polycaprolactone, polyvalerolactone, polycarbonate, and polylactide were prepared and characterized with regards to their hydrolytic degradation and ability to stabilize insulin to mechanical agitation during heat. Ring-opening polymerization (ROP) of allyl-substituted monomers was performed by using organocatalysis, resulting in well-defined alkene-substituted polymers with good control over molecular weight and dispersity. The polymers were then modified by using photocatalyzed thiol-ene reactions to install protein-stabilizing carboxybetaine and trehalose side chains. The resulting polymers were water-soluble and exhibited a wide range of half-lives, from 12 h to more than 3 months. The polymers maintained the ability to stabilize the therapeutic protein insulin from activity loss due to aggregation, demonstrating their potential as degradable excipients for protein formulation.


An in vitro study of the susceptibility of mobile and cystic forms of Borrelia burgdorferi to tinidazole.
The susceptibility of mobile and cystic forms of Borrelia burgdorferi to tinidazole (TZ) was examined. The minimal bactericidal concentration (MBC) of TZ against the mobile spirochetes was >128 microg/ml at 37 degrees C in micro-oxic atmosphere when incubated for 14 days. TZ significantly reduced the conversion of mobile spirochetes to cystic forms during incubation. The MBC for older (10-months-old) cysts at 37 degrees C in a micro-oxic atmosphere was >0.5 microg/ml, but >0.125 microg/ml for young (1-day-old) cysts. Acridine orange staining, dark-field microscopy and transmission electron microscopy revealed that, when the concentration of TZ was > or = MBC, the contents of the cysts were partly degraded, core structures did not develop inside the young cysts, and the amount of RNA in these cysts decreased significantly. When cysts were exposed to TZ, both the spirochetal structures and core structures inside the cysts dissolved, and the production of blebs was significantly reduced. These observations may be valuable in the treatment of resistant infections caused by B. burgdorferi, and suggest that a combination of TZ and a macrolide antibiotic could eradicate both cystic and mobile forms of B. burgdorferi.


A Technical Approach to the Evaluation of Radiofrequency Radiation Emissions from Mobile Telephony Base Stations.
During the last two decades, the number of macrocell mobile telephony base station antennas emitting radiofrequency (RF) electromagnetic radiation (EMR) in residential areas has increased significantly, and therefore much more attention is being paid to RF EMR and its effects on human health. Scientific field measurements of public exposure to RF EMR (specifically to radio frequency radiation) from macrocell mobile telephony base station antennas and RF electromagnetic field (EMF) intensity parameters in the environment are discussed in this article. The research methodology is applied according to the requirements of safety norms and Lithuanian Standards in English (LST EN). The article presents and analyses RF EMFs generated by mobile telephony base station antennas in areas accessible to the general public. Measurements of the RF electric field strength and RF EMF power density were conducted in the near- and far-fields of the mobile telephony base station antenna. Broadband and frequency-selective measurements were performed outside (on the roof and on the ground) and in a residential area. The tests performed on the roof in front of the mobile telephony base station antennas in the near-field revealed the presence of a dynamic energy interaction within the antenna electric field, which changes rapidly with distance. The RF EMF power density values on the ground at distances of 50, 100, 200, 300, 400, and 500 m from the base station are very low and are scattered within intervals of 0.002 to 0.05 μW/cm². The results were compared with international exposure guidelines (ICNIRP).


NO sensitizes rat hepatocytes to proliferation by modifying S-adenosylmethionine levels.
Liver regeneration is a fundamental response of this organ to injury. Hepatocyte proliferation is triggered by growth factors, such as hepatocyte growth factor. However, hepatocytes need to be primed to react to mitogenic signals. It is known that nitrous oxide (NO), generated after partial hepatectomy, plays an important role in hepatocyte growth. Nevertheless, the molecular mechanisms behind this priming event are not completely known. S-adenosylmethionine (AdoMet) synthesis by methionine adenosyltransferase is the first step in methionine metabolism, and NO regulates hepatocyte S-adenosylmethionine levels through specific inhibition of this enzyme. We have studied the modulation of hepatocyte growth factor-induced proliferation by NO through the regulation of S-adenosylmethionine levels. Studies were conducted in cultured rat hepatocytes isolated by collagenase perfusion, which triggers NO synthesis. The mitogenic response to hepatocyte growth factor was blunted when inducible NO synthase was inhibited; this process was overcome by the addition of an NO donor. This effect was dependent on methionine concentration in culture medium and intracellular S-adenosylmethionine levels. Accordingly, we found that S-adenosylmethionine inhibits hepatocyte growth factor-induced cyclin D1 and D2 expression, activator protein 1 induction, and hepatocyte proliferation. Together our findings indicate that NO may switch hepatocytes into a hepatocyte growth factor-responsive state through the down-regulation of S-adenosylmethionine levels.


Leaders, self-confidence, and hubris: what's the difference?
Success can easily breed hubris. As leaders become more confident, their success can limit their learning because they develop repetitive patterns of filtering information based on past successes and discount information that does not agree with their patterns of success. It is important for leaders to stay grounded in reality and effective as their success grows. Humility, gratitude, and appreciation will avoid the overconfidence that leads to hubris. Building confidence in others is the mark of a great leader. Hubris is not.


Heter-LP: A heterogeneous label propagation algorithm and its application in drug repositioning.
Drug repositioning offers an effective solution to drug discovery, saving both time and resources by finding new indications for existing drugs. Typically, a drug takes effect via its protein targets in the cell. As a result, it is necessary for drug development studies to conduct an investigation into the interrelationships of drugs, protein targets, and diseases. Although previous studies have made a strong case for the effectiveness of integrative network-based methods for predicting these interrelationships, little progress has been achieved in this regard within drug repositioning research. Moreover, the interactions of new drugs and targets (lacking any known targets and drugs, respectively) cannot be accurately predicted by most established methods. In this paper, we propose a novel semi-supervised heterogeneous label propagation algorithm named Heter-LP, which applies both local and global network features for data integration. To predict drug-target, disease-target, and drug-disease associations, we use information about drugs, diseases, and targets as collected from multiple sources at different levels. Our algorithm integrates these various types of data into a heterogeneous network and implements a label propagation algorithm to find new interactions. Statistical analyses of 10-fold cross-validation results and experimental analyses support the effectiveness of the proposed algorithm.


Fast chemical reaction in two-dimensional Navier-Stokes flow: initial regime.
This paper studies an infinitely fast bimolecular chemical reaction in a two-dimensional biperiodic Navier-Stokes flow. The reactants in stoichiometric quantities are initially segregated by infinite gradients. The focus is placed on the initial stage of the reaction characterized by a well-defined one-dimensional material contact line between the reactants. Particular attention is given to the effect of the diffusion κ of the reactants. This study is an idealized framework for isentropic mixing in the lower stratosphere and is motivated by the need to better understand the effect of resolution on stratospheric chemistry in climate-chemistry models. Adopting a Lagrangian straining theory approach, we relate theoretically the ensemble mean of the length of the contact line, of the gradients along it, and of the modulus of the time derivative of the space-average reactant concentrations (here called the chemical speed) to the joint probability density function of the finite-time Lyapunov exponent λ with two times τ and τ[over ̃]. The time 1/λ measures the stretching time scale of a Lagrangian parcel on a chaotic orbit up to a finite time t, while τ measures it in the recent past before t, and τ[over ̃] in the early part of the trajectory. We show that the chemical speed scales like κ(1/2) and that its time evolution is determined by rare large events in the finite-time Lyapunov exponent distribution. The case of smooth initial gradients is also discussed. The theoretical results are tested with an ensemble of direct numerical simulations (DNSs) using a pseudospectral model.


Natural killer cellular cytotoxicity against herpes simplex virus-infected cells in Igh-1-disparate mice.
Susceptibility to Herpes simplex virus type 1 (HSV-1) stromal keratitis (HSK) in the mouse has previously been linked to the Igh-1 locus. The role of natural killer cells (NK) in resistance to viral infections is controversial. The authors studied the influence of the Igh-1 locus on in vitro murine NK activity against HSV-1 infected cell lines. The HSV-1 infected targets were lysed better than uninfected cells by murine splenic lymphocytes. Strain had no influence on virus-augmented cell lysis. Spleen cells from naive HSK-susceptible CAL-20 (Igh-1d) and BALB/c (Igh-1a) mice lysed YAC-1 targets better than HSK-resistant C.B-17 (Igh-1b) mice. The reverse was seen 24 hours after in vivo infection intraperitoneally with HSV-1. In contrast, CAL-20 splenocytes lysed PU5-1R targets better than BALB/c and C.B-17 splenocytes 24 hours after intraperitoneal (IP) infection. No significant differences were detected in interferon (IFN) levels after IP challenge with HSV-1 among the Igh-1 congenics. The data show that differences in NK activity were determined by both the Igh-1 genotype and the uninfected target cell. Susceptibility to HSK in these Igh-1-disparate congenics thus cannot be explained simply by differences in NK activity against HSV-1-infected targets.


New-Onset Dementia Among Survivors of Pneumonia Associated With Severe Acute Respiratory Syndrome Coronavirus 2 Infection.
Case series without control groups suggest that severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection may result in cognitive deficits and dementia in the postinfectious period. Adult pneumonia patients with SARS-CoV-2 infection (index hospitalization) and age-, gender-, and race/ethnicity-matched contemporary control pneumonia patients without SARS-CoV-2 infection were identified from 110 healthcare facilities in United States. The risk of new diagnosis of dementia following >30 days after the index hospitalization event without any previous history of dementia was identified using logistic regression analysis to adjust for potential confounders. Among 10 403 patients with pneumonia associated with SARS-CoV-2 infection, 312 patients (3% [95% confidence interval {CI}, 2.7%-3.4%]) developed new-onset dementia over a median period of 182 days (quartile 1 = 113 days, quartile 3 = 277 days). After adjustment for age, gender, race/ethnicity, hypertension, diabetes mellitus, hyperlipidemia, nicotine dependence/tobacco use, alcohol use/abuse, atrial fibrillation, previous stroke, and congestive heart failure, the risk of new-onset dementia was significantly higher with pneumonia associated with SARS-CoV-2 infection compared with pneumonia unrelated to SARS-CoV-2 infection (odds ratio [OR], 1.3 [95% CI, 1.1-1.5]). The association remained significant after further adjustment for occurrence of stroke, septic shock, and intubation/mechanical ventilation during index hospitalization (OR, 1.3 [95% CI, 1.1-1.5]). Approximately 3% of patients with pneumonia associated with SARS-CoV-2 infection developed new-onset dementia, which was significantly higher than the rate seen with other pneumonias.


Curricular Approaches to Transgender Health in Physician Assistant Education.
According to the Williams Institute, 1.4 million U.S. adults identify as transgender. Many experience health care disparities. Professional organizations call for medical education to improve transgender care, but what curricula are being delivered is unknown. The goal of this study was to conduct the first comprehensive, national survey of transgender health care curricula in physician assistant (PA) education. The authors sent a questionnaire to program directors (PDs) at all 236 U.S. PA programs in June 2018. They categorized programs as those that currently deliver at least 1 hour of transgender health content and those who do not (Teaching/Not Teaching). They examined differences between Teaching and Not Teaching programs using chi-square tests, and they evaluated comments for themes. The response rate was 100%. Of the 236 programs, 202 (85.6%) teach transgender content and 34 (14.4%) do not. According to PDs, most transgender content was delivered in medical interviewing (44.1%) or women's health (31.4%) and the most common transgender health topics included differentiating between sex and gender and between behavior and identity (78.8%), followed by health disparities (77.5%) and barriers to care (75.0%). PDs most commonly cited a lack of time (51.3%) and faculty knowledge (35.6%) as barriers for teaching transgender health topics. Half of the PDs (50.4%) ranked transgender health as very or extremely important. The authors detected statistically significant differences between Teaching and Not Teaching programs based on geographic region (P = .01), perceived importance (P ≤ .001), and presence of knowledgeable faculty (P = .01). Presence of knowledgeable faculty was significantly associated with perceived importance (P = .01). This is the first comprehensive, nationwide survey of transgender health education in U.S. PA programs. A key finding is that the presence of expert faculty is significantly associated with delivery and perceived importance of transgender health curricula.


A rapid in vitro electrochemical screening of extracellular matrix of Saccharomyces cerevisiae by palladium nanoparticles-modified electrodes.
Herein, a rapid electrochemical screening of yeasts (Saccharomyces cerevisiae) in vitro mode depending on their optical density, cultivation time and growth medium used was conducted in 3 min by palladium nanoparticles (Pd-NPs)-modified electrodes. Pd-NPs-modified electrodes operated in cyclic voltammetry mode at low scan rates, i.e. 5-20 mV/s supported a low oxidative process in the yeast extracellular matrix. The electrochemical screening relied on an efficient electrooxidation of secondary metabolites, i.e. organohydrazines formed in the extracellular medium as a result of microbial activity of yeast cells. More importantly, during the study the impact of fundamental parameters, viz. type of the matrix and pH on electroanalytical response of Pd-NPs-based electrodes in real fermentation medium was investigated in detail. The efficiency of the proposed in vitro electrochemical screening of yeast extracellular matrix was not affected by pH of the samples or composition of the multicomponent medium, but more likely exclusively depended on the presence of organohydrazines. The potential of this electroanalytical approach towards profiling of the extracellular matrix of Saccharomyces cerevisiae was compared with results obtained by gas chromatography mass-spectrometry (GC-MS) and genetically encoded biosensor (ro-GFP2) assays.


Preoperative IL-2 immunotherapy enhances tumor infiltrating lymphocytes (TILs) in gastric cancer patients.
Tumor infiltrating lymphocytes (TILs), recognized as a tumor-host reaction, have been linked to prognosis in various tumors, with a clear positive correlation between the density of the lymphoid infiltrate at the advancing margin of the tumor and the prognosis of the patients. TILs are somewhat activated by tumor associated antigens and by IL-2 endogenous release. The aim of this study is to verify if subcutaneously administered IL-2 is able to enhance TILs in gastric cancer patients and is able to influence the prognosis of the patients. We enrolled 39 consecutive patients with gastric adenocarcinoma. Patients were randomized to be treated with surgery alone (control group, 20 patients) or with surgery plus preoperative IL-2 (Interleukin-2) administration (treated group, 19 patients). Total lymphocytes, CD4 and CD4/CD8 were evaluated pre- and postoperatively. Peritumoral stromal reaction, neutrophils, lymphocytes and eosinophils infiltration in tumor histology were evaluated as well as survival curves and compared between the groups. IL-2 treatment was safe and well tolerated, and in the IL-2 treated group a significant increase over the baseline pretreatment values of the total lymphocyte, CD4 and CD4/CD8 on both the 14th and 50th postoperative days was observed (p < 0.05). Peritumoral stromal reaction, neutrophils and eosinophils infiltration did not shown any statistical difference between the two groups. Otherwise we observed a statistically significant difference in the peri- and intratumoral lymphocytes infiltration between IL-2 treated and control patients (p = 0.000026). Median overall and disease-free survivals were longer, even if not significantly, in the IL-2 group than in the control arm (p = 0.089 and p = 0.09 respectively). Our data shows that IL-2 seems to be able to induce substantial changes in the inflammatory infiltration of the neoplasm, improving the host activity toward the tumor and enhancing the TILs phenomenon in gastric cancer patients. This feature seems to improve the prognosis of the patients.


Applying the stages-of-change model to dietary change.
The transtheoretical model of behavior change ("Stages of Change") was first proposed by Prochaska and DiClemente 14 years ago. It was originally developed by observing smokers who were planning or attempting to give up smoking. It has since been applied to behaviors other than smoking, and several recently published papers have examined its application in the area of dietary change. The complexity of dietary change, however, has made it more difficult to apply the model in this area. Studies applying the model to diet have differed in terms of the aspect of diet being examined, as well as the staging algorithms and dietary assessment methodology used. Such differences in methodology have led to variable results and have made it difficult to interpret results obtained. This review summarizes the studies in this area and makes recommendations for future research.


Orally administered FTS (salirasib) inhibits human pancreatic tumor growth in nude mice.
S-trans,trans-farnesylthiosalicylic acid (salirasib, FTS) is a synthetic small molecule that acts as a potent Ras inhibitor. Salirasib inhibits specifically both oncogenically activated Ras and growth factor receptor-mediated Ras activation, resulting in the inhibition of Ras-dependent tumor growth. The objectives of this study were to develop a sensitive LC-MS/MS assay for determination of FTS in plasma, to assess the bioavailabilty of FTS after oral administration to mice, and then to examine the efficacy of orally administered FTS for inhibition of tumor growth in a nude mouse model. FTS was isolated from mouse plasma by liquid chromatography on a Columbus 5-mum particle size, 50 x 2 mm id column with a methanol/5 mM ammonium acetate (80/20) mobile phase (isocratic elution) at a flow rate of 0.3 ml/min. MS/MS was performed on a PE Sciex API 365 with Turbo Ion Spray as interface and negative ion ionization; parent ion (m/z): 357.2; daughter ion (m/z) 153.2; retention time 2.3 min. For plasma analysis, the amount of analyte in each sample was calculated by comparing response of the analyte in that sample to a nine-point standard curve linear over the range 3-1000 ng/ml. Pharmacokinetic studies were performed in mice following intraperitoneal dosing (20 mk/kg in PBS) or oral dosing (40 mg/kg in either 0.5% aqueous CMC or corn oil). Panc-1 tumor growth in nude mice was determined following daily oral dosing with FTS in 0.5% CMC (40, 60, or 80 mg/kg), or in combination with weekly gemcitabine (30 mg/kg). Salirasib was readily detected in mouse plasma by LC-MS/MS at a detection limit of 3 ng/ml. For each route of administration, t (max) was 1 h and t (1/2) ranged from 1.86 to 2.66 h. Compared to IP administration, the oral bioavailabilty of FTS was 69.5% for oral CMC and 55% for oral corn oil suspensions, while clearance and volume of distribution were higher in both oral preparations. The orally administered salirasib inhibited panc-1 tumor growth in a dose dependent manner (67% reduction in tumor weight at the highest dose, P < 0.002 vs. control, n = 10 mice per group) and at a 40 mg/kg daily dose was synergistic with gemcitabine (83% increase in survival rate, n = 8 mice per group). Salirasib exhibits good bioavailabilty after oral administration, as determined by a highly sensitive method for quantification in plasma. The orally available Ras inhibitor salirasib inhibited growth in nude mice, and may thus be considered for clinical trials.


Reversible carotid artery narrowing in morning glory disc anomaly.
A 14-year-old boy with morning glory disc anomaly (MGDA) and normal visual and neurologic function displayed marked carotid artery narrowing on magnetic resonance angiography (MRA). This narrowing disappeared on a follow-up MRA six months later. Optic coherence tomography and scanning laser polarimetry disclosed a normal retinal nerve fiber layer in the eye with MGDA. MGDA has been reported in association with irreversible carotid artery stenosis leading to moya moya disease. This case suggests that mild cases of MGDA may be associated with reversible carotid artery narrowing owing to vasospasm.


Severe Acute Respiratory Distress Syndrome (ARDS) or Severely Increased Chest Wall Elastance?
Esophageal manometry can be used to calculate transpulmonary pressures and optimize ventilator settings accordingly. We present the case of a 31-year-old male patient with ataxia-telangiectasia (Louis-Bar syndrome) and a BMI of 20 kg/m2, admitted to our intensive care unit for coronavirus disease 2019 (COVID-19) pneumonia. The patient soon required mechanical ventilation; however, there was very poor respiratory system compliance. Cholecystitis complicated the clinical course, and veno-venous extracorporeal membrane oxygenation (ECMO) was initiated as gas exchange deteriorated. Esophageal manometry was introduced and revealed severely increased intrathoracic pressure and chest wall elastance.


Role of Microglia and Astrocytes in Spinal Cord Injury Induced Neuropathic Pain.
Spinal cord injuries incite varying degrees of symptoms in patients, ranging from weakness and incoordination to paralysis. Common amongst spinal cord injury (SCI) patients, neuropathic pain (NP) is a debilitating medical condition. Unfortunately, there remain many clinical impediments in treating NP because there is a lack of understanding regarding the mechanisms behind SCI-induced NP (SCINP). Given that more than 450,000 people in the United States alone suffer from SCI, it is unsatisfactory that current treatments yield poor results in alleviating and treating NP. In this review, we briefly discussed the models of SCINP along with the mechanisms of NP progression. Further, current treatment modalities are herein explored for SCINP involving pharmacological interventions targeting glia cells and astrocytes. The studies presented in this review provide insight for new directions regarding SCINP alleviation. Given the severity and incapacitating effects of SCINP, it is imperative to study the pathways involved and find new therapeutic targets in coordination with stem cell research, and to develop a new gold-standard in SCINP treatment.


Student nurse stress in the preceptorship experience.
Student nurses appear to experience significantly more stress during their academic preparation than they do during the first year of employment. Preceptorship is among the most stressful of student experiences. It is within the context of a challenging and at times daunting work environment that two complete strangers (preceptor and student) strive to accommodate one another within a professional capacity. If the relationship between preceptor and student is less than successful, not only can it be frustrating and disheartening, but it can result in student stress and disillusionment about nursing and an inability to integrate and learn. Using a hypothetical case, the authors discuss the importance of student assessment, close communication between faculty and preceptors, and quick responses to student stress as a means by which to circumvent the serious potential of student burnout in the practice setting.


Decisions at the end of life: Catholic tradition.
Medical decisions regarding end-of-life care have undergone significant changes in recent decades, driven by changes in both medicine and society. Catholic tradition in medical ethics offers clear guidance in many issues, and a moral framework accessible to those who do not share the same faith as well as to members of its faith community. In some areas, a Catholic perspective can be seen clearly and confidently, such as in teachings on the permissibility of suicide and euthanasia. In others, such as withdrawal of nutrition and hydration, the Church does not yet speak with one voice and has not closed out the discussion. Yet, it is not in the teaching on individual issues that a Catholic moral tradition offers the most help and comfort, but in its account of what it means to lead a life in Christ, and to prepare for a Christian death. As in the problem of pain and suffering, it is the spiritual support more than the ethical guidance that helps both patients and physicians bear the unbearable and fathom the unfathomable.


Protective effects of lovastatin on vascular endothelium injured by low density lipoprotein.
To examine protective effects of lovastatin, an inhibitor of 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase, on endothelial dysfunction induced by a single intravenous injection of natural low density lipoprotein (n-LDL) and analyze the possible action mechanism of lovastatin. Rats were treated by intraperitoneal injection with lovastatin at dose of 2 or 4 mg/kg body weight once daily for 7 d, and on d 6 a single injection of n-LDL 4 mg/kg was given by sublingual vein. Forty eight hours after injection of n-LDL, the descending thoracic aorta of rats was taken. Acetylcholine (ACh)-induced endothelium-dependent relaxation (EDR) and sodium nitroprusside (SNP)-induced endothelium-independent relaxation of aortic rings were examined in vitro. Malondialdehyde (MDA), nitric oxide (NO), and activity of superoxide dismutase (SOD) as well as level of lipid in serum were analyzed. A single injection of n-LDL inhibited ACh-induced EDR compared with normal control group (maximal relaxation rate: 69.5 %+/-1.2 % vs 91.0 %+/-1.2 %, P<0.05), decreased serum NO level [(7.0+/-0.5) micromol/L vs (11.2+/-0.9) micromol/L, P<0.05] and serum SOD activity [(371+/-16) kNU/L vs (405+/-18) kNU/L, P<0.05] and elevated serum MDA level [(5.4+/-0.5) micromol/L vs (3.0+/-0.8) micromol/L, P<0.05]. Compared with n-LDL treated group, lovastatin 2 and 4 mg/kg increased EDR( maximal relaxation rate 82.9 %+/-0.5% and 83.7 %+/-0.7 % vs 69.5 %+/-1.2 %, P<0.05) and elevated NO level [(11.0+/-0.7) and (11.2+/-0.8) micromol/L vs (7.0+/-0.5) micromol/L, P<0.05], increased SOD activity [(402+/-15) and (408+/-25) kNU/L vs (371+/-16) kNU/L, P<0.05], and reduced serum MDA level [(3.3+/-0.6) and (3.5+/-0.4) micromol/L vs (5.4+/-0.5) micromol/L, P<0.05]. But sodium nitroprusside-induced endothelium-independent relaxation and the level of serum lipid in both saline+LDL group and lovastatin-treated group had no marked alteration. Lovastatin was able to protect vascular endothelium from dysfunction induced by a single injection of n-LDL.


Analysis of bacterial community structure in the natural circulation system wastewater bioreactor by using a 16S rRNA gene clone library.
A variety of physical and chemical parameters are routinely monitored during operation of the Natural Circulation System, a wastewater purification bioreactor in which only natural materials and no synthetic chemicals are used. However, the microbial community structures existing in the Natural Circulation System have not been well characterized. Thus, bacterial community structure and composition in this system were studied using clone library analysis of 16S ribosomal RNA genes amplified using PCR with universal bacterial primer sets. The PCR products were then subcloned into the pGEM-T vector. Each unique restriction fragment length polymorphism pattern, created by using two pairs of restriction endonucleases, was designated as an operational taxonomic unit (OTU). The Natural Circulation System comprises five tanks, the second and third of which play a major role in the bioreactor. Clone library pro-files and principal component analysis revealed differences in the bacterial community structures of the second (anaerobic chamber) and the third (aerobic chamber) tanks. However, the beta-proteobacteria, Bacteroidetes/ Chlorobi and gamma-proteobacteria groups were dominant in both tanks. Bacterial composition was more complex in the second tank (107 OTUs) than in the third tank (68 OTUs). Of a total of 154 OTUs in the clone libraries, only 21 were common to the two tanks. The results obtained in this study should provide important information for future research into and management of the Natural Circulation System wastewater bioreactor.


The effects of fasting and refeeding healthy volunteers on serum folate levels.
Fasting serum folate levels are commonly used in assessing folate status and also in estimating the bioavailability of synthetic folic acid and food folate. Previous work has shown that serum folate more than doubles in concentration during a 48 hour fast. Following a 24-hour standardisation procedure, serum and urinary folate levels were measured in nine healthy female volunteers fasting for 36 hours and in the first 6 hours during refeeding. Serum folate concentration increased from a mean of 14.8 ng/ml to 29.3 ng/ml during the 36 hour fast and fell to 22.1 ng/ml during the 6 hour refeeding period. The rise in serum folate concentration during the fast was negatively related to serum folate concentration at 0 hours fasting. It is hypothesised that the enterohepatic recirculation of folate plays an important role in the underlying physiological mechanism. These findings highlight the need to standardise energy intake to control for hepatic influences on folate metabolism in future studies assessing the bioavailability of folate in food.


Continuous subcutaneous insulin infusion.
Use of CSII requires care by skilled professionals, careful selection of patients, meticulous patient monitoring, and thorough patient education. Insulin pumps prescribed by a physician within these guidelines are a part of treatment and should be covered by the usual payment mechanisms.


Herbaspirillum psychrotolerans sp. nov., a member of the family Oxalobacteraceae from a glacier forefield.
A novel psychrotolerant, Gram-negative, shiny white, curved-rod-shaped, facultatively anaerobic bacterium PB1(T) was isolated from a soil sample collected from a glacier forefield of the Larsemann Hills, East Antarctica. Isolate PB1(T) has catalase and low urease activity and hydrolyses gelatin and starch. Strain PB1(T) is able to grow between -5 °C and 30 °C with optimum growth at 14-20 °C. Glycerol, dl-arabinose, d-xylose, d-galactose, d-fructose, d-lyxose, d-fucose and potassium gluconate are used as sole carbon sources. The major quinone is ubiquinone Q-8. The major fatty acids (>10%) for PB1(T) are C(16:0) (19.1%), C(16:1)ω7cis (44.6%) and C(18:1)ω7cis (16.2%). The major polyamines are putrescine [54.9 µmol (g dry weight)(-1)] and 2-hydroxy putrescine [18.5 µmol (g dry weight)(-1)]. DNA G+C content is 62.5 mol%. Strain PB1(T) is phylogenetically related to species of the genus Herbaspirillum, with highest 16S rRNA gene sequence similarities to Herbaspirillum canariense (97.3%), Herbaspirillum aurantiacum (97.2%), Herbaspirillum soli (97.2%) and Herbaspirillum frisingense (97.0%). The DNA-DNA relatedness values were below 30% between PB1(T) and the type strains of Herbaspirillum canariense, Herbaspirillum aurantiacum and Herbaspirillum soli. The different geographical origin of strain PB1(T) from its closest phylogenetic relatives resulted in different phenotypic and genotypic specifications, whereby strain PB(T) represents a novel species of the genus Herbaspirillum, for which the name Herbaspirillum psychrotolerans is proposed. The type strain is PB1(T) (DSM 26001(T) =LMG 27282(T)).


The prevalence of obesity and elevated liver enzymes in children at a university gastroenterology clinic.
Obesity has been recognized as a risk factor for liver disease. We evaluated the prevalence of obesity and elevated liver transaminases in children referred to the Pediatric Gastroenterology Clinic at the Marshall University Joan C. Edwards School of Medicine in Huntington. In a retrospective chart analysis the following data were collected: demographic, anthropometric, Body Mass Index, liver enzymes, medications, and final diagnosis. Of the 2550 charts reviewed, 540 children (21%) were obese, 399 (16%) were overweight, and 1,611 (63%) had normal weight. Liver enzymes were recorded in 902 children (35%). Elevated enzymes were found in 79 of the children (8.7%), but only 31 were appropriate for final calculation. Elevated liver enzymes were significantly higher in obese children compared to overweight or normal weight children. Over 30% of our children were obese or overweight. Obesity is a risk factor for elevated liver enzymes and early assessment is recommended.


Ontogenesis of the omasum: a comparative analysis of the Merino sheep and Iberian red deer.
The aim of this study is to describe differences in the ontogenesis of the omasum in sheep (domestic ruminant) and deer (wild ruminant). A total of 50 embryos and fetuses of Merino sheep and 50 Iberian deer were used, from the first stages of prenatal life until birth. For the study, the animals were divided into five experimental groups according to the most relevant histological characteristics. The appearance of the omasum from the primitive gastric tube was earlier in sheep (22% gestation, 33 days) than in deer (25% gestation, 66 days). In both cases it displayed a primitive epithelium of a stratified, cylindrical, non-ciliary type. The appearance of four laminae of different sizes was always earlier in sheep than deer. At around 36% gestation in sheep (53 days) and 36% (97 days) in deer, the omasum consisted of 4 clearly-differentiated layers: mucosa (with epithelial layer and lamina propria), submucosa, tunica muscularis and serosa. The temporal order of appearance of the four order laminae and omasal papillae was always earlier in sheep than deer. The tegumentary mucosa of the omasum was without secretion capability in the first embryonic phases. From 67 days (26% gestation) the neutral mucopolysaccharides appeared in deer and at 46 days (30% gestation) in sheep. In both cases they continued to decrease until birth, this decrease being more pronounced in deer. Finally, the presence of neuroendocrine and glial cells was detected in deer at earlier stages than in sheep.


Relación entre la medición por tomografía y ecografía del diámetro de la vaina del nervio óptico como estimador no invasivo de la presión intracraneal.
To compare the ONSD measured by ultrasound and tomography in patients with a diagnosis of intracranial hypertension. Prospective, transversal, observational, analytical study. 105 patients were included, divided into two groups: healthy (control group) and patients presenting clinical data of intracranial hypertension (study group). ONSD was measured by ultrasound and tomography. The Kruskal-Wallis test was used to compare the ONSD between the patients, and the Spearman test was used to assess the correlation between USG and CT. A value of p <0.05 was considered statistically significant. Of the 105 patients, 58.1% were men and 41.9% women. The study group included 14 patients with TBI, CVD, intracranial neoplasia, or neuroinfection. The highest median of ONSD by Ultrasound was in the CVD group, followed by TBI, neoplasia and neuroinfection and the lowest was in the control group (7.5, 7.0, 6.8, 6.8 and 5.2 mm respectively); these differences being statistically significant (p < 0.001). In the correlation analysis between Ultrasound and CT, a good statistically significant positive correlation was found (rho = 0.893, p < 0.001). The ultrasound evaluation of ONSD has proven to be a reliable test for the diagnosis and non-invasive monitoring of intracranial hypertension.


Template specific inhibitor of mammalian DNA polymerases.
A study of the inhibition of mouse cellular DNA polymerases by poly-nucleotides and their vinyl analogs is presented. Poly(dT)-directed poly(dA) synthesis by representatives of all three classes of cellular DNA polymerase could be completely inhibited by poly(9-vinyladenine), although higher concentrations were required in the case of the gamma class enzyme. Studies on the mechanism of the inhibition using the alpha class DNA polymerase and different templates showed that the enzyme activity was inhibited in all cases where base-pairing between the vinyl polymer and the template occurred; poly(9-vinyladenine) did not interfere with the replication of templates to which it does not bind. The inhibition occurred shortly after addition of poly(9-vinyladenine) to ongoing reactions, yet the enzyme was not displaced from the template - primer complex.


Cavβ3 Regulates Ca2+ Signaling and Insulin Expression in Pancreatic β-Cells in a Cell-Autonomous Manner.
Voltage-gated Ca2+ (Cav) channels consist of a pore-forming Cavα1 subunit and auxiliary Cavα2-δ and Cavβ subunits. In fibroblasts, Cavβ3, independent of its role as a Cav subunit, reduces the sensitivity to low concentrations of inositol-1,4,5-trisphosphate (IP3). Similarly, Cavβ3 could affect cytosolic calcium concentration ([Ca2 +]) in pancreatic β-cells. In this study, we deleted the Cavβ3-encoding gene Cacnb3 in insulin-secreting rat β-(Ins-1) cells using CRISPR/Cas9. These cells were used as controls to investigate the role of Cavβ3 on Ca2+ signaling, glucose-induced insulin secretion (GIIS), Cav channel activity, and gene expression in wild-type cells in which Cavβ3 and the IP3 receptor were coimmunoprecipitated. Transcript and protein profiling revealed significantly increased levels of insulin transcription factor Mafa, CaMKIV, proprotein convertase subtilisin/kexin type-1, and nitric oxide synthase-1 in Cavβ3-knockout cells. In the absence of Cavβ3, Cav currents were not altered. In contrast, CREB activity, the amount of MAFA protein and GIIS, the extent of IP3-dependent Ca2+ release and the frequency of Ca2+ oscillations were increased. These processes were decreased by the Cavβ3 protein in a concentration-dependent manner. Our study shows that Cavβ3 interacts with the IP3 receptor in isolated β-cells, controls IP3-dependent Ca2+-signaling independently of Cav channel functions, and thereby regulates insulin expression and its glucose-dependent release in a cell-autonomous manner.


Interpreting the language of breathlessness.
Janelle Yorke and Anne-Marie Russell discuss the sensation of breathlessness and the language used by patients and healthcare professionals to describe it.


Association of cannabis use disorder with cardiovascular diseases: A two-sample Mendelian randomization study.
The use of cannabis has increased globally due to more regions decriminalizing marijuana use for therapeutic and recreational aims. Several observational studies have revealed that cannabis use is associated with an increased risk of adverse cardiovascular pathologies and diseases. Nevertheless, the causal associations between cannabis use and cardiovascular diseases remain unclear. Hence, we performed single-variable and multivariable Mendelian randomization (MR) to evaluate the association between cannabis use disorder and various cardiovascular diseases. Summary statistics were collected from the largest-to-date genome-wide association studies (GWAS) of cannabis use disorder. The 12 SNPs for cannabis use disorder were used as instrumental variables in this study. MR estimates were pooled using a random-effects inverse-variance weighted (IVW) method. Simple median and weighted median methods were conducted as sensitivity analyses. The genetic liability to cannabis use disorder was associated with an augmented risk of coronary artery disease, myocardial infarction, atrial fibrillation, heart failure, deep venous thrombosis, pulmonary embolism, and stroke. Except for stroke, the results were inconsistent in the sensitivity analyses. The overall patterns for the associations of cannabis use disorder with atrial fibrillation, heart failure, pulmonary embolism and stroke remained in multivariable MR analyses adjusting for potential mediators, including smoking, alcohol, body mass index, blood lipid, type 2 diabetes, hypertension, and depression. However, the association with coronary artery disease, myocardial infarction, and deep venous thrombosis did not persist in multivariable MR analyses. Mediation analysis demonstrated that smoking, body mass index, low-density lipoprotein, hypertension, and depression have more significant mediation effects, which suggests that these factors partly mediate the link from cannabis use disorder to coronary artery disease, myocardial infarction, and deep venous thrombosis. The genetic liability to cannabis use disorder was associated with a higher risk of atrial fibrillation, heart failure, pulmonary embolism, and stroke. The evidence for the association between cannabis use disorder, coronary artery disease, myocardial infarction, and deep venous thrombosis was weak. Hence, future use of cannabis for therapeutic and recreational aims should consider its potential impact on cardiovascular diseases.


An algorithm-directed two-component library synthesized via solid-phase methodology yielding potent and orally bioavailable p38 MAP kinase inhibitors.
Previously we reported the identification of RPR200765A, a potent orally bioavailable pyridine-imidazole inhibitor of p38 mitogen-activated protein (MAP) kinase which suppressed paw swelling and joint pathology in streptococcal cell wall-induced arthritis. Herein, we report the use of solid-phase combinatorial organic synthesis for the parallel processing of a related pyrimidine-imidazole-based library with two points of structural variability. We report also that the application of a computer algorithm, the Monte Carlo Monomer Selection, maximized both the combinatorial synthetic efficiency and the bioavailability of the final compounds. In conjunction with the synthetic protocols, the polymer-supported quench technique was applied to the purification of the final compounds. Through rapid evaluation of the library using a p38 kinase assay and permeability assays, it was possible to identify a number of potent and orally bioavailable p38 MAP kinase inhibitors suitable for further biological investigation.


Functions of endothelin-1 in apoptosis and migration in hepatocellular carcinoma.
Hepatocellular carcinoma (HCC) is the second leading cause of cancer mortality in China and the third leading cause of cancer mortality worldwide. The mechanisms involved in the development and progression of HCC are not well understood. In the present study, the functions of endothelin-1 (ET-1) in HCC were studied and its underlying mechanisms were investigated. ET-1, B-cell lymphoma 2 (Bcl-2), Bcl-2 related protein 4 (Bax), matrix metalloproteinase (MMP)-2 and MMP-9 expression was measured by reverse transcription-quantitative polymerase chain reaction and western blotting. Cell proliferation was measured via Cell Counting kit-8 assay. Flow cytometry was performed for cell cycle and apoptosis analysis. Migration was measured via Transwell assay. The results demonstrated that ET-1 expression significantly increased in HCC tissues compared with the normal tissues of patients in The Cancer Genome Atlas dataset (P<0.01). Furthermore, downregulation of ET-1 was able to significantly inhibit cell proliferation and growth in vitro (P<0.01) and in vivo (P<0.01), and induce cell cycle arrest (P<0.05) and apoptosis (P<0.01) in the HCC SMMC-7721 cell line. Bioinformatics analysis demonstrated that the cell apoptosis signaling pathway was activated by ET-1. The ratio of B-cell lymphoma (Bcl-2) -related protein 4 (Bax)/Bcl-2 was significantly increased by downregulation of ET-1 (P<0.01). ET-1 downregulation also inhibited migration of SMMC-7721 cells (P<0.05) via decreasing levels of matrix metalloproteinase (MMP) -2 (P<0.05) and MMP-9 (P>0.05). These results suggest that ET-1 may be able to affect the apoptosis and migration of HCC cells via modulation of the Bax/Bcl-2 ratio and expression levels of MMP-2 and MMP-9, which indicates that ET-1 maybe a potential novel target for HCC treatment.


Lens thickness and angle-closure glaucoma. A comparative oculometric study in South African Negroes and Danes.
A group of Negro patients without ocular disease ("normal" group), a group of Negroes with angle closure glaucoma and a normal Danish population are compared in terms of anterior chamber depth, less thickness, vitreous body length and axial length of the globe. It is noted that the normal black population has a significantly thinner lens than the black population with angle closure glaucoma and the normal Danish population. The latter groups have a lens thickness which is similar. This finding is discussed in relation to the lower incidence of acute closure attacks in the black population. Attacks of angle closure in this populatin are generally of the chronic type.


Performance Assessment of Epidermal Growth Factor Receptor Gene Sequencing According to Sample Size in Daily Practice Conditions.
Lung carcinoma is the main cause of cancer death worldwide. Adenocarcinoma molecular biomarkers have been discovered, and targeted therapies have been developed with encouraging results. The epidermal growth factor receptor gene is one of these biomarkers. Exons 18 to 21 should be studied in patients with advanced adenocarcinoma, who are candidates for treatment with tyrosine kinase inhibitors. The objective was to compare the performance of the determination in large and small samples in daily practice conditions, trying to adjust to published consensus guidelines. A retrospective observational study of 141 cases was carried out, with exons 19 and 21 sequencing. Sample size (small vs. large), including number of satisfactory polymerase chain reaction (PCR), sequencing, deletions, and mutations, were evaluated. In small biopsies, sample type, fragment number, and percentage of tumor per sample were analyzed. The results shown 114/141 (80.8) cases that met selection criteria; 60/114 (53%) were large (surgical) and 54/114 (47%) were small samples (19/54 endoscopic, 17/54 fine needle aspiration clots, 4/54 lymph nodes, 14/54 core and other). All large samples were satisfactory PCR, 56/60 (93%) satisfactory sequencing, and 12/56 (21%) had deletions in exon 19. Small samples were satisfactory PCRs in 50/54 (93%) cases, and satisfactory sequencing in 35/50 (65%), 8/35 (23%) showed alterations in exon 19, and 1/35 (3%) in exon 21. In conclusion, the proportion of samples unfit for the study of the epidermal growth factor receptor gene mutational status increased from 7% in large samples to 35% in small ones. Nineteen small samples were inconclusive, with cell blocks predominating, 10/19 (53%).


Repeated mechanical loading enhances the expression of Indian hedgehog in condylar cartilage.
Indian hedgehog (Ihh) acts as a mechanotransduction mediator that converts mechanical strain into cellular proliferation and cartilage formation in mandibular condylar cartilage. The aim of this study was to examine the effect of repeated mechanical strain on the level of expression of Ihh and type II collagen mRNA in condylar growth. Two hundred and eighty 35 days old Sprague-Dawley rats were divided into 10 experimental and 10 control groups. Repeated mechanical loading was applied by advancing the mandible in a stepwise manner by fitting a stepwise bite jumping appliance. Animals were sacrificed together with their matched controls on 10 different time points. Total RNA was extracted from condylar cartilage immediately after dissection. Ihh and type II collagen mRNA was quantified using real-time PCR. In the experimental group, Ihh mRNA increased significantly on experimental day 7. Upon the second advancement, another peak was elicited 7 days later. Type II collagen showed a significant increase on days 21 and 44 of advancement. This indicated that mechanical loading in a repeated manner, triggers the expression of Ihh which in turn increases the number of replicating mesenchymal cells as well as the amount of the cartilage formed. Taken together these events increase condylar growth.


Currents in defective coupled ratchets.
Transport phenomena in a one-dimensional system of interacting particles is studied. This system is embedded in a periodic and left-right asymmetric potential driven by a force periodic in time and space. When the density (number of particles per site) is an integer, directional current of the particles is collective; that is, it involves the whole system since all the sites are equivalents. On the other hand, when the system has a defect, a new localized or noncollective current appears due to the migration of defects from one site to another. We show here how this "defective" (defects generated) current can be controlled by white noise.


[Evaluation of phagocytosis, bactericidal effect, and cytotoxicity of Pasteurella haemolytica and Pasteurella multocida in bovine alveolar macrophages].
Evaluation of phagocytosis and bactericidal effect of Pasteurella haemolytica and P. multocida was conducted on bovine alveolar macrophages freshly obtained through bronchioalveolar washings from live animals. Cytotoxic activity of these bacteria on the alveolar macrophages was evaluated through the simple visual assay in microplates, using bovine blood leukocytes as a comparative target cell. In order to evaluate phagocytosis the following variables were considered P. haemolytica and P. multocida (independently) in contact with alveolar macrophages, P. haemolytica and P. multocida in suspension as a positive control of bacterial growth, and RPMI-1640 medium alone, as a negative control of bacterial growth. To measure bactericidal capacity, bacteria were incubated with plastic adhered alveolar macrophages at 30 minutes and 3 hours intervals. Samples incubated 30 minutes were taken as phagocytosis-base readings and at the 3 h interval to evaluate bactericidal capacity of the alveolar macrophages on phagocytized bacteria. Reading of the samples of each evaluation was conducted in a spectrophotometer a 380 nm. Phagocytosis results indicated that bacterial proliferation was higher when bacteria were alone as compared when they were with alveolar macrophages (p < 0.05). Bactericidal capacity of the macrophages was efficient because bacterial numbers were higher in the first evaluation as compared to the second (p < 0.05). It was demonstrated that the cytotoxic effect of P. haemolytica was more severe on blood leukocytes as compared to alveolar macrophages (p < 0.05). There was no evidence of P. multocida cytotoxicity on the evaluated cells. With the development of this technique for the obtention of alveolar macrophages and using spectrophotometry for the phagocytosis and bactericidal effect evaluations, numerous variables and samples can be tested, such as opsonized bacteria or to measure the behaviour of alveolar macrophages infected with different agents involved in the bovine pneumonic complex.


Intravitreal and plasma cidofovir concentrations after intravitreal and intravenous administration in AIDS patients with cytomegalovirus retinitis.
This study was undertaken to evaluate the intravitreal and plasma concentrations of cidofovir (HPMPC) after intravitreal and intravenous administration in AIDS patients with cytomegalovirus retinitis. Cohort series; undiluted vitreous and blood were collected from 9 patients at the time of pars plana vitrectomy. Vitreous samples from 9 eyes of 9 patients and plasma samples from 4 patients were assayed with high-performance liquid chromatography to determine cidofovir levels. The only eye that had a detectable vitreous concentration (673.7 ng/ml) was injected with 20 microg 24 hours prior to the surgery. The remaining samples including plasma were below the detection point of the assay (100 ng/ml) and were injected between 5 and 40 days prior to sampling. The intravitreal concentration of cidofovir in humans is consistent with pharmacokinetics data in laboratory animals, and suggests that the long duration of antiviral effect (1-3 months) in clinical trials is due to a prolonged intracellular half-life in retinal tissue.


A DNA-Based Assessment of the Phylogenetic Position of a Morphologically Distinct, Anchialine-Lake-Restricted Seahorse.
Isolated populations provide special opportunities to study local adaptation and incipient speciation. In some cases, however, morphological evolution can obscure the taxonomic status of recently founded populations. Here, we use molecular markers to show that an anchialine-lake-restricted population of seahorses, originally identified as Hippocampus reidi, appears on the basis of DNA data to be Hippocampus erectus We collected seahorses from Sweetings Pond, on Eleuthera Island, Bahamas, during the summer of 2014. We measured morphological traits and sequenced 2 genes, cytochrome b and ribosomal protein S7, from 19 seahorses in our sample. On the basis of morphology, Sweetings Pond seahorses could not be assigned definitively to either of the 2 species of seahorse, H. reidi and H. erectus, that occur in marine waters surrounding the Bahamas. However, our DNA-based phylogenetic analysis showed that the Sweetings Pond fish were firmly nested within the H. erectus clade with a Bayesian posterior probability greater than 0.99. Thus, Sweetings Pond seahorses most recently shared a common ancestor with H. erectus populations from the Western Atlantic. Interestingly, the seahorses from Sweetings Pond differ morphologically from other marine populations of H. erectus in having a more even torso to tail length ratio. The substantial habitat differences between Sweetings Pond and the surrounding coastal habitat make Sweetings Pond seahorses particularly interesting from the perspectives of conservation, local adaptation, and incipient speciation.


High and low rearing subgroups of rats selected in the open field differ in the activity of K+-stimulated p-nitrophenylphosphatase in the hippocampus.
Na(+)/K(+)-adenosinetriphosphatase (Na(+)/K(+)-ATPase) is of paramount importance for the proper functioning of the organism. The enzyme is involved in several aspects of brain function, such as the repolarization of the neuronal membranes and neurotransmitters uptake/release. Therefore, individual differences in the activity of brain Na(+)/K(+)-ATPase may result in differences in the functioning of the brain, which, in consequence, could lead to behavioral divergences. Individual differences in rearing, an exploratory behavior, have been shown to be genetically determined. In rats, the inhibition of the activity of Na(+)/K(+)-ATPase was reported to induce changes in the exploratory behavior. The aim of this work was to verify if subgroups of rats selected according to the number of rearings (high and low rearing subgroups) in the open field test differ in the activity of Na(+)/K(+)-ATPase in brain regions. Adult, male outbred Wistar rats were selected in the open field test according to the number of rearings in subgroups of high (HR) and low (LR) rearing responders. After a rest of about 20 days after the open field session, HR and LR rats were sacrificed. In the first experiment, frontal cortex, striatum, brainstem, hippocampus and the amygdala (including the overlying limbic cortex) were dissected. The reaction of dephosphorylation of Na(+)/K(+)-ATPase (K(+) stimulated p-nitrophenylphosphatase) was assayed in homogenates rich in synaptosomes. The results obtained showed a statistically significant higher activity of K(+)p-nitrophenylphosphatase only in the hippocampus of HR subgroup of rats. This result was replicated in two other subsequent experiments with different HR and LR subgroups of rats selected at different times of the year. Our data suggest that the difference in the activity of Na(+)/K(+)-ATPase in the hippocampus is innate and is involved in the expression of the rearing behavior.