Revealing Different Lung Metastatic Potentials Induced Metabolic Alterations of Hepatocellular Carcinoma Cells via Proton Nuclear Magnetic Resonance Spectroscopy.
Background: Hepatocellular carcinoma (HCC) causes death mainly by disseminated metastasis progression and major challenge of clinical management is to distinguish lethal metastatic stage from indolent stage. It is shown that metastatic progression is closely related to cellular metabolism. But detailed metabolic alterations and molecular mechanisms still kept unclear between subtypes of different lung metastatic potentials. Methods: The current work used NMR-based metabolomics in the study of HCC cells with high malignancy but differed in lung metastatic potentials. Cell extracts and cultured media from MHCC97L and MHCC97H were utilized to reveal metabolic alterations related to metastatic potentials. Multivariate analyses were performed to identify characteristic metabolites which were used subsequently to draw the map of relative biochemical pathways by combining KEGG database. Results: The NMR spectra of both MHCC97L and MHCC97H include various signals from necessary nutritional components and metabolic intermediates. A series of characteristic metabolites were determined from both cell extracts and media. The ability on nutrient uptake varied from cell lines. Most of amino acids decreased in high metastatic cell line, so altered amino acid metabolisms and energy metabolism were revealed in high metastatic MHCC97H cell line. The majority pathways involved six essential amino acids in which the observed branched-chain amino acids together with lysine contributed to biosynthesis or degradation. Basically MHCC97H cell line could induce more active events than that of MHCC97L to progress to high metastasis with certain molecular events. Characteristic metabolites-derived classifiers performed robustly during prediction and confirmed their critical role in supporting metastasis progression. Conclusions: Our results provide evidence that NMR-metabolomics analyses of cells are able to understand metastatic characteristics accountable for biological properties. The proposed characteristic metabolites will help to understand HCC metastatic characterizations and may be filtered as potential biomarkers.


Effects of Social Media on Consumer Information Seeking and Purchase Intention of Oral Health Care Products Among Adults.
Purpose Social media has created an environment providing readily available information that impacts how individuals seek information and make product decisions. The purpose of this study was to examine adult consumer use of social media for information seeking and the effect on intention to purchase oral health care products.Methods A cross-sectional study design with a non-probability sample of adult consumers recruited through crowdsourcing with Amazon's Mechanical Turk (MTurk) was used to examine the effect of social media on purchasing of oral health products. Data collection included demographic, social media use, and the Information Acceptance Model (IACM) which measured information acceptance from friends and customer reviews from social media. Descriptive statistics, subgroup analysis of demographic variables, and multivariable linear regression were used to assess the relationship among variables.Results Participants (n=308) were mostly males (56.2%), average age 37 years, held bachelor's degrees (68.8%), from the southern United States, and had been to the dentist within the last two years (72.8%). Instagram (40.6%) was the most frequently used platform and social media was frequently used (36.7%) to obtain information on oral health care products. Findings from the IACM information subscale indicated high scores for information from friends and customer reviews (p<0.01). The information needs (p=0.049) and information adoption from friends (p=0.03) were slightly less for females when compared to males. Friends' recommendations (p=0.02) on social media had the greatest effect on intention to purchase among participants living in the Midwest. Multivariate regression models indicated information usefulness and information adoption had the greatest influence on purchase intention from friends (p<0.001), while information credibility, information usefulness, and information adoption had the greatest influence from customer reviews (p<0.001).Conclusion Results provided some initial insights into the way consumers may use social media for information about oral health products and the influences on them choosing to purchase the products. Oral health care providers can use this information to educate and guide patients to credible information sources in their search for oral health care products.


Medical Therapy Before Carotid Endarterectomy: Changes Over a 13-Year Period and Comparison Between Asymptomatic and Symptomatic Patients.
We evaluated the evolution of chronic medical therapy in patients admitted for carotid endarterectomy (CEA) over a 13-year period and to analyze the difference in medical treatment between symptomatic and asymptomatic patients. A retrospective study was conducted on patients treated between 2002 and 2015. The use of antiplatelets (acetylsalicylic acid [ASA], ticlopidine, and clopidogrel), oral anticoagulant therapy (OAT), statins and antihypertensives (angiotensin-converting enzyme inhibitors [ACE-I]/angiotensin receptor blockers [ARBs], β-blockers [BB]) administration was evaluated. During the study period, 852 CEAs were performed in 681 (79.9%) asymptomatic patients. Prescription rate increased significantly for ASA (+29.2%), clopidogrel (+10.3%), statins (+60.8%), ACE-I/ARBs (+31.1%), and BB (+19.3%; all Ptrend < .05). No significant modification was observed for ticlopidine and OAT (ticlopidine use was abandoned in the recent years, but this difference was not significant due to the small numbers). A lower medication intake was recorded for symptomatic patients when compared with asymptomatic patients, except for OAT and clopidogrel. Our analysis suggests that medical therapy has changed over the years for patients with carotid stenosis. Although this is a big step toward best medical therapy, preoperative drug therapy remains suboptimal in symptomatic patients.


Pre-certification: lessons from diagnostic imaging.
Recently, managed care has received a great deal of criticism from the public and press, some of it richly deserved. One of the areas of greatest concern is pre-certification requirements and possible denial of services. Experience with programs of this sort in diagnostic imaging suggests that, implemented correctly, the consultative interface created by a pre-certification program can be of tremendous value in physician education. Appropriate behavior modification programs need to be crafted with an educational rather than economic bias and the methodology not be totally abandoned simply because it has been misused by some. The pre-certification process can be used as a vehicle to disseminate information about rapidly emerging new therapies and technologies.


Pejorative testimony about opposing experts and colleagues: "fouling one's own nest".
Circumstances sometimes require expert witnesses under oath to express opinions about opposing experts or professional colleagues who work within their own professional, geographic, or organizational circle. This requirement poses some common problems that have not been well discussed in the literature. This article addresses some of those problems and suggests some useful solutions.


The green seaweed Ulva: a model system to study morphogenesis.
Green macroalgae, mostly represented by the Ulvophyceae, the main multicellular branch of the Chlorophyceae, constitute important primary producers of marine and brackish coastal ecosystems. Ulva or sea lettuce species are some of the most abundant representatives, being ubiquitous in coastal benthic communities around the world. Nonetheless the genus also remains largely understudied. This review highlights Ulva as an exciting novel model organism for studies of algal growth, development and morphogenesis as well as mutualistic interactions. The key reasons that Ulva is potentially such a good model system are: (i) patterns of Ulva development can drive ecologically important events, such as the increasing number of green tides observed worldwide as a result of eutrophication of coastal waters, (ii) Ulva growth is symbiotic, with proper development requiring close association with bacterial epiphytes, (iii) Ulva is extremely developmentally plastic, which can shed light on the transition from simple to complex multicellularity and (iv) Ulva will provide additional information about the evolution of the green lineage.


Cell lineage, cell death, and the developmental origin of identified serotonin- and dopamine-containing neurons in the leech.
The nervous system of the glossiphoniid leech includes segmentally iterated neurons that contain serotonin (5-HT) and dopamine. These have been investigated in Helobdella triserialis, Theromyzon rude, and Haementeria ghilianii. Five types of 5-HT neurons are identified by immunocytochemistry in the abdominal ganglia of the ventral nerve cord: the bilaterally paired Retzius, anteromedial, ventrolateral and dorsolateral neurons, and the unpaired posteromedial (pm) neuron. Three types of bilaterally paired dopamine neurons are identified by glyoxylic acid-induced fluorescence in the segmental body wall: MD, LD1, and LD2. Each left or right half of the segmental complement of the leech nervous system is known to develop from 6 distinct ectodermal primary blast cells (ns, nf, o, p, qs, and qf). To identify the blast cells of origin of the 5-HT and dopamine neurons, fluorescent cell lineage tracers were injected into the various precursors of the blast cells in early (stage 6) embryos. The embryos were then raised until their 5-HT and dopamine neurons could be scored (stage 11) for the presence or absence of lineage tracer. We find that the Retzius, anteromedial, and posteromedial 5-HT neurons are derived from the ns blast cell, while the ventrolateral and dorsolateral 5-HT neurons are derived from the nf blast cell. The unpaired pm 5-HT neuron arises as one of a bilateral pair of neurons, of which one later dies. Whether the left or right pm neuron survives in any given ganglion is the consequence of some form of competitive interaction between cells derived from the left and right n primary blast cells, possibly between the left and right pm neurons themselves. We find that, of the dopamine neurons, the LD1 neuron is derived from the o blast cell, the LD2 neurons from the p blast cell, and the MD neuron from one of the 2 kinds of q blast cells. These results show that the 5-HT and dopamine neurons arise from 5 different primary blast cells in a highly determinate manner, and they support the view that cells of a similar phenotype need not be closely related in the developmental cell lineage tree.


Androgen deprivation therapy, comorbidity, cancer stage and mortality from COVID-19 in men with prostate cancer.
Androgens facilitate entrance of the severe acute respiratory syndrome coronavirus 2 into respiratory epithelial cells, and male sex is associated with a higher risk of death from corona virus disease (COVID-19). Androgen deprivation therapy (ADT) could possibly improve COVID-19 outcomes. In a case-control study nested in the Prostate Cancer data Base Sweden (PCBaSe) RAPID 2019, we evaluated the association between ADT and COVID-19 as registered cause of death in men with prostate cancer. Each case was matched to 50 controls by region. We used conditional logistic regression to adjust for confounders and also evaluated potential impact of residual confounding. We identified 474 men who died from COVID-19 in March-December 2020. In crude analyses, ADT exposure was associated with an increased risk of COVID-19 death (odds ratio [OR] 5.05, 95% CI: 4.18-6.10); however, the OR was substantially attenuated after adjustment for age, comorbidity, prostate cancer characteristics at diagnosis, recent healthcare use, and indicators of advanced cancer (adjusted OR 1.25, 95% CI: 0.95-1.65). If adjustment has accounted for at least 85% of confounding, then the true effect could be no more than a 5% reduction of the odds for COVID-19 death. The increased mortality from COVID-19 in men with prostate cancer treated with ADT was mainly related to high age, comorbidity, and more advanced prostate cancer. There was no evidence to support the hypothesis that ADT is associated with improved COVID-19 outcomes.


ViSEAGO: a Bioconductor package for clustering biological functions using Gene Ontology and semantic similarity.
The main objective of ViSEAGO package is to carry out a data mining of biological functions and establish links between genes involved in the study. We developed ViSEAGO in R to facilitate functional Gene Ontology (GO) analysis of complex experimental design with multiple comparisons of interest. It allows to study large-scale datasets together and visualize GO profiles to capture biological knowledge. The acronym stands for three major concepts of the analysis: Visualization, Semantic similarity and Enrichment Analysis of Gene Ontology. It provides access to the last current GO annotations, which are retrieved from one of NCBI EntrezGene, Ensembl or Uniprot databases for several species. Using available R packages and novel developments, ViSEAGO extends classical functional GO analysis to focus on functional coherence by aggregating closely related biological themes while studying multiple datasets at once. It provides both a synthetic and detailed view using interactive functionalities respecting the GO graph structure and ensuring functional coherence supplied by semantic similarity. ViSEAGO has been successfully applied on several datasets from different species with a variety of biological questions. Results can be easily shared between bioinformaticians and biologists, enhancing reporting capabilities while maintaining reproducibility. ViSEAGO is publicly available on https://bioconductor.org/packages/ViSEAGO .


Bw4 ligand and direct T-cell receptor binding induced selection on HLA A and B alleles.
The HLA region is the hallmark of balancing selection, argued to be driven by the pressure to present a wide variety of viral epitopes. As such selection on the peptide-binding positions has been proposed to drive HLA population genetics. MHC molecules also directly binds to the T-Cell Receptor and killer cell immunoglobulin-like receptors (KIR). We here combine the HLA allele frequencies in over six-million Hematopoietic Stem Cells (HSC) donors with a novel machine-learning-based method to predict allele frequency. We show for the first time that allele frequency can be predicted from their sequences. This prediction yields a natural measure for selection. The strongest selection is affecting KIR binding regions, followed by the peptide-binding cleft. The selection from the direct interaction with the KIR and TCR is centered on positively charged residues (mainly Arginine), and some positions in the peptide-binding cleft are not associated with the allele frequency, especially Tyrosine residues. These results suggest that the balancing selection for peptide presentation is combined with a positive selection for KIR and TCR binding.


CD41-YFP mice allow in vivo labeling of megakaryocytic cells and reveal a subset of platelets hyperreactive to thrombin stimulation.
Development of a mouse line permitting live imaging of cells expressing CD41/GpIIb as a means to study megakaryopoiesis. The gene encoding yellow fluorescent protein (eyfp) was inserted by homologous recombination into embryonic stem cells at the start site of the gpIIb locus. A knockin mouse line, designated CD41-yellow fluorescent protein (YFP), was developed and was characterized by fluorescence microscopy and flow cytometry. Activity of YFP(+) platelets was determined by induction of P-selectin expression in response to thrombin stimulation. CD41-YFP mice contained YFP-labeled megakaryocytes and platelets, the proportions of which varied, depending on the genotype and individual animal, while lymphoid, myelomonocytic, and erythroid lineages were negative. In addition, a fraction of hematopoietic stem cells and intermediate progenitors expressed YFP at low levels. Crossing CD41-YFP mice with lysozyme green fluorescent protein and globin cyan fluorescent protein mice, followed by in vivo imaging of fetal liver, revealed megakaryocytic cells as a subset distinct from myeloid and erythroid cells. This experiment is also the first to show the distribution of three hematopoietic lineages in a minimally perturbed organ. Surprisingly, analysis of CD41-YFP platelets showed that the YFP(+) subset is more responsive to thrombin stimulation than the YFP(-) subset. Experiments aimed at determining the stability of the YFP(+) platelets showed that after lethal irradiation of CD41-YFP mice, the proportion of labeled platelets in the blood declines more rapidly than the bulk of the platelets. The newly developed mouse line should become useful not only for in vivo imaging experiments of megakaryocytes and platelets, but also for studies on platelet aging and function. Our irradiation experiments suggest that the YFP(+) platelets are enriched for newly made cells because YFP has a shorter half-life than platelets. Therefore, the finding that YFP(+) platelets are more responsive to thrombin stimulation raises the possibility that platelet activity decreases rapidly during physiological aging.


APOL1 risk variants in kidney transplantation: a modulation of immune cell function.
APOL1 G1 and G2 variants are established risk factors for nondiabetic kidney disease. The presence of two APOL1 risk variants in donor kidneys negatively impacts kidney allograft survival. Because of evolutionary pressure, the APOL1 risk variants have become common in people from Africa and in those with recent African ancestry. APOL1 risk variant proteins are expressed in kidney cells and can cause toxicity to these cells. In this issue of the JCI, Zhang, Sun, and colleagues show that recipient APOL1 risk variants negatively affect kidney allograft survival and T cell-mediated rejection rates, independent of donor APOL1 genotype or recipient ancestry. The authors provide evidence that APOL1 risk variants play an immunomodulatory role in T cells and NK cells in the setting of kidney transplantation. These findings have important clinical implications that require further investigation.


Communication: The electrostatic polarization is essential to differentiate the helical propensity in polyalanine mutants.
The folding processes of three polyalanine peptides with composition of Ac-(AAXAA)(2)-GY-NH(2) (where X is chosen to be Q, K, and D) are studied by molecular dynamics simulation in solvent of 40% trifluoroethanol using both polarized and unpolarized force fields. The simulations reveal the critical role of polarization effect for quantitative description of helix formation. When polarized force field is used, peptides with distinctive helical propensity are correctly differentiated and the calculated helical contents are in close agreement with experimental measurement, indicating that consideration of polarization effect can correctly predict the effect of sequence variation on helix formation.


Data-Driven Many-Body Models for Molecular Fluids: CO2/H2O Mixtures as a Case Study.
In this study, we extend the scope of the many-body TTM-nrg and MB-nrg potential energy functions (PEFs), originally introduced for halide ion-water and alkali-metal ion-water interactions, to the modeling of carbon dioxide (CO2) and water (H2O) mixtures as prototypical examples of molecular fluids. Both TTM-nrg and MB-nrg PEFs are derived entirely from electronic structure data obtained at the coupled cluster level of theory and are, by construction, compatible with MB-pol, a many-body PEF that has been shown to accurately reproduce the properties of water. Although both TTM-nrg and MB-nrg PEFs adopt the same functional forms for describing permanent electrostatics, polarization, and dispersion, they differ in the representation of short-range contributions, with the TTM-nrg PEFs relying on conventional Born-Mayer expressions and the MB-nrg PEFs employing multidimensional permutationally invariant polynomials. By providing a physically correct description of many-body effects at both short and long ranges, the MB-nrg PEFs are shown to quantitatively represent the global potential energy surfaces of the CO2-CO2 and CO2-H2O dimers and the energetics of small clusters, as well as to correctly reproduce various properties in both gas and liquid phases. Building upon previous studies of aqueous systems, our analysis provides further evidence for the accuracy and efficiency of the MB-nrg framework in representing molecular interactions in fluid mixtures at different temperature and pressure conditions.


Comparison of ex vivo and in vitro human fibroblast ageing models.
Several studies have analyzed modulation of gene expression during physiological ageing with interesting, but often contradictory results, depending on the model used. In the present report we compare age-related metabolic and synthetic parameters in human dermal fibroblasts (HDF) isolated from young and old subjects (ex vivo ageing model) and cultured from early up to late cumulative population doublings (CPD) (in vitro ageing model) in order to distinguish changes induced in vivo by the aged environment and maintained in vitro, from those associated with cell senescence and progressive CPD. Results demonstrate that fibroblasts from aged donors, already at early CPD, exhibit an impaired redox balance, highlighting the importance of this parameter during ageing, even in the presence of standard environmental conditions, which are considered optimal for cell growth. By contrast, several proteins, as those related to heat shock response, or involved in endoplasmic reticulum and membrane trafficking, appeared differentially expressed only during in vitro ageing, suggesting that, at high CPD, the whole cell machinery becomes permanently altered. Finally, given the importance of the elastic component for a long-lasting connective tissue structural and functional compliance, this study focuses also on elastin and fibulin-5 synthesis and deposition, demonstrating a close relationship between fibulin-5 and ageing.


Three-dimensional models of cancer for pharmacology and cancer cell biology: capturing tumor complexity in vitro/ex vivo.
Cancers are complex and heterogeneous pathological "organs" in a dynamic interplay with their host. Models of human cancer in vitro, used in cancer biology and drug discovery, are generally highly reductionist. These cancer models do not incorporate complexity or heterogeneity. This raises the question as to whether the cancer models' biochemical circuitry (not their genome) represents, with sufficient fidelity, a tumor in situ. Around 95% of new anticancer drugs eventually fail in clinical trial, despite robust indications of activity in existing in vitro pre-clinical models. Innovative models are required that better capture tumor biology. An important feature of all tissues, and tumors, is that cells grow in three dimensions. Advances in generating and characterizing simple and complex (with added stromal components) three-dimensional in vitro models (3D models) are reviewed in this article. The application of stirred bioreactors to permit both scale-up/scale-down of these cancer models and, importantly, methods to permit controlled changes in environment (pH, nutrients, and oxygen) are also described. The challenges of generating thin tumor slices, their utility, and potential advantages and disadvantages are discussed. These in vitro/ex vivo models represent a distinct move to capture the realities of tumor biology in situ, but significant characterization work still remains to be done in order to show that their biochemical circuitry accurately reflects that of a tumor.


Poorly differentiated carcinoma of unknown primary site: correlation of light microscopic findings with response to cisplatin-based combination chemotherapy.
We have previously reported complete responses and long-term survival in patients with metastatic poorly differentiated carcinoma (PDC) of unknown primary site who received intensive cisplatin-containing chemotherapy regimens. We reviewed the light microscopic specimens from 113 patients with PDC in an attempt to identify common histopathologic features in the chemotherapy-responsive subgroup, and to rule out the presence of previously unrecognized germ cell tumors. Relatively few diagnoses more specific than PDC could be made. We could identify no histopathologic features by light microscopy that distinguished responsive from unresponsive neoplasms. Only one patient was found to have a previously unrecognized yolk sac carcinoma, and in five other patients the possibility of a germ cell neoplasm was considered in the differential diagnosis by at least one reviewer. The remaining tumors had no histologic features suggestive of germ cell neoplasms. Ninety-six patients had received combination chemotherapy (89 with cisplatin-containing regimens); 27 patients (28%) achieved complete remission, and 16 remain free of disease at a median of 65 months after completion of therapy. Patients with PDC of unknown primary site who are responsive to cisplatin-containing chemotherapy regimens cannot be reliably identified by light microscopy. At present, all such patients should be considered for an empiric trial of chemotherapy with cisplatin-based regimens, since cure is achievable in a minority.


Prostate-specific amplification of expanded polyglutamine expression: a novel approach for cancer gene therapy.
For cancer gene therapy, it is of primary importance to develop a system to sufficiently and selectively express therapeutic genes in cancer cells. In this study, we showed that an approximately 5.3-kb promoter region of the prostate-specific antigen (PSA) gene can replicate the endogenous expression pattern, although its expression is very weak. We then developed a novel two-step transcriptional activation system in which the PSA promoter drives an artificial transcriptional activator, GAL4-VP16 fusion protein, and it in turn activates transgene expressions under the control of GAL4-responsive elements. By using this system, transgene expressions can be greatly augmented while maintaining prostate-specific expression. Finally, we applied this system to drive an expanded polyglutamine, a potent proapoptotic molecule, to induce apoptosis selectively in PSA-positive prostate cancer cells. This novel system would provide an ideal approach for cancer gene therapy applicable not only to prostate cancer but to other cancers as well.


Substitution mapping and characterization of brown planthopper resistance genes from indica rice variety, 'PTB33' (Oryza sativa L.).
Rice (Oryza sativa L.) yield is severely reduced by the brown planthopper (BPH), Nilaparvata lugens Stål, in Asian countries. Increasing resistance in rice against BPH can mitigate yield loss. Previous reports indicated the presence of three BPH resistance genes, BPH2, BPH17-ptb, and BPH32, in durable resistant indica rice cultivar 'PTB33'. However, several important questions remain unclear; the genetic locations of BPH resistance genes on rice chromosomes and how these genes confer resistance, especially with relationship to three major categories of resistance mechanisms; antibiosis, antixenosis or tolerance. In this study, locations of BPH2, BPH17-ptb, and BPH32 were delimited using chromosome segment substitution lines derived from crosses between 'Taichung 65' and near-isogenic lines for BPH2 (BPH2-NIL), BPH17-ptb (BPH17-ptb-NIL), and BPH32 (BPH32-NIL). BPH2 was delimited as approximately 247.5 kbp between RM28449 and ID-161-2 on chromosome 12. BPH17-ptb and BPH32 were located between RM1305 and RM6156 on chromosome 4 and RM508 and RM19341 on chromosome 6, respectively. The antibiosis, antixenosis, and tolerance were estimated by several tests using BPH2-NIL, BPH17-ptb-NIL, and BPH32-NIL. BPH2 and BPH17-ptb showed resistance to antibiosis and antixenosis, while BPH17-ptb and BPH32 showed tolerance. These results contribute to the development of durable BPH resistance lines using three resistance genes from 'PTB33'.


Vibrational states of a water molecule in a nano-cavity of beryl crystal lattice.
Low-energy excitations of a single water molecule are studied when confined within a nano-size cavity formed by the ionic crystal lattice. Optical spectra are measured of manganese doped beryl single crystal Mn:Be3Al2Si6O18, that contains water molecules individually isolated in 0.51 nm diameter voids within the crystal lattice. Two types of orientation are distinguished: water-I molecules have their dipole moments aligned perpendicular to the c axis and dipole moments of water-II molecules are parallel to the c-axis. The optical conductivity σ(ν) and permittivity ɛ'(ν) spectra are recorded in terahertz and infrared ranges, at frequencies from several wavenumbers up to ν = 7000 cm(-1), at temperatures 5-300 K and for two polarizations, when the electric vector E of the radiation is parallel and perpendicular to the c-axis. Comparative experiments on as-grown and on dehydrated samples allow to identify the spectra of σ(ν) and ɛ'(ν) caused exclusively by water molecules. In the infrared range, well-known internal modes ν1, ν2, and ν3 of the H2O molecule are observed for both polarizations, indicating the presence of water-I and water-II molecules in the crystal. Spectra recorded below 1000 cm(-1) reveal a rich set of highly anisotropic features in the low-energy response of H2O molecule in a crystalline nano-cavity. While for E∥c only two absorption peaks are detected, at ~90 cm(-1) and ~160 cm(-1), several absorption bands are discovered for E⊥c, each consisting of narrower resonances. The bands are assigned to librational (400-500 cm(-1)) and translational (150-200 cm(-1)) vibrations of water-I molecule that is weakly coupled to the nano-cavity "walls." A model is presented that explains the "fine structure" of the bands by a splitting of the energy levels due to quantum tunneling between the minima in a six-well potential relief felt by a molecule within the cavity.


Inhibitors of indoleamine 2,3-dioxygenase: a review of novel patented lead compounds.
The enzyme indoleamine 2,3-dioxygenase (IDO) regulates immune responses through the capacity to degrade the essential amino-acid tryptophan into kynurenine and other downstream metabolites that suppress effector T-cell function and favour the differentiation of regulatory T cells. The current experimental evidence indicates that IDO can be expressed by a variety of cell types, including dendritic cells, tumour cells and stromal cells. Recently, IDO has been implicated in B-cell stimulation and autoantibody production in experimental models of autoimmune diseases. Advances in the biochemistry of IDO and our understanding of the biological relevance of IDO-mediated tryptophan consumption to the establishment of immune tolerance are summarised and discussed. A selection of recent patents in the field are also reviewed and analysed. Readers will gain an overview of the patented compounds with IDO inhibitory activity from an immunologist's perspective. They will also learn about the companies that are main players in the field. Current evidence points to IDO as a molecular target for therapeutic intervention in order to restrain unwanted inflammatory/autoimmune responses and/or to boost antitumour immunity.


Alternative bearing surfaces in total hip arthroplasty.
Polyethylene wear and extension of indications of total hip arthroplasty into younger and younger age groups have pushed manufacturers to develop more durable bearing surfaces. Standard polyethylene, the plastic used for the first 3 decades of hip replacement, virtually ceases to exist in its original form. Modifications of the processing, including sterlization in an inert environment and cross-linking, have demonstrated some improvements in wear. Hard-on-hard bearings such as ceramic-on-ceramic and metal-on-metal also have demonstrated extremely low wear. This article reviews the pros and cons of the alternative bearing options available to assist in the proper bearing selection for a particular patient.


Effects of vanadate on hepatic cytochrome P-450 expression in streptozotocin-diabetic rats.
The ability of sodium metavanadate to reverse the effects of streptozotocin-induced diabetes on hepatic cytochrome P-450 isozymes was examined in male rats. Streptozotocin caused P-450h levels to fall 95%, and P-450j and P-450b levels to rise 8- and 40-fold, respectively, after 1 week. When diabetic rats were administered metavanadate in the drinking water for 7 days, P-450h apoprotein and mRNA levels remained no different from those of untreated diabetic rats, whereas levels of P-450b and P-450j decreased toward those of control rats. Metavanadate also lowered serum triglyceride and 3-hydroxybutyrate levels without lowering serum glucose in the diabetic rats. Furthermore, P-450h mRNA levels correlated well with levels of P-450h apoprotein for all treatment groups, indicating that P-450h suppression in diabetic rats is under pretranslational control and is independent of the increased expressions of P-450j and P-450b, and of the hyperlipidemia and ketosis that occurs in diabetes. Vanadate is capable of separating the effects of diabetes on expression of individual P-450 isozymes.


Directional Dicke Subradiance with Nonclassical and Classical Light Sources.
We investigate Dicke subradiance of N≥2 distant quantum sources in free space, i.e., the spatial emission patterns of spontaneously radiating noninteracting multilevel atoms or multiphoton sources, prepared in totally antisymmetric states. We find that the radiated intensity is marked by a full suppression of spontaneous emission in particular directions. In resemblance to the analogous, yet inverted, superradiant emission profiles of N distant two-level atoms prepared in symmetric Dicke states, we call the corresponding emission patterns directional Dicke subradiance. We further derive that higher-order intensity correlations of the light emitted by statistically independent thermal light sources display the same directional Dicke subradiant behavior and show that it stems from the same interference phenomenon as in the case of quantum sources. We finally present measurements of directional Dicke subradiance for N=2,…,5 distant thermal light sources corroborating the theoretical findings.


Cerebral: a Cytoscape plugin for layout of and interaction with biological networks using subcellular localization annotation.
Cerebral (Cell Region-Based Rendering And Layout) is an open-source Java plugin for the Cytoscape biomolecular interaction viewer. Given an interaction network and subcellular localization annotation, Cerebral automatically generates a view of the network in the style of traditional pathway diagrams, providing an intuitive interface for the exploration of a biological pathway or system. The molecules are separated into layers according to their subcellular localization. Potential products or outcomes of the pathway can be shown at the bottom of the view, clustered according to any molecular attribute data-protein function-for example. Cerebral scales well to networks containing thousands of nodes. http://www.pathogenomics.ca/cerebral


The Histone Demethylase KDM3A, Increased in Human Pancreatic Tumors, Regulates Expression of DCLK1 and Promotes Tumorigenesis in Mice.
The histone lysine demethylase 3A (KDM3A) demethylates H3K9me1 and H3K9Me2 to increase gene transcription and is upregulated in tumors, including pancreatic tumors. We investigated its activities in pancreatic cancer cell lines and its regulation of the gene encoding doublecortin calmodulin-like kinase 1 (DCLK1), a marker of cancer stem cells. We knocked down KDM3A in MiaPaCa-2 and S2-007 pancreatic cancer cell lines and overexpressed KDM3A in HPNE cells (human noncancerous pancreatic ductal cell line); we evaluated cell migration, invasion, and spheroid formation under hypoxic and normoxic conditions. Nude mice were given orthotopic injections of S2-007 cells, with or without (control) knockdown of KDM3A, and HPNE cells, with or without (control) overexpression of KDM3A; tumor growth was assessed. We analyzed pancreatic tumor tissues from mice and pancreatic cancer cell lines by immunohistochemistry and immunoblotting. We performed RNA-sequencing analysis of MiaPaCa-2 and S2-007 cells with knockdown of KDM3A and evaluated localization of DCLK1 and KDM3A by immunofluorescence. We analyzed the cancer genome atlas for levels of KDM3A and DCLK1 messenger RNA in human pancreatic ductal adenocarcinoma (PDAC) tissues and association with patient survival time. Levels of KDM3A were increased in human pancreatic tumor tissues and cell lines, compared with adjacent nontumor pancreatic tissues, such as islet and acinar cells. Knockdown of KDM3A in S2-007 cells significantly reduced colony formation, invasion, migration, and spheroid formation, compared with control cells, and slowed growth of orthotopic tumors in mice. We identified KDM3A-binding sites in the DCLK1 promoter; S2-007 cells with knockdown of KDM3A had reduced levels of DCLK1. HPNE cells that overexpressed KDM3A formed foci and spheres in culture and formed tumors and metastases in mice, whereas control HPNE cells did not. Hypoxia induced sphere formation and increased levels of KDM3A in S2-007 cells and in HPNE cells that overexpressed DCLK1, but not control HPNE cells. Levels of KDM3A and DCLK1 messenger RNA were higher in human PDAC than nontumor pancreatic tissues and correlated with shorter survival times of patients. We found human PDAC samples and pancreatic cancer cell lines to overexpress KDM3A. KDM3A increases expression of DCLK1, and levels of both proteins are increased in human PDAC samples. Knockdown of KDM3A in pancreatic cancer cell lines reduced their invasive and sphere-forming activities in culture and formation of orthotopic tumors in mice. Hypoxia increased expression of KDM3A in pancreatic cancer cells. Strategies to disrupt this pathway might be developed for treatment of pancreatic cancer.


Engineering a human bone marrow model: a case study on ex vivo erythropoiesis.
Bone marrow, with its intricate, three-dimensional tissue structure facilitating cell-cell interactions, provides a microenvironment supporting the production of hundreds of billions of multilineal blood cells everyday. We have developed a three-dimensional bone marrow culture system in which marrow cells are cultured in a reactor packed with porous microspheres. The culture supports a three-dimensional growth configuration and multilineal hemopoiesis mimicking the bone marrow in vivo. We studied ex vivo human erythropoiesis using the three-dimensional culture system. The system sustained extensive erythropoiesis at low erythropoietin concentrations (0.2 U/mL), plus stem cell factor, interleukin-3, granulocyte-macrophage colony-stimulating factor, and insulin-like growth factor-I. Erythroid cell production lasted for more than 5 weeks, and the percentage of erythroid cells in the nonadherent cell population was approximately 60%. Flow cytometric analysis using cell surface markers specific for erythroid cells (CD71 and glycophorin-A) indicated that the culture produced early, intermediate, and late erythroid cells. As the culture progressed, the erythroid cell population shifted gradually toward mature cell types. When compared to the three-dimensional culture, the traditional flask cultures failed to support extensive erythropoiesis under the same conditions. This indicates that the three-dimensional bone marrow culture system provides a microenvironment conducive to erythropoiesis under more physiological conditions and is a better bone marrow model.


A calibration of cellulose isotopes in modern prostrate Nothofagus and its application to fossil material from Antarctica.
Carbon and oxygen isotopes (δ13C and δ18O) in tree rings are widely used to reconstruct palaeoclimate variables such as temperature during the Holocene (12 thousand years ago - present), and are used increasingly in deeper time. However, their use is largely restricted to arboreal trees, which excludes potentially important data from prostrate trees and shrubs, which grow in high latitude and altitude end-member environments. Here, we calibrate the use of δ13C and δ18O as climatic archives in two modern species of southern beech (Nothofagus) from Tierra del Fuego, Chile, at the southern limit of their current range. We show that prostrate trees are potentially suitable archives for recording climatological means over longer periods (on the order of decades), which opens up these important environments for tree ring isotope analysis. We then apply our new understanding to a remarkable late Neogene (17-2.5 Ma) fossil Nothofagus assemblage from the Transantarctic Mountains, Antarctica, representative of a prostrate tundra shrub growing during a period of significant ice sheet retreat. The δ13C of the fossil cellulose was found to be ~4‰ enriched relative to that of the modern tress. This is likely to be due to a combination of a more positive δ13C of contemporaneous atmospheric CO2 and enhanced water use efficiency at the fossil site. Using the cellulose-δ18O in the fossil wood, we are able to reconstruct precipitation oxygen isotopes over the Antarctic interior for the first time for this time period. The results show that δ18Oprecip over Antarctica was -16.0 ± 4.2‰, around 12‰ enriched relative to today, suggesting changes in the hydrological cycle linked to warmer temperatures and a smaller ice sheet.


Facilitating validation of prediction models: a comparison of manual and semi-automated validation using registry-based data of breast cancer patients in the Netherlands.
Clinical prediction models are not routinely validated. To facilitate validation procedures, the online Evidencio platform ( https://www.evidencio.com ) has developed a tool partly automating this process. This study aims to determine whether semi-automated validation can reliably substitute manual validation. Four different models used in breast cancer care were selected: CancerMath, INFLUENCE, Predicted Probability of Axillary Metastasis, and PREDICT v.2.0. Data were obtained from the Netherlands Cancer Registry according to the inclusion criteria of the original development population. Calibration (intercepts and slopes) and discrimination (area under the curve (AUC)) were compared between semi-automated and manual validation. Differences between intercepts and slopes of all models using semi-automated validation ranged from 0 to 0.03 from manual validation, which was not clinically relevant. AUCs were identical for both validation methods. This easy to use semi-automated validation option is a good substitute for manual validation and might increase the number of validations of prediction models used in clinical practice. In addition, the validation tool was considered to be user-friendly and to save a lot of time compared to manual validation. Semi-automated validation will contribute to more accurate outcome predictions and treatment recommendations in the target population.


Order parameters for the multistep crystallization of clathrate hydrates.
Recent reports indicate that the crystallization of clathrate hydrates occurs in multiple steps that involve amorphous intermediates and metastable clathrate crystals. The elucidation of the reaction coordinate for clathrate crystallization requires the use of order parameters able to identify the reactants, products, and intermediates in the crystallization pathway. Nevertheless, existing order parameters cannot distinguish between amorphous and crystalline clathrates or between different clathrate crystals. In this work, we present the first set of order parameters that discern between the sI and sII clathrate crystals, the amorphous clathrates, the blob of solvent-separated guests and the liquid solution. These order parameters can be used to monitor the advance of the crystallization and for the efficient implementation of methods to sample the rare clathrate nucleation events in molecular simulations. We illustrate the use of these order parameters in the analysis of the growth and the dissolution of clathrate crystals and the spontaneous nucleation and growth of clathrates under conditions of high supercooling.


TRIENNIAL GROWTH AND DEVELOPMENT SYMPOSIUM: Intramuscular fat deposition in ruminants and pigs: A transcriptomics perspective.
The genomics era has led to an explosion in the study of gene expression in production animals. Intramuscular fat (IMF) content (both high and low) and composition are major quality attributes of meat, and more than 90 transcriptomic studies of IMF deposition have been undertaken in the ruminants and pigs since 2001, with the majority since 2008. The studies have implicated many genes involved in the control of adipogenesis, lipogenesis, and deposition of IMF, but there is relatively little consistency between the different studies. However, the genes encoding the synthesis enzymes acetyl-CoA carboxylase α, fatty acid synthase, and stearoyl-CoA desaturase; the fatty acid binding protein 4; the potential signaling protein thyroid hormone responsive; and the regulators C/EBPα, PPARγ, and sterol regulatory element binding transcription factor 1 are supported by 5 or more of the 90 studies. By combining the results of all the studies, complete pathways for long-chain fatty acid (LCFA) and triacylglyceride (TAG) synthesis are identified, as are a number of genes encoding proteins probably associated with the storage of TAG and genes encoding a number of known and potential adipokines. In contrast, support for the association of lipolytic pathways with IMF percentage is less strong. Differences in experimental design-in particular, the age of the animals, the rate of IMF deposition at sampling, the past nutritional history of the animals used, and the complexities of using a tissue with mixed cell types-have contributed to the differences in results and interpretation. Biomarkers predictive of future IMF percentage, facilitating reaching optimal IMF content at slaughter, may have industry utility, but to be useful in animal biopsy and postslaughter samples, where multiple cell types are present, genes must be carefully chosen to ensure that they are informative about the expected processes. Despite these problems, candidate biomarkers for estimation of de novo intramuscular adipocyte LCFA synthesis, LCFA uptake rate by intramuscular adipocytes, and IMF deposition rate have been identified and examples of their utility have been published. However, further work is required to demonstrate how best to apply the assays for the benefit of the relevant livestock production industries.


Coral thermal tolerance: tuning gene expression to resist thermal stress.
The acclimatization capacity of corals is a critical consideration in the persistence of coral reefs under stresses imposed by global climate change. The stress history of corals plays a role in subsequent response to heat stress, but the transcriptomic changes associated with these plastic changes have not been previously explored. In order to identify host transcriptomic changes associated with acquired thermal tolerance in the scleractinian coral Acropora millepora, corals preconditioned to a sub-lethal temperature of 3°C below bleaching threshold temperature were compared to both non-preconditioned corals and untreated controls using a cDNA microarray platform. After eight days of hyperthermal challenge, conditions under which non-preconditioned corals bleached and preconditioned corals (thermal-tolerant) maintained Symbiodinium density, a clear differentiation in the transcriptional profiles was revealed among the condition examined. Among these changes, nine differentially expressed genes separated preconditioned corals from non-preconditioned corals, with 42 genes differentially expressed between control and preconditioned treatments, and 70 genes between non-preconditioned corals and controls. Differentially expressed genes included components of an apoptotic signaling cascade, which suggest the inhibition of apoptosis in preconditioned corals. Additionally, lectins and genes involved in response to oxidative stress were also detected. One dominant pattern was the apparent tuning of gene expression observed between preconditioned and non-preconditioned treatments; that is, differences in expression magnitude were more apparent than differences in the identity of genes differentially expressed. Our work revealed a transcriptomic signature underlying the tolerance associated with coral thermal history, and suggests that understanding the molecular mechanisms behind physiological acclimatization would be critical for the modeling of reefs in impending climate change scenarios.


Nedd8 hydrolysis by UCH proteases in Plasmodium parasites.
Plasmodium parasites are the causative agents of malaria, a disease with wide public health repercussions. Increasing drug resistance and the absence of a vaccine make finding new chemotherapeutic strategies imperative. Components of the ubiquitin and ubiquitin-like pathways have garnered increased attention as novel targets given their necessity to parasite survival. Understanding how these pathways are regulated in Plasmodium and identifying differences to the host is paramount to selectively interfering with parasites. Here, we focus on Nedd8 modification in Plasmodium falciparum, given its central role to cell division and DNA repair, processes critical to Plasmodium parasites given their unusual cell cycle and requirement for refined repair mechanisms. By applying a functional chemical approach, we show that deNeddylation is controlled by a different set of enzymes in the parasite versus the human host. We elucidate the molecular determinants of the unusual dual ubiquitin/Nedd8 recognition by the essential PfUCH37 enzyme and, through parasite transgenics and drug assays, determine that only its ubiquitin activity is critical to parasite survival. Our experiments reveal interesting evolutionary differences in how neddylation is controlled in higher versus lower eukaryotes, and highlight the Nedd8 pathway as worthy of further exploration for therapeutic targeting in antimalarial drug design.


Dietary fats altered nephrotoxicity profile of methylmercury in rats.
Weanling male Sprague-Dawley rats were administered semi-purified isocaloric diet containing soy oil (SO), seal oil (SE), docosahexaenoic acid (DHA), fish oil (FO) or lard (LA) for 28 days, and then gavaged with 0, 1 or 3 mg MeHg kg(-1) body weight per day and fed the same diet for 14 days. Serum and 24 h urine samples were collected on the day of necropsy, and analyzed for markers of kidney function and diseases. Kidney slices were analyzed for para-amino-hippurate (PAH) and tetraethylammonium (TEA) uptake, total mercury and MeHg content, and examined for pathological lesions. Total mercury and MeHg contents increased significantly and dose-dependently in all dietary groups. MeHg significantly increased relative kidney weight in all groups, serum creatinine in all except SO group, serum uric acid in the DHA and LA groups, serum Mg in all except the LA group, and urinary protein in the SO group. MeHg significantly decreased serum urea nitrogen in SE, FO and LA groups, urinary creatinine in the DHA group, PAH uptake in all except the SE group, and TEA uptake in all groups. MeHg caused nephrosis in all dietary groups. MeHg also significantly increased neutrophil counts in all except the SE group, decreased serum albumin and triglyceride in all except the DHA group, and increased serum total cholesterol in all groups, suggesting a nephrotic syndrome-like outcome. These results confirmed that kidney tubules are major targets of MeHg nephrotoxicity. Treatment with dietary fats did not prevent, but rather altered the profile of, nephrotoxicity of MeHg in rats.


Deletion of both the Tyrosine-Based Endocytosis Signal and the Endoplasmic Reticulum Retrieval Signal in the Cytoplasmic Tail of Spike Protein Attenuates Porcine Epidemic Diarrhea Virus in Pigs.
Porcine epidemic diarrhea virus (PEDV) causes high mortality in neonatal piglets. The PEDV spike (S) protein contains two intracellular sorting motifs, YxxΦ (tyrosine-based motif YEVF or YEAF) and KVHVQ at the cytoplasmic tail, yet their functions have not been fully elucidated. Some Vero cell-adapted and/or attenuated PEDV variants contain ablations in these two motifs. We hypothesized that these motifs contribute to viral pathogenicity. By transiently expressing PEDV S proteins with mutations in the motifs, we confirmed that the motif KVHVQ is involved in retention of the S proteins in the endoplasmic reticulum (ER)-Golgi intermediate compartment (ERGIC). In addition, we showed that the YxxΦ motif triggers endocytosis of S proteins. These two motifs synergistically regulate the level of S expressed on the cell surface. To investigate their role in viral pathogenicity, we generated three recombinant PEDVs by introducing deletions or a mutation in the two motifs of the infectious clone of PEDV PC22A strain (icPC22A): (i) icΔ10aa (ΔYxxΦEKVHVQ), (ii) icΔ5aa (ΔKVHVQ), and (iii) icYA (Y1378A, to an inactivated motif, AEVF). Infection of Vero cells with icΔ10aa resulted in larger syncytia and more virions, with reduced numbers of S protein projections on the surface compared with icPC22A. Furthermore, we orally inoculated five groups of 5-day-old gnotobiotic piglets with the three mutants, icPC22A, or a mock treatment. Mutant icΔ10aa caused less severe diarrhea rate and significantly milder intestinal lesions than icPC22A, icΔ5aa, and icYA. These data suggest that the deletion of both motifs can reduce the virulence of PEDV in piglets.IMPORTANCE Many coronaviruses (CoVs) possess conserved motifs YxxΦ and/or KxHxx/KKxx in the cytoplasmic tail of the S protein. The KxHxx/KKxx motif has been identified as the ER retrieval signal, but the function of the YxxΦ motif in the intracellular sorting of CoV S proteins remains controversial. In this study, we showed that the YxxΦ of PEDV S protein is an endocytosis signal. Furthermore, using reverse genetics technology, we evaluated its role in PEDV pathogenicity in neonatal piglets. Our results explain one attenuation mechanism of Vero cell-adapted PEDV variants lacking functional YxxΦ and KVHVQ motifs. Knowledge from this study may aid in the design of efficacious live attenuated vaccines against PEDV, as well as other CoVs bearing the same motif in their S protein.


Interleukin-6 receptor blockade selectively reduces IL-21 production by CD4 T cells and IgG4 autoantibodies in rheumatoid arthritis.
Interleukin-6 (IL-6) levels are known to be increased in patients with rheumatoid arthritis (RA). Tocilizumab, a monoclonal antibody to the IL-6 receptor (IL-6R), reduces disease activity in RA, although its mechanisms of action remain unclear. Since IL-6 regulates cytokine production by CD4 T cells during activation, we investigated whether treatment with tocilizumab altered the phenotype and cytokine production by CD4 T cells in patients with rheumatoid arthritis. We show here that tocilizumab treatment does not change the production of cytokines by naïve CD4 T cells. However, tocilizumab treatment causes a selective decrease of IL-21 production by memory/activated CD4 T cells. Since IL-21 is known to promote plasma cell differentiation, we examined the effect of tocilizumab on the production of autoantibodies. We show that there is a decrease in the levels of IgG4 anti-CCP antibodies, but there is no effect on IgG1 anti-CCP antibodies. In addition, we show that IL-21 is a powerful inducer of IgG4 production by B cells. Thus, IL-6 contributes to the presence of IgG4-specific anti-CCP autoantibodies in RA patients, likely through its effect on IL-21 production by CD4 T cells, and IL-6R blockade down-regulates this pathway.


Comparison of the risk of malignancy index and self-constructed logistic regression models in preoperative evaluation of adnexal masses.
The aim of this study was to evaluate women with adnexal masses in the preoperative period by creating 2 logistic regression models, 1 including sonographic morphologic characteristics and the other including both morphologic and color Doppler characteristics, to compare the diagnostic accuracy of these 2 models with the risk of malignancy index (RMI). This prospective study included 38 malignant, 7 borderline, and 244 benign ovarian masses. The menopausal status, presence of septa, presence of papillary projections, location of the tumor, presence of ascites, presence of metastases, cancer antigen 125 level, tumor volume, septa thickness, and percentage of the solid component were included in the initial analysis. A second regression analysis was performed with the addition of Doppler parameters (location of blood flow and lowest resistive index) in the data set. Diagnostic performance of the 2 regression models and RMI were described and compared by generating receiver operating characteristic curves for each model. The area under the curve values for the morphologic model (model 1), Doppler model (model 2), and RMI were 0.907, 0.971, and 0.889, respectively. Significance levels of model 1 and the RMI were similar (P = .23), whereas model 2 had a significantly higher area under the curve compared with both model 1 (P = .037) and the RMI (P = .018). The addition of Doppler parameters in the regression model significantly increases the predictive performance. Nevertheless, in low-resource settings, the RMI remains the method of choice for distinguishing adnexal masses and referral to gynecologic oncology clinics.


International efforts in geriatric radiation oncology.
Geriatric assessment (GA) has been recommended to form part of treatment decision making for older adults with cancer. However despite consensus guidelines from various organizations, GA does not appear to be a part of routine practice in radiation oncology. The aim of the current study was to explore the implementation of GA in radiation oncology. This anonymous international survey investigated current use of GA in patients presenting for radiation therapy aged 65 years and over, in accordance with Checklist for Reporting Results of Internet E-Surveys (CHERRIES) guidelines. The survey was designed, using Qualitrics™, an online survey tool. It was distributed via SIOG, social media and radiation oncology professional organizations. Survey responses were analyzed using simple descriptive statistics. An additional analysis by creating a dichotomous variable based on awareness of major clinical practice guidelines and current use of GA. Among 158 respondents, there was relatively low awareness of GA guidelines and low uptake of validated tools and processes. A minority of participants, only 16%, stated that they had a specialized geriatric oncology program in their institution. Approximately a third (34%) of respondents were unaware of any GA clinical practice guidelines. With regard to what way participants assess older patients differently to younger patients, 16% reported formally using specific validated tools, whereas 73% reported an informal assessment based on their own judgment, with 5% reporting no difference between younger and older patients. Regarding the use of validated screening tools for geriatric impairments, over half reported using none (57%). Regarding GA implementation, the main barriers highlighted included a lack of clinical/support staff, a lack of training, knowledge, understanding or experience about GA and a lack of time. Relatively low awareness of guidelines and low uptake of formal GA tools and processes were found. The integration of GA principles into radiation oncology appears to be ad hoc and very much in its infancy. There is a clear need for increased interdisciplinary education and collaboration between the disciplines of radiation oncology and geriatric medicine.


Iodine isotopes in radiation hygiene.
Radioactive isotopes of iodine rank among the factors of anthropogenic contamination of the biosphere. The review presents a radiation-hygienic survey of literary data and the author's own results of studies of the metabolism and biological effect of radioactive isotopes of iodine. The risk of their intake by human organism is evaluated. Particular attention is paid to 132-135I and 129I. Data on biological danger of these isotopes are scarce in the literature.


Loss of nuclear pro-IL-16 facilitates cell cycle progression in human cutaneous T cell lymphoma.
Cutaneous T cell lymphomas (CTCLs) represent a heterogeneous group of non-Hodgkin lymphomas that affect the skin. The pathogenesis of these conditions is poorly understood. For example, the signaling mechanisms contributing to the dysregulated growth of the neoplastic T cells are not well defined. Here, we demonstrate that loss of nuclear localization of pro-IL-16 facilitates CTCL cell proliferation by causing a decrease in expression of the cyclin dependent-kinase inhibitor p27Kip1. The decrease in p27Kip1 expression was directly attributable to an increase in expression of S-phase kinase-associated protein 2 (Skp2). Regulation of Skp2 is in part attributed to the nuclear presence of the scaffold protein pro-IL-16. T cells isolated from 11 patients with advanced CTCL, but not those from healthy controls or patients with T cell acute lymphocytic leukemia (T-ALL), demonstrated reduction in nuclear pro-IL-16 levels. Sequence analysis identified the presence of mutations in the 5' end of the PDZ1 region of pro-IL-16, a domain required for association of pro-IL-16 with the nuclear chaperone HSC70 (also known as HSPA8). HSC70 knockdown led to loss of nuclear translocation by pro-IL-16 and subsequent increases in Skp2 levels and decreases in p27Kip1 levels, which ultimately enhanced T cell proliferation. Thus, our data indicate that advanced CTCL cell growth is facilitated, at least in part, by mutations in the scaffold protein pro-IL-16, which directly regulates Skp2 synthesis.


Dermatoglyphs of digito-palmar complex in autistic disorder: family analysis.
To examine the role of the genetic component in the quantitative dermatoglyphic traits of autistic patients and their families, and the transmission of the autism. Finger and palm prints were taken from 120 autistic patients (92 males and 28 females), their parents (92 mothers and 70 fathers), 32 healthy brothers and 28 sisters, as well as 400 healthy controls (200 males and 200 females). An analysis of quantitative traits of dermatoglyphs on the fingers (FRC - finger ridge count) and palms (a-b, b-c, and c-d ridge count, and atd angle) was performed. Descriptive statistics, multivariate analysis of variance (MANOVA) with Tukey HSD post hoc test, and discriminant analysis were used to determine the differences among the groups. In addition, correlations among family members were analyzed. Multivariate analysis showed significant differences among examined groups of autistic patients and their family members and healthy volunteers regarding both group membership and sex. Autistic male patients differed significantly from the healthy controls in the ridge count (RC) on the fourth and fifth finger, and in a-b RC and atd angle of both hands. Healthy fathers of autistic patients differed in atd angle, and brothers of autistic patients differed in all palmar variables from the healthy control group. Mothers of autistic patients differed significantly from the healthy female controls in the RC of the first, fourth, and fifth finger, in a-b and c-d RC on the palms, and atd angle of both hands. The first two discriminant functions explained 85.4% of variance and separated groups clearly in two ways: the first function separated healthy controls from family members of autistic patients, and the second one males from females. Interfamilial analysis showed significant interclass correlations between autistic sons and their mothers or fathers in practically all variables. However, the correlation between parents and their autistic daughters was lower. Both mothers and fathers of autistic patients correlated with their healthy children only in palmar variables. We found significant differences in ridge counts on the fingers and palms between the affected patients and their healthy controls, but these differences also existed between family members of autistic patients and healthy controls. Particularly pronounced were the differences between healthy female controls and female family members, including not only autistic female patients, but also their healthy mothers and sisters. Since the mothers and their autistic sons showed higher statistically significant correlation in most of the examined variables, unlike the mothers and their autistic or healthy daughters, it is possible that there is a connection between a recessive X-chromosome linkage, as a genetic component in the etiology of autistic disorders, and the influence of the inactivation of the affected X-chromosome in the females.


Massive localised lymphoedema: a rare vascular malformation.
Lymphatic malformations are a subset of congenital vascular malformations, and are caused by a defect in lymphatic development during embryogenesis. When lymphatic mesoderm development is prematurely arrested, it retains it proliferative potential. Stimulus in the future can cause the lesion to proliferate locally without coordination or regulation, resulting in the rare condition known as massive localised lymphoedema (MLL). We present a case report of MLL, a rare and ill-defined soft tissue mass reported in the morbidly obese, with reference to the existing literature.


gamma-Aminobutyric acid inhibits beta-endorphin secretion from the anterior pituitary but not the neurointermediate lobe in the rat.
We have evaluated the role of gamma-aminobutyric acid (GABA) in the neuroendocrine control of beta-endorphin (beta-EP) secretion in the rat. Plasma beta-EP and beta-lipotropin (beta-LPH) levels and beta-EP-like immunoreactivity (beta-EPLI) in the anterior pituitary (AP) and neurointermediate lobe (NIL) were determined after administration of GABA antagonist or agonist drugs in male rats under resting conditions or after potent physical stresses. Bicuculline (0.1-0.8 mg/kg BW ip), a GABA receptor antagonist, induced a dose-related rise in plasma beta-EP and beta-LPH levels and a concomitant decrease in beta-EPLI concentrations in the AP but not in the NIL. Muscimol, a potent GABA-mimetic drug, did not alter baseline plasma beta-EP and beta-LPH levels, whether given systemically (1.0-2.0 mg/kg BW ip) or intracerebroventricularly (500 ng/kg BW), but prevented the effect of bicuculline on plasma and AP-beta-EP and beta-LPH concentrations. Administration of foot shock or restraint stress induced a clear-cut activation of the AP-related beta-EP secretion, an effect that was prevented by pretreatment with muscimol. Together, these data show that GABA-ergic mechanisms, probably operating at a central nervous system level, exert an inhibitory action on resting and stimulated beta-EP and beta-LPH secretion. Since no alterations in beta-EP concentrations in the NIL occurred after manipulations with GABA-ergic drugs or stress, and these were detected only in the AP, an interaction between GABA-ergic neurons and CRF neurons is the most likely explanation for the reported findings.


Pre-transplant donor-specific transfusions induce allograft rejection and IL-2 gene expression in the WKY-->F344 functional tolerance model of rat lung transplantation.
Our previous studies have shown that a spontaneous functional tolerance develops in a rat model of lung transplantation (WKY-->F344). The tolerance observed in this model may be due to the minor histocompatible differences in this combination, however, the possibility of a tolerogenic effect related specifically to the lung allograft must be considered. To further examine this model, the effect of pre-transplant donor-specific spleen cell transfusions (DSTs) was examined on the functional tolerance state seen in this model. F344 rats received WKY spleen cells on days -45 and -30 before lung transplantations. Control F344 rats received lung transplants without DSTs. Recipients in both groups were killed on day 7, 14, 21 and 49 post-transplant, and allograft rejection (AR) was graded histologically (stage 0-IV). Intragraft cytokine gene transcripts were examined on day 7 and 14 post-transplant using reverse transcriptase-polymerase chain reaction (RT-PCR) techniques to investigate the underlying immunological events occurring in each group. In addition, allogeneic (WKY) and third party (BN) skin grafts were placed on lung recipients at day 35 post-transplant to evaluate the development of systemic tolerance. It was seen that control animals showed moderate to severe lymphocytic infiltrations (stage II-III AR) in the first 3 weeks followed by spontaneous recovery with stage I-II AR on day 49. In marked contrast, DST-treated animals showed more aggressive AR with severe lymphocytic infiltration and haemorrhagic infarction (stage III-IV AR) by day 14-21, without any evidence of recovery on day 49. WKY skin grafts showed prolonged survival in control animals, but were promptly rejected in DST-treated animals. Intragraft cytokine gene expression in control animals was characterized by no or weak expression of IL-2 and high IL-10, while DST-treated animals showed high levels of IL-2 transcripts. IL-2:IL-10 and IL-2:IL-4 ratios were significantly increased in DST-treated animals compared with controls on day 7 post-transplant. It was concluded that pre-transplant DSTs did not enhance allograft survival, but actually induced AR and ablated any immunological benefit of the lung allograft on induction of tolerance in the WKY-->F344 lung transplant model. It was found that the DST-induced AR was associated with a deviation of cytokine immune responses from a predominant Th2 to Th1 profile characterized by increased IL-2 gene expression in the allografts. We also conclude that factors other than the degree of histocompatibility matching, such as the route and timing of alloantigen exposure, and the amount or nature of alloantigens associated specifically with lung allografts, are involved in deviating native immune responses toward acceptance or rejection of lung allografts in this model of lung transplantation.


Comparative study of single-, double-, and triple-nerve transfer to a common target: experimental study of rat brachial plexus.
The purpose of this study was to investigate the recovery of a common target motor function after different single and combined motor nerve transfers in rat brachial plexus model. The musculocutaneous nerve and biceps muscle were chosen as the target for neurotization. The phrenic, pectoral, and suprascapular nerves were selected as the neurotizers. Forty-two Sprague-Dawley rats were randomly assigned to seven groups (six rats in each group): single-neurotizer transfer (three groups), double-neurotizer transfer (three groups), and triple-neurotizer transfer (one group). The contralateral intact forelimb was used as a control. Functional outcomes were measured by grooming test, electrophysiological study, muscle contraction strength, muscle weight, and axon counts. At 12 weeks, 40 operative rats were studied (two had died). In the single-neurotizer transfer, all three transfers showed no significant difference in motor recovery of the biceps. In the double-neurotizer transfer groups, the worst results were seen in group 6 (combined pectoral and suprascapular nerve transfer) despite increasing axon counts. This study may potentially suggest: (1) single-neurotizer transfer will not have synergistic or antagonistic effects; (2) two neurotizers with functional antagonism will significantly downgrade motor recovery of the neurotized muscle despite increasing axon counts; (3) multiple motor neurotizer transfers may not always provide a better outcome, although increasing axons may outweigh antagonistic effects; and (4) phrenic nerve transfer alone did not upgrade the functional outcome despite its automatic discharge. Any nerve transfer combined with phrenic nerve transfer, however, showed improved functional results.


[Mechanism of metabolic adaptation].
The paper presents results of investigations on some mechanisms of metabolic adaptation in mammals. There are four sections in the tissue metabolic system of acid-base homeostasis: polyamines as factors of metabolic adaptation; significance of carbon dioxide for metabolic response formation in hypobiosis; polyamines metabolism in hypobiosis. Peculiarities of intermediate metabolism have been analyzed in animal tissues under the changes in H+, CO2 and HCO3- concentrations. Basing on a new interpretation of the experimental data and detected regularity in the metabolism, conclusion on the existence of a new acid-base homeostasis system in the tissues has been made. The results of polyamines metabolism investigations in the mammals under the stress have been described. The experimental data make us to believe that changes in polyamines synthesis and ODC activity in particular, is a part of stereotype nonspecific response to any stress impacts and one of the factors of cell metabolic adaptation. Some new data on mechanisms of formation and control of metabolic status of animals in the natural and artificial hibernation have been presented. The key idea is that in the state of hypobiosis the carbon dioxide (HCO3-) appears as a regulatory factor of metabolic adaptation, which is able to realize its action directly via affecting numerous biochemical events. The participation of polyamines in adaptive metabolic response to hybernation factors is suggested. Some peculiarities of ornithine decarboxilase and transglutaminase activity during the different stages of genuine and artificial hypobiosis have been demonstrated.


Intake of garlic and its bioactive components.
The health benefits of garlic likely arise from a wide variety of components, possibly working synergistically. The complex chemistry of garlic makes it plausible that variations in processing can yield quite different preparations. Highly unstable thiosulfinates, such as allicin, disappear during processing and are quickly transformed into a variety of organosulfur components. The efficacy and safety of these preparations in preparing dietary supplements based on garlic are also contingent on the processing methods employed. Although there are many garlic supplements commercially available, they fall into one of four categories, i.e., dehydrated garlic powder, garlic oil, garlic oil macerate and aged garlic extract (AGE). Garlic and garlic supplements are consumed in many cultures for their hypolipidemic, antiplatelet and procirculatory effects. In addition to these proclaimed beneficial effects, some garlic preparations also appear to possess hepatoprotective, immune-enhancing, anticancer and chemopreventive activities. Some preparations appear to be antioxidative, whereas others may stimulate oxidation. These additional biological effects attributed to AGE may be due to compounds, such as S-allylcysteine, S-allylmercaptocysteine, N(alpha)-fructosyl arginine and others, formed during the extraction process. Although not all of the active ingredients are known, ample research suggests that several bioavailable components likely contribute to the observed beneficial effects of garlic.


Organization of NADPH-cytochrome P450 reductase and CYP1A2 in the endoplasmic reticulum--microdomain localization affects monooxygenase function.
Cytochrome P450 is part of an electron transport chain found in the endoplasmic reticulum (ER), with its catalytic function requiring interactions with NADPH-cytochrome P450 reductase (CPR). The goals of this study were to examine how the P450 system proteins are organized in the membrane and to determine whether they are distributed in detergent-resistant lipid microdomains (DRM). Isolated liver microsomes from untreated rabbits were treated with 1% Brij 98, and DRMs were isolated via sucrose gradient centrifugation. Lipid analysis showed that DRM fractions were enriched in cholesterol and sphingomyelin, similar to that found with plasma membrane DRMs. Approximately 73% of CYP1A2 and 68% of CPR resided in DRM fractions, compared with only 33% of total ER proteins. These DRMs were found to be cholesterol-dependent: CPR and CYP1A2 migrated to the more dense regions of the sucrose gradient after cholesterol depletion. CYP1A2 function was studied in three purified lipid vesicles consisting of 1) phosphatidylcholine (V-PC), 2) lipids with a composition similar to ER lipids (V-ER), and 3) lipids with a composition similar to the DRM fractions (V-DRM). Each system showed similar substrate binding characteristics. However, when the association between CPR and CYP1A2 was measured, V-ER and V-DRM liposomes produced lower apparent K(m) values compared with V-PC without any significant change in V(max). These findings suggest that CYP1A2 and CPR reside in ER-DRMs and that the unique lipid components of these domains enhance CYP1A2 substrate metabolism through greater efficiency in CPR-CYP1A2 binding.


Early Rehabilitation with Exergame - Which Muscle Groups are Addressed by Bedside-Based Knee Exercises?
Knee exercises are generally recommended following knee-related operations. Nevertheless, compliance in performing self-administered exercises is varying. The use of exergames may alter patient compliance. These computer games encourage the user to exercise. Exergames are not yet used much in orthopedic rehabilitation. The aim of this study was to analyze the influence of different seating positions in electromyography while performing knee exercises with the exergame GenuSport. 72 healthy women and men were tested in 6 seating positions using an application-based knee-training device (KT). Positions were 1. user seated 90 upright with straight leg on KT, 2. 90 upright with leg exercising in external rotation on KT, 3. 90 upright with leg exercising in internal rotation on KT, 4. 90 upright with straight leg on 30mm elevated KT, 5. 45 upright with straight leg on KT and 6. 45 upright with straight leg on 30mm elevated KT. Electromyogram was recorded for biceps femoris, rectus femoris, semitendinosus, vastus medialis, adductor magnus and vastus lateralis. No significant difference was shown in position 2, 3 or 4 compared to position 1. In position 5 and 6, EMG reported significantly higher values compared to positions 1 to 4 (ischiocrural: 130% and quadriceps: 90%). The quadriceps was activated significantly less in position 6 compared to 5. 1. Bedside-based knee extension exercises are most effective in a seated position of 45 trunk inclination. 2. A slightly more bent knee (KT elevation of 30mm) leads to a higher activation of the ischiocrural muscles.


Sepsis and septic shock.
For more than two decades, sepsis was defined as a microbial infection that produces fever (or hypothermia), tachycardia, tachypnoea and blood leukocyte changes. Sepsis is now increasingly being considered a dysregulated systemic inflammatory and immune response to microbial invasion that produces organ injury for which mortality rates are declining to 15-25%. Septic shock remains defined as sepsis with hyperlactataemia and concurrent hypotension requiring vasopressor therapy, with in-hospital mortality rates approaching 30-50%. With earlier recognition and more compliance to best practices, sepsis has become less of an immediate life-threatening disorder and more of a long-term chronic critical illness, often associated with prolonged inflammation, immune suppression, organ injury and lean tissue wasting. Furthermore, patients who survive sepsis have continuing risk of mortality after discharge, as well as long-term cognitive and functional deficits. Earlier recognition and improved implementation of best practices have reduced in-hospital mortality, but results from the use of immunomodulatory agents to date have been disappointing. Similarly, no biomarker can definitely diagnose sepsis or predict its clinical outcome. Because of its complexity, improvements in sepsis outcomes are likely to continue to be slow and incremental.


Reversal of Penicillin-Induced L-Phase Growth of Haemophilus influenzae by Spermine and Its Effects on Antibiotic Susceptibility.
Spermine at a concentration of 0.001 m initiated the reversion of penicillin-induced L-phase growth of Haemophilus influenzae to bacillary growth on penicillin L-phase medium. Reversion of L-phase colonies to bacillary colonies required 3 to 5 days. This spermine reversal of variant growth also occurred on penicillin induction medium in the presence of concentrations of tetracycline, erythromycin, and chloramphenicol that were not bactericidal for L-phase or bacillary inocula until 48 or more hr. Spermine was without protective effect against streptomycin and kanamycin, which were, in combination with penicillin, bactericidal at 24 hr. Spermine protection of L-phase variants against antibiotic toxicity was, therefore, related to initiation (by spermine) or bacillary growth from round bodies that survived for 24 or more hr at bacteriostatic levels of antibiotic.


Postconcussive symptoms and neurocognitive function after mild traumatic brain injury in children.
We describe children's postconcussive symptoms (PCSs), neurocognitive function, and recovery during 4 to 5 weeks after mild traumatic brain injury (MTBI) and compare performance and recovery with those of injured control group participants without MTBIs. A prospective, longitudinal, observational study was performed with a convenience sample from a tertiary care, pediatric emergency department. Participants were children 10 to 17 years of age who were treated in the emergency department and discharged. The MTBI group included patients with blunt head trauma, Glasgow Coma Scale scores of 13 to 15, loss of consciousness for < or = 30 minutes, posttraumatic amnesia of < or = 24 hours, altered mental status, or focal neurologic deficits, and no intracranial abnormalities. The control group included patients with injuries excluding the head. The Post-Concussion Symptom Questionnaire and domain-specific neurocognitive tests were completed at baseline and at 1 and 4 to 5 weeks after injury. Twenty-eight MTBI group participants and 45 control group participants were compared. There were no significant differences in demographic features. Control group participants reported some PCSs; however, MTBI group participants reported significantly more PCSs at all times. Among MTBI group participants, PCSs persisted for 5 weeks after injury, decreasing significantly between 1 and 4 to 5 weeks. Patterns of recovery on the Trail-Making Test Part B differed significantly between groups; performance on other neurocognitive measures did not differ. In children 10 to 17 years of age, self-reported PCSs were not exclusive to patients with MTBIs. However, PCSs and recovery patterns for the Trail-Making Test Part B differed significantly between the groups.


Mutagenicity of isomeric alkanediazotates, precursors for ultimate alkylating species of carcinogenic N-nitroso compounds.
Alkanediazohydroxides are common key intermediates in carcinogenesis and mutagenesis of N-nitroso compounds, which are widely found in human environment. Mutagenicity of (E)- and (Z)-potassium alkanediazotates, as precursors of corresponding alkanediazohydroxides were evaluated to investigate the effect of geometric isomerism and also the effect of alkyl groups on their biological activity. Mutagenicity of N-nitroso-N-alkylureas which spontaneously produce alkanediazohydroxides after non-enzymatic hydrolysis were also tested in comparison to that of the corresponding diazotates and other activated chemical species of N-nitrosamines. When the mutagenicity was assayed in three microbial strains, Salmonella typhimurium TA1535, and Escherichia coli WP2 and WP2 uvrA, the order of mutagenic potency of the compounds with the same alkyl group was as follows; (E)-diazotates > (Z)-diazotates > nitrosoureas. The effect of alkyl groups on the mutagenic potency was different in Salmonella strain and in E. coli strains, and this result could be explained by the efficiency of O6-alkylguanine-DNA alkyltransferase. In each bacterial strain, this effect of alkyl groups was similar in mutagenicity induced by (E)- and (Z)-diazotates, N-nitroso-N-alkylureas and other activated N-nitrosodialkylamines such as alpha-hydroxy nitrosamines. The geometrical isomerism affected the mutagenicity of (E)- and (Z)-potassium alkanediazotates, and the result suggested that alkanediazohydroxides react through diazonium ions in a cage rather than through free alkyldiazonium ions which have no geometrical isomerism. Our results confirmed that (E)-potassium alkanediazotates, (Z)-potassium alkanediazotates and N-nitroso-N-alkylureas all decomposed through diazohydroxides, and that alkanediazohydroxides are the active alkylating species of N-nitroso compounds, and also that the geometrical isomerism is important for carcinogenic N-nitroso compounds to show their biological activity.


[Modeling of rheographic curves for testing equipment].
The authors describe the method of obtaining model rheographic curves applied in testing equipment for appliances investigating cardiovascular system. The method is based on the electrical model of the left ventricle of the heart, aortic and mitral valves and vessels of the greater circulation. Relevant mathematical models are provided. Rheographic curves that were drawn in time by mathematical models are presented.


An evaluation of a modified preventive drenching programme on commercial sheep farms.
The effectiveness of a commonly advocated five drench preventive drenching programme in lambs was assessed on nine farms in the Gisborne, Hawkes Bay and Wairarapa districts between December 1983 and March 1985. The productive performance of two groups of preventively drenched lambs was compared with lambs in which parasitism was suppressed by fortnightly drenching from weaning until mid-August 1984. One of the groups received the standard five drench programme commencing at weaning and finishing in March 1984. The other group received six additional 4-weekly drenches, with the last drench in mid-August 1984. All the other lambs on the farms were drenched according to the five drench programme. The availability of infective larvae on pasture and the build-up of parasitic infection in the lambs was monitored. The effects of the different drenching programmes on liveweight and faecal egg counts was followed until March 1985, when the animals were 18 months old. Between March and July 1984 on all nine farms there was a significant depression of growth rates of the standard five drench group when compared with the suppression-drenched group (P<0.01). The mean liveweight difference from December 1983 to July 1984 for all farms was 4.0 kg (P<0.05). Hogget wool weights were significantly depressed (mean difference 0.37 kg, P<0.05). Faecal egg counts rose steadily after the completion of the five drench programme in March, exceeding 1000 eggs per gram on seven farms and 4000 eggs per gram on three of these farms. Trichostrongylus spp. was the major constituent of the winter parasite increase except on one farm when Cooperia spp. succeeded a previous Haemonchus dominance. Two farms had major build-ups of Haemonchus spp., one in early May, the other in June. Six farms salvage-drenched the standard five drench groups. The lambs in the standard drench groups on these farms recovered about half the bodyweight difference between August and March. The mean weight gains to July and wool weights of the modified drench programme groups were reduced slightly compared with the suppression-drenched group (mean liveweight difference 1.4 kg; mean wool weight difference 0.15 kg). No major rises in faecal egg counts occurred. Herbage larval counts exceeded 1000 larvae/kg at some stage on seven farms and, of these, the counts exceeded 4000 larvae/kg on three farms.


Co-development of alcohol use problems and antisocial peer affiliation from ages 11 to 34: selection, socialization and genetic and environmental influences.
Social context is an important factor in determining the developmental trajectory of alcohol use. We examined the co-development between alcohol use problems and antisocial peer affiliation. We also estimated the genetic and environmental influences on alcohol use problems, antisocial peer affiliation and their co-development over time. Longitudinal study using bivariate latent basis models with structured residuals (LBM-SR). A biometric model was then fitted to estimate the genetic and environmental influences on the growth factors and their covariances. The United States mid-west region. Members of the Minnesota Twin Family Study (MTFS), an ongoing, longitudinal study of 3762 (52% female) twins (1881 pairs). Alcohol use problems were assessed using a composite measure of average number of drinks per occasion in the past 12 months, maximum number of drinks in 24 hours and DSM-III-R symptoms of alcohol abuse and dependence. Antisocial peer affiliation was measured by self-report of the proportion of one's friends who exhibited types of antisocial behaviors. The LBM-SR model revealed that there was a large correlation between the growth factors for alcohol use problems and antisocial peer affiliation [r = 0.78, 95% confidence interval (CI) = 0.76, 0.80] and cross-lagged effects consistent with both selection and socialization effects. Additionally, antisocial peer affiliation in adolescence was associated with greater increases in alcohol use problems over time (r = 0.57, 95% CI = 0.54, 0.60). Genetic influences largely accounted for the association between antisocial peer affiliation in pre-adolescence and growth in alcohol use problems, while shared environmental influences accounted for the correlation between antisocial peer affiliation and alcohol use problems growth factors. Antisocial peer affiliation in adolescence appears to be a salient, genetically influenced risk factor for early alcohol use and increase in alcohol use from adolescence to young adulthood.


POFUT1-mediated O-fucosylation of glycoproteins expressed in the baculovirus Sf9 insect cell expression system.
The baculovirus-insect cell expression system allows addition of O-fucose to EGF-like domains of glycoproteins, following the action of the protein O-fucosyltransferase 1 named POFUT1. In this study, recombinant Spodoptera frugiperda POFUT1 from baculovirus-infected Sf9 cells was compared to recombinant Mus musculus POFUT1 produced by CHO cells. Contrary to recombinant murine POFUT1 carrying two hybrid and/or complex type N-glycans, Spodoptera frugiperda POFUT1 exhibited paucimannose N-glycans, at least on its highly evolutionary conserved across Metazoa NRT site. The abilities of both recombinant enzymes to add in vitro O -fucose to EGF-like domains of three different recombinant mammalian glycoproteins were then explored. In vitro POFUT1-mediated O-fucosylation experiments, followed by click chemistry and blot analyses, showed that Spodoptera frugiperda POFUT1 was able to add O-fucose to mouse NOTCH1 EGF-like 26 and WIF1 EGF-like 3 domains, similarly to the murine counterpart. As proved by mass spectrometry, full-length human WNT Inhibitor Factor 1 expressed by Sf9 cells was also modified with O-fucose. However, Spodoptera frugiperda POFUT1 was unable to modify the single EGF-like domain of mouse PAMR1 with O-fucose, contrary to murine POFUT1. Absence of orthologous proteins such as PAMR1 in insects may explain the enzyme's difficulty in adding O-fucose to a domain that it never encounters naturally.


ACR Appropriateness Criteria® Syncope.
Syncope and presyncope lead to well over one million emergency room visits in the United States each year. Elucidating the cause of syncope or presyncope, which are grouped together given similar etiologies and outcomes, can be exceedingly difficult given the diverse etiologies. This becomes more challenging as some causes, such as vasovagal syncope, are relatively innocuous while others, such as cardiac-related syncope, carry a significant increased risk of death. While the mainstay of syncope and presyncope assessment is a detailed history and physical examination, imaging can play a role in certain situations. In patients where a cardiovascular etiology is suspected based on the appropriate history, physical examination, and ECG findings, resting transthoracic echocardiography is usually considered appropriate for the initial imaging. While no imaging studies are considered usually appropriate when there is a low probability of cardiac or neurologic pathology, chest radiography may be appropriate in certain clinical situations. The American College of Radiology Appropriateness Criteria are evidence-based guidelines for specific clinical conditions that are reviewed annually by a multidisciplinary expert panel. The guideline development and revision include an extensive analysis of current medical literature from peer reviewed journals and the application of well-established methodologies (RAND/UCLA Appropriateness Method and Grading of Recommendations Assessment, Development, and Evaluation or GRADE) to rate the appropriateness of imaging and treatment procedures for specific clinical scenarios. In those instances where evidence is lacking or equivocal, expert opinion may supplement the available evidence to recommend imaging or treatment.


Paliperidone LAI and Aripiprazole LAI Plasma Level Monitoring in the Prophylaxis of Bipolar Disorder Type I with Manic Predominance.
The objective of this study was the evaluation of utility of plasma level monitoring in the clinical stabilizing efficacy and tolerability of paliperidone palmitate (PP) vs. aripiprazole monohydrate (AM) in bipolar disorder I (BD I) with manic predominance. Fifty-six outpatients of both sexes, age ranging from 18 to 65 years, affected by BD I with manic predominance, orally treated and stabilized after acute episode for at least 2 weeks with paliperidone or aripiprazole (n=31, paliperidone; n=25, aripiprazole) underwent a prospective observational study of switching to the corresponding long-acting injection (LAI) on the basis of clinical evaluation. The efficacy and tolerability of the 2 treatments were assessed by BPRS, PANSS, HAMD21, and MRS rating scales and a check list every month for 12 months. Drug plasma levels determinations (PLs) were performed at the same times. A good clinical stability and tolerability of both drugs were reported. Lower mean PLs of PP showed a positive effect on depressive symptoms. AM PLs variability was associated with greater instability of manic symptoms whereas intermediate PLs seem to have more influence on depressive symptomatology. PLs drug monitoring has been proven to be useful, and further investigations to identify optimal therapeutic ranges for LAI formulations are needed.


Factors influencing breath ammonia determination.
Amongst volatile compounds (VCs) present in exhaled breath, ammonia has held great promise and yet it has confounded researchers due to its inherent reactivity. Herein we have evaluated various factors in both breath instrumentation and the breath collection process in an effort to reduce variability. We found that the temperature of breath sampler and breath sensor, mouth rinse pH, and mode of breathing to be important factors. The influence of the rinses is heavily dependent upon the pH of the rinse. The basic rinse (pH 8.0) caused a mean increase of the ammonia concentration by 410 ± 221 ppb. The neutral rinse (pH 7.0), slightly acidic rinse (pH 5.8), and acidic rinse (pH 2.5) caused a mean decrease of the ammonia concentration by 498 ± 355 ppb, 527 ± 198 ppb, and 596 ± 385 ppb, respectively. Mode of breathing (mouth-open versus mouth-closed) demonstrated itself to have a large impact on the rate of recovery of breath ammonia after a water rinse. Within 30 min, breath ammonia returned to 98 ± 16% that of the baseline with mouth open breathing, while mouth closed breathing allowed breath ammonia to return to 53 ± 14% of baseline. These results contribute to a growing body of literature that will improve reproducibly in ammonia and other VCs.


Comprehensive Characterization of PVDF Nanofibers at Macro- and Nanolevel.
This study is focused on the characterization and investigation of polyvinylidene fluoride (PVDF) nanofibers from the point of view of macro- and nanometer level. The fibers were produced using electrostatic spinning process in air. Two types of fibers were produced since the collector speed (300 rpm and 2000 rpm) differed as the only one processing parameter. Differences in fiber's properties were studied by scanning electron microscopy (SEM) with cross-sections observation utilizing focused ion beam (FIB). The phase composition was determined by Fourier-transform infrared spectroscopy (FTIR) and Raman spectroscopy. The crystallinity was determined by differential scanning calorimetry (DSC), and chemical analysis of fiber's surfaces and bonding states were studied using X-ray photoelectron spectroscopy (XPS). Other methods, such as atomic force microscopy (AFM) and piezoelectric force microscopy (PFM), were employed to describe morphology and piezoelectric response of single fiber, respectively. Moreover, the wetting behavior (hydrophobicity or hydrophilicity) was also studied. It was found that collector speed significantly affects fibers alignment and wettability (directionally ordered fibers produced at 2000 rpm almost super-hydrophobic in comparison with disordered fibers spun at 300 rpm with hydrophilic behavior) as properties at macrolevel. However, it was confirmed that these differences at the macrolevel are closely connected and originate from nanolevel attributes. The study of single individual fibers revealed some protrusions on the fiber's surface, and fibers spun at 300 rpm had a core-shell design, while fibers spun at 2000 rpm were hollow.