Initial Experience of Challenge-Free MRI-Based Oxygen Extraction Fraction Mapping of Ischemic Stroke at Various Stages: Comparison With Perfusion and Diffusion Mapping.
MRI-based oxygen extraction fraction imaging has a great potential benefit in the selection of clinical strategies for ischemic stroke patients. This study aimed to evaluate the performance of a challenge-free oxygen extraction fraction (OEF) mapping in a cohort of acute and subacute ischemic stroke patients. Consecutive ischemic stroke patients (a total of 30 with 5 in the acute stage, 19 in the early subacute stage, and 6 in the late subacute stage) were recruited. All subjects underwent MRI including multi-echo gradient echo (mGRE), diffusion weighted imaging (DWI), and 3D-arterial spin labeling (ASL). OEF maps were generated from mGRE phase + magnitude data, which were processed using quantitative susceptibility mapping (QSM) + quantitative blood oxygen level-dependent (qBOLD) imaging with cluster analysis of time evolution. Cerebral blood flow (CBF) and apparent diffusion coefficient (ADC) maps were reconstructed from 3D-ASL and DWI, respectively. Further, cerebral metabolic rate of oxygen (CMRO2) was calculated as the product of CBF and OEF. OEF, CMRO2, CBF, and ADC values in the ischemic cores (absolute values) and their contrasts to the contralateral regions (relative values) were evaluated. One-way analysis of variance (ANOVA) was used to compare OEF, CMRO2, CBF, and ADC values and their relative values among different stroke stages. The OEF value of infarct core showed a trend of decrease from acute, to early subacute, and to late subacute stages of ischemic stroke. Significant differences among the three stroke stages were only observed in the absolute OEF (F = 6.046, p = 0.005) and relative OEF (F = 5.699, p = 0.009) values of the ischemic core, but not in other measurements (absolute and relative CMRO2, CBF, ADC values, all values of p > 0.05). In conclusion, the challenge-free QSM + qBOLD-generated OEF mapping can be performed on stroke patients. It can provide more information on tissue viability that was not available with CBF and ADC and, thus, may help to better manage ischemic stroke patients.


Ecological effects of salicin at three trophic levels: new problems from old adaptations.
Salicin, a toxic phenol glycoside, is used by larvae of the beetle Chrysomela aenicollis as a substrate for producing defensive secretions. In the east-central Sierra Nevada mountains of California, salicin concentrations ranged from 0.05 percent to over 5 percent of dry weight in leaves of different plants of Salix orestera, the Sierra willow. Beetles produced more secretion and suffered less predation on willows containing more salicin. In addition, leaf damage due to herbivory among 16 willow clones ranged from 0 to 20 percent of leaf area and was linearly related to salicin content. These results illustrate how a plant secondary chemical can become a problem for the plant when herbivores are adapted to use the chemical for their own benefit. The results also show the effect of a plant chemical on three trophic levels-the producer, a herbivore, and the predators of the herbivore.


SERCA2a, phospholamban, sarcolipin, and ryanodine receptors gene expression in children with congenital heart defects.
In animal models of conotruncal heart defects, an abnormal calcium sensitivity of the contractile apparatus and a depressed L-type calcium current have been described. Sarcoplasmic reticulum (SR) Ca(2+) ATPase (SERCA) is a membrane protein that catalyzes the ATP-dependent transport of Ca(2+) from the cytosol to the SR. The activity of SERCA is inhibited by phospholamban (PLN) and sarcolipin (SLN), and all these proteins participate in maintaining the normal intracellular calcium handling. Ryanodine receptors (RyRs) are the major SR calcium-release channels required for excitation-contraction coupling in skeletal and cardiac muscle. Our objective was to evaluate SERCA2a (i.e., the SERCA cardiac isoform), PLN, SLN, and RyR2 (i.e., the RyR isoform enriched in the heart) gene expression in myocardial tissue of patients affected by tetralogy of Fallot (TOF), a conotruncal heart defect. The gene expression of target genes was assessed semiquantitatively by RT-PCR using the calsequestrin (CASQ, a housekeeping gene) RNA as internal standard in the atrial myocardium of 23 pediatric patients undergoing surgical correction of TOF, in 10 age-matched patients with ventricular septal defect (VSD) and in 13 age-matched children with atrial septal defect (ASD). We observed a significantly lower expression of PLN and SLN in TOF patients, while there was no difference between the expression of SERCA2a and RyR2 in TOF and VSD. These data suggest a complex mechanism aimed to enhance the intracellular Ca(2+) reserve in children affected by tetralogy of Fallot.


Cerebellar contributions to verbal working memory.
There is increasing evidence for a cerebellar role in working memory. Clinical research has shown that working memory impairments after cerebellar damage and neuroimaging studies have revealed task-specific activation in the cerebellum during working memory processing. A lateralisation of cerebellar function within working memory has been proposed with the right hemisphere making the greater contribution to verbal processing and the left hemisphere for visuospatial tasks. We used continuous theta burst stimulation (cTBS) to examine whether differences in post-stimulation performance could be observed based on the cerebellar hemisphere stimulated and the type of data presented. We observed that participants were significantly less accurate on a verbal version of a Sternberg task after stimulation to the right cerebellar hemisphere when compared to left hemisphere stimulation. Performance on a visual Sternberg task was unaffected by stimulation of either hemisphere. We discuss our results in the context of prior studies that have used cerebellar stimulation to investigate working memory and highlight the cerebellar role in phonological encoding.


Coherent magnetic plasmon modes in a contacting gold nano-sphere chain on a gold slab.
A coupled magnetic resonator waveguide, composed of a contacting gold nanosphere chain on a gold slab, is proposed and investigated. A broadband coherent magnetic plasmon mode can be excited in this one dimensional nanostructure. By employing the Lagrangian formalism and the Fourier transform method, the dispersion properties of the wave vector and group velocity of the magnetic plasmon mode are investigated. Small group velocity can be obtained from this system which can be applied as subwavelength slow wave waveguides.


Submicron-textured biomaterial surface reduces staphylococcal bacterial adhesion and biofilm formation.
Staphylococci are among the most important pathogens causing bloodstream infections associated with implanted medical devices. Control of bacterial adhesion to material surfaces is important for prevention of biofilm formation and biomaterial-associated infections. In this study, we hypothesized that submicron (staphylococcal bacterial dimension) surface textures may reduce the bacterial adhesion via a decrease in surface area that bacteria can contact, and subsequently inhibit biofilm formation. Poly(urethane urea) films were textured with two different sizes of submicron pillars via a two-stage replication process. Adhesion of two bacterial strains (Staphylococcus epidermidis RP62A and S. aureus Newman) was assessed over a shear stress range of 0-13.2 dyn cm(-2) using a rotating disk system in physiological buffer solutions. Significant decreases in bacterial adhesion were observed on textured surfaces for both strains compared with smooth controls. Biofilm formation was further tested on surfaces incubated in solution for either 2 or 5 days and it was found that biofilm formation was dramatically inhibited on textured surfaces. The results of the approaches used in this work demonstrate that patterned surface texturing of biomaterials provides an effective means to reduce staphylococcal adhesion and biofilm formation on biomaterial surfaces, and thus to prevent biomaterial-associated infections.


Short interpregnancy intervals in the United States.
To investigate the prevalence and correlates of short interpregnancy intervals in the United States. We analyzed pregnancy data from a nationally representative sample of 12,279 women from the 2006-2010 National Survey of Family Growth. We limited our sample to second and higher-order births within 5 years of the interview. Interpregnancy intervals were calculated as the interval between the delivery date of the preceding live birth and the conception date of the index pregnancy, with short interpregnancy intervals defined as intervals less than 18 months. We used simple and multivariate logistic regression analyses to examine associations between short interpregnancy intervals and maternal demographic and childbearing characteristics, including pregnancy intention. Among the 2,253 pregnancies in our sample, one third (35%) were conceived within 18 months of a previous birth. After adjusting for sociodemographic and childbearing characteristics, women were significantly more likely to have a short interpregnancy interval if they were aged 15-19 years or married at the time of conception of the index pregnancy, initiated childbearing after age 30 years, or reported the pregnancy as unintended. Short interpregnancy intervals were more likely to be intended among more advantaged women (married, non-Hispanic white, college-educated, or non-Medicaid delivery). We estimate that preventing unintended pregnancies would reduce the proportion of short interpregnancy intervals from 35% to 23%. Providing counseling about the potential negative consequences of short interpregnancy intervals and improving women's contraceptive use to reduce rates of unintended pregnancy likely would reduce the proportion of short interpregnancy interval pregnancies in the United States. II.


A randomized double-blind, placebo-controlled, cross-over trial (Vestparoxy) of the treatment of vestibular paroxysmia with oxcarbazepine.
Vestibular paroxysmia (VP) is characterized by short, often oligosymptomatic attacks of vertigo which occur spontaneously or are sometimes provoked by turning the head. Despite the description of the disease almost 40 years ago (first termed "disabling positional vertigo"), no controlled treatment trial has been published to date. The Vestparoxy trial was designed as a randomized, placebo-controlled, double-blind cross-over trial to examine the therapeutic effect of oxcarbazepine (OXA) in patients with definite or probable VP. Patients were recruited from August 2005 to December 2011 in the outpatient Dizziness Unit of the Department of Neurology of the Munich University Hospital, and randomized to receive OXA (first week: 300 mg once per day, second week: 300 mg b.i.d., third week: 300 mg t.i.d. until the end of the third month), followed by placebo or vice versa with a 1-month wash-out period in between. The primary endpoint was the number of days with one or more attacks. Secondary endpoints were the number of attacks during the observed days, and the median (for each day) duration of attacks. All these endpoints were assessed using standardized diaries collected at the end of each treatment phase. Forty-three patients were randomized, 18 patients provided usable data (2525 patient days) for at least one treatment phase and were included in the main (intention-to-treat) analysis. The most common reasons for discontinuation documented were adverse events. The risk of experiencing a day with at least one attack was 0.41 under OXA, and 0.62 under placebo treatment, yielding a relative risk of 0.67 (95% CI 0.47-0.95, p = 0.025). The number of attacks during the observed days ratio was 0.53 (95% CI 0.42-0.68, p < 0.001) under OXA compared to placebo. Median attack duration was 4 s (Q25: 2 s, Q75: 120 s) under OXA, and 3 s (Q25: 2 s, Q75: 60 s) under placebo treatment. When days with no attacks, i.e., duration = 0, were included in the analysis, these figures changed to 0 (Q25: 0, Q75: 3 s), and 2 (Q25: 0, Q75: 6 s). No serious adverse events or new safety findings were identified during the trial. The Vestparoxy trial showed a significant reduction of VP attacks under OXA compared to placebo treatment, confirming the known and revealing no new side effects.


Obstructive sleep apnea in association with posterior fossa neurologic disease.
Sleep apnea in association with neurologic disease is uncommon, and its documentation with nocturnal polysomnography (PSG) is rare. Two patients with posterior fossa neurologic disease had obstructive and central sleep apnea on PSG. The first was a 40-year-old woman who experienced a respiratory arrest ten days after neurosurgical decompression of a cervical syrinx associated with syringobulbia. Nocturnal PSG demonstrated obstructive sleep apnea. Tracheostomy was performed and a second nocturnal PSG showed no sleep apnea. The second patient was a 76-year-old woman with daytime hypersomnolence, nocturnal choking spells, progressive spastic paraparesis, and left-sided cerebellar tremor. Neurologic evaluation led to the diagnosis of olivopontocerebellar degeneration. Nocturnal PSG showed multiple episodes of central and obstructive apnea. These cases support the hypothesis that central and obstructive sleep apnea can originate from a central mechanism.


The diversity of protein-protein interaction interfaces within T=3 icosahedral viral capsids.
Some non-enveloped virus capsids assemble from multiple copies of a single type of coat-protein (CP). The comparative energetics of the diverse CP-CP interfaces present in such capsids likely govern virus assembly-disassembly mechanisms. The T = 3 icosahedral capsids comprise 180 CP copies arranged about two-, three-, five- and six-fold axes of (quasi-)rotation symmetry. Structurally diverse CPs can assemble into T = 3 capsids. Specifically, the Leviviridae CPs are structurally distinct from the Bromoviridae, Tombusviridae and Tymoviridae CPs which fold into the classic "jelly-roll" fold. However, capsids from across the four families are known to disassemble into dimers. To understand whether the overall symmetry of the capsid or the structural details of the CP determine virus assembly-disassembly mechanisms, we analyze the different CP-CP interfaces that occur in the four virus families. Previous work studied protein homodimer interfaces using interface size (relative to the monomer) and hydrophobicity. Here, we analyze all CP-CP interfaces using these two parameters and find that the dimerization interface (present between two CPs congruent through a two-fold axis of rotation) has a larger relative size in the Leviviridae than in the other viruses. The relative sizes of the other Leviviridae interfaces and all the jelly-roll interfaces are similar. However, the dimerization interfaces across families have slightly higher hydrophobicity, potentially making them stronger than other interfaces. Finally, although the CP-monomers of the jelly-roll viruses are structurally similar, differences in their dimerization interfaces leads to varied dimer flexibility. Overall, differences in CP-structures may induce different modes of swelling and assembly-disassembly in the T = 3 viruses.


[Chicken pox infection in early pregnancy].
We report on a case of chicken pox in early pregnancy, which occurred despite administration of zoster hyperimmunoglobulin. In this case, a healthy child was born. We discuss the purpose and efficacy of passive immunisation in varicella-zoster infections.


Self-assembly of nickel icosahedrons and truncated octahedral nanocrystals on a SrTiO3 (111) support.
Nickel nanocrystals have received much attention for their ferromagnetic properties. The crystal properties are strongly dependent on their facets and therefore detailed study of their morphology, facets and orientation is critical for magnetic applications. In this work, equilibrium crystal shapes of self-assembled nickel nanocrystals on the (111) termination of strontium titanate (SrTiO3) at room temperature and under ultra-high vacuum (UHV) conditions have been investigated using scanning tunneling microscope. SrTiO3 (111) substrate was sputtered (0.5 keV, 2.5 µA, 10 min) and annealed (900°C, 1 h) under UHV conditions. Three different periodicities were observed: 2.21 ± 0.01 nm corresponding to (4 × 4) reconstruction, 3.31 ± 0.02 nm corresponding to (6 × 6) reconstruction and 2.85 ± 0.05 nm, rotated at 30° with respect to (4 × 4) reconstruction, corresponding to (3√3 × 3√3)R30° reconstruction. Nickel (∼1 ml) was deposited using an e-beam evaporator on the substrate preheated to 320°C and the sample was post-annealed multiple times. Nickel took platonic shapes of supported icosahedron comprising of (111) facets and truncated octahedron comprising of (001) and (111) facets. Based on surface energy ratios of truncated octahedrons at equilibrium, the work of adhesion was calculated to be 3.889 ± 0.167 J/m2.


Manganese carbonyl fluorides: are they viable molecules?
The mononuclear Mn(CO)(5)X and binuclear Mn(2)(CO)(8)(μ-X)(2) manganese carbonyl halides have long been known for the halogens Cl, Br, and I. However, the corresponding manganese carbonyl fluorides (X = F) remain unknown. The structures and thermochemistry of such manganese carbonyl fluorides and their decarbonylation products have now been investigated using density functional theory. In all cases singlet structures were found to have lower energies than the corresponding triplet structures. The expected octahedral structure is predicted for Mn(CO)(5)F. Decarbonylation of Mn(CO)(5)F is predicted to give trigonal bipyramidal Mn(CO)(4)F with equatorial fluorine. Further, decarbonylation gives tetrahedral Mn(CO)(3)F. All of the binuclear Mn(2)(CO)(n)F(2) structures (n = 8, 7, 6) are predicted to have a central Mn(2)F(2) unit with two bridging F atoms, a non-bonding Mn···Mn distance of ~3.1 Å, and exclusively terminal CO groups. The thermochemistry of these manganese carbonyl fluorides indicates that they are viable species. This suggests that the failure to date to synthesize the simple manganese carbonyl fluorides arises from a lack of a suitable synthetic method rather than from the instability of the desired products.


Pneumoproteins as a lung-specific biomarker of alveolar permeability in conventional on-pump coronary artery bypass graft surgery vs mini-extracorporeal circuit: a pilot study.
Despite improvements of the heart-lung machine (HLM), oxidative stress and subsequent damage to the alveolar capillary membrane still occur after conventional on-pump coronary artery bypass graft (CCABG) surgery. In an attempt to further improve the conventional HLM, a mini-extracorporeal circuit (MECC) was introduced. This new concept is based on minimal volume shifts. The extent of alveolar injury that is associated with this new technique is unknown. The lung-specific biomarkers Clara-cell 16 (CC16) and KL-6 are applied in this study to quantify alveolar dysfunction in both techniques. In a prospective observational setting, the concentrations of CC16 and KL-6 were measured during and after 10 consecutive CCABG operations and 10 consecutive coronary artery bypass graft (CABG) operations using MECC (MCABGs). These pneumoproteins were measured after the induction of anesthesia, before clamping of the ascending aorta, after unclamping of the aorta, on arrival to the ICU, and on the following days until discharge. Quantification of the differences of KL-6 and CC16 leakage through the alveolar membranes between the two techniques was realized by calculation of the Student t test. Perioperative and postoperative shunt fractions and clinical observations were monitored simultaneously. The potential value of pneumoproteins as biomarkers for quantification of alveolar permeability during CABG surgery was tested. Significantly reduced concentrations of CC16 were found early after MCABG as compared to CCABG surgery (p = 0.033). KL-6 showed no consistent pattern during both treatment modalities. Early after CCABG surgery, shunt fractions tended to show reduced oxygen transport over the alveolar membrane as compared to MCABG surgery. CC16 appears to be a useful biomarker for alveolar permeability during CABG surgery. Injury of the alveolar capillary membrane appears significantly reduced during MCABG surgery. Consistently early postoperative alveolar shunt fractions showed an increased value in CCABG compared to MCABG surgery in the early postoperative phase. Further randomized studies need to confirm the value of CC16 as marker in monitoring alveolar capillary damage during coronary bypass grafting.


Escaping the box: preparing allied health practitioners for management positions.
Career ladders designed for health care practitioners are not always long enough to reach beyond the top rungs of clinical practice. Limited upward mobility can result in dissatisfaction and potential loss of experienced staff members. The same skills that allow practitioners to function successfully in patient care can be tapped into to prepare clinicians to move upward in administrative positions. This has already occurred successfully in the nursing profession. Managers should take advantage of the yearly performance evaluation to identify those health professionals who are interested in advancement. Using a strengths, weakness, opportunities, and threats analysis, the manager and clinician can work together to develop a developmental plan that will build on existing strengths while developing the less prepared areas. Managerial skill enhancement should be combined with mentoring from persons already filling administrative roles to enable the clinician to develop a network of contacts. A planned approach should enable interested clinicians to successfully transition from hands-on practice into managerial positions.


[Coronary angioplasty today].
The rise of coronary angioplasty has been rapid in the last ten years. It is a method of myocardial revascularisation without thoracotomy or cardiopulmonary bypass and is curative in the absence of restenosis. Recent advances concern the development of the material and the pharmacological support which reduce the risk of coronary thrombosis during the procedure. The primary success rate now approaches 90%, that of emergency bypass surgery less than 5%. The use of autoperfusion catheters and the use of intra-aortic balloon pumping improve the safety of the procedure but surgical stand-by remains a necessity in cases of dilatation of proximal main coronary stenosis. Stable and unstable angina are the main indications of a technique particularly appropriate for elderly patients and the treatment of residual post-thrombolytic stenoses, post-radic stenosis or stenosis of a single functional coronary artery. Restenosis is observed in 30 to 40% of cases during the first six months after the dilatation. Its occurrence is not influenced by any of the drugs tested but it could be reduced by the insertion of an endocoronary stent. Abrasive endocoronary techniques and laser angioplasty are alternative methods of treating coronary stenoses inaccessible to conventional balloon angioplasty but they do not reduce the risk of restenosis.


New scientific approaches to cancer treatment: can medicinal mushrooms defeat the curse of the century?
Modern science has spread so broadly and deeply within the nature of organisms that researchers have already established the molecular mechanisms of each vital process of human existence. Unfortunately, our age, heredity, lifestyle, race, and ethnicity are directly connected to the development and expression of many negative and even fatal processes, such as carcinogenesis. Many investigators have shown that the roots of carcinogenesis are hidden in the signaling pathways of cancer cells. Therefore, the pharmacological modulation of intracellular signal transduction pathways and the activation of transcription factors regulating gene expression represent a new and attractive therapeutic approach to cancer. It is already well established that medicinal mushrooms represent a potential source for biologically active compounds with immunomodulating, free radical scavenging, anti-inflammatory, antibacterial, antifungal, antiviral, hepatoprotective, antidiabetic, and anticancer activities. Moreover, low-molecular weight mushroom substances have been studied, and reported to interact with particular intracellular signaling pathways related to processes such as inflammation, cell differentiation and survival, apoptosis, angiogenesis, tumor progression and metastasis, etc. The present work compiles the available data on mushroom substances, in the form of extracts or chemically defined compounds, known to interfere with the above-mentioned essential processes related to the carcinogenesis and, thus, representing a new scientific approach to cancer treatment and prevention.


Comparative HPLC and ELISA studies for CDT isoform characterization in subjects with alcohol related problems. Prospective application in workplace risk-prevention policy.
Serum carbohydrate-deficient transferrin (CDT) is the most specific marker of chronic alcohol abuse so far. The performance of commercial HPLC over the ELISA method for measurement of CDT was evaluated on a series of 105 serum samples obtained from subjects referred to the Toxicology Laboratory of Salvatore Maugeri Hospital for alcohol-related problems. Compared to ELISA, HPLC analysis was more valuable for determining alcohol-related patterns of CDT isoforms and quantifying serum levels of disialotransferrin that better reflect chronic heavy drinking. Other significant advantages of the HPLC method included reproducible separation and easier detection of glycoform types and genetic transferrin variants that are known to cause falsely high or low results in sera examined by immunoassay. Current scientific evidence indicates that disialotransferrin is the target analyte for CDT determination and HPLC the current CDT analysis reference method. Systematic studies for early assessment of excessive alcohol intake or abuse of alcoholic substances in workers are recommended by the Italian legislation in accordance with the European Alcohol Action Plan (EAAP) launched by the WHO Regional Committee for Europe. These studies are advisable given their potential role in preventing negative effects of alcohol abuse in workplace. A research strategy combining CDT and other laboratory markers with questionnaire and physician interview is recommended for examining subjects with alcohol related problems and the diagnosis of alcoholism. This approach can be applied for alcohol abuse in workplace surveillance.


Characteristic differences between vestibular migraine and migraine only patients.
Vestibular migraine (VM) is one of the most common causes of vertigo in clinical practice but it is not always easy to make the correct diagnosis. Our aims were to find out how VM patients differ from migraine only (MO) patients, to evaluate co-morbid depression in these two groups and to determine if their disease has an effect on their quality of life. We studied 50 definite VM and 35 MO patients. Each patient was asked about: age of onset, duration of headaches, presence of aura, headache characteristics, triggering factors, associated features, motion sickness history and family history of migraine. VM patients were also asked about their vertigo attacks and accompanying symptoms. Each patient also completed the following questionnaires: (1) Migraine Disability Assessment Scale (MIDAS); (2) headache severity with VAS (Visual Analog Scale); (3) Allodynia Symptom Checklist (ASC-12); (4) Beck Depression Inventory (BDI); (5) World Health Organization Quality of Life Questionnaire Short Form-12 (WHOQL-SF12); (6) Activities Specific Balance Confidence Scale (ABC). VM patients also completed the Dizziness Handicap Inventory (DHI). We found that VM patients were more likely than MO patients to be female, post-menopausal, depressed, motion sick, complaining of imbalance and of food-triggered headaches. In contrast, MO patients were more likely than VM patients to have severe headaches and that these can be triggered by certain odors and by noise. Our findings showed differences between VM and MO patients and attention to these differences could help clinicians diagnose, characterize and manage their VM patients.


Dialect-neutral indices of narrative cohesion and evaluation.
This study compared the development of essential elements of narrative skill in children from African American English (AAE)- and general American English (GAE)-speaking communities using an innovative elicitation and evaluation protocol consisting of four key indices of narrative language: (a) reference contrasting, (b) temporal expressions, (c) mental state descriptions, and (d) understanding of behavior based on false belief. Participants were 291 AAE speakers and 238 GAE speakers, 4 to 9 years of age. Approximately one-third of both dialect groups were identified as having language impairments. Children generated 2 stories based on short picture sequences. Their stories were coded for the 4 key indices of narrative language. Analyses of variance were performed with subsets of the measures and a composite index with all measures combined as outcomes; and with age, dialect group, and clinical status as predictors. Age and clinical status had statistically significant effects on the subset measures and the composite score. Variation between AAE and GAE dialect was not a significant factor. By focusing on dialect-neutral elements of narratives--creating links across sentences and providing mental state interpretations--this study adds to our knowledge of development and impairment in narrative production among both AAE- and GAE-background children.


Kinetics and mechanism of oxidation of tryptophan by ferrate(VI).
Kinetics of the oxidation of tryptophan (Trp) and kynurenine (Kyn), precursors of nitrogenous disinfection byproducts (N-DBP), by ferrate(VI) (Fe(VI)O4(2-), Fe(VI)) were investigated over the acidic to basic pH range. The second-order rate constants decreased with increase in pH, which could be described by the speciation of Fe(VI) and Trp (or Kyn). The trend of pH dependence of rates for Trp (i.e., aromatic α-amino acid) differs from that for glycine (i.e., aliphatic α-amino acid). A nonlinear relationship between transformation of Trp and the added amount of Fe(VI) was found. This suggests that the formed intermediate oxidized products (OPs), identified by LC-PDA and LC-MS techniques, could possibly compete with Trp to react with Fe(VI). N-Formylkynurenine (NFK) at pH 7.0 and 4-hydroxyquinoline (4-OH Q) and kynurenic acid (Kyn-A) at pH 9.0 were the major OPs. Tryptophan radical formation during the reaction was confirmed by the rapid-freeze quench EPR experiments. The oxygen atom transfer from Fe(VI) to NFK was demonstrated by reacting Fe(18)O4(2-) ion with Trp. A proposed mechanism explains the identified OPs at both neutral and alkaline pH. Kinetics and OPs by Fe(VI) were compared with other oxidants (chlorine, ClO2(•), O3, and (•)OH).


Assessing Genetic Diversity to Breed Competitive Biofortified Wheat With Enhanced Grain Zn and Fe Concentrations.
Breeding wheat with enhanced levels of grain zinc (Zn) and iron (Fe) is a cost-effective, sustainable solution to malnutrition problems. Modern wheat varieties have limited variation in grain Zn and Fe, but large-scale screening has identified high levels of Zn and Fe in wild relatives and progenitors of cultivated wheat. The most promising sources of high Zn and Fe are einkorn (Triticum monococcum), wild emmer (T. dicoccoides), diploid progenitors of hexaploid wheat (such as Aegilops tauschii), T. spelta, T. polonicum, and landraces of T. aestivum. This study evaluate the effects of translocations from rye and different Aegilops species in a "Pavon-76" wheat genetic background and utilized in the wheat biofortification breeding program at CIMMYT that uses diverse genetic resources, including landraces, recreated synthetic hexaploids, T. spelta and pre-breeding lines. Four translocations were identified that resulted significantly higher Zn content in "Pavon 76" genetic background than the check varieties, and they had increased levels of grain Fe as well-compared to "Pavon 76." These lines were also included in the breeding program aimed to develop advanced high Zn breeding lines. Advanced lines derived from diverse crosses were screened under Zn-enriched soil conditions in Mexico during the 2017 and 2018 seasons. The Zn content of the grain was ranging from 35 to 69 mg/kg during 2017 and 38 to 72 mg/kg during 2018. Meanwhile grain Fe ranged from 30 to 43 mg/kg during 2017 and 32 to 52 mg/kg during 2018. A highly significant positive correlation was found between Zn and Fe (r = 0.54; P < 0.001) content of the breeding lines, therefore it was possible to breed for both properties in parallel. Yield testing of the advanced lines showed that 15% (2017) and 24% (2018) of the lines achieved 95-110% yield potential of the commercial checks and also had 12 mg/kg advantage in the Zn content suggesting that greater genetic gains and farmer-preferred wheat varieties were developed and deployed. A decade of research and breeding efforts led to the selection of "best-bet" breeding lines and the release of eight biofortified wheat varieties in target regions of South Asia and in Mexico.


Digestibility and metabolizable energy of raw soybeans manufactured with different processing treatments and fed to adult dogs and puppies.
The objective of this study was to evaluate the apparent total tract digestibility (ATTD), ME, and fecal characteristics of adult dogs and puppies fed raw soybeans (RSB) and their by-products. Six treatments were evaluated: 1 reference diet (REF), based on a maize-poultry by-product meal, and 5 extruded diets containing 70% of the ingredients of the REF diet and 30% of a soybean processed product [defatted soybean meal (DSM), micronized soybeans (MSB), soybean meal (SBM), RSB, or toasted soybeans (TSB)]. Six adult dogs (5.8 yr old) and 6 puppies (5.1 mo old) were used in a study with a double Latin square design (6 × 6). Urease was reduced in all diets after extrusion, but trypsin inhibitor was reduced only in the diets containing SBM, DSM, and RSB. The ATTD of CP in DSM, SBM, MSB, TSB, and RSB were 85.1%, 85.2%, 88.4%, 84.7%, and 78.9%, respectively, for adult dogs. Soybean meal and DSM had the lowest ATTD of acid-hydrolyzed fat (AHF; 84.3% for both ingredients in adult dogs). The ATTD of DM and AHF in DSM and AHF in all soybean products were greater in puppies than adult dogs (P < 0.05). The ME content was greatest in MSB (21.39 MJ/kg) and least in DSM (15.23 MJ/kg). The feces of dogs fed soybean products were softer and had a lower pH (average of 5.91 vs. 6.05 for adult dogs fed soybean products and REF diets, respectively) and ammonia content (average of 3.82 vs. 4.32 g/kg for adult dogs fed soybean products and REF diets, respectively), except those fed RSB, which had similar fecal pH and ammonia values, compared with those fed the REF diet. Soybean products are good protein sources for both adult and growing dogs, provided they are heat treated before diet extrusion.


The differential control of c-jun expression in regenerating sensory neurons and their associated glial cells.
Damage to the axons of adult sensory neurons results in massively increased expression of the protooncogene c-jun both in neurons and in the associated Schwann cells. The role of growth factors and axon contact in mediating this expression was investigated in dissociated cultures of adult sensory neurons and glial cells that expressed c-jun within 24 hr of plating. Trk, trkB, and trkC growth factor receptor genes were expressed in discrete subpopulations of sensory neurons but addition of NGF or brain-derived neurotrophic factor (BDNF) did not inhibit the expression of c-jun. Similarly, axon contact was not sufficient to decrease c-jun expression in glial cells. However, c-jun expression could be downregulated in glial cells, but not neurons, by treatment with cAMP or forskolin and increased by raising intracellular calcium levels. The results suggest that c-jun levels are differentially regulated in neurons and glial cells but that NGF or BDNF do not regulate c-jun expression in damaged neurons.


Chronic hepatitis C and superimposed nonalcoholic fatty liver disease.
Hepatitis C and nonalcoholic fatty liver disease (NAFL) are the two most common forms of liver disease in the United States. Recently, obesity and its associated risk factors have been suggested to enhance HCV-related fibrosis. The aim of this study was to assess the impact of hepatic steatosis, steatohepatitis, and its associated risk factors on HCV-related fibrosis. Patients with untreated, biopsy-proven, chronic hepatitis C (6/97-3/99) were included. Clinical and demographic data at the time of liver biopsy were obtained from chart review and verified by telephone survey. One hepatopathologist reviewed all pathologic specimens, using the modified histological activity index score and the Ishak staging for fibrosis and a NAFL pathologic protocol. One hundred and seventy patients with hepatitis C were included [age: 48.7+/-9.33 (years), body mass index (BMI): 28.1+/-5.7 (kg/m2) and type 2 diabetes mellitus (DM): 14%]. Of these, 77 (45.3%) had no or mild fibrosis and 93 (54.7%) had advanced fibrosis. Hepatic steatosis was seen in 90 (52.9%) patients. The grade of steatosis was associated with markers of obesity only. Age (p=0.002), type 2 DM (p=0.04), and superimposed steatohepatitis (p=0.047) were independently associated with advanced fibrosis. Superimposed nonalcoholic steatohepatitis (NASH) was seen in 17 (10%) patients. Patients with superimposed NASH were mostly obese (76.5%), males (62%) with 16% having type 2 diabetes and a BMI 33.8+/-7.12. In patients with chronic hepatitis C, type 2 DM and superimposed steatohepatitis are independently associated with advanced fibrosis.


Generalized Indeterminate Cell Histiocytosis Presenting as Eroded Papules and Crusts.
Indeterminate cell histiocytosis (ICH) is a very rare histiocytic disease. We reported a case of generalized ICH presenting as eroded papules and crusts, accompanied with intermittent fever, pulmonary disorder, lymphadenectasis in the mediastinum, and the retroperitoneum. The histopathology indicated a neoplasm that was composed of small nests of large cells with ample pale cytoplasm. Immunohistochemistry results and the absence of Birbeck granules led us to the diagnosis of ICH. Four months later, the patient died of anemia, ascites, and swelling of the limbs.


First demonstration of MC-EDFA-repeatered SDM transmission of 40 x 128-Gbit/s PDM-QPSK signals per core over 6,160-km 7-core MCF.
We demonstrate the first 7-core multicore erbium-doped fiber amplified (MC-EDFA) transmission of 40 x 128-Gbit/s PDM-QPSK signals over 6,160-km 7-core multicore fiber (MCF). The crosstalk (XT) from all of the other 6 cores of a MC-EDFA and a 55-km length MCF are about -46.5 dB and -45.6 dB at center core, respectively. The core-to-core rotation approach at every amplified span is used to average the XT of all cores. The averaged optical signal-to-noise ratio (OSNR) after 6,160-km transmission is 15.6 dB with 0.1 nm resolution bandwidth. The Q-factor of all 40 channels surpasses the threshold of the forward-error-correction of 6.4 dB with 1 dB margin after 6,160 km. The total net capacity is 28.8 Tbit/s per fiber and achieved capacity-distance product is 177 Pbit/s.km per fiber. We confirmed the feasibility of MC-EDFA repeatered systems for trans-oceanic transmission.


Pulsed electromagnetic fields simultaneously induce osteogenesis and upregulate transcription of bone morphogenetic proteins 2 and 4 in rat osteoblasts in vitro.
Pulsed electromagnetic fields (PEMF) are successfully employed in the treatment of a variety of orthopaedic conditions, particularly delayed and nonunion fractures. In this study, we examined PEMF effects on in vitro osteogenesis by bone nodule formation and on mRNA expression of bone morphogenetic proteins 2 and 4 by reverse-transcriptase polymerase chain reaction (RT-PCR) in cultured rat calvarial osteoblasts. PEMF exposure induced a significant increase in both the number (39% over unexposed controls) and size (70% larger compared to unexposed controls) of bone-like nodules formed. PEMF also induced an increase in the levels of BMP-2 and BMP-4 mRNA in comparison to controls. This effect was directly related to the duration of PEMF exposure. This study shows that clinically applied PEMF have a reproducible osteogenic effect in vitro and simultaneously induce BMP-2 and -4 mRNA transcription. This supports the concept that the two effects are related.


Quantification and Imaging of Antigens on Cell Surface with Lipid-Encapsulated Fluorescent Nanodiamonds.
Quantifying the density and locating the position of antigens on cell surface has been a challenge in molecular biology research. The challenge lies in the need for a chemically and photophysically stable fluorophore to achieve the required sensitivity and accuracy. Here, we present a method suitable for the purpose by using lipid-encapsulated fluorescent nanodiamonds (FNDs) of 35 nm in diameter as biolabels. The encapsulation of FNDs in biotinylated phospholipids not only facilitates good dispersion of the particles in biological buffers, but also endows them with high specific targeting ability. We demonstrated a viable application of the technique for biotin-mediated immunostaining of antigens on fixed human cells, identifying their positions by two-color confocal fluorescence imaging, and determining their densities by magnetically modulated fluorescence detection. A binding capacity of 6 ± 1 × 104 antigens/cell was measured specifically for CD44 on HeLa cell surface. The result agreed well with the assay of R-phycoerythrin-conjugated antibodies by flow cytometry, supporting the reliability of this new nanoparticle-based method.


Prevention of oral biofilm formation and degradation of biofilm by recombinant α-1,3-glucanases from Streptomyces thermodiastaticus HF3-3.
The genes encoding α-1,3-glucanases (Agls; AglST1 and AglST2) from Streptomyces thermodiastaticus HF3-3 were cloned and were then expressed in Escherichia coli Rosetta-gami B (DE3). We purified the resultant histidine (His)-tagged α-1,3-glucanases (recombinant enzymes, rAglST1 and rAglST2). Both the recombinant enzymes were similar to the wild-type enzymes. We examined the effects of rAglST1 and rAglST2 on the formation and degradation of biofilms on glass plates with Streptococcus mutans NRBC 13955 by evaluating the biofilm content (%), release of reducing sugar (mM), release of S. mutans (log CFU/mL), and the biofilm structure using laser scanning microscopy (LSM). The results showed that after incubation for 16 h, rAglST1 and rAglST2 reduced the formation of biofilm to 52% and 49% of the control, respectively. The result may reflect the fact that the concentration of the reducing sugar and the number of S. mutans cells in the rAglATs-added medium were higher than in the control medium. After an 8-h treatment with rAglST1 and rAglST2, biofilms decreased to less than 60% of the control. The number of S. mutans cells in the reaction mixture gradually increased during the incubation period. The enzymes can degrade the biofilms that were pre-formed on the glass plate by more than 50% after a 30-min incubation in the presence of toothpaste ingredients (1% w/v of sodium fluoride, benzethonium chloride, and sodium dodecyl sulfate) at 50°C. Our study showed that rAglST1 and rAglST2 have advantageous properties for dental care applications.


Knee biomechanics early after knee replacement surgery predict abnormal gait patterns 12 months postoperatively.
An abnormal flexor moment pattern is often evident following total knee replacement (TKR) surgery. We investigated whether such a pattern at 12 months post-surgery could be predicted using biomechanical gait measures assessed before surgery and at 4 months post-surgery. Thirty two TKR patients were evaluated and classified as normal (biphasic pattern) or abnormal (flexor moment pattern) at each time point. Biomechanical parameters collected before surgery and at 4 months post-surgery were then explored for their ability to predict gait patterns at 12 months post-surgery. The gait pattern at 4 months was significantly associated with the 12 month post-surgery gait pattern, with over half of those with a flexor moment pattern at 4 months retaining this pattern at 12 months. Discriminant function analysis indicated that peak knee flexion during early stance, peak knee extension, and peak knee extension moment at 4 months post-surgery were independent predictors of the gait pattern at 12 months. Thus, an abnormal knee flexor moment pattern at 12 months post-surgery can be predicted by biomechanical analysis 4 months after surgery. Therefore, interventions aimed at improving active extension may need to be implemented early after surgery to restore a normal gait pattern.


Polatuzumab vedotin, an anti-CD79b antibody-drug conjugate for the treatment of relapsed/refractory diffuse large B-cell lymphoma.
Refractory/relapsed diffuse large B-cell lymphoma remains a major unmet medical need with poor outcome, especially for patients considered ineligible for stem cell transplant. Polatuzumab vedotin (PV) is a first-in-class anti-CD79b antibody-drug conjugate that contains the microtubule inhibitor monomethyl auristatin E. The development of PV is currently very active. This drug was US FDA approved in 2019 in combination with bendamustine and rituximab for the treatment of refractory/relapsed diffuse large B-cell lymphoma in third line and more, after demonstrating relevant efficacy and acceptable safety in a pivotal randomized Phase II trial. This review summarizes the features of this new drug with the primary focus on the clinical work supporting efficacy, relevance and tolerability of PV.


Electrochemical mechanism of ion-ionophore recognition at plasticized polymer membrane/water interfaces.
Here, we report on the first electrochemical study that reveals the kinetics and molecular level mechanism of heterogeneous ion-ionophore recognition at plasticized polymer membrane/water interfaces. The new kinetic data provide greater understanding of this important ion-transfer (IT) process, which determines various dynamic characteristics of the current technologies that enable highly selective ion sensing and separation. The theoretical assessment of the reliable voltammetric data confirms that the dynamics of the ionophore-facilitated IT follows the one-step electrochemical (E) mechanism controlled by ion-ionophore complexation at the very interface in contrast to the thermodynamically equivalent two-step electrochemical-chemical (EC) mechanism based on the simple transfer of an aqueous ion followed by its complexation in the bulk membrane. Specifically, cyclic voltammograms of Ag(+), K(+), Ca(2+), Ba(2+), and Pb(2+) transfers facilitated by highly selective ionophores are measured and analyzed numerically using the E mechanism to obtain standard IT rate constants in the range of 10(-2) to 10(-3) cm/s at both plasticized poly(vinyl chloride) membrane/water and 1,2-dichloroethane/water interfaces. We demonstrate that these strongly facilitated IT processes are too fast to be ascribed to the EC mechanism. Moreover, the little effect of the viscosity of nonaqueous media on the IT kinetics excludes the EC mechanism, where the kinetics of simple IT is viscosity-dependent. Finally, we employ molecular level models for the E mechanism to propose three-dimensional ion-ionophore complexation at the two-dimensional interface as the unique kinetic requirement for the thermodynamically facilitated IT.


Tackling Covid-19 using disordered-to-order transition of residues in the spike protein upon angiotensin-converting enzyme 2 binding.
The 2019-novel coronavirus also known as severe acute respiratory syndrome-coronavirus 2 (SARS-CoV-2) is a common threat to animals and humans, and is responsible for the human SARS pandemic in 2019 to 2021. The infection of SARS-CoV-2 in humans involves a viral surface glycoprotein named as spike proteins, which bind to the human angiotensin-converting enzyme 2 (ACE2) proteins. Particularly, the receptor binding domains (RBDs) mediate the interaction and contain several disordered regions, which help in the binding. Investigations on the influence of disordered residues/regions in stability and binding of spike protein with ACE2 help to understand the disease pathogenesis, which has not yet been studied. In this study, we have used molecular-dynamics simulations to characterize the structural changes in disordered regions of the spike protein that result from ACE2 binding. We observed that the disordered regions undergo disorder-to-order transition (DOT) upon binding with ACE2, and the DOT residues are located at functionally important regions of RBD. Although the RBD is having rigid structure, DOT residues make conformational rearrangements for the spike protein to attach with ACE2. The binding is strengthened via hydrophilic and aromatic amino acids mainly present in the DOTs. The positively correlated motions of the DOT residues with its nearby residues also explain the binding profile of RBD with ACE2, and the residues are observed to be contributing more favorable binding energies for the spike-ACE2 complex formation. This study emphasizes that intrinsically disordered residues in the RBD of spike protein may provide insights into its etiology and be useful for drug and vaccine discovery.


Lipopeptide PAM3CYS4 Synergizes N-Formyl-Met-Leu-Phe (fMLP)-Induced Calcium Transients in Mouse Neutrophils.
N-Formyl-Met-Leu-Phe (fMLP), a mimic of N-formyl oligopeptides that are released from bacteria, is a potent leukocyte chemotactic factor. It induces intracellular calcium ([Ca]i) transient that is important for various neutrophil biological functions, e.g., adhesion, ROS, and cytokine productions. Toll-like receptors (TLRs), an essential part of host innate immunity, regulate neutrophil activities, but their role in [Ca]i signaling is less clear. In the present study, we examined the effect of several TLR ligands, including Pam3Cys4 (TLR1/2), lipopolysaccharide (LPS, TLR4), and lipoteichoic acid (LTA, TLR2/6), on calcium signaling and on the fMLP-induced [Ca]i transients in mouse neutrophils loaded with Fura-2/AM. We found that unlike fMLP, the three TLR ligands tested did not elicit any detectable Ca flux. However, Pam3Cys4, but not LPS or LTA, markedly synergized the fMLP-induced [Ca]i transients, and had no effect on the host component keratinocyte-derived cytokine (KC)- or C5a-induced calcium flux. The effect of Pam3Cys4 on the fMLP-induced [Ca]i transients is by enhancing extracellular Ca influx, not intracellular Ca release. Surprisingly, deletion of TLR2 or MyD88 in neutrophils had no impact on the Pam3Cys4's effect, suggesting a TLR2-MyD88-independent mechanism. Finally, using the pan PKC activator and inhibitor, we demonstrated that PKC negatively regulated fMLP-induced [Ca]i transients and that inhibition of PKC did not prohibit Pam3Cys4's synergistic effect on the fMLP-induced calcium influx. In conclusion, the present study identified a novel synergistic effect of Pam3Cys4 on fMLP-induced [Ca]i transients, a process important for many neutrophil biological functions.


Fingolimod-associated PML with mild IRIS in MS: A clinicopathologic study.
To clarify the clinical, neuropathologic, and virologic characteristics of progressive multifocal leukoencephalopathy (PML) and its immune reconstitution inflammatory syndrome (IRIS) in a patient with fingolimod-treated MS. A case study. A 34-year-old patient with MS using fingolimod for 4 years had a gradual progression of right hemiparesis and aphasia with a new subcortical white matter lesion in the precentral gyrus by initial MRI. Blood tests were normal, except for lymphopenia (160 cells/μL). One month after the cessation of fingolimod, brain MRI depicted a diffusely exacerbated hyperintensity on fluid-attenuated inversion recovery and diffusion-weighed imaging in the white matter with punctate gadolinium enhancement, suggesting PML-IRIS. A very low level of JC virus (JCV)-DNA (15 copies/mL) was detected in the CSF as judged by quantitative PCR. Brain tissues were biopsied from the left frontal lesion, which showed some small demyelinated foci with predominant loss of myelin-associated glycoprotein with infiltrations of lymphocytes and macrophages, but clear viral inclusion was not observed with hematoxylin-eosin staining. JCV-DNA was uniquely detectable in an active inflammatory demyelinating lesion by in situ hybridization, possibly suggesting an early phase of PML. DNA extracted from the brain sample was positive for JCV-DNA (151 copies/cell). It took 3 months to normalize the blood lymphocyte count. The patient was treated with 1 g of IV methylprednisolone for 3 days and a weekly oral dose (375 mg) of mefloquine, and her symptoms gradually improved. Low CSF JCV-DNA and unfound viral inclusions initially made her diagnosis difficult. The clinical course of fingolimod-associated PML may be associated with mild immune reconstitution.


Characterization of lead-phytochelatin complexes by nano-electrospray ionization mass spectrometry.
The role of phytochelatins (PC(n), metal-binding oligopeptides with the general structure (γGlu-Cys)(n)-Gly (n = 2-11) in metal detoxification is assumed to be based on immobilization of metals, which prevents binding of metals to important biomolecules. Although induction of phytochelatin synthesis has often been observed in algae upon exposure to metals, direct evidence for binding of the inducing metal to phytochelatins is scarce. In this study, a nano-electrospray ionization mass spectrometry (nano-ESI-MS) method is developed for identification and characterization of Pb(II)-PC(n) and Zn(II)-PC(n) complexes. Complexes of Pb(II) with standard PC(n) (n = 2-4; 0.25 mM Pb(II) and 0.5 mM PC(n)) were examined by nano-ESI-MS with respect to their stoichiometry. Pb-PC(n) mass spectra indicated the presence of the [M + H](+) peak of PC(n) and complexes with various stoichiometries. Analysis of Pb-PC(2) allowed the identification of four different complexes observed at m/z 746.10, 952.06, 1285.24, and 1491.20, corresponding to [Pb-PC(2)](+), [Pb(2)-PC(2)](+), [Pb-(PC(2))(2)](+), and [Pb(2)-(PC(2))(2)](+). Their m/z indicated coordination of Pb(II) by PC(2) through the thiol groups of PC cysteine and possibly carboxylic groups. For each of the standard PC(3) and PC(4), two different complexes were observed, corresponding to Pb-PC(3), Pb(2)-PC(3), Pb-PC(4), and Pb(2)-PC(4). The measured isotopic patterns were for all complexes identical to the theoretical isotopic patterns. Addition of Zn(II) (0.125-5 mM) to previously formed Pb-PC(2) complexes showed the appearance of the [Zn-PC(2)](+) complexes at m/z 602.05 and the decrease of the [Pb-PC(2)](+) peak. These findings corroborate the postulated Pb-PC complexes from a previous study using size exclusion chromatography of PC extracted from algae, as well as the concurrent formation of Pb-, Zn-, and Cu-PC complexes in algae.


The endocannabinoid hydrolase FAAH is an allosteric enzyme.
Fatty acid amide hydrolase (FAAH) is a membrane-bound homodimeric enzyme that in vivo controls content and biological activity of N-arachidonoylethanolamine (AEA) and other relevant bioactive lipids termed endocannabinoids. Parallel orientation of FAAH monomers likely allows both subunits to simultaneously recruit and cleave substrates. Here, we show full inhibition of human and rat FAAH by means of enzyme inhibitors used at a homodimer:inhibitor stoichiometric ratio of 1:1, implying that occupation of only one of the two active sites of FAAH is enough to fully block catalysis. Single W445Y substitution in rat FAAH displayed the same activity as the wild-type, but failed to show full inhibition at the homodimer:inhibitor 1:1 ratio. Instead, F432A mutant exhibited reduced specific activity but was fully inhibited at the homodimer:inhibitor 1:1 ratio. Kinetic analysis of AEA hydrolysis by rat FAAH and its F432A mutant demonstrated a Hill coefficient of ~1.6, that instead was ~1.0 in the W445Y mutant. Of note, also human FAAH catalysed an allosteric hydrolysis of AEA, showing a Hill coefficient of ~1.9. Taken together, this study demonstrates an unprecedented allosterism of FAAH, and represents a case of communication between two enzyme subunits seemingly controlled by a single amino acid (W445) at the dimer interface. In the light of extensive attempts and subsequent failures over the last decade to develop effective drugs for human therapy, these findings pave the way to the rationale design of new molecules that, by acting as positive or negative heterotropic effectors of FAAH, may control more efficiently its activity.


Antibiotic resistance in Acinetobacter spp. isolated from sewers receiving waste effluent from a hospital and a pharmaceutical plant.
The possible increase of antibiotic-resistant bacteria in sewage associated with the discharge of wastewater from a hospital and a pharmaceutical plant was investigated by using Acinetobacter species as environmental bacterial indicators. The level of susceptibility to six antimicrobial agents was determined in 385 Acinetobacter strains isolated from samples collected upstream and downstream from the discharge points of the hospital and the pharmaceutical plant. Results indicated that while the hospital waste effluent affected only the prevalence of oxytetracycline resistance, the discharge of wastewater from the pharmaceutical plant was associated with an increase in the prevalence of both single- and multiple-antibiotic resistance among Acinetobacter species in the sewers.


IGF1 does not overcome sexual dimorphism of body and muscle size in Mstn-/- mice.
Insulin-like growth factor-1 (IGF1) is crucial for regulating post-natal growth and, along with myostatin (MSTN), regulates muscle size. Here, we sought to clarify the roles of these two genes in regulating sexually dimorphic growth of body and muscle mass. In the first study, we established that Igf1 mRNA was increased to a greater extent and Igf1 receptor mRNA increased earlier in male, than in female, gastrocnemius muscles during the rapid phase of growth (from 2 to 6 weeks) were unchanged, thereafter, to 32 weeks of age in WT mice (P < 0.001). In the second study, we sought to determine if supplemental IGF1 could overcome the sexual dimorphism of muscle and body mass, when myostatin is absent. We crossed myostatin null (Mstn-/-) mice with mice over-expressing Igf1 in skeletal muscle (Igf1+) to generate six genotypes; control (Mstn+/+), Mstn+/-, Mstn-/-, Mstn+/+:Igf1+, Mstn+/-:Igf1+ and Mstn-/-:Igf1+ (n = 8 per genotype and sex). In both sexes, body mass at 12 weeks was increased by at least 1.6-fold and muscle mass by at least 3-fold in Mstn-/-:Igf1+ compared with Mstn+/+ mice (P < 0.001). The abundance of AKT was increased in muscles of mice transgenic for Mstn, while phosphorylation of AKTS473 was increased in both male and female mice transgenic for Igf1+. The ratio of phosphorylated to total AKT was 1.9-fold greater in male mice (P < 0.001). Thus, despite increased growth of skeletal muscle and body size when myostatin was absent and IGF1 was in excess, sexual dimorphism persisted, an effect consistent with greater IGF1-induced activation of AKT in skeletal muscles of males.


Training and generalized production of wh- and NP-movement structures in agrammatic aphasia.
The present research examines production of "complex" sentences, which involve movement of noun phrases (NPs), in 2 agrammatic aphasic subjects. According to linguistic theory (Chomsky, 1991, 1993), such sentences are derived using one of two movement operations, either wh- or NP-movement, subsumed under the general rule "move alpha." In this experiment recovery of both wh- and NP-movement derived sentences was investigated using a treatment research paradigm. Subjects were sequentially trained to produce either wh-movement (i.e., who questions, object clefts) or NP-movement (i.e., passives, subject-raising structures) derived sentences. Throughout training, generalization to untrained sentences relying on both types of movement was tested. The influence of training on aspects of narrative discourse also was examined. Results showed generalization patterns constrained to type of movement. Training wh-movement structures resulted in generalized production of untrained wh-movement structures without influencing production of NP-movement structures. Similarly, training of NP-movement structures resulted in generalization only to other sentence types also relying on NP-movement. Aspects of sentence production in narrative contexts also was improved with treatment. These data indicate that movement to an argument (A) position as in NP-movement is distinct from movement to a non-argument (A-bar) position, required in wh-movement. The site where movement terminates in the s-structure of noncanonical sentences appears to influence sentence production. These findings show that linguistic properties of sentences influence sentence production breakdown and recovery in aphasia.


Rosmarinic acid exerts an antagonistic effect on nonalcoholic fatty liver disease by regulating the YAP1/TAZ-PPARγ/PGC-1α signaling pathway.
Rosmarinic acid (RA) is a water-soluble phenolic compound extracted from Boraginaceae and Lamiaceae. This study was designed to investigate the role and mechanism of action of RA in improving nonalcoholic fatty liver disease (NAFLD). Male SD rats maintained on a high fat diet and L02 cells stimulated with oleic acid were treated with RA. Our results showed that RA significantly reduced total cholesterol, triglycerides, low-density lipoprotein cholesterol, alanine aminotransferase, aspartate aminotransferase, and malondialdehyde levels and increased high-density lipoprotein cholesterol, superoxide dismutase and adenosine triphosphate levels both in vivo and in vitro. Hematoxylin and eosin staining and oil red O staining showed that RA had a good lipid-lowering effect and substantial protective effects on liver injury. Transmission electron microscopy and JC-1 fluorescence results showed that RA could improve mitochondrial damage in hepatocytes. Additionally, flow cytometry results indicated that RA inhibited ROS generation and apoptosis in L02 cells. The impaired hepatocytes were restored by using RA in NAFLD models characterized by down-regulating YAP1 and TAZ, meanwhile up-regulating PPARγ and PGC-1α. When YAP1 was over-expressed, RA reduced the expression of YAP1; however, the action of RA was significantly blocked by silencing YAP1. The experimental results indicated that RA markedly alleviated NAFLD by repairing mitochondrial damage and regulating the YAP1/TAZ-PPARγ/PGC-1α signaling pathway.


The role of miR-409-3p in regulation of HPV16/18-E6 mRNA in human cervical high-grade squamous intraepithelial lesions.
Cervical cancer is one of the most common malignancies in women. MicroRNAs (miRNAs) are involved in a variety of fundamental cellular processes, including carcinogenesis. The potential utilization of aberrantly expressed miRNAs as novel biomarkers in cervical cancer diagnostics is growing. We investigated miRNA expression profiles during the progression of dysplasia in cervical epithelium to identify aberrantly expressed miRNAs. High-throughput miRNA profiling of high-grade precancerous lesions identified 79 miRNAs showing significant difference in expression values compared to normal cervical epithelium. Ten selected miRNAs were subsequently measured in an independent group of samples to validate them as promising biomarkers of cervical carcinogenesis. MicroRNAs miR-10b-5p, miR-34c-5p, miR-409-3p and miR-411-5p were confirmed as downregulated, while miR-10a-5p, miR-132-3p, miR-141-5p were significantly upregulated in dysplastic cervical tissues. Further investigation revealed an inverse correlation of miR-409-3p with E6 mRNA levels in precancerous cervical lesions. Subsequent in vitro analyses showed a direct involvement of this miRNA in the regulation of E6 oncogene levels, thus confirming a potential tumor suppressor function of miR-409-3p in cervical malignancies. Hence, miR-409-3p may represent a useful early marker and a potential therapeutic target for cervical cancer.


A 500-bp region, approximately 40 kb upstream of the human CYP19 (aromatase) gene, mediates placenta-specific expression in transgenic mice.
In humans, aromatase P450 (product of CYP19 gene), which catalyzes conversion of C19 steroids to estrogens, is expressed in a number of tissues, including ovary, adipose, and syncytiotrophoblast of the placenta. The 5' untranslated regions of CYP19 mRNA transcripts in these tissues are encoded by different tissue-specific first exons, which are spliced onto a common site just upstream of the translation initiation site in exon II. In placenta, the 5' untranslated region of CYP19 mRNA transcripts is encoded by exon I.1, which lies approximately 40 kb upstream of exon II. To map genomic sequences required for placenta-specific CYP19 expression, fusion genes containing 2,400 and 501 bp of placenta-specific exon I.1 5' flanking DNA linked to the human growth hormone gene (hGH), as reporter, were introduced into transgenic mice. Expression of CYP19(I.1):hGH fusion genes containing as little as 501 bp of 5' flanking DNA was placenta-specific and developmentally regulated. Furthermore, transgene expression occurred specifically in the labyrinthine trophoblast of the mouse placenta, which contains syncytial cells that may be analogous to the human syncytiotrophoblast. We show that a relatively small segment of DNA (approximately 500 bp) >40 kb upstream of the protein coding region of a human gene is able to direct expression in an appropriate tissue- and cell-specific manner in transgenic mice. These findings suggest that 5' flanking DNA within 501 bp of exon I.1 of the human CYP19 gene contains cis-acting elements that bind placenta-specific transcription factors that are conserved between humans and mice.


Recent Progress on the Molecular Mechanism of Quality Controls Induced by Ribosome Stalling.
Accurate gene expression is a prerequisite for all cellular processes. Cells actively promote correct protein folding, which prevents the accumulation of abnormal and non-functional proteins. Translation elongation is the fundamental step in gene expression to ensure cellular functions, and abnormal translation arrest is recognized and removed by the quality controls. Recent studies demonstrated that ribosome plays crucial roles as a hub for gene regulation and quality controls. Ribosome-interacting factors are critical for the quality control mechanisms responding to abnormal translation arrest by targeting its products for degradation. Aberrant mRNAs are produced by errors in mRNA maturation steps and cause aberrant translation and are eliminated by the quality control system. In this review, we focus on recent progress on two quality controls, Ribosome-associated Quality Control (RQC) and No-Go Decay (NGD), for abnormal translational elongation. These quality controls recognize aberrant ribosome stalling and induce rapid degradation of aberrant polypeptides and mRNAs thereby maintaining protein homeostasis and preventing the protein aggregation.


PRSS8 is Downregulated and Suppresses Tumour Growth and Metastases in Hepatocellular Carcinoma.
Protease serine 8 (PRSS8), a trypsin-like serine peptidase, has been shown to function as a tumour suppressor in various malignancies. The present study aimed to investigate the expression pattern, prognostic value and the biological role of PRSS8 in human hepatocellular carcinoma (HCC). PRSS8 expression in 106 HCC surgical specimens was examined by Real-time polymerase chain reaction (PCR) and immunohistochemistry, and its clinical significance was analysed. The role of PRSS8 in cell proliferation, apoptosis and invasion were examined in vitro and in vivo. PRSS8 mRNA and protein expression were decreased in most HCC tumours from that in matched adjacent non-tumour tissues. Low intratumoral PRSS8 expression was significantly correlated with poor overall survival (OS) in patients with HCC (P = 0.001). PRSS8 expression was an independent prognostic factor for OS (hazard ratio [HR] = 1.704, P = 0.009). Furthermore, restoring PRSS8 expression in high metastatic HCCLM3 cells significantly inhibited cell proliferation and invasion. In contrast, silencing PRSS8 expression in non-metastatic HepG2 cells significantly enhanced cell growth and invasion. Moreover, our in vivo data revealed that attenuated PRSS8 expression in HepG2 cells greatly promoted tumour growth, while overexpression of PRSS8 remarkably inhibited tumour growth in an HCCLM3 xenograft model. Enhanced cell growth and invasion ability mediated by the loss of PRSS8 expression was associated with downregulation of PTEN, Bax and E-cadherin and an upregulation in Bcl-2, MMP9 and N-cadherin. Our data demonstrate that PRSS8 may serve as a tumour suppressor in HCC progression, and represent a valuable prognostic marker and potential therapeutic target for HCC.


Environmental Consortium Containing Pseudomonas and Bacillus Species Synergistically Degrades Polyethylene Terephthalate Plastic.
Plastics, such as polyethylene terephthalate (PET) from water bottles, are polluting our oceans, cities, and soils. While a number of Pseudomonas species have been described that degrade aliphatic polyesters, such as polyethylene (PE) and polyurethane (PUR), few from this genus that degrade the semiaromatic polymer PET have been reported. In this study, plastic-degrading bacteria were isolated from petroleum-polluted soils and screened for lipase activity that has been associated with PET degradation. Strains and consortia of bacteria were grown in a liquid carbon-free basal medium (LCFBM) with PET as the sole carbon source. We monitored several key physical and chemical properties, including bacterial growth and modification of the plastic surface, using scanning electron microscopy (SEM) and attenuated total reflectance-Fourier transform infrared spectroscopy (ATR-FTIR) spectroscopy. We detected by-products of hydrolysis of PET using 1H-nuclear magnetic resonance (1H NMR) analysis, consistent with the ATR-FTIR data. The full consortium of five strains containing Pseudomonas and Bacillus species grew synergistically in the presence of PET and the cleavage product bis(2-hydroxyethyl) terephthalic acid (BHET) as sole sources of carbon. Secreted enzymes extracted from the full consortium were capable of fully converting BHET to the metabolically usable monomers terephthalic acid (TPA) and ethylene glycol. Draft genomes provided evidence for mixed enzymatic capabilities between the strains for metabolic degradation of TPA and ethylene glycol, the building blocks of PET polymers, indicating cooperation and ability to cross-feed in a limited nutrient environment with PET as the sole carbon source. The use of bacterial consortia for the biodegradation of PET may provide a partial solution to widespread planetary plastic accumulation.IMPORTANCE While several research groups are utilizing purified enzymes to break down postconsumer PET to the monomers TPA and ethylene glycol to produce new PET products, here, we present a group of five soil bacteria in culture that are able to partially degrade this polymer. To date, mixed Pseudomonas spp. and Bacillus spp. biodegradation of PET has not been described, and this work highlights the possibility of using bacterial consortia to biodegrade or potentially to biorecycle PET plastic waste.


Certolizumab Pegol in Japanese Patients with Moderate to Severe Plaque Psoriasis: Effect of Demographics and Baseline Disease Characteristics on Efficacy.
We present certolizumab pegol (CZP) efficacy data across patient demographic and baseline disease characteristic subgroups from a phase 2/3 trial investigating CZP treatment in Japanese patients with moderate to severe plaque psoriasis (PSO; ClinicalTrials.gov identifier: NCT03051217). Patients were randomised 1:2:2 to placebo once every 2 weeks (Q2W), CZP 400 mg Q2W and CZP 200 mg Q2W (400 mg weeks 0, 2 and 4) for 16 weeks. Patients who achieved ≥ 50% reduction in their baseline Psoriasis Area and Severity Index (PASI 50) score at week 16 continued therapy to week 52. PASI 75/90 (75% and 90% reduction, respectively) and Physician's Global Assessment (PGA) 0/1 responder rates at weeks 16 and 52 were reported for patient demographic and baseline disease characteristic subgroups, including body mass index (BMI), PASI, disease duration and prior biologic use. Non-responder imputation was used. Of the randomised patients, 2/26 patients in the placebo group, 47/53 patients in the CZP 400 mg Q2W group and and 39/48 patients in the CZP 200 mg Q2W group completed week 52. In the subgroups evaluated, week 16 efficacy was generally maintained through week 52. At week 52, PASI 75 was achieved by 84.2, 85.7 and 80.0% of patients receiving CZP 400 mg Q2W in the low (15.0-23.7 kg/m2)/intermediate (> 23.7-27.4 kg/m2)/high (> 27.4-47.0 kg/m2) BMI subgroups, respectively, and by 77.8, 70.6 and 69.2%, respectively of patients treated with CZP 200 mg Q2W. PASI 75 at week 52 was achieved by 92.9, 75.0 and 84.2% of patients receiving CZP 400 mg Q2W in the low (12.0-18.0)/intermediate (> 18.0-27.0)/high (> 27.0-67.2) baseline PASI subgroups, respectively, and by 85.0, 58.3 and 68.8% of patients receiving CZP 200 mg Q2W, respectively. Similar responses were observed across other subgroups evaluated for both CZP doses in PASI 75/90 and PGA 0/1. Clinically meaningful improvements in signs and symptoms of PSO were maintained through week 52 for CZP dosed at 400 mg Q2W or 200 mg Q2W, across patient subgroups. In general, a numerically greater response was observed for patients receiving CZP 400 mg Q2W versus those receiving CZP 200 mg Q2W across patient subgroups. ClinicalTrials.gov identifier, NCT03051217.


Antioxidant and anti-inflammatory potential of pomegranate rind extract to ameliorate cisplatin-induced acute kidney injury.
Cisplatin is a chemotherapeutic agent, but the therapeutic utility is limited due to its dose dependent nephrotoxicity. The aim of the present study was to evaluate the nephroprotective effect of pomegranate in cisplatin-induced acute kidney injury. Wistar rats were allocated into six groups as follows: the normal control, cisplatin-induced, pomegranate rind extract treatment (50, 100 and 200 mg kg(-1)) and pomegranate rind extract per se group. All the experimental test drugs/vehicle were administered orally for a period of ten days. Intraperitoneal injection of cisplatin (8 mg kg(-1)) was administered on day 7 to all the groups except the normal control and pomegranate per se group. On day 10, cisplatin resulted in significant nephrotoxicity in Wistar rats with a drastic elevation of serum creatinine and BUN, a decline in the concentrations of GSH, MDA and superoxide dismutase (SOD), and an elevation in the TNF-α level in renal tissues. Pathological changes in renal tissues were examined by histopathology and dysfunction in mitochondria and proximal tubule cells was detected by transmission electron microscopy. The rate of apoptosis and the expression of caspase-3, Il-1β and IL-6 in rat renal tissues were detected by immunohistochemistry. The administration of pomegranate at a dose of 200 mg per kg body weight significantly (p < 0.001) ameliorates increased serum creatinine and BUN. In parallel to this, pomegranate also exhibits anti-apoptotic activity through the reduction of active caspase-3 expression in kidneys. Additionally, in-silico studies also confirmed a renoprotective effect of pomegranate. The above findings suggest that pomegranate can be used as a dietary supplement in the treatment of cisplatin-induced kidney injury by reducing apoptosis, oxidative stress and inflammation.


Non-specific recognition of histone modifications by H3K9bhb antibody.
Ketone bodies are short chain fatty acids produced in the liver during periods of limited glucose availability that provide an alternative source of energy for the brain, heart, and skeletal muscle. Beyond this classical metabolic role, β-hydroxybutyrate (BHB), is gaining recognition as a pleiotropic signaling molecule. Lysine β-hydroxybutyrylation (Kbhb) is a newly discovered post-translational modification in which BHB is covalently attached to lysine ε-amino groups. This novel protein adduct is metabolically sensitive, dependent on BHB concentration, and found on proteins in multiple intracellular compartments, including the mitochondria and nucleus. Therefore, Kbhb is hypothesized to be an important component of ketone body-regulated physiology. Kbhb on histones is proposed to be an epigenetic regulator, which links metabolic alterations to gene expression. However, we found that the widely used antibody against the β-hydroxybutyrylated lysine 9 on histone H3 (H3K9bhb) also recognizes other modification(s), which are increased by deacetylation inhibition and include likely acetylations. Therefore, caution must be used when interpreting gene regulation data acquired with the H3K9bhb antibody.


High-resolution CT-derived measures of lung density are valid indexes of interstitial lung disease.
To assess the validity of computer-assisted methods in analyzing the lung parenchyma imaged with high-resolution computed tomography (HRCT), we compared computer-derived estimates of lung density to other, more traditional, measures of parenchymal injury in 24 subjects with idiopathic pulmonary fibrosis (IPF) and 60 subjects with extensive occupational exposure to asbestos. Gray scale density histograms were constructed from the HRCT images. The gray scale histogram of both study groups was of a skewed unimodal distribution. However, compared with the asbestos-exposed subjects, the patients with IPF had a gray scale distribution that was significantly shifted to the right (greater density) and flatter. In a multivariate analysis, after controlling for age and cigarette smoking, we found that the mean and median gray scale densities were independently associated with the presence of moderate-to-severe dyspnea, a higher International Labour Office chest X-ray category, a lower forced vital capacity, and a higher concentration of macrophages and eosinophils in the bronchoalveolar lavage fluid. These factors accounted for > 70% of the variance of the mean and median gray scale densities. Interestingly, no differences in gray scale density measures were noted between patients with IPF and patients with asbestosis when these other factors were taken into account. Our results suggest that computer-derived density analysis of the lung parenchyma on the HRCT scan is a valid, clinically meaningful, and objective measure of interstitial lung disease.


The Kaiser, Hunka and Bianchini Factor Similarity Coefficients: A Cautionary Note.
Kaiser, Hunka and Bianchini (1971) have given a method to compare two matrices of factor loadings based on the same variables, but different groups of individuals. They suggest expressing the factor similarity by the elements of the rotation matrix that optimizes the sum of inner products between corresponding columns or rows of the matrices. The optimal rotation involved in this method is examined from a mathematical point of view and, on the basis of this, the method is shown to be invalid.


Evaluation of models to estimate the actual evapotranspiration of soybean crop subjected to different water deficit conditions.
Optimization of water use for irrigation will only be possible with the adjustment of management, which is directly related to the correct estimate of the actual evapotranspiration of the crop. The present study aimed to evaluate different mathematical approaches used in estimating the actual evapotranspiration of a new soybean cultivar (BRS 7581RR) subjected to different water deficit conditions. For this, the ETa estimated by the FAO56 Dual, Jensen and Hermann, AquaCrop and Ritchie models was evaluated. The experimental design was randomized with five treatments and four replicates. Irrigation management in the treatments was conducted so as to maintain different levels of water available in the soil (AW). The T1 treatment was performed applying 80 to 100% AW; in T2 treatment, the allowed variation was 60 to 80% AW; in T3 treatment, it was 40 to 60% AW; in T4, from 20 to 40% AW; and in T5, from 0 to 20% AW. The results showed that the FAO56 Dual model showed better performance in soybean ETa estimation for winter conditions in most treatments, with NSE ranging from 0.42 to 0.83. In the summer, the Jensen and Hermann model showed the best results, with NSE ranging from 0.70 to 0.94.


Search for Venous Endothelial Biomarkers Heralding Venous Thromboembolism in Space: A Qualitative Systematic Review of Terrestrial Studies.
Background: The recent discovery of a venous thrombosis in the internal jugular vein of an astronaut has highlighted the need to predict the risk of venous thromboembolism in otherwise healthy individuals (VTE) in space. Virchow's triad defines the three classic risk factors for VTE: blood stasis, hypercoagulability, and endothelial disruption/dysfunction. Among these risk factors, venous endothelial disruption/dysfunction remains incompletely understood, making it difficult to accurately predict risk, set up relevant prophylactic measures and initiate timely treatment of VTE, especially in an extreme environment. Methods: A qualitative systematic review focused on endothelial disruption/dysfunction was conducted following the guidelines produced by the Space Biomedicine Systematic Review Group, which are based on Cochrane review guidelines. We aimed to assess the venous endothelial biochemical and imaging markers that may predict increased risk of VTE during spaceflight by surveying the existing knowledge base surrounding these markers in analogous populations to astronauts on the ground. Results: Limited imaging markers related to endothelial dysfunction that were outside the bounds of routine clinical practice were identified. While multiple potential biomarkers were identified that may provide insight into the etiology of endothelial dysfunction and its link to future VTE, insufficient prospective evidence is available to formally recommend screening potential astronauts or healthy patients with any currently available novel biomarker. Conclusion: Our review highlights a critical knowledge gap regarding the role biomarkers of venous endothelial disruption have in predicting and identifying VTE. Future population-based prospective studies are required to link potential risk factors and biomarkers for venous endothelial dysfunction to occurrence of VTE.


High-voltage-activated calcium current subtypes in mouse DRG neurons adapt in a subpopulation-specific manner after nerve injury.
Changes in ion channel function and expression are characteristic of neuropathic pain. Voltage-gated calcium channels (VGCCs) are integral for neurotransmission and membrane excitability, but relatively little is known about changes in their expression after nerve injury. In this study, we investigate whether peripheral nerve ligation is followed by changes in the density and proportion of high-voltage-activated (HVA) VGCC current subtypes in dorsal root ganglion (DRG) neurons, the contribution of presynaptic N-type calcium channels in evoked excitatory postsynaptic currents (EPSCs) recorded from dorsal horn neurons in the spinal cord, and the changes in expression of mRNA encoding VGCC subunits in DRG neurons. Using C57BL/6 mice [8- to 11-wk-old males (n = 91)] for partial sciatic nerve ligation or sham surgery, we performed whole cell patch-clamp recordings on isolated DRG neurons and dorsal horn neurons and measured the expression of all VGCC subunits with RT-PCR in DRG neurons. After nerve injury, the density of P/Q-type current was reduced overall in DRG neurons. There was an increase in the percentage of N-type and a decrease in that of P/Q-type current in medium- to large-diameter neurons. No changes were found in the contribution of presynaptic N-type calcium channels in evoked EPSCs recorded from dorsal horn neurons. The α2δ-1 subunit was upregulated by 1.7-fold and γ-3, γ-2, and β-4 subunits were all downregulated 1.7-fold in injured neurons compared with sham-operated neurons. This comprehensive characterization of HVA VGCC subtypes in mouse DRG neurons after nerve injury revealed changes in N- and P/Q-type current proportions only in medium- to large-diameter neurons.


Hepatitis C RNA prevalence in a Western European organ donor pool and virus transmission by organ transplantation.
Liver disease is a common finding after organ transplantation and might in part be due to transmission of hepatitis C virus (HCV). The aim of this study was to determine the prevalence of positive results with different anti-HCV tests and HCV-RNA in a local donor pool and to clarify to what extent HCV was transmitted to organ recipients. Serum samples from 207 consecutive organ donors were analysed retrospectively with anti-HCV ELISA (2nd and 3rd generation), anti-HCV RIBA (2nd generation) and HCV polymerase chain reaction (PCR). Organ recipients at risk were identified and followed up serologically and clinically. Anti-HCV seroprevalance in organ donors was 4.3% for 2nd generation ELISA, 4.8% for 3rd generation ELISA and 1.9% for 2nd generation RIBA. HCV-PCR was positive in 1.4%. Nine organs from four RIBA-positive donors were transplanted into eight recipients of whom four became anti-HCV and PCR positive after transplantation. HCV-PCR became positive several days after transplantation whereas anti-HCV seroconversion took place after 8-9 months. Two recipients developed acute liver disease and another two showed features of mild chronic liver disease but no serious complications due to HCV infection were observed.


[Usefulness of cerebral biopsy in focal cerebral lesions in patients with human immunodeficiency virus infection].
Brain mass lesions (BML) occurred in 10% of patients infected by the human immunodeficiency virus (HIV), as expression of severe and sometimes treatable diseases. However, the management of them is not well established. We analyzed, retrospectively, 26 brain biopsies (22 estereotaxic) in patients with HIV infection and BML to know their usefulness and safety. The inclusion criteria were: Failure of the anti-Toxoplasma empirical treatment, atypical scan appearance for toxoplasmosis, or severe neurological picture. Brain biopsy yielded a diagnosis in 19 patients (73.1%): Progressive multifocal leukoencephalopathy (n = 8; 30.8%), primary central nervous system lymphoma (n = 6; 23.1%), mycobacteriosis (n = 2; 7.7%), toxoplasmosis (n = 2; 7.7%), criptococcosis (n = 1), cryptosporidiosis (n = 1) and HIV-encephalitis (n = 1). In one case there was a multiple diagnosis: mycobacteriosis, toxoplasmosis, and lymphoma. Brain biopsy results decided a change in therapy in 65.4%, the resolution or improvement of the neurological process in 30.8%, and the determination of the prognosis in 30.8%. In 8 cases (30.7%) there were biopsy complications, with secondary mortality in one. Brain biopsy of BML in HIV-infected patients is a diagnostic method with a high overall diagnostic rentability and uncommon non reversible complications, offering the possibility to prescribe a specific and potentially curative treatment.


Rapid and selective detection of recombinant human erythropoietin in human blood plasma by a sensitive optical sensor.
Recombinant human erythropoietin (rHuEPO) is an important hormone drug that is used to treat several medical conditions. It is also frequently abused by athletes as a performance enhancing agent at sporting events. The time window of the rHuEPO in blood is short. Therefore, the rapid detection of rHuEPO use/abuse at points of care and in sports requires a selective analytical method and a sensitive sensor. Herein, we present a highly selective method for the rapid detection of rHuEPO in human blood plasma by a sensitive optical sensor. rHuEPO is selectively extracted from human blood plasma by a target-specific extractor chip and converted into a biothiol by reducing its disulfide bond structure. The formed biothiol reacts with a water soluble (E)-1-((6-methoxybenzo[d]thiazole-2-yl)diazenyl)naphthalene-2,6-diolHg(ii) (BAN-Hg) optical sensor and causes its rapid decomposition. This leads to a rapid change in the sensor color from blue to pink that can be observed by the naked eye. The optical sensor was used to quantify rHuEPO in the concentration range 1 × 10-8 M to 1 × 10-12 M by UV-Vis spectroscopy. For the screening of blood plasma, an EPO-specific extractor chip was synthesized and used to selectively extract the protein from the biological matrix prior to its conversion into biothiol and quantification by the optical sensor. Since many proteins have a disulfide bond structure, the new method has strong potential for their rapid sensitive and selective detection by the BAN-Hg sensor and UV-Vis spectroscopy.


Clocking the melting transition of charge and lattice order in 1T-TaS2 with ultrafast extreme-ultraviolet angle-resolved photoemission spectroscopy.
We use time- and angle-resolved photoemission spectroscopy with sub-30-fs extreme-ultraviolet pulses to map the time- and momentum-dependent electronic structure of photoexcited 1T-TaS(2). This compound is a two-dimensional Mott insulator with charge-density wave ordering. Charge order, evidenced by splitting between occupied subbands at the Brillouin zone boundary, melts well before the lattice responds. This challenges the view of a charge-density wave caused by electron-phonon coupling and Fermi-surface nesting alone, and suggests that electronic correlations play a key role in driving charge order.


Nature-inspired entwined coiled carbon mechanical metamaterials: molecular dynamics simulations.
Entwining-induced robust natural biosystems show superior mechanical performances over their counterparts. However, the role played by topological entwinement in the mechanical properties of artificial nanohelixes remains unknown. Here, the tensile characteristics of nano-entwined carbon nanocoil (ECNC) metamaterials are explored by atomistic simulations. The simulation results show that ECNCs exhibit heterogeneous pre-stress distribution along the spiral surfaces. The predicted stretching stress-strain responses correlate with the topological nano-entwining and dimensionality. Topological analysis reveals that the collective stretching of the bond and bond angle on the inner hexagon edge of the coils characterizes both early and final elastic extensions, whereas the intermediate elasticity is exclusively attributed to the inner-edged hexagon-angular deformation. The ECNCs impart pronounced tensile stiffnesses to the native structures, surprisingly with a maximum of over 13-fold higher stiffness for one triple-helix, beyond the scalability of mechanical springs in parallel, originating from the nano-entwining mechanism and increase in bulkiness. However, the reinforcement in strengths is restricted by the elastic strain limits that are degraded in ECNCs owing to the steric hindrance effect. All metastructures show superelongation-at-break due to a successive break-vs.-arrest process. Upon plastic deformation, the localized reduction in the radii of ECNCs leads to the formation of carbyne-based networks.


A prospective study of the relationship between lower body stiffness and hamstring injury in professional Australian rules footballers.
Hamstring strains remain one of the most prevalent injuries in Australian Rules football. The authors prospectively examined the relationship between musculotendinous stiffness of the hamstring and leg stiffness with hamstring injury in professional Australian Rules footballers during the 2006 season. Higher hamstring stiffness and leg stiffness are related to noncontact, soft tissue hamstring injury risk in professional Australian Rules footballers. Case-control study; Level of evidence, 3. Unilateral hamstring stiffness and leg stiffness were assessed in 136 professional footballers in the month before the commencement of the competitive season. This information was then investigated relative to players who suffered noncontact, soft tissue hamstring injuries during either matches or training throughout the season to identify whether preseason stiffness was related to injury occurrence. Fourteen tested players recorded acute, noncontact hamstring injuries, resulting in 3.3 ± 2.8 weeks of missed match play per injury. At preseason testing, the players who ended up sustaining a hamstring injury during the season recorded significantly higher mean hamstring stiffness (11%, P = .04) and leg stiffness (5%, P = .03). When considering the injured players, the leg stiffness of the involved limb was significantly higher than the noninjured players (P = .02), whereas hamstring stiffness was significantly higher on the noninvolved limb (P = .01). Further, those players who suffered a hamstring injury were significantly older than the noninjured players (P = .01). It appears that a high bilateral hamstring stiffness and leg stiffness may be a determinant in the risk of sustaining a hamstring injury. Further, relatively lower hamstring stiffness in the involved limb of injured players appears to be associated with increased injury and may be related to a lack of strength. The information from stiffness assessment may allow medical staff to determine the hamstring risk status for individual players in team sports.


[The anxiogenic activity of phenylethylamine in the social isolation test on mice].
One-day isolation of male albino mice of SHR strain significantly increased the level of social behaviour measured by the number and total duration of active contacts (sniffings). A putative endogenous anxiogen beta-phenylethylamine (5-12.5 mg/kg) inhibited the communication like human anxiogens corazole (10 mg/kg) and caffeine (200 mg/kg). Diazepam (1 mg/kg) prevented the action of phenylethylamine, phenibut (10-50 mg/kg) exerted no effect on it. The social isolation test during studying anxiogens and anxiolytics can be performed on mice as previously on rats.


Behavior change after fentanyl testing at a safe consumption space for women in Northern Mexico: A pilot study.
Fentanyl has led to an increased number of overdose deaths in North America. Testing substances for fentanyl may be a harm reduction strategy to prevent overdose. Little is known about behavior change after fentanyl testing and the attitudes around fentanyl knowledge and testing along the US-Mexico border in the context of a safe consumption site. This was a pilot quantitative and qualitative study with 30 women who use drugs at an unsanctioned safe consumption site in Mexicali, Mexico. Women participated in a quantitative survey, a semi-structured interview, and fentanyl testing of substances. Injection behavior was observed after fentanyl testing results were provided. Qualitative data were collected to explore the meanings participants attributed to fentanyl and fentanyl testing. Half of the substances tested positive for fentanyl (n=15, 50%), and all of them were in samples of black tar heroin. Among those participants who tested positive for fentanyl, 7 (47%) subsequently used less of the intended substance, 1 did not use the intended substance, and 7 (47%) did not change their behavior (i.e., used as originally intended). In qualitative interviews, a predominant theme was a description of fentanyl as dangerous and deadly and fentanyl testing as being helpful for modifying drug use behaviors. However, participants recognized that there could be no change in behavior following a positive fentanyl test in the context of not being able to find substances free of fentanyl. We observed mixed results related to behavior change after women's intended substance for use tested positive for fentanyl. Fentanyl testing was acceptable to women, but behavior change was hampered by the inability to find substances free of fentanyl. Further research is needed to maximize the potential of fentanyl testing as a harm reduction tool especially in the context of a changing drug supply.


STI epidemiology in the global older population: emerging challenges.
This paper reviews the evidence concerning the epidemiology of sexually transmitted infections (STIs) with a particular focus on older people. Evidence from North America, Australia, China and Korea clearly shows increasing rates of many STIs in the population group aged 50 years and older. Similar changes are identified in three studies conducted in sub-Saharan Africa. There is a significant lack of detailed epidemiological data from Asia-Pacific and many African countries. There is also a lack of health education and health programmes directed towards older people. It is likely that sexual ageism is the dynamic that underpins this lack of focus and resources for older people.


Trio, a Rho Family GEF, Interacts with the Presynaptic Active Zone Proteins Piccolo and Bassoon.
Synaptic vesicles (SVs) fuse with the plasma membrane at a precise location called the presynaptic active zone (AZ). This fusion is coordinated by proteins embedded within a cytoskeletal matrix assembled at the AZ (CAZ). In the present study, we have identified a novel binding partner for the CAZ proteins Piccolo and Bassoon. This interacting protein, Trio, is a member of the Dbl family of guanine nucleotide exchange factors (GEFs) known to regulate the dynamic assembly of actin and growth factor dependent axon guidance and synaptic growth. Trio was found to interact with the C-terminal PBH 9/10 domains of Piccolo and Bassoon via its own N-terminal Spectrin repeats, a domain that is also critical for its localization to the CAZ. Moreover, our data suggest that regions within the C-terminus of Trio negatively regulate its interactions with Piccolo/Bassoon. These findings provide a mechanism for the presynaptic targeting of Trio and support a model in which Piccolo and Bassoon play a role in regulating neurotransmission through interactions with proteins, including Trio, that modulate the dynamic assembly of F-actin during cycles of synaptic vesicle exo- and endocytosis.