Release of leukotriene C4 in respiratory tract during acute viral infection.
Groups of children with wheezing during respiratory illness, children without wheezing during respiratory illness, and appropriately matched healthy children were tested for the presence and concentration of leukotriene C4 (LTC4) in nasopharyngeal secretions, employing the techniques of reverse-phase high-pressure liquid chromatography and radioimmunoassay. Although most wheezing children had LTC4 in nasopharyngeal secretions, the concentration of LTC4 among wheezing children who shed respiratory viruses was found to be consistently elevated (mean 1520 +/- 228 pg/0.1 mL) compared with values in wheezing children without evidence of viral infection (mean 709 +/- 147 pg/0.1 mL). In sharp contrast, little or no LTC4 activity was detected in healthy children (mean 106 +/- 77 pg/0.1 mL). These observations suggest that respiratory viruses are stimuli for the release of mediators of inflammation such as LTC4. Thus development of virus-induced bronchospasm may be related in part to direct mucosal cell-virus interaction and the release of pharmacologically active mediators in the respiratory tract.


Measurement of transcutaneous hemoglobin concentration by noninvasive white-light spectroscopy in infants.
To compare transcutaneously spectroscopically measured hemoglobin values with venous hemoglobin values in infants. Prospective study in healthy preterm and term infants who were breathing spontaneously. Recordings were obtained from 85 stable infants (median gestational age at measurement: 36 weeks [range: 34-43 weeks]; median body weight: 1890 g [range: 1095-4360 g]). The spectroscopic hemoglobin values were corrected for inhomogeneous distribution of hemoglobin in the tissue. The venous and spectroscopic hemoglobin values were then compared by using the Bland-Altman method, which gave an error of <5%. This pilot study could illustrate a good relation between the 2 methods for measuring hemoglobin. Larger studies are required to validate the spectroscopic method in those with conditions that affect the skin microcirculation (eg, septicemia).


Effect of Remote Ischaemic Preconditioning on Liver Injury in Patients Undergoing Major Hepatectomy for Colorectal Liver Metastasis: A Pilot Randomised Controlled Feasibility Trial.
Liver resection produces excellent long-term survival for patients with colorectal liver metastases but is associated with significant morbidity and mortality from ischaemia reperfusion injury (IRI). Remote ischaemic preconditioning (RIPC) can reduce the effect of IRI. This pilot randomised controlled trial evaluated RIPC in patients undergoing major hepatectomy at the Royal Free Hospital, London. Sixteen patients were randomised to RIPC or sham control. RIPC was induced through three 10-min cycles of alternate ischaemia and reperfusion to the leg. At baseline and immediately post-resection, transaminases and indocyanine green (ICG) clearance were measured. The RIPC group had lower ALT and AST levels immediately post-resection (ALT: 43% lower 497 ± 165 vs 889 ± 170 IU/L; p = 0.019 AST: 54% lower 408 ± 166 vs 836 ± 167 IU/L; p = 0.001) and at 24 h (ALT: 41% lower 412 ± 144 vs 698 ± 137 IU/L; p = 0.026 AST: 50% lower 316 ± 116 vs 668 ± 115 IU/L; p = 0.02). ICG clearance was reduced in controls versus RIPC immediately after resection (ICG-PDR: 11.1 ± 1.1 vs 16.5 ± 1.4%/min; p = 0.035). This pilot study shows that RIPC has potential to reduce liver injury following hepatectomy justifying a prospective RCT powered to demonstrate clinical benefits.


Chronic pain after open inguinal hernia surgery: suture fixation versus self-adhesive mesh repair.
Chronic pain following inguinal hernia repair is a complex problem. Mesh fixation with sutures may be a contributing factor to this pain. The aim of this study was to compare the incidence of chronic pain and limitation of activities of daily living following inguinal hernia repair using a sutured mesh to a self-adhesive mesh, 6 months and 1 year following surgery. All consecutive patients presenting to NHS Fife for open hernia repair between January 2009 and January 2010 were included in our analysis. A prospective survey of postoperative pain and activities of daily living was conducted at 6 months and 1 year following hernia repair. Chronic pain was assessed using the SF-36 questionnaire. The primary end points for analysis were incidence of chronic pain and limitation of activities of daily living. Overall, 132 of 215 patients completed the questionnaire, 69 in the sutured group and 63 in the self-adhesive mesh group. The need for analgesics was similar during the first 24 h after surgery. Wound infections were detected in one patient in the Lichtenstein group and two in the second group. The incidence of chronic pain was 21 and 7.9% at 6 months and 18.8 and 6.3% at 1 year (p < 0.05). Moderate and vigorous activities were found to be limited some to all of the time in nine patients (60%) in the suture fixation group and in one patient in the self-adhesive group (20%, p < 0.02). Open inguinal hernia repair with a self-adhesive mesh may lead to less chronic pain and less restriction of activities of daily living than a sutured mesh fixation.


The relationship between math anxiety and math performance: The moderating role of visuospatial working memory.
According to the processing efficiency theory (PET), math anxiety would interfere with working memory resources, negatively affecting mathematical abilities. To date, few studies have explored how the interaction between math anxiety and working memory would affect different types of math tasks, especially in primary school children. Therefore, the purpose of this study was to explore whether the interplay between math anxiety and working memory would influence performance in numerical operations (i.e., math fluency task) and mathematical reasoning (i.e., math reasoning task) in a group of primary school children (N = 202). Results showed that visuospatial working memory appeared to moderate the relationship between math anxiety and math performance when the math fluency task was considered, indicating that participants with higher levels of working memory were more negatively affected by math anxiety. No interaction effect was found for the math reasoning task in which students' scores were explained only by visuospatial working memory. The findings suggest that math anxiety and visuospatial working memory interact to influence performance in the math fluency task and that this effect may vary depending on the strategies used to complete the task. On the other hand, results on the math reasoning task showed that visuospatial working memory continues to have a positive effect on the math performance independently of math anxiety. The implications in the educational setting are discussed, pointing to the importance of monitoring and intervention studies on affective factors.


Changes in soil microbial biomass and organic C pools improve the sustainability of perennial grass and legume system under organic nutrient management.
The perennial grass-legume cropping system benefits soil because of its high biomass turnover, cover cropping nature, and different foraging behaviors. We investigated the response of soil organic carbon (SOC) pools and their stock to organic and inorganic nutrient management in the Guinea grass and legume (cowpea-Egyptian clover) cropping system. Depth-wise soil samples were collected after harvesting the Egyptian clover. Based on the ease of oxidation with chromic acid, different pools of SOC oxidizable using the Walkley-Black C method, very labile, labile, less labile, non-labile; and dissolved organic C (DOC), microbial biomass C (MBC), and total organic C (TOC) in soils were analyzed for computing several indices of SOC. After 10 years of crop cycles, FYM and NPKF nutrient management recorded greater DOC, MBC, SOC stocks, and C sequestration than the NPK. Stocks of all SOC pools and carbon management index (CMI) decreased with soil depth. A significant improvement in CMI, stratification ratio, sensitivity indices, and sustainable yield index was observed under FYM and NPKF. This grass-legume intercropping system maintained a positive carbon balance sequestered at about 0.8Mg C ha-1 after 10 years without any external input. Approximately 44-51% of the applied carbon through manure was stabilized with SOC under this cropping system. The DOC, MBC, and SOC in passive pools were identified for predicting dry fodder yield. This study concludes that the application of organics in the perennial grass-legume inter cropping system can maintain long-term sustainability, enhance the C sequestration, and offset the carbon footprint of the farm enterprises.


Protein backbone structure determination using only residual dipolar couplings from one ordering medium.
Residual dipolar couplings provide significant structural information for proteins in the solution state, which makes them attractive for the rapid determination of protein folds. Unfortunately, dipolar couplings contain inherent structural ambiguities which make them difficult to use in the absence of additional information. In this paper, we describe an approach to the construction of protein backbone folds using experimental dipolar couplings based on a bounded tree search through a structural database. We filter out false positives via an overlap similarity measure that insists that protein fragments assigned to overlapping regions of the sequence must have self-consistent structures. This allows us to determine a backbone fold (including the correct Calpha-Cbeta bond orientations) using only residual dipolar coupling data obtained from one ordering medium. We demonstrate the applicability of the method using experimental data for ubiquitin.


Antibacterial strategies inspired by the oxidative stress and response networks.
Oxidative stress arises from an imbalance between the excessive accumulation of reactive oxygen species (ROS) and a cell's capability to readily detoxify them. Although ROS are spontaneously generated during the normal oxygen respiration and metabolism, the ROS generation is usually augmented by redox-cycling agents, membrane disrupters, and bactericidal antibiotics, which contributes their antimicrobial bioactivity. It is noted that all the bacteria deploy an arsenal of inducible antioxidant defense systems to cope with the devastating effect exerted by the oxidative stress: these systems include the antioxidant effectors such as catalases and the master regulators such as OxyR. The oxidative stress response is not essential for normal growth, but critical to survive the oxidative stress conditions that the bacterial pathogens may encounter due to the host immune response and/or the antibiotic treatment. Based on these, we here define the ROS-inspired antibacterial strategies to enhance the oxidative stress of ROS generation and/or to compromise the bacterial response of ROS detoxification, by delineating the ROSgenerating antimicrobials and the core concept of the bacterial response against the oxidative stress.


Novel GH109 enzymes for bioconversion of group A red blood cells to the universal donor group O.
Glycoside hydrolases (GHs) have been employed for industrial and biotechnological purposes and often play an important role in new applications. The red blood cell (RBC) antigen system depends on the composition of oligosaccharides on the surface of erythrocytes, thus defining the ABO blood type classification. Incorrect blood transfusions may lead to fatal consequences, making the availability of the correct blood group critical. In this regard, it has been demonstrated that some GHs may be helpful in the conversion of groups A and B blood types to produce group O universal donor blood. GHs belonging to the GH109 family are of particular interest for this application due to their ability to convert blood from group A to group O. This work describes the biochemical characterisation of three novel GH109 enzymes (NAg68, NAg69 and NAg71) and the exploration of their ability to produce enzymatically converted RBCs (ECO-RBC). The three enzymes showed superior specificity on pNP-α-N-acetylgalactosamine compared to previously reported GH109 enzymes. These novel enzymes were able to act on purified antigen-A trisaccharides and produce ECO-RBC from human donor blood. NAg71 converted type A RBC to group O with increased efficiency in the presence of dextran compared to a commercially available GH109, previously used for this application.


Simvastatin enhances proliferation and pluripotent gene expression by canine bone marrow-derived mesenchymal stem cells (cBM-MSCs) in vitro.
Establishing the intervention to enhance proliferation and differentiation potential is crucial for the clinical translation of stem cell-based therapy. In this study, the effects of simvastatin on these regards were explored. Canine bone marrow-derived mesenchymal stem cells (cBM-MSCs) were treated with 4 doses of simvastatin, 0.1, 1, 10, and 100 nM. Simvastatin in low-dose range, 0.1 and 1 nM, enhanced dose-dependent cell proliferation at day 5 and 7. Exploration of the mechanisms revealed that simvastatin in low-dose range dose-dependently upregulated sets of cell cycle regulators, Cyclin D1 and Cyclin D2; proliferation marker, Ki-67; and anti-apoptotic gene; Bcl-2. Interestingly, pluripotent markers, Rex1 and Oct4, were dramatically increased upon the low-dose treatment. Contrastingly, treatment with high-dose simvastatin suppressed the expression of those genes. Thus, the results suggested beneficial effects of simvastatin on cBM-MSCs proliferation and expansion. Further study regarding differentiation potential and underlying mechanisms will accelerate the clinical application of the molecule on veterinary stem cell-based therapy.


Platelet motion near a vessel wall or thrombus surface in two-dimensional whole blood simulations.
Computational simulations using a two-dimensional lattice-Boltzmann immersed boundary method were conducted to investigate the motion of platelets near a vessel wall and close to an intravascular thrombus. Physiological volume fractions of deformable red blood cells and rigid platelet-size elliptic particles were studied under arteriolar flow conditions. Tumbling of platelets in the red-blood-cell depleted zone near the vessel walls was strongly influenced by nearby red blood cells. The thickness of the red-blood-cell depleted zone was greatly reduced near a thrombus, and platelets in this zone were pushed close to the surface of the thrombus to distances that would facilitate their cohesion to it. The distance, nature, and duration of close platelet-thrombus encounters were influenced by the porosity of the thrombus. The strong influence on platelet-thrombus encounters of red-blood-cell motion and thrombus porosity must be taken into account to understand the dynamics of platelet attachment to a growing thrombus.


A study of factors affecting blood pressure and angiotensin II in newborn infants.
Blood pressure has been measured using the Doppler ultrasonic technique in 144 infants in the first 11 days of life. Venous angiotensin II levels measured by radioimmunoassay were determined in 75 of these babies. Highly significant correlations were found between systolic blood pressure and gestational age and body weight. AII was inversely related to blood pressure and significantly related to birth weight. Babies with respiratory distress were found to be significantly hypotensive and to have markedly raised AII levels.


Dexmedetomidine abates myocardial ischemia reperfusion injury through inhibition of pyroptosis via regulation of miR-665/MEF2D/Nrf2 axis.
The current study intended to delve into the mechanisms of dexmedetomidine (Dex) in regulating myocardial pyroptosis against myocardial ischemia/reperfusion injury (MIRI). The rat MIRI models were induced by ligation/release of the coronary artery in vivo and Langendorff perfusion ex vivo. Hemodynamic parameters, infarction sizes, and histopathological changes were assessed to understand the effects of Dex on MIRI. We explored the mechanisms through functional experiments on an H9c2 cell hypoxia/reoxygenation (H/R) model. Cell viability and apoptosis were evaluated using cell counting kit 8 (CCK-8) and AV/PI dual staining respectively. The expressions of miR-665 and MEF2D mRNA were detected by qRT-PCR. Western blot was employed to determine the expression levels of pyroptosis- and signaling pathway- related proteins. The interplays between miR-665 and MEF2D were validated by Dual-luciferase reporter assays. Our findings indicated that Dex preconditioning dramatically attenuated hemodynamic derangements, infarct size, and histopathological damage in rats undergoing MIRI. Dex markedly augmented cell viability, while suppressing cell apoptosis and expressions of NLRP3, cleaved-caspase-1, ASC, GSDMD, IL-1β, and IL-18 in H9c2 cells subjected to H/R injury. MiR-665 was significantly upregulated, MEF2D and Nrf2 downregulated following H/R, whereas Dex preconditioning reversed these changes. MEF2D was validated to be a target gene of miR-665. Overexpression of miR-665 decreased the expression of MEF2D and blunted the protective effects of Dex in H9c2 cells. Moreover, the functional rescue experiment further verified that Dex regulated MEF2D/Nrf2 pathway via miR-665. In conclusion, Dex mitigates MIRI through inhibiting pyroptosis via regulating miR-665/MEF2D/Nrf2 axis.


The cyclic nucleotide specificity of eight cAMP-binding proteins in Dictyostelium discoideum is correlated into three groups.
cAMP is a mediator of inter- and intracellular events in Dictyostelium discoideum and is thought to act through specific receptors. Eight forms of cAMP-binding proteins have been described in this organism: four forms of a cell surface receptor, a cell surface and extracellular phosphodiesterase, an intracellular cAMP-dependent protein kinase (CAK), and a recently identified cAMP-binding protein (CABP1) that is present on the cell surface, in the cytoplasm, and in the nucleus. In this study we have analyzed the cyclic nucleotide specificity of these cAMP-binding proteins using 13 derivatives of cAMP with modifications in the adenine, ribose, and phosphate moiety. The results suggest that the cAMP-binding proteins belong to three groups: (i) four forms of the cell surface receptor, (ii) two forms of an intracellular receptor (CABP1 and CAK), and (iii) cell surface and extracellular phosphodiesterase. cAMP is probably bound to the surface receptors in the anti conformation in a hydrophobic cleft of the receptor with essential interactions at N6H2' and O3'. In contrast, cAMP is probably bound to CAK and CABP1 in the syn conformation with essential interactions at O2', O3', O5', and exocyclic oxygen. Finally, binding of cAMP to phosphodiesterase involves only O3' and exocyclic oxygen. The cyclic nucleotide specificity of cAMP-induced processes in D. discoideum indicates that the cell surface receptors participate in the transduction of the cAMP signal during chemotaxis and cell differentiation. Functions for CABP1 and CAK in these processes are presently elusive.


Cost and speed of locomotion for rotifers.
The hypothesis that the ciliary locomotion of rotifers is size limited and that it accounts of a significant portion of the energy budget was investigated using the genera Brachionus and Asplanchna. Speed of movement was measured among clones of different size in Brachionus, which shows little size variation through development. The same tests were done among individuals of different size within a clone of Asplanchna, which shows significant postembryonic size increase. In both cases, relative speed (body lengths per second) decreased significantly as body size increased. On this basis, and ecologically limiting size for ciliary locomotion is proposed. The actual cost of locomotion was measured for Brachionus; it is 62% of total metabolism, even though the theoretical (calculated) power requirements are well below 1% of total metabolism. Ciliary locomotion in the Rotifera thus appears to be extremely inefficient (low ratio of theoretical to actual power requirements). This hypothesis is supported indirectly by the sensitivity of speed to total metabolic rate in Brachionus: both plateau over the temperature range 20-32°C and decline in parallel outside this range. Unexpectedly high actual cost of locomotion is proposed as an important disadvantage of the Rotifera, partly offsetting the advantages accruing to them from small body size.


Cell membrane-coated nanosized active targeted drug delivery systems homing to tumor cells: A review.
Cancer is a leading cause of mortality worldwide. Cell membrane-coated nanocarriers actively targeting tumor sites are known to circumvent the limitations of conventional treatments and nanosized drug delivery systems. Cell membrane-coated nanocarriers can evade the immune system and can target tumors, thereby exhibiting a prolonged circulation time, enhancing tumor accumulation, increasing cancer therapeutic efficacy, and facilitating tumor imaging in vivo. Numerous studies have focused on cell membrane-coated nanocarriers homing to tumors. The use of these biomimetic nanocarriers in combination with photothermal or photodynamic cancer therapy have received increasing attention. This review discusses various sources of cell membranes, which have been harnessed previously in this field and highlights the mechanism underlying the targeting action of these nanocarriers and the method of their extraction, along with the applications of biomimetic cell membrane-coated nanocarriers in cancer phototherapy and diagnosis. Finally, this review discusses prospects in methods to resist cancer metastasis.


Effects of propofol and sevoflurane on social and anxiety-related behaviours in sleep-deprived rats.
Sleep disorders can profoundly affect neurological function. We investigated changes in social and anxiety-related brain functional connectivity induced by sleep deprivation, and the potential therapeutic effects of the general anaesthetics propofol and sevoflurane in rats. Twelve-week-old male Sprague-Dawley rats were subjected to sleep deprivation for 20 h per day (from 14:00 to 10:00 the next day) for 4 consecutive weeks. They were free from sleep deprivation for the remaining 4 h during which they received propofol (40 mg kg-1 i.p.) or sevoflurane (2% for 2 h) per day or no treatment. These cohorts were instrumented for EEG/EMG recordings on days 2, 14, and 28. Different cohorts were used for open field and three-chambered social behavioural tests, functional MRI, nuclear magnetic resonance spectroscopy, and positron emission tomography imaging 48 h after 4 weeks of sleep deprivation. Propofol protected against sleep deprivation-induced anxiety behaviours with more time (44.7 [8.9] s vs 24.2 [4.1] s for the sleep-deprivation controls; P<0.001) spent in the central area of the open field test and improved social preference index by 30% (all P<0.01). Compared with the sleep-deprived rats, propofol treatment enhanced overall functional connectivity by 74% (P<0.05) and overall glucose metabolism by 30% (P<0.01), and improved glutamate kinetics by 20% (P<0.05). In contrast, these effects were not found after sevoflurane treatment. Unlike sevoflurane, propofol reduced sleep deprivation-induced social and anxiety-related behaviours. Propofol might be superior to sevoflurane for patients with sleep disorders who receive anaesthesia, which should be studied in clinical studies.


Heat shock protein 60 and adjuvant arthritis: a model for T cell regulation in human arthritis.
Heat shock proteins (hsp) are highly conserved, immune-dominant microbial proteins, whose expression is increased at sites of inflammation. In the experimental model of adjuvant arthritis (AA) immune responses to hsp determine the outcome of disease. AA can be transferred with a single T cell clone specific for a sequence of mycobacterial hsp65 (Mhsp65). Immunization with whole Mhsp65 on the other hand, protects in virtually all forms of experimental arthritis, including AA. This protective effect seems the consequence of the induction of a T cell response directed against self-hsp60. A similar protective effect of self-hsp60-specific T cells seems present in patients with a spontaneous remitting form of juvenile idiopathic arthritis. Next to hsp60, other hsp have similar protective effects in arthritis, while other conserved microbial proteins lack such capacity. Nasal administration of hsp60 peptides induces IL-10-driven regulatory T cells that are highly effective in suppressing arthritis. Thus hsp60, or peptides derived from hsp60, are suitable candidates for immune therapy in chronic arthritis.


Checklist of tortoise beetles (Coleoptera, Chrysomelidae, Cassidinae) from Colombia with new data and description of a new species.
A new tortoise beetle species, Cyrtonota abrili, is described from the Antioquia and Caldas departments in Colombia. New faunistic data are provided for 87 species, including 16 new additions to the country's fauna. A checklist of the known 238 species of tortoise beetles recorded from Colombia is given.


Agonist-dependent phosphorylation of the formyl peptide receptor is regulated by the membrane proximal region of the cytoplasmic tail.
Formyl peptide receptor (FPR) is a chemoattractant G protein-coupled receptor (GPCR) involved in the innate immune response against bacteria. Receptor activation is terminated by receptor phosphorylation of two serine- and threonine-rich regions located in the distal half of the cytoplasmic tail. In this study we show that introduction of an amino acid with a bulky side chain (leucine or glutamine) adjacent to a single leucine, L320, in the membrane-proximal half of the cytoplasmic tail, significantly enhanced receptor phosphorylation, beta-arrestin1/2 translocation, and receptor endocytosis, without affecting G(i)-mediated ERK1/2 activation and release of intracellular calcium. In addition, the point mutations resulted in diminished susceptibility to trypsin, suggesting a conformation different from that of wild type FPR. Alignment of the FPR sequence with the rhodopsin sequence showed that L320 resides immediately C-terminal of an amphipathic region that in rhodopsin forms helix 8. Deletion of seven amino acids (Delta309-315) from the predicted helix 8 of FPR (G307-S319) caused reduced cell signaling as well as defects in receptor phosphorylation, beta-arrestin1/2 translocation and endocytosis. Thus, the amino acid content in the N-terminal half of the cytoplasmic tail influences the structure and desensitization of FPR.


Isotretinoin administration improves sperm production in men with infertility from oligoasthenozoospermia: a pilot study.
There is currently no effective medical therapy for men with infertility due to oligoasthenozoospermia. As men with abnormal sperm production have lower concentrations of 13-cis-retinoic acid in their testes, we hypothesized that men with infertility from oligoasthenozoospermia might have improved sperm counts when treated with isotretinoin (13-cis-retinoic acid). We conducted a single-site, single-arm, pilot study to determine the effect of therapy with isotretinoin on sperm indices in 19 infertile men with oligoasthenozoospermia. Subjects were men between 21 and 60 years of age with infertility for longer than 12 months associated with sperm concentrations below 15 million sperm/mL. All men received isotretinoin 20 mg by mouth twice daily for 20 weeks. Subjects had semen analyses, physical examinations, and laboratory tests every 4 weeks during treatment. Nineteen men enrolled in the study. Median (25th, 75th) sperm concentration increased from 2.5 (0.1, 5.9) million/mL at baseline to 3.8 (2.1, 13.0) million/mL at the end of treatment (p = 0.006). No significant changes in sperm motility were observed. There was a trend toward improved sperm morphology (p = 0.056). Six pregnancies (three spontaneous and three from intracytoplasmic sperm injection) and five births occurred during the study. Four of the births, including all three of the spontaneous pregnancies, were observed in men with improvements in sperm counts with isotretinoin therapy. Treatment was well tolerated. Isotretinoin therapy improves sperm production in some men with oligoasthenozoospermia. Additional studies of isotretinoin in men with infertility from oligoasthenozoospermia are warranted.


Changes in structure and composition of lymphocyte nuclei during mitogenic stimulation.
Nuclei of lymphocytes stimulated in vitro with concanavalin A (Con A) were classified into three morphotypes: I--unstimulated; II--partially stimulated; III--fully stimulated, lymphoblastic nuclei. During the Con A-induced change from morphotype I to III nuclear volume increased up to sixfold, due to a near 10-fold increase in the interchromatinic region. At the same time, condensed chromatin rose in volume by only about 1.5-fold and became disaggregated in to small clumps. Regressive EDTA-uranyl staining demonstrated a large increase in interchromatinic fibrillar material in morphotypes II and III. Nuclear matrices isolated from stimulated murine lymphocytes showed structures comparable to the interchromatinic region of the morphotypes. The Con A-stimulated change in nuclear structure preceded onset of DNA replication and was unaffected by hydroxyurea or cytosine arabinoside. Cycloheximide blocked the structural change, even when given 20 hr after Con A. Autoradiography after [3H]leucine showed incorporation of label in the interchromatinic region of morphotype II and III nuclei, much of which remained stable during a 48-hr chase period. Nuclear structural activation was inhibited by alpha-amanitin but a significant stable nuclear RNA fraction was not detected. We conclude that an important event in lymphocyte activation is extensive synthesis of stable proteinaceous interchromatinic matrix which may be involved in chromatin remodeling and DNA replication and/or transcription.


Effect of awake craniotomy in glioblastoma in eloquent areas (GLIOMAP): a propensity score-matched analysis of an international, multicentre, cohort study.
Awake mapping has been associated with decreased neurological deficits and increased extent of resection in eloquent glioma resections. However, its effect within clinically relevant glioblastoma subgroups remains poorly understood. We aimed to assess the benefit of this technique in subgroups of patients with glioblastomas based on age, preoperative neurological morbidity, and Karnofsky Performance Score (KPS). In this propensity score-matched analysis of an international, multicentre, cohort study (GLIOMAP), patients were recruited at four tertiary centres in Europe (Erasmus MC, Rotterdam and Haaglanden MC, The Hague, Netherlands, and UZ Leuven, Leuven, Belgium) and the USA (Brigham and Women's Hospital, Boston, MA). Patients were eligible if they were aged 18-90 years, undergoing resection, had a histopathological diagnosis of primary glioblastoma, their tumour was in an eloquent or near-eloquent location, and they had a unifocal enhancing lesion. Patients either underwent awake mapping during craniotomy, or asleep resection, as per treating physician or multidisciplinary tumour board decision. We used propensity-score matching (1:3) to match patients in the awake group with those in the asleep group to create a matched cohort, and to match patients from subgroups stratified by age (<70 years vs ≥70 years), preoperative National Institute of Health Stroke Scale (NIHSS) score (score of 0-1 vs ≥2), and preoperative KPS (90-100 vs ≤80). We used Cox proportional hazard regressions to analyse the effect of awake mapping on the primary outcomes including postoperative neurological deficits (measured by deterioration in NIHSS score at 6 week, 3 months, and 6 months postoperatively), overall survival, and progression-free survival. We used logistic regression to analyse the predictive value of awake mapping and other perioperative factors on postoperative outcomes. Between Jan 1, 2010, and Oct 31, 2020, 3919 patients were recruited, of whom 1047 with tumour resection for primary eloquent glioblastoma were included in analyses as the overall unmatched cohort. After propensity-score matching, the overall matched cohort comprised 536 patients, of whom 134 had awake craniotomies and 402 had asleep resection. In the overall matched cohort, awake craniotomy versus asleep resection resulted in fewer neurological deficits at 3 months (26 [22%] of 120 vs 107 [33%] of 323; p=0·019) and 6 months (30 [26%] of 115 vs 125 [41%] of 305; p=0·0048) postoperatively, longer overall survival (median 17·0 months [95% CI 15·0-24·0] vs 14·0 months [13·0-16·0]; p=0·00054), and longer progression-free survival (median 9·0 months [8·0-11·0] vs 7·3 months [6·0-8·8]; p=0·0060). In subgroup analyses, fewer postoperative neurological deficits occurred at 3 months and at 6 months with awake craniotomy versus asleep resection in patients younger than 70 years (3 months: 22 [21%] of 103 vs 93 [34%] of 272; p=0·016; 6 months: 24 [24%] of 101 vs 108 [42%] of 258; p=0·0014), those with an NIHSS score of 0-1 (3 months: 22 [23%] of 96 vs 97 [38%] of 254; p=0·0071; 6 months: 27 [28%] of 95 vs 115 [48%] of 239; p=0·0010), and those with a KPS of 90-100 (3 months: 17 [19%] of 88 vs 74 [35%] of 237; p=0·034; 6 months: 24 [28%] of 87 vs 101 [45%] of 223, p=0·0043). Additionally, fewer postoperative neurological deficits were seen in the awake group versus the asleep group at 3 months in patients aged 70 years and older (two [13%] of 16 vs 15 [43%] of 35; p=0·033; no difference seen at 6 months), with a NIHSS score of 2 or higher (3 months: three [13%] of 23 vs 21 [36%] of 58; p=0·040) and at 6 months in those with a KPS of 80 or lower (five [18%] of 28 vs 34 [39%] of 88; p=0·043; no difference seen at 3 months). Median overall survival was longer for the awake group than the asleep group in the subgroups younger than 70 years (19·5 months [95% CI 16·0-31·0] vs 15·0 months [13·0-17·0]; p<0·0001), an NIHSS score of 0-1 (18·0 months [16·0-31·0] vs 14·0 months [13·0-16·5]; p=0·00047), and KPS of 90-100 (19·0 months [16·0-31·0] vs 14·5 months [13·0-16·5]; p=0·00058). Median progression-free survival was also longer in the awake group than in the asleep group in patients younger than 70 years (9·3 months [95% CI 8·0-12·0] vs 7·5 months [6·5-9·0]; p=0·0061), in those with an NIHSS score of 0-1 (9·5 months [9·0-12·0] vs 8·0 months [6·5-9·0]; p=0·0035), and in those with a KPS of 90-100 (10·0 months [9·0-13·0] vs 8·0 months [7·0-9·0]; p=0·0010). No difference was seen in overall survival or progression-free survival between the awake group and the asleep group for those aged 70 years and older, with NIHSS scores of 2 or higher, or with a KPS of 80 or lower. These data might aid neurosurgeons with the assessment of their surgical strategy in individual glioblastoma patients. These findings will be validated and further explored in the SAFE trial (NCT03861299) and the PROGRAM study (NCT04708171). None.


Evoked release of endogenous amino acids from rat striatal slices and its modulation.
The release of endogenous gamma-aminobutyric acid (GABA), glutamate and aspartate stimulated by high K+ was studied by superfusing rat striatal slices. Stimulation with 5 min of 30 mM K+ was applied twice (S1 and S2) at a 20 min interval. The maximum release of GABA following stimulation was 40 (at S1) and 26 (at S2) times greater than the basal release. S1 and S2 each produced a maximum release of almost same magnitude for both glutamate and aspartate (about 2.5 times basal release). The removal of Ca2+ from the perfusion medium reduced the maximum release of these amino acids by more than 80% without affecting basal release significantly. Striatal slices were next stimulated in the same way after the addition of apomorphine or haloperidol to the perfusion medium. Apomorphine, 10 or 100 microM, reduced the K+-evoked release of GABA (by 24% at S1 and 35% at S2 with 10 microM; by 37% at S1 and 47% at S2 with 100 microM) but failed to affect the simultaneous release of glutamate and aspartate. Haloperidol (1 microM) reduced the S1-induced release of GABA but had no significant effect on the subsequent S2-induced release. The evoked release of aspartate or glutamate did not respond significantly to haloperidol, but there was a tendency to a decrease in aspartate release similar to the decrease seen for GABA release, particularly with S1. Based on these results, we discuss the property of GABA, aspartate and glutamate as neurotransmitters and the possible dopaminergic regulation of the release of these amino acids.


International feasibility study for the Women's Wellness with Type 2 Diabetes Programme (WWDP): An eHealth enabled 12-week intervention programme for midlife women with type 2 diabetes.
The current study aimed to examine feasibility of participant recruitment and retention rates for the Women's Wellness with Type 2 Diabetes program (WWDP), and to assess initial efficacy of the program in improving wellbeing outcomes. 70 midlife women with type 2 diabetes mellitus (T2DM) participated in a 12-week wellness-focused intervention, the WWDP. The WWDP involved a structured book (with participatory activities), an interactive website and nurse consultations. This study had an Australian and a UK arm. Analyses were conducted using chi-square, McNemar, paired t-test, and Wilcoxon signed-ranks tests. The attrition rate for the sample was 22.2%. Overall, significant improvement was observed in diabetes distress (DD), diabetes self-efficacy, weight, BMI, menopausal symptoms and sleep symptoms from baseline to program completion at 12 weeks. Australian participants were also more likely to meet fruit recommendation guidelines and had significant waist- and hip-circumference reductions. Good retention rates and initial efficacy findings indicated feasibility of the WWDP as a promising 12-week health and wellness program for women with T2DM. They also suggest incorporating a focus on self-efficacy and gendered information may be important in improving wellness and health outcomes related to distress and menopause.


Fast spin-echo inversion-recovery imaging versus fast T2-weighted spin-echo imaging in bone marrow abnormalities.
The purpose of this investigation is to compare a fat-suppressed T2-weighted fast spin-echo (FSE) sequence in bone marrow abnormalities with an FSE STIR sequence that recently has become commercially available. Fast spin-echo images (repetition time [TR], 3500-5000 mseconds; echo time [TE], 96-114 mseconds) and FSE STIR images (TR, 3000-5000 mseconds; TE, 32-40 mseconds; inversion time [TI], 140-150 mseconds) were compared quantitatively and qualitatively calculating signal-to-noise ratios (SNRs), contrast-to-noise ratios (CNRs), and lesion conspicuity and using a qualitative scoring system. Signal-to-noise ratio (mean +/- standard deviation) was 36.4 +/- 19.3 for the FSE and 29.0 +/- 15.9 for the FSE STIR images (P = .002). Contrast-to-noise ratio (mean +/- standard deviation) was 18.7 +/- 14.3 for the FSE and 20.3 +/- 16.0 for the FSE STIR images (P = .45). Lesion conspicuity (mean +/- standard deviation) was 1.7 +/- 1.5 for the FSE and 3.5 +/- 4.0 for the FSE STIR images (P = .025). The most important difference in the qualitative evaluation related to the better signal homogeneity on the FSE STIR images. Fast spin-echo STIR images may be preferable to FSE images with fat suppression due to better image homogeneity and lesion conspicuity.


Studies toward birth and early mammalian development in space.
Sustaining life beyond Earth on either space stations or other planets will require a clear understanding of how the space environment affects key phases of mammalian reproduction and development. Pregnancy, parturition (birth) and the early development of offspring are complex processes essential for successful reproduction and the proliferation of mammalian species. While no mammal has yet undergone birth within the space environment, studies spanning the gravity continuum from 0- to 2-g are revealing startling insights into how reproduction and development may proceed under gravitational conditions deviating from those typically experienced on Earth. In this report, I review studies of pregnant Norway rats and their offspring flown in microgravity onboard the NASA Space Shuttle throughout the period corresponding to mid- to late gestation, and analogous studies of pregnant rats exposed to hypergravity (hg) onboard the NASA Ames Research Center 24-ft centrifuge. Studies of postnatal rats flown in space or exposed to centrifugation are reviewed. Although many important questions remain unanswered, the available data suggest that numerous aspects of pregnancy, birth and early mammalian development can proceed under altered gravity conditions.


Drugs, Crime, and Aggravated Felony Deportations: Moral Panic Theory and the Legal Construction of the "Criminal Alien".
The "aggravated felony" is an oft-overlooked legal distinction that provides the basis for the removal of thousands of immigrants each year. This category's broad expansion and definitive results draw from a punitive turn in crime, drug, and immigration policy, which occurred in the 1980s and 1990s. The concept of "moral panic" is a useful tool for those who seek to understand the development of punitive responses to perceived social problems. This article revisits the original formulations of moral panic theory in order to highlight the importance of societal context in determining the symbolic salience and punitive outcomes of moral panics. The goal of the article is to evaluate the thesis that a moral panic about immigrant criminality played an important role in the development of the aggravated felony category.


Chemoimmunotherapy for relapsed/refractory and progressive 17p13-deleted chronic lymphocytic leukemia (CLL) combining pentostatin, alemtuzumab, and low-dose rituximab is effective and tolerable and limits loss of CD20 expression by circulating CLL cells.
Chronic lymphocytic leukemia/small lymphocytic lymphoma (CLL) patients with purine analog refractory disease or TP53 dysfunction still have limited treatment options and poor survival. Alemtuzumab-containing chemoimmunotherapy regimens can be effective but frequently cause serious infections. We report a Phase II trial testing the efficacy and tolerability of a short-duration regimen combining pentostatin, alemtuzumab, and low-dose high-frequency rituximab designed to decrease the risk of treatment-associated infections and to limit the loss of CD20 expression by CLL cells. The study enrolled 39 patients with progressive CLL that was either relapsed/refractory (n = 36) or previously untreated with 17p13 deletion (17p13-) (n = 3). Thirteen (33%) patients had both 17p13- and TP53 mutations predicted to be dysfunctional, and eight patients had purine analog refractory CLL without TP53 dysfunction. Twenty-six (67%) patients completed therapy, with only five (13%) patients having treatment-limiting toxicity and no treatment-related deaths. Twenty-two (56%) patients responded to treatment, with 11 (28%) complete responses (four with incomplete bone marrow recovery). Median progression-free survival was 7.2 months, time to next treatment was 9.1 months, and overall survival was 34.1 months. The majority of deaths (82%) were caused by progressive disease, including transformed diffuse large B-cell lymphoma (n = 6). Correlative studies showed that low-dose rituximab activates complement and natural killer cells without a profound and sustained decrease in expression of CD20 by circulating CLL cells. We conclude that pentostatin, alemtuzumab, and low-dose high-frequency rituximab is a tolerable and effective therapy for CLL and that low-dose rituximab therapy can activate innate immune cytotoxic mechanisms without substantially decreasing CD20 expression.


Improved normal tissue protection by proton and X-ray microchannels compared to homogeneous field irradiation.
The risk of developing normal tissue injuries often limits the radiation dose that can be applied to the tumour in radiation therapy. Microbeam Radiation Therapy (MRT), a spatially fractionated photon radiotherapy is currently tested at the European Synchrotron Radiation Facility (ESRF) to improve normal tissue protection. MRT utilizes an array of microscopically thin and nearly parallel X-ray beams that are generated by a synchrotron. At the ion microprobe SNAKE in Munich focused proton microbeams ("proton microchannels") are studied to improve normal tissue protection. Here, we comparatively investigate microbeam/microchannel irradiations with sub-millimetre X-ray versus proton beams to minimize the risk of normal tissue damage in a human skin model, in vitro. Skin tissues were irradiated with a mean dose of 2 Gy over the irradiated area either with parallel synchrotron-generated X-ray beams at the ESRF or with 20 MeV protons at SNAKE using four different irradiation modes: homogeneous field, parallel lines and microchannel applications using two different channel sizes. Normal tissue viability as determined in an MTT test was significantly higher after proton or X-ray microchannel irradiation compared to a homogeneous field irradiation. In line with these findings genetic damage, as determined by the measurement of micronuclei in keratinocytes, was significantly reduced after proton or X-ray microchannel compared to a homogeneous field irradiation. Our data show that skin irradiation using either X-ray or proton microchannels maintain a higher cell viability and DNA integrity compared to a homogeneous irradiation, and thus might improve normal tissue protection after radiation therapy.


Asymptomatic uterine retroversion at 36 weeks' gestation.
Approximately 11-19% of pregnancies begin in a retroverted uterus. If spontaneous displacement does not occur, fundal impaction may be manifest by 14-16 weeks. We present a report of persistent retroversion discovered at 36 weeks' gestation. A "pelvic mass" was misdiagnosed as posterior sacculation of the uterine wall. We include a review of the literature with regard to incidence, complications, and management of this sometimes confusing condition.


Serum vitamin D and the metabolic syndrome among osteoporotic postmenopausal female patients of a family practice clinic in Jordan.
Vitamin D deficiency and insufficiency and the metabolic syndrome are two common health issues worldwide. The association between these two health problems is subject to debate. This study aims to investigate the association between vitamin D deficiency or insufficiency and the metabolic syndrome in a sample of osteoporotic postmenopausal women attending a family practice clinic in Amman-Jordan. This was an observational cross sectional study. It was carried out in the family practice clinic in Jordan University Hospital. The study included all postmenopausal osteoporotic women attending the clinic between June 2011 and May 2012, yielding a total of 326 subjects. The association between metabolic syndrome and serum vitamin D levels was investigated. Waist circumference, body mass index, triglycerides and fasting blood sugar were significantly higher among postmenopausal women with metabolic syndrome, but HDL cholesterol was significantly lower (p<0.05). The prevalence of metabolic syndrome among all study participants was 42.9%. Triglycerides and LDL cholesterol were significantly higher among women deficiency or insufficiency (p<0.05). The prevalence of vitamin D deficiency or insufficiency was 45.7%. Among patients with metabolic syndrome, the prevalence of vitamin D deficiency or insufficiency was 50.7%. Findings of the current study suggest a lack of relationship between serum vitamin D and metabolic syndrome. However, a significant inverse relationship was found between serum vitamin D levels and both serum triglycerides and LDL levels.


Comparative Evaluation of Antimicrobial Efficacy of Fluoride-Based and Self-Assembling Peptide P11-4-based Tooth Remineralization Agents on Streptococcus mutans: A Microbiological Study.
Dental caries is a biofilm-related oral disease that continues to afflict the majority of the world's population. The disease results from an interaction between specific bacteria and dietary constituents within a biofilm known as dental plaque. Among the cariogenic microorganisms, Streptococcus mutans (S. mutans) plays pivotal role in caries-inducing processes. Evaluate and compare the antimicrobial efficacy of self-assembling peptide P11-4-based tooth remineralization agents on S. mutans. An in vitro microbiological study. The antibacterial efficacy of self-assembling peptide P11-4 gel (Group 1), fluoride enhanced hydroxyapatite gel (Group 2), acidulated phosphate fluoride gel (Group 3), chlorhexidine gluconate gel 1.0% w/w (Group 4; positive control), and normal saline (Group 5; negative control) was performed using time-kill assay over a period of 24 h and the number of S. mutans colony-forming units (CFUs) were calculated. Statistical analysis was done using Kruskal-Wallis test and Mann-Whitney post hoc Test. The level of significance was set at P < 0.05. Group 1 showed mean CFUs (× 103) of 841.33 ± 3.51, Group 2 showed 10035.33 ± 60.68, Group 3 showed 1058.00 ± 56.96, Group 4 showed 0.00 ± 0.00, and Group 5 showed mean CFUs with 15226.67 ± 96.67. The difference in the mean CFUs (× 103) between different groups was statistically significant at P < 0.001. The self-assembling peptide P11-4-based tooth remineralization agent exhibited an inhibitory influence on S. mutans and hence formation of cariogenic bacteria dominant biofilm can thus be affected by its application.


Plant reproduction: Stigma receptors regulate reactive oxygen species to establish pollination barriers.
Exciting new research highlights how stigmatic receptors purposed for recognizing self-incompatible pollen interact with the FERONIA pathway to regulate stigmatic reactive oxygen species production to enforce a barrier against self-, intra- and interspecific pollen.


Comparative effects of chronic ACE inhibition and AT1 receptor blocked losartan on cardiac hypertrophy and renal function in hypertensive patients.
The present study describes the effects of losartan and the angiotensin-converting enzyme inhibitor enalapril on blood pressure, echocardiographically calculated left ventricular mass, renal function evaluated by glomerular filtration rate and quality of life. The renin-angiotensin-aldosterone system is of importance for cardiovascular growth. There is substantial experimental documentation in animals that the angiotensin II antagonist, losartan, decreases the cardiac hypertrophy response caused by elevated arterial pressure as well as intravascular volume overload. However, data in humans is scarce. This is a 3-year, randomised, double-blind study with parallel group design in 50 patients with essential hypertension. The results show that both drugs reduced blood pressure equally effectively, and also left ventricular mass (P < 0.001). After 3 years of treatment glomerular filtration rate significantly increased with losartan (P < 0.005). Serum uric acid fell modestly although significantly, dose-dependent in losartan patients compared with an increase in enalapril patients. A fall in serum potassium from the pre-study period was observed in all patients. There was no difference between treatments in terms of patient satisfaction on quality of life. Both drugs have relatively similar hormonal and haemodynamic effect, with an excellent tolerability profile; they appear to induce comparable blood pressure falls in hypertensive patients in particular, therapy based on specific Ang II blockade may offer advantages in high risk hypertensives if left ventricular hypertrophy is present. Both enalapril and losartan, in improving the renal function attenuating the intrarenal effects of angiotensin II, might be able to reverse the pathophysiology of essential hypertensive kidney disease, and should be first-choice drugs in the treatment of essential hypertension.


Improving access to medicines by popularising generics: a study of 'India's People's Medicine' scheme in two districts of Maharashtra.
In spite of being the 'pharmacy of the world', access to essential medicines for a large majority of Indians is constrained by both physical and financial reasons. According to an estimate, medicines account for 69% of household out-of-pocket spending on health care. To make quality generic medicine affordable, India's People's Medicine Scheme (Jan Aushadhi) was launched in 2008 and then revamped and rebranded as Pradhan Mantri Bhartiya Jan Ausadhi Pariyojana (PMBJP) in 2015. The current study focuses on the availability, affordability and acceptability aspects of PMBJP essential medicines. We have used a mixed-methods approach, with the survey-based quantitative component supplemented by a qualitative component consisting of in-depth interviews (IDIs). The survey was conducted in 11 PMBJP pharmacies in Mumbai and Palghar. Data were gathered on the availability, stock-outs, price and affordability of 35 essential medicines and 2 consumables. Apart from the limited coverage of essential medicines and the significant presence of Fixed dose combinations (FDCs) in the PMBJP medicine list, the availability of surveyed essential drugs was also found to be low (47%) in PMBJP outlets. Across Mumbai and Palghar districts, around 50% and 42% of medicines were found to be out of stock for the period of 3-6 months respectively. The cost of generic medicines of PMBJP outlets for treating various conditions range from 0.01 days' wages to 0.47 days' wages for the lowest paid unskilled worker in Maharashtra. The study findings show that PMBJP's unbranded generics offer great opportunities for substantial cost savings. But, in order to fully realise the potential of this scheme, some policy actions are urgently required. First, the PMBJP drug list must include all essential drugs that feature in NLEM. Second, BPPI should procure only those drugs that pass the bioequivalence test. Third, compulsory de-branding of generics should be done in a phased manner. Fourth, PMBJP's medicine procurement and distribution policies must be reviewed to address the supply chain issues. Moreover, there is a need for major pharmaceutical policy reforms to promote generic medicines in a big way. Regulations to support mandatory generic prescribing and generic substitution by pharmacists are needed.


Structure of the gene encoding the immunodominant surface antigen on the sporozoite of the human malaria parasite Plasmodium falciparum.
The gene for the circumsporozoite (CS) protein of Plasmodium falciparum has been cloned and its nucleotide sequence determined. The gene encodes a protein of 412 amino acids as deduced from the nucleotide sequence. The protein contains 41 tandem repeats of a tetrapeptide, 37 of which are Asn-Ala-Asn-Pro and four of which are Asn-Val-Asp-Pro. Monoclonal antibodies against the CS protein of Plasmodium falciparum were inhibited from binding to the protein by synthetic peptides of the repeat sequence. The CS protein of Plasmodium falciparum and the CS protein of a simian malaria parasite, Plasmodium knowlesi, have two regions of homology, one of which is present on either side of the repeat. One region contains 12 of 13 identical amino acids. Within the nucleotide sequence of this region, 25 of 27 nucleotides are conserved. The conservation of these regions in parasites widely separated in evolution suggests that they may have a function such as binding to liver cells and may represent an invariant target for immunity.


Evaluation of the innate immune responses to influenza and live-attenuated influenza vaccine infection in primary differentiated human nasal epithelial cells.
The host innate immune response to influenza virus is a key determinant of pathogenic outcomes and long-term protective immune responses against subsequent exposures. Here, we present a direct contrast of the host responses in primary differentiated human nasal epithelial cell (hNEC) cultures following infection with either a seasonal H3N2 influenza virus (WT) or the antigenically-matched live-attenuated vaccine (LAIV) strain. Comparison of the transcriptional profiles obtained 24 and 36h post-infection showed that the magnitude of gene expression was greater in LAIV infected relative to that observed in WT infected hNEC cultures. Functional enrichment analysis revealed that the antiviral and inflammatory responses were largely driven by type III IFN induction in both WT and LAIV infected cells. However, the enrichment of biological pathways involved in the recruitment of mononuclear leukocytes, antigen-presenting cells, and T lymphocytes was uniquely observed in LAIV infected cells. These observations were reflective of the host innate immune responses observed in individuals acutely infected with influenza viruses. These findings indicate that cell-intrinsic type III IFN-mediated innate immune responses in the nasal epithelium are not only crucial for viral clearance and attenuation, but may also play an important role in the induction of protective immune responses with live-attenuated vaccines.


Oncogenic potential of the DNA replication licensing protein CDT1.
The expression of a gene, designated as Retroviral insertion site (Ris)2, was activated by retroviral DNA integration in an immortalized primitive erythroid cell line, EB-PE. Ris2 was also expressed at high levels in all human tumor cell lines analysed. Consistently, NIH3T3 fibroblasts overexpressing Ris2 formed tumors in Rag2 -/- mice when injected subcutaneously. The putative RIS2 protein shows a high sequence similarity to Xenopus CDT1, Drosophila DUP, and human CDT1, a newly identified DNA replication licensing protein, suggesting that Ris2 is a mouse homologue of CDT1. Cells overexpressing Ris2/Cdt1 exhibited a quicker entry into S phase when released from serum starvation compared to controls. Our results suggest that CDT1, an essential licensing protein for DNA replication, can function as an oncogene in mammals.


Echocardiographic markers of elevated pulmonary pressure and left ventricular diastolic dysfunction are associated with exercise intolerance in adults and adolescents with homozygous sickle cell anemia in the United States and United Kingdom.
Noninvasively assessed pulmonary pressure elevations and left ventricular (LV) diastolic dysfunction are associated with increased mortality in adults with sickle cell disease, but their relationship to exercise intolerance has not been evaluated prospectively. Echocardiography, 6-minute walk distance, hemolytic rate, and serum concentrations of ferritin and erythropoietin were evaluated in a cohort of 483 subjects with homozygous hemoglobin S in the U.S. and U.K. Walk-Treatment of Pulmonary Hypertension and Sickle Cell Disease with Sildenafil Therapy (Walk-PHaSST) study. Tricuspid regurgitation velocity, which reflects systolic pulmonary artery pressure, was 2.7 to <3.0 m/s (mean±SD, 2.8±0.1) in 26% of the subjects and ≥3.0 m/s (mean±SD, 3.4±0.4) in 11%. The LV lateral E/e' ratio, which has been shown to reflect LV filling pressure in other conditions but has not been studied in sickle cell disease, was significantly higher in the groups with tricuspid regurgitation velocity ≥2.7 m/s. Increased hemolysis (P<0.0001), LV lateral E/e' ratio (P=0.0001), blood urea nitrogen (P=0.0002), and erythropoietin (P=0.002) were independently associated with an increased tricuspid regurgitation velocity. Furthermore, female sex (P<0.0001), older age (P<0.0001), LV lateral E/e' ratio (P=0.014), and tricuspid regurgitation velocity (P=0.019) were independent predictors of a shorter 6-minute walk distance. Echocardiography-estimated elevated pulmonary artery systolic pressure and LV lateral E/e' ratio were independently associated with poor exercise capacity in a large cohort of patients with sickle cell anemia. Controlled trials investigating whether strategies to prevent or delay pulmonary hypertension and/or LV diastolic dysfunction will improve exercise capacity and long-term outcomes in sickle cell anemia should be considered. Clinical Trial Registration- URL: http://www.clinicaltrials.gov. Unique identifier: NCT00492531.


Genetic monogamy and mate choice in a pair-living primate.
In pair-living mammals, genetic monogamy is extremely rare. One possible reason is that in socially monogamous animals, mate choice can be severely constrained, increasing the risk of inbreeding or pairing with an incompatible or low-quality partner. To escape these constraints, individuals might engage in extra-pair copulations. Alternatively, inbreeding can be avoided by dispersal. However, little is known about the interactions between mating system, mate choice, and dispersal in pair-living mammals. Here we genotyped 41 wild individuals from 14 groups of coppery titi monkeys (Plecturocebus cupreus) in Peruvian Amazon using 18 microsatellite loci. Parentage analyses of 18 young revealed no cases of extra-pair paternity, indicating that the study population is mostly genetically monogamous. We did not find evidence for relatedness- or heterozygosity-based mate choice. Despite the lack of evidence for active inbreeding avoidance via mate choice, mating partners were on average not related. We further found that dispersal was not sex-biased, with both sexes dispersing opportunistically over varying distances. Our findings suggest that even opportunistic dispersal, as long as it is not constrained, can generate sufficient genetic diversity to prevent inbreeding. This, in turn, can render active inbreeding avoidance via mate choice and extra-pair copulations less necessary, helping to maintain genetic monogamy.


Nonporous titanium-oxo molecular clusters that reversibly and selectively adsorb carbon dioxide.
Nonporous titanium(IV) clusters supported by carboxylate ligands show selective sorption of carbon dioxide even in the amorphous phase. A ring-type Ti8 cluster with a permanent inner pore also adsorbs gases, but not selectively.


Synthesis and total 1H- and 13C-NMR assignment of cephem derivatives for use in ADEPT approaches.
We report the synthesis and total NMR characterization of 5-thia-1-azabicyclo-[4.2.0]oct-2-ene-2-carboxylic acid-3-[[[(4''-nitrophenoxy)carbonyl]oxy]-methyl]-8-oxo-7-[(2-thienyloxoacetyl)amino]-diphenylmethyl ester-5-dioxide (5), a new cephalosporin derivative. This compound can be used as the carrier of a wide range of drugs containing an amino group. The preparation of the intermediate product, 5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid-3-[methyl 4-(6-methoxyquinolin-8-ylamino)pentylcarbamate]-8-oxo-7-[(2-thienyloxoacetyl)amino]-diphenylmethyl ester-5-dioxide (6), as well as the synthesis of the antimalarial primaquine prodrug 5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid-3-[methyl 4-(6-methoxyquinolin-8-ylamino)pentylcarbamate]-8-oxo-7-[(2-thienyloxoacetyl)amino]- 5-dioxide (7) are also described, together with their total (1)H- and (13)C-NMR assignments.


The clinical value of MRI using single-shot echoplanar DWI to identify liver involvement in patients with advanced gastroenteropancreatic-neuroendocrine tumors (GEP-NETs), compared to FSE T2 and FFE T1 weighted image after i.v. Gd-EOB-DTPA contrast enhancement.
To assess the detection rate of liver lesions in patients with advanced gastroenteropancreatic neuroendocrine carcinomas (GEP-NETs) using echo planar (EP) DWI (diffusion weighted imaging) as compared to standard FSE T2 wi and FFE T1 wi with i.v. (Gd-EOB)-DTPA. This prospective single-institution study included 55 patients with liver involvement confirmed by GEP-NETs 1.5T MRI system, using FSE T2, EP DWI and FFE T1 with i.v. (Gd-EOB)-DTPA. The potential differences between detection rates of liver deposits using 3 different MR approaches and between groups of patients were compared. Mean number of liver deposits: FSE T2=20.7, FFE T1=25.7 and tested EP DWI=24.0. No significant difference was found in overall detection rate of liver deposits seen in 3 different techniques. A significant difference in detection rate of liver deposits was noted between male vs. female and secreting vs. non-secreting cancers. There was nearly perfect agreement between both observers, and each of the tested MRI approaches in regards to number of detected liver lesions (Cohen's kappa=0.848-1). There were no significant differences among the 3 different MRI approaches in detection rates of liver deposits. Perfect agreement with high detection rate of liver deposits provides a rationale for the use of EP DWI in follow-up studies in GEP-NET patients.


Arsenite suppresses the transcriptional activity of EVI1 through the binding to CCHC-type Zn finger domain.
Transcription factor EVI1 is essential for normal hematopoiesis in embryos but is aberrantly elevated in bone marrow cells of myelodysplastic syndrome (MDS) patients. EVI1 and its downstream GATA-2 appear to be a possible therapeutic target of MDS. Here we found that treatment of EVI1-expressing K562 cells with arsenite (As(III)) reduced the mRNA and protein levels of EVI1 and GATA-2. A gel shift assay using the nuclear extract of K562 cells showed that As(III) suppressed the DNA-binding activity of EVI1. The DNA-binding activity of the recombinant EVI1 protein was also suppressed by As(III) but was recovered by excess amounts of dithiothreitol, suggesting the involvement of cysteine residues of EVI1. Since the 7th Zn finger domain of EVI1, having a motif of CCHC, is known to be involved in DNA-binding, the synthetic peptide of 7th Zn finger domain was reacted with As(III) and subjected to MALDI-TOF-MS analysis. The results showed that As(III) binds to this peptide via three cysteine residues. As(III)-induced reduction of the DNA-binding activity of the recombinant EVI1 was abolished by the mutations of each of three cysteine residues to alanine in the 7th Zn finger domain. These results demonstrate that As(III) causes the down-regulation of EVI1 and GATA-2 by inhibiting the transcriptional activity of EVI1 through the binding to the cysteine residues of CCHC-type Zn finger domain.


Associations Between Sleep Duration, Wake-Up Time, Bedtime, and Abdominal Obesity: Results From 9559 Chinese Children Aged 7-18 Years.
To investigate the associations of sleep duration, wake-up time, bedtime, and childhood abdominal obesity, and to test whether there is a weekday/weekend difference and the potential modifying role of sex. This cross-sectional study was based on the Students' Constitution and Health Survey and included 9559 students (4840 boys and 4719 girls) aged 7-18 years (7227 aged 7-12 years, 2332 aged 13-18 years). They were divided into two groups (control group and group with abdominal obesity). The physical measurements included children and youth body height, body weight, and waist circumference (WC). A parent-report questionnaire was used to collect the information about parent characteristics as well as lifestyle and sleep patterns (sleep duration, bedtime, and wake-up time of weekdays and weekends) of children and youth. The prevalence of abdominal obesity was 30.57% and the percentages of sleep duration <9 hours/day, wake-up time before 07:00 am on weekdays and weekends, bedtime after 10:00 pm on weekends were significantly higher in the group with abdominal obesity. After adjusting for confounders, sleep duration <9 hours/day on weekends was inversely related to abdominal obesity in the overall subjects, regardless of their sex and age, while bedtime after 10:00 pm on weekends was inversely related to abdominal obesity only in the overall subjects, boys, and children aged 7-12 years. Logistic regression models in all subjects showed that shorter hours of weekends sleep duration were associated with greater risks of abdominal obesity, even after adjusting for all confounders, including wake-up time and bedtime. The adjusted odds ratios and 95% confidence intervals of abdominal obesity (with ≥10 hours/day as the reference group) for children with 9-10 hours/day, 8-9 hours/day, and <8 hours/day of weekend sleep duration were 1.23 (1.04-1.46), 1.59 (1.32-1.91) and 1.83 (1.42-2.36), respectively. Specifically, after stratification by sex and age, this phenomenon was only observed in boys and children aged 7-12 years. Sleep duration and bedtime on weekends were independently associated with the risk of childhood abdominal obesity, particularly in boys and children aged 7-12 years.


Catheter-based radiofrequency renal-nerve ablation in patients with resistant hypertension.
This review aims to describe the role and the results of catheter-based renal nerve ablation for the treatment of resistant hypertension. Despite the availability of multiple classes of orally active antihypertensive treatments, resistant hypertension remains an important public health issue in 2012 due to its prevalence and association with target-organ damage and poor prognosis. The failure of purely pharmacological approaches to treat resistant hypertension has stimulated interest in invasive device-based treatments based on old concepts. In the absence of orally active antihypertensive agents, patients with severe and complicated hypertension were widely treated by surgical denervation of the kidney until the 1960s, but this approach was associated with a high incidence of severe adverse events and a high mortality rate. A new catheter system using radiofrequency energy has been developed, allowing an endovascular approach to renal denervation and providing patients with resistant hypertension with a new therapeutic option that is less invasive than surgery and can be performed rapidly under local anaesthesia. To date, this technique has been evaluated only in open-label trials including small numbers of highly selected resistant hypertensive patients with suitable renal artery anatomy. The available evidence suggests a favourable blood pressure-lowering effect in the short term (6 months) and a low incidence of immediate local and endovascular complications. This follow-up period is, however, too short for the detection of rare or late-onset adverse events. For the time being, the benefit/risk ratio of this technique remains to be evaluated, precluding its uncontrolled and widespread use in routine practice.


Serum vitamin D deficiency as a predictor of incident non-Alzheimer dementias: a 7-year longitudinal study.
Hypovitaminosis D has been cross-sectionally associated with dementia and stroke. The objective of this longitudinal study was to determine whether serum vitamin D deficiency at baseline could predict the onset of non-Alzheimer dementias (NAD) within 7 years among older women. Forty high-functioning older women (78.4 years, 76.4/82.0; median, 25th/75th percentile) from the EPIDOS Toulouse study were divided into two groups based on vitamin D deficiency (i.e., serum 25-hydroxyvitamin D <10 ng/ml) at baseline. At the end of the 7-year follow-up period, women matching the DSM-IV but not the NINCDS-ADRDA criteria were diagnosed with NAD while those matching the NINCDS-ADRDA criteria were considered to have Alzheimer's disease (AD). Subtle cognitive impairments at baseline, cardiovascular risk factors and Parkinson's disease were used as potential confounders. NAD was reported in 6 women (82.8 years, 80.6/86.0) after 7 years of follow-up. More NAD were observed in women with vitamin D deficiency (p = 0.023). There was no between-group difference regarding the onset of AD (p = 0.332). We found an association between vitamin D deficiency at baseline and the onset of NAD (adjusted odds ratio = 19.57, p = 0.042). Conversely, vitamin D deficiency was not associated with AD (p = 0.222). Baseline vitamin D deficiency predicted the onset of NAD within 7 years among older women.


Management of massive hemobilia with angiographic embolization.
Two cases of angiographic embolization for the management of massive hemobilia due to hepatico-jejunostomy and percutaneous transhepatic biliary drainage are presented. The clinical features are described together with a short review of the literature. In addition to defining the bleeding source, angiography provides an effective and safe alternative to surgery for the control of hemobilia.


Breaking the rules: tooth whitening by means of a reducing agent.
It is widely accepted that current tooth whitening treatment effect is based on the oxidizing action of peroxides, even if the mechanism of action remains still unclear. Treatments are claimed to be safe, but several secondary effects have been described, since long application times and high concentrations are needed. A faster whitening ingredient which permits the use of lower concentrations and shorter application times could potentially overcome this problem. In this work, a different approach based on a reducing agent, sodium metabisulfite (MBS), is explored. The reaction between tannic acid (TA) with carbamide peroxide (CP), MBS, and potassium persulfate (PS), as an oxidizing agent, was monitored for 48 hours by measuring its absorbance, comparing their different whitening effects. The reduction process between TA and MBS was confirmed by cyclic voltammetry. An in vitro test was used to observe if MBS whitens also stained teeth. It is shown that MBS bleaching effect is faster and higher than CP's effect over time. PS produced a darkening effect after the 3rd hour because of the strong absorbance of the oxidation metabolite. Cyclic voltammetry showed a progressive increase in the intensity of the TA anodic peak when MBS was present, demonstrating that a reduction is taking place. In vitro, MBS showed a faster whitening performance than CP, using lower concentrations. Using a TA solution as a staining model, it was possible to show that MBS has a visible bleaching effect through a reduction reaction, faster than CP, both in solution and in vitro. Low concentrations of MBS are effective in whitening. This work shows MBS as a promising candidate to develop novel whitening treatments, which is acting by reducing mechanism instead of oxidation.


The effects of low-dose fluvastatin and valsartan combination on arterial function: a randomized clinical trial.
Ageing progressively diminishes arterial functions, even in the absence of traditional risk factors. Our aim was to explore whether age-related arterial changes in middle-aged males could be reversed using short-term, low-dose fluvastatin/valsartan combination intervention. Forty apparently healthy, middle-aged males (43.3 ± 5.8 years) were recruited in a double-blind, randomised intervention. Individuals received either 10mg fluvastatin/20mg valsartan daily or placebo over 30 days. The brachial artery flow mediated dilation (FMD), pulse wave velocity (PWV) and common carotid artery β-stiffness were assessed at baseline and after 30 days, and again 5-10 months after therapy discontinuation. Arterial function variables significantly improved after 30 days of intervention; FMD improved by 167.7% (P<0.001), PWV by 10.9% (P<0.05) and β-stiffness by 18.8% (P<0.01), whereas no changes were obtained in the placebo group. The favourable outcomes in the intervention group were accompanied by a significant decrease of high sensitivity-C reactive protein levels (1.8-fold; P<0.05). In contrast, lipids and blood pressure remained unchanged. Surprisingly, the beneficial arterial effects were still present to a substantial degree 7 months after completing intervention (remaining % of initial improvement: FMD 82.1%, PWV 69.5% and β-stiffness 68.5%), but declined substantially after 10 months. Our results indicate that age-related arterial changes, at least in middle-aged males, can be reversed. Short-term treatment with a low-dose fluvastatin/valsartan combination resulted in a large and long lasting improvement of arterial function.


TCR sequencing and cloning methods for repertoire analysis and isolation of tumor-reactive TCRs.
T cell receptor (TCR) technologies, including repertoire analyses and T cell engineering, are increasingly important in the clinical management of cellular immunity in cancer, transplantation, and other immune diseases. However, sensitive and reliable methods for repertoire analyses and TCR cloning are still lacking. Here, we report on SEQTR, a high-throughput approach to analyze human and mouse repertoires that is more sensitive, reproducible, and accurate as compared with commonly used assays, and thus more reliably captures the complexity of blood and tumor TCR repertoires. We also present a TCR cloning strategy to specifically amplify TCRs from T cell populations. Positioned downstream of single-cell or bulk TCR sequencing, it allows time- and cost-effective discovery, cloning, screening, and engineering of tumor-specific TCRs. Together, these methods will accelerate TCR repertoire analyses in discovery, translational, and clinical settings and permit fast TCR engineering for cellular therapies.


Critical Property in Relaxor-PbTiO(3) Single Crystals --- Shear Piezoelectric Response.
The shear piezoelectric behavior in relaxor-PbTiO(3) (PT) single crystals is investigated in regard to crystal phase. High levels of shear piezoelectric activity, d(15) or d(24) >2000 pC N(-1), has been observed for single domain rhombohedral (R), orthorhombic (O) and tetragonal (T) relaxor-PT crystals. The high piezoelectric response is attributed to a flattening of the Gibbs free energy at compositions proximate to the morphotropic phase boundaries, where the polarization rotation is easy with applying perpendicular electric field. The shear piezoelectric behavior of pervoskite ferroelectric crystals was discussed with respect to ferroelectric-ferroelectric phase transitions and dc bias field using phenomenological approach. The relationship between single domain shear piezoelectric response and piezoelectric activities in domain engineered configurations were given in this paper. From an application viewpoint, the temperature and ac field drive stability for shear piezoelectric responses are investigated. A temperature independent shear piezoelectric response (d(24), in the range of -50°C to O-T phase transition temperature) is thermodynamically expected and experimentally confirmed in orthorhombic relaxor-PT crystals; relatively high ac field drive stability (5 kV cm(-1)) is obtained in manganese modified relaxor-PT crystals. For all thickness shear vibration modes, the mechanical quality factor Qs are less than 50, corresponding to the facilitated polarization rotation.


Magnetic-induction hyperthermia. Results of a 5-year multi-institutional national cooperative trial in advanced cancer patients.
Nine US institutions performed 14,807 Phase I-II treatments of magnetic-induction (Magnetrode [Henry Medical Electronics, Inc., Los Angeles, CA]) hyperthermia in 1170 adults. All had advanced tumors: 20% had untreated inoperable cancer or disease progression despite surgery (10%), radiation therapy (XRT) (3%), chemotherapy (27%), or combinations (40%); 67% had pain; and 79% had reduced activity. Eighteen percent were advanced primaries, 26% were recurrent, and 56% metastatic tumors in the head and neck (7%), body wall (7%), extremity (4%), abdominal cavity (17%), pelvis (17%), lung (15%), or liver (30%); 36% were less than 5 cm and 64% greater than or equal to 5 cm. Treatments were to safe tolerance for 30 to 60 minutes for five or more treatments. Results in 960 evaluable patients were complete response 9% (1-34 months; median, 7 months), partial response 18% (1-39 months; median, 4 months), minimal response 10% (1-15 months; median, 3 months), and no change 33% (1-32 months; median, 3 months), with decreased pain in 30% and improved activity in 21%, independent of histologic type or site. Regression was dependent on treatment type and minimum temperature: heat only, 23%; heat + XRT, 60%; heat + less-than-standard XRT because of prior XRT failure, 39%, heat + intravenous (IV) chemotherapy, 28%; heat + same previously failed IV chemotherapy, 20%; heat + intraarterial (IA) chemotherapy, 28%; heat + same previously failed IA chemotherapy, 15%; heat + standard XRT + chemotherapy, 58%; heat + less-than-standard XRT + chemotherapy, 47%; less than 40 degrees C, 31%; 40 to 40.9 degrees C, 45%; 41 to 41.9 degrees C, 54%; 42 to 42.9 degrees C, 47%; 43 to 43.9 degrees C, 40%; 44 to 44.9 degrees C, 33%; 45 to 45.9 degrees C, 55%; 46 to 46.9 degrees C, 63%; greater than 47 degrees C, 100%. There were 49 (0.33%) skin burns and 2 systemic injuries (stomach ulcer at 1 month; lung fibrosis at 9 months). This trial indicates that localized hyperthermia has a significant role in palliation of human advanced solid cancer.


A variant of mirror hand: a case report.
We report a variant of mirror hand in a 3.5-year-old boy who presented with multiple fingers. The x-ray revealed an attempt at fusion of 2 hypoplastic radii, 1 ulna, and multiple fingers. The unique feature of this case is a broad radius with proximal notching suggesting failed incomplete duplication. The reasons for this duplication and duplication of the fingers appear to be different.


"Fluorescence-wavelength" label-free POCT tandem with "fluorescence-photothermal" nanobody-immunosensor for detecting BSA and β-lactoglobulin.
Two carbon dots (CDs) (λEm = 525 nm, G-CDs and λEm = 640 nm, R-CDs) were synthesized from citric acid and urea. The bovine serum albumin (BSA) responsiveness of the R-CDs was used to develop a "fluorescence-wavelength" label-free point of care testing (POCT) for the detection of the milk quality marker BSA with the detection limit (LOD) of 4.89 μg/mL for fluorescence mode and 3.38 μg/mL for wavelength mode. In addition, R-CDs were found to have hydroxyl radical (·OH)-dependent fluorescence quenching properties, and a "fluorescence-photothermal" immunosensor based on nanobodies was constructed by introducing the fluorescence signal of R-CDs@BSA and the photothermal signal of oxTMB for the detection of β-lactoglobulin (β-LG) with the LOD of 0.034 ng/mL for fluorescence mode and 0.075 ng/mL for photothermal mode. The tandem detection of POCT and immunosensor enables the simultaneous and highly sensitive detection of BSA and β-LG after only simple dilution of less than 5 µL of sample.


Complications after liquid body contouring with site-enhancing oil injections.
Liquid body contouring by intramuscular injection of site-enhancing oils (SEOs) for muscle volume enlargement has increasingly become a popular practice in bodybuilding. Its immediate and long-term health consequences remain ambiguous, and postinjection complications are sparsely documented in medical literature. We report a series of patients presenting with postinjection complications upon self-attempted liquid body contouring through injection of SEOs. Ten patients visited our departments for plastic and reconstructive surgery for variable local and systemic complications. All patients gave a history of single or multiple SEO use and presented on average nine years postinjection with variable complications. Most patients experienced painful nodular indurations and swellings in the injected target muscles. Two patients showed signs of infection including erythema, fever, and local inflammatory findings. One patient presented additionally with a severe form of nonparathyroid hypercalcemia. Four patients ultimately underwent surgical treatment. The use of SEOs for liquid body contouring is a dangerous practice and is associated with potentially systemic and life-threatening complications. Surgical treatment may be the only therapeutic option in advanced cases and should be performed timely. Guidelines on diagnostic and therapeutic measures are warranted. Educational measures should be undertaken to raise awareness among athletes and medical professionals.


It Does Take Two to Tango: An Applied Conversation Analysis of Interactions between a Psychiatrist and Service-Users Discussing Medication.
Decisions concerning psychiatric medication are complex and often involve a protracted process of trial and error. We examine three recorded meetings for power-sharing and power-taking discourse strategies employed by both the psychiatrist and mental health service-user, when discussing psychiatric medication. We identify examples of good practice, as well as missed opportunities to engage service-users in co-constructed dialogue, and highlight that participation and active involvement in decisions is not best seen as a fixed pattern, but is a complex interplay that changes both between and within interactions.


The anti-hyperalgesic activity of retigabine is mediated by KCNQ potassium channel activation.
Retigabine (N-(2-amino-4-(4-fluorobenzylamino)-phenyl) carbamic acid ethyl ester) has a broad anticonvulsant spectrum and is currently in clinical development for epilepsy. The compound has an opening effect on neuronal KCNQ channels. At higher concentrations an augmentation of gamma-aminobutyric acid (GABA) induced currents as well as a weak blocking effect on sodium and calcium currents were observed. The goal of this study was to characterise the activity of retigabine in models of acute and neuropathic pain and to investigate if the potassium channel opening effect of retigabine contributes to its activity. Retigabine was tested in mice and rats in the tail flick model of acute pain and in the nerve ligation model with tight ligation of the 5th spinal nerve (L5) using both thermal and tactile stimulation. While retigabine like gabapentin had almost no analgesic effect in mice it showed some analgesic effects in rats in the tail flick model. These effects could not be antagonised with linopirdine, a selective KCNQ potassium channel blocker, indicating a different mode of action for this activity. In L5-ligated rats retigabine significantly and dose-dependently elevated the pain threshold and prolonged the withdrawal latency after tactile and thermal stimulation, respectively. In the L5 ligation model with thermal stimulation retigabine 10 mg/kg p.o. was as effective as 100 mg/kg gabapentin or 10 mg/kg tramadol. The L5 model with tactile stimulation was used to test the role of the KCNQ potassium channel opening effect of retigabine. If retigabine 10 mg/kg p.o. was administered alone it was as effective as tramadol 10 mg/kg p.o. in elevating the pain threshold. Linopirdine (1 and 3 mg/kg i.p.) had nearly no influence on neuropathic pain response. If we administered both retigabine and linopirdine the effect of retigabine was abolished or diminished depending on the dose of linopirdine used.In summary, retigabine is effective in predictive models for neuropathic pain. The activity is comparable to tramadol and is present at lower doses compared with gabapentin. Since the anti-allodynic effect can be inhibited by linopirdine we can conclude that the potassium channel opening properties of retigabine are critically involved in its ability to reduce neuropathic pain response.


Pulmonary valve echo motion in clinical practice.
Examination of the pattern of pulmonary valve echo motion provides useful diagnostic information in a variety of clinical situations. This report describes the normal patterns and variations in pulmonary valve echo motion. It further discusses the applications and limitations of M-mode and cross-sectional echocardiography in detecting obstruction to right ventricular outflow at both the valvular and infundibular level; the wide ranging effects of increases in pulmonary artery pressure on the pulmonary valve echogram; and alterations in right ventricular compliance and volume which may combine to produce diastolic opening of the pulmonary valve. It is emphasized that the thin pliable pulmonary leaflets move in response to local alterations in pressure and flow. The patterns of pulmonary valve echo motion, therefore, although not specific for any particular clinical disorder, may provide valuable, indirect information concerning local pressure and flow characteristics and, as such, may prove extremely helpful when applied to a specific clinical situation.


Sendai virus C protein impairs both phosphorylation and dephosphorylation processes of Stat1.
Sendai virus expresses C protein that blocks interferon (IFN) signaling. We previously reported suppression of IFN-stimulated tyrosine phosphorylation of signal transducers and activators of transcription (Stats) in infected cells. However this conclusion has remained controversial. To settle it, we re-examined the effect of C protein expression on phosphorylation of Stat1 in detail. IFN-stimulated tyrosine phosphorylation of Stat1 was doubtlessly suppressed early in infection, but the suppression was incomplete, suggesting the importance of the unknown blocking mechanism that inactivates the tyrosine-phosphorylated (pY)-Stat1 generated as the signaling leak. Interestingly, the dephosphorylation process of pY-Stat1 was also impaired. These effects on both phosphorylation and dephosphorylation processes were attributable to the function of the C protein.


6-month consequences of COVID-19 in patients discharged from hospital: a cohort study.
The long-term health consequences of COVID-19 remain largely unclear. The aim of this study was to describe the long-term health consequences of patients with COVID-19 who have been discharged from hospital and investigate the associated risk factors, in particular disease severity. We did an ambidirectional cohort study of patients with confirmed COVID-19 who had been discharged from Jin Yin-tan Hospital (Wuhan, China) between Jan 7 and May 29, 2020. Patients who died before follow-up; patients for whom follow-up would be difficult because of psychotic disorders, dementia, or readmission to hospital; those who were unable to move freely due to concomitant osteoarthropathy or immobile before or after discharge due to diseases such as stroke or pulmonary embolism; those who declined to participate; those who could not be contacted; and those living outside of Wuhan or in nursing or welfare homes were all excluded. All patients were interviewed with a series of questionnaires for evaluation of symptoms and health-related quality of life, underwent physical examinations and a 6-min walking test, and received blood tests. A stratified sampling procedure was used to sample patients according to their highest seven-category scale during their hospital stay as 3, 4, and 5-6, to receive pulmonary function test, high resolution CT of the chest, and ultrasonography. Enrolled patients who had participated in the Lopinavir Trial for Suppression of SARS-CoV-2 in China received SARS-CoV-2 antibody tests. Multivariable adjusted linear or logistic regression models were used to evaluate the association between disease severity and long-term health consequences. In total, 1733 of 2469 discharged patients with COVID-19 were enrolled after 736 were excluded. Patients had a median age of 57·0 years (IQR 47·0-65·0) and 897 (52%) were male and 836 (48%) were female. The follow-up study was done from June 16 to Sept 3, 2020, and the median follow-up time after symptom onset was 186·0 days (175·0-199·0). Fatigue or muscle weakness (52%, 855 of 1654) and sleep difficulties (26%, 437 of 1655) were the most common symptoms. Anxiety or depression was reported among 23% (367 of 1616) of patients. The proportions of 6-min walking distance less than the lower limit of the normal range were 17% for those at severity scale 3, 13% for severity scale 4, and 28% for severity scale 5-6. The corresponding proportions of patients with diffusion impairment were 22% for severity scale 3, 29% for scale 4, and 56% for scale 5-6, and median CT scores were 3·0 (IQR 2·0-5·0) for severity scale 3, 4·0 (3·0-5·0) for scale 4, and 5·0 (4·0-6·0) for scale 5-6. After multivariable adjustment, patients showed an odds ratio (OR) of 1·61 (95% CI 0·80-3·25) for scale 4 versus scale 3 and 4·60 (1·85-11·48) for scale 5-6 versus scale 3 for diffusion impairment; OR 0·88 (0·66-1·17) for scale 4 versus scale 3 and OR 1·76 (1·05-2·96) for scale 5-6 versus scale 3 for anxiety or depression, and OR 0·87 (0·68-1·11) for scale 4 versus scale 3 and 2·75 (1·61-4·69) for scale 5-6 versus scale 3 for fatigue or muscle weakness. Of 94 patients with blood antibodies tested at follow-up, the seropositivity (96·2% vs 58·5%) and median titres (19·0 vs 10·0) of the neutralising antibodies were significantly lower compared with at the acute phase. 107 of 822 participants without acute kidney injury and with an estimated glomerular filtration rate (eGFR) of 90 mL/min per 1·73 m2 or more at acute phase had eGFR less than 90 mL/min per 1·73 m2 at follow-up. At 6 months after acute infection, COVID-19 survivors were mainly troubled with fatigue or muscle weakness, sleep difficulties, and anxiety or depression. Patients who were more severely ill during their hospital stay had more severe impaired pulmonary diffusion capacities and abnormal chest imaging manifestations, and are the main target population for intervention of long-term recovery. National Natural Science Foundation of China, Chinese Academy of Medical Sciences Innovation Fund for Medical Sciences, National Key Research and Development Program of China, Major Projects of National Science and Technology on New Drug Creation and Development of Pulmonary Tuberculosis, and Peking Union Medical College Foundation.


Detection and discrimination of recombinant plasmid encoding hepatitis C virus core/E1 gene based on PNA and double-stranded DNA hybridization.
Development of an electrochemical DNA biosensor for direct detection and discrimination of double-stranded plasmid (ds-Pl) without the need for denaturation of the target plasmid sample using a peptide nucleic acid (PNA) oligomer as the probe is described. This goal was achieved by modification of gold electrode with 6-mercapto-1-hexanol following monolayer self-assembly of cysteine conjugated 20-mer PNA oligomer probe, complementary to the HCV core/E1 region, which binds to ds-Pl and forms PNA/ds-Pl structure. The significant variation in differential pulse voltammetric response of methylene blue on the probe modified electrode upon contacting with complementary double-strand plasmid to form PNA/ds-Pl triplex structure is the principle of target plasmid detection. The results indicated that the reduction peak current was linear with the concentration of complementary strand in the range of 10-300 pg/μl with a detection limit of 9.5 pg/μl.


Public Health Area of Concentration: a model for integration into medical school curricula.
Calls for more public health education for medical students date back at least 150 years. In recent years, medical schools have increased their required coursework in core public health topics such as epidemiology, biostatistics, and behavioral determinants of health. Some schools have created more in-depth alternatives, including combined or concurrent master's degrees; MD/PhD programs with a public health track; certificates in public health; or complete re-envisioning of the school into an integrated medical and public health institution. In 2009 the University of Pittsburgh School of Medicine began a Public Health Area of Concentration (AOC) that provides an optional, integrated curriculum that includes key elements of research, practice, and leadership. The AOC is a partnership between two schools at the University of Pittsburgh--Medicine and Public Health--and the local county health department. The result is a program that provides mentorship and training over 4 years of education designed to mend the long historical divide between the skills and constituencies of individual and population health. In addition, the AOC is relatively easy and inexpensive to implement and is modular in nature. The Public Health AOC is a simple model for incorporating many key aspects of public health into medical education and can be duplicated by any university that is willing to create partnerships and work across boundaries.


The contribution of nitric oxide to renal vascular wall thickening in rats with L-NAME-induced hypertension.
We investigated the mechanisms of renal vascular wall thickening in a rat model of N-nitro L-arginine methyl ester (L-NAME)-induced hypertension. To separate the effects of L-NAME-induced hypertension from other effects of nitric oxide (NO) inhibition, we created two models of L-NAME-induced hypertension: both had the same blood pressure level but NO inhibition was moderate in one group (group M) and severe in the other (group S). Urinary excretion of nitrates and nitrites was lower in group S than in group M. Wall thickening and lipid deposition in renal vessels were significantly greater in group S than in groups M. Simple and multiple regression analyses indicated that renal vascular wall thickening was more strongly correlated with lipid deposition than with blood pressure. The number of vessels positive for staining with Sudan black B was negatively correlated with urinary NO excretion. Expression of fibronectin and transforming growth factor-beta was greater in the Sudan black B-positive than in the Sudan black B-negative vessels, suggesting that extracellular matrix production was increased in vessels with lipid deposition. Lipid deposition and increased production of extracellular matrix may contribute to renal vascular wall thickening in L-NAME-induced hypertension. Some mechanisms independent of hypertension play important roles in vascular wall thickening induced by NO inhibition.