Histone deacetylase inhibitors stimulate tissue-type plasminogen activator production in vascular endothelial cells.
A reduced capacity for acute tissue-type plasminogen activator (t-PA) release is likely to be associated with an impaired endogenous defense against intravascular thrombosis. Efficient approaches to pharmacologically restore a defective t-PA release have been lacking, but recent observations suggest that histone deacetylase inhibitors (HDACis) enhance t-PA production in vitro. HDACis have diverse chemical structures and different HDAC-enzyme sub-class targeting. We here compared the effects of several clinically used HDACis on t-PA production in endothelial cells. Human umbilical vein endothelial cells were exposed to a panel of 11 different HDACis and t-PA mRNA and protein levels were quantified. All HDACis dose-dependently stimulated t-PA mRNA and protein expression with similar maximal efficacy but with different potencies. Already at low concentrations, the majority of inhibitors caused significant and sustained effects on t-PA production. In addition, selected HDACis were capable of normalizing t-PA production when suppressed by the inflammatory cytokine TNF-α. We conclude that HDACis targeting classical HDAC enzymes are powerful inducers of t-PA expression in cultured endothelial cells and could be promising candidates for pharmacological modulation of endogenous fibrinolysis in man.


Allelic Imbalance Analysis in Liquid Biopsy to Monitor Locally Advanced Esophageal Cancer Patients During Treatment.
Esophageal cancer (EC) is a highly aggressive tumor, and the current monitoring procedures are partially inadequate to evaluate treatment efficacy. The aim of this study was to investigate whether allelic imbalance analysis in liquid biopsy could be used as an additional tool to monitor tumor burden in EC patients. For this purpose, circulating cell-free DNA (cfDNA) from 52 patients with a locally advanced EC, which underwent neoadjuvant treatment and resection, was analyzed. Data from four representative longitudinally followed patients are also reported. Furthermore, 17 DNAs from formalin-fixed paraffin-embedded (FFPE) tumor samples were analyzed and compared to time-matched cfDNAs. To look for allelic imbalance, which is the main genetic alteration in both EC histotypes, we used a panel of five microsatellites (MSs) and three single-nucleotide polymorphisms (SNPs) near genes described as frequently altered. The Fisher exact and Mann-Whitney U tests were used to analyze categorical and continuous data, respectively. The correlation coefficient between cfDNA and FFPE-DNA was calculated with the Pearson's correlation test. We found that the selected tumor-related alterations are present in cfDNA of both adenocarcinoma (EADC) and squamous cell carcinoma (ESCC) with similar frequencies. The only exception were the MSs, one downstream and one upstream, of SMAD4 of which the loss was only observed in EADC (26 vs. 0%, P = 0.018). More interestingly, longitudinal studies disclosed that in patients with disease progression, tumor-related alterations were present in cfDNA before overt clinical or instrumental signs of relapse. In conclusion, our data indicate that the evaluation of tumor-related gene allelic imbalance in cfDNA might be a useful tool to complement the current monitoring procedures for EC patients and to guide their management.


Neovascularization after surgical exposure of rat external jugular vein endothelium.
To investigate the neovascularization after exposure of the external jugular venous endothelium in an experimental model. The sample was composed of 60 male rats of Wistar OUT B breed provided by animal colony of the Medicine College of Juazeiro do Norte, weighing 250 to 350 g, aged 90-120 days. Randomized study in OUT B Wistar rats, open, with 60 days duration. The animals were distributed into three groups of 20 specimens and were subjected to the following: Group 1: neck incision with dissection, ligation and section of the external jugular vein. Group 2: neck incision with dissection and ligation of the external jugular vein. Group 3: cervicotomy without dissection of the external jugular vein without ligation or section. The animals were sacrificed, half of them in 30 days and the other half within 60 days. The material in block harvested from the operated site were sectioned and stained for immunohistochemistry with CD34 marker. Neovascularization occurred with level of significance when compared group 1 to group 3 at 30 days (p=0.0076) and the same occurred at 60 days (p=0.0001) (Newman-Keuls test). The group with exposure of the venous endothelium showed a significant increase of neovascularization when compared with other groups.


Brain Vital Signs in Elite Ice Hockey: Towards Characterizing Objective and Specific Neurophysiological Reference Values for Concussion Management.
Background: Prior concussion studies have shown that objective neurophysiological measures are sensitive to detecting concussive and subconcussive impairments in youth ice-hockey. These studies monitored brain vital signs at rink-side using a within-subjects design to demonstrate significant changes from pre-season baseline scans. However, practical clinical implementation must overcome inherent challenges related to any dependence on a baseline. This requires establishing the start of normative reference data sets. Methods: The current study collected specific reference data for N = 58 elite, youth, male ice-hockey players and compared these with a general reference dataset from N = 135 of males and females across the lifespan. The elite hockey players were recruited to a select training camp through CAA Hockey, a management agency for players drafted to leagues such as the National Hockey League (NHL). The statistical analysis included a test-retest comparison to establish reliability, and a multivariate analysis of covariance to evaluate differences in brain vital signs between groups with age as a covariate. Findings: Test-retest assessments for brain vital signs evoked potentials showed moderate-to-good reliability (Cronbach's Alpha > 0.7, Intraclass correlation coefficient > 0.5) in five out of six measures. The multivariate analysis of covariance showed no overall effect for group (p = 0.105), and a significant effect of age as a covariate was observed (p < 0.001). Adjusting for the effect of age, a significant difference was observed in the measure of N100 latency (p = 0.022) between elite hockey players and the heterogeneous control group. Interpretation: The findings support the concept that normative physiological data can be used in brain vital signs evaluation in athletes, and should additionally be stratified for age, skill level, and experience. These can be combined with general norms and/or individual baseline assessments where appropriate and/or possible. The current results allow for brain vital sign evaluation independent of baseline assessment, therefore enabling objective neurophysiological evaluation of concussion management and cognitive performance optimization in ice-hockey.


Negative Ion Laser Desorption/Ionization Time-of-Flight Mass Spectrometric Analysis of Small Molecules Using Graphitic Carbon Nitride Nanosheet Matrix.
Ultrathin graphitic carbon nitride (g-C3N4) nanosheets served as a novel matrix for the detection of small molecules by negative ion matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) was described for the first time. In comparison with conventional organic matrices and graphene matrix, the use of g-C3N4 nanosheet matrix showed free matrix background interference and increased signal intensity in the analysis of amino acids, nucleobases, peptides, bisphenols (BPs), and nitropolycyclic aromatic hydrocarbons (nitro-PAHs). A systematic comparison of g-C3N4 nanosheets with positive and negative ion modes revealed that mass spectra produced by g-C3N4 nanosheets in negative ion mode were featured by singly deprotonated ion without matrix interference, which was rather different from the complicated alkali metal complexes in positive ion mode. Good salt tolerance and reproducibility allowed the determination of 1-nitropyrene (1-NP) in sewage, and its corresponding detection limit was lowered to 1 pmol. In addition, the ionization mechanism of the g-C3N4 nanosheets as matrix was also discussed. The work expands its application scope of g-C3N4 nanosheets and provides an alternative approach for small molecules.


Ochratoxin production by Aspergillus species.
Ochratoxin production was tested in 172 strains representing species in sections Fumigati, Circumdati, Candidi, and Wentii of the genus Aspergillus by an immunochemical method using a monoclonal antibody preparation against ochratoxin A. Ochratoxin A was detected in Aspergillus ochraceus, A. alliaceus, A. sclerotiorum, A. sulphureus, A. albertensis, A. auricomus, and A. wentii strains. This is the first report of production of ochratoxins in the latter three species. Ochratoxin production by these species was confirmed by high-performance thin-layer chromatography and by high-performance liquid chromatography. The chemical methods also indicated the production of ochratoxin B by all of the Aspergillus strains mentioned above.


Routine Stenting of Extravesical Ureteroneocystostomy in Kidney Transplantation: A Systematic Review and Meta-analysis.
Although rare, major urologic complications (MUC) in kidney transplantation can cause significant morbidity, increased cost, and may even lead to graft loss. Ureteric stents are routinely used to prevent MUC, although complications related to their use have been reported. Here, we systematically reviewed the role of routine stenting in preventing MUC in kidney transplantation with extravesical ureteric implantation and performed a meta-analysis of 6 randomized controlled trials. A PubMed search was performed for studies on MUC and stents in kidney transplant recipients. Randomized controlled trials were shortlisted for the review following Preferred Reporting Items for Systematic Reviews and Meta-Analyses guidelines. RevMan 5 was used for statistical analysis, and outcome analysis was done with Cochran-Mantel-Haenszel test using random effect model. Six trials meeting the criteria were identified. Although stent use did not decrease the incidence of urinary leak (odds ratio [OR], 0.39; 95% CI, 0.14-1.11; P = .08) or obstruction (OR, 0.41; 95% CI, 0.13-1.24; P = .11), it was associated with a higher incidence of urinary tract infection (OR, 3.59; 95% CI, 1.33-9.75; P = .01). In the present era of extravesical ureterovesical anastomosis, routine stenting has a limited role in decreasing major urologic complications and may be associated with higher incidence of urinary tract infections.


Effects of cytotoxic immunosuppressants on tuberculin-sensitive lymphocytes in guinea pigs.
The immunosuppressive activities of two phase-specific drugs, 6-mercaptopurine (6-MP) and methotrexate, and a cycle-specific agent, cyclophosphamide, were evaluated on the lymphocytic component of established tuberculin hypersensitivity in guinea pigs. In these animals, purified protein derivative (PPD)-sensitive lymphocytes are in an intermitotic phase of their proliferative cycle. Neither phase-specific drug significantly altered either the number or functional activities of these lymphocytes. By two in vitro criteria, PPD-induced lymphoproliferation and elaboration of migration inhibition factor (MIF), the responses of lymph node cells were equivalent to sensitized controls. In addition, these agents did not deplete pools of T lymphocytes, impair responses to phytohemagglutinin (PHA), nor inhibit cutaneous reactivity if employed before sensitization. In contrast, cyclophosphamide showed broader immunosuppressive effects including significant toxicities for intermitotic lymphocytes. This drug depleted pools of T cells and markedly impaired the in vitro proliferative responses of residual lymphocytes. The latter occurred with both PHA and PPD. Suppression of PHA reactivity was a dose-dependent phenomenon but was evident even with small quantities of this alkylating agent. The suppression of antigen-induced responses was independent of the proliferative status of target lymphocytes in vivo, after a single large dose, it persisted for more than 3 wk. In total, these results indicate that the effective use of cytotoxic drugs as immunosuppressants must include consideration of both the cycle specificities of the agent and the proliferative activities of the target lymphoid population.


Web crawling of social media and related web platforms to analyze backyard poultry owners responses to the 2018-2020 Newcastle Disease (ND) outbreak in Southern California.
As social media becomes an ever-increasing staple of everyday life and a growing percentage of people turn to community driven platforms as a primary source of information, the data created from these posts can provide a new source of information from which to better understand an event in near real time. The 2018-2020 outbreak of Newcastle Disease (ND) in Southern California is the third outbreak of ND in Southern California within a 50-year time span. These outbreaks are thought to be primarily driven by non-commercial poultry (i.e. backyard and game fowl) in the region. Here we employed a commercial 'web crawling' tool between June of 2018 and July of 2020 which encompassed the majority of the outbreak in order to collect all available online mentions of 'virulent Newcastle Disease' (vND), the terminology commonly used by the California Department of Food and Agriculture (CDFA), United States Department of Agriculture (USDA), and the general public, in relation to the outbreak. A total of 2498 posts in English and Spanish were returned using a Boolean logic-based string search. While the number of posts was relatively small, their impact as measured by the number of visitors to the website and the number of people viewing the post (where provided) was much larger. Posts with negative sentiment were found to have a larger audience relative to posts with a positive sentiment. In addition, posts with negative sentiment peaked in May of 2019 which preceded the formation of the anti-depopulation group Save Our Birds (SOB). As the usage and impact of social media grows, the ability to utilize tools to analyze social media may improve both response and outreach-based strategies for various disease outbreaks including vND in Southern California which has a large non-commercial poultry population.


Brominations of steroidal hormone having alpha,beta-unsaturated ketone, 17-O-acetyltestosterone, in the presence of silver triflate.
Bromination of 17-O-acetyltestosterone (17beta-acetoxyandrost-4-en-3-one) (1) was performed with 1, 5, and 10 eq of Br2 in AcOH-Et2O at room temperature. In all cases 2alpha,6beta- (2) and 2alpha,6beta-dibromo-17beta-acetoxyandrost-4-en-3-one (3) were obtained, although the yields were dependent upon the conditions used. Bromination of compound 1 with 10 eq of Br2 in the presence of silver trifluoromethanesulfonate (silver triflate, AgOTf) at room temperature for 12 h gave 2,7alpha-dibromo- (4) and 2,4,7alpha-tribromo-17beta-acetoxy-3-hydroxy-1-methylestra-1,3,5(10)-triene-6-one (5). The formations of the products were inferred on the basis of products obtained under controlled brominations of 1 in the presence of AgOTf, and of those obtained by the brominations of compounds 9-13 also in the presence of AgOTf.


Alteration of carbon and nitrogen allocation in winter wheat under elevated ozone.
Tropospheric ozone accelerates senescence and shortens grain filling, consequently affecting the remobilization and allocation efficiency of aboveground biomass and nutrients into grains in cereal crops. This study investigated carbon (C) and nitrogen (N) concentrations repeatedly in shoot biomass during the growth period and in grain after the harvest in eighteen wheat genotypes under control and ozone treatments in open-top chambers. Season-long ozone fumigation was conducted at an average ozone concentration of 70 ppb with three additional acute ozone episodes of around 150 ppb. Although there were no significant differences in straw C and N concentrations between the two treatments, the straw C:N ratio was significantly increased after long-term ozone fumigation, and the grain C:N ratio decreased under elevated ozone without significance. Grain N concentrations increased significantly under ozone stress, whereas N yield declined significantly due to grain yield losses induced by ozone. Moreover, different indicators of N use efficiency were significantly reduced with the exception of N utilization efficiency (NUtE), indicating that elevated ozone exposure reduced the N absorption from soil and allocation from vegetative to reproductive organs. The linear regression between straw C:N ratio and productivity indicated that straw C:N was not a suitable trait for predicting wheat productivity due to the low coefficient of determination (R2). Nitrogen harvest index (NHI) was not significantly affected by ozone stress among all genotypes. However, elevated ozone concentration changed the relationship between harvest index (HI) and NHI, and the reduced regression slope between them indicated that ozone exposure significantly affected the relationship of N and biomass allocation into wheat grains. The cultivar "Jenga" showed optimal ozone tolerance due to less yield reduction and higher NUE after ozone exposure. The genotypes with higher nutrient use efficiencies are promising to cope with ozone-induced changes in nitrogen partitioning.


Hsp90 stabilizes Cdc25A and counteracts heat shock-mediated Cdc25A degradation and cell-cycle attenuation in pancreatic carcinoma cells.
Pancreas cancer cells escape most treatment options. Heat shock protein (Hsp)90 is frequently over-expressed in pancreas carcinomas and protects a number of cell-cycle regulators such as the proto-oncogene Cdc25A. We show that inhibition of Hsp90 with geldanamycin (GD) destabilizes Cdc25A independent of Chk1/2, whereas the standard drug for pancreas carcinoma treatment, gemcitabine (GEM), causes Cdc25A degradation through the activation of Chk2. Both agents applied together additively inhibit the expression of Cdc25A and the proliferation of pancreas carcinoma cells thereby demonstrating that both Cdc25A-destabilizing/degrading pathways are separated. The role of Hsp90 as stabilizer of Cdc25A in pancreas carcinoma cells is further supported by two novel synthetic inhibitors 4-tosylcyclonovobiocic acid and 7-tosylcyclonovobiocic acid and specific Hsp90AB1 (Hsp90β) shRNA. Our data show that targeting Hsp90 reduced the resistance of pancreas carcinoma cells to treatment with GEM.


Dynamic changes in neurexins' alternative splicing: role of Rho-associated protein kinases and relevance to memory formation.
The three neurexins genes (NRXN1/2/3) encode polymorphic synaptic membrane proteins that are involved in cognitive functioning. Neurexins' selectivity of function is presumably conferred through differential use of 2 promoters and 5 alternative splicing sites (SS#1/2/3/4/5). In day-old rat brain neurons grown in culture, activation (depolarization) induces reversible, calcium dependent, repression of NRXN2α SS#3 insert. The effects of depolarization on NRXN1/2/3α splicing and biochemical pathways mediating them were further studied in these neurons. NRXN1/2/3α splicing in the course of memory formation in vivo was also explored, using fear conditioning paradigm in rats in which the animals were trained to associate an aversive stimulus (electrical shock) with a neutral context (a tone), resulting in the expression of fear responses to the neutral context.In the cultured neurons depolarization induced, beside NRXN2α SS#3, repression of SS#3 and SS#4 exons in NRXN3α but not NRXN1α. The repressions were mediated by the calcium/protein kinase C/Rho-associated protein kinase (ROCK) pathway. Fear conditioning induced significant and transient repressions of the NRXN1/2/3α SS#4 exons in the rat hippocampus. ROCK inhibition prior to training attenuated the behavioral fear response, the NRXN1/2/3α splicing repressions and subsequent recovery and the levels of excitatory (PSD95) and inhibitory (gephyrin) synaptic proteins in the hippocampus. No such effects were observed in the prefrontal cortex. Significant correlations existed between the fear response and hippocampal NRXN3α and NRXN2α SS#4 inserts as well as PSD95 protein levels. Hippocampal NRXN1α SS#4 insert and gephyrin levels did not correlate with the behavioral response but were negatively correlated with each other.These results show for the first time dynamic, experience related changes in NRXN1/2/3α alternative splicing in the rat brain and a role for ROCK in them. Specific neurexins' transcripts may be involved in synaptic remodeling occurring at an intermediate (hours) time scale in the course of memory formation.


Potential Toxicity of Dissolved Metal Mixtures (Cd, Cu, Pb, Zn) to Early Life Stage White Sturgeon ( Acipenser transmontanus) in the Upper Columbia River, Washington, United States.
The Upper Columbia River (UCR) received historical releases of smelter waste resulting in elevated metal concentrations in downstream sediments. Newly hatched white sturgeon hide within the rocky substrate at the sediment-water interface in the UCR for a few weeks before swim-up. Hiding behavior could expose them to metal contaminants, and metal toxicity could contribute to population declines in white sturgeon over the past 50 years. This study evaluates whether there is a link between the toxicity of dissolved metals across the sediment-water interface in the UCR and the survival of early life stage (ELS) white sturgeon. Toxicity of dissolved metal mixtures is evaluated using a combination of previously collected laboratory and field data and recently developed metal mixture toxicity models. The laboratory data consist of individual metal (Cd, Cu, Pb, and Zn) toxicity studies with ELS white sturgeon. The field data include the chemical composition of surface and pore water samples that were collected across the sediment-water interface in the UCR. These data are used in three metal accumulation and two response models. All models predict low toxicity in surface water, whereas effects concentrations greater than 20% are predicted for 60-72% of shallow pore water samples. The flux of dissolved metals, particularly Cu, from shallow pore water to surface water likely exposes prime ELS sturgeon habitat to toxic conditions.


Prevalence of acidosis and inflammation and their association with low serum albumin in chronic kidney disease.
Low serum albumin is a strong risk factor for mortality, but its association with low serum bicarbonate and inflammation in the setting of mild to moderately decreased kidney function is uncertain. We analyzed data from 15594 subjects over the age of 20 who participated in the Third National Health and Nutrition Examination Survey (NHANES III). Glomerular filtration rate (GFR) in mL/min/1.73 m2 was estimated by the abbreviated Modification of Diet in Renal Disease (MDRD) equation using appropriately calibrated serum creatinine. The age-adjusted prevalence of hypoalbuminemia (serum albumin <3.8 g/dL) at a GFR of 90, 60, 30, and 15 mL/min/1.73 m2 was 19%, 21%, 38%, and 59%, respectively, while the age-adjusted prevalence of C-reactive protein (CRP) >or= 0.22 mg/dL was 36%, 44%, 69%, and 81%, respectively, both P trend <0.001. Age, female gender, non-Hispanic black compared with non-Hispanic white race, diabetes, hypertension, hepatitis C, urine albumin: creatinine ratio >1 g/g, dietary protein intake, dietary caloric intake, serum bicarbonate, CRP, and GFR category were all significant predictors of hypoalbuminemia on univariate analysis. On simultaneously adjusting for the above variables, hypertension, diabetes, GFR, and dietary protein and caloric intake were no longer significant independent predictors of hypoalbuminemia. The adjusted odds ratio (OR) of serum bicarbonate (by quartile) for hypoalbuminemia was 1.0 for serum bicarbonate >28 mEq/L (reference), 1.25 for 26-28 mEq/L, 1.51 for 23-25 mEq/L, and 1.54 for <or=22 meq/L. The adjusted OR of CRP for hypoalbuminemia was 1.0 for CRP < 0.22 mg/dL (reference), 2.60 for 0.22-1.0 mg/dL, and 5.56 for >1.0 mg/dL. Elevated CRP and low serum bicarbonate are independently associated with hypoalbuminemia, explaining much of the high prevalence of hypoalbuminemia in chronic kidney disease.


Childhood disability in Turkana, Kenya: Understanding how carers cope in a complex humanitarian setting.
Although the consequences of disability are magnified in humanitarian contexts, research into the difficulties of caring for children with a disability in such settings has received limited attention. Based on in-depth interviews with 31 families, key informants and focus group discussions in Turkana, Kenya, this article explores the lives of families caring for children with a range of impairments (hearing, vision, physical and intellectual) in a complex humanitarian context characterised by drought, flooding, armed conflict, poverty and historical marginalisation. The challenging environmental and social conditions of Turkana magnified not only the impact of impairment on children, but also the burden of caregiving. The remoteness of Turkana, along with the paucity and fragmentation of health, rehabilitation and social services, posed major challenges and created opportunity costs for families. Disability-related stigma isolated mothers of children with disabilities, especially, increasing their burden of care and further limiting their access to services and humanitarian programmes. In a context where social systems are already stressed, the combination of these factors compounded the vulnerabilities faced by children with disabilities and their families. The needs of children with disabilities and their carers in Turkana are not being met by either community social support systems or humanitarian aid programmes. There is an urgent need to mainstream disability into Turkana services and programmes.


Quantified growth of the human embryonic heart.
The size and growth patterns of the components of the human embryonic heart have remained largely undefined. To provide these data, three-dimensional heart models were generated from immunohistochemically stained sections of ten human embryonic hearts ranging from Carnegie stage 10 to 23. Fifty-eight key structures were annotated and volumetrically assessed. Sizes of the septal foramina and atrioventricular canal opening were also measured. The heart grows exponentially throughout embryonic development. There was consistently less left than right atrial myocardium, and less right than left ventricular myocardium. We observed a later onset of trabeculation in the left atrium compared to the right. Morphometry showed that the rightward expansion of the atrioventricular canal starts in week 5. The septal foramina are less than 0.1 mm2 and are, therefore, much smaller than postnatal septal defects. This chronological, graphical atlas of the growth patterns of cardiac components in the human embryo provides quantified references for normal heart development. Thereby, this atlas may support early detection of cardiac malformations in the foetus.This article has an associated First Person interview with the first author of the paper.


Ornithine decarboxylase, transglutaminase, diamine oxidase and total diamines and polyamines in maternal liver and kidney throughout rat pregnancy.
Ornithine decarboxylase (ODC; EC 126.96.36.199), transglutaminase (EC 126.96.36.199), diamine oxidase (DAO; EC 188.8.131.52) and total di- and poly-amines were studied in rat liver and kidney cortex throughout pregnancy. In liver, ODC activity exhibited two major peaks (4.5-5 times the control activities) on days 15 and 17. Also putrescine and spermidine increased biphasically (3-4-fold), but no variation in spermine content was observed. Transglutaminase activity showed slight variations only near the end of gestation. In kidney, ODC activity did not fluctuate significantly during pregnancy, whereas both transglutaminase activity and putrescine content showed three major increases, in very early, middle and late pregnancy. No significant variations in spermidine and spermine were observed. In both organs, DAO activity, very low or undetectable until day 10, dramatically increased (10- and 20-fold in kidney and liver respectively) in the second half of pregnancy, reaching maxima on days 16-17 and 19. The results obtained for transglutaminase, ODC and total di- and poly-amines are interpreted on the basis of hyperplastic and hypertrophic events in the liver and kidney respectively. The behaviour of DAO suggests that the enzyme plays an important role in the control of intracellular diamine concentration.


I‑BET151 inhibits osteoclastogenesis via the RANKL signaling pathway in RAW264.7 macrophages.
Excessive bone resorption mediated by osteoclasts may lead to the risk of various lytic bone diseases. In the present study, the effects of I‑BET151, a bromodomain and extra terminal domain protein inhibitor, on osteoclastogenesis in RAW264.7 cells and the underlying mechanism of this process was investigated. Cells were divided into 6 groups, including the control group, receptor activator of nuclear factor‑κB ligand (RANKL) group and 4 other groups containing RANKL and I‑BET151 at different concentrations. Tartrate‑resistant acid phosphatase (TRACP) staining was used to observe the effect of I‑BET151 on osteoclastogenesis and the number of TRACP positive multinucleated cells was calculated. Western blotting was used to evaluate the expression of tumor necrosis factor receptor associated factor (TRAF6), nuclear factor of activated T‑cells cytoplasmic 1 (NFATcl), transcription factor p65 (p65), nuclear factor of κ light polypeptide gene enhancer in B‑cells inhibitor‑α (IκB‑α), extracellular signal‑regulated kinase, Jun N‑terminal kinase (JNK) and p38. mRNA expression levels of osteoclast specific genes TRACP, matrix metalloproteinase‑9 (MMP9), cathepsin K (CtsK) and proto‑oncogene tyrosine‑protein kinase Src (c‑Src) were measured using the reverse transcription‑quantitative polymerase chain reaction (RT‑qPCR). TRACP staining results demonstrated that I‑BET151 inhibited osteoclastogenesis induced by RANKL and the inhibition was dose dependent. TRACP multinucleated positive cells were significantly decreased when treated with I‑BET151 compared with the RANKL group. The inhibitory effect on TRAF6 was significant when concentrations of 100 and 200 nM I‑BET151 were used, and NFATcl was significantly inhibited when a concentration of 200 nM was used compared with the RANKL group, in a dose-dependent manner. Nuclear translocation of p65 was significantly inhibited by I‑BET151 at all concentrations. The degradation of IκB‑α, and phosphorylation of JNK and p38 were also significantly inhibited by I‑BET151, with the exception of the expression of IκB‑α following treatment with 50 nM I‑BET151. The RT‑qPCR results revealed that osteoclast‑specific genes TRACP, MMP9, CtsK and c‑Src were all dose‑dependently inhibited by I‑BET151, except for CtsK. In conclusion, I‑BET151 may significantly suppress the osteoclastogenesis of RAW264.7 cells via the RANKL signaling pathway.


Tackling seizures in patients with Alzheimer's disease.
In past years, a possible bidirectional link between epilepsy and Alzheimer's disease (AD) has been proposed: if AD patients are more likely to develop epilepsy, people with late-onset epilepsy evidence an increased risk of dementia. Furthermore, current research suggested that subclinical epileptiform discharges may be more frequent in patients with AD and network hyperexcitability may hasten cognitive impairment. In this narrative review, the authors discuss the recent evidence linking AD and epilepsy as well as seizures semeiology and epileptiform activity observed in patients with AD. Finally, anti-seizure medications (ASMs) and therapeutic trials to tackle seizures and network hyperexcitability in this clinical scenario have been summarized. There is growing experimental evidence demonstrating a strong connection between seizures, neuronal hyperexcitability, and AD. Epilepsy in AD has shown a good response to ASMs both at the late and prodromal stages. The new generation ASMs with fewer cognitive adverse effects seem to be a preferable option. Data on the possible effects of network hyperexcitability and ASMs on AD progression are still inconclusive. Further clinical trials are mandatory to identify clear guidelines about treatment of subclinical epileptiform discharges in patients with AD without seizures.


Image-derived input function for human brain using high resolution PET imaging with [C](R)-rolipram and [C]PBR28.
The aim of this study was to test seven previously published image-input methods in state-of-the-art high resolution PET brain images. Images were obtained with a High Resolution Research Tomograph plus a resolution-recovery reconstruction algorithm using two different radioligands with different radiometabolite fractions. Three of the methods required arterial blood samples to scale the image-input, and four were blood-free methods. All seven methods were tested on twelve scans with [(11)C](R)-rolipram, which has a low radiometabolite fraction, and on nineteen scans with [(11)C]PBR28 (high radiometabolite fraction). Logan V(T) values for both blood and image inputs were calculated using the metabolite-corrected input functions. The agreement of image-derived Logan V(T) values with the reference blood-derived Logan V(T) values was quantified using a scoring system. Using the image input methods that gave the most accurate results with Logan analysis, we also performed kinetic modelling with a two-tissue compartment model. For both radioligands the highest scores were obtained with two blood-based methods, while the blood-free methods generally performed poorly. All methods gave higher scores with [(11)C](R)-rolipram, which has a lower metabolite fraction. Compartment modeling gave less reliable results, especially for the estimation of individual rate constants. OUR STUDY SHOWS THAT: 1) Image input methods that are validated for a specific tracer and a specific machine may not perform equally well in a different setting; 2) despite the use of high resolution PET images, blood samples are still necessary to obtain a reliable image input function; 3) the accuracy of image input may also vary between radioligands depending on the magnitude of the radiometabolite fraction: the higher the metabolite fraction of a given tracer (e.g., [(11)C]PBR28), the more difficult it is to obtain a reliable image-derived input function; and 4) in association with image inputs, graphical analyses should be preferred over compartmental modelling.


A practical approach to relapsed multiple myeloma.
There has been tremendous progress made in multiple myeloma in the last decade, resulting in improved overall survival for all patients, including those with high-risk disease and those ineligible for transplantation. However, despite the addition of several novel agents, unprecedented response rates, and our ability to achieve complete remission in the majority of patients, the disease remains incurable in nearly all and will require repeated therapies. With many options available to the clinician, there is no simple or ideal sequence of treatments that has been established, so the choice of relapsed therapy is based on a series of factors that include response and tolerability of prior therapies, risk status, available novel agents, aggressiveness of relapse, renal function, performance status, cost, etc. This chapter provides practical guidance in selecting relapsed therapies structured through a series of 5 questions that can inform the decision. Specific emphasis is placed on the 2 most recent novel agents, carfilzomib and pomalidomide, but agents in development are also included.


[Action of 13-cis-retinoic acid and aromatic retinoid on several cutaneous parameters in hyperseborrheic rats].
Castrated rats submitted to androgenic stimulation present hyperseborrhea with an increase in foamy cells associated with an increase in cutaneous lipids and a change in the number of epidermal cell layers. 13-cis-retinoic acid and aromatic retinoid (Ro 10-9359) were given orally during 17 days (10 mg/kg) to castrated rats which had received a testosterone implant. Cyproterone acetate was chosen as the antiandrogenic drug; it was given at doses of 10 mg/kg i.m. once every other day. As opposed to cyproterone acetate, the two substances studied had no effect on the seminal vesicle and ventral prostate. 13-cis-retinoic acid and retinoic aromatic acid increased the total number of epidermal layers: In the case of 13-cis-retinoic acid, the increase involves especially the nonnucleated cell layers, whereas with retinoid aromatic acid it involves the nucleated cell layer. The number of sebaceous glands is not changed by either products, on the other hand, the number of foamy cells is considerably decreased and the weight of cutaneous lipids is decreased as well.


A framework for estimating the fixation time of an advantageous allele in stepping-stone models.
Determining how population subdivision increases the fixation time of an advantageous allele is an important problem in evolutionary genetics as this influences many processes. Here, I lay out a framework for calculating the fixation time of a positively selected allele in a subdivided population, as a function of the number of demes present, the migration rate between them and the manner in which they are connected. Using this framework, it becomes clear that a beneficial allele's fixation time is significantly reduced through migration continuously introducing copies of the allele into a newly colonized subpopulation, increasing its frequency within these demes. The effect that migration has on allele frequency needs to be explicitly taken into account to produce a realistic estimate of fixation time. This behaviour is most prominent when demes are arranged on a two-dimensional torus, in comparison with populations where demes are arranged in a circle. This is because each subpopulation is connected to several neighbours over a torus, so that there are multiple paths that an allele can take in order to fix. As a consequence, some demes experience a greater influx and efflux of migrants than others. Analytical results are found to be very accurate when compared to stochastic simulations, and are generally robust if there are a large number of demes, or if the allele is weakly selected for.


Autoinhibition of Bruton's tyrosine kinase (Btk) and activation by soluble inositol hexakisphosphate.
Bruton's tyrosine kinase (Btk), a Tec-family tyrosine kinase, is essential for B-cell function. We present crystallographic and biochemical analyses of Btk, which together reveal molecular details of its autoinhibition and activation. Autoinhibited Btk adopts a compact conformation like that of inactive c-Src and c-Abl. A lipid-binding PH-TH module, unique to Tec kinases, acts in conjunction with the SH2 and SH3 domains to stabilize the inactive conformation. In addition to the expected activation of Btk by membranes containing phosphatidylinositol triphosphate (PIP3), we found that inositol hexakisphosphate (IP6), a soluble signaling molecule found in both animal and plant cells, also activates Btk. This activation is a consequence of a transient PH-TH dimerization induced by IP6, which promotes transphosphorylation of the kinase domains. Sequence comparisons with other Tec-family kinases suggest that activation by IP6 is unique to Btk.


Corrosion and surface modification on biocompatible metals: A review.
Corrosion prevention in biomaterials has become crucial particularly to overcome inflammation and allergic reactions caused by the biomaterials' implants towards the human body. When these metal implants contacted with fluidic environments such as bloodstream and tissue of the body, most of them became mutually highly antagonistic and subsequently promotes corrosion. Biocompatible implants are typically made up of metallic, ceramic, composite and polymers. The present paper specifically focuses on biocompatible metals which favorably used as implants such as 316L stainless steel, cobalt-chromium-molybdenum, pure titanium and titanium-based alloys. This article also takes a close look at the effect of corrosion towards the implant and human body and the mechanism to improve it. Due to this corrosion delinquent, several surface modification techniques have been used to improve the corrosion behavior of biocompatible metals such as deposition of the coating, development of passivation oxide layer and ion beam surface modification. Apart from that, surface texturing methods such as plasma spraying, chemical etching, blasting, electropolishing, and laser treatment which used to improve corrosion behavior are also discussed in detail. Introduction of surface modifications to biocompatible metals is considered as a "best solution" so far to enhanced corrosion resistance performance; besides achieving superior biocompatibility and promoting osseointegration of biocompatible metals and alloys.


Human platelets contain gelsolin. A regulator of actin filament length.
Morphologic and biochemical studies suggest that actin in human platelets polymerizes in response to various stimuli and that shortening of actin filaments can be regulated by calcium. We report that human platelets contain gelsolin, a protein of Mr 91,000 that binds reversibly to actin in the presence of calcium. Platelet gelsolin exhibits immunologic crossreactivity with rabbit macrophage gelsolin and shortens actin filaments as demonstrated by viscosity measurements and gel point determinations. Gelsolin is active in micromolar calcium concentrations and its effects upon actin filaments are reversible. Gelsolin may be a dynamic regulator of actin filament length in the human platelet.


Comparative bioavailability study following a single dose intravenous and buccal administration of remdesivir in rabbits.
As remdesivir, the first FDA-approved drug for SARS-CoV-2 infection, can be used only for hospitalized patients due to intravenous administration, there is an urgent need of effective oral antiviral formulations to be used at early stage of infection in an outpatient setting. The present paper reports on the comparative pharmacokinetics of the electrospun nanofiber remdesivir/sulfobutyl ether beta-cyclodextrin formulation after intravenous and buccal administration. It was postulated that oral transmucosal administration avoids remdesivir from metabolic transformation and intact remdesivir can be detected in plasma, but only the active metabolite GS-441524 could be experimentally detected at a significantly lower plasma level, than that provided by the intravenous route. In buccally treated animals, the metabolite GS-441524 appeared only at 1 h after treatment, while in intravenously treated animals, GS-441524 was possible to quantify even at the first time-point of blood collection. Further optimization of formulation is required to improve pharmacokinetics of remdesivir-sulfobutyl ether beta-cyclodextrin formulation upon buccal administration.


Pancreatic neuroendocrine tumor: prediction of the tumor grade using CT findings and computerized texture analysis.
Background Pancreatic neuroendocrine tumors (PNET) include heterogeneous tumors with a variable degree of inherent biologic aggressiveness represented by the histopathologic grade. Although several studies investigated the computed tomography (CT) characteristics which can predict the histopathologic grade of PNET, accurate prediction of the PNET grade by CT examination alone is still limited. Purpose To investigate the important CT findings and CT texture variables for prediction of grade of PNET. Material and Methods Sixty-six patients with pathologically confirmed PNETs (grade 1 = 45, grades 2/3 = 21) underwent preoperative contrast-enhanced CT. Two reviewers determined the presence of predefined CT findings. CT texture was also analyzed on arterial and portal phase using both two-dimensional (2D) and three-dimensional (3D) analysis. Multivariate logistic regression analysis was performed in order to identify significant predictors for tumor grade. Results Among CT findings and CT texture variables, the significant predictors for grade 2/3 tumors were an ill-defined margin (odds ratio [OR] = 7.273), lower sphericity (OR = 0.409) on arterial 2D analysis, higher skewness (OR = 1.972) and lower sphericity (OR = 0.408) on arterial 3D analysis, lower kurtosis (OR = 0.436) and lower sphericity (OR = 0.420) on portal 2D analysis, and a larger surface area (OR = 2.007) and lower sphericity (OR = 0.503) on portal 3D analysis ( P < 0.05). Diagnostic performance of texture analysis was superior to CT findings (AUC = 0.774 vs. 0.683). Conclusion CT is useful for predicting grade 2/3 PNET using not only the imaging findings including an ill-defined margin, but also the CT texture variables such as lower sphericity, higher skewness, and lower kurtosis.


The noradrenergic action in antidepressant treatments: pharmacological and clinical aspects.
Even though noradrenaline has been recognized as one of the key neurotransmitters in the pathophysiology of major depression (MD), noradrenergic compounds have been less extensively utilized in clinical practice, compared to selective serotonin reuptake inhibitors (SSRIs). The development of the first selective noradrenergic reuptake inhibitor (NRI), Reboxetine, has not substantially changed the state of the art. In addition, Atomoxetine, a relatively pure NRI used for the treatment of ADHD, has shown mixed results when administered in augmentation to depressed subjects. Through a Medline search from 2000 to 2010, the present article provides an updated overview of the main pharmacological and clinical aspects of antidepressant classes that, partially or selectively, act on the noradrenergic systems. The noradrenergic action plays an important clinical effect in different antidepressant classes, as confirmed by the efficacy of dual action antidepressants such as the serotonin noradrenaline reuptake inhibitors (SNRIs), the noradrenergic and dopaminergic reuptake inhibitor (NDRI) Bupropion, and other compounds (e.g., Mianserin, Mirtazapine), which enhance the noradrenergic transmission. In addition, many tricyclics, such as Desipramine and Nortriptyline, have prevalent noradrenergic effect. Monoamine oxidase inhibitors (MAOIs), moreover, block the breakdown of serotonin, noradrenaline, dopamine and increase the availability of these monoamines. A novel class of antidepressants--the triple reuptake inhibitors--is under development to selectively act on serotonin, noradrenaline, and dopamine. Finally, the antidepressant effect of the atypical antipsychotic Quetiapine, indicated for the treatment of bipolar depression, is likely to be related to the noradrenergic action of its metabolite Norquetiapine. Even though a pure noradrenergic action might not be sufficient to obtain a full antidepressant effect, a pronoradrenergic action represents an important element for increasing the efficacy of mixed action antidepressants. In particular, the noradrenergic action seemed to be related to the motor activity, attention, and arousal.


Supporting paediatric patients to receive radiation therapy without sedation or general anaesthetic.
Many paediatric patients experience anxiety and distress when undergoing radiation therapy and, as a result, are often anaesthetised or sedated (A/S) so that they remain still. The practice of using A/S has implications for the child, the family and the health system. Building on the article by McCoola et al. (DOI 10.1002/jmrs.705), this editorial discusses approaches to improving paediatric patients' and their families' experiences of radiation therapy by reducing the need for A/S. Interventions need to be underpinned by theory and adopt robust research methods.


[Intestinal cryptosporidiosis at an early age and its negative consequences].
This review calls the attention of physicians, primarily pediatricians, to cryptosporidiosis, a still little known intestinal infection caused by the protozoan pathogen Cryptosporidium parvum (Coccidia, Sporozoa). By using 10--14-day rats as a model, the authors have first provided evidence that even 4-day intestinal cryptosporidiosis may trigger obvious negative changes in the liver and heart, i.e. in the organs where the parasite does not develop. In the infected rats, growth retardation was registered, in addition to liver hypertrophy and partial heart atrophy, and growth retardation. Light and electron microscopies, absorption and fluorescence cytometry, quantitative morphometry, and image analysis were applied. In both hepatocytes and cardiomyocytes, the polyploid cell fraction was seen much increased, with the occurrence of 4c, 8c, and even 16c nuclei. Besides, in the hepatocytes, the amount of glycogen decreased whereas the level of protein increased, along with enhanced nucleolar activity in the nuclei. Unlike, the cardiomyocytes of the infected rats were characterized by protein decrease, in addition to almost two-fold cell body elongation. This is the first documented evidence for serious pathological changes in the extraintestinal organs, caused by the intestinal pathogen C. parvum. Within the first 4 days of infection, both the liver and heart of the host seem to work under stress. It is plausible that on modulating liver and heart ploidy, the intestinal parasitic infection (cryptosporidiosis) may bring about functional impairments of these organs, untypical of early age, leading eventually to long-term consequences in further life of formerly infected individuals.


Evidence for bradykinin potentiation by angiotensin congeners in conscious rats.
It has become increasingly clear that the potent vasoactive peptides bradykinin and angiotensin share a common point of metabolism, i.e., angiotensin-converting enzyme or kininase II, and may interact with prostaglandins to regulate regional blood flow. To establish whether the sensitivity to exogenous bradykinin was affected by the presence of angiotensin, vasodepressor dose-response curves to injected bradykinin were performed in conscious rats before and during a 1-h infusion of angiotensin I (30 ng/min), angiotensin II (30 and 300 mg/min), and [Sar2,Ala8]angiotensin II (5 micrograms/min). All of these induced a parallel leftward shift of the bradykinin dose-response curve of approximately threefold. No similar changes were observed during control infusions of dextrose, similar pressor doses of lysine vasopressin, or norepinephrine. Sensitivity to bradykinin was enhanced by saralasin in normal and nephrectomized rats, suggesting that the antagonist itself was responsible. Similar potentiation was present during both acute (1 h) and chronic infusions (9 days) of angiotensin II and attenuated the effect of a converting-enzyme inhibitor on bradykinin sensitivity. Accordingly, these results suggest a competitive interaction in vivo between angiotensin congeners and bradykinin at a point of bradykinin degradation, probably angiotensin-converting enzyme or kininase II. This is a potential additional mechanism by which these systems may interact to affect regional blood flow and must be considered in the interpretation of results obtained during saralasin infusion.


Twenty-three Cases of Primary Cornea Guttata.
Purpose: An evaluation of the clinical aspects of patients with primary cornea guttata and the morphology of their corneal endothelial cells.Methods: Twenty-three patients who visited Kanazawa Medical University Hospital or related hospitals in Ishikawa or Fukui prefectures and were diagnosed as having primary cornea guttata by slit lamp microscopy and contact specular microscopy underwent an analysis of corneal endothelial cells as well as their clinical status.Results: In 23 cases, the number of men and young people under 30 years of age was low, with 4 and 2 cases respectively. There was no visual impairment due to cornea guttata. In the specular microscopic findings, the size and density of dark areas varied according to each case and there was asymmetry in some cases. The parameters of the endothelial cells, comprising mean cell area, hexagonality, and the coefficient of variation, were almost within normal limits except for one eye of a 79-year-old man who was speculated to be at an early stage of Fuch's endothelial corneal dystrophy. Three cases had received cataract surgery, but showed no statistical changes in endothelial morphology before or after the surgery.Conclusion: Cornea guttata is thought to be a disease which is not uncommon in Japanese people and which does not bring about visual impairment or abnormalities in corneal endothelial cell parameters except for a small number of patients. Such cases with remarkably decreased cell densities should be considered to be at an early stage of Fuch's dystrophy.


Characterization and validation of fluorescent receptor ligands: a case study of the ionotropic serotonin receptor.
The application of fluorescent receptor ligands has become widespread, incited by two important reasons. "Seeing is believing"-it is possible to visualize in real time in live cells ligand-receptor interactions, and to locate the receptors with subcellular precision allowing one to follow, e.g., internalization of the ligand-receptor complex. The high sensitivity of photon detection permits observation of on the one hand receptor-ligand interactions on cells with low, native receptor abundance, and on the other of individual fluorophores unveiling the stochastic properties of single ligand-receptor complexes.The major bottlenecks that impede extensive use of fluorescent ligands are due to possible dramatic changes of the pharmacological properties of a ligand upon chemical modification and fluorophore conjugation, aggravated by the observation that different fluorophores can provoke very dissimilar effects. This makes it virtually impossible to predict beforehand which labelling strategy to use to produce a fluorescent ligand with the desired qualities.Here, we focus on the design, synthesis, and evaluation of a high-affinity fluorescent antagonist for the ionotropic serotonin type-3 receptor.


Variation in personality shaped by evolutionary history, genotype and developmental plasticity in response to feeding modalities in the Arctic charr.
Animal personality has been shown to be influenced by both genetic and environmental factors and shaped by natural selection. Currently, little is known about mechanisms influencing the development of personality traits. This study examines the extent to which personality development is genetically influenced and/or environmentally responsive (plastic). We also investigated the role of evolutionary history, assessing whether personality traits could be canalized along a genetic and ecological divergence gradient. We tested the plastic potential of boldness in juveniles of five Icelandic Arctic charr morphs (Salvelinus alpinus), including two pairs of sympatric morphs, displaying various degrees of genetic and ecological divergence from the ancestral anadromous charr, split between treatments mimicking benthic versus pelagic feeding modalities. We show that differences in mean boldness are mostly affected by genetics. While the benthic treatment led to bolder individuals overall, the environmental effect was rather weak, suggesting that boldness lies under strong genetic influence with reduced plastic potential. Finally, we found hints of differences by morphs in boldness canalization through reduced variance and plasticity, and higher consistency in boldness within morphs. These findings provide new insights on how behavioural development may impact adaptive diversification.


Optimising Two-Stage Vacuum Heat Treatment for a High-Strength Micro-Alloyed Steel in Railway Spring Clip Application: Impact on Microstructure and Mechanical Performance.
The heat treatment process is a vital step for manufacturing high-speed railway spring fasteners. In this study, orthogonal experiments were carried out to obtain reliable optimised heat treatment parameters through a streamlined number of experiments. Results revealed that a better comprehensive mechanical performance could be obtained under the following combination of heat treatment parameters: quenching temperature of 850 °C, holding time of 35 min, medium of 12% polyalkylene glycol (PAG) aqueous solution, tempering temperature of 460 °C, and holding time of 60 min. As one of the most important testing criteria, fatigue performance would be improved with increasing strength. Additionally, a high ratio of martensite to ferrite is proven to improve the fatigue limit more significantly. After this heat treatment process, the metallographic microstructure and mechanical properties satisfy the technical requirements for the high-speed railway practical operation. These findings provide a valuable reference for the practical forming process of spring fasteners.


Structure of full-length wild-type human phenylalanine hydroxylase by small angle X-ray scattering reveals substrate-induced conformational stability.
Human phenylalanine hydroxylase (hPAH) hydroxylates L-phenylalanine (L-Phe) to L-tyrosine, a precursor for neurotransmitter biosynthesis. Phenylketonuria (PKU), caused by mutations in PAH that impair PAH function, leads to neurological impairment when untreated. Understanding the hPAH structural and regulatory properties is essential to outline PKU pathophysiological mechanisms. Each hPAH monomer comprises an N-terminal regulatory, a central catalytic and a C-terminal oligomerisation domain. To maintain physiological L-Phe levels, hPAH employs complex regulatory mechanisms. Resting PAH adopts an auto-inhibited conformation where regulatory domains block access to the active site. L-Phe-mediated allosteric activation induces a repositioning of the regulatory domains. Since a structure of activated wild-type hPAH is lacking, we addressed hPAH L-Phe-mediated conformational changes and report the first solution structure of the allosterically activated state. Our solution structures obtained by small-angle X-ray scattering support a tetramer with distorted P222 symmetry, where catalytic and oligomerisation domains form a core from which regulatory domains protrude, positioning themselves close to the active site entrance in the absence of L-Phe. Binding of L-Phe induces a large movement and dimerisation of regulatory domains, exposing the active site. Activated hPAH is more resistant to proteolytic cleavage and thermal denaturation, suggesting that the association of regulatory domains stabilises hPAH.


Artemisinin mimics calorie restriction to extend yeast lifespan via a dual-phase mode: a conclusion drawn from global transcriptome profiling.
Calorie restriction (CR) promotes longevity among distinct organisms from yeast to mammals. Although CR-prolonged lifespan is believed to associate with enhanced respiratory activity, it is apparently controversial for accelerated energy consumption regardless of insufficient nutrient intake. In reconciling the contradiction of less food supply versus much metabolite dispense, we revealed a CR-based mode of dual-phase responses that encompass a phase of mitochondrial enhancement (ME) and a phase of post-mitochondrial enhancement (PME), which can be distinguished by the expression patterns and activity dynamics of mitochondrial signatures. ME is characterized by global antioxidative activation, and PME is denoted by systemic metabolic modulation. CR-mediated aging-delaying effects are replicated by artesunate, a semi-synthetic derivative of the antimalarial artemisinin that can alkylate heme-containing proteins, suggesting artesunate-heme conjugation functionally resembles nitric oxide-heme interaction. A correlation of artesunate-heme conjugation with cytochrome c oxidase activation has been established from adduct formation and activity alteration. Exogenous hydrogen peroxide also mimics CR to trigger antioxidant responses, affect signaling cascades, and alter respiratory rhythms, implying hydrogen peroxide is engaged in lifespan extension. Conclusively, artesunate mimics CR-triggered nitric oxide and hydrogen peroxide to induce antioxidative networks for scavenging reactive oxygen species and mitigating oxidative stress, thereby directing metabolic conversion from anabolism to catabolism, maintaining essential metabolic functionality, and extending life expectancy in yeast.


Missing defects? A comparison of microscopic and macroscopic approaches to identifying linear enamel hypoplasia.
Linear enamel hypoplasia (LEH), the presence of linear defects of dental enamel formed during periods of growth disruption, is frequently analyzed in physical anthropology as evidence for childhood health in the past. However, a wide variety of methods for identifying and interpreting these defects in archaeological remains exists, preventing easy cross-comparison of results from disparate studies. This article compares a standard approach to identifying LEH using the naked eye to the evidence of growth disruption observed microscopically from the enamel surface. This comparison demonstrates that what is interpreted as evidence of growth disruption microscopically is not uniformly identified with the naked eye, and provides a reference for the level of consistency between the number and timing of defects identified using microscopic versus macroscopic approaches. This is done for different tooth types using a large sample of unworn permanent teeth drawn from several post-medieval London burial assemblages. The resulting schematic diagrams showing where macroscopic methods achieve more or less similar results to microscopic methods are presented here and clearly demonstrate that "naked-eye" methods of identifying growth disruptions do not identify LEH as often as microscopic methods in areas where perikymata are more densely packed.


Therapeutic options for postmenopausal female sexual dysfunction.
Female sexual dysfunction (FSD) is a multidimensional problem combining biological, psychological and interpersonal elements of multiple etiologies. Menopause-related sexual dysfunction may not be reversible without therapy. Hormonal deficiency does not usually decrease in severity over time. Many options are available for the successful treatment of postmenopausal FSD. To review the pharmacological and non-pharmacological therapies available for postmenopausal FSD, focusing on practical recommendations for managing postmenopausal women with sexual complaints, through a literature review of the most relevant publications in this field. PSYCHOSOCIAL THERAPY: This type of therapy (basic counselling, physiotherapy and psychosexual intervention) is considered an adaptable step-by-step approach for diagnostic and therapeutic strategies, normally combined with biomedical interventions to provide optimal outcomes. PHARMACOLOGICAL THERAPY: For postmenopausal FSD, many interventional options are now available, including hormonal therapies such as estrogens, testosterone, combined estrogen/testosterone, tibolone and dehydroepiandrosterone. Menopause and its transition represent significant risk factors for the development of sexual dysfunction. FSD impacts greatly on a patient's quality of life. Consequently, it is receiving more attention thanks to the development of effective treatments. Non-pharmacological approaches should be used first, focusing on lifestyle and psychosexual therapy. If required, proven effective hormonal and non-hormonal therapeutic options are available.


Exhaled nitric oxide among pulpmill workers reporting gassing incidents involving ozone and chlorine dioxide.
The aim of the study was to investigate whether measurement of nitric oxide in exhaled air could be used for assessing the effects of irritants on the respiratory system, in this case recurrent ozone gassing in an occupational setting. The study population comprised bleachery workers (n=56) from a Swedish pulpmill carrying out ozone-based pulp bleaching since 1992 and controls (n=39). Both groups were investigated by measuring NO in exhaled air, methacholine challenge test and answers to a questionnaire concerning history of respiratory symptoms and accidental exposure to ozone peaks. There was no significant difference in NO output between exposed subjects and controls (median 67.2 versus 55.0 nL x min(-1), p=0.64). However, among bleachery workers reporting ozone gassings, the median NO output was 90.0 nL x min(-1) compared to 58.8 nL x min(-1) among those not reporting such incidents (p=0.019). There was no relation between exhaled NO and the prevalence of respiratory symptoms or bronchial hyperresponsiveness. In a multiple regression model, only reported ozone gassings were associated (p=0.016) with NO output. The results indicate an association between previous response to ozone gassing and nitric oxide output. The increased nitric oxide output among the bleachery workers reporting peak ozone exposure may indicate that chronic airway inflammation is present. Further studies are needed to evaluate the extent to which nitric oxide can be used for biological monitoring of respiratory health effects, and to relate it to other markers of airway inflammation.


Hypothalamic neuronal histamine modulates physiological responses induced by interleukin-1 beta.
Dynamic involvement of hypothalamic histamine in ingestive behavior and thermogenesis induced by interleukin-1 beta (IL-1 beta) was examined in rats. Intraperitoneal injection of 0.12 nmol/rat IL-1 beta decreased food and water intake and elevated body temperature. However, depletion of neuronal histamine induced by intraperitoneal injection of 160 mumol/rat alpha-fluoromethylhistidine, a suicide inhibitor of histidine decarboxylase (HDC), attenuated the suppressive effect of IL-1 beta on food intake, facilitated the suppressive effect on drinking, and enhanced the elevating effect on rectal temperature. Intraperitoneal injection of 0.12 nmol/rat IL-1 beta increased hypothalamic histamine turnover rate. The same dose of IL-1 beta also increased activity of HDC and histamine-N-methyltransferase (HMT). These results suggest that IL-1 beta may stimulate synthesis and release of hypothalamic histamine in presynaptic terminals by activation of HDC and facilitate degradation of extracellular histamine by activation of MHT. These changes in the dynamics of hypothalamic histamine modulate IL-1 beta-induced ingestive behavior and body temperature.


Small bowel malignant melanoma presenting as a perforated jejunal diverticulum: a case report and literature review.
Although usually harmless and asymptomatic, jejuno-ileal diverticulae are associated with various non-specific gastrointestinal symptoms, and rarely cause surgical emergencies. This case report describes the presentation and management of a patient with an acute abdomen, whose jejunal diverticulum was perforated. Unexpectedly, histopathological assessment demonstrated malignant melanoma lining the diverticulum. Whether this was primary or metastatic is discussed, together with a synopsis of the literature on small bowel diverticulae.


Prophylactic lymphaticovenous anastomosis (LVA) for preventing lymphedema after sarcoma resection in the lower limb: A report of three cases and literature review.
Patients with soft tissue tumors of the lower extremities are at greater risk to develop postoperative disruption of lymphatic vessels. Currently, there is no widely effective cure for lymphatic dysfunction. Therefore, the best strategy is to prevent it and reconstruct efficient drainage as soon as the original pathway is damaged. We present a report of three prophylactic LVA cases after sarcoma resection in the lower limb, and a literature review to show the feasibility of prophylactic LVAs. The patients were 35, 73, and 77 years old, respectively, at the time of the procedure. All three patients had sarcoma in the medial thigh and underwent radiation therapy before the surgery. The locations of the LVAs include the medial thigh and medial and lateral calf. During the surgery, methylene blue and/or indocyanine green were injected to identify lymphatic vessels. Postoperative recovery was uneventful immediately after the surgery. At follow-up visits, all three patients reported improved functions with no significant swelling in the lower limb. One patient experienced a surgical wound infection that resolved after antibiotic admission. Two patients had a history of cardiac diseases, a major risk factor for developing postoperative lymphedema, but these two patients did not develop lymphedema with the treatment of prophylactic LVAs. These results suggest that prophylactic LVA may be an effective strategy to prevent secondary lymphedema after sarcoma resection. Further investigation is warranted.


[Sterile catheters for newborn infants].
The use of sterile gastroenterologic and umbilical catheters in clinical practice considerably reduces bacterial contamination and postsurgical complications, they may find application in emergency cases as well. Delivery of sterile catheters in sealed packages saves time of medical personnel engaged in obstetric services, ensures long-term storage of products in a sterile form.


DeepDrawing: A Deep Learning Approach to Graph Drawing.
Node-link diagrams are widely used to facilitate network explorations. However, when using a graph drawing technique to visualize networks, users often need to tune different algorithm-specific parameters iteratively by comparing the corresponding drawing results in order to achieve a desired visual effect. This trial and error process is often tedious and time-consuming, especially for non-expert users. Inspired by the powerful data modelling and prediction capabilities of deep learning techniques, we explore the possibility of applying deep learning techniques to graph drawing. Specifically, we propose using a graph-LSTM-based approach to directly map network structures to graph drawings. Given a set of layout examples as the training dataset, we train the proposed graph-LSTM-based model to capture their layout characteristics. Then, the trained model is used to generate graph drawings in a similar style for new networks. We evaluated the proposed approach on two special types of layouts (i.e., grid layouts and star layouts) and two general types of layouts (i.e., ForceAtlas2 and PivotMDS) in both qualitative and quantitative ways. The results provide support for the effectiveness of our approach. We also conducted a time cost assessment on the drawings of small graphs with 20 to 50 nodes. We further report the lessons we learned and discuss the limitations and future work.


Incident atrial fibrillation among Asians, Hispanics, blacks, and whites.
Because the association between atrial fibrillation (AF) and race has only been rigorously compared in population-based studies that dichotomized participants as white or black, it is unclear whether white race confers elevated AF risk or black race affords AF protection. The Healthcare Cost and Utilization Project was used to identify patients receiving hospital-based care in California between January 1, 2005 and December 31, 2009. The association between race and incident AF was examined using Cox proportional hazards models. Interaction analyses were performed to elucidate the mechanism underlying the race-AF association. Among 13 967 949 patients, 375 318 incident AF episodes were observed over a median 3.2 (interquartile range 1.8-4.3) years. In multivariable Cox models adjusting for patient demographics and established AF risk factors, blacks (hazard ratio, 0.84; 95% confidence interval, 0.82-0.85; P<0.001), Hispanics (hazard ratio, 0.78; 95% confidence interval, 0.77-0.79; P<0.001), and Asians (hazard ratio, 0.78; 95% confidence interval, 0.77-0.79; P<0.001) each exhibited a lower AF risk compared with whites. AF risk among whites was disproportionately higher in the absence of acquired cardiovascular risk factors and diminished or reversed in the presence of comorbid diseases. Although Hispanics and Asians also had a lower adjusted risk of incident atrial flutter compared with whites, the risk of flutter was significantly higher among blacks. In a large hospital-based cohort, whites have an increased risk of AF whether compared with blacks, Asians, or Hispanics. The heightened AF risk among whites is most pronounced in the absence of cardiovascular comorbidities.


Glucokinase overexpression restores glucose utilization and storage in cultured hepatocytes from male Zucker diabetic fatty rats.
Zucker diabetic fatty rats develop type 2 diabetes concomitantly with peripheral insulin resistance. Hepatocytes from these rats and their control lean counterparts have been cultured, and a number of key parameters of glucose metabolism have been determined. Glucokinase activity was 4.5-fold lower in hepatocytes from diabetic rats than in hepatocytes from healthy ones. In contrast, hexokinase activity was about 2-fold higher in hepatocytes from diabetic animals than in healthy ones. Glucose-6-phosphatase activity was not significantly different. Despite the altered ratios of glucokinase to hexokinase activity, intracellular glucose 6-phosphate concentrations were similar in the two types of cells when they where incubated with 1-25 mM glucose. However, glycogen levels and glycogen synthase activity ratio were lower in hepatocytes from diabetic animals. Total pyruvate kinase activity and its activity ratio as well as fructose 2,6-bisphosphate concentration and lactate production were also lower in cells from diabetic animals. All of these data indicate that glucose metabolism is clearly impaired in hepatocytes from Zucker diabetic fatty rats. Glucokinase overexpression using adenovirus restored glucose metabolism in diabetic hepatocytes. In glucokinase-overexpressing cells, glucose 6-phosphate levels increased. Moreover, glycogen deposition was greatly enhanced due to the activation of glycogen synthase. Pyruvate kinase was also activated, and fructose-2,6-bisphosphate concentration and lactate production were increased in glucokinase-overexpressing diabetic hepatocytes. Overexpression of hexokinase I did not increase glycogen deposition. In conclusion, hepatocytes from Zucker diabetic fatty rats showed depressed glycogen and glycolytic metabolism, but glucokinase overexpression improved their glucose utilization and storage.


Rectangular tapered short stem excellently preserves proximal bone mineral density preservation than tapered wedge short stem.
Fitmore stem is a rectangular, tapered, short, cementless stem. A characteristic feature of this stem is that it provides rotational stability due to the high medullary occupancy achieved by its rectangular cross-section and thick antero- posterior width. We aimed to investigate the differences in periprosthetic bone remodelling between a rectangular- tapered short stem and a short tapered-wedge stem. Eighty patients who underwent primary total hip arthroplasty using a rectangular-tapered short stem (Fitmore) or a short tapered-wedge stem (Tri-Lock BPS) were enrolled in this study. Bone mineral densities (BMDs) in the seven Gruen zones were evaluated using dual-energy X-ray absorptiometry at baseline, and at 6 and 24 months postoperatively. Peri-prosthetic BMD and clinical factors were assessed and compared. In addition, correlations between periprosthetic BMD changes and stem anteversion error were analyzed using Pearson's correlation coefficient in the two groups. A significantly better postoperative periprosthetic BMD change was found in zones 1 and 7 in the rectangular-tapered group. Additionally, no significant correlation was observed between stem anteversion error and periprosthetic BMD changes in the rectangular-tapered groups. However, in the tapered-wedge group, there were significant negative correlations between the stem anteversion error and BMD changes at 6 months and 24 months in zones 1 and 7. In the rectangular-tapered group, a significantly better postoperative periprosthetic BMD change was found particularly in the region proximal to the stem. Rectangular-tapered short stem can be more resistant to rotation due to higher medullary occupancy and may lead to better periprosthetic BMD than the tapered-wedge short stem, especially in the proximal region of the stem.


Ontogeny of yellow fever 17D vaccine: RNA oligonucleotide fingerprint and monoclonal antibody analyses of vaccines produced world-wide.
Yellow fever 17D vaccines are currently manufactured with approval of the World Health Organization (WHO) in 11 countries. These vaccines have proven highly efficacious and safe. Nevertheless, they have not been fully characterized genetically, a problem for future standardization and modernization of vaccine manufacture now being proposed by WHO. Vaccines in use are derived from two distinct substrains (17D-204 and 17DD) which represent independently maintained passage series from original 17D. In this study, all 17D vaccines produced world-wide were characterized by RNA oligonucleotide fingerprinting. Forty-two large oligonucleotides were compared, and differences from an arbitrarily selected reference strain (produced by Connaught Laboratories in the U.S.A.) were determined. With one exception (vaccine produced in South Africa), fingerprints of vaccines derived from substrain 17D-204 were identical. The South African primary seed differed in position of one oligonucleotide, reflecting a charge shift due to a single base change. This difference occurred within one egg passage; a further change in the South African vaccine occurred within one or two passages from primary seed. No antigenic differences between 17D-204-derived vaccines (including South Africa) were demonstrated by neutralization tests using monoclonal antibody. Vaccines derived from the 17DD substrain consistently differed from 17D-204 vaccines in the absence of one oligonucleotide (No. 37). This change probably occurred during 40 additional egg passages in development of the 17DD vaccines. A clear antigenic difference was shown between 17D-204 and 17DD substrain vaccines using monoclonal antibody. 17DD vaccines showed minor genotypic differences, suggesting a higher degree of genetic instability than 17D-204 vaccines. No oligonucleotide fingerprint differences were found between avian leukosis virus (ALV)-free and ALV-contaminated vaccines. No definite genomic correlate of neurovirulence was defined by fingerprinting strains with a history of encephalitic complications in man or of failure to pass monkey neurovirulence tests. Parent Asibi virus showed several oligonucleotide differences and was serologically distinct from 17D vaccine.


Chemistry of cyclic ADP-ribose and its analogs.
Cyclic ADP-ribose (cADPR), a general mediator involved in Ca2+ signaling, has the characteristic 18-membered ring consisting of an adenine, two riboses and a pyrophosphate, in which the two primary hydroxyl groups of the riboses are linked by a pyrophosphate unit. This review focuses on the chemical synthetic studies of cADPR analogs. These analogs have been used quite effectively in proving the mechanism of cADPR-mediated Ca2+ signaling pathways. These analogs are also expected to be lead structures for the development of drugs. Although cADPR analogs can be synthesized by enzymatic and chemo-enzymatic methods using ADP-ribosyl cyclase, the analogs obtained by these methods are limited due to the substrate-specificity of the enzymes. Consequently, chemical synthetic methods providing a greater variety of cADPR analogs are required. Chemical synthetic studies have demonstrated that the construction of the large 18-membered ring structure is quite difficult. Another problem encountered in the synthesis is the construction of the N1-substituted purine nucleoside structure. The N1-substituted inosine derivatives were prepared by condensation between the N1-(2,4-dinitrophenyl)inosine derivatives and the appropriate amines. For the preparation of the N1-substituted adenosine structures, condensation of the 4-cyano-5-(alkoxymethyleneamino)imidazole nucleosides with the appropriate amines was found to be effective. The first chemical construction of the 18-membered ring was achieved using a bisphosphate-type substrate conformationally restricted in the cyclized product-like syn-form around the N9-glycosyl linkage; however, the yield was inadequate. The key 18-membereding construction was significantly improved by employing the phenylthiophosphate-type substrates. When the substrates were activated by AgNO3 or I2 in the presence of molecular sieves in pyridine, the corresponding 18-membered ring products were obtained in high yields. Using this method as the key step, the chemically and biologically stable cADPR mimic, cADP-carbocyclic-ribose (cADPcR), was synthesized. This method has been applied subsequently to the synthesis of various cADPR analogs.


[Histoplasmosis in the region of the hard palate simulating a lesion caused by Leishmania].
A case of histoplasmosis at the oral cavity simulating mucocutaneous leishmaniasis is reported. The initial suspicion of leishmaniasis was not confirmed due to lack of amastigotes and no reactivity of the Montenegro's skin test. Diagnosis of histoplasmosis was done by Grocott's stained smears and isolation of Histoplasma capsulatum in Sabouraud's-agar slants. Treatment with Amphoterecin B led to complete remission of the lesion.


[Some peculiarities of transfer RNAs hydrolysis by exonuclease A5].
Unusual kinetics of the hydrolysis of the baker's yeast total tRNA and tRNA1Val by exonuclease A5 was found (two stages of the reaction; low initial velocity and still lower velocity of the second reaction stage; high Km values). These peculiarities were shown to be due to the three-dimensional structure of the substrate, which makes a large part of the tRNA molecule resistant to the exonuclease A5 attack. The exhaustive tRNA hydrolysis observed in the presence of very large doses of exonuclease A5 may be explained by a slight (less than 1%) endo-RNAase contamination in the exonuclease A5 preparation. The hydrolysis conditions become optimal, i.e. the exhaustive hydrolysis proceeds most rapidly, when the amount of endo-RNAase admixture reaches 20--30%. The artificial enzyme mixture of such a composition is a convenient reagent for RNA and expecially for tRNA hydrolysis under mild conditions to 5'-mononucleotides. The considerable resistance of tRNA to pure exonuclease A5 action will make it possible porbably to use tRNA instead of circular nucleic acids for endonuclease admixtures estimation in enzyme preparations.


Current advances in research in treatment and recovery: Nicotine addiction.
The health harms of combusted tobacco use are undeniable. With market and regulatory pressures to reduce the harms of nicotine delivery by combustion, the tobacco product landscape has diversified to include smokeless, heated, and electronic nicotine vaping products. Products of tobacco combustion are the main cause of smoking-induced disease, and nicotine addiction sustains tobacco use. An understanding of the biology and clinical features of nicotine addiction and the conditioning of behavior that occurs via stimuli paired with frequent nicotine dosing, as with a smoked cigarette, is important for informing pharmacologic and behavioral treatment targets. We review current advances in research on nicotine addiction treatment and recovery, with a focus on conventional combustible cigarette use. Our review covers evidence-based methods to treat smoking in adults and policy approaches to prevent nicotine product initiation in youth. In closing, we discuss emerging areas of evidence and consider new directions for advancing the field.


Primary malignant tumors of the sphenoidal sinus.
Neoplasms of the paranasal sinuses account for 0.2 to 0.8% of all cancers and for 2 to 3% of head and neck cancers. Primary involvement of the sphenoidal sinus has been reported to occur in only 1 to 2% of all paranasal sinus tumors. Tumors at this site commonly present with cranial neuropathies and require multidisciplinary treatment. During a 21-year period, 27 patients with primary sphenoidal sinus tumors were evaluated. Those with pituitary tumors and fibro-osseous lesions (fibrous dysplasia) were excluded. The records of all remaining patients were reviewed for the following parameters: age, sex, and ethnic origin; presenting symptoms and signs; radiological features and sites of tumor extension; pathological findings; prior treatments; surgical approaches and extent of resection; adjuvant therapy; complications; and patient outcome. A malignant pathological process was found in 26 of the 27 patients. The most common diseases were squamous cell carcinoma (n = 9), adenoid cystic carcinoma (n = 4), chondrosarcoma (n = 3), and neuroendocrine carcinoma (n = 3). Treatments included surgery (n = 1), surgery and radiotherapy (n = 6), surgery and chemotherapy (n = 3), surgery, radiotherapy, and chemotherapy (n = 4), chemotherapy (n = 5), chemotherapy and radiotherapy (n = 3), and radiotherapy (n = 2). The mean follow-up period from initial evaluation was 41 months (range, 2-199 mo). At the last follow-up, 12 patients (48%) were still alive and 13 (52%) had died. The 2-year survival rate for patients with squamous cell carcinoma was 44%. Headache and visual disturbances are common symptoms of sphenoidal sinus tumors. The presence of cranial neuropathies, although common, suggests a less successful outcome. It is advantageous if the surgical team has had experience with both open and endoscopic approaches to the sphenoidal sinus. Patient outcome is maximized by the use of aggressive multidisciplinary therapy.


Distribution of capsular types and antimicrobial susceptibility of invasive isolates of Streptococcus pneumoniae in Colombian children. Pneumococcal Study Group in Colombia.
Streptococcus pneumoniae is the leading bacterial cause of childhood pneumonia in the developing world. This study describes the type distribution and antimicrobial susceptibility of invasive pneumococcal isolates from Colombian children and is part of the Sistema Regional de Vacunas (SIREVA), a PAHO regional initiative designed to determine the ideal serotype composition of a protein polysaccharide pneumococcal conjugate vaccine for use in children less than 5 years old in Latin America. In Colombia, during the study period, centres in Bogota, Medellin, and Cali collected 324 S. pneumoniae isolates from invasive diseases, 238 (73.5%) from children under the age of 2. Pneumonia was the clinical diagnosis in 41.3% cases, meningitis in 41%, and sepsis in 11.2%. The seven most frequent types included 14(21.9%), 5(10.5%), 23F(9.6%), 1(9%), 6B(9%), 19F(7.1%), and 6A(6.2%). The frequency of diminished susceptibility to penicillin (DSP) was 12%, with 8.9% of isolates showing intermediate level resistance and 3.1% showing high level resistance. Among DSP isolates, 23% were also resistant to cefotaxime, 33.3% to erythromycin, 48.7% to chloramphenicol, and 74.3% to trimethoprim/sulfamethoxazole. Multiple resistance was detected in 59% of the isolates that have DSP. Penicillin resistance was associated with types 23F (53.8%) and 14 (25.6%). These data provides information on capsular types prevalent in Colombia that will not only allow the formulation of an ideal vaccine for the region but also reinforce the need for ongoing regional surveillance.


Cholesterol enhances influenza binding avidity by controlling nanoscale receptor clustering.
Influenza virus infects cells by binding to sialylated glycans on the cell surface. While the chemical structure of these glycans determines hemagglutinin-glycan binding affinity, bimolecular affinities are weak, so binding is avidity-dominated and driven by multivalent interactions. Here, we show that membrane spatial organization can control viral binding. Using single-virus fluorescence microscopy, we demonstrate that the sterol composition of the target membrane enhances viral binding avidity in a dose-dependent manner. Binding shows a cooperative dependence on concentration of receptors for influenza virus, as would be expected for a multivalent interaction. Surprisingly, the ability of sterols to promote viral binding is independent of their ability to support liquid-liquid phase separation in model systems. We develop a molecular explanation for this observation via molecular dynamics simulations, where we find that cholesterol promotes small-scale clusters of glycosphingolipid receptors. We propose a model whereby cholesterol orders the monomeric state of glycosphingolipid receptors, reducing the entropic penalty of receptor association and thus favoring multimeric complexes without phase separation. This model explains how cholesterol and other sterols control the spatial organization of membrane receptors for influenza and increase viral binding avidity. A natural consequence of this finding is that local cholesterol concentration in the plasma membrane of cells may alter the binding avidity of influenza virions. Furthermore, our results demonstrate a form of cholesterol-dependent membrane organization that does not involve lipid rafts, suggesting that cholesterol's effect on cell membrane heterogeneity is likely the interplay of several different factors.


Uptake and release of cerium during Fe-oxide formation and transformation in Fe(II) solutions.
Fe-oxides are ubiquitous in soils and sediments and form during Fe(0) corrosion. Depending on redox conditions and solution composition, Fe-oxides such as ferrihydrite, goethite, magnetite, and green rust (GR) may form. These phases typically have high surface area and large affinity for adsorption of trace components. Further, Fe(II)-Fe(III) (hydr)oxides are redox active. Cerium, a member of the lanthanide family, can be used as an analogue for the tri- and tetra-valent actinides found in radioactive waste, expected to be stored in subsurface repositories. In experiments with ferrihydrite, Ce(III) was effectively scavenged from Fe(II)-bearing solutions within 5 min at pH 7. During transformation of ferrihydrite to green rust, however, all Ce(III) was released to solution. By varying initial solution Fe(II):Fe(III) ratio, magnetite and goethite formed together with GR(Na,SO(4)), resulting in decreased Ce(III) release. X-ray photoelectron spectroscopy revealed Ce(III) adsorbed on magnetite. When Fe-oxides were synthesized by air oxidation of Fe(II) solutions at pH 7, GR(Na,SO(4)) played a catalytic role in the oxidation of Ce(III) to Ce(IV) by O(2), removing more than 90% of the dissolved Ce. Transmission electron microscopy revealed that it formed discrete nanocrystals of CeO(2(s)). These results demonstrate that Fe-oxide interaction with radionuclides is likely to depend strongly on the local redox conditions. By analogy with Ce, the trivalent actinides are not expected to be sequestered by preformed GR in anoxic environments. Our results also suggest that trivalent actinides and lanthanides are released when dissimilatory iron reduction of Fe(III)-oxides leads to GR formation However, under oxidizing conditions, GR may influence radionuclide mobility by catalyzing their transformation to a higher oxidation state.


Routine use of microvascular coupling device for arterial anastomosis in breast reconstruction.
Although microvascular coupling devices are used routinely and successfully for venous anastomosis, there are few published reports demonstrating their efficacy for performing arterial anastomosis. It has been the senior author's (C.Y.A.) preference to perform arterial anastomosis using the microvascular coupling device when feasible. All microsurgical breast reconstructions performed by the senior author at the New York University Medical Center between 1998 and 2004 were retrospectively reviewed. A total of 60 patients underwent microsurgical breast reconstruction, of which 20 were bilateral, for a total of 80 flaps. Of the 80 flaps performed, there were 47 muscle-sparing TRAM and 22 deep inferior epigastric perforator (DIEP) flaps, and 11 were superior gluteal flaps. Arterial coupling was successfully performed in 60 of 69 flaps based on the deep inferior epigastric artery (87%) and 2 of 11 gluteal flaps (18%); arterial coupling was performed successfully 62 of 74 times (83.9%) when the thoracodorsal artery was the recipient vessel and never performed when the internal mammary artery was the recipient vessel. The overall flap success rate was 100%. In our large series, we were able to perform a coupled arterial anastomosis in nearly 80% of the cases, without the loss of any flaps. With proper vessel selection and sufficient experience using the microvascular coupler, arterial coupling may be performed in an expeditious, safe, and reliable fashion with minimal morbidity. Though not commonly practiced, use of the coupling device for arterial anastomosis can provide significant time savings, especially in bilateral breast reconstructions.


Advancing Molecular-Guided Surgery through probe development and testing in a moderate cost evaluation pipeline.
Molecular guided oncology surgery has the potential to transform the way decisions about resection are done, and can be critically important in areas such as neurosurgery where the margins of tumor relative to critical normal tissues are not readily apparent from visual or palpable guidance. Yet there are major financial barriers to advancing agents into clinical trials with commercial backing. We observe that development of these agents in the standard biological therapeutic paradigm is not viable, due to the high up front financial investment needed and the limitations in the revenue models of contrast agents for imaging. The hypothesized solution to this problem is to develop small molecular biologicals tagged with an established fluorescent reporter, through the chemical agent approval pathway, targeting a phase 0 trials initially, such that the initial startup phase can be completely funded by a single NIH grant. In this way, fast trials can be completed to de-risk the development pipeline, and advance the idea of fluorescence-guided surgery (FGS) reporters into human testing. As with biological therapies the potential successes of each agent are still moderate, but this process will allow the field to advance in a more stable and productive manner, rather than relying upon isolated molecules developed at high cost and risk. The pathway proposed and tested here uses peptide synthesis of an epidermal growth factor receptor (EGFR)-binding Affibody molecules, uniquely conjugated to IRDye 800CW, developed and tested in academic and industrial laboratories with well-established records for GMP production, fill & finish, toxicity testing, and early phase clinical trials with image guidance.


Azithromycin for the Treatment of Obliterative Bronchiolitis after Hematopoietic Stem Cell Transplantation: A Systematic Review and Meta-Analysis.
Obliterative bronchiolitis (OB) is a major cause of morbidity and mortality in patients undergoing hematopoietic stem cell transplantation (HSCT). Our objective was to perform a systematic review and meta-analysis of the impact of azithromycin on change in forced expiratory volume in 1 second (FEV1). We searched MEDLINE, EMBASE, Web of Science, Cochrane CENTRAL and Scopus databases and included studies that compared azithromycin with placebo or no intervention in the treatment of OB or bronchiolitis obliterans syndrome (BOS) in patients who had undergone allogeneic HSCT. Ninety-one unique publications were identified, and 4 studies met inclusion criteria, with a total of 90 patients. Changes in FEV1 were measured between 12 and 24 weeks after initiation of treatment. The meta-analysis demonstrated a mean increase in FEV1 of 30 mL (95% confidence interval, -260 to +330 mL; P = .82) after initiation of azithromycin. One patient death was reported but not attributed to azithromycin therapy. In conclusion, current evidence can neither support nor refute the use of azithromycin in the treatment of patients who develop OB/BOS after HSCT. Further studies are needed to determine whether azithromycin is beneficial for the treatment of OB/BOS in this setting.


Cytokine gene polymorphisms in Pigeon Breeder's Disease expression.
Exaggerated immunological response to repeated inhalation of organic or chemical dusts may lead to Hypersensitivity Pneumonitis among sensitized individuals. Only a few exposed individuals became ill and disease expression pattern is highly variable which suggest that genetic factors may play a role. To investigate interferon (IFN)-γ, tumour necrosis factor (TNF)-α, interleukin (IL)-6, transforming growth factor (TGF)-ß, and IL-10 gene polymorphisms in a cohort of pigeon breeder's disease (PBD) patients in comparison with exposed but healthy controls and the association with different patterns of disease. We evaluated 40 PBD patients and 70 exposed controls. IFN-γ, TNF-α, IL-6, TGF-ß, and IL-10 polymorphisms were determined by polymerase chain reaction-sequence specific primer amplification. Polymorphism analysis of IFN-γ, TNF-α, IL-6, TGF-ß, and IL-10 genotypes and allele frequencies showed no differences between patients and controls. IFN-γ T/T genotype frequency was increased among patients with chronic presentation (RR=2.33, p=0.047) compared with those with acute/subacute presentation. Also, chronic presenting patients had an increased frequency of IFN-γ T allele (50% vs 22.5%, RR=1.76, p=0.011). No differences were found in TNF-α, IL-6, TGF-ß, and IL-10 genotypes neither allelic frequencies between both groups of patients. IL-6 C/C genotype was more frequent in patients who showed chronic evolution (RR=2.54, p=0.017), when comparing with patients with disease resolution. IFN-γ T/T and the IL-6 C/C genotypes seem to play a role in HP expression due to avian exposure, as their frequencies are increased in chronic presentations or in those with chronic evolution one year after the initial diagnosis, respectively. (Sarcoidosis Vasc Diffuse Lung Dis 2020; 37 (3): e2020004).