17-beta-estradiol induces heat shock proteins in brain arteries and potentiates ischemic heat shock protein induction in glia and neurons.
Estradiol reduces brain injury from many diseases, including stroke and trauma. To investigate the molecular mechanisms of this protection, the effects of 17-beta-estradiol on heat shock protein (HSP) expression were studied in normal male and female rats and in male gerbils after global ischemia. 17-beta-estradiol was given intraperitoneally (46 or 460 ng/kg, or 4.6 microg/kg) and Western blots performed for HSPs. 17-beta-estradiol increased hemeoxygenase-1, HSP25/27, and HSP70 in the brain of male and female rats. Six hours after the administration of 17-beta-estradiol, hemeoxygenase-1 increased 3.9-fold (460 ng/kg) and 5.4-fold (4.6 microg/kg), HSP25/27 increased 2.1-fold (4.6 microg/kg), and Hsp70 increased 2.3-fold (460 ng/kg). Immunocytochemistry showed that hemeoxygenase-1, HSP25/27,and HSP70 induction was localized to cerebral arteries in male rats, possibly in vascular smooth muscle cells. 17-beta-estradiol was injected intraperitoneally 20 minutes before transient occlusion of both carotids in adult gerbils. Six hours after global cerebral ischemia, 17-beta-estradiol (460 ng/kg) increased levels of hemeoxygenase-1 protein 2.4-fold compared with ischemia alone, and HSP25/27 levels increased 1.8-fold compared with ischemia alone. Hemeoxygenase-1 was induced in striatal oligodendrocytes and hippocampal neurons, and HSP25/27 levels increased in striatal astrocytes and hippocampal neurons. Finally, Western blot analysis confirmed that estrogen induced heat shock factor-1, providing a possible mechanism by which estrogen induces HSPs in brain and other tissues. The induction of HSPs may be an important mechanism for estrogen protection against cerebral ischemia and other types of injury.


Anesthesia management with single injection paravertebral block for aorta coarctation in infant.
Thoracotomy causes severe pain in the postoperative period. Perioperative thoracic paravertebral block reduces pain score and may improve outcome after pediatric cardiac surgery. This prospective study was designed for the efficacy and duration of a single level, single injection ultrasound-guided thoracic paravertebral block (TPVB) for fifteen infants undergoing aortic coarctation repair. After approval of the ethical committee and the relatives of the patients, 15 infants who had undergone thoracotomy were enrolled in the study. The patients received 0.5 ml·kg(-1) a bolus 0.25% bupivacaine with epinephrine 1 : 200 000 at T5-6 level after standard general anesthesia induction. Anesthesia depth with Index of Consciousness (IOC) and tissue oxygen saturation with cerebral (rSO2-C) and somatic thoracodorsal (rSO2-S) were monitored. Intraoperative hemodynamic and postoperative hemodynamic and pain scores were evaluated for 24 h after surgery. Face, Legs, Activity, Cry, Consolability (FLACC) score was utilized to measure postoperative pain in the intensive care unit. Rescue 0.05 mg·kg(-1) IV morphine was applied to patients in whom FLACC was >3. The median age of the patients was 4.5 (1-11) months, and the median intraoperative endtidal isoflurane concentration was 0.6% (0.3-0.8). The amount of remifentanil used intraoperatively was 4.5 (2.5-14) μg·kg(-) (1) ·h(-1). Intraoperative heart rate and blood pressure values significantly decreased compared with values detected at 5th, 10th, and 15th min after TPVB application, after incision prior and after cross-clamp (P < 0.01). The median time of first dose of morphine application after block was 320 (185-430) min. The median morphine consumption in 24 h was 0.16 (0.09-0.4) mg·kg(-1). The median length of postoperative intensive care unit and in-hospital stay times was 23 (1-67) h and 4 (1-10) days, respectively. We believe that TPVB, as part of a balanced anesthetic and analgesic regime, provides effective pain relief in patients undergoing aortic coarctation repair.


Familial similarity in body size in an isolated Zapotec-speaking community in the valley of Oaxaca, southern Mexico: estimated genetic and environmental effects.
The study used path analysis to estimate genetic and environmental determinants of familial similarity in anthropometric characteristics among children from a rural subsistence agrarian community in the Valley of Oaxaca in southern Mexico. Anthropometric dimensions included height, weight, arm circumference and triceps skinfold; the BMI was calculated. Parent-offspring pairs (n = 34) were constructed from pedigrees and prior studies using listwise case deletion for parent-male child and parent-female child, respectively. Path coefficients for genetic and environmental effects were computed using linear regression. Age effects were held constant by computing residuals after parameter regression on age and age2. Environmental effects were analysed using a previously published factor analysis of village ecology. By path analysis, the paternal effect was low-to-moderately strong on males (0.21-0.57) but low on females (0.13-0.23). Maternal effects were low, inconsistent and negligible on males (-0.16-0.09), but high on females (0.57-0.81), except for height (0.19). Estimated environmental effects on sibling similarity were low (0.12-0.27). The environment exerts a relatively consistent effect on growth status, and probably accounts for the low path coefficients (i.e. sibling correlations, parent-offspring regressions). Sex-specific effects are apparent in a strong paternal influence on male offspring height and moderate influences on weight, triceps skinfold, and arm circumference. Maternal influences are small on the anthropometric characteristics of males, but are exceptionally large on female weight, triceps skinfold and arm circumference. Estimated environmental effects on sibling similarity are low-to-moderately high.


Effects of PentaLyte and Voluven hemodilution on plasma coagulation kinetics in the rabbit: role of thrombin-fibrinogen and factor XIII-fibrin polymer interactions.
Hydroxyethyl starch (HES) administration has resulted in decreased hemostasis and fibrinogen (FI)-thrombin-(FIIa)-Factor XIII (FXIII) interactions. I proposed to determine the hemostatic effect of hemodilution with PentaLyte (6% HES, mean molecular weight 220 kDa) and Voluven (6% HES, 130 kDa). Rabbits were intravenously administered 20 ml/kg PentaLyte or Voluven (n = 8 per fluid) over 10 min. Plasma was obtained prior to, 1 min and 1 h after hemodilution. Thrombelastography was performed, with clot initiation (R, sec), clot propagation (alpha, degrees), and clot strength (shear elastic modulus, G, dynes/cm2) determined over 20 min. Celite-activated samples had either no additions or addition of FI, FIIa or activated FXIII (FXIIIa) to restore protein content to pre-diluted values. While there were no significant differences between the groups, R significantly decreased 1 h after hemodilution compared with values observed before and 1 min after hemodilution, whereas alpha and G significantly decreased 1 min after hemodilution and then significantly, but only partially, increased 1 h after hemodilution compared with pre-dilution values. Addition of FI, FIIa and FXIIIa significantly decreased R in both groups. alpha and G 1 min after hemodilution were significantly enhanced by FI, FIIa, FXIIIa in both groups; however, 1 h after hemodilution, rabbits administered PentaLyte had alpha and G enhanced only by FI and FXIIIa addition, whereas animals administered Voluven had alpha and G significantly enhanced by FI addition. PentaLyte and Voluven hemodilution initially diminishes FIIa-FI and FXIIIa-fibrin, but within an hour primarily inhibit FXIIIa-fibrin interactions in the rabbit.


Episodes of fissure formation in the Alps: connecting quartz fluid inclusion, fissure monazite age, and fissure orientation data.
Fluid assisted Alpine fissure-vein and cleft formation starts at prograde, peak or retrograde metamorphic conditions of 450-550 °C and 0.3-0.6 GPa and below, commonly at conditions of ductile to brittle rock deformation. Early-formed fissures become overprinted by subsequent deformation, locally leading to a reorientation. Deformation that follows fissure formation initiates a cycle of dissolution, dissolution/reprecipitation or new growth of fissure minerals enclosing fluid inclusions. Although fissures in upper greenschist and amphibolite facies rocks predominantly form under retrograde metamorphic conditions, this work confirms that the carbon dioxide fluid zone correlates with regions of highest grade Alpine metamorphism, suggesting carbon dioxide production by prograde devolatilization reactions and rock-buffering of the fissure-filling fluid. For this reason, fluid composition zones systematically change in metamorphosed and exhumed nappe stacks from diagenetic to amphibolite facies metamorphic rocks from saline fluids dominated by higher hydrocarbons, methane, water and carbon dioxide. Open fissures are in most cases oriented roughly perpendicular to the foliation and lineation of the host rock. The type of fluid constrains the habit of the very frequently crystallizing quartz crystals. Open fissures also form in association with more localized strike-slip faults and are oriented perpendicular to the faults. The combination of fissure orientation, fissure quartz fluid inclusion and fissure monazite-(Ce) (hereafter monazite) Th-Pb ages shows that fissure formation occurred episodically (1) during the Cretaceous (eo-Alpine) deformation cycle in association with exhumation of the Austroalpine Koralpe-Saualpe region (~ 90 Ma) and subsequent extensional movements in association with the formation of the Gosau basins (~ 90-70 Ma), (2) during rapid exhumation of high-pressure overprinted Briançonnais and Piemontais units (36-30 Ma), (3) during unroofing of the Tauern and Lepontine metamorphic domes, during emplacement and reverse faulting of the external Massifs (25-12 Ma; except Argentera) and due to local dextral strike-slip faulting in association with the opening of the Ligurian sea, and (4) during the development of a young, widespread network of ductile to brittle strike-slip faults (12-5 Ma). The online version contains supplementary material available at 10.1186/s00015-021-00391-9.


Electromyographic study of the upper, middle and lower portion of the trapezius muscle in the circumduction movement of the arm on a shoulder wheel apparatus.
The upper, middle and lower portions of the trapezius muscle were analysed electromyographically (using a TECA TE-4 electromyograph and a single coaxial needle) in 20 healthy volunteers, during a circumduction movement of the arm, on a shoulder wheel apparatus. All three portions of the muscle were active, but the elevation phase of the arm was more pronounced in movements performed clockwise while the lowering phase was more manifest in movements performed counterclockwise. The position of the wheel gauntlet did not interfere significantly with the results.


Cotton straw biochar and Bacillus compound biofertilizer decreased Cd migration in alkaline soil: Insights from relationship between soil key metabolites and key bacteria.
Cadmium (Cd) contamination greatly impacts soil health and ecological environment. In recent years, cotton straw biochar and Bacillus compound biofertilizer have been paid much attention in the remediation of Cd-contaminated soils. In this study, the effects of cotton straw biochar (3%, w/w) and Bacillus compound biofertilizer (1.5%, w/w) on the Cd fractions, Cd migration, bacterial community succession, and metabolites in the soils with different concentrations of Cd (1, 2, and 4 mg kg-1) were explored. The results showed that the relative abundance of Actinobacteriota, Acidobacteriota, Firmicutes, and Cyanobacteric and soil enzyme activities in Cd-contaminated soils decreased, and the soil metabolic pathways also changed compared with those in the control. After the application of cotton straw biochar and Bacillus compound biofertilizer, the soil available Cd concentration in Cd-contaminated soils decreased, and many exchangeable and carbonate-bound Cd were transformed into residual Cd, which decreased the bioavailability of Cd in the soil and the accumulation of Cd in cotton organs. In addition, the application of cotton straw biochar and Bacillus compound biofertilizer improved the activity of soil enzymes and the abundance of dominant bacteria and stimulated Verrucomicrobiota, Methylomirabilota, and Cyanobacteria to secrete organic acids and amino acid compounds, which decreased the toxicity of Cd. Besides, compared with cotton straw biochar, Bacillus compound biofertilizer was more effective in immobilizing Cd and improving soil environment. This study provides guidance for the remediation of Cd-contaminated alkaline soil, and makes contributions to the soil health and sustainable development.


Induction of alopecia areata in C3H/HeJ mice using polyinosinic-polycytidylic acid (poly[I:C]) and interferon-gamma.
Alopecia areata (AA) is a chronic, relapsing hair-loss disorder that is considered to be a T-cell-mediated autoimmune disease. Several animal models for AA have been created to investigate the pathophysiology and screen for effective therapeutic targets. As C3H/HeJ mice develop AA spontaneously in a low frequency, a novel animal model is needed to establish an AA-like condition faster and more conveniently. In this study, we present a novel non-invasive AA rodent model that avoids skin or lymph-node cell transfer. We simply injected C3H/HeJ mice subcutaneously with interferon-gamma (IFNγ) along with polyinosinic:polycytidylic acid (poly[I:C]), a synthetic dsRNA, to initiate innate immunity via inflammasome activation. Approximately 80% of the IFNγ and poly(I:C) co-injected mice showed patchy AA lesions after 8 weeks. None of the mice displayed hair loss in the IFNγ or poly(I:C) solely injection group. Immunohistochemical staining of the AA lesions revealed increased infiltration of CD4+ and CD8+ cells infiltration around the hair follicles. IFNγ and poly(I:C) increased the expression of NLRP3, IL-1β, CXCL9, CXCL10, and CXCL11 in mouse skin. Taken together, these findings indicate a shorter and more convenient means of AA animal model induction and demonstrate that inflammasome-activated innate immunity is important in AA pathogenesis.


Amyloid-β-Induced Transglutaminase 2 Expression and Activities are Modulated by 2-Pentadecyl-2-Oxazoline in Mouse and Human Microglial Cell Lines.
Transglutaminase 2 is an ubiquitously multifunctional enzyme and the most widely studied of the transglutaminase family. Consistent with its role in promoting post-translational modifications of proteins, Transglutaminase 2 is involved in many physiological processes such as apoptosis, signal transduction, and cellular adhesion. Several findings indicate that Transglutaminase 2 plays a role in the pathological processes of various inflammation-related diseases, including neurodegenerative diseases. We tested the potential modulatory effects on amyloid-β-induced Transglutaminase 2 expression and activities of 2-pentadecyl-2-oxazoline, a plant-derived agent, which has shown effectiveness against chronic pain and associated neuropsychiatric disorders, both in mouse and human microglial cell lines. We used biochemistry, molecular and cell biology techniques to evaluate the potential modulatory effects on amyloid-β-induced Transglutaminase 2 expression and activities of 2- pentadecyl-2-oxazoline in mouse and human microglial cell lines. 2-pentadecyl-2-oxazoline was able to modulate amyloid-β-induced Transglutaminase 2 expression and activities in mouse and human microglial cell lines. Transglutaminase 2 confirms its role as a neuroinflammation marker, the inhibition of which could be a potential preventive and therapeutic approach, while 2-pentadecyl-2-oxazoline is a potent modulator of the amyloid-β-induced Transglutaminase 2 expression and activities in mouse and human microglial cell lines.


Nonspecific inhibitors of coronavirus OC43 haemagglutination in human sera.
Antibodies against the human coronavirus OC43 in human sera were measured by haemagglutination inhibition (HI), complement fixation (CF), and radial diffusion-haemolysis in gel (HIG) techniques. The apparent HI titres in a fraction of sera with no antibodies detectable by the two other methods were found to be reduced considerably after treating the sera with phospholipase C (PLC). The PLC treatment also reduced the apparent HI titres in some sera containing variable amounts of CF and/or HIG antibodies, but did not affect antibody determinations by the two latter methods. These results suggest that false positive results can be obtained in the OC43 HI test unless the sera are treated with phospholipase C before the assay.


The Patient Acceptable Symptomatic State in Primary Anterior Cruciate Ligament Reconstruction: Predictors of Achievement.
To identify thresholds for patient acceptable symptomatic state (PASS) achievement in a cohort of primary anterior cruciate ligament reconstruction (ACLR) recipients, and to identify factors predictive of PASS achievement. A prospective clinical registry was queried for primary ACLR patients from January 2014 to April 2017 with serial patient-reported outcome measure (PROM) completion at 6, 12, and 24 months. Exclusion criteria included significant concomitant procedures. Knee-based PROMs included the International Knee Documentation Committee (IKDC) score and Knee Injury and Osteoarthritis Outcome Score (KOOS) subscores. PASS threshold values were calculated using receiver operating characteristic (ROC) curves with area under the curve (AUC) analysis. A stepwise multivariate regression identified preoperative and operative predictors of PASS achievement. A total of 144 primary ACLR patients (30.86 ± 12.78 years, body mass index 25.51 ± 4.64, 41.0% male) were included in the analysis. PASS threshold values were established using ROC curve analysis, all of which exceeded 0.7 on AUC analysis (0.742 to 0.911). Factors impacting odds of PASS achievement in the ACLR cohort included preoperative exercises (odds ratio [OR] 2.95 to 4.74, P = .003 to .038), worker's compensation status (OR 0.25 to 0.28, P = .014 to .033), preoperative scores (OR 1.03 to 1.07, P = .005 to <.001), iliotibial band tenodesis (OR 11.08, P = .010), and anteromedial approach (OR 18.03 to 37.05, P < .001). Factors predictive of PASS achievement in recipients of primary ACLR include functional status (e.g., preoperative exercise, preoperative KOOS Sport/Recreation score), worker's compensation status, technique (e.g., anteromedial) and preoperative PROMs. The results of our study are important in better informing shared decision-making models and improving evidence-based guidelines to optimize patient outcomes.


Evaluation of chiral discrimination of highly negatively charged enantiomers by quaternary ammonium beta-cyclodextrin using 1H NMR.
The positively charged quaternary ammonium cyclodextrin, QA-beta-CD, was previously used as a chiral selector to achieve baseline resolution of two of the dianionic enantiomers of disodium 3-(p-isothiocyanatophenoxy)-3-(p-isothiocyanatophenyl)propane-1,2-disulfate by capillary electrophoresis. The basis of the chiral discrimination between QA-beta-CD and the enantiomers was investigated by (1)H NMR spectroscopy. COSY and NOESY spectra were used to infer the role that molecular interactions and the stereocenters have upon association of QA-beta-CD with the enantiomers. A parallel two-step complexation model is used to rationalize the NMR and the chiral discrimination observed during separation of the enantiomers.


A Clinicopathological and Prognostic Analysis of PD-L2 Expression in Surgically Resected Primary Lung Squamous Cell Carcinoma.
Immunotherapy targeting programmed cell death-1 (PD-1) and programmed death-ligand 1 (PD-L1) has shown dramatic therapeutic effects for lung squamous cell carcinoma (SCC), and PD-L1 expression has been shown not only to be a predictive biomarker for response to immunotherapy but also a prognostic factor for lung SCC. However, the clinical significance of programmed death-ligand 2 (PD-L2), another PD-1 ligand, remains unclear. Therefore, we analyzed PD-L2 expression by immunohistochemistry in surgically resected primary lung SCC. PD-L1 and PD-L2 expression on tumor cells were analyzed in 211 primary lung SCC specimens by immunohistochemistry. Additionally, numbers of CD3+, CD4+, and CD8+ tumor-infiltrating lymphocytes were also examined. The rates of positive PD-L2 expression were 77.3% and 67.3% using 5% and 10% cut-off values, respectively. Low PD-L2 expression on tumor cells was statistically associated with histological type (non-keratinizing/keratinizing) and lymphatic invasion. PD-L2-positive patients had significantly longer postoperative survival time (log-rank test; p = 0.0170 at 5% cut-off and p = 0.0500 at 10% cut-off). Furthermore, survival analysis according to PD-L1 and PD-L2 expression revealed that PD-L1-positive and PD-L2-negative patients had the most unfavorable prognosis. PD-L2 protein expression was associated with prognosis in primary lung SCC patients. PD-L2 expression might be a potential biomarker for response to PD-1/PD-L1-targeted immunotherapy, which should be investigated in future studies.


A neural mass model to predict electrical stimulation evoked responses in human and non-human primate brain.
Deep brain stimulation (DBS) is a valuable tool for ameliorating drug resistant pathologies such as movement disorders and epilepsy. DBS is also being considered for complex neuro-psychiatric disorders, which are characterized by high variability in symptoms and slow responses that hinder DBS setting optimization. The objective of this work was to develop an in silico platform to examine the effects of electrical stimulation in regions neighboring a stimulated brain region. We used the Jansen-Rit neural mass model of single and coupled nodes to simulate the response to a train of electrical current pulses at different frequencies (10-160 Hz) of the local field potential recorded in the amygdala and cortical structures in human subjects and a non-human primate. We found that using a single node model, the evoked responses could be accurately modeled following a narrow range of stimulation frequencies. Including a second coupled node increased the range of stimulation frequencies whose evoked responses could be efficiently modeled. Furthermore, in a chronic recording from a non-human primate, features of the in vivo evoked response remained consistent for several weeks, suggesting that model re-parameterization for chronic stimulation protocols would be infrequent. Using a model of neural population activity, we reproduced the evoked response to cortical and subcortical stimulation in human and non-human primate. This modeling framework provides an environment to explore, safely and rapidly, a wide range of stimulation settings not possible in human brain stimulation studies. The model can be trained on a limited dataset of stimulation responses to develop an optimal stimulation strategy for an individual patient.


Comparison of gas clearance and radioactive microspheres for pancreatic blood flow measurement.
Measurement of pancreatic blood flow (PBF) is technically demanding. Although radiolabeled microspheres are considered the "gold standard" for PBF assessment, they have practical limitations. In the current study, H2 and xenon-133 gas clearance techniques were adapted to PBF measurement and compared to radiolabeled microsphere techniques. Simultaneous measurements of PBF were made using either hydrogen or xenon gas washout and radiolabeled microspheres. Measurements were made under basal, vasoconstricted (vasopressin 2U i.v. or nicotine 4 micrograms/kg/h) and stimulated (secretin 125 ng/kg/h or 2 U/kg i.v.) conditions (random order). Mean PBF was 26.9 +/- 5.3, 50.5 +/- 2.3 and 27.6 +/- 5.2 ml/min/100 g basally, 36.9 +/- 8.0, 90.1 +/- 18.9, and 81.7 +/- 14.5 ml/min/100 g in the stimulated state, and 24.2 +/- 7.8, 25.0 +/- 3.5, and 14.9 +/- 7.5 ml/min/100 g in the vasoconstricted state for hydrogen gas clearance, xenon gas clearance, and radiolabeled microspheres, respectively. The H2 clearance technique resulted in tissue trauma, was complicated by frequent electrode displacement, and correlated poorly (r2 = 0.36, p greater than 0.05) with microsphere values. In contrast, xenon clearance measurement had no apparent effect on the pancreas and correlated well (r2 = 0.83, p less than 0.01) with microsphere data. We conclude that xenon clearance offers an attractive, validated alternative to radiolabeled microspheres for measuring pancreatic blood flow.


Time and experience are independent determinants of representational drift in CA1.
In this issue of Neuron, Khatib et al.1 and Geva et al.2 present complementary and breakthrough discoveries demonstrating that elapsed time and active experience independently affect unique aspects of representational drift in the hippocampus.


Antiretroviral drug treatment interruption in human immunodeficiency virus-infected adults: Clinical and pathogenetic implications for the central nervous system.
Interruption of antiretroviral treatment provides a well-defined 'experimental' paradigm to study the dynamics of central nervous system (CNS) infection and host responses in relation to those of systemic infection. We review our experience with 12 subjects (9 who were viremic and three with suppressed infection at baseline) followed longitudinally with serial lumbar punctures and neurological evaluations after stopping their antiretroviral treatments. All but two subjects exhibited an increase in cerebrospinal fluid (CSF) HIV RNA. Approximately half of the cohort developed a substantial, though asymptomatic, CSF lymphocytic pleocytosis with CSF counts rising to 30-60 cells/microL in five of the subjects. Subjects with higher CSF cell counts exhibited higher CSF HIV concentrations. We interpret the relationship of CSF HIV concentrations and pleocytosis in the context of a simple model of virus and cell exchange between blood and CSF. The proportionally greater increase in CSF HIV after treatment interruption indicates that CSF HIV infection is often more effectively suppressed by combination antiretroviral therapy than is systemic infection.


Myelodysplastic syndromes.
Myelodysplastic syndromes (MDS) are a family of myeloid cancers with diverse genotypes and phenotypes characterized by ineffective haematopoiesis and risk of transformation to acute myeloid leukaemia (AML). Some epidemiological data indicate that MDS incidence is increasing in resource-rich regions but this is controversial. Most MDS cases are caused by randomly acquired somatic mutations. In some patients, the phenotype and/or genotype of MDS overlaps with that of bone marrow failure disorders such as aplastic anaemia, paroxysmal nocturnal haemoglobinuria (PNH) and AML. Prognostic systems, such as the revised International Prognostic Scoring System (IPSS-R), provide reasonably accurate predictions of survival at the population level. Therapeutic goals in individuals with lower-risk MDS include improving quality of life and minimizing erythrocyte and platelet transfusions. Therapeutic goals in people with higher-risk MDS include decreasing the risk of AML transformation and prolonging survival. Haematopoietic cell transplantation (HCT) can cure MDS, yet fewer than 10% of affected individuals receive this treatment. However, how, when and in which patients with HCT for MDS should be performed remains controversial, with some studies suggesting HCT is preferred in some individuals with higher-risk MDS. Advances in the understanding of MDS biology offer the prospect of new therapeutic approaches.


One-Step Biallelic and Scarless Correction of a β-Thalassemia Mutation in Patient-Specific iPSCs without Drug Selection.
Monogenic disorders (MGDs), which are caused by single gene mutations, have a serious effect on human health. Among these, β-thalassemia (β-thal) represents one of the most common hereditary hematological diseases caused by mutations in the human hemoglobin β (HBB) gene. The technologies of induced pluripotent stem cells (iPSCs) and genetic correction provide insights into the treatments for MGDs, including β-thal. However, traditional approaches for correcting mutations have a low efficiency and leave a residual footprint, which leads to some safety concerns in clinical applications. As a proof of concept, we utilized single-strand oligodeoxynucleotides (ssODNs), high-fidelity CRISPR/Cas9 nuclease, and small molecules to achieve a seamless correction of the β-41/42 (TCTT) deletion mutation in β thalassemia patient-specific iPSCs with remarkable efficiency. Additionally, off-target analysis and whole-exome sequencing results revealed that corrected cells exhibited a minimal mutational load and no off-target mutagenesis. When differentiated into hematopoietic progenitor cells (HPCs) and then further to erythroblasts, the genetically corrected cells expressed normal β-globin transcripts. Our studies provide the most efficient and safe approach for the genetic correction of the β-41/42 (TCTT) deletion in iPSCs for further potential cell therapy of β-thal, which represents a potential therapeutic avenue for the gene correction of MGD-associated mutants in patient-specific iPSCs.


Quantifiable analysis of cellular pathway inhibition of a Nedd8-activating enzyme inhibitor, MLN4924, using AlphaScreen.
Cellular effects of a Nedd8-activating enzyme (NAE) inhibitor, MLN4924, using the AlphaScreen format were explored. MLN4924 acts as a substrate-assisted inhibitor of NAE by forming a tight binding Nedd8-MLN4924 adduct. The inhibited enzyme can no longer transfer Nedd8 downstream to modify and activate the E3 cullin-RING ligases. This results in the stabilization of proteins regulated by the proteasome, leading to cell death. These studies monitored the endogenous cellular changes to NAE∼Nedd8 thioester, the formation of the Nedd8-MLN4924 adduct, and the reduction in the Cul1-Nedd8. Lysates derived from MLN4924-treated HCT116 cells showed that whereas the β-subunit of NAE remained constant, reductions of both NAE∼Nedd8 thioester and Cul1-Nedd8 levels occurred with a concomitant rise of the adduct. Moreover, the formation of the Nedd8-MLN4924 adduct was approximately stoichiometric with the concentration of NAEβ. Higher density 384-well cell-based assays illustrated the kinetics of enzyme inactivation across a wider range of MLN4924 concentrations, showing a rapid loss of NAE∼Nedd8 thioester and Cul1-Nedd8. The reduction of NAE∼Nedd8 thioester precedes the loss of Cul1-Nedd8 at twice the rate. Finally, these results clearly demonstrate the utility of the homogeneous assay for quantitative assessment of these endogenous cellular components in a 384-well plate in response to inhibition of NAE by MLN4924.


Reasons for noneffectiveness of organ donation programs in five hospitals in the state of Espírito Santo, Brazil.
Although, in absolute numbers, more organ transplantations are performed in Brazil than in many other countries, the donation rate (the number of donors per million individuals) remains quite low. The aim of this study was to determine the reasons for nonfulfillment of organ donations in five hospitals in the State of Espirito Santo, Brazil. This knowledge could help to identify strategies and implement changes to improve the organ donation process and to increase the rate in Brazil and other countries that face similar challenges. This prospective study collected data including the numbers of patients with suspected brain death (both confirmed and nonconfirmed), effective, and of nonfulfilled donations. Data were collected using forms completed daily by the Intra-Hospital Commission for Organ and Tissue Donation of the hospitals during the period from March to August 2010. There were 1386 deaths. The protocol for brain death diagnosis was initiated in 2.8% of them, but only 28.21% of confirmed brain death patients became effective donors. The causes of nondonation differed among the hospitals surveyed, suggesting that different strategies should be implemented to combat the specific weaknesses of each service, which must be continuously evaluated to increase the efficiency of the organ donation process.


Synthesis and initial evaluation of quinoline-based inhibitors of the SH2-containing inositol 5'-phosphatase (SHIP).
Recently, inhibition of the SH2-containing inositol 5'-phosphatase 1 (SHIP1) has become an attractive strategy for facilitating engraftment of MHC-I mismatched bone marrow grafts, increasing the number of adult stem cells in vivo, and inducing mobilization of hematopoietic stem cells. Utilizing high-throughput screening, two quinoline small molecules (NSC13480 and NSC305787) that inhibit SHIP1 enzymatic activity were discovered. New syntheses of these inhibitors have been developed which verified the relative stereochemistry of these structures. Utilizing this synthetic route, some analogs of these quinolines have been prepared and tested for their ability to inhibit SHIP. These structure activity studies determined that an amine tethered to the quinoline core is required for SHIP inhibition. SHIP inhibition may explain the antitumor effects of similar quinoline amino alcohols and provides an impetus for further synthetic studies in this class of compounds.


A rare case of arterial thrombosis in a 37-year-old male with Factor V Leiden mutation.
The classic triad of symptoms seen in chronic mesenteric ischaemia is post-prandial pain, sitophobia (fear of food) and progressive weight loss. Patients with mesenteric ischaemia secondary to a prothrombotic state such as that rendered by the Factor V Leiden mutation, are substantially younger than the typical elderly patient in whom embolic disease triggered by atrial fibrillation is the main underlying cause. This is one such case report documenting arterial thrombosis in a 37-year-old male with a subsequently identified heterozygous Factor V Leiden mutation. Factor V Leiden mutation is a contributing risk factor in cases of small bowel infarction.


Genetic variations in idiopathic pulmonary fibrosis and patient response to pirfenidone.
Genetic variations in Idiopathic Pulmonary Fibrosis (IPF) affect survival and outcomes. Current antifibrotic agents are managed based on the patient's reported side effects, although certain single nucleotide polymorphisms (SNPs) might alter treatment response and survival depending on the antifibrotic administered. This study investigated variations in response and outcomes to pirfenidone based on patients-specific genetic profiles. Retrospective clinical data were collected from 56 IPF patients and had blood drawn for DNA extraction between 7/2013 and 3/2016, with the last patient followed until 10/2018. Nine SNPs were selected for pharmacogenetic investigation based on prior associations with IPF treatment outcomes or implications for pirfenidone metabolism. Genetic variants were examined in relation to clinical data and treatment outcomes. Of the 56 patients, 38 were males (67.85%). The average age of IPF at diagnosis was 66.88 years. At the initiation of pirfenidone, the average percent predicted FVC was 70.7%, and the average DLCO percent predicted was 50.02% (IQR 40-61%). Among the genetic variants tested, the TOLLIP rs5743890 risk allele was significantly associated with improved survival, with increasing pirfenidone duration. This finding was observed with CC or CT genotype carriers but not for those with the TT genotype (p = 0.0457). Similarly, the TGF-B1 rs1800470 risk allele was also significantly associated with improved survival with longer pirfenidone therapy (p = 0.0395), even though it was associated with disease progression. This pilot study suggests that in IPF patients, the TOLLIP rs5743890 genotypes CC and CT, as well as TGF-B1 rs 1800470 may be associated with increased survival when treated with pirfenidone.


Oral Corticosteroid Therapy as an Adjuvant Treatment for Acute Bleeding in Hemophilia Patients With High Titer Inhibitors.
Treatment options exist for patients with severe hemophilia and high titer factor VIII inhibitors but is often inadequate. Few studies have been conducted to evaluate the utility of short-term corticosteroid therapy for improvement in bleeding complications and temporary or sustained resolution of inhibitors in these patients. We describe 2 patients with acute muscular hematomas successfully treated with 4 to 5 days of oral adjuvant corticosteroid therapy resulting in improvement in their acute bleeds and temporary reduction of inhibitors. Thus, the addition of corticosteroids to traditional therapy of hemophilia with inhibitors may be beneficial in some patients. In those with impending compartment syndrome steroids may improve edema and bleeding symptoms preventing the need for surgical interventions.


Isolation, characterization and comparison of mammalian epidermal prekeratins.
1. The lower living layers of mammalian epidermis contain a cytoplasmic tonofilament protein, prekeratin, believed to be the precursor of the keratin which is found in the outer dead cell layer or stratum corneum. 2. Prekeratin is distinguished by its property of being extractable from epidermis homogenized in the presence of citric acid trisodium citrate buffer pH 2.65. 3. In the present study we have compared the epidermal prekeratins from ten mammalian species and have shown them to be of similar amino acid composition. 4. Conditions have been established for studying the immunology of these insoluble proteins and examination of their immunological properties has shown that they are similar to one another but that their antigenic determinants are different from those of callus keratin. 5. The SDS polyacrylamide gel electrophoretic patterns of these proteins differ widely and we have also demonstrated anatomical site variation by this method.


Leiomyosarcoma: a rare presentation as multifocal lesion.
Leiomyosarcoma is a rare type of connective tissue cancer, accounting for 5-10% of all soft tissue sarcomas. We present a case of leiomyosarcoma as unusual multifocal presentation. Retroperitoneal, mediastinal, pulmonary, uterine and bony regions were all involved at the time of presentation. The liver was normal without detected lesions.


Regional differences in behaviors of fascicle and tendinous tissue of the biceps femoris long head during hamstring exercises.
The biceps femoris long head (BFLH) gains its properties from internal elements (fascicles and tendinous tissues) which behaviors remain poorly understood across BFLH regions and dynamic tasks. The aim of this study was to assess the in vivo behaviors of fascicles and tendinous tissue in the proximal and distal regions of BFLH during different dynamic knee and hip tasks. Twenty males performed the Nordic hamstring exercise (NHE) (n = 9) and Romanian deadlift (RDL) (n = 11). Activation of the BFLH was assessed using surface electromyography signals. Ultrasound images of BFLH and kinematic data were used to estimate the interaction between fascicle and tendinous tissue. The fascicles changed less in length (p < 0.05) and contributed less to BFLH length change (p < 0.05) in NHE with higher activation (p < 0.05) relative to RDL. The higher pennation angle changes of BFLH were found in distal region compared to proximal region in both tasks (p < 0.05), while the activation of distal region was higher than activation of proximal region in NHE (p < 0.05). The BFLH length change was primarily contributed by the tendinous tissue during dynamic resistance tasks, and was contributed less by fascicles which operated more isometrically in knee-dominant NHE with higher activation relative to hip-dominant RDL. Regional differences in pennation angle change and activation during dynamic tasks suggest potential regional differences in the mechanical function of BFLH, warranting further investigation.


Preparation of Cytolysin A (ClyA) Nanopores.
The ionic currents passing through nanopores can be used to sequence DNA and identify molecules at the single-molecule level. Recently, researchers have started using nanopores for the detection and analysis of proteins, providing a new platform for single-molecule enzymology studies and more efficient biomolecular sensing applications. For this approach, the homo-oligomeric Cytolysin A (ClyA) nanopore has been demonstrated as a powerful tool. Here, we describe a simple protocol allowing the production of ClyA nanopores. Monomers of ClyA are expressed in Escherichia coli and oligomerized in the presence of detergent. Subsequently, different oligomer variants are electrophoretically resolved and stored in a gel matrix for long-term use.


The relationship between circulating natural killer cells after reduced intensity conditioning hematopoietic stem cell transplantation and relapse-free survival and graft-versus-host disease.
Natural killer cells are known to have anti-tumor activity in haploidentical hematopoietic stem cell transplantation. We hypothesized that reconstituted circulating natural killer cells may be associated with improved relapse-free survival after HLA-matched hematopoietic stem cell transplantation. Serial peripheral blood absolute natural killer cell counts were prospectively measured by flow cytometry of lymphocytes expressing CD56 and CD16 in 167 patients. Cluster analysis was used at engraftment and 60 days post-transplant to distinguish patients with high and low absolute natural killer cell counts. At engraftment 80 patients had high counts (> 22.2/mm3) and 43 had low counts. At 60 days post-transplant 84 patients had high counts (> 18.2/mm3) and 38 had low counts. The primary study end-points were death, relapse and acute graft-versus-host disease. The median follow-up was 373 days (range, 67-1767). Among patients given reduced intensity conditioning, a low absolute natural killer cell count at 60 days post-transplant was independently associated with relapse [adjusted hazard ratio (AHR) = 28.4, 95% confidence interval (CI) 4.3-186.4] and death (AHR = 17.5, 95% CI 4.3-71.3). Furthermore, patients given reduced intensity conditioning who had a high absolute natural killer cell count at 60 days had a significantly better 1-year survival than those with a low count by Kaplan-Meier analysis (83% vs. 11%, p<0.001). Multivariate analysis confirmed that low 60-day absolute natural killer count in patients given reduced intensity conditioning was independently associated with an increase in relapse or death (AHR = 20.22, 95% CI 4.76-85.40). In contrast, there was no significant association between 60-day absolute natural killer cell counts and clinical outcomes in patients receiving myeloablative conditioning. There was no significant association between absolute natural killer cell count and graft-versus-host disease. High natural killer cell reconstitution is associated with reduced relapse and death without an increased incidence of graft-versus-host-disease after reduced intensity conditioning allogeneic hematopoietic stem cell transplantation. Measuring reconstituted natural killer cells expressing CD56(+)/CD16(+) post-transplant may have novel prognostic and therapeutic implications.


Correction: Single-cell genomics shedding light on marine Thaumarchaeota diversification.
An amendment to this paper has been published and can be accessed via a link at the top of the paper.


Modified Selective Neurectomy for the Treatment of Post-Facial Paralysis Synkinesis.
To address functional and smile dysfunction associated with post-facial paralysis synkinesis, the senior author (B.A.) has offered "modified selective neurectomy" of the lower division of the facial nerve as a long-term solution. This article examines technical considerations and outcomes of this procedure. A retrospective review was conducted of patients who underwent modified selective neurectomy of buccal and cervical branches of the facial nerve performed by a single surgeon over a 4½-year period. House-Brackmann facial grading scores, electronic clinician-graded facial function scale, and onabotulinumtoxinA (botulinum toxin type A) dosages were examined before and after the procedure. Sixty-three patients underwent modified selective neurectomy between June 20, 2013, and August 12, 2017. There were no serious complications. The revision rate was 17 percent. Temporary oral incompetence was reported in seven patients (11 percent) postoperatively. A statistically significant improvement was achieved in electronic clinician-graded facial function scale analysis of nasolabial fold depth at rest, oral commissure movement with smile, nasolabial fold orientation with smile, nasolabial depth with smile, depressor labii inferioris lower lip movement, midfacial synkinesis, mentalis synkinesis, platysmal synkinesis, static score, dynamic score, synkinesis score, periocular score, lower face and neck score, and midface and smile score. There was a significant decrease in botulinum toxin type A dosage and House-Brackmann score after surgery. Modified selective neurectomy of the buccal and cervical divisions of the facial nerve is an effective long-term treatment for smile dysfunction in patients with post-facial paralysis synkinesis. Therapeutic, IV.


The role of standard technique in modern sinus surgery.
This article reviews the most commonly performed intranasal and extranasal sinus operations. The indications, complications, and functional results of each are presented. Endoscopic approaches should be used in conjunction with standard techniques in order to supplement and improve the efficiency of standard techniques of sinus surgery.


Chromosome 2p14 is linked to susceptibility to leprosy.
A genetic component to the etiology of leprosy is well recognized but the mechanism of inheritance and the genes involved are yet to be fully established. A genome-wide single nucleotide polymorphism (SNP) based linkage analysis was carried out using 23 pedigrees, each with 3 to 7 family members affected by leprosy. Multipoint parametric and non-parametric linkage analyses were performed using MERLIN 1.1.1. Genome-wide significant evidence for linkage was identified on chromosome 2p14 with a heterogeneity logarithm of odds (HLOD) score of 3.51 (rs1106577) under a recessive model of inheritance, while suggestive evidence was identified on chr.4q22 (HLOD 2.92, rs1349350, dominant model), chr. 8q24 (HLOD 2.74, rs1618523, recessive model) and chr.16q24 (HLOD 1.93, rs276990 dominant model). Our study also provided moderate evidence for a linkage locus on chromosome 6q24-26 by non-parametric linkage analysis (rs6570858, LOD 1.54, p = 0.004), overlapping a previously reported linkage region on chromosome 6q25-26. A genome-wide linkage analysis has identified a new linkage locus on chromosome 2p14 for leprosy in Pedigrees from China.


Nitrated fatty acid, 10-nitrooleate protects against hyperoxia-induced acute lung injury in mice.
The antioxidant and anti-inflammatory effects of electrophilic nitrated fatty acid (NFA); 10-nitrooleate, have been reported. The present study investigated whether 10-nitrooleate has a protective role against hyperoxic-induced acute lung injury (HALI). Using a C57BL/6 mice model of HALI, we investigated the protective effect of 10-nitrooleate. C57BL/6 mice were administered with NFA intratracheally, exposed to hyperoxia for 48 h to induce HALI, and kept at room air for 24 h. Bronchoalveolar lavage (BAL) fluid and lung samples were collected after 24 h of post hyperoxia to analyze markers associated with HALI. Intratracheal (IT) and intraperitoneal (IP) administration of NFA notably attenuated hyperoxia-induced infiltration of inflammatory cells, alveolar-capillary leakage, upregulation of proinflammatory cytokine levels (IL-6 and TNFα) into the BAL fluid, and resolution of inflammation in the lung. Western blot analyses showed that 10-nitrooleate reduced the expression of the inflammatory transcription factor NFκB p65 subunit and increased antioxidant proteins HO-1 and NQO1 expression in the lung tissues compared to vehicle-treated animals. Moreover, 10-nitrooleate reversed the hyperoxia-induced expression of mitophagy-associated markers (PINK1 and p62/SQSTM1), thereby protecting the HALI/ acute respiratory distress syndrome (ARDS). IT and IP delivery of 10-nitrooleate reduces hyperoxia-induced ALI/ARDS by regulating the antioxidant pathways and restoring the mitochondrial homeostasis by regulating mitophagy. It is suggested that NFAs can be further evaluated as supplementary therapy for critically ill patients like COVID-19/ARDS.


A national study of the complications of lupus in pregnancy.
This study was undertaken to determine the risk of rare complications during pregnancy for women with systemic lupus erythematosus. By using the Nationwide Inpatient Sample from 2000-2003, we compared maternal and pregnancy complications for all pregnancy-related admissions for women with and without systemic lupus erythematosus. Of more than 16.7 million admissions for childbirth over the 4 years, 13,555 were to women with systemic lupus erythematosus. Maternal mortality was 20-fold higher among women with systemic lupus erythematosus. The risks for thrombosis, infection, thrombocytopenia, and transfusion were each 3- to 7-fold higher for women with systemic lupus erythematosus. Lupus patients also had a higher risk for cesarean sections (odds ratio: 1.7), preterm labor (odds ratio: 2.4), and preeclampsia (odds ratio: 3.0) than other women. Women with systemic lupus erythematosus were more likely to have other medical conditions, including diabetes, hypertension, and thrombophilia, that are associated with adverse pregnancy outcomes. Women with systemic lupus erythematosus are at increased risk for serious medical and pregnancy complications during pregnancy.


Evaluating the preceptor role for pre-registration nursing and midwifery student clinical education.
The aim of this research is to evaluate the perceptions of the Registered Nurse (RN), Registered Midwife (RM) and Enrolled Nurse (EN) about their experience of preceptoring an undergraduate student within a large Local Health District in New South Wales (NSW) Australia. In the current Health Workforce Australia (HWA) literature, the term 'Clinical Supervisor' has subsumed the role of mentor, preceptor, buddy and facilitator of clinical practice. Preceptor in this paper describes the supervisory, facilitating and teaching role of the registered nurse in the clinical practice undergraduate nursing and midwifery educational pairing. A quantitative cross sectional design was used and data collected using the Clinical Preceptor Experience Evaluation Tool (CPEET), a previously validated and reliable survey tool. Nurses and Midwives across nine acute care facilities that preceptor undergraduate students were invited to complete the survey between March and May 2012. There were 337 survey respondents across nine acute hospitals included in this study (22.5% response rate). Differences were observed between preceptors who had training in precepting in three of the subscales. Differences were observed in all four subscales between those preceptors with access to university facilitators in their location and those without immediate access. The majority of preceptors score highly on all subscales indicating they are generally satisfied with the role of precepting. Significant differences on several items suggest that some aspects of the role are more challenging and less satisfying than others.


[Chondromyxoid fibroma of the mandible: a case report].
Chondromyxoid fibroma (CMF) is a rare benign bone tumor. This tumor mostly affects the long bones of the appendicular skeleton but rarely grows in the craniofacial region. In this article, a case of CMF of the mandible was presented to enhance our understanding of CMF. Its clinical manifestations, imaging characteristics, and treatment methods were discussed by analyzing the related literature.


Fine particulate matter air pollution and subclinical cardiovascular outcomes: A longitudinal study in 15 Chinese cities.
Although previous studies have linked short-term exposure to fine particulate matter (PM2.5) air pollution with various molecular biomarkers of cardiovascular system, limited evidence is available for indicators at clinical or subclinical levels. We examined the associations between short-term PM2.5 exposure and a range of clinical or subclinical indicators of cardiovascular health in general population. A longitudinal repeated-measure study was conducted among 247,640 participants who repeatedly visited health examination centers in 15 typical cities across China from 2013 to 2020. A total of 19 well-established indicators of cardiovascular risk or injury were evaluated and air quality data at nearest fixed-site monitors were collected. Linear mixed-effects models with distributed lag models were used to analyze the potentially lagged effects of PM2.5. The average daily PM2.5 concentration was 48 μg/m3 during the study period. PM2.5 exposure was associated with significant changes of 16 indicators with the effects generally peaked on lag 0 to 3 day. For an interquartile range (IQR) elevation (37 μg/m3) in PM2.5 concentrations over lag 0-7 day, the cumulative percentage changes were 0.50% to 1.27% in heart rates and blood pressure, 0.10% to 5.04% in inflammatory markers, -0.29% to 1.39% in blood viscosity parameters, -0.67% to 3.45% in blood lipids, 0.89% in blood homocysteine, 0.13% to 0.78% in myocardial enzymes, and 3.03% in pulse wave velocity. These associations were not substantially changed after adjusting concomitant exposures to gaseous pollutants. Short-term exposure to PM2.5 may induce early cardiovascular effects in general population, including acute inflammation, myocardial injury, increased blood viscosity, vascular stiffness and hyperlipidemia.


Studies on the surface structure of very low density lipo-proteins.
Very Low Density Lipoproteins (VLDL) were incubated with 5 different pure phospholipases. From the results the following conclusions were drawn. (1) The phospholipids ,re localized in a monomolecular layer on the outside of the VLDL particles. This supports the lipid core model proposed by several groups. (2) A minor fraction of the phospholipids (ranging from 3 to 10%) cannot be degraded by the enzymes and probably have a strong interaction with apoproteins. (3) The average surface pressure of VLDL is probably low, and comparable with a lateral surface pressure of 15 dynes/cm at the air--water interface, as concluded from experiments with two phospholipases A(2). Calculations of the thickness of the surface coat and protein coverage on the basis of this value agree very well with values reported in literature.


The Genetic Basis of Dormancy and Awakening in Cutaneous Metastatic Melanoma.
Immune dysregulation, in combination with genetic and epigenetic alterations, induces an excessive proliferation of uncontrolled melanoma cells followed by dissemination of the tumor cells to distant sites, invading organs and creating metastasis. Although immunotherapy, checkpoint inhibitors and molecular targeted therapies have been developed as treatment options for advanced melanoma, there are specific mechanisms by which cancer cells can escape treatment. One of the main factors associated with reduced response to therapy is the ability of residual tumor cells to persist in a dormant state, without proliferation. This comprehensive review aimed at understanding the genetic basis of dormancy/awakening phenomenon in metastatic melanoma will help identify the possible therapeutical strategies that might eliminate melanoma circulating tumor cells (CTCs) or keep them in the dormant state forever, thereby repressing tumor relapse and metastatic spread.


Morphometry characterisation of European eel spermatozoa with computer-assisted spermatozoa analysis and scanning electron microscopy.
The aim of the present study was to characterise European eel spermatozoa morphometrically, as a basis for future studies on the morphological effects of methods for sperm cryopreservation and sperm quality. This characterisation was carried out measuring several spermatozoa morphology parameters (head length, width, area and perimeter) by scanning electron microscopy (SEM), in comparison with measurements developed in European eel spermatozoa with computer-assisted morphology analysis (ASMA). Spermatozoa head morphology showed differences in width (1.15+/-0.01 microm versus 1.12+/-0.01 microm), perimeter (14.68+/-0.13 microm versus 13.72+/-0.19 microm) and area (5.36+/-0.06 microm2 versus 1.12+/-0.01 microm2) for ASMA and SEM, respectively. When head length was evaluated, significant differences were found, being higher for SEM methodology (5.09+/-0.04 microm versus 4.29+/-0.03 microm). The curved and elongated spermatozoa head in eels means a problem for the ASMA system (Sperm Class Analyser), Morfo Version 1.1, Imagesp, Barcelona, Spain), causing an error in the length measurements. However, similar results were obtained by both techniques when spermatozoa head length was considered as the greater length between two points within the object (4.29+/-0.03 microm versus 4.31+/-0.04 microm for ASMA and SEM, respectively). In conclusion, this is one of the first applications of ASMA in fish and the first in this species, and confirms this system as a useful tool with wide applications in future fish spermatozoa studies. Width, perimeter and area could be used as parameters for the spermatozoa morphology evaluation, whereas the length requires a new programming of the Imagesp software.


Corrections to standard state in combustion calorimetry: an update and a web-based tool.
Combustion calorimetry is the predominant method for determination of enthalpies of formation for organic compounds. Both initial and final states of the calorimeter deviate significantly from the standard conditions. Correction of the obtained results to the standard state must be applied as accurately as possible to determine the combustion energy with an acceptable uncertainty, which is typically a few hundredths of a percent. The correction procedures in their current form were introduced in 1956 with simplifications to allow application in a pre-computer era. In this work, the procedures have been updated with respect to both the equations and reference values. The most reliable data sources are identified, and the updated algorithm is presented in the form of a Web-based tool available through the NIST TRC Web site.


Main anatomical features of the M1 segment of the middle cerebral artery: a 3D time-of-flight magnetic resonance angiography at 3 T study.
The purpose of our study was to determine the main anatomical features of the M1 segment of middle cerebral artery (MRA) using a 3D TOF-MRA at 3 T. Reconstructed and post-processed MRA images were independently analysed by two anatomists in order to determine the course patterns, the division patterns and the early cortical branches patterns of the M1 segments. The division patterns were defined as bipode, tripode or other. The ECB were studied according to their number and their distance from the origin of the M1 segment. The interobserver agreement, to determine the division patterns of the M1 segment, was calculated. The division of the M1 segment was bipode in 73% of the MCAs, monopode in 17%, tripode in 9%, and fan-shaped in 1. In 46% of the cases no ECB was found. In the other cases, only 1 ECB was found and it arose from the medium part of the M1 segment. Our results confirm post-mortem microdissection studies and show that strict anatomical criteria may be applied to 3D TOF MRA at 3 T.


A qualitative exploration of the use of player loans to supplement the talent development process of professional footballers in the under 23 age group of English football academies.
A range of literature has contributed to talent development and career transition in professional football. Recently, attention has shifted to the under 23s age group of English football academies, highlighting limitations in the efficacy of this age group in developing professional footballers. Such limitations have led to players going on loan to football clubs in lower divisions to supplement their development; however, we have yet to develop a scholarly understanding of this approach. Therefore, the aim of this study was to investigate the role of an under 23s player loan in developing professional footballers. Using a qualitative design, data were collected through the use of semi-structured interviews with nine participants. Following a process of thematic analysis, findings highlighted the potential of a loan in contributing to a player's physical and psycho-social development. Positive performances whilst on loan were also perceived to be contributing factors in increased first team selection opportunities at the player's parent club. However, an emphasis was also placed on the need to provide consistent pastoral support to under 23s players during their loan period. Finally, more longitudinal methodologies are required to understand the individual and temporal nature of positive and negative loan experiences of under 23s players.


Excitation of Initial Waves by Wind: A Theoretical Model and Its Experimental Verification.
We present a theory of evolution of wind waves in time and space under abruptly applied wind forcing that is experimentally validated in a laboratory wind-wave tank. The model describes qualitatively and quantitatively the complex wave field development from the initial smooth surface to the finite state. The stochastic nature of wind waves is treated by considering an ensemble of coexisting unstable harmonics that grow due to shear flow instability. Breaking limits the wave growth initially; the process is then controlled by fetch-limited growth duration.


Social monogamy in wild owl monkeys (Aotus azarae) of Argentina: the potential influences of resource distribution and ranging patterns.
Using published and new data from a population of monogamous owl monkeys in the Argentinean Chaco, I examine the hypothesis that social monogamy is a default social system imposed upon males because the spatial and/or temporal distribution of resources and females makes it difficult for a single male to defend access to more than one mate. First, I examine a set of predictions on ranging patterns, use of space, and population density. This first section is followed by a second one considering predictions related to the abundance and distribution of food. Finally, I conclude with a section attempting to link the ranging and ecological data to demographic and life-history parameters as proxies for reproductive success. In support of the hypothesis, owl monkey species do live at densities (7-64 ind/km(2) ) that are predicted for monogamous species, but groups occupy home ranges and core areas that vary substantially in size, with pronounced overlap of home ranges, but not of core areas. There are strong indications that the availability of food sources in the core areas during the dry season may be of substantial importance for regulating social monogamy in owl monkeys. Finally, none of the proxies for the success of groups were strongly related to the size of the home range or core area. The results I present do not support conclusively any single explanation for the evolution of social monogamy in owl monkeys, but they help us to better understand how it may function. Moreover, the absence of conclusive answers linking ranging, ecology, and reproductive success with the evolution of social monogamy in primates, offer renewed motivation for continuing to explore the evolution of monogamy in owl monkeys.


Gravitational effects on fibroblasts' function in relation to wound healing.
The spaceflight environment imposes risks for maintaining a healthy skin function as the observed delayed wound healing can contribute to increased risks of infection. To counteract delayed wound healing in space, a better understanding of the fibroblasts' reaction to altered gravity levels is needed. In this paper, we describe experiments that were carried out at the Large Diameter Centrifuge located in ESA-ESTEC as part of the ESA Academy 2021 Spin Your Thesis! Campaign. We exposed dermal fibroblasts to a set of altered gravity levels, including transitions between simulated microgravity and hypergravity. The addition of the stress hormone cortisol to the cell culture medium was done to account for possible interaction effects of gravity and cortisol exposure. Results show a main impact of cortisol on the secretion of pro-inflammatory cytokines as well as extracellular matrix proteins. Altered gravity mostly induced a delay in cellular migration and changes in mechanosensitive cell structures. Furthermore, 20 × g hypergravity transitions induced changes in nuclear morphology. These findings provide insights into the effect of gravity transitions on the fibroblasts' function related to wound healing, which may be useful for the development of countermeasures.


Medical Specialists' Perspectives on the Influence of Electronic Medical Record Use on the Quality of Hospital Care: Semistructured Interview Study.
Numerous publications show that electronic medical records (EMRs) may make an important contribution to increasing the quality of care. There are indications that particularly the medical specialist plays an important role in the use of EMRs in hospitals. The aim of this study was to examine how, and by which aspects, the relationship between EMR use and the quality of care in hospitals is influenced according to medical specialists. To answer this question, a qualitative study was conducted in the period of August-October 2018. Semistructured interviews of around 90 min were conducted with 11 medical specialists from 11 different Dutch hospitals. For analysis of the answers, we used a previously published taxonomy of factors that can influence the use of EMRs. The professional experience of the participating medical specialists varied between 5 and 27 years. Using the previously published taxonomy, these medical specialists considered technical barriers the most significant for EMR use. The suboptimal change processes surrounding implementation were also perceived as a major barrier. A final major problem is related to the categories "social" (their relationships with the patients and fellow care providers), "psychological" (based on their personal issues, knowledge, and perceptions), and "time" (the time required to select, implement, and learn how to use EMR systems and subsequently enter data into the system). However, the medical specialists also identified potential technical facilitators, particularly in the assured availability of information to all health care professionals involved in the care of a patient. They see promise in using EMRs for medical decision support to improve the quality of care but consider these possibilities currently lacking. The 11 medical specialists shared positive experiences with EMR use when comparing it to formerly used paper records. The fact that involved health care professionals can access patient data at any time they need is considered important. However, in practice, potential quality improvement lags as long as decision support cannot be applied because of the lack of a fully coded patient record.


Left ventricular assist device implantation in heart failure patients with a left ventricular thrombus.
In this report, we share our experience with left ventricular assist device (LVAD) implantation in cases with a left ventricular (LV) thrombus. Over the 3 years, more than 100 end-stage heart failure cases have been treated with LVAD implantation in our center, including 6 patients with a LV thrombus. Three were detected using preoperative transthoracic echocardiography. Fifty percent of the patients had dilated cardiomyopathy and the remaining cases had an ischemic etiology. Double inotropic support with dopamine and dobutamine was used in all, with 3 drugs with the addition of adrenaline in 2 patients. In 4 cases we implanted the HeartWare Ventricular Assist System (HeartWare, Inc., Miramar, Fla, United States) and in the remaining 2 patients, the Berlin Heart EXCOR ventricular assist device (Berlin Heart AG, Berlin, Germany) for biventricular support. In 1 patient the apical ventriculotomy was extended to remove an intertrabecular thrombosis and ventricular septal surface covered with a dacron patch to minimize the thrombogenic potential. Two patients died due to sepsis and multiorgan failure. None of the patients experienced a neurological event, pump thrombosis, or pump malfunction. Two subjects underwent re-explorations due to hemorrhage. Two candidates underwent successfull transplantation without any evidence of thrombosis in the explanted heart or device. We believe that patients with a LV thrombus and preserved right ventricular function are good candidates for implantation of a LVAD after removing the intracavitary thrombus.


Natural calcification of the prosomatic endosternite in the Phalangiidae (Arachnida:Opiliones).
Extracellular masses of crystals are present in the endosternite of certain opilions (Phalangium, Odiellus, and Leiobunum). X-ray spectrography and secondary-ion analysis have shown them to be rich in calcium but poor in phosphorus and other elements. The associated anion has not been identified, but is most probably organic in nature and perhaps an oxalate.


Impact of foods with health logo on saturated fat, sodium and sugar intake of young Dutch adults.
Health logos are introduced to distinguish foods with 'healthier' nutrient composition from regular foods. In the present study, we evaluated the effects of changed food compositions according to health logo criteria on the intake of saturated fat, sugar and sodium in a Dutch population of young adults. Foods in the Dutch food composition table were evaluated against nutrient criteria for logo eligibility. Three replacement scenarios were compared with the nutrient intake 'as measured' in the Dutch consumption survey. The foods not complying with health logo criteria were replaced either by 'virtual' foods exactly complying with the health logo criteria, with real 2007 market shares (scenario I) and 100% market shares (scenario II), or by existing similar foods with a composition that already complied with the health logo criteria (scenario III). The percentage reduction in nutrient intake with the current 2007 market shares of 'health logo foods' was -2.5% for SFA, 0% for sodium and -1% for sugar. With a 100% market share these reductions would be -10% for SFA, -4% for sodium and -6% for sugar. This may lead to a reduction of -40% for SFA, -23% for sodium and -36% for sugar in the most optimal replacement scenario. With 'health logo foods', available in 2007 and current consumption patterns, small reductions can be achieved for SFA and sugar. For additional reductions, lowering the fat/sodium content of meat (products) towards health logo criteria and drinks without sugar towards limits far below health logo criteria would be the most effective reformulation strategy.


HDL-cholesterol levels in the Multiple Risk Factor Intervention Trial (MRFIT) by the MRFIT Research Group 1,2.
Preliminary data from the Multiple Risk Factor Intervention Trial (MRFIT) have been examined for evidence that the program has an influence on plasma HDL-cholesterol. The overall mean level of this lipoprotein in the initial cohort of 1,084 men was not altered by two years of participation in this risk factor reduction project. However, changes did occur, both upwards and downwards, in some individuals. There were significant negative associations between change in HDL-cholesterol and changes in body mass, VLDL-cholesterol, LDL-cholesterol, and serum thiocyanate (a measure of cigarette smoking exposure); and there was a small positive association with change in reported alcohol intake. Multiple regression analysis revealed each of these associations to be independent of the others. The fat-controlled diet designed to lower total serum cholesterol did not decrease HDL-cholesterol levels. We conclude that conventional risk reduction programs are not likely to lower circulating HDL-cholesterol, and that program components such as weight reduction and smoking cessation may increase the levels.


Demonstration of Crohn's disease tissue-specific proteins by enzyme-linked immunosorbent assay (ELISA).
Theories on the etiology of Crohn's disease have included extrinsic agents and intrinsic bowel wall defects. We sought to determine the presence of immunoreactive antigens specific to Crohn's disease tissue by modifying the enzyme-linked immunosorbent assay. Tissue proteins were extracted from four patients with Crohn's disease and from four normal segments of colon from patients with colonic cancer. These tissue extracts were further purified on Con A Sepharose 4B affinity column. The glycoproteins eluted from this column were adsorbed by polystyrene plates as antigen and tested against 85 sera from patients with Crohn's disease, ulcerative colitis, other diarrheal diseases, and normal subjects. Sera from 48 patients with Crohn's disease showed significantly greater recognition of Crohn's disease tissue glycoproteins than sera from 27 disease controls (P less than 0.0125) and 10 normal subjects. These Crohn's disease sera also showed preferential recognition of glycoproteins extracted from Crohn's disease tissue compared to glycoproteins from normal colonic tissue (P less than 0.0005). The nature of these immunoreactive proteins, whether extrinsic or intrinsic, is not yet known. The ELISA may help in further characterization of Crohn's disease tissue-specific glycoprotein(s) and to develop a clinically useful serological test.


The role of GSTP1 polymorphisms and tobacco smoke exposure in children with acute asthma.
The glutathione S-transferase enzymes (GSTs) play an important role in the detoxification of environmental tobacco smoke (ETS), which contributes to airway inflammation, a key component of asthma. Genetic variation in GST genes may influence individuals' ability to detoxify environmental pollutants. To examine the role of polymorphisms in GSTP1 (Ile105Val and Ala114Val), alone and in combination with ETS exposure, on atopy and asthma severity. GSTP1 Ile105Val and Ala114Val were genotyped and ETS exposure was assessed by parental questionnaire, which was validated by urinary cotinine measurements. Associations between ETS exposure, GSTP1 polymorphisms, and their interaction on atopy and asthma severity were investigated. For the functional GSTP1 105 SNP, those with the Ile/Ile genotype had odds for atopy of 2.77 (p = .054) when assessed by genotype alone, which increased to 9.02 (p = .050) when ETS was included, relative to individuals with other genotypes. Likewise, compared to children with other GSTP1 114 genotypes, those with Ala/Ala genotype had a 5.47-fold (p = .002) increased risk of atopy (p = .020) when assessed by genotype alone, increasing to 9.17-fold when ETS was included. The 105 Ile/Ile individuals all had the AA (105 Ile/Ile and 114 Ala/Ala) haplotype group; therefore, the odds for atopy were the same. Individuals without any *C haplotype (105 Val and 114 Val allele) who were exposed to ETS had a 9.17-fold increased risk of atopy when compared with individuals with at least one *C haplotype and not exposed to ETS (p = .020). There were significant interactions between GSTP1 SNPs, atopy, and ETS exposure in this cohort.


Age-related distance esotropia: Clinical features and therapeutic outcomes.
To describe the clinical characteristics and surgical outcomes of a group of patients with age-related distance esotropia (ARDE). A retrospective study was conducted on a consecutive case series of 16 adult patients diagnosed with ARDE between 2008 and 2015. The clinical features evaluated included mean age and gender, primary position deviations at distance and near, measured in prism dioptres (pd), treatment offered in each case, and post-surgical deviations. Ductions and versions were full, with no evidence of lateral rectus paresis. None of these patients had any obvious underlying neurological disorder, such as, high myopia or thyroid disease. A good result is considered to be the disappearance of diplopia in all positions of gaze. A total of 16 patients (11 females [68.8%]) were identified. The mean age at diagnosis was 78.19±6.77 years. The mean initial esodeviation was 2.25±3.08 pd at near (-4 to +8 pd) and 9.5±4.18 pd at distance (2 to 18 pd). Treatment was not necessary in 5 cases because the symptoms were intermittent or well-tolerated. Of the 11 patients with symptoms, one was corrected with an external base therapeutic prism. Botulinum toxin was administered in another patient, without satisfactory results. Unilateral medial rectus muscle recession was performed on one patient, and unilateral lateral rectus plication on 7 patients, indicating prisms before surgery. One patient refused surgery despite continuous diplopia in far vision. After a mean follow-up of 16.5 months, all operated patients were asymptomatic. Not all patients with ARDE require treatment, as the tolerance to diplopia varies from one subject to another. Both medial rectus weakening and lateral rectus strengthening provides excellent results.


Placebo-controlled continuation treatment with mirtazapine: acute pattern of response predicts relapse.
Pattern of response to antidepressants has been proposed as a method to identify patients whose improvement is more likely due to drug vs those whose improvement on drug is more likely to be a placebo effect. It is hypothesized that those with 'true-drug initial response pattern' are most likely to benefit from continuation treatment. The relationship between acute patterns of response and subsequent placebo-controlled continuation treatment with the antidepressant mirtazapine is examined. A total of 410 outpatients were treated openly with mirtazapine for 8-12 weeks. Patients who remitted in the acute phase were randomized to continue the same dose of mirtazapine or switched to placebo. Acute phase responders were classified as 'placebo initial response pattern' (early responders and nonpersistent responders) and 'true-drug initial response pattern' (delayed and persistent responders). Of those with a 'true-drug initial response pattern,' 10/40 (25.0%) relapsed with continuation mirtazapine, and 23/41 (56.1%) relapsed when switched to placebo. The difference (31.1%) is significant. Of those with a 'placebo initial response pattern,' 5/36 (13.9%) relapsed with continuation mirtazapine, and 12/39 (30.8%) relapsed with placebo substitution. This difference (16.9%) is not statistically significant. Moreover, the relapse rate for 'true-drug initial response pattern' patients switched to placebo (56.1%) was also significantly greater than for 'placebo initial response pattern' patients switched to placebo (30.8%). It has been suggested that patients with late onset and persistence are more likely to have improved because of drug. This hypothesis gains support from this study because of the different relapse rates of 'true-drug' responders on drug and placebo. The low relapse rate for patients with an acute placebo pattern switched to placebo suggests specific drug effect played a smaller role in their initial improvement.


The peritumor microenvironment: physics and immunity.
Cancer initiation and progression drastically alter the microenvironment at the interface between healthy and malignant tissue. This site, termed the peritumor, bears unique physical and immune attributes that together further promote tumor progression through interconnected mechanical signaling and immune activity. In this review, we describe the distinct physical features of the peritumoral microenvironment and link their relationship to immune responses. The peritumor is a region rich in biomarkers and therapeutic targets and thus is a key focus for future cancer research as well as clinical outlooks, particularly to understand and overcome novel mechanisms of immunotherapy resistance.


On the parasitoid complex of butterflies with descriptions of two new species of parasitic wasps (Hymenoptera: Eulophidae) from Goa, India.
In comprehensive rearing of butterflies from Goa, India, an interesting parasitoid complex of wasps and tachinid flies was found. Two new species of parasitic wasps are described and illustrated: Tetrastichus thetisae n. sp. (Hymenoptera: Eulophidae), a gregarious parasitoid reared from the pupa of Curetis thetis (Drury) (Lepidoptera: Lycaenidae) on the host plant Derris sp., and Sympiesis thyrsisae n. sp. (Hymenoptera: Eulophidae), a gregarious parasitoid reared from the caterpillar of Gangara thyrsis (Fabricius) (Lepidoptera: Hesperiidae) on the host plant Cocos nucifera L. Additionally, the following host-parasitoid associations are recorded: Amblypodia anita Hewitson (Lepidoptera: Lycaenidae) with Parapanteles sp. (Hymenoptera: Braconidae); Coladenia indrani (Moore) (Lepidoptera: Hesperiidae) with Sympiesis sp. (Hymenoptera: Eulophidae); Danaus chrysippus L. (Lepidoptera: Nymphalidae) with Sturmia convergens (Wiedemann) (Diptera: Tachinidae); Idea malabarica Moore (Lepidoptera: Nymphalidae) with Brachymeria sp. (Hymenoptera: Chalcididae) and Palexorista sp. (Diptera: Tachinidae); Notocrypta curvifascia Felder & Felder (Lepidoptera: Hesperiidae) with Cotesia erionotae (Wilkinson) (Hymenoptera: Braconidae); and Rapala sp. (Lepidoptera: Lycaenidae) with an inominate species close to Aplomya spp. (Diptera: Tachinidae). This discovery is the first record of Tetrastichus as parasitoid of Curetis thetis, Sympiesis as parasitoid of Gangara thyrsis and Coladenia indrani, Brachymeria and Palexorista as parasitoids of Idea malabarica, and Cotesia erionotae as parasitoid of Notocrypta curvifascia. Data on habitat, brief diagnoses and host records for all parasitoids are provided.


Phototherapy in the Evaluation and Management of Photodermatoses.
Ultraviolet light (UV) and visible light are important components in the diagnosis of photodermatoses, and UV has the unique ability to also be used to manage photodermatoses. Phototesting, provocative light testing, and photopatch testing can provide important information in diagnosing patients with photodermatoses; phototesting can be used to determine the starting dose for phototherapy in these patients. Once photosensitivity is established, narrowband UVB and UVA1 therapy have helped to improve the quality of life of photosensitive patients, such as those with polymorphous light eruption, chronic actinic dermatitis, and solar urticaria.


Masked education? The benefits and burdens of wearing face masks in schools during the current Corona pandemic.
Face masks can prevent the spread of the virus SARS-CoV-2, in particular as this spread can occur from people with no symptoms. However, covering the lower half of the face reduces the ability to communicate, interpret, and mimic the expressions of those with whom we interact. Positive emotions become less recognizable, and negative emotions are amplified. Emotional mimicry, contagion, and emotionality in general are reduced and (thereby) bonding between teachers and learners, group cohesion, and learning - of which emotions are a major driver. The benefits and burdens of face masks in schools should be seriously considered and made obvious and clear to teachers and students. The school's specific situation must also inform any decision regarding face mask use.


A Well-Hung Horse: Sired by Knowledge and Imagination.
For more than a century, historians of science have been spinning a philosophical roulette wheel, pondering which is more important in the creative process: imagination or knowledge. The most original scientists (and artists) in our day discover newness by blending existing knowledge with imaginative thinking.


Study of Antioxidant, Antiproliferative and DNA Damage Protecting Activities of Cinnamomum cassia Extracts Obtained by Sequential Extraction.
Cinnamomum cassia (C. cassia) is an evergreen tree in China and Southern and Eastern Asia. In traditional medicine, cinnamon is widely used due to its many bioactivity effects. The present novel study aims to evaluate and make a comparison of antioxidant and antiproliferative activities of different extractions of C. cassia bark using seven solvents having different polarities. Solvents polarity gradients start with the solvent of lower polarity, n-hexane, and end with water as the highest polar solvent. Among the extracts, acetone extract contains the highest phenolic and flavonoid contents; therefore, it is assessed for the ability to protect DNA from damage. The extracts are evaluated for total phenolic, flavonoid contents and antioxidant activities, using FRAP, DPPH, superoxide, and hydroxyl and nitric oxide radicals scavenging assays. DNA damage protecting activity of the acetone extract is studied with the comet assay. Each of the extracts is studied for its antiproliferative effect against, MCF-7, MDA-MB-231(breast cancer), and HT29 (colon cancer), using MTT assay. The acetone extract exhibited the highest FRAP value, phenolic and flavonoids contents when compared to the other extracts and could protect 45% mouse fibroblast cell line (3T3-L1) from DNA damage at 30 μg/ml. The lowest IC50 value in DPPH, superoxide, and hydroxyl radicals scavenging was noticed in the ethyl acetate extract. IC50 value obtained for the hexane extract was the lowest compared to the other extracts in scavenging nitric oxide radicals. The hexane extract showed the highest antiproliferative effect against cancer cells followed by the chloroform extract. The ethyl acetate extract inhibited the proliferation of only MCF-7 by IC50 of 100 μg/ml, while the other extracts exhibited no IC50 in all the cancer cells. C. cassia showed promising antioxidant and anticancer activities with significant DNA damage protecting effect.


Spontaneous breaking of time-reversal symmetry in the pseudogap state of a high-Tc superconductor.
A change in 'symmetry' is often observed when matter undergoes a phase transition-the symmetry is said to be spontaneously broken. The transition made by underdoped high-transition-temperature (high-Tc) superconductors is unusual, in that it is not a mean-field transition as seen in other superconductors. Rather, there is a region in the phase diagram above the superconducting transition temperature Tc (where phase coherence and superconductivity begin) but below a characteristic temperature T* where a 'pseudogap' appears in the spectrum of electronic excitations. It is therefore important to establish if T* is just a cross-over temperature arising from fluctuations in the order parameter that will establish superconductivity at Tc (refs 3, 4), or if it marks a phase transition where symmetry is spontaneously broken. Here we report that, for a material in the pseudogap state, left-circularly polarized photons give a different photocurrent from right-circularly polarized photons. This shows that time-reversal symmetry is spontaneously broken below T*, which therefore corresponds to a phase transition.


Altered muscle activation patterns in symptomatic women during pelvic floor muscle contraction and Valsalva manouevre.
To investigate the muscle activation patterns of the abdomino-pelvic region used by incontinent women during a pelvic floor muscle (PFM) contraction and a Valsalva manouevre compared to healthy, asymptomatic subjects. Thirteen incontinent (symptomatic) women, identified using ultrasound to be consistently depressing the bladder base during PFM contraction, and thirteen continent women (asymptomatic) able to perform an elevating PFM contraction were assessed using surface electromyography (EMG) of the PFM, abdominal and chest wall muscles and vaginal and intra-abdominal pressure (IAP) measurements during PFM contraction and Valsalva under ultrasound surveillance. There were no differences between groups in age, parity or BMI. There was a difference in muscle activation patterns between groups (P = 0.001). During PFM contraction the PFM were less active and the abdominal and chest wall muscles were more active in the symptomatic group. During Valsalva, the PFM and EO were more active in the symptomatic group but there was no difference in the activation of the other muscles between groups. There was a significant interaction (group x pressure) for change in IAP (P = 0.047) but no significant interaction for change in vaginal pressure (VP) (P = 0.324). The symptomatic women displayed altered muscle activation patterns when compared to the asymptomatic group. The symptomatic women were unable to perform a voluntary PFM contraction using a localized muscle strategy, instead activating all the muscles of the abdomino-pelvic cavity. The potential for muscle substitution strategies reinforces the need for close attention to specificity when prescribing PFM exercise programs.


The doctrine of "Reasonable Doctor" in medical negligence: Need to be more reasonable.
The model of a 'reasonable doctor' has been quite successfully used to deliver justice in disputes involving medical negligence. However, many a times a doctor is held guilty of negligence when viewed through the narrow lens of an ideal 'reasonable doctor' and without looking into the circumstances under which he was working which could have actually led to the alleged act of negligence. This short write-up highlights the importance of applying this doctrine more reasonably in the best interest of all stake holders and the drawbacks of the doctrine of 'reasonable doctor' in adjudicating medical negligence cases with few international case laws.


Biopsychosocial issues and risk factors in the family when the child has a chronic illness.
Between 10 and 20 million American children and adolescents have some type of chronic health condition or impairment. There has been a recent interest in the psychological aspects--emotional and behavioral--of chronic illness in children and adolescents. Major new areas of science, such as behavioral medicine, have emerged to address the research and clinical demands of this field. Several studies already have demonstrated the increased risk for psychiatric problems in children with chronic health conditions to be three to four times greater than their healthy peers. The proposed perspective shifts from a traditional sole reliance on medical diagnosis to a focus more on the dimensions of adjustment, socioeconomic status, visibility of condition, social support, and family functioning.


Rapid and sensitive detection of clomazone in potato and pumpkin samples using a gold nanoparticle-based lateral-flow strip.
In this study, an ultrasensitive monoclonal antibody (mAb) was produced and used to develop a gold nanoparticle-based lateral flow immunochromatographic (ICA) strip for screening of clomazone (CLO) in potato and pumpkin samples. With assayed by indirect competitive enzyme-linked immunosorbent assay (ic-ELISA) method, the mAb belonging of IgG2 subclass showed a half-maximal inhibitory concentration (IC50) of 3.47 ng/mL and a linear range of detection of 0.43-28.09 ng/mL. A cross-reactivity test revealed that the mAb had good specificity for CLO. The strip assay had a visual limit of detection (LOD) of 5 µg/kg and a cut-off value of 50 µg/kg for CLO pumpkin samples (potato samples was 100 µg/kg) when evaluated with the naked eye. The results were consistent with ic-ELISA and high performance liquid chromatography tandem mass spectrometry (HPLC-MS). Thus, this ICA strip assay represents a potentially tool for on-site and rapid initial detection of CLO in potato and pumpkin samples.


Pericardiocentesis for symptomatic malignant pericardial effusion: a study of 36 patients.
We reviewed 36 cases of symptomatic malignant pericardial effusion managed with pericardiocentesis at our institution from 1982 to 1989. There were 13 men and 23 women, aged 49 +/- 12 years (range, 33-76 years). The commonest underlying tumours were lung cancer (12 cases, 33%) and breast cancer (11 cases, 30%). Pericardiocentesis was successful as the initial management in 34 of 36 patients (94%); one patient died as a result of the procedure and another required subxiphoid incision and tube drainage of the effusion. When intrapericardial sclerotherapy was performed, only three of 28 patients required repeat pericardiocentesis, and when sclerotherapy was not performed initially, four of seven patients had recurrent symptomatic effusions. Median survival following pericardiocentesis in breast cancer patients was 10 months (range, 0-36 months) and in all other malignancies was four months (range, 0-12 months). We conclude that pericardiocentesis with intrapericardial sclerotherapy provides good local control for symptomatic malignant pericardial effusion in the majority of patients. In spite of this, the median survival of such patients is poor, especially in patients with malignancies other than breast cancer, with few patients surviving more than a few months.


Transcript profiling reveals novel marker genes involved in fruiting body formation in Tuber borchii.
cDNA arrays were used to explore mechanisms controlling fruiting body development in the truffle Tuber borchii. Differences in gene expression were higher between reproductive and vegetative stage than between two stages of fruiting body maturation. We suggest hypotheses about the importance of various physiological processes during the development of fruiting bodies.


Identification of cell context-dependent YAP-associated proteins reveals β1 and β4 integrin mediate YAP translocation independently of cell spreading.
Yes-associated protein (YAP) is a transcriptional regulator and mechanotransducer, relaying extracellular matrix (ECM) stiffness into proliferative gene expression in 2D culture. Previous studies show that YAP activation is dependent on F-actin stress fiber mediated nuclear pore opening, however the protein mediators of YAP translocation remain unclear. Here, we show that YAP co-localizes with F-actin during activating conditions, such as sparse plating and culturing on stiff 2D substrates. To identify proteins mediating YAP translocation, we performed co-immunoprecipitation followed by mass spectrometry (co-IP/MS) for proteins that differentially associated with YAP under activating conditions. Interestingly, YAP preferentially associates with β1 integrin under activating conditions, and β4 integrin under inactivating conditions. In activating conditions, CRISPR/Cas9 knockout (KO) of β1 integrin (ΔITGB1) resulted in decreased cell area, which correlated with decreased YAP nuclear localization. ΔITGB1 did not significantly affect the slope of the correlation between YAP nuclear localization with area, but did decrease overall nuclear YAP independently of cell spreading. In contrast, β4 integrin KO (ΔITGB4) cells showed no change in cell area and similarly decreased nuclear YAP. These results reveal proteins that differentially associate with YAP during activation, which may aid in regulating YAP nuclear translocation.


Antigen-presenting capacity of guinea pig B lymphocytes in T cell proliferative response in vitro.
The capacity of normal (unprimed) B cells and keyhole-limpet-hemocyanin(KLH)-primed B cells to present the antigen KLH to KLH-primed T cells in the guinea pig was examined with in vitro assay of lymphocyte proliferative response. Antigen-pulsed B-lymphocyte population, extensively devoid of macrophages (M phi), induced the proliferative response of KLH-primed T lymphocytes, although less effectively than the peritoneal M phi. The antigen presentation was antigen-specific. There was no substantial difference between the magnitude of T-cell response induced by KLH-primed B-cell population and that stimulated by unprimed, normal B-cell population. The antigen-presenting capacity of the B-cell population was abrogated by pretreatment with anti-guinea pig immunoglobulin (Ig) or anti-I-region-associated (Ia) antigen antiserum and complement, whereas it was not affected with anti-guinea pig M phi antiserum and complement. From studies using strain 2 and strain 13 guinea pigs, histocompatibility requirement between antigen-pulsed B cells and antigen-reactive T cells was suggested: B lymphocytes, as well as M phi, did elicit proliferative response to specific antigen, KLH or purified protein derivative of tuberculin (PPD), in syngeneic, but not in allogeneic, T cells. These results suggest that the Ia-positive, surface-Ig-bearing guinea-pig B lymphocytes - not only KLH-primed B cells but also unprimed B cells - are capable of presenting antigen to primed T cells in a major histocompatibility complex(MHC)-restricted fashion.


Correction: CATALISE: A Multinational and Multidisciplinary Delphi Consensus Study. Identifying Language Impairments in Children.
[This corrects the article DOI: 10.1371/journal.pone.0158753.].


High dose gammaglobulin therapy in adults with idiopathic thrombocytopenic purpura (ITP). Clinical effects.
This study of the effect of high-dose intravenous gammaglobulins with one or two courses of therapy in 18 adults with idiopathic thrombocytopenia purpura showed a platelet rise in thirteen patients. The highest response rates were seen in splenectomized adults. In chronic patients the response was transient only. If therapy was effective, increased values of platelet-associated IgG were reduced, while shortened platelet survival times were prolonged. There was no influence of high-dose gammaglobulins on platelet function. different 7S-preparations such as beta-propiolactone modified Ig, pH 4 treated Ig and reduced and alkylated Ig have comparable effects.


Use of GC × GC/TOF-MS and LC/TOF-MS for metabolomic analysis of Hyalella azteca chronically exposed to atrazine and its primary metabolite, desethylatrazine.
Atrazine is one of the most commonly detected contaminants in the U.S. Little information is available on one of atrazine's metabolites, desethylatrazine (DEA). Two-dimensional gas chromatography and liquid chromatography coupled with time of flight- mass spectrometry were used to examine metabolite profiles of Hyalella azteca chronically exposed to 30 µg/L atrazine and DEA. The majority of identified metabolites were by-products of β-oxidation of fatty acids suggesting possible disruption in energy metabolism. Eicosanoids increased in exposed females suggesting possible perturbations in neuropeptide hormonal systems. Overall, this research demonstrates the feasibility of utilizing metabolomic profiling of invertebrate species exposed to environmental contaminants as a way to determine mechanisms of toxicity.