Enhancing the antigenicity and immunogenicity of monomeric forms of hepatitis C virus E2 for use as a preventive vaccine.
The E2 glycoprotein of hepatitis C virus (HCV) is the major target of broadly neutralizing antibodies (bNAbs) that are critical for the efficacy of a prophylactic HCV vaccine. We previously showed that a cell culture-derived, disulfide-linked high-molecular-weight (HMW) form of the E2 receptor-binding domain lacking three variable regions, Δ123-HMW, elicits broad neutralizing activity against the seven major genotypes of HCV. A limitation to the use of this antigen is that it is produced only at low yields and does not have a homogeneous composition. Here, we employed a sequential reduction and oxidation strategy to efficiently refold two high-yielding monomeric E2 species, D123 and a disulfide-minimized version (D123A7), into disulfide-linked HMW-like species (Δ123r and Δ123A7r). These proteins exhibited normal reactivity to bNAbs with continuous epitopes on the neutralizing face of E2, but reduced reactivity to conformation-dependent bNAbs and nonneutralizing antibodies (non-NAbs) compared with the corresponding monomeric species. Δ123r and Δ123A7r recapitulated the immunogenic properties of cell culture-derived D123-HMW in guinea pigs. The refolded antigens elicited antibodies that neutralized homologous and heterologous HCV genotypes, blocked the interaction between E2 and its cellular receptor CD81, and targeted the AS412, AS434, and AR3 domains. Of note, antibodies directed to epitopes overlapping with those of non-NAbs were absent. The approach to E2 antigen engineering outlined here provides an avenue for the development of preventive HCV vaccine candidates that induce bNAbs at higher yield and lower cost.


Evaluation and management of emergencies in patients with mechanical circulatory support devices.
Mechanical circulatory support devices such as ventricular assist devices have become the approved and accepted treatment option to improve survival and quality of life in patients with advanced heart failure refractory to medical therapy. Patients as a result are living longer and presenting to emergency medical services, primary care facilities, emergency departments, and critical care units more frequently. Currently, health care providers have limited experience in managing this complex patient population. Thus, we created a standardized method of initial evaluation and algorithmic approach of management to help in the management of emergencies in this unique patient population. We present a comprehensive overview of the types of mechanical assist devices, their complications, and an algorithmic approach to the emergency management in the patients with mechanical circulatory support. A systematic method is crucial in prompt and early recognition of emergencies to ensure appropriate management of these patients.


Quest to develop a standard screening method for urothelial carcinoma using liquid-based cytology (The Paris System) and CK20.
Bladder cancer, the most common malignancy of the urinary tract is a leading cause of morbidity and mortality. But cystoscopy, which is till now the mainstay of screening, is an invasive, high-cost method with low sensitivity especially for flat lesions. To find a non-invasive and effective screening method with liquid-based cytology (LBC) using The Paris System (TPS) and CK20 immunocytochemistry. It was a prospective study including the patients with clinical or cystoscopic diagnosis of urinary bladder space occupying lesions (SOL). Both conventional (CC) and liquid-based cytology slides were prepared from urine samples. Slides were evaluated by two trained pathologists and categorized according to TPS guidelines. CK20 immunocytochemistry (ICC) was also performed. Consequent formalin-fixed paraffin embedded sections were blindly examined by another pathologist and was taken as gold standard for comparison. All the statistical analysis were done using MedCalc version 15.8 [Mariakerke, Belgium: MedCalc Software 2015]. The study included 150 cases with a mean age of 62.4 years. Five cases revealed unsatisfactory smears. Rest of the cases were categorized as the following: 18.1% as NH-GUC, 8% as LGUN, 22.1% as AUC, 15.4% as SH-GUC, 32.9% as HGUC. Kappa value of CC and LBC were strong (0.854). LBC alone showed very low specificity (58%) and PPV (74.8%) which improved on application of ICC (specificity: 97.4%, PPV: 96.3%). We conclude that CK20 ICC offers potential for accurate, non-invasive detection and surveillance of bladder cancer and is a better tool when combined with liquid-based cytology, reported using The Paris System.


Analyzing the Effects of Topological Defect (TD) on the Energy Spectra and Thermal Properties of LiH, TiC and I2 Diatomic Molecules.
In this study, the impacts of TD on the energy spectra and thermal properties of LiH, TiC and I2 diatomic molecules is considered. The Schrodinger equation in cosmic string spacetime is solved with the generalized Morse potential using the well-known (NU) method. The energy spectra and eigenfunction are obtained respectively. The energy spectra is used to obtain the partition function which is then used to evaluate the thermal properties of the system is evaluated accordingly. We find that the energy spectra in the presence of the TD differ from their flat Minkowski spacetime analogue. The effects of the deformation parameter and TD on the thermal properties of the system is also analysed in detail. We observe that the specific heat capacity of the system tends to exhibit quasi-saturation as the deformation parameter and topological defect approaches unity. The results of our study can be applied in the astrophysical situation where these modifications exist in the understanding of spectroscopical data and it may be used as a probe of the presence of a cosmic string or a global monopole in the Universe.


Does speed of intrathecal injection affect the distribution of 0.5% hyperbaric bupivacaine?
We have evaluated the influence of speed of intrathecal injection on lateral distribution of 0.5% hyperbaric bupivacaine. We studied 60 patients undergoing lower limb surgery who were placed in the lateral position with the operative side in the dependent position. After dural puncture (25-gauge Whitacre spinal needle), the needle aperture was turned towards the dependent side and 0.5% hyperbaric bupivacaine 8 mg was injected randomly at a rate of 0.02 ml s-1 (group slow, n = 30) or 0.25 ml s-1 (group fast, n = 30). Lateral position was maintained for 15 min while a blinded observer recorded loss of pinprick sensation and degree of motor block on both surgical and non-surgical sides. There were no differences between the groups. Forty-five minutes after patients were turned to the supine position, spinal anaesthesia was unilateral in 17 patients in group slow (56%) and in 13 patients in group fast (43%). We conclude that using extremely low speeds for intrathecal injection were not clinically advantageous in obtaining unilateral spinal anaesthesia.


Hepatitis B virus (HBV) and hepatitis C virus (HCV) infections in health care workers (HCWs): guidelines for prevention of transmission of HBV and HCV from HCW to patients.
The transmission of viral hepatitis from health care workers (HCW) to patients is of worldwide concern. Since the introduction of serologic testing in the 1970s there have been over 45 reports of hepatitis B virus (HBV) transmission from HCW to patients, which have resulted in more than 400 infected patients. In addition there are six published reports of transmissions of hepatitis C virus (HCV) from HCW to patients resulting in the infection of 14 patients. Additional HCV cases are known of in the US and UK, but unpublished. At present the guidelines for preventing HCW to patient transmission of viral hepatitis vary greatly between countries. It was our aim to reach a Europe-wide consensus on this issue. In order to do this, experts in blood-borne infection, from 16 countries, were questioned on their national protocols. The replies given by participating countries formed the basis of a discussion document. This paper was then discussed at a meeting with each of the participating countries in order to reach a Europe-wide consensus on the identification of infected HCWs, protection of susceptible HCWs, management and treatment options for the infected HCW. The results of that process are discussed and recommendations formed. The guidelines produced aim to reduce the risk of transmission from infected HCWs to patients. The document is designed to complement existing guidelines or form the basis for the development of new guidelines. This guidance is applicable to all HCWs who perform EPP, whether newly appointed or already in post.


The IASP classification of chronic pain for ICD-11: chronic primary pain.
This article describes a proposal for the new diagnosis of chronic primary pain (CPP) in ICD-11. Chronic primary pain is chosen when pain has persisted for more than 3 months and is associated with significant emotional distress and/or functional disability, and the pain is not better accounted for by another condition. As with all pain, the article assumes a biopsychosocial framework for understanding CPP, which means all subtypes of the diagnosis are considered to be multifactorial in nature, with biological, psychological, and social factors contributing to each. Unlike the perspectives found in DSM-5 and ICD-10, the diagnosis of CPP is considered to be appropriate independently of identified biological or psychological contributors, unless another diagnosis would better account for the presenting symptoms. Such other diagnoses are called "chronic secondary pain" where pain may at least initially be conceived as a symptom secondary to an underlying disease. The goal here is to create a classification that is useful in both primary care and specialized pain management settings for the development of individualized management plans, and to assist both clinicians and researchers by providing a more accurate description of each diagnostic category.


Replication of the R factor Rts1 in Proteus mirabilis.
The replication of the R factor Rts1 in Proteus mirabilis was examined by using the technique of CsCl density-gradient centrifugation. The proportion of Rts1 deoxyribonucleic acid (DNA) relative to the host chromosomal DNA (% R-DNA) was 7% in both exponential and stationary growth phases in Penassay Broth and supplemented M9 minimal medium at 30 C. The chromosomal DNA content per cell varied over a threefold range in the different growth media. In agreement with previous genetic observations, the replication of Rts1 was found to be temperature-sensitive and Rts1 DNA was diluted from the cells during exponential growth at 42 C. (14)N-(15)N medium transfer experiments have shown that individual copies of Rts1 are selected at random for replication during the duplication of the multicopy episome pool.


Area deprivation across the life course and physical capability in midlife: findings from the 1946 British Birth cohort.
Physical capability in later life is influenced by factors occurring across the life course, yet exposures to area conditions have only been examined cross-sectionally. Data from the National Survey of Health and Development, a longitudinal study of a 1946 British birth cohort, were used to estimate associations of area deprivation (defined as percentage of employed people working in partly skilled or unskilled occupations) at ages 4, 26, and 53 years (residential addresses linked to census data in 1950, 1972, and 1999) with 3 measures of physical capability at age 53 years: grip strength, standing balance, and chair-rise time. Cross-classified multilevel models with individuals nested within areas at the 3 ages showed that models assessing a single time point underestimate total area contributions to physical capability. For balance and chair-rise performance, associations with area deprivation in midlife were robust to adjustment for individual socioeconomic position and prior area deprivation (mean change for a 1-standard-deviation increase: balance, -7.4% (95% confidence interval (CI): -12.8, -2.8); chair rise, 2.1% (95% CI: -0.1, 4.3)). In addition, area deprivation in childhood was related to balance after adjustment for childhood socioeconomic position (-5.1%, 95% CI: -8.7, -1.6). Interventions aimed at reducing midlife disparities in physical capability should target the socioeconomic environment of individuals-for standing balance, as early as childhood.


A qualitative investigation of the impact that therapeutic recreational camps have on the psychological wellbeing of siblings of individuals with health conditions.
The siblings of individuals with health conditions are often overlooked, despite being at risk of experiencing psychological difficulties. There is a lack of literature investigating interventions which could promote siblings' psychological wellbeing. Therapeutic recreational (TR) camps promote self-perception and self-worth, yet there are currently no UK studies qualitatively investigating siblings' experiences of TR camps. This study aims to understand siblings' lived experiences of attending TR camps, providing a greater depth of understanding of whether these camps have a positive impact on siblings' wellbeing. Due to the paucity of qualitative research regarding individuals' experiences of TR camps, semi-structured interviews were conducted with eight individuals who had attended TR sibling camps. Verbatim transcripts were analysed using Interpretative Phenomenological Analysis (IPA). Analysis revealed three superordinate themes. The first, an environment that facilitates autonomous challenge, demonstrated the importance of siblings having a place where they felt autonomous in a fun, care-free environment. This encouraged participants to engage in challenges, feeling determined to overcome them. The second, an inclusive and supportive environment, highlighted the importance of connectedness. Facing challenges and reflecting on this within a supportive team, encouraged a sense of belonging. The final superordinate theme, a transformational journey, revealed that siblings gained determination and a positive outlook to facing challenges in daily life. Additionally, they gained self-acceptance and a positive self-image, becoming more confident with, and accepting of, their identity. This study presents a novel contribution to the existing literature and highlights the importance of camp providers ensuring their TR model incorporates autonomous challenge, success and reflection. These encourage positive youth development and self-determination in young people. In addition, recommendations include implementing teams, promoting positive volunteer-camper relationships and implementing reflective sessions. These recommendations could potentially enhance the positive impact that TR camps have on siblings' psychological wellbeing.


Eccentric exercise markedly increases c-Jun NH(2)-terminal kinase activity in human skeletal muscle.
Eccentric contractions require the lengthening of skeletal muscle during force production and result in acute and prolonged muscle injury. Because a variety of stressors, including physical exercise and injury, can result in the activation of the c-Jun NH(2)-terminal kinase (JNK) intracellular signaling cascade in skeletal muscle, we investigated the effects of eccentric exercise on the activation of this stress-activated protein kinase in human skeletal muscle. Twelve healthy subjects (7 men, 5 women) completed maximal concentric or eccentric knee extensions on a KinCom isokinetic dynamometer (10 sets, 10 repetitions). Percutaneous needle biopsies were obtained from the vastus lateralis muscle 24 h before exercise (basal), immediately postexercise, and 6 h postexercise. Whereas both forms of exercise increased JNK activity immediately postexercise, eccentric contractions resulted in a much higher activation (15.4 +/- 4.5 vs. 3.5 +/- 1.4-fold increase above basal, eccentric vs. concentric). By 6 h after exercise, JNK activity decreased back to baseline values. In contrast to the greater activation of JNK with eccentric exercise, the mitogen-activated protein kinase kinase 4, the immediate upstream regulator of JNK, was similarly activated by concentric and eccentric exercise. Because the activation of JNK promotes the phosphorylation of a variety of transcription factors, including c-Jun, the results from this study suggest that JNK may be involved in the molecular and cellular adaptations that occur in response to injury-producing exercise in human skeletal muscle.


Prescribing Practices of Quetiapine for Insomnia at a Tertiary Care Inpatient Child and Adolescent Psychiatry Unit: A Continuous Quality Improvement Project.
To examine the prescribing practices of quetiapine for insomnia at a tertiary care child and adolescent psychiatric inpatient unit. A retrospective chart review was conducted on all admissions in 2013 involving night-time only prescription of quetiapine. We examined patient demographics, discharge diagnoses, physician's written indications for prescriptions, and maximum doses used. If used for insomnia only, we noted any documentation of past sedative trials, concurrent prescriptions of other sedative agents, whether quetiapine was started in hospital or continued as a part of a community regimen, and whether quetiapine was continued on discharge. Of 720 admissions, 83 (11.5%) involved the prescription of night-time only quetiapine, and 47 of the 83 (57%) were for insomnia only. Of patients prescribed quetiapine for insomnia only, most common discharge diagnoses were anxiety disorder (35%), depressive disorder (27%), eating disorder (27%), and Cluster B/borderline personality traits/disorder (25%). Mean age was 15.4 years; mean maximum dose was 41.2 mg. Quetiapine was often started during admission (89.5%) and continued on discharge (66%). About 40% of these cases involved concurrent prescription of other sedative agents. Most patients (81%) had no documented history of prior sedative trials. Quetiapine is used not infrequently for the management of insomnia in adolescents in tertiary mental health settings. We highlight the nuances associated with the prescription of quetiapine for the treatment of insomnia in the unique setting of the child and adolescent psychiatric inpatient unit, emphasizing the importance of weighing short-term use with potential long-term adverse consequences if continued in the community setting.


Resolving the fibrotic niche of human liver cirrhosis at single-cell level.
Liver cirrhosis is a major cause of death worldwide and is characterized by extensive fibrosis. There are currently no effective antifibrotic therapies available. To obtain a better understanding of the cellular and molecular mechanisms involved in disease pathogenesis and enable the discovery of therapeutic targets, here we profile the transcriptomes of more than 100,000 single human cells, yielding molecular definitions for non-parenchymal cell types that are found in healthy and cirrhotic human liver. We identify a scar-associated TREM2+CD9+ subpopulation of macrophages, which expands in liver fibrosis, differentiates from circulating monocytes and is pro-fibrogenic. We also define ACKR1+ and PLVAP+ endothelial cells that expand in cirrhosis, are topographically restricted to the fibrotic niche and enhance the transmigration of leucocytes. Multi-lineage modelling of ligand and receptor interactions between the scar-associated macrophages, endothelial cells and PDGFRα+ collagen-producing mesenchymal cells reveals intra-scar activity of several pro-fibrogenic pathways including TNFRSF12A, PDGFR and NOTCH signalling. Our work dissects unanticipated aspects of the cellular and molecular basis of human organ fibrosis at a single-cell level, and provides a conceptual framework for the discovery of rational therapeutic targets in liver cirrhosis.


Plasma adiponectin levels and risk of heart failure, atrial fibrillation, aortic valve stenosis, and myocardial infarction: large scale observational and Mendelian randomization evidence.
Adiponectin may play an important protective role in heart failure and associated cardiovascular diseases. We hypothesized that plasma adiponectin is associated observationally and causally, genetically with risk of heart failure, atrial fibrillation, aortic valve stenosis, and myocardial infarction. In the Copenhagen General Population Study, we examined 30,045 individuals with plasma adiponectin measurements observationally and 96,903 individuals genetically in one-sample Mendelian randomization analyses using five genetic variants explaining 3% of the variation in plasma adiponectin. In the HERMES, UK Biobank, The Nord-Trøndelag Health Study(HUNT), deCODE, the Michigan Genomics Initiative(MGI), DiscovEHR, and the AFGen consortia, we performed two-sample Mendelian randomization analyses in up to 1,030,836 individuals using 12 genetic variants explaining 14% of the variation in plasma adiponectin.In observational analyses modelled linearly, a 1 unit log-transformed higher plasma adiponectin was associated with a hazard ratio of 1.51(95% confidence interval:1.37-1.66) for heart failure, 1.63(1.50-1.78) for atrial fibrillation, 1.21(1.03-1.41) for aortic valve stenosis, and 1.03(0.93-1.14) for myocardial infarction; levels above the median were also associated with increased risk of myocardial infarction, and non-linear U-shaped associations were more apparent for heart failure, aortic valve stenosis, and myocardial infarction in less-adjusted models. Corresponding genetic, causal risk ratios were 0.92(0.65-1.29), 0.87(0.68-1.12), 1.55(0.87-2.76), and 0.93(0.67-1.30) in one-sample Mendelian randomization analyses, and no significant associations were seen for non-linear one-sample Mendelian randomization analyses; corresponding causal risk ratios were 0.99(0.89-1.09), 1.00(0.92-1.08), 1.01(0.79-1.28), and 0.99(0.86-1.13) in two-sample Mendelian randomization analyses, respectively. Observationally, elevated plasma adiponectin was associated with increased risk of heart failure, atrial fibrillation, aortic valve stenosis, and myocardial infarction. However, genetic evidence did not support causality for these associations.


Aldehyde dehydrogenase 1A1 increases NADH levels and promotes tumor growth via glutathione/dihydrolipoic acid-dependent NAD+ reduction.
Aldehyde dehydrogenase 1A1 (ALDH1A1) is a member of the aldehyde dehydrogenase superfamily that oxidizes aldehydes to their corresponding acids, reactions that are coupled to the reduction of NAD+ to NADH. We report here that ALDH1A1 can also use glutathione (GSH) and dihydrolipoic acid (DHLA) as electron donors to reduce NAD+ to NADH. The GSH/DHLA-dependent NAD+-reduction activity of ALDH1A1 is not affected by the aldehyde dehydrogenase inhibitor or by mutation of the residues in its aldehyde-binding pocket. It is thus a distinct biochemical reaction from the classic aldehyde-dehydrogenase activity catalyzed by ALDH1A1. We also found that the ectopic expression of ALDH1A1 decreased the intracellular NAD+/NADH ratio, while knockout of ALDH1A1 increased the NAD+/NADH ratio. Simultaneous knockout of ALDH1A1 and its isozyme ALDH3A1 in lung cancer cell line NCI-H460 inhibited tumor growth in a xenograft model. Moreover, the ALDH1A1 mutants that retained their GSH/DHLA-dependent NAD+ reduction activity but lost their aldehyde-dehydrogenase activity were able to decrease the NAD+/NADH ratio and to rescue the impaired growth of ALDH1A1/3A1 double knockout tumor cells. Collectively, these results suggest that this newly characterized GSH/DHLA-dependent NAD+-reduction activity of ALDH1A1 can decrease cellular NAD+/NADH ratio and promote tumor growth.


Intracellular Ca(2+) and TRPM5-mediated membrane depolarization produce ATP secretion from taste receptor cells.
ATP is a transmitter secreted from taste bud receptor (Type II) cells through ATP-permeable gap junction hemichannels most probably composed of pannexin 1. The elevation of intracellular Ca(2+) and membrane depolarization are both believed to be involved in transmitter secretion from receptor cells, but their specific roles have not been fully elucidated. In the present study, we show that taste-evoked ATP secretion from mouse vallate receptor cells is evoked by the combination of intracellular Ca(2+) release and membrane depolarization. Unexpectedly, ATP secretion is not blocked by tetrodotoxin, indicating that transmitter release from these cells still takes place in the absence of action potentials. Taste-evoked ATP secretion is absent in receptor cells isolated from TRPM5 knockout mice or in taste cells from wild type mice where current through TRPM5 channels has been eliminated. These findings suggest that membrane voltage initiated by TRPM5 channels is required for ATP secretion during taste reception. Nonetheless, even in the absence of TRPM5 channel activity, ATP release could be triggered by depolarizing cells with KCl. Collectively, the findings indicate that taste-evoked elevation of intracellular Ca(2+) has a dual role: (1) Ca(2+) opens TRPM5 channels to depolarize receptor cells and (2) Ca(2+) plus membrane depolarization opens ATP-permeable gap junction hemichannels.


Transient 2D IR spectroscopy of charge injection in dye-sensitized nanocrystalline thin films.
We use nonlinear 2D IR spectroscopy to study TiO(2) nanocrystalline thin films sensitized with a Re dye. We find that the free electron signal, which often obscures the vibrational features in the transient absorption spectrum, is not observed in the 2D IR spectra. Its absence allows the vibrational features of the dye to be much better resolved than with the typical IR absorption probe. We observe multiple absorption bands but no cross peaks in the 2D IR spectra, which indicates that the dyes have at least three conformations. Furthermore, by using a pulse sequence in which we initiate electron transfer in the middle of the infrared pulse train, we are able to assign the excited state features by correlating them to the ground state vibrational modes and determine that the three conformations have different time scales and cross sections for electron injection. 2D IR spectroscopy is proving to be very useful in disentangling overlapping structural distributions in biological and chemical physics processes. These experiments demonstrate that nonlinear infrared probes are also a powerful new tool for studying charge transfer at interfaces.


The utility of MASS-FIX to detect and monitor monoclonal proteins in the clinic.
The detection and quantification of monoclonal-proteins (M-proteins) are necessary for the diagnosis and evaluation of response in plasma cell dyscrasias. Immunoglobulin enrichment-coupled with matrix-assisted laser desorption ionization time-of-flight mass-spectrometry (MASS-FIX) is a simple and inexpensive method to identify M-proteins, but its clinical generalizability has not yet been elucidated. We compared MASS-FIX to protein electrophoresis (PEL), serum/urine immunofixation-electrophoresis (IFE), and quantitative serum free-light chain (FLC) for the identification of M-proteins in different clinical diagnoses. Paired serum and urine samples from 257 patients were tested. There were six patients for whom s-IFE and FLC ratio were positive and serum MASS-FIX was negative, but when serum and urine MASS-FIX results were combined, only one patient with light chain-MGUS was missed. Serum/urine-MASS-FIX detected M-proteins in 18 patients with negative serum/urine-PEL/IFE and serum-FLC, 10 of whom had multiple myeloma or AL amyloidosis, who were mistakenly thought to have complete hematologic response by serum/urine-PEL/IFE and serum-FLC. Nearly half of the AL amyloidosis patients had atypical spectra, which may prove to be a clue to the diagnosis and pathogenesis of the disease. In conclusion, MASS-FIX has a comparable sensitivity with PEL/IFE/FLC methods and can help inform the clinical diagnosis.


Is Short-Term Exposure to PM2.5 Relevant to Childhood Kawasaki Disease?
Background: Kawasaki disease (KD) is an acute febrile vascular disease of unknown cause that affects the whole body. KD typically occurs in infants under the age of five and is found mainly in East Asian countries. Few studies have reported on the relationship between the pollutant PM2.5 and KD, and the evidence remains irrelevant or insufficient. Objectives: We investigated the relationship between short-term exposure to PM2.5 and KD hospitalizations using data from Ewha Womans University Mokdong Hospital, 2006 to 2016. Methods: We obtained data from the hospital EMR (electronic medical records) system. We evaluated the relationship between short-term exposure to PM2.5 and KD hospitalizations using a case-crossover design. We considered exposures to PM2.5 two weeks before the date of KD hospitalization. We analyzed the data using a conditional logistic regression adjusted for temperature and humidity. The effect size was calculated as a 10 μg/m3 increase in PM2.5 concentration. We performed a subgroup analysis by sex, season, age group, and region. In the two-pollutants model, we adjusted SO2, NO2, CO, and O3, but the effect size did not change. Results: A total of 771 KD cases were included in this study. We did not find any statistically significant relationship between PM2.5 and children's KD hospitalization (two-day moving average: odds ratio (OR) = 1.01, 95% confidence intervals (CI) = 0.95, 1.06; seven-day moving average: OR = 0.98, CI = 0.91, 1.06; 14-day moving average: OR = 0.93, CI = 0.82, 1.05). A subgroup analysis and two pollutant analysis also found no significant results. Conclusion: We did not find a statistically significant relationship between PM2.5 and children's KD hospitalizations. More research is needed to clarify the association between air pollution, including PM2.5, and KD.


Characterization of the repair of injury induced by freezing Salmonella anatum.
Fast freezing and slow thawing of Salmonella anatum cells suspended in water resulted in injury of more than 90% of the cells that survived the treatment. The injured cells failed to form colonies on the selective medium (xyloselysine-peptone-agar with 0.2% sodium deoxycholate) but did form colonies on a nonselective (xylose-lysine-peptone-agar) plating medium. In Tryptic soy plus 0.3% yeast extract broth or minimal broth, most of the injured cells repaired within 1 to 2 hr at 25 C. Tryptic soy plus yeast extract broth supported repair to a greater extent than minimal broth. Phosphate or citrate at concentrations found in minimal broth supported repair of some cells. MgSO(4), when present with inorganic phosphate or citrate or both, increased the extent of repair. The repair process in the presence of phosphate was not prevented by actinomycin D, chloramphenicol, and D-cycloserine, but was prevented by cyanide and 2,4-dinitrophenol (only at pH 6). This suggested that the repair process might involve energy metabolism in the form of adenosine triphosphate. The freeze-injured cells were highly sensitive to lysozyme, whereas unfrozen fresh cells were not. In the presence of phosphate or minimal broth this sensitivity was greatly reduced. This suggested that, at least in some of the cells, the injury involved the lipopolysaccharide of the cell wall and adenosine triphosphate synthesis was required for repair.


The role of the C-terminal region in phosphoglycerate mutase.
Removal of the C-terminal seven residues from phosphoglycerate mutase from Saccharomyces cerevisiae by limited proteolysis is associated with loss of mutase activity, but no change in phosphatase activity. The presence of the cofactor 2, 3-bisphosphoglycerate, or of the cofactor and substrate 3-phosphoglycerate together, confers protection against proteolysis. The substrate alone offers no protection. Replacement of either or both of the two lysines at the C-terminus by glycines has only limited effects on the kinetic properties of phosphoglycerate mutase, indicating that these residues are unlikely to be involved in crucial electrostatic interactions with the substrate, intermediate or product in the reaction. However, the double-mutant form of the enzyme is more sensitive to proteolysis and is no longer protected against proteolysis by the presence of cofactor. The proteolysed wild-type and two of the mutated forms of the enzyme show a reduced response to 2-phosphoglycollate, which enhances the instability of the phospho form of the native enzyme. The phosphoglycerate mutase from Schizosaccharomyces pombe, which lacks the analogous C-terminal tail, has an inherently lower mutase activity and is also less responsive to stimulation by 2-phosphoglycollate. It is proposed that the C-terminal region of phosphoglycerate mutase helps to maintain the enzyme in its active phosphorylated form and assists in the retention of the bisphosphoglycerate intermediate at the active site. However, its role seems not to be to contribute directly to ligand binding, but rather to exert indirect effects on the transfer of the phospho group between substrate, enzyme, intermediate and product.


BST2/tetherin inhibits dengue virus release from human hepatoma cells.
Type I interferons (IFN) have been shown to play an important role for inhibiting Dengue virus (DENV) infection. Identifying IFN-induced cellular proteins are essential for understanding its mechanisms against DENV. Here we established stable Huh7-derived cell lines expressing the IFN-induced cell membrane protein BST2 (Huh7-BST2) or its variant bearing a V5 tag at the C-terminal (Huh7-BST5CV5). These cell lines were infected with DENV to determine proteins modulating their anti-DENV response. We found that expression of BST2 did not affect the efficiency of DENV infection and intracellular replication. Rather, it significantly reduced the virion yield of the infected cells, particularly at low MOI infection. In addition, BST2 also decreased the foci formation and the size of infectious foci in cultured Huh7 monolayers with media containing methocellulose. The addition of the V5 tag at C-terminal inhibited the GPI modification of BST2 and blocked its shift from endoplasm to cytoplastic membrane. BST2CV5 did not affect DENV infection and foci formation in Huh7 cells but reduced virion yield by 1 log at low MOI infection. Interestingly, intracellular BST2CV5 expression was reduced by high level of DENV production. Our results imply that BST2 is a functional mediator of the IFN response against DENV infection. BST2 inhibits the release of DENV virions from Huh7 cells and limits viral cell-to-cell transmission. BST2CV5 variant is unable to inhibit DENV release but impairs viral infection in cells.


Flavonoids and phenolic acids from cranberry juice are bioavailable and bioactive in healthy older adults.
Cranberries (Vaccinium macrocarpon) are a rich source of phenolic phytochemicals, which likely contribute to their putative health benefits. A single-dose pharmacokinetic trial was conducted in 10 healthy adults ⩾50y to evaluate the acute (24-h) absorption and excretion of flavonoids, phenolic acids and proanthocyanidins (PACs) from a low-calorie cranberry juice cocktail (54% juice). Inter-individual variability was observed in the Cmax and Tmax of many of these compounds in both plasma and urine. The sum total concentration of phenolics detected in plasma reached a peak of 34.2μg/ml between 8 and 10h, while in urine this peak was 269.8μg/mg creatinine, and appeared 2-4h earlier. The presence of PAC-A2 dimers in human urine has not previously been reported. After cranberry juice consumption, plasma total antioxidant capacity assessed using ORAC and TAP assays correlated with individual metabolites. Our results show phenolic compounds in cranberry juice are bioavailable and exert antioxidant actions in healthy older adults.


Distribution of T-cell markers CD4 and CD8α in lymphoid organs of healthy newborn, juvenile, and adult highland-plateau yaks.
To investigate the distribution of T-cell markers (CD4 and CD8α) in lymphoid organs of newborn, juvenile, and adult yaks. 15 healthy male yaks of various ages from highland plateaus. Yaks were allocated to groups on the basis of age (newborn [1 to 7 days old; n = 5], juvenile [5 to 7 months old; 5], and adult [3 to 4 years old; 5]). The thymus, spleen, 5 mesenteric lymph nodes, and 5 hemal nodes were harvested from each yak within 10 minutes after euthanasia. Morphological characteristics of those lymphoid organs were assessed by histologic examination; expression of CD4 and CD8α mRNAs and proteins were measured by quantitative real-time PCR assay and immunohistochemical staining. Among the lymphoid organs evaluated, expressions of CD4 and CD8α mRNAs were highest in the thymus in all age groups. In newborn lymphoid organs, CD4 mRNA expression and CD4+ cell distribution were more predominant, whereas in juvenile and adult lymphoid organs, CD8α mRNA expression and CD8α+ cell distribution were more predominant. The CD4+ and CD8α+ cells were mainly located in the cortex and medulla of the thymus, the medulla of the hemal nodes and mesenteric lymph nodes, the periarteriolar lymphoid sheaths, and the red pulp of the spleen. Results indicated that the CD4 mRNA expression and CD4+ T-cell distribution in yak lymphoid organs decreased and CD8α mRNA expression and CD8α+ T-cell distribution increased with age. Moreover, CD8α+ cells were present in the follicles of yaks' secondary lymphoid organs, which differs from findings for other mammals.


The effect of 3-(5'-hydroxymethyl-2'-furyl)-1-benzylindazole (YC-1) on cell viability under hypoxia.
The synthetic compound 3-(5'-hydroxymethyl-2'-furyl)-1-benzylindazole (YC-1) reduces the protein stability of hypoxia-inducible factor (HIF)-1α and can serve as a potential anticancer agent. Our previous study elucidated that YC-1 decreased the protein level of HIF-1α and inhibited cell proliferation under normoxic conditions. In the present study, we explored the inhibitory effect of YC-1 on the regulation of HIF-1α and cell survival under hypoxia. Chemical and physical hypoxia using cobalt chloride and an anaerobic incubator, respectively, was induced in the photoreceptor cell line 661W. 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay and morphological observation were used to analyze cell survival. Flow cytometry with a LIVE/DEAD cell viability assay and annexin V was used to determine the number of live and dead cells or cell apoptosis, respectively. Cell proliferation was analyzed with high-content screening of MKI67 (K(i)-67) immunofluorescent staining. Immunoblotting and a quantitative reverse-transcription PCR were used to assess the protein and mRNA levels, respectively. Our results showed that 661W cells exposed to YC-1 decreased cell survival through the induction of cell apoptosis and cell-cycle arrest under hypoxia. We also found that YC-1 reduced the HIF-1α protein level after 2 h of hypoxia, but the mRNA level of HIF-1α was not affected. In addition, YC-1 significantly increased levels of p53, the proapoptotic gene BCL2-associated X protein (Bax), and cell proliferation-related gene, cyclin-dependent kinase inhibitor 1A (p21) mRNAs under hypoxia. Unlike normoxia, YC-1 not only inhibited cell proliferation but also induced cell death under hypoxia. We also found that YC-1 inhibited hypoxia-induced HIF-1α and partially affected hypoxia-regulated gene expression.


(Me2NH2)10[H2-Dodecatungstate] polymorphs: dodecatungstate cages embedded in a variable dimethylammonium cation + water of crystallization matrix.
Two polymorphs and a solvatomorph of a new dimethylammonium polytungstate-decakis(dimethylammonium) dihydrogendodecatungstate, (Me2NH2)10(W12O42)·nH2O (n = 10 or 11)-have been synthesized. Their structures were characterized by single-crystal X-ray diffraction and solid-phase NMR methods. The shape of the dodecatungstate anions is essentially the same in all three structures, their interaction with the cations and water of crystallization, however, is remarkably variable, because the latter forms different hydrogen-bonded networks, and provides a highly versatile matrix. Accordingly, the N-H⋯O and C-H⋯O hydrogen bonds are positioned in each crystal lattice in a variety of environments, characteristic to the structure, which can be distinguished by solid-state 1H-CRAMPS, 13C, 15N CP MAS and 1H-13C heteronuclear correlation NMR. Thermogravimetry of the solvatomorphs also reflect the difference and multiformity of the environment of the water molecules in the different crystal lattices. The major factors behind the variability of the matrix are the ability of ammonium cations to form two hydrogen bonds and the rigidity of the polyoxometalate anion cage. The positions of the oxygen atoms in the latter are favourable for the formation of bifurcated and trifurcated cation-anion hydrogen bonds, some which are so durable that they persist after the crystals are dissolved in water, forming ion associates even in dilute solutions. The H atom involved in furcated hydrogen bonds cannot be exchanged by deuterium when the compound is dissolved in D2O. An obvious consequence of the versatility of the matrix is the propensity of these compounds to form multiple polymorphs.


The challenge of preventing and treating obesity in low-income, preschool children: perceptions of WIC health care professionals.
Obesity has become a common nutritional concern among low-income, preschool children, a primary target population of the Special Supplemental Nutrition Program for Women, Infants, and Children (WIC). Nutrition counseling efforts in WIC target childhood obesity, but new approaches are needed that address the different perceptions about obesity that are held by clients and health care professionals. To develop these new approaches, we examined WIC health care professionals' perceptions about the challenges that exist in preventing and managing childhood obesity. A qualitative study using data transcribed from audiotapes of focus groups and individual interviews. We independently read each transcript and coded themes; then, the common themes were selected through group meetings of the authors. Kentucky WIC. Of the 19 health care professionals participating, all had provided nutrition counseling in WIC and all but one were white women. Twelve major themes clustered into 3 domains. The first domain centered on how WIC health care professionals perceived the life experiences, attitudes, and behaviors of the mothers they counseled. They perceived that mothers (1) were focused on surviving their daily, life stresses; (2) used food to cope with these stresses and as a tool in parenting; (3) had difficulty setting limits with their children around food; (4) lacked knowledge about normal child development and eating behavior; (5) were not committed to sustained behavioral change; and (6) did not believe their overweight children were overweight. The second domain described WIC health care professionals' perceptions of counseling interactions. They felt that (7) they might offend mothers when talking about weight, (8) counseling was driven by protocols, and (9) their nutritional advice often conflicted with the advice from the mothers' relatives, friends, or primary care physicians. The last domain described programmatic suggestions WIC health care professionals offered to address childhood obesity: These included (10) promoting a more client-centered approach to counseling, (11) establishing behavioral change goals that were small and endorsed by the mother, and (12) working with primary care physicians to create a more uniform approach to counseling on obesity. To become more responsive to the problem of childhood obesity, WIC should consider the following: (1) providing staff training in counseling skills that educate parents on child development and child-rearing and that elicit the client's social context and personal goals, (2) shifting time allocation and programmatic emphasis in the WIC visits away from nutritional risk assessment and toward counseling, and (3) developing collaborations with primary health care providers and community agencies that impact childhood obesity.


Impact of the Council of Europe Resolution on quality and safety assurance requirements for medicinal products prepared in pharmacies for the special needs of patients.
The regulation of pharmacy preparations, especially for standards for quality assurance and safety, is not harmonised across Europe and falls under the national competencies of individual states. There are concerns about quality control and safety for the medicinal products made in pharmacies, which is widespread in European countries. There are, however, good reasons to continue this practice, which is able to tailor preparations to the specific needs of a particular patient or patient group and to provide a supplementary source of supply when an industrially manufactured product, which is authorised for marketing is not available or when there are temporary shortages of licensed medicines. In seeking to provide guidelines for legislation and acting on the advice of an expert group dealing in pharmaceutical practices, the Committee of Ministers of the Council of Europe passed a resolution in 2011. The Council of Europe Resolution provides authorities and pharmacists with the means to reinforce safety measures for medicinal products prepared in pharmacies and to harmonise quality assurance and safety standards. It dealt with aspects of pharmacy preparation such as quality standards for preparation and distribution, marketing authorisation, product dossiers, labelling, reporting, and safety. In 2013 and 2014 the Committee of Experts carried out a survey to evaluate the impact of the resolution within a cross section of member states. The objectives of this study were both to monitor the extent to which the recommendations had been enshrined in national legislation and also to understand current differences in legislation and practice between the member states. In the resolution of 2011 the member states were recommended to adapt their legislation in line with its provisions. The survey that was carried out in 2013 and 2014 followed the recommendations in the resolution. A questionnaire was made and sent to a cross section of member states. Among the member states involved, the results of this survey show a clear commitment to implement the recommendations of the resolution. This report presents the results of the survey with a discussion of outstanding issues.


'It could have been a lot worse': the psychological effects of farm-related serious injury in Victoria.
The psychological response to injury is an important factor in recovery, and the development of psychological problems can result in a delay or inability to return to pre-injury function, including return to work. Farming is widely acknowledged as a stressful occupation, with non-injured farmers already at an increased risk of developing significant psychological problems, including high levels of stress and depression, and increased rates of suicide. This study aimed to investigate the psychological effects of serious farm-related injury on farmers, and how this influences their recovery. This was a qualitative study of 31 farmers in the state of Victoria, Australia, who sustained major trauma between 2007 and 2013. Participants were identified using the Victorian State Trauma Registry and underwent an in-depth, semi-structured telephone interview. Recruitment continued until data saturation was achieved, and thematic analysis was used to identify important themes from the data. For many farmers, the traumatic circumstances and ongoing impact of their injury are life-changing. In this study, the psychological effects of sustaining a major farm-related injury varied between participants; however, four major interconnected themes were identified: importance of a pragmatic outlook; grief, helplessness and loss of independence; traumatic thoughts post-injury; and the importance of the support network and community. The findings of this study highlight both a reluctance for psychological assistance as well as the importance of psychological resilience and support networks to recovery. Overall, a positive outlook was found to be the primary enabler in the farmers' recovery. It is recommended that injured farmers should be provided with additional psychological support and advice to aid in their recovery. Additionally, psychological support services should be extended to include both the family network and the broader farming community, as these were found to experience significant short- and long-term stress following farmers' injury.


Cesena guidelines: WSES consensus statement on laparoscopic-first approach to general surgery emergencies and abdominal trauma.
Laparoscopy is widely adopted across nearly all surgical subspecialties in the elective setting. Initially finding indication in minor abdominal emergencies, it has gradually become the standard approach in the majority of elective general surgery procedures. Despite many technological advances and increasing acceptance, the laparoscopic approach remains underutilized in emergency general surgery and in abdominal trauma. Emergency laparotomy continues to carry a high morbidity and mortality. In recent years, there has been a growing interest from emergency and trauma surgeons in adopting minimally invasive surgery approaches in the acute surgical setting. The present position paper, supported by the World Society of Emergency Surgery (WSES), aims to provide a review of the literature to reach a consensus on the indications and benefits of a laparoscopic-first approach in patients requiring emergency abdominal surgery for general surgery emergencies or abdominal trauma. This position paper was developed according to the WSES methodology. A steering committee performed the literature review and drafted the position paper. An international panel of 54 experts then critically revised the manuscript and discussed it in detail, to develop a consensus on a position statement. A total of 323 studies (systematic review and meta-analysis, randomized clinical trial, retrospective comparative cohort studies, case series) have been selected from an initial pool of 7409 studies. Evidence demonstrates several benefits of the laparoscopic approach in stable patients undergoing emergency abdominal surgery for general surgical emergencies or abdominal trauma. The selection of a stable patient seems to be of paramount importance for a safe adoption of a laparoscopic approach. In hemodynamically stable patients, the laparoscopic approach was found to be safe, feasible and effective as a therapeutic tool or helpful to identify further management steps and needs, resulting in improved outcomes, regardless of conversion. Appropriate patient selection, surgeon experience and rigorous minimally invasive surgical training, remain crucial factors to increase the adoption of laparoscopy in emergency general surgery and abdominal trauma. The WSES expert panel suggests laparoscopy as the first approach for stable patients undergoing emergency abdominal surgery for general surgery emergencies and abdominal trauma.


Breast Asymmetry in Women Requesting Plastic Surgery of the Breast.
Breast asymmetries have a high prevalence among women requesting breast augmentation. However, the prevalence of breast asymmetries in women undergoing other types of breast surgeries is not known. The breast measurements of consecutively women evaluated for plastic surgery of the breast, but without prior breast surgery, were prospectively recorded in a plastic surgery database. They were classified into three groups according to the presenting breast problem: hypoplastic breasts, macromastia, and ptotic breasts. Comparisons were made between the right and left side of each patient regarding the symmetry of the nipple-areola complex (size and position), breast mound, and chest wall. The breast measurements of 304 women were analyzed. The mean age was 35 ± 12 years. The study population was distributed in the following manner: 126 hypoplastic breast cases, 100 macromastia cases, and 78 ptotic breast cases. Asymmetry of the position of the nipple-areola complex was found in 54%, 59%, and 56% of the groups, respectively. Asymmetry of the breast mound volume was found in 41%, 47%, and 44% of the groups. Asymmetry of the chest wall was present in 12%, 11%, and 10% of the groups, respectively. Overall, we found that 91% of the cases had at least one type of breast asymmetry. Prevalence of asymmetry was not different (p>0.05) among the groups, but the magnitude was larger in macromastia. Breast asymmetries were detected in the majority of women and the prevalence was similar across the different groups, however the magnitude was greater in hypertrophic breasts.


Electron transfer dynamics from single near infrared emitting lead sulfide-cadmium sulfide nanocrystals to titanium dioxide.
In this study we report the first successful demonstration of electron transfer between single near infrared emitting PbS/CdS nanocrystals and an external acceptor, titanium dioxide (TiO2). We demonstrate distance-dependent electron transfer from single nanocrystals to TiO2 and explore the effect of this process on the photoluminescence dynamics of these nanocrystals. Isolated PbS/CdS QDs are found to exhibit blinking dynamics similar to other nanocrystals like CdSe/ZnS; however, their photoluminescence follows a quasi two-state pattern with heterogeneous photoluminescence lifetimes which may be the result of their emission originating from different energy states. Electron transfer of these nanocrystals with an external acceptor inhibits their photoluminescence lifetime heterogeneity and biases their blinking dynamics in a manner similar to that observed for visible emitting CdSe/ZnS nanocrystals undergoing electron transfer with external acceptors. While the present study reconfirms the universality of quantum dot blinking among various types of nanocrystals, it also demonstrates that universality remains valid for the communication of various types of nanocrystals with the exterior world, here pictured as electron transfer with external acceptors.


Dietary and genetic influences on hemostasis in a Yup'ik Alaska Native population.
Fish and marine animals are important components of the subsistence diet of Alaska Native people, resulting in a high ω3 PUFA intake. The historical record for circumpolar populations highlights a tendency for facile bleeding, possibly related to ω3 PUFA effects on platelet activation and/or vitamin K-dependent clotting factors. To evaluate these two scenarios in Yup'ik people of southwestern Alaska, we examined the association between dietary ω3 PUFA intake and activities of clotting factor II, V, fibrinogen, PT, INR, PTT, and sP-selectin in 733 study participants, using the nitrogen isotope ratio of red blood cells as a biomarker of ω3 PUFA consumption. sP-selectin alone correlated strongly and inversely with ω3 PUFA consumption. Approximately 36% of study participants exhibited PIVKA-II values above the threshold of 2 ng/ml, indicative of low vitamin K status. To assess genetic influences on vitamin K status, study participants were genotyped for common vitamin K cycle polymorphisms in VKORC1, GGCX and CYP4F2. Only CYP4F2*3 associated significantly with vitamin K status, for both acute (plasma vitamin K) and long-term (PIVKA-II) measures. These findings suggest: (i) a primary association of ω3 PUFAs on platelet activation, as opposed to vitamin K-dependent clotting factor activity, (ii) that reduced CYP4F2 enzyme activity associates with vitamin K status. We conclude that high ω3 PUFA intake promotes an anti-platelet effect and speculate that the high frequency of the CYP4F2*3 allele in Yup'ik people (~45%) evolved in response to a need to conserve body stores of vitamin K due to environmental limitations on its availability.


Decline in physical functioning in first 2 years after breast cancer diagnosis predicts 10-year survival in older women.
Breast cancer patients often experience a decline in physical functioning following cancer diagnosis. Although most patients recover after treatment, some patients do not. These changes may be magnified in older women with comorbid conditions and could impact survival outcomes. We used longitudinal data from a prospective cohort study of women 65+ years of age, recruited shortly after diagnosis of early stage breast cancer, to examine changes in self-reported physical functioning measured with the Physical Function Index (PF-10) of the Medical Outcomes Study Short Form-36. Outcomes were constructed for small (0.2 SD), medium (0.5 SD), and large (0.8 SD) declines in the PF-10 measurement over two intervals: (1) 3 to 15 months following cancer diagnosis, encompassing treatment and early recovery, and (2) 3 to 27 months following cancer diagnosis, in order to detect sustained recovery versus persistent decline. Cox-proportional hazards regression was used to examine association between survival and decline in PF-10 scores. A large (>0.8 SD) decline in PF-10 scores from 3 to 27 months predicted shorter 10-year survival (hazard ratio = 1.34, 95 % confidence interval 1.1-1.6). Persistent decline at 27 months was associated with less education, higher baseline PF-10, increased comorbidity, and higher body mass index. Older women with breast cancer who experience a large and persistent decline in PF-10 are at increased mortality risk. Future research should examine the value of clinical assessment of physical function as a marker for mortality and test interventions to prevent decline in physical function to improve post-treatment survival outcomes. Many breast cancer survivors experience a decline in physical functioning in the year following their cancer diagnosis. While the majority of patients recover in the year after treatment is completed, individuals who experience a persistent decline in physical functioning without recovery are at risk of functional decline and early mortality. Our results highlight the importance of assessing functional status to identify those at risk for functional decline, and suggest a potential role for early intervention in these individuals.


Platelet-bound ristocetin aggregation factor in normal subjects and patients with von Willebrand's disease.
Antiserum specific for that part of the factor VIII complex designated ristocetin aggregation factor (VIIIRAF) was prepared by immunizing rabbits with VIIIRAF derived from the plasma of a hemophilic patient with circulating antibody to factor VIII procoagulant activity (VIIIcoag). The rabbit antiserum prevented ristocetin-induced platelet aggregation and platelet retention by glass-bead columns. Although the antiserum also inactivated VIIIcoag in normal plasma, it did not inactivate VIIIcoag which had been dissociated from VIIIRAF by chromatography in 1 M NaCl, thus establishing the antigenic specificity of these two factors. When the VIIIRAF antibody was conjugated with fluorescein isothiocyanate and used to examine platelets from normal subjects and patients with von Willebrand's disease, the normal platelets showed a granular fluorescence similar to that observed with antifibrinogen serum whole von Willebrand platelets showed no fluorescent staining. The normal platelets retained the VIIIRAF granules during 18 hours incubation and 5 washings in artificial medium while the von Willebrand platelets failed to acquire granules after 18 hours in normal plasma. When the unstained platelets from patients with von Willebrand's disease were suspended in normal plasma and then aggregated by the addition of ristocetin, the aggregates not only stained brightly for VIIRAF, but fluorescent granules could be seen on individual platelets in the aggregates. These observations suggest that ristocetin causes the binding of VIIIRAF to platelets as well as platelet-to-platelet adhesion.


Deletion of membrane-associated Asp23 leads to upregulation of cell wall stress genes in Staphylococcus aureus.
With about 25 000 molecules per cell, Asp23 is one of the most abundant proteins in Staphylococcus aureus. Asp23 has been characterized as a protein that, following an alkaline shock, accumulates in the soluble protein fraction. Transcription of the asp23 gene is exclusively regulated by the alternative sigma factor σ(B) , which controls the response of the bacterium to environmental stress. Sequence analysis identified Asp23 as a member of the widely distributed Pfam DUF322 family, precluding functional predictions based on its sequence. Using fluorescence microscopy we found that Asp23 colocalized with the cell membrane of Staphylococcus aureus. Since Asp23 has no recognizable transmembrane spanning domains, we initiated a search for proteins that link Asp23 to the cell membrane. We identified SAOUHSC_02443 as the Asp23 membrane anchor and have renamed it AmaP (Asp23 membrane anchoring protein). Deletion of the asp23 gene led to an upregulation of the cell wall stress response. In summary, we have identified Asp23 as a membrane-associated protein and we suggest a function for Asp23 in cell envelope homoeostasis.


Heterogeneity of the composition and thickness of tracheal mucus in rats.
Inability to preserve airway mucus in situ has limited our understanding of its structure and function. This light- and transmission electron-microscopic study of rat tracheal mucus used a nonaqueous fixative that retains mucus (epiphase) over a lucent layer (hypophase). The fixative is a 1% solution of osmium tetroxide dissolved in a perfluorocarbon. The mean thickness of rat tracheal epiphase was 5 microns, with significant variation (0.1-50 microns) around the tracheal circumference. Tracheal mucus was thickest at the trachealis muscle region and contained cells, cellular debris, and a variable amount of surfactant and lipid, estimated at 4-16% of the total epiphase in five rats, with a mean composition of 9%. Lipid was observed on the surface of the epiphase, embedded within mucus, and at the epiphase-hypophase interface. Refined study of developmental, physiological, and pathological alterations to the airway coat may benefit from this approach.


Kinetic and computational evidence for an intermediate in the hydrolysis of sulfonate esters.
The hydrolytic reactions of sulfonate esters have previously been considered to occur by concerted mechanisms. We now report the observation of a break in a Brønsted correlation for the alkaline hydrolysis of aryl benzenesulfonates. On either side of a break-point, β(leaving group) values of -0.27 (pK(a) < 8.5) and -0.97 (pK(a) > 8.5) are measured. These data are consistent with a two-step mechanism involving a pentavalent intermediate that is also supported by QM/MM calculations. The emerging scenario can be explained by the combined effect of a strong nucleophile with a poor leaving group that compel a usually concerted reaction to favour a stepwise process.


Association of argyrophilic nucleolar organizing regions, Ki-67, and proliferating cell nuclear antigen scores with histologic grade and survival in dogs with soft tissue sarcomas: 60 cases (1996-2002).
To determine whether argyrophilic nucleolar organizing regions (AgNORs), Ki-67, and proliferating cell nuclear antigen (PCNA) scores were associated with histologic grade and survival in dogs with soft tissue sarcomas (STSs). Retrospective study. 60 dogs with STSs. Medical records were examined and histologic specimens were reviewed. Tissue specimens obtained from archival materials were used to prepare sections for histologic staining for AgNOR and immunohistochemical staining for Ki-67 and PCNA labeling. Follow-up monitoring was obtained by reevaluation or telephone conversations with referring veterinarians or owners. 27 (45%) STSs were grade 1, 23 (38%) were grade 2, and 10 (17%) were grade 3. The mean and median AgNOR, Ki-67, and PCNA scores were determined, and significant positive associations among AgNOR and Ki-67 scores with histologic grade and mitotic score were detected. Fifty-four dogs had adequate follow-up examinations and were included in survival analysis and evaluation of prognostic factors. Overall median survival time was > 1,306 days. Twelve of 54 (22%) dogs died of tumor-related causes. Metastatic disease developed in 8 of 54 (15%) dogs. Results of univariate analysis indicated that increased mitotic score, increased AgNOR score, increased Ki-67 score, incomplete surgical margins, noncurative intent surgery, Ki-67 score greater than the median Ki-67 score, and AgNOR score greater than the median AgNOR score were prognostic factors for decreased survival time. Results of multivariate analysis indicated that increased AgNOR score was the only prognostic factor for decreased survival time. Results suggested that AgNORs and possibly Ki-67 should be routinely evaluated with histologic grading for STSs in dogs.


Implications of HLA sequence-based typing in transplantation.
Serology-based conventional microlymphocytotoxicity HLA typing method, which has been regarded as the gold standard in organ and hematopoietic stem cell transplantation, has been replaced now by DNA-based typing. Many laboratories all over the world have already switched over to molecular methods. Microlymphocytotoxicity-based tissue typing was done using commercial sera, while the molecular typing by genomic DNA based. DNA quality and its quantity obtained using various DNA extraction protocols was found to be an important factor in the molecular method of tissue typing in transplant outcome. Many polymerase chain reaction-based molecular techniques have been adopted with far reaching clinical outcome. The sequence-based typing (SBT) has been the ultimate technique, which has been of the highest reliability in defining the HLA alleles. The nonavailability of specific HLA antisera from native populations, large number of blank alleles yet to be defined and comparable low resolution of HLA alleles in SSP or SSOP technique, suggests that highly refined DNA-based methods like SBT should be used as an adjunct to HLA serology and/or low/intermediate/high resolution HLA typing in order to achieve a better transplant outcome.


Nitric oxide stress in sporadic inclusion body myositis muscle fibres: inhibition of inducible nitric oxide synthase prevents interleukin-1β-induced accumulation of β-amyloid and cell death.
Sporadic inclusion body myositis is a severely disabling myopathy. The design of effective treatment strategies is hampered by insufficient understanding of the complex disease pathology. Particularly, the nature of interrelationships between inflammatory and degenerative pathomechanisms in sporadic inclusion body myositis has remained elusive. In Alzheimer's dementia, accumulation of β-amyloid has been shown to be associated with upregulation of nitric oxide. Using quantitative polymerase chain reaction, an overexpression of inducible nitric oxide synthase was observed in five out of ten patients with sporadic inclusion body myositis, two of eleven with dermatomyositis, three of eight with polymyositis, two of nine with muscular dystrophy and two of ten non-myopathic controls. Immunohistochemistry confirmed protein expression of inducible nitric oxide synthase and demonstrated intracellular nitration of tyrosine, an indicator for intra-fibre production of nitric oxide, in sporadic inclusion body myositis muscle samples, but much less in dermatomyositis or polymyositis, hardly in dystrophic muscle and not in non-myopathic controls. Using fluorescent double-labelling immunohistochemistry, a significant co-localization was observed in sporadic inclusion body myositis muscle between β-amyloid, thioflavine-S and nitrotyrosine. In primary cultures of human myotubes and in myoblasts, exposure to interleukin-1β in combination with interferon-γ induced a robust upregulation of inducible nitric oxide synthase messenger RNA. Using fluorescent detectors of reactive oxygen species and nitric oxide, dichlorofluorescein and diaminofluorescein, respectively, flow cytometry revealed that interleukin-1β combined with interferon-γ induced intracellular production of nitric oxide, which was associated with necrotic cell death in muscle cells. Intracellular nitration of tyrosine was noted, which partly co-localized with amyloid precursor protein, but not with desmin. Pharmacological inhibition of inducible nitric oxide synthase by 1400W reduced intracellular production of nitric oxide and prevented accumulation of β-amyloid, nitration of tyrosine as well as cell death inflicted by interleukin-1β combined with interferon-γ. Collectively, these data suggest that, in skeletal muscle, inducible nitric oxide synthase is a central component of interactions between interleukin-1β and β-amyloid, two of the most relevant molecules in sporadic inclusion body myositis. The data further our understanding of the pathology of sporadic inclusion body myositis and may point to novel treatment strategies.


Conductance and kinetics of delayed rectifier potassium channels in nodal cells of the rabbit heart.
1. The delayed rectifier K+ current (IK) of single pace-maker cells from the sino-atrial node and the atrioventricular node of the rabbit heart was investigated using the whole-cell and cell-attached configurations of the patch-clamp technique. 2. The activation kinetics of the macroscopic IK were not altered by varying the extracellular K+ concentration ([K+]o) between 5.4 and 150 mM. The amplitude of the tail current of IK, however, was about 10-fold larger at a [K+]o of 150 mM than that at a [K+]o of 5.4 mM. 3. By using a high-[K+]o solution, inward single-channel currents were observed on repolarization from potentials positive to -40 mV. The current-voltage (I-V) relation was linear over the negative potential range and the reversal potential estimated by extrapolating the I-V curve was shifted by about 60 mV for a 10-fold increase in [K+]o, indicating that the channel was highly selective for K+. 4. The single-channel conductance was 11.1 pS at a [K+]o of 150 mM and varied in proportion to the square root of [K+]o. The total number of channels was estimated as approximately 1000 per cell (0.7/micron 2). On repolarization, the averaged single-channel current disappeared with a time constant similar to that of the macroscopic tail current of IK. 5. At potentials between -50 and -100 mV, the open and closed times of the single channel fitted well with single-exponential and biexponential distributions, respectively. As the membrane was progressively depolarized, the open time was shortened while the closed time was prolonged, suggesting a decrease of open probability. These changes were in the opposite direction to those expected from the delayed rectifier K+ current which progressively increases in magnitude at more positive potentials. 6. At the beginning of the macroscopic tail current, a transient increase of the inward current was found to precede the time-dependent decrease. This rapid initial change can be attributed to a quick removal of inactivation of IK which had occurred during the depolarizing pulse. This inactivation gate of the channel has very fast kinetics and could be responsible for the inward-going rectification observed in the 'fully activated' IK.


In vitro characterization of the cytokine profile of the epithelial cell rests of Malassez.
The epithelial cell rests of Malassez (ERM) are an integral part of the periodontal ligament and are considered to play an important role in dental pathology. Surprisingly, this cell type is poorly described and is often disregarded in the context of periodontal research. The aim of this study was to establish primary cell cultures of human ERM, characterize the cytokine profile, and compare it to other periodontal cell entities. ERM-derived epithelial cells were isolated from the periodontal ligament of three subjects. A cytokine antibody array, including 120 cytokines in two membranes, was used to determine the cytokine profile of conditioned medium from the ERM-derived epithelial cells. The results were compared to those of gingival epithelial cells and periodontal ligament fibroblasts. ERM-derived epithelial cells expressed 29 of 120 cytokines in significant amounts, including cytokines, chemokines, growth factors, and related proteins, such as interleukin (IL)-1, -6, -8, and -10; granulocyte macrophage-colony stimulating factor; monocyte chemoattractant protein (MCP)-1, -2, and -3; amphiregulin; glial-derived neurotrophic factor; vascular endothelial growth factor; and insulin-like growth factor binding protein-2. The cytokine profile of ERM cells was similar to that of gingival epithelial cells but strikingly different from the profile of periodontal ligament fibroblasts. The results indicated that, via paracrine secretion of a variety of soluble factors, the ERM cells actively take part in the homeostasis of the periodontium. Therefore, future research on the pathophysiology of periodontal tissue should include this often overlooked cell type.


Estrogen-induced epigenetic silencing of FTH1 and TFRC genes reduces liver cancer cell growth and survival.
Estrogen (E2) regulates hundreds of genes involved in cell metabolism and disrupts iron homoeostasis in various cell types. Herein, we addressed whether E2-induced epigenetic modifications are involved in modulating the expression of iron-regulatory genes. Epigenetic status of FTH1 and TFRC genes was assessed in E2-treated cancer cells. E2-induced DNA methylation was associated with decreased FTH1 and TFRC expression in Hep-G2 and Huh7 cells, but not in AGS or MCF7 cells. Demethylation with 5-Aza-2-deoxycytidine upregulated the expression of both these genes in Hep-G2 cells. The expression of DNMT3B, PRMT5, and H4R3me2s increased in E2-treated cells. Chromatin immunoprecipitation showed that E2 treatment recruited PRMT5 and H4R3me2s on FTH1 but not on TFRC. Knockdown of PRMT5, DNMT3B, and Estrogen-receptor alpha rescued FTH1 from E2-induced silencing. However, knockdown of DNMT3B alone blocked the inhibitory effects of E2 on TFRC. Analysis of human liver tissues in publicly available datasets showed that FTH1 and TFRC are highly expressed in primary liver tumours, but a lower expression is associated with better survival. Interestingly, we showed that the silencing of FTH1 and/or TFRC inhibited carcinogenesis in Hep-G2 cells. For the first time, our findings uncovered the novel signalling pathway involved in the protective effects of E2 against liver cancer.


Changes in mitochondrial membrane composition and oxidative status during rapid growth, maturation and aging in zebrafish, Danio rerio.
Considering membranes and membrane components as possible pacemakers of the main processes taking place inside mitochondria, changes in phospholipids or fatty acids could play a central role linking different mechanisms involved in cumulative damage to cell molecules and dysfunction during periods of high stress, such as rapid growth and aging. Changes affecting either lipid class or fatty acid compositions could affect phospholipid and membrane properties and alter mitochondrial function and cell viability. In the present study, mitochondrial oxidative status and mitochondrial membrane phospholipid compositions were analyzed throughout the life-cycle of zebrafish. TBARS content significantly increased in 18-month-old fish while aconitase activity decreased in 24-month-old fish, which have been related with oxidative damage to molecules. Mitochondria-specific superoxide dismutase decreased in 24-month-old animals although this change was not statistically significant. Age affected both mitochondrial phospholipid content and the peroxidation index of most phospholipid classes suggesting that oxidative damage to mitochondrial lipids was occurring.


Monolayer WS2 Nanopores for DNA Translocation with Light-Adjustable Sizes.
Two-dimensional materials are promising for a range of applications, as well as testbeds for probing the physics of low-dimensional systems. Tungsten disulfide (WS2) monolayers exhibit a direct band gap and strong photoluminescence (PL) in the visible range, opening possibilities for advanced optoelectronic applications. Here, we report the realization of two-dimensional nanometer-size pores in suspended monolayer WS2 membranes, allowing for electrical and optical response in ionic current measurements. A focused electron beam was used to fabricate nanopores in WS2 membranes suspended on silicon-based chips and characterized using PL spectroscopy and aberration-corrected high-resolution scanning transmission electron microscopy. It was observed that the PL intensity of suspended WS2 monolayers is ∼10-15 times stronger when compared to that of substrate-supported monolayers, and low-dose scanning transmission electron microscope viewing and drilling preserves the PL signal of WS2 around the pore. We establish that such nanopores allow ionic conductance and DNA translocations. We also demonstrate that under low-power laser illumination in solution, WS2 nanopores grow slowly in size at an effective rate of ∼0.2-0.4 nm/s, thus allowing for atomically controlled nanopore size using short light pulses.


The effect of low temperature aging on the mechanical property & phase stability of Y-TZP ceramics.
Recently Yttrium-stabilized tetragonal zirconia polycrystal (Y-TZP) has been introduced due to superior flexural strength and fracture toughness compared to other dental ceramic systems. Although zirconia has outstanding mechanical properties, the phenomenon of decrease in the life-time of zirconia resulted from degradation in flexural strength after low temperature aging has been reported. The objective of this study was to investigate degradation of flexural strength of Y-TZP ceramics after various low temperature aging treatments and to evaluate the phase stability and micro-structural change after aging by using X-ray diffraction analysis and a scanning electron microscope (SEM). Y-TZP blocks of Vita In-Ceram YZ (Vita Zahnfabrik, Bad Säckingen, Germany) were prepared in 40 mm (length) × 4 mm (width) × 3 mm (height) samples. Specimens were artificially aged in distilled water by heat-treatment at a temperature of 75, 100, 125, 150, 175, 200, and 225℃ for 10 hours, in order to induce the phase transformation at the surface. To measure the mechanical property, the specimens were subjected to a four-point bending test using a universal testing machine (Instron model 3365; Instron, Canton, Mass, USA). In addition, X-ray diffraction analysis (DMAX 2500; Rigaku, Tokyo, Japan) and SEM (Hitachi s4700; Jeol Ltd, Tokyo, Japan) were performed to estimate the phase transformation. The statistical analysis was done using SAS 9.1.3 (SAS institute, USA). The flexural strength data of the experimental groups were analyzed by one-way analysis of variance and to detect statistically significant differences (α= .05). The mean flexural strength of sintered Vita In-Ceram YZ without autoclaving was 798 MPa. When applied aging temperature at below 125℃ for 10 hours, the flexural strength of Vita In-Ceram YZ increased up to 1,161 MPa. However, at above 150℃, the flexural strength started to decrease. Although low temperature aging caused the tetragonal-to-monoclinic phase transformation related to temperature, the minimum flexural strength was above 700 MPa. The monoclinic phase started to appear after aging treatment above 100℃. With the higher aging temperature, the fraction of monoclinic phase increased. The ratio of monoclinic/tetragonal + monoclinic phase reached a plateau value, circa 75% above 175℃. The point of monoclinic concentration at which the flexural strength begins to decrease was between 12% and 54%.


Biomedical Applications of Antiviral Nanohybrid Materials Relating to the COVID-19 Pandemic and Other Viral Crises.
The COVID-19 pandemic has driven a global research to uncover novel, effective therapeutical and diagnosis approaches. In addition, control of spread of infection has been targeted through development of preventive tools and measures. In this regard, nanomaterials, particularly, those combining two or even several constituting materials possessing dissimilar physicochemical (or even biological) properties, i.e., nanohybrid materials play a significant role. Nanoparticulate nanohybrids have gained a widespread reputation for prevention of viral crises, thanks to their promising antimicrobial properties as well as their potential to act as a carrier for vaccines. On the other hand, they can perform well as a photo-driven killer for viruses when they release reactive oxygen species (ROS) or photothermally damage the virus membrane. The nanofibers can also play a crucial protective role when integrated into face masks and personal protective equipment, particularly as hybridized with antiviral nanoparticles. In this draft, we review the antiviral nanohybrids that could potentially be applied to control, diagnose, and treat the consequences of COVID-19 pandemic. Considering the short age of this health problem, trivially the relevant technologies are not that many and are handful. Therefore, still progressing, older technologies with antiviral potential are also included and discussed. To conclude, nanohybrid nanomaterials with their high engineering potential and ability to inactivate pathogens including viruses will contribute decisively to the future of nanomedicine tackling the current and future pandemics.


Gastric banding or bypass? A systematic review comparing the two most popular bariatric procedures.
Bariatric surgical procedures have increased exponentially in the United States. Laparoscopic adjustable gastric banding is now promoted as a safer, potentially reversible and effective alternative to Roux-en-Y gastric bypass, the current standard of care. This study evaluated the balance of patient-oriented clinical outcomes for laparoscopic adjustable gastric banding and Roux-en-Y gastric bypass. The MEDLINE database (1966 to January 2007), Cochrane clinical trials database, Cochrane reviews database, and Database of Abstracts of Reviews of Effects were searched using the key terms gastroplasty, gastric bypass, laparoscopy, Swedish band, and gastric banding. Studies with at least 1 year of follow-up that directly compared laparoscopic adjustable gastric banding with Roux-en-Y gastric bypass were included. Resolution of obesity-related comorbidities, percentage of excess body weight loss, quality of life, perioperative complications, and long-term adverse events were the abstracted outcomes. The search identified 14 comparative studies (1 randomized trial). Few studies reported outcomes beyond 1 year. Excess body weight loss at 1 year was consistently greater for Roux-en-Y gastric bypass than laparoscopic adjustable gastric banding (median difference, 26%; range, 19%-34%; P < .001). Resolution of comorbidities was greater after Roux-en-Y gastric bypass. In the highest-quality study, excess body weight loss was 76% with Roux-en-Y gastric bypass versus 48% with laparoscopic adjustable gastric banding, and diabetes resolved in 78% versus 50% of cases, respectively. Both operating room time and length of hospitalization were shorter for those undergoing laparoscopic adjustable gastric banding. Adverse events were inconsistently reported. Operative mortality was less than 0.5% for both procedures. Perioperative complications were more common with Roux-en-Y gastric bypass (9% vs 5%), whereas long-term reoperation rates were lower after Roux-en-Y gastric bypass (16% vs 24%). Patient satisfaction favored Roux-en-Y gastric bypass (P=.006). Weight loss outcomes strongly favored Roux-en-Y gastric bypass over laparoscopic adjustable gastric banding. Patients treated with laparoscopic adjustable gastric banding had lower short-term morbidity than those treated with Roux-en-Y gastric bypass, but reoperation rates were higher among patients who received laparoscopic adjustable gastric banding. Gastric bypass should remain the primary bariatric procedure used to treat obesity in the United States.


Fermionic State Discrimination by Local Operations and Classical Communication.
We consider the problem of local operations and classical communication (LOCC) discrimination between two bipartite pure states of fermionic systems. We show that, contrary to the case of quantum systems, for fermionic systems it is generally not possible to achieve the ideal state discrimination performances through LOCC measurements. On the other hand, we show that an ancillary system made of two fermionic modes in a maximally entangled state is a sufficient additional resource to attain the ideal performances via LOCC measurements. The stability of the ideal results is studied when the probability of preparation of the two states is perturbed, and a tight bound on the discrimination error is derived.


Rare complications of osteolysis and periprosthetic tissue reactions after hybrid and non-hybrid total disc replacement.
Few complications have been reported for lumbar total disc replacement (TDR) and hybrid TDR fixations. This study evaluated retrieved implants and periprosthetic tissue reactions for two cases of osteolysis following disc arthroplasty with ProDisc-L prostheses. Implants were examined for wear and surface damage, and tissues for inflammation, polyethylene wear debris (polarized light microscopy) and metal debris (energy-dispersive X-ray spectroscopy). Despite initial good surgical outcomes, osteolytic cysts were noted in both patients at vertebrae adjacent to the implants. For the hybrid TDR case, heterotopic ossification and tissue necrosis due to wear-induced inflammation were observed. In contrast, the non-hybrid implant showed signs of abrasion and impingement, and inflammation was observed in tissue regions with metal and polyethylene wear debris. In both cases, wear debris and inflammation may have contributed to osteolysis. Surgeons using ProDisc prostheses should be aware of these rare complications.


Sympathetic nervous system activity is associated with obesity-induced subclinical organ damage in young adults.
Excess weight is established as a major risk factor for cardiovascular diseases, particularly in young individuals. To get a better understanding of the pathophysiology underlying increased cardiovascular disease risk, we evaluated early signs of organ damage and their possible relationship to sympathetic nervous activity. Eighteen lean (body mass index <25 kg/m(2)) and 25 overweight or obese (body mass index >25 kg/m(2)) healthy university students were included in the study. We comprehensively assessed subclinical target organ damage, including the following: (1) assessment of renal function; (2) left ventricular structure and systolic and diastolic function; and (3) endothelial function. Muscle sympathetic nervous activity was assessed by microneurography. Participants with excess weight had decreased endothelial function (P<0.01), elevated creatinine clearance (P<0.05), increased left ventricular mass index (P<0.05), increased left ventricular wall thickness (P<0.01), lower systolic and diastolic function (P<0.01), and elevated muscle sympathetic nervous activity (P<0.001) compared with lean individuals. In multiple regression analysis, endothelial function was inversely related to muscle sympathetic nervous activity (R(2)=0.244; P<0.05), whereas creatinine clearance and left ventricular mass index were positively related to muscle sympathetic nervous activity, after adjustment for body mass index, sex, and blood pressure (R(2)=0.318, P<0.01 and R(2)=0.312, P<0.05, respectively). Excess weight in young individuals is associated with subclinical alterations in renal and endothelial function, as well as in the structure of the heart, even in the absence of hypertension. Sympathetic activity is closely associated with cardiovascular and renal alterations observed in these subjects.


Profiling Anti-Neu5Gc IgG in Human Sera with a Sialoglycan Microarray Assay.
Cells are covered with a cloak of carbohydrate chains (glycans) that is commonly altered in cancer and that includes variations in sialic acid (Sia) expression. These are acidic sugars that have a 9-carbon backbone and that cap vertebrate glycans on cell surfaces. Two of the major Sia forms in mammals are N-acetylneuraminic acid (Neu5Ac) and its hydroxylated form, N-glycolylneuraminic acid (Neu5Gc). Humans cannot produce endogenous Neu5Gc due to the inactivation of the gene encoding cytidine 5'monophosphate-Neu5Ac (CMP-Neu5Ac) hydroxylase (CMAH). Foreign Neu5Gc is acquired by human cells through the dietary consumption of red meat and dairy and subsequently appears on diverse glycans on the cell surface, accumulating mostly on carcinomas. Consequently, humans have circulating anti-Neu5Gc antibodies that play diverse roles in cancer and other chronic inflammation-mediated diseases and that are becoming potential diagnostic and therapeutic targets. Here, we describe a high-throughput sialoglycan microarray assay to assess such anti-Neu5Gc antibodies in the human sera. Neu5Gc-containing glycans and their matched pairs of controls (Neu5Ac-containing glycans), each with a core primary amine, are covalently linked to epoxy-coated glass slides. We exemplify the printing of 56 slides in a 16-well format using a specific nano-printer capable of generating up to 896 arrays per print. Each slide can be used to screen 16 different human sera samples for the evaluation of anti-Neu5Gc antibody specificity, intensity, and diversity. The protocol describes the complexity of this robust tool and provides a basic guideline for those aiming to investigate the response to Neu5Gc dietary carbohydrate antigen in diverse clinical samples in an array format.


Bacillus thuringiensis: a century of research, development and commercial applications.
Bacillus thuringiensis (Bt) is a soil bacterium that forms spores during the stationary phase of its growth cycle. The spores contain crystals, predominantly comprising one or more Cry and/or Cyt proteins (also known as δ-endotoxins) that have potent and specific insecticidal activity. Different strains of Bt produce different types of toxin, each of which affects a narrow taxonomic group of insects. Therefore, Bt toxins have been used as topical pesticides to protect crops, and more recently the proteins have been expressed in transgenic plants to confer inherent pest resistance. Bt transgenic crops have been overwhelmingly successful and beneficial, leading to higher yields and reducing the use of chemical pesticides and fossil fuels. However, their deployment has attracted some criticism particularly with regard to the potential evolution of pest-resistant insect strains. Here, we review recent progress in the development of Bt technology and the countermeasures that have been introduced to prevent the evolution of resistant insect populations.


Strong spin-orbit splitting on bi surfaces.
Using first-principles calculations and angle-resolved photoemission, we show that the spin-orbit interaction leads to a strong splitting of the surface-state bands on low-index surfaces of Bi. The dispersion of the states and the corresponding Fermi surfaces are profoundly modified in the whole surface Brillouin zone. We discuss the implications of these findings with respect to a proposed surface charge density wave on Bi(111) as well as to the surface screening, surface spin-density waves, electron (hole) dynamics in surface states, and to possible applications to the spintronics.


Magnetic resonance imaging in Paget disease of the skull.
Four cases of Paget disease of the skull were studied with magnetic resonance (MR) imaging. With optimal use of projection and technical factors, MR permits simultaneous demonstration of osseous deformity and its relation to the underlying soft tissues. Effects on the brain caused by thickened calvaria and brainstem compression from basilar impression can be detected readily on a single sagittal scan.


Impact of cardiorespiratory rehabilitation program on submaximal exercise capacity of Tunisian male patients with post-COVID19: A pilot study.
Post-COVID19 patients suffer from persistent respiratory, cardiovascular, neurological, and musculoskeletal health complaints such as dyspnea, chest pain/discomfort, and fatigue. In Tunisia, the potential benefits of a cardiorespiratory rehabilitation program (CRRP) after COVID19 remain unclear. The main aim of this study was to evaluate the impact of a CRRP on submaximal exercise capacity, evaluated through the 6-min walk test (6MWT) data in post-COVID19 Tunisian patients. This was a cross-sectional study including 14 moderate to severe COVID19 patients aged from 50 to 70 years. CRRP was performed after the end of patients' hospitalization in COVID19 units for extensive or severe extents of COVID19. Dyspnea (modified medical research council), spirometry data, handgrip strength values, 6MWT data, and 6-min walk work (i.e., 6-min walk distance x weight) were evaluated 1-week pre-CRRP, and 1-week post-CRRP. CRRP included 12 sessions [3 sessions (70 min each)/week for 4 weeks]. Exercise-training included aerobic cycle endurance, strength training, and educational sessions. Comparing pre- and post- CRRP results showed significant improvements in the means±standard deviations of dyspnea by 1.79 ± 0.80 points (p < 0.001), forced expiratory volume in one second by 110 ± 180 ml (p = 0.04), 6-min walk distance by 35 ± 42 m (p = 0.01), 6-min walk work by 2,448 ± 3,925 mkg (p = 0.048), resting heart-rate by 7 ± 9 bpm (p = 0.02) and resting diastolic blood pressure by 6 ± 10 mmHg (p = 0.045). In Tunisia, CRRP seems to improve the submaximal exercise capacity of post-COVID19 patients, mainly the 6-min walk distance and work.


Near infrared fluorescent chlorophyll nanoscale liposomes for sentinel lymph node mapping.
Sentinel lymph node (SLN) mapping using in vivo near infrared fluorescence imaging has attracted great attention during the past few years. Here we report on the early use of poorly water-soluble chlorophyll with near infrared fluorescence extracted from the leaf of Chimonanthus salicifolius, for mouse axillary SLN mapping. To improve the water solubility and SLN targeting of the chlorophyll, we encapsulated the chlorophyll in nanoscale liposomes. The liposome-coated chlorophyll nanocomposites obtained were spherical in shape and had an average diameter of 21.7 ± 6.0 nm. The nanocomposites dispersed well in water, and in aqueous suspension they exhibited brighter near infrared fluorescence than chlorophyll alone. After incubation of the nanocomposites with normal liver cells (QSG-7701) and macrophage cells (Ana-1) for no more than 48 hours, there was no obvious reduction in cell viability. When the nanocomposites were injected intradermally into the paw of a mouse, the axillary SLN was found to be strongly fluorescent and was easily visualized in real time without a requirement for surgery. The intensity of the near infrared fluorescence emitted by the SLN was obviously brighter than that emitted by the SLN of another mouse that had been intradermally injected with chlorophyll alone. Our data show that the liposome-coated chlorophyll nanocomposites could have great potential for clinical SLN mapping due to their lack of toxicity, bright near infrared fluorescence, and small diameter.


Requests for Communication Repair Produced by Typically Developing Preschool-Age Children.
Purpose When breakdowns in communication occur, children may request a repair to increase understanding of the message. Unrepaired communication breakdowns may cause confusions, limit conversational exchanges, and restrict children's learning opportunities. Relatively little is known regarding the conditions under which children produce repair requests. Thus, this study examined the verbal communication repair requests produced by typically developing children and evaluated the relationship between children's repair requests and performance on a theory of mind task. Method Participants included 25 typically developing 4-year-old children who completed standardized, norm-referenced assessments of their expressive language, receptive language, and IQ; a theory of mind task; and a shared book reading task. In the shared book reading task, the examiner elicited child repair requests using questions and statements that contained insufficient information. Results Participants produced a statistically significant greater number of repair requests when presented with questions with insufficient information compared to statements with insufficient information ( p < .001). There were no differences in total repair requests between participants who passed or failed the theory of mind task ( p = .45). Conclusion Among preschool-age children, the shared book reading task provided a naturalistic medium that facilitated the examination of children's repair requests. Results from this study provide baseline information to which the repair requests produced by other populations, such as children with autism spectrum disorder, can be compared.


Classification of Iris Colors and Patterns in Koreans.
The iris is the part of the eye that is colored, and it is an important feature in making an artificial eye. To address the lack of information about the brown Korean irises, we classified the colors and patterns of the irises of 100 Korean eyes. The iris colors were extracted from 100 different digital photographs of each eye using K-mean clustering image analysis. The RGB data of the extracted colors were converted to CIELAB color space data and analyzed to determine the representative color of the iris in each photograph. The color differences were calculated to form groups of similar colors by matching the 100 samples into several groups with minimum color differences among them. The average hue value for each group was calculated, and the corresponding representative color was selected from 2 to 3 colors in one group. The iris colors were classified into seven brown color groups with 2 to 3 colors in each group. The coloration of Koreans' eyes appears to vary in a similar hue with differences in chroma and lightness. In addition, the iris patterns were classified into three basic patterns, namely, crown, sunshine, and flower, as well as three complex patterns that are mixtures of two basic patterns. We classified Korean iris colors and patterns for application in the design of artificial eyes.