Thermostable Branched-Chain Amino Acid Transaminases From the Archaea Geoglobus acetivorans and Archaeoglobus fulgidus: Biochemical and Structural Characterization.
Two new thermophilic branched chain amino acid transaminases have been identified within the genomes of different hyper-thermophilic archaea, Geoglobus acetivorans, and Archaeoglobus fulgidus. These enzymes belong to the class IV of transaminases as defined by their structural fold. The enzymes have been cloned and over-expressed in Escherichia coli and the recombinant enzymes have been characterized both biochemically and structurally. Both enzymes showed high thermostability with optimal temperature for activity at 80 and 85°C, respectively. They retain good activity after exposure to 50% of the organic solvents, ethanol, methanol, DMSO and acetonitrile. The enzymes show a low activity to (R)-methylbenzylamine but no activity to (S)-methylbenzylamine. Both enzymes have been crystallized and their structures solved in the internal aldimine form, to 1.9 Å resolution for the Geoglobus enzyme and 2.0 Å for the Archaeoglobus enzyme. Also the Geoglobus enzyme structure has been determined in complex with the amino acceptor α-ketoglutarate and the Archaeoglobus enzyme in complex with the inhibitor gabaculine. These two complexes have helped to determine the conformation of the enzymes during enzymatic turnover and have increased understanding of their substrate specificity. A comparison has been made with another (R) selective class IV transaminase from the fungus Nectria haematococca which was previously studied in complex with gabaculine. The subtle structural differences between these enzymes has provided insight regarding their different substrate specificities.


Inhibitory effects of lutein on endotoxin-induced uveitis in Lewis rats.
Lutein deposits in the macula and lens of human eyes with high concentration and is well known as an eye-protective nutrient for its beneficial effects on eye disease such as age-related macular degeneration and cataract. The purpose of the present study was to investigate the effects of lutein on endotoxin-induced uveitis (EIU) in rats. EIU was induced in male Lewis rats by subcutaneous injection of 200 microg lipopolysaccharide. Lutein or dexamethasone was administered intravenously at 30 minutes before, at the same time as, and at 30 minutes after LPS treatment. The aqueous humor was collected at 24 hours after LPS injection, the number of infiltrating cells, the protein concentration, and the levels of nitric oxide (NO), tumor necrosis factor (TNF)-alpha, interleukin (IL)-6, prostaglandin (PG)-E2, monocyte chemoattractant protein-1 (MCP-1), and macrophage inflammatory protein (MIP)-2 in the aqueous humor were determined. Immunohistochemical staining with a monoclonal antibody against activated nuclear factor (NF)-kappaB was performed to evaluate the effect of lutein on NF-kappaB activation in the iris-ciliary body (ICB) of rats. A mouse macrophage cell line (RAW264.7 cells) was stimulated with LPS in the presence or absence of lutein. Expression of inducible NO synthase (iNOS), cyclooxygenase-2 (COX-2), and degradation of inhibitor kappaB (IkappaB) were analyzed by Western blot analysis. Lutein suppressed the development of EIU in a dose-dependent fashion. The anti-inflammatory effect of 100 mg/kg lutein was as strong as that of 1 mg/kg dexamethasone. Treatment with lutein reduced the concentrations of NO, TNF-alpha, IL-6, PGE2, MCP-1, and MIP-2 in aqueous humor. Lutein also suppressed the activation of NF-kappaB in the ICB as well as iNOS and COX-2 expression and IkappaB degradation in RAW cells. These findings indicate that lutein has anti-inflammatory effects on EIU by inhibiting the NF-kappaB dependent signaling pathway and the subsequent production of proinflammatory mediators.


Prioritizing Potentially Druggable Mutations with dGene: An Annotation Tool for Cancer Genome Sequencing Data.
A major goal of cancer genome sequencing is to identify mutations or other somatic alterations that can be targeted by selective and specific drugs. dGene is an annotation tool designed to rapidly identify genes belonging to one of ten druggable classes that are frequently targeted in cancer drug development. These classes were comprehensively populated by combining and manually curating data from multiple specialized and general databases. dGene was used by The Cancer Genome Atlas squamous cell lung cancer project, and here we further demonstrate its utility using recently released breast cancer genome sequencing data. dGene is designed to be usable by any cancer researcher without the need for support from a bioinformatics specialist. A full description of dGene and options for its implementation are provided here.


Nuclear magnetic resonance relaxometry as a spectroscopic probe of the coordination sphere of a paramagnetic metal bound to a humic acid mixture.
Protons on water molecules are strongly affected by paramagnetic ions. Since the acid-base properties of water facilitate rapid proton exchange, a single proton nuclear magnetic resonance (NMR) signal is seen in aqueous solutions of paramagnetic ions. Proton relaxation times are significantly affected by paramagnetic species and the readily detectable single signal serves as a powerful amplifier of the information contained concerning the protons in the paramagnetic environment. Where water molecules coordinated to free paramagnetic ions and to metal complexes of ligands that form non-labile (on the NMR time scale) complexes, the effects on water in the two environments can be distinguished. This can provide information on the nature of the ligand binding sites. The example of Cu2+ bound to the Laurentian humic acid mixture reported here using convenient low field NMR relaxometers shows that the information can enrich our understanding of complexation and speciation in the presence of complex mixture ligands characteristic of natural water systems. In this case, the data underline the role of aggregation and conformation in defining the complexation sites.


Effect of topographical distribution of α-synuclein pathology on TDP-43 accumulation in Lewy body disease.
It has been reported that the development of TDP-43 pathology in cases of Lewy body disease (LBD) might be associated with the severity of tau pathology. However, the impact of α-synuclein pathology on TDP-43 accumulation in LBD remains unclear. To clarify whether α-synuclein pathology has an effect on TDP-43 accumulation, independent of tau pathology, we examined by immunohistochemistry 56 cases of LBD using a phosphorylation-dependent TDP-43 antibody. The frequency of TDP-43 pathology in all LBD cases was 18% (10/56). In 37 LBD cases with no or low tau burden (LBD-Ltau; Braak NFT stages 0-II), the frequency of TDP-43 pathology was 19% (7/37). The frequency of TDP-43 pathology in diffuse neocortical type LBD-Ltau cases was 36% (4/11), which was higher than those in limbic and brain stem-predominant types (11-14%). The amygdala and entorhinal cortex were the most frequently affected sites of TDP-43 pathology in LBD-Ltau cases. In LBD-Ltau cases, the proportion of diffuse neocortical type LBD was higher in the TDP-43-positive cases, than that in TDP-43-negative cases (57 vs. 23%). In all LBD cases, α-synuclein pathology in the temporal cortex was significantly more severe in TDP-43-positive cases, and significantly correlated with the severity of TDP-43 pathology in the amygdala. In a multivariate model, the presence of severe α-synuclein pathology was significantly associated with the development of TDP-43 pathology independent of age at death and tau pathology. In the amygdala, TDP-43 was often colocalized with α-synuclein or tau. Given these findings, we suggest that α-synuclein pathology is associated with TDP-43 accumulation in LBD cases.


Prevalence and relative risk of other autoimmune diseases in subjects with autoimmune thyroid disease.
Common autoimmune disorders tend to coexist in the same subjects and to cluster in families. We performed a cross-sectional multicenter study of 3286 Caucasian subjects (2791 with Graves' disease; 495 with Hashimoto's thyroiditis) attending UK hospital thyroid clinics to quantify the prevalence of coexisting autoimmune disorders. All subjects completed a structured questionnaire seeking a personal and parental history of common autoimmune disorders, as well as a history of hyperthyroidism or hypothyroidism among parents. The frequency of another autoimmune disorder was 9.67% in Graves' disease and 14.3% in Hashimoto's thyroiditis index cases (P=.005). Rheumatoid arthritis was the most common coexisting autoimmune disorder (found in 3.15% of Graves' disease and 4.24% of Hashimoto's thyroiditis cases). Relative risks of almost all other autoimmune diseases in Graves' disease or Hashimoto's thyroiditis were significantly increased (>10 for pernicious anemia, systemic lupus erythematosus, Addison's disease, celiac disease, and vitiligo). There was relative "clustering" of Graves' disease in the index case with parental hyperthyroidism and of Hashimoto's thyroiditis in the index case with parental hypothyroidism. Relative risks for most other coexisting autoimmune disorders were markedly increased among parents of index cases. This is one of the largest studies to date to quantify the risk of diagnosis of coexisting autoimmune diseases in more than 3000 index cases with well-characterized Graves' disease or Hashimoto's thyroiditis. These risks highlight the importance of screening for other autoimmune diagnoses if subjects with autoimmune thyroid disease present with new or nonspecific symptoms.


Vascular anatomy in children with pulmonary hypertension regarding the transcatheter Potts shunt.
To morphometrically characterise the region of adjacent descending aorta (DAo) and left pulmonary artery (LPA) regarding the transcatheter creation of the reverse Potts shunt. Retrospective review of the invasive haemodynamic data and measurements of the vessel diameters, distances and angles based on the thoracic CT of children with idiopathic pulmonary arterial hypertension (PAH) with pulmonary-to-systemic systolic pressure ratio ≥0.5. Forty-eight CT scans from 47 patients were analysed. Independent of the PAH severity, the diameters of DAo and LPA, and the area of tightest contact between these vessels were very similar in patients with either infrasystemic or isosystemic/suprasystemic PAH. For total population, the tightest contact area (mean±SD, 51.8±31.9 mm2, range 12.5-177.7 mm2) had an elliptic shape stretched along the DAo length and LPA width. The shortest mean DAo-LPA distance was 1.7±0.8 mm (range 1-5 mm). Only one patient, from the suprasystemic PAH group, had the DAo-LPA distance >4 mm. None had lung tissue identified between these two vessels, while in four patients (8.3%) the prominent bronchial artery was seen coursing exactly between the LPA and DAo. The difference of prevalence of the bronchial arteries between two vessels in patients with either infrasystemic PAH or isosystemic/suprasystemic PAH did not reach statistical significance. Children with idiopathic PAH showed no complicating anatomic or morphometric parameters of the region with adjacent DAo and LPA, which potentially determine the planning of the transcatheter creation of Potts shunt. It holds promises for standardisation of the procedure in the future.


Accelerated thymic maturation and autoreactive T cells in bronchopulmonary dysplasia.
Bronchopulmonary dysplasia (BPD), a chronic lung disease of newborns triggered by oxygen and barotrauma, is characterized by arrested alveolarization. Increased levels of bombesin-like peptides shortly after birth mediate lung injury: anti-bombesin antibody 2A11 protects against BPD in two baboon models. The role of adaptive immunity in BPD has not been explored previously. Our goal was to test the hypothesis that thymic architecture and/or T-cell function is altered with BPD, leading to autoimmunity and immunodeficiency. Thymic structure was analyzed by histopathology of thymic architecture and immunohistochemistry for thymic maturation markers (terminal deoxynucleotidyl transferase, proliferating cell nuclear antigen, CD4, and CD8). Thymic cortical epithelial cells (nurse cells) were studied using HLA-DR and protein gene product 9.5 as markers. Functional analysis was performed with "mixed lymphocyte reaction" of thymocyte or splenocyte responder cells with autologous lung cells as the stimulators. 2A11 treatment attenuates thymic cortical involution in BPD animals, sustaining terminal deoxynucleotidyl transferase-positive prothymocytes and thymocyte proliferation. BPD animals have increased CD4(+) cells in thymic cortex and lung interstitium, which are reduced by 2A11. Conversely, cortical protein gene product 9.5/HLA-DR-positive thymic nurse cells are depleted in BPD animals, but are preserved by 2A11-treatment. Whereas fetal thymocytes and splenocytes respond to phythemagglutinin/ionomycin and to a lesser extent, to autologous lung, BPD thymocytes and splenocytes are phythemagglutinin/ionomycin-unresponsive, and yet react strongly to autologous lung. The 2A11 normalizes these responses. These observations suggest that bombesin-like peptides mediate premature thymic maturation and thymic nurse-cell depletion, leading to autoreactive T cells that could contribute to lung injury.


Optimal photosynthetic use of light by tropical tree crowns achieved by adjustment of individual leaf angles and nitrogen content.
Theory for optimal allocation of foliar nitrogen (ONA) predicts that both nitrogen concentration and photosynthetic capacity will scale linearly with gradients of insolation within plant canopies. ONA is expected to allow plants to efficiently use both light and nitrogen. However, empirical data generally do not exhibit perfect ONA, and light-use optimization per se is little explored. The aim was to examine to what degree partitioning of nitrogen or light is optimized in the crowns of three tropical canopy tree species. Instantaneous photosynthetic photon flux density (PPFD) incident on the adaxial surface of individual leaves was measured along vertical PPFD gradients in tree canopies at a frequency of 0.5 Hz over 9-17 d, and summed to obtain the average daily integral of PPFD for each leaf to characterize its insolation regime. Also measured were leaf N per area (N(area)), leaf mass per area (LMA), the cosine of leaf inclination and the parameters of the photosynthetic light response curve [photosynthetic capacity (A(max)), dark respiration (R(d)), apparent quantum yield (phi) and curvature (theta)]. The instantaneous PPFD measurements and light response curves were used to estimate leaf daily photosynthesis (A(daily)) for each leaf. Leaf N(area) and A(max) changed as a hyperbolic asymptotic function of the PPFD regime, not the linear relationship predicted by ONA. Despite this suboptimal nitrogen partitioning among leaves, A(daily) did increase linearly with PPFD regime through co-ordinated adjustments in both leaf angle and physiology along canopy gradients in insolation, exhibiting a strong convergence among the three species. The results suggest that canopy tree leaves in this tropical forest optimize photosynthetic use of PPFD rather than N per se. Tropical tree canopies then can be considered simple 'big-leaves' in which all constituent 'small leaves' use PPFD with the same photosynthetic efficiency.


Awareness, treatment, control of diabetes mellitus and the risk factors: survey results from northeast China.
The awareness, treatment and control of diabetes mellitus (DM) can effectively reflect on the social status of diabetes conditions. Although several researchers have investigated the awareness, treatment and control rates of diabetes mellitus in China, little is known about their association with risk factors. This study aims to examine the relationship between risk factors and awareness, treatment and control of diabetes mellitus in northeast China. A cross-sectional survey was conducted in 2012. Multistage stratified random cluster sampling design was used to select participants aged 18 to 79 years old. The analysis was based on a representative sample of 1,854 adult subjects. Multivariable logistic regression analysis was used to examine socio-demographic factors associated with the levels of awareness, treatment and control of diabetes mellitus. The awareness, treatment, and control rates of diabetes mellitus were 64.1%, 52.9% and 44.2%, respectively. In the multivariable logistic regression analysis, family history of diabetes was significantly positively associated with awareness (OR, 2.145; 95% CI, 1.600-2.875) and treatment (OR, 2.021; 95% CI, 1.559-2.619) of diabetes mellitus, while negatively associated with control (OR, 0.671; 95% CI, 0.529-0.951). Cigarette smokers and alcohol drinkers were less likely than non-smokers and non-drinkers to be aware of their blood glucose levels (OR, 0.895, 0.614; 95% CI, 0.659-1.216, 0.446-0.844, respectively). Participants who frequently exercise were more likely to be aware of their diabetic conditions than people who never or rarely exercise (OR, 2.003; 95% CI, 1.513-2.651). We found that the awareness and treatment of diabetes mellitus were positively associated with age and were high in participants with a family history of diabetes and those who exercise frequently, but low for cigarette smokers and alcohol drinkers. Participants with a family history of diabetes had their diabetic condition poorly controlled.


Diagnosis of Helicobacter pylori by invasive test: histology.
The accurate detection of Helicobacter pylori (H. pylori), a major cause of gastric cancer, is essential for managing infected patients. Among various diagnostic methods, histology plays a pivotal role in detecting H. pylori and it also provided more information about the degree of inflammation and associated pathology, such as, atrophic gastritis (AG), intestinal metaplasia (IM), and gastric cancer. The diagnosis of H. pylori could be performed in hematoxylin and eosin (H&E) staining, however the specificity can be improved by special stains such as modified Giemsa, Warthin-Starry silver, Genta, and immunohistochemical (IHC) stains. Thus, at least two kinds of stain methods are recommended for diagnosis in practice; H&E staining is routine and Giemsa stain seems to have advantage over other stains because of its simplicity and consistency. IHC stain may be useful in special situations. However, histology has several limitations, including higher cost, longer turnaround time, dependence on the skills of the operator, and interobserver variability in assessment. Furthermore, the density of H. pylori can vary at different sites, possibly leading to sampling error, and the sensitivity of histology may decrease in patients taking proton pump inhibitor (PPI). The updated Sydney system recommend to take five biopsy specimens from different sites; however if this is not possible, the gastric body greater curvature could be a better site to detect current H. pylori infections, especially in the presence of peptic ulcer bleeding, AG and IM, or gastric cancer. In the presence of peptic ulcer bleeding, histology is also the most reliable test. PPIs can affect the result of histology and should be stopped 2 weeks before testing. Postbiopsy bleeding may be increased in patients with anticoagulation therapy, so careful precautions should be taken.


Clustering of protein families into functional subtypes using Relative Complexity Measure with reduced amino acid alphabets.
Phylogenetic analysis can be used to divide a protein family into subfamilies in the absence of experimental information. Most phylogenetic analysis methods utilize multiple alignment of sequences and are based on an evolutionary model. However, multiple alignment is not an automated procedure and requires human intervention to maintain alignment integrity and to produce phylogenies consistent with the functional splits in underlying sequences. To address this problem, we propose to use the alignment-free Relative Complexity Measure (RCM) combined with reduced amino acid alphabets to cluster protein families into functional subtypes purely on sequence criteria. Comparison with an alignment-based approach was also carried out to test the quality of the clustering. We demonstrate the robustness of RCM with reduced alphabets in clustering of protein sequences into families in a simulated dataset and seven well-characterized protein datasets. On protein datasets, crotonases, mandelate racemases, nucleotidyl cyclases and glycoside hydrolase family 2 were clustered into subfamilies with 100% accuracy whereas acyl transferase domains, haloacid dehalogenases, and vicinal oxygen chelates could be assigned to subfamilies with 97.2%, 96.9% and 92.2% accuracies, respectively. The overall combination of methods in this paper is useful for clustering protein families into subtypes based on solely protein sequence information. The method is also flexible and computationally fast because it does not require multiple alignment of sequences.


Sequelae of Kawasaki disease in adolescents and young adults.
Kawasaki disease is an acute vasculitis of unknown etiology that predominantly affects children <5 years of age. Structural damage to the coronary arteries after the acute, self-limited illness is detected by echocardiography in approximately 25% of untreated patients. The long-term effects of the acute coronary arteritis are unknown. To define the spectrum of clinical disease in young adults that can be attributed to Kawasaki disease in childhood, we performed a retrospective survey of cases reported in the English and Japanese published data of adult coronary artery disease attributed to antecedent Kawasaki disease. The mean age at presentation with cardiac sequelae was 24.7 +/- 8.4 years (range 12 to 39) for the 74 patients identified with presumed late sequelae of Kawasaki disease. Symptoms at the time of presentation with cardiac sequelae included chest pain/myocardial infarction (60.8%), arrhythmia (10.8%) and sudden death (16.2%). These symptoms were precipitated by exercise in 82% of patients. One-third of the patients in whom a chest radiograph was taken had ring calcification. Angiographic findings included coronary artery occlusion (66.1%). Extensive development of collateral vessels was reported in 44.1% of patients. Autopsy findings included coronary artery aneurysms (100%) and coronary artery occlusion (72.2%). The acute vasculitis of Kawasaki disease can result in coronary artery damage and rheologic changes predisposing to thrombus formation or progressive atherosclerotic changes that may remain clinically silent for many years. Coronary artery aneurysms and calcification on chest radiography were unusual features in this group of patients. A history of antecedent Kawasaki disease should be sought in all young adults who present with acute myocardial infarction or sudden death.


Tocilizumab for severe COVID-19 pneumonia: Case series of 5 Australian patients.
To describe the first Australian cases of severe acute respiratory syndrome-coronavirus 2 (SARS-CoV2) disease (COVID-19) pneumonia treated with the interleukin-6 receptor antagonist tocilizumab. Retrospective, open-label, real-world, uncontrolled, single-arm case series conducted in 2 tertiary hospitals in NSW, Australia and 1 tertiary hospital in Victoria, Australia. Five adult male patients aged between 46 and 74 years with type 1 respiratory failure due to COVID-19 pneumonia requiring intensive care unit (ICU) admission and biochemical evidence of systemic hyperinflammation (C-reactive protein greater than 100 mg/L; ferritin greater than 700 μg/L) were administered variable-dose tocilizumab. At between 13 and 26 days follow-up, all patients are alive and have been discharged from ICU. Two patients have been discharged home. Two patients avoided endotracheal intubation. Oxygen therapy has been ceased in three patients. Four adverse events potentially associated with tocilizumab therapy occurred in three patients: ventilator-associated pneumonia, bacteremia associated with central venous catheterization, myositis and hepatitis. All patients received broad-spectrum antibiotics, 4 received corticosteroids and 2 received both lopinavir/ritonavir and hydroxychloroquine. The time from first tocilizumab administration to improvement in ventilation, defined as a 25% reduction in fraction of inspired oxygen required to maintain peripheral oxygen saturation greater than 92%, ranged from 7 hours to 4.6 days. Tocilizumab use was associated with favorable clinical outcome in our patients. We recommend tocilizumab be included in randomized controlled trials of treatment for patients with severe COVID-19 pneumonia, and be considered for compassionate use in such patients pending the results of these trials.


[Breastfeeding and antiinfectious agents].
Not infrequently an infection or an antibiotic treatment needed by a lactating mother leads to medical advice against breastfeeding. However, advising against breastfeeding increases morbi-mortality risk in the infant. Besides there are few anti-infectious agents not compatible with breastfeeding, and in these cases, there is usually an alternative to treat the mother's illness. Thus it is important for health professionals to be informed of best sources where to look for the best treatment for the mother compatible with breastfeeding. This article offers the reader a review of basic pharmacodynamics which influence drug use in human lactation, an alphabetically ordered list of available anti-infectious agents coded by numbers indicating risk level, and some web recommendations for the interested reader. We hope to offer the interested reader a useful tool which may prevent some cases of physician led abandonment of lactation.


Histopathologic study of congenital aural atresia in the human embryo.
We studied the histopathologic features of the temporal bones in a human embryo with unilateral aural atresia. The developmental stage of the embryo was at stage 22 in the Carnegie system, and the estimated ovulation age was 8 weeks. There were severe hypoplastic changes in Meckel's and Reichert's arch cartilages without differentiation of the auditory ossicles, hypoplasia of the tubotympanic recess, and resultant abnormal passing of the facial nerve in the affected ear. Abnormal lateral extension of the cartilaginous otic capsule replaced a posterior half of the middle ear region and seemed to form the so-called atresia plate. The external and middle ears of the unaffected side and the bilateral inner ears were morphologically normal. These findings might explain some parts of the complicated mechanism in the development of middle ear anomalies encountered in surgery for congenital aural atresia.


Neutrophil necrosis and annexin 1 degradation associated with airway inflammation in lung transplant recipients with cystic fibrosis.
Neutrophils sequestered in lower respiratory tract secretions in the inflamed lung may undergo apoptosis and/or necrosis and release toxic cellular contents that can injure airways or parenchyma. This study examined the viability of neutrophils retrieved from the proximal airways of lung transplant recipients with bacterial tracheobronchitis. Integrity and stability of intracellular proteins in neutrophils from proximal airways and peripheral blood from lung transplant recipients with bacterial tracheobronchitis were analyzed via Western blot analysis and determination of neutrophil viability by morphologic appearance and flow cytometry. Neutrophils in tracheobronchial secretions from lung transplant recipients with cystic fibrosis who had normal chest radiographic imaging but bronchoscopic evidence of purulent tracheobronchitis post-transplant were necrotic and associated with degradation of intracellular protein annexin 1. The neutrophil influx was compartmentalized to large airways and not detected in peripheral bronchoalveolar airspaces sampled via bronchoalveolar lavage. Peripheral blood neutrophils from healthy subjects cultured in vitro demonstrated that annexin 1 degradation, particularly to a 33 kDa annexin 1 breakdown product (A1-BP), was associated with neutrophil necrosis, but not apoptosis. Although annexin 1 degradation was not specific to neutrophil necrosis, it was a sensitive marker of intracellular protein degradation associated with neutrophil necrosis. Annexin 1 degradation to 33 kDa A1-BP was not observed in peripheral blood neutrophils from healthy subjects, but annexin 1 appeared to be degraded in peripheral blood neutrophils of lung transplant recipients despite a normal morphologic appearance of these cells. Neutrophils were necrotic from the proximal airways of lung transplant recipients with bacterial tracheobronchitis, and this process may begin when neutrophils are still in the systemic circulation prior to sequestration in inflamed airways. Annexin 1 degradation to 33 kDa A1-BP may be useful as a sensitive marker to detect neutrophil necrosis.


'Meaning in life? Make it as bearable, enjoyable and good as possible!': A qualitative study among community-dwelling aged adults who receive home nursing in the Netherlands.
The population of adults ageing in place and using home-care services is growing rapidly worldwide. Meaning in life (MiL) of this group of clients is relevant for healthcare and social workers. MiL is associated with many positive outcomes, but can be challenging for aged persons. Objective of this study was to explore MiL in daily life of community-dwelling aged persons who receive homecare. A hermeneutic phenomenological approach was followed. Three waves of semi-structured interviews took place among 24 clients of a home-care organisation in the Netherlands between November 2015 and July 2018. Photo-elicitation was part of the interview procedure. Interpretative Phenomenological Analysis and dialogues enhanced understanding. Findings show that participants derived meaning from self, others, environment and living. The process of retaining MiL involved maintaining, adapting and discovering. We conclude that community-dwelling aged adults can draw MiL from many sources. Retaining MiL is interwoven in everyday life and requires continuous adaptation to ever-changing life conditions during later life. Although relevant general themes were sketched in this paper, the importance of each, and the connections between them, vary and come to light at the individual level. The themes in this paper and the cases in the appendices provide insights that may help professionals recognise MiL in their work. Besides listening to the stories of aged adults, person-centred interventions should support aged adult's strategy to retain MiL.


3D Analysis of Facial Morphology of a Colombian Population Compared to Adult Caucasians.
The purpose of this study involves describing the facial morphology of a Colombian population with three-dimensional (3D) imaging, and comparing their facial morphology with the Caucasian to create a database for this ethnicity. The study, which included 135 subjects selected from the Valle University in Cali, Colombia, and 535 Caucasian subjects selected from the FaceBase-Data (1U01DE024449-01), was funded by the National Institute of Dental and Craniofacial Research. All images were taken in the natural head position (NHP) and captured using a stereo-photogrammetric camera system (3dMDface) to obtain a 3D image of each patient. The subjects were between 19 to 31 years of age, with a normal body mass index (BMI), and no craniofacial deformities. All images were plotted and analyzed using the 3dMDVultus software to calculate linear and angular measurements. Standard deviation (SD) and means were calculated for each measurement and analyzed using t-test for different samples. The Hispanic population had wider eyes, more protruded upper and lower lips, wider face, and greater mandibular width. Caucasian females had a more acute full profile, larger middle third, and less protrusive lips. Understanding the facial morphology of different populations would help to establish a better diagnosis and treatment planning for each ethnicity. From this study, the following conclusions may be drawn:1. The mean values of Colombian males showed greater measurements than females in the majority of measurements.2. Caucasian females had a more acute full profile, larger middle and lower third, and less protrusive lips.3. This study showed significant ethnic differences in the linear and angular measurements, showing us the importance of considering these findings in the diagnosis and treatment planning when a Hispanic population is involved. Soft tissue values should reflect the norms according to each ethnic population in order to achieve treatment goals.


Short- and long-term effects of habitat fragmentation differ but are predicted by response to the matrix.
Habitat loss and fragmentation are major threats to biodiversity and ecosystem processes. Our current understanding of the impacts of habitat loss and fragmentation is based largely on studies that focus on either short-term or long-term responses. Short-term responses are often used to predict long-term responses and make management decisions. The lack of studies comparing short- and long-term responses to fragmentation means we do not adequately understand when and how well short-term responses can be extrapolated to predict long-term responses, and when or why they cannot. To address this gap, we used data from one of the world's longest-running fragmentation experiments, The Wog Wog Habitat Fragmentation Experiment. Using data for carabid beetles, we found that responses in the long term (more than 22 yr post-fragmentation ≈22 generations) often contrasted markedly with those in the short term (5 yr post-fragmentation). The total abundance of all carabids, species richness and the occurrence of six species declined in the short term in the fragments but increased over the long term. The occurrence of three species declined initially and continued to decline, whilst another species was positively affected initially but decreased in the long term. Species' responses to the matrix that surrounds the fragments strongly predicted both the direction (increase/decline in occurrence) and magnitude of their responses to fragmentation. Additionally, species' responses to the matrix were somewhat predicted by their preferences for different types of native habitat (open vs. shaded). Our study highlights the degree of the matrix's influence in fragmented landscapes, and how this influence can change over time. We urge caution in using short-term responses to forecast long-term responses in cases where the matrix (1) impacts species' responses to fragmentation (by isolating them, creating new habitat or altering fragment habitat) and (2) is likely to change through time.


[Laser photodynamic therapy with hematoporphyrin derivative in lung cancer].
Photodynamic therapy (PDT) using hematoporphyrin derivative (HpD) was performed in 100 cases with lung cancer. The efficacy and limitation of this therapy were evaluated in this paper. Of these 19 cases were early stage lung cancer (Ia) and there were 16 stage I, 11 stage II, 38 stage III and 16 stage IV. The indications of PDT to obtain complete cure should be limited to cases of early stage lung cancer satisfying the following conditions. The focus should be visible endoscopically. Submucosal tumor invasion should be limited to within the bronchial cartilage. The dose of HpD should be more than 2.5 mg/kg body weight but less than 5mg. The light dose should be more than 180 Joules/cm2 in cases of superficial tumor. In advanced lung cancer cases, PDT was effective to treat the local lesion for the improvement of performance status. Combination PDT with surgery made it possible to increase the indications of surgery or reduce the extent of resection area in some cases.


Only half the transcriptomic differences between resistant genetically modified and conventional rice are associated with the transgene.
Besides the intended effects that give a genetically modified (GM) plant the desired trait, unintended differences between GM and non-GM comparable plants may also occur. Profiling technologies allow their identification, and a number of examples demonstrating that unintended effects are limited and diverse have recently been reported. Both from the food safety aspect and for research purposes, it is important to discern unintended changes produced by the transgene and its expression from those that may be attributed to other factors. Here, we show differential expression of around 0.40% transcriptome between conventional rice var. Senia and Senia-afp constitutively expressing the AFP antifungal protein. Analysis of one-fifth of the regulated sequences showed that around 35% of the unintended effects could be attributed to the process used to produce GM plants, based on in vitro tissue culture techniques. A further ∼15% were event specific, and their regulation was attributed to host gene disruption and genome rearrangements at the insertion site, and effects on proximal sequences. Thus, only around half the transcriptional unintended effects could be associated to the transgene itself. A significant number of changes in Senia-afp and Senia are part of the plant response to stress conditions, and around half the sequences for which up-regulation was attributed to the transgene were induced in conventional (but not transgenic) plants after wounding. Unintended effects might, as such, putatively result in widening the self-resistance characteristics because of the transgene in GM plants.


The quality of written information for parents regarding the management of a febrile convulsion: a randomized controlled trial.
To identify whether providing a new information leaflet for parents regarding the management of a febrile convulsion was more effective in comparison with standard leaflets. Although information leaflets are frequently recommended within healthcare, their quality is often poor. Furthermore, the evidence regarding the effectiveness of leaflets is inconsistent. Few studies have evaluated the effects of providing leaflets that have been developed to improve their quality. Within the specialty of paediatrics, parents are often unprepared regarding the management of febrile convulsion at home and the provision of supplementary leaflets is recommended, despite limited evidence regarding the effectiveness of this approach. There is also limited evidence regarding whether improving the quality of these leaflets leads to better outcomes, e.g. increasing parents' behavioural knowledge. Double-blind randomized controlled trial. One hundred and twenty-six parents of children hospitalized due to benign febrile convulsion were recruited (May 2000-February 2002). Seventy-one parents were randomized to receive the standard care (control leaflet) and 55 parents received the intervention (new leaflet). Data were collected from parents on discharge immediately prior to receiving the intervention and 7-14 days following the intervention via telephone interview. Parents who received the new leaflet found this to be more reassuring and easier to understand than parents who received the control leaflet. No differences between groups were identified regarding the primary outcome, i.e. behavioural knowledge and most of the secondary outcomes, e.g. perceived confidence, state anxiety and satisfaction with the leaflet. Although this study provides modest support for the effectiveness of providing high-quality leaflets, further research is required to determine the best methods for optimizing the effectiveness of leaflets provided at hospital discharge. The quality of leaflets may influence some outcomes, e.g. understanding and reassurance with the written information provided; however, additional strategies to inform parents may be necessary.


WebGestalt: an integrated system for exploring gene sets in various biological contexts.
High-throughput technologies have led to the rapid generation of large-scale datasets about genes and gene products. These technologies have also shifted our research focus from 'single genes' to 'gene sets'. We have developed a web-based integrated data mining system, WebGestalt (http://genereg.ornl.gov/webgestalt/), to help biologists in exploring large sets of genes. WebGestalt is composed of four modules: gene set management, information retrieval, organization/visualization, and statistics. The management module uploads, saves, retrieves and deletes gene sets, as well as performs Boolean operations to generate the unions, intersections or differences between different gene sets. The information retrieval module currently retrieves information for up to 20 attributes for all genes in a gene set. The organization/visualization module organizes and visualizes gene sets in various biological contexts, including Gene Ontology, tissue expression pattern, chromosome distribution, metabolic and signaling pathways, protein domain information and publications. The statistics module recommends and performs statistical tests to suggest biological areas that are important to a gene set and warrant further investigation. In order to demonstrate the use of WebGestalt, we have generated 48 gene sets with genes over-represented in various human tissue types. Exploration of all the 48 gene sets using WebGestalt is available for the public at http://genereg.ornl.gov/webgestalt/wg_enrich.php.


Supraspinatus tendon load during abduction is dependent on the size of the critical shoulder angle: A biomechanical analysis.
Shoulders with supraspinatus (SSP) tears are associated with significantly larger critical shoulder angles (CSA) compared to disease-free shoulders. We hypothesized that larger CSAs increase the ratio of joint shear to joint compression forces (defined as "instability ratio"), requiring substantially increased compensatory supraspinatus loads. A shoulder simulator with simulated deltoid, supraspinatus, infraspinatus/teres minor, and subscapularis musculotendinous units was constructed. The model was configured to represent either a normal CSA of 33° or a CSA characteristic of shoulders with rotator cuff tears (38°), and the components of the joint forces were measured. The instability ratio increased for the 38° CSA compared with the control CSA (33°) for a range of motion between 6° to 61° of thoracohumeral abduction with the largest differences in instability observed between 33° and 37° of elevation. In this range, SSP force had to be increased by 13-33% (15-23 N) to stabilize the arm in space. Our results support the concept that a high CSA can induce SSP overload particularly at low degrees of active abduction.


Characteristics and quality of clinical practice guidelines for depression in adults: a scoping review.
Clinical Practice Guidelines (CPGs) should follow an adequate methodology using an evidence-based approach in order to provide reliable recommendations. However, little is known regarding the quality of CPGs for Depression, which precludes its adequate use by stakeholders and mental health professionals. Thus, the aim of this study was to conduct a scoping review to describe the characteristics and quality of CPGs for Depression in adults. We searched CPGs for Depression in adults in eighteen databases. We included those that were published in English or Spanish between January 2014 and May 2018 and were based on systematic reviews of the evidence. Two independent authors extracted the characteristics, type and number of recommendations, and quality (using the Appraisal of Guidelines for Research and Evaluation-II [AGREE-II]) of each included CPG. We included eleven CPGs, of which 9/11 did not include the participation of patients in the development of the CPG, 4/11 CPGs had a score ≥ 70% in the overall evaluation of AGREE-II, and 3/11 CPGs had a score ≥ 70% in its third domain (rigor of development). In addition, only 5/11 CPGs shared their search strategy, while only 4/11 listed the selected studies they used to reach recommendations, and 7/11 CPGs did not clearly state which methodology they used to translate evidence into a recommendation. Most of evaluated CPGs did not take into account the patient's viewpoints, achieved a low score in the rigor of development domain, and did not clearly state the process used to reach the recommendations. Stakeholders, CPCGs developers, and CPGs users should take this into account when choosing CPGs, and interpreting and putting into practice their issued recommendations.


Increased Axon Initial Segment Length Results in Increased Na+ Currents in Spinal Motoneurones at Symptom Onset in the G127X SOD1 Mouse Model of Amyotrophic Lateral Sclerosis.
Amyotrophic lateral sclerosis (ALS) is a neurodegenerative disease preferentially affecting motoneurones. Transgenic mouse models have been used to investigate the role of abnormal motoneurone excitability in this disease. Whilst an increased excitability has repeatedly been demonstrated in vitro in neonatal and embryonic preparations from SOD1 mouse models, the results from the only studies to record in vivo from spinal motoneurones in adult SOD1 models have produced conflicting findings. Deficits in repetitive firing have been reported in G93A SOD1(high copy number) mice but not in presymptomatic G127X SOD1 mice despite shorter motoneurone axon initial segments (AISs) in these mice. These discrepancies may be due to the earlier disease onset and prolonged disease progression in G93A SOD1 mice with recordings potentially performed at a later sub-clinical stage of the disease in this mouse. To test this, and to explore how the evolution of excitability changes with symptom onset we performed in vivo intracellular recording and AIS labelling in G127X SOD1 mice immediately after symptom onset. No reductions in repetitive firing were observed showing that this is not a common feature across all ALS models. Immunohistochemistry for the Na+ channel Nav1.6 showed that motoneurone AISs increase in length in G127X SOD1 mice at symptom onset. Consistent with this, the rate of rise of AIS components of antidromic action potentials were significantly faster confirming that this increase in length represents an increase in AIS Na+ channels occurring at symptom onset in this model.


Perspectives on epilepsy in people with intellectual disabilities: comparison of family carer, staff carer and clinician score profiles on the Glasgow Epilepsy Outcome Scale (GEOS).
Clinical practice with people with intellectual disability relies heavily upon caregiver report. Crucially, the carer's perspective may depend upon his or her relationship to the patient. We investigated similarities and differences within and between family and paid carers in their reports on the Glasgow Epilepsy Outcome Scale (GEOS), an instrument that quantifies concerns about epilepsy in this population [Epilepsia 42 (2001) 1043]. GEOS forms were available on 186 patients (108 males; mean age 39 years) across 384 primary respondents (141 staff, 83 family, 160 clinicians) and independently completed secondary respondents (67 staff, 36 family). Data were analysed to consider levels of concern as rated bv staff carers, family members and clinicians, and also to consider inter-rater agreement on the concerns raised. There were significant differences in the magnitude of concern on each sub-scale [concerns about seizures, treatment, caring and social impact; range of F(2,171)=9.5-64.7; all P<0.0001]. Post hoc testing revealed that family members scored all sub-scales more highly than staff carers or clinicians, and that staff carers scored more highly than clinicians on all but one sub-scale. Inter-rater agreement between family members was considerably higher (range of r=0.69-0.91) than between staff carers (r=0.30-0.47) across the GEOS sub-scales. Association between staff and family ratings was also modest (r< or = 0.39). It is preferable for the same staff member to complete each administration of the GEOS because of inter-staff variability in reporting of concerns. Families provide a consistent, but more extreme, picture and clinicians generally underestimate the concerns of direct caregivers. However, content of concerns varies relatively little across respondents.


On the probability of violations of Fourier's law for heat flow in small systems observed for short times.
We study the statistical mechanics of thermal conduction in a classical many-body system that is in contact with two thermal reservoirs maintained at different temperatures. The ratio of the probabilities, that when observed for a finite time, the time averaged heat flux flows in and against the direction required by Fourier's Law for heat flow, is derived from first principles. This result is obtained using the transient fluctuation theorem. We show that the argument of that theorem, namely, the dissipation function is, close to equilibrium, equal to a microscopic expression for the entropy production. We also prove that if transient time correlation functions of smooth zero mean variables decay to zero at long times, the system will relax to a unique nonequilibrium steady state, and for this state, the thermal conductivity must be positive. Our expressions are tested using nonequilibrium molecular dynamics simulations of heat flow between thermostated walls.


Effects of prey density, temperature and predator diversity on nonconsumptive predator-driven mortality in a freshwater food web.
Nonconsumptive predator-driven mortality (NCM), defined as prey mortality due to predation that does not result in prey consumption, is an underestimated component of predator-prey interactions with possible implications for population dynamics and ecosystem functioning. However, the biotic and abiotic factors influencing this mortality component remain largely unexplored, leaving a gap in our understanding of the impacts of environmental change on ecological communities. We investigated the effects of temperature, prey density, and predator diversity and density on NCM in an aquatic food web module composed of dragonfly larvae (Aeshna cyanea) and marbled crayfish (Procambarus fallax f. virginalis) preying on common carp (Cyprinus carpio) fry. We found that NCM increased with prey density and depended on the functional diversity and density of the predator community. Warming significantly reduced NCM only in the dragonfly larvae but the magnitude depended on dragonfly larvae density. Our results indicate that energy transfer across trophic levels is more efficient due to lower NCM in functionally diverse predator communities, at lower resource densities and at higher temperatures. This suggests that environmental changes such as climate warming and reduced resource availability could increase the efficiency of energy transfer in food webs only if functionally diverse predator communities are conserved.


Serotonergic dorsal raphe nucleus projections to the cholinergic and noncholinergic neurons of the pedunculopontine tegmental region: a light and electron microscopic anterograde tracing and immunohistochemical study.
The serotonergic dorsal raphe nucleus is considered an important modulator of state-dependent neural activity via projections to cholinergic neurons of the pedunculopontine tegmental nucleus (PPT). Light and electron microscopic analysis of anterogradely transported biotinylated dextran, combined with choline acetyltransferase (ChAT) immunohistochemistry, were employed to describe the synaptic organization of mesopontine projections from the dorsal raphe to the PPT. In a separate set of experiments, we utilized immunohistochemistry for the serotonin transporter (SERT), combined with ChAT immunohistochemistry at the light and electron microscopic levels, to determine whether PPT neurons receive serotonergic innervation. The results of these studies indicate that: (1) anterogradely labeled and SERT-immunoreactive axons and presumptive boutons invest the PPT at the light microscopic level; (2) at the ultrastructural level, dorsal raphe terminals in the PPT pars compacta synapse mainly with dendrites and axosomatic contacts were not observed; (3) approximately 12% of dorsal raphe terminals synapse with ChAT-immunoreactive dendrites; and (4) at least 2-4% of the total synaptic input to ChAT-immunoreactive dendrites is of dorsal raphe and/or serotonergic origin. This serotonergic dorsal raphe innervation may modulate cholinergic PPT neurons during alterations in behavioral state. The role of these projections in the initiation of rapid eye movement (REM) sleep and the ponto-geniculo-occipital waves that precede and accompany REM sleep is discussed.


Metformin versus flutamide in the treatment of metabolic consequences of non-obese young women with polycystic ovary syndrome: a randomized prospective study.
In addition to the reproductive consequences, polycystic ovary syndrome (PCOS) is characterized by a metabolic disorder in which hyperinsulinemia and insulin resistance are central features. The effects and possible benefits from insulin-sensitizing drugs are not well known, especially in non-obese women with PCOS. This study was designed to evaluate the effects of metformin and flutamide on metabolic parameters and insulin resistance in non-obese women with PCOS. Thirty non-obese women newly diagnosed with PCOS and 15 age- and weight-matched healthy volunteers as controls were included in the study. Patients were assigned randomly to receive flutamide 250 mg daily or metformin 850 mg three times daily. Glucose, insulin, insulin resistance, androgen levels and glucose and insulin responses to an oral glucose tolerance tests (OGTT) were assessed before and after a 4-week therapy period. A positive correlation was found between body mass index and insulin level in patients with PCOS and controls. Follicle stimulating hormone, luteinizing hormone, free testosterone and dehydroepiandrosterone sulfate levels decreased significantly, but insulin resistance levels were not changed after flutamide therapy. Body weight, free testosterone, insulin and insulin resistance levels decreased significantly after metformin therapy. In conclusion, metformin treatment improved insulin sensitivity and decreased androgen levels, and flutamide decreased androgen levels but failed to improve insulin sensitivity in the non-obese women with PCOS.


Subjective Symptoms in Patients with Eosinophilic Esophagitis Are Related to Esophageal Wall Thickness and Esophageal Body Pressure.
Endoscopic ultrasonography (EUS) and high-resolution manometry (HRM) can be used in the evaluation of eosinophilic esophagitis (EoE) for frequent symptoms such as dysphagia. However, the role of these examinations is not clear. The aim of this study was to objectively evaluate the subjective symptoms of EoE patients with EUS and HRM. Patients who had endoscopic findings indicative of EoE and matched the number of eosinophil infiltrates used as diagnostic criteria were recruited between September 2018 and August 2019. Evaluable subjects underwent EUS and HRM and completed the Gastrointestinal Symptom Rating Scale (GSRS) questionnaire. The esophageal wall thickness (evaluated with EUS) and HRM parameters between patients with and without symptoms were retrospectively compared. Symptomatic patients were re-examined using EUS and HRM 6 months after treatment. A total of 35 patients (29 males, median age of 49 years) were divided into symptomatic (20 patients) and asymptomatic groups (15 patients). The esophageal wall was thicker, and the distal contractile integral (DCI) values were higher in the symptomatic group (P < 0.001). In addition, DCI values were positively correlated with esophageal wall thickness. After treatment, the GSRS scores showed an improving trend for each item. Esophageal wall thickness and DCI values were significantly decreased (Ps < 0.001). Esophageal wall thickening and increased esophageal body pressure may be involved in subjective symptoms. In addition, treatment may reduce esophageal thickness and pressure along with improvement of subjective symptoms.


Methods for studying mutagenesis and checkpoints in Schizosaccharomyces pombe.
Mutations in genome caretaker genes can induce genomic instability, which are potentially early events in tumorigenesis. Cells have evolved biological processes to cope with the genomic insults. One is a multifaceted response, termed checkpoint, which is a network of signaling pathways to coordinate cell cycle transition with DNA repair, activation of transcriptional programs, and induction of tolerance of the genomic perturbations. When genomic perturbations are beyond repair, checkpoint responses can also induce apoptosis or senescence to eliminate those deleterious damaged cells. Fission yeast, Schizosaccharomyces pombe (S. pombe) has served as a valuable model organism for studies of the checkpoint signaling pathways. In this chapter, we describe methods used to analyze mutagenesis and recombinational repair induced by genomic perturbations, and methods used to detect the checkpoint responses to replication stress and DNA damage in fission yeast cells. In the first section, we present methods used to analyze the mutation rate, mutation spectra, and recombinational repair in fission yeast when replication is perturbed by either genotoxic agents or mutations in genomic caretaker gene such as DNA replication genes. In the second section, we describe methods used to examine checkpoint activation in response to chromosome replication stress and DNA damage. In the final section, we comment on how checkpoint activation regulates mutagenic synthesis by a translesion DNA polymerase in generating a mutator phenotype of small sequence alterations in cells, and how a checkpoint kinase appropriately regulates an endonuclease complex to either prevent or allow deletion of genomic sequences and recombinational repair when fission yeast cells experience genomic perturbation in order to avoid deleterious mutations and maintain cell growth.


Why Studying Urgency Is Urgent.
Bowel urgency, the sudden and immediate need to have a bowel movement, is one of the most widely reported and debilitating symptoms of ulcerative colitis. Urgency has a profound impact on patient well-being, often resulting in patient disengagement from education, employment, and social activities. Although its prevalence correlates with disease activity, it is present in states of both disease flare and remission. Postulated pathophysiologic mechanisms are complex, but urgency is likely a consequence of both acute inflammation and structural sequelae of chronic inflammation. Bowel urgency is not widely incorporated into clinical assessment indices or clinical trial endpoints, despite being a pivotal symptom influencing patient health-related quality of life. Addressing urgency can be challenging owing to the associated embarrassment for patients in volunteering this symptom, and its management can be nuanced in the context of a paucity of specific evidence to target it, independently of disease activity. Explicitly inquiring about urgency and integrating it into a multidisciplinary team combining gastroenterologists, psychological support, and continence services is essential to achieving shared satisfaction from treatment. This article outlines the prevalence of urgency and its impact on the quality of life of patients, describes postulated driving mechanisms, and makes recommendations for its inclusion in clinical care and research.


γδ T cells and CD14+ monocytes are predominant cellular sources of cytokines and chemokines associated with severe malaria.
Severe malaria (SM) is associated with high levels of cytokines such as tumor necrosis factor (TNF), interleukin 1 (IL-1), and interleukin 6 (IL-6). The role of chemokines is less clear, as is their cellular source. In a case-control study of children with SM (n = 200), uncomplicated malaria (UM) (n = 153) and healthy community controls (HC) (n = 162) in Papua, New Guinea, we measured cytokine/chemokine production by peripheral blood mononuclear cells (PBMCs) stimulated with live Plasmodium falciparum parasitized red blood cells (pRBC). Cellular sources were determined. Associations between immunological endpoints and clinical/parasitological variables were tested. Compared to HC and UM, children with SM produced significantly higher IL-10, IP-10, MIP-1βm and MCP-2. TNF and MIP-1α were significantly higher in the SM compared to the UM group. IL-10, IL-6, MIP-1α, MIP-1β, and MCP-2 were associated with increased odds of SM. SM syndromes were associated with distinct cytokine/chemokine response profiles compared to UM cases. TNF, MIP-1β, and MIP-1α were produced predominantly by monocytes and γδ T cells, and IL-10 by CD4(+) T cells. Early/innate PBMC responses to pRBC in vitro are informative as to cytokines/chemokines associated with SM. Predominant cellular sources are monocytes and γδ T cells. Monocyte-derived chemokines support a role for monocyte infiltrates in the etiology of SM.


Factors associated with hypertension among adults in Nepal as per the Joint National Committee 7 and 2017 American College of Cardiology/American Heart Association hypertension guidelines: a cross-sectional analysis of the demographic and health survey 2016.
This study investigated the determinants of hypertension in Nepal according to both the Joint National Committee 7 (JNC7) and the American College of Cardiology/American Heart Association (2017 ACC/AHA) guidelines. Cross-sectional study. This study used data collected from the 2016 Nepal Demographic and Health Survey data. 13 393 weighted adults aged ≥18 years enrolled by a stratified cluster sampling strategy were included in our analysis. The primary outcome was hypertension, which was defined according to JNC7 (systolic blood pressure (SBP) ≥140 mm Hg and/or diastolic blood pressure (DBP) ≥90 mm Hg) and 2017 ACC/AHA guidelines (SBP ≥130 mm Hg and/or DBP ≥80 mm Hg). Antihypertensive medication users were also classified as hypertensive. After descriptive analysis, multilevel logistic regression was applied to obtain ORs. About 21% (n=2827) and 44% (n=5918) of the individuals aged ≥18 years were classified as hypertensive according to the JNC7 and 2017 ACC/AHA guidelines, respectively. Following factors were found to be significantly associated with hypertension according to the 2017 ACC/AHA guideline: ≥70 years (adjusted OR (AOR) 5.2; 95% CI 4.3 to 6.2), 50-69 years (AOR 3.9; 95% CI 3.4 to 4.4) and 30-49 years (AOR 2.7; 95% CI 2.4 to 3.0) age groups, male gender (AOR 1.7; 95% CI 1.6 to 1.9), being overweight/obese (AOR 3.0; 95% CI 2.7 to 3.3), residence in provinces 4 (AOR 1.5; 95% CI 1.2 to 2.0) and 5 (AOR 1.5; 95% CI 1.2 to 1.9). No significant association was identified with household wealth status and ecological regions of residence using the 2017 ACC/AHA guideline. Per both guidelines, multiple factors were associated with hypertension. Public health programme aiming to prevent and control hypertension in Nepal should prioritise these factors and focus on individuals with a higher likelihood of hypertension irrespective of educational level, household wealth status and ecological regions of residence.


Nonresonant background suppression in CARS spectra of dispersive media using phase mismatching.
A nonresonant background suppression technique using coherent cancellation through phase mismatching is discussed and applied for a noncollinear beam configuration. A cuvette structure consisting of a glass, a sample, and a glass layer is regarded. Phase mismatching is shown to be a useful method to suppress nonresonant contributions from cuvette glass walls as well as those originating from the sample. A numerical calculation reveals a limit for the background suppression which can be achieved with this technique. Measurements using ethanol as a sample show the possibility to compensate the nonresonant background originating from the cuvette walls and to effectively suppress the nonresonant contribution in the spectrum of the sample by a factor of 10-50, yielding Lorentzian bands biased by a constant background. Direct measurement of depolarization ratios without interfering nonresonant background is demonstrated for ethanol and shows that this technique can readily be combined with the polarization sensitive CARS technique.


Context and explicit threat cue modulation of the startle reflex: preliminary evidence of distinctions between adolescents with principal fear disorders versus distress disorders.
Anxiety and depression are prevalent, impairing disorders. High comorbidity has raised questions about how to define and classify them. Structural models emphasise distinctions between "fear" and "distress" disorders while other initiatives propose they be defined by neurobiological indicators that cut across disorders. This study examined startle reflex (SR) modulation in adolescents with principal fear disorders (specific phobia; social phobia) (n=20), distress disorders (unipolar depressive disorders, dysthymia, generalised anxiety disorder; post-traumatic stress disorder) (n=9), and controls (n=29) during (a) baseline conditions, (b) threat context conditions (presence of contraction pads over the biceps muscle), and (c) an explicit threat cue paradigm involving phases that signalled safety from aversive stimuli (early and late stages of safe phases; early stages of danger phases) and phases that signalled immediate danger of an aversive stimulus (late stages of danger phases). Adolescents with principal fear disorders showed larger SRs than other groups throughout safe phases and early stages of danger phases. SRs did not differ between groups during late danger phases. Adolescents with principal distress disorders showed attenuated SRs during baseline and context conditions compared to other groups. Preliminary findings support initiatives to redefine emotional disorders based on neurobiological functioning.


Hyperbaric oxygen therapy in plastic surgery: a review article.
The most important effects of hyperbaric oxygen (HBO), for the surgeon, are the stimulation of leukocyte microbial killing, the enhancement of fibroblast replication, and increased collagen formation and neovascularization of ischemic tissue. Preoperative hyperbaric oxygen induces neovascularization in tissue with radionecrosis. Refractory osteomyelitis and necrotizing fasciitis appear to respond to adjunctive hyperbaric oxygen. Crush injury and compartment syndrome appear to benefit through preservation of ATP in cell membranes, which limits edema. Hyperbaric oxygen in burn injury permits shorter hospital stays, a reduced number of surgeries, and less fluid replacement. Skin grafts and flaps are reported to take more completely and more rapidly. The same mechanisms may apply in ischemic problem wounds such as infected diabetic extremities. Contraindications and side effects are described. Hyperbaric oxygen will not heal normal wounds more rapidly but may, under certain circumstances, induce problem wounds to heal more like normal ones.


Dabigatran: will it change clinical practice?
Dabigatran (Pradaxa) is a new oral anticoagulant approved in the United States for the primary prevention of stroke and systemic embolization in patients with nonvalvular atrial fibrillation. It offers clinicians an alternative to warfarin (Coumadin), and it has received considerable interest because of its convenience of use, clinical efficacy, and safety profile. However, it is more expensive, and this may limit its widespread use.


Urea synthesis after oral protein ingestion in man.
In an attempt to assess hepatic functional capacity, hourly urea production, and corresponding serum amino acid concentrations after the ingestion of single protein meals (60, 120, and 240 g of protein) were evaluated in 18 normal subjects and in 8 patients with liver disease. In normal subjects, the relationship between urea production and serum amino acid concentration was linear (urea production in milligram urea nitrogen/kilogram lean body mass/hour = 6.3 times mg amino acid nitrogen/100 ml minus 20.5 SE of the estimate 6.9, r = 0.74, P less than 0.001), and variation of protein intake from 50 to 150 g/day for 3 days before testing did not change this relationship. The patients demonstrated impairment of urea synthesis proportional to the clinical severity of their liver disease. The potential clinical applications of these findings need to be determined.


A developmentally regulated antigen associated with neural cell and process migration.
The distribution of an epitope recognized by the monoclonal antibody JONES has been studied immunohistochemically in the developing nervous system of the rat. In the present report, we survey selected regions of the fetal, postnatal, and adult rat nervous system to test the hypothesis that JONES binding is invariably associated with neural cell migration and axon growth in the developing rat. A series of selected developmental stages extending from embryonic day (E) 9 to adult were used in this investigation. The distribution of JONES binding was examined using indirect immunofluorescence, as well as the immunogold procedure. Particular attention was paid to regions where the positions and timing of cell and axon migrations have been well described for the rat. JONES immunoreactivity first appears at E11-12, when it is localized to the lamina terminalis, the telencephalic-diencephalic junction, the midbrain, and the rhombic lip regions of the cytologically undifferentiated neural tube. In all the regions studied, during embryonic and early postnatal life, the labeling is very intense in the ventricular zone and shows a radial array in the adjacent intermediate and marginal zones. The expression of JONES epitope correlates particularly with times of cell migration in the retina, superior colliculus, cerebellum, and telencephalon and in regions undergoing neurite extension, such as the developing optic tract, the white matter of the cerebellum, the dorsal roots, the trigeminal system, and olfactory nerve. JONES binding becomes progressively restricted in the postnatal period. In the adult brain, immunoreactivity is present only in the retina and cerebellum. In the retina, JONES labeling is present in the outer plexiform layer and optic fiber layer. The labeling in the optic fiber layer extends to the optic nerve head and stops abruptly outside the orbit. In the cerebellum, JONES shows a radially oriented pattern throughout the molecular layer and delineates the cell bodies in the Purkinje cell layer. The only non-neural regions that show JONES immunoreactivity are the adrenal medulla and the kidney glomeruli. We conclude that the antigens recognized by the JONES monoclonal antibody are associated with the migration of subsets of cells and axons within the developing rat nervous system and, consequently, may play a role in conveying selectivity to these processes.


High Rates of Prescribing Antimicrobials for Prophylaxis in Children and Neonates: Results From the Antibiotic Resistance and Prescribing in European Children Point Prevalence Survey.
This study was conducted to assess the variation in prescription practices for systemic antimicrobial agents used for prophylaxis among pediatric patients hospitalized in 41 countries worldwide. Using the standardized Antibiotic Resistance and Prescribing in European Children Point Prevalence Survey protocol, a cross-sectional point-prevalence survey was conducted at 226 pediatric hospitals in 41 countries from October 1 to November 30, 2012. Overall, 17693 pediatric patients were surveyed and 36.7% of them received antibiotics (n = 6499). Of 6818 inpatient children, 2242 (32.9%) received at least 1 antimicrobial for prophylactic use. Of 11899 prescriptions for antimicrobials, 3400 (28.6%) were provided for prophylactic use. Prophylaxis for medical diseases was the indication in 73.4% of cases (2495 of 3400), whereas 26.6% of prescriptions were for surgical diseases (905 of 3400). In approximately half the cases (48.7% [1656 of 3400]), a combination of 2 or more antimicrobials was prescribed. The use of broad-spectrum antibiotics (BSAs), which included tetracyclines, macrolides, lincosamides, and sulfonamides/trimethoprim, was high (51.8% [1761 of 3400]). Broad-spectrum antibiotic use for medical prophylaxis was more common in Asia (risk ratio [RR], 1.322; 95% confidence interval [CI], 1.202-1.653) and more restricted in Australia (RR, 0.619; 95% CI, 0.521-0.736). Prescription of BSA for surgical prophylaxis also varied according to United Nations region. Finally, a high percentage of surgical patients (79.7% [721 of 905]) received their prophylaxis for longer than 1 day. A high proportion of hospitalized children received prophylactic BSAs. This represents a clear target for quality improvement. Collectively speaking, it is critical to reduce total prophylactic prescribing, BSA use, and prolonged prescription.


Is this car looking at you? How anthropomorphism predicts fusiform face area activation when seeing cars.
Anthropomorphism encompasses the attribution of human characteristics to non-living objects. In particular the human tendency to see faces in cars has long been noticed, yet its neural correlates are unknown. We set out to investigate whether the fusiform face area (FFA) is associated with seeing human features in car fronts, or whether, the higher-level theory of mind network (ToM), namely temporoparietal junction (TPJ) and medial prefrontal cortex (MPFC) show a link to anthropomorphism. Twenty participants underwent fMRI scanning during a passive car-front viewing task. We extracted brain activity from FFA, TPJ and MPFC. After the fMRI session participants were asked to spontaneously list adjectives that characterize each car front. Five raters judged the degree to which each adjective can be applied as a characteristic of human beings. By means of linear mixed models we found that the implicit tendency to anthropomorphize individual car fronts predicts FFA, but not TPJ or MPFC activity. The results point to an important role of FFA in the phenomenon of ascribing human attributes to non-living objects. Interestingly, brain regions that have been associated with thinking about beliefs and mental states of others (TPJ, MPFC) do not seem to be related to anthropomorphism of car fronts.


What Drives Task Performance During Animal Fluency in People With Alzheimer's Disease?
Animal fluency is a widely used task to assess people with Alzheimer's disease (AD) and other neurological disorders. The mechanisms that drive performance in this task are argued to rely on language and executive functions. However, there is little information regarding what specific aspects of these cognitive processes drive performance on this task. To understand which aspects of language (i.e., semantics, phonological output lexicon, phonological assembly) and executive function (i.e., mental set shifting; information updating and monitoring; inhibition of possible responses) are involved in the performance of animal fluency in people with AD. Animal fluency data from 58 people with probable AD from the DementiaBank Pittsburgh Corpus were analyzed. Number of clusters and switches were measured and nine word properties (e.g., frequency, familiarity) for each of the correct words (i.e., each word counting toward the total score, disregarding non-animals and repetitions) were determined. Random forests were used to understand which variables predicted the total number of correct words, and conditional inference trees were used to search for interactions between the variables. Finally, Wilcoxon tests were implemented to cross-validate the results, by comparing the performance of participants with scores below the norm in animal fluency against participants with scores within the norm based on a large normative sample. Switches and age of acquisition emerged as the most important variables to predict total number of correct words in animal fluency in people with AD. Cross-validating the results, people with AD whose animal fluency scores fell below the norm produced fewer switches and words with lower age of acquisition than people with AD with scores in the normal range. The results indicate that people with AD rely on executive functioning (information updating and monitoring) and language (phonological output lexicon, not necessarily semantics) to produce words on animal fluency.


Listening to food workers: Factors that impact proper health and hygiene practice in food service.
Foodborne disease is a significant problem worldwide. Research exploring sources of outbreaks indicates a pronounced role for food workers' improper health and hygiene practice. To investigate food workers' perceptions of factors that impact proper food safety practice. Interviews with food service workers in Baltimore, MD, USA discussing food safety practices and factors that impact implementation in the workplace. A social ecological model organizes multiple levels of influence on health and hygiene behavior. Issues raised by interviewees include factors across the five levels of the social ecological model, and confirm findings from previous work. Interviews also reveal many factors not highlighted in prior work, including issues with food service policies and procedures, working conditions (e.g., pay and benefits), community resources, and state and federal policies. Food safety interventions should adopt an ecological orientation that accounts for factors at multiple levels, including workers' social and structural context, that impact food safety practice.


Artifacts in computational optical-sectioning microscopy.
We tested the most complete optical model available for computational optical-sectioning microscopy and obtained four main results. First, we observed good agreement between experimental and theoretical point-spread functions (PSF's) under a variety of imaging conditions. Second, using these PSF's, we found that a linear restoration method yielded reconstructed images of a well-defined phantom object (a 10-microns-diameter fluorescent bead) that closely resembled the theoretically determined, best-possible linear reconstruction of the object. Third, this best linear reconstruction suffered from a (to our knowledge) previously undescribed artifactual axial elongation whose principal cause was not increased axial blur but rather the conical shape of the null space intrinsic to nonconfocal three-dimensional (3D) microscopy. Fourth, when 10-microns phantom beads were embedded at different depths in a transparent medium, reconstructed bead images were progressively degraded with depth unless they were reconstructed with use of a PSF determined at the bead's depth. We conclude that (1) the optical model for optical sectioning is reasonably accurate; (2) if PSF shift variance cannot be avoided by adjustment of the optics, then reconstruction methods must be modified to account for this effect; and (3) alternative microscopical or nonlinear algorithmic approaches are required for overcoming artifacts imposed by the missing cone of frequencies that is intrinsic to nonconfocal 3D microscopy.


A new cationic liposome DNA complex enhances the efficiency of arterial gene transfer in vivo.
An important goal of gene therapy for cardiovascular diseases and cancer is the development of effective vectors for catheter-based gene delivery. Although adenoviral vectors have proven effective for this purpose in animal models, the ability to achieve comparable gene transfer with nonviral vectors would provide potentially desirable safety and toxicity features for clinical studies. In this report, we describe the use of a new cationic DNA-liposome complex using an improved expression vector and lipid, N-(3-aminopropyl)-N, N-dimethyl-2,3-bis(dodecyloxy)-1-propaniminium bromide/dioleyl phosphatidylethanolamine (GAP-DL-RIE/DOPE) to optimize catheter-mediated gene transfer in porcine arteries. The efficiency of this vector was compared to DNA alone, DNA with a previously described cationic liposome complex, (+/-)-N-(2-hydroxyethyl)-N, N-dimethyl-2,3-bis(tetradecyloxy)-1-propanaminium bromide (DMRIE/DOPE), and a replication-defective adenoviral vector in a porcine artery gene transfer model. When used in optimal ratios, GAP-DL-RIE/DOPE liposomes provided a 15-fold higher level of gene expression in arteries compared to DNA alone or DMRIE/DOPE. Gene expression was observed in intimal and medial cells. However, when compared to adenoviral vectors (10(10) pfu/ml), gene expression following GAP-DLRIE/DOPE transfection was approximately 20-fold lower. Following intravenous injection of GAP-DLRIE/DOPE in mice, biochemical, hematological, and histopathological abnormalities were not observed. Significant improvements in the efficacy of arterial gene expression can be achieved by optimization of transfection condition with DNA-liposome complexes in vivo that may prove useful for arterial gene delivery in cardiovascular diseases and cancer.


Anilinopyridinate-supported Ru2x+ (x = 5 or 6) paddlewheel complexes with labile axial ligands.
Five new metal-metal bonded Ru2 compounds are presented and discussed: Ru2(ap)4ONO2 (2), [Ru2(ap)4NCMe][BF4] (3), Ru2(ap)4FBF3 (4), Ru2(ap)4OTf (5), and [Ru2(ap)4OTf][Ag(OTf)2] (6) (ap = 2-anilinopyridinate). All compounds have a (4,0) arrangement of the ap ligands about the Ru-Ru bond and contain one sterically blocked axial site and one site containing a labile ligand. These compounds display some of the shortest Ru-Ru distances known for this class of compounds. We demonstrate a reversible interconversion between compounds 3 and 4 as the MeCN and BF4- ligands are readily displaced. Despite the presence of labile axial ligands, compounds 2-5 remain high spin with an S = 3/2 ground state as determined by EPR spectroscopy.


Evaluation of the Wear of the Pivot Bearing in the Gyro C1E3 Pump.
To estimate the lifetime of the pivot bearing system of the sealless centrifugal Gyro C1E3 pump, pivot bearing wear phenomena of the C1E3 were studied. The pivot bearing system consisted of a male and female pivot made of ceramics and ultrahigh molecular weight polyethylene (UHMWPE), respectively. First, many pumping tests were performed with the C1E3 under various pumping conditions, and the effects of impeller position and fluid on wear were analyzed. Through these preliminary tests, it was found that the wear progress of the pivot bearing consisted of initial wear and stationary wear. Most of this initial wear is caused by the plastic deformation of the polyethylene female pivot. It also was observed that bovine blood was almost comparable to water in its effect on the stationary wear rate at the same rotational speed. Based on these results, a long-term pumping test was performed with the C1E3, and initial and stationary wear rates were determined. At the same time, the maximal loosening distance (LDmax) (permissible total wear) of the C1E3 was determined experimentally from hemolytic and hydraulic performance perspectives. By using experimentally determined parameters the lifetime of the pivot bearing system of the C1E3 pump was estimated for various pumping conditions. The lifetime of the pivot bearing system of the C1E3 was typically 10 years for right ventricular assist, 8 years for left ventricular assist, and 5 years for cardiopulmonary bypass.


Structure and Glycan Binding of a New Cyanovirin-N Homolog.
The HIV-1 envelope glycoprotein gp120 is heavily glycosylated and bears numerous high mannose sugars. These sugars can serve as targets for HIV-inactivating compounds, such as antibodies and lectins, which bind to the glycans and interfere with viral entry into the target cell. We determined the 1.6 Å x-ray structure of Cyt-CVNH, a recently identified lectin from the cyanobacterium Cyanothece(7424), and elucidated its glycan specificity by NMR. The Cyt-CVNH structure and glycan recognition profile are similar to those of other CVNH proteins, with each domain specifically binding to Manα(1-2)Manα units on the D1 and D3 arms of high mannose glycans. However, in contrast to CV-N, no cross-linking and precipitation of the cross-linked species in solution was observed upon Man-9 binding, allowing, for the first time, investigation of the interaction of Man-9 with a member of the CVNH family by NMR. HIV assays showed that Cyt-CVNH is able to inhibit HIV-1 with ∼4-fold higher potency than CV-N(P51G), a stabilized version of wild type CV-N. Therefore, Cyt-CVNH may qualify as a valuable lectin for potential microbicidal use.


Mediterranean diet and physical functioning trajectories in Eastern Europe: Findings from the HAPIEE study.
Unhealthy diet may increase the risk of impaired physical functioning in older age. Although poor diet and limited physical functioning both seem to be particularly common in Eastern Europe, no previous study has assessed the relationship between these two factors in this region. The current analysis examined the association between overall diet quality and physical functioning in Eastern European populations. We used data on 25,504 persons (aged 45-69 years at baseline) who participated in the Health Alcohol and Psychosocial factors in Eastern Europe (HAPIEE) study. Dietary assessment at baseline used food frequency questionnaire, and the overall diet quality was evaluated by the Mediterranean diet score (MDS). Physical functioning (PF) was measured by the physical functioning subscale (PF-10) of the 36-item Short-Form Health Survey at baseline and three subsequent occasions over a 10-year period. The cross-sectional and longitudinal relationships between the MDS and PF were examined simultaneously using growth curve models. Men and women with higher adherence to the Mediterranean diet had significantly better PF at baseline; after multivariable adjustment, the regression coefficient per 1-unit increase in the MDS was 0.39 (95% CI: 0.25, 0.52) in men and 0.50 (0.36, 0.64) in women. However, we found no statistically significant link between baseline MDS and the subsequent slope of PF decline in neither gender; the coefficients were -0.02 (-0.04, 0.00) in men and -0.01 (-0.03, 0.02) in women. Our results do not support the hypothesis that the Mediterranean diet has a substantial impact on the trajectories of physical functioning, although the differences existing at baseline may be related to dietary habits in earlier life.


Production of tumour necrosis factor during murine cutaneous leishmaniasis.
We have assessed the role of tumour necrosis factor-alpha (TNF) during cutaneous leishmaniasis and demonstrated that significant levels of TNF were released by spleen cells from infected mice after in vitro restimulation with Leishmania major promastigotes. Spleen cells from both genetically resistant and genetically susceptible mice were equally capable of producing TNF. After challenge with bacterial endotoxin, TNF activity could also be demonstrated in the serum of L. major-infected mice and the titres correlated with the course of cutaneous disease in susceptible and resistant mice. TNF did not exert a direct leishmanicidal effect in vitro. Furthermore, our study indicated that macrophages are the source of L. major-induced TNF activity and that its elicitation is dependent on the presence of T cells. These findings suggest that TNF acts in concert with other cytokines produced during L. major infection and that its role depends on the composition of T cell subsets and cytokines present.


CACNA1A haploinsufficiency causes cognitive impairment, autism and epileptic encephalopathy with mild cerebellar symptoms.
CACNA1A loss-of-function mutations classically present as episodic ataxia type 2 (EA2), with brief episodes of ataxia and nystagmus, or with progressive spinocerebellar ataxia (SCA6). A minority of patients carrying CACNA1A mutations develops epilepsy. Non-motor symptoms associated with these mutations are often overlooked. In this study, we report 16 affected individuals from four unrelated families presenting with a spectrum of cognitive impairment including intellectual deficiency, executive dysfunction, ADHD and/or autism, as well as childhood-onset epileptic encephalopathy with refractory absence epilepsy, febrile seizures, downbeat nystagmus and episodic ataxia. Sequencing revealed one CACNA1A gene deletion, two deleterious CACNA1A point mutations including one known stop-gain and one new frameshift variant and a new splice-site variant. This report illustrates the phenotypic heterogeneity of CACNA1A loss-of-function mutations and stresses the cognitive and epileptic manifestations caused by the loss of CaV2.1 channels function, presumably affecting cerebellar, cortical and limbic networks.


Diagnosis of asymptomatic natalizumab-associated PML: are we between a rock and a hard place?
The diagnosis of natalizumab-associated progressive multifocal leukoencephalopathy (PML) in multiple sclerosis patients in an asymptomatic stage is crucial since it is associated with better clinical outcome measures. Current diagnostic criteria on PML diagnosis in asymptomatic patients require the detection of JC virus and corresponding imaging findings. Magnetic resonance imaging (MRI) is the most sensitive diagnostic method for these purposes. However, the diagnosis of asymptomatic natalizumab-associated PML based on MRI findings can be challenging particularly in case of inconclusive or negative results on JC virus detection in the cerebrospinal fluid. In this report, we present a case series demonstrating different diagnostic scenarios of asymptomatic PML diagnosis based on MRI findings in combination with inconclusive or negative results on JC virus detection in the cerebrospinal fluid. We discuss the challenges of applying current PML diagnostic criteria in asymptomatic natalizumab-associated PML patients and stress the need for specific diagnostic criteria and guidelines regarding managing these diagnostic dilemmas in order to facilitate an early and correct diagnosis of PML presumably leading to a better clinical outcome.


Probing Atomic Distributions in Mono- and Bimetallic Nanoparticles by Supervised Machine Learning.
Properties of mono- and bimetallic metal nanoparticles (NPs) may depend strongly on their compositional, structural (or geometrical) attributes, and their atomic dynamics, all of which can be efficiently described by a partial radial distribution function (PRDF) of metal atoms. For NPs that are several nanometers in size, finite size effects may play a role in determining crystalline order, interatomic distances, and particle shape. Bimetallic NPs may also have different compositional distributions than bulk materials. These factors all render the determination of PRDFs challenging. Here extended X-ray absorption fine structure (EXAFS) spectroscopy, molecular dynamics simulations, and supervised machine learning (artificial neural-network) method are combined to extract PRDFs directly from experimental data. By applying this method to several systems of Pt and PdAu NPs, we demonstrate the finite size effects on the nearest neighbor distributions, bond dynamics, and alloying motifs in mono- and bimetallic particles and establish the generality of this approach.


Membrane nanodomains in T-cell antigen receptor signalling.
The organization of the T-cell's plasma membrane continues to nourish the curiosity of immunologists, cell biologists and biophysicists. The main reason is the biological and biomedical interest to understand the workings of the cell-cell communication network activated by T-cells during an immune response. The molecular armamentarium of the T-cell plasma membrane helps to identify with high sensitivity, specificity and rapidity antigens from invading microbial pathogens and prepare adequate countermeasures to fend them off, while protecting from attacks against our normal tissues. Many T-cell membrane proteins act as receptors to carry out and finely tune these complex tasks. However, the TCR (T-cell receptor) holds a decisive hegemony for its crucial contribution in steering T-cell function and fate. An emerging notion is that TCR proximal signalling occurs at submicrometre-scale membrane domains. In the present chapter, we discuss the current knowledge on the TCR structure and the associated signal transduction machinery and how the notion of membrane nanodomains has decisively contributed to further understand the molecular basis of T-cell activation.


A uniquely human consequence of domain-specific functional adaptation in a sialic acid-binding receptor.
Most mammalian cell surfaces display two major sialic acids (Sias), N-acetylneuraminic acid (Neu5Ac) and N-glycolylneuraminic acid (Neu5Gc). Humans lack Neu5Gc due to a mutation in CMP-Neu5Ac hydroxylase, which occurred after evolutionary divergence from great apes. We describe an apparent consequence of human Neu5Gc loss: domain-specific functional adaptation of Siglec-9, a member of the family of sialic acid-binding receptors of innate immune cells designated the CD33-related Siglecs (CD33rSiglecs). Binding studies on recombinant human Siglec-9 show recognition of both Neu5Ac and Neu5Gc. In striking contrast, chimpanzee and gorilla Siglec-9 strongly prefer binding Neu5Gc. Simultaneous probing of multiple endogenous CD33rSiglecs on circulating blood cells of human, chimp, or gorilla suggests that the binding differences observed for Siglec-9 are representative of multiple CD33rSiglecs. We conclude that Neu5Ac-binding ability of at least some human CD33rSiglecs is a derived state selected for following loss of Neu5Gc in the hominid lineage. These data also indicate that endogenous Sias (rather than surface Sias of bacterial pathogens) are the functional ligands of CD33rSiglecs and suggest that the endogenous Sia landscape is the major factor directing evolution of CD33rSiglec binding specificity. Exon-1-encoded Sia-recognizing domains of human and ape Siglec-9 share only approximately 93-95% amino acid identity. In contrast, the immediately adjacent intron and exon 2 have the approximately 98-100% identity typically observed among these species. Together, our findings suggest ongoing adaptive evolution specific to the Sia-binding domain, possibly of an episodic nature. Such domain-specific divergences should also be considered in upcoming comparisons of human and chimpanzee genomes.


Generic Neutravidin Biosensor for Simultaneous Multiplex Detection of MicroRNAs via Electrochemically Encoded Responsive Nanolabels.
Current electrochemical biosensors for multiple miRNAs require tedious immobilization of various nucleic acid probes. Here, we demonstrate an innovative approach using a generic neutravidin biosensor combined with electrochemically encoded responsive nanolabels for facile and simultaneous multiplexed detection of miRNA-21 and miRNA-141. The selectivity of the biosensor arises from the intrinsic properties of the electrochemically encoded responsive nanolabels, comprising biotinylated molecular beacons (biotin-MB) and metal nanoparticles (metal-NPs). The procedure is a simple one-pot assay, where the targeted miRNA causes the opening of biotin-MB followed by capturing of the biotin-MB-metal-NPs by the neutravidin biosensor and simultaneous detection of the captured metal-NPs by stripping square-wave voltammetry (SSWV). The multiplexed detection of miRNA-21 and miRNA-141 is achieved by differentiation of the electrochemical signature (i.e., the peak current) for the different metal-NP labels. The biosensor delivers simultaneous detection of miRNAs with a linear range of 0.5-1000 pM for miRNA-21 and a limit of detection of 0.3 pM (3σ/sensitivity, n = 3), and a range of 50-1000 pM for miRNA-141, with a limit of detection of 10 pM. Furthermore, we demonstrate multiplexed detection of miRNA-21 and miRNA-141 in a spiked serum sample.


Factors related to obstetric third and fourth degree perineal lacerations in a Jamaican cohort.
The aim of this study was to identify significant and modifiable risk factors associated with obstetric third and fourth degree perineal lacerations and to produce recommendations that may reduce their morbidity and prevalence. This is a retrospective case control study performed between March 2004 and March 2008. All patients diagnosed with third and fourth degree perineal lacerations were identified (cases) along with randomly assigned controls who delivered during the same time period. Nineteen cases and 38 controls were identified giving a total of 57 patients. Each patient's hospital record was collected and the data extracted. When analysed for weight greater than or equal to 3.5 kg versus birthweight of less than 3.5 kg, the difference between cases and controls was found to be statistically significant, with a p-value of 0.012. Of the cases, 21% had an operative delivery (forceps or vacuum) whereas only 2.6% of the controls had an operative delivery. This was found to be statistically significant (p = 0.011). This study has shown that the two main factors related to the obstetric third and fourth degree perineal lacerations were babies weighing more than 3.5 kg and the use of forceps or vacuum to assist with deliveries. These high risk patients should be attended to by the most senior staff that is available.


The default network of the human brain is associated with perceived social isolation.
Humans survive and thrive through social exchange. Yet, social dependency also comes at a cost. Perceived social isolation, or loneliness, affects physical and mental health, cognitive performance, overall life expectancy, and increases vulnerability to Alzheimer's disease-related dementias. Despite severe consequences on behavior and health, the neural basis of loneliness remains elusive. Using the UK Biobank population imaging-genetics cohort (n = ~40,000, aged 40-69 years when recruited, mean age = 54.9), we test for signatures of loneliness in grey matter morphology, intrinsic functional coupling, and fiber tract microstructure. The loneliness-linked neurobiological profiles converge on a collection of brain regions known as the 'default network'. This higher associative network shows more consistent loneliness associations in grey matter volume than other cortical brain networks. Lonely individuals display stronger functional communication in the default network, and greater microstructural integrity of its fornix pathway. The findings fit with the possibility that the up-regulation of these neural circuits supports mentalizing, reminiscence and imagination to fill the social void.