Analysis of two cDNA clones encoding the B lymphocyte antigen CD20 (B1, Bp35), a type III integral membrane protein.
Two cDNA clones encoding the pan-B cell CD20 antigen were isolated from a COS cell expression library. The two clones bear identical coding sequences and differ only in the length of the 3' untranslated region. The predicted CD20 sequence is 297 residues long and contains three hydrophobic domains, one of which is long enough to span the membrane twice. COS cells transfected with either CD20 clone express an immunoreactive protein of 33 kD.


Beyond soft chemistry - bulk and surface modifications of polycrystalline lepidocrocite titanate induced by post-synthesis thermal treatment.
While the soft chemistry of layered alkali metal oxides is adequately understood, the effect of the post-synthesis thermal treatment on their structure, composition, and properties has been underexplored. In this article, we thoroughly investigated the bulk and surface modifications of K0.8M0.4Ti1.6O4 (M = Ni, Cu, Zn) lepidocrocite titanate thermally treated within 200 °C above its synthetic temperature under air. This practice was typically employed in e.g., specimen fabrication for physical property measurements. We observed the expansion of the interlayer distance (b/2) accompanied by a reduction in layer charge density. These findings can be explained by the deintercalation of interlayer K+ ions and the loss of intralayer Ti, M, and O species. Meanwhile, the enrichment of potassium and carbonate on the surfaces was evident. The slight differences in dielectric properties of the pellets thermally treated at different temperatures were attributed to the combination of bulk and surface modifications. At 103 Hz and RT-250 °C, the maximum dielectric constants ε'max of ∼104 with the dielectric loss (tan δ) ∼0.9-1.5 were obtained for K0.8Zn0.4Ti1.6O4.


Novel Bivalent 99mTc-Complex with N-Methyl-Substituted Hydroxamamide as Probe for Imaging of Cerebral Amyloid Angiopathy.
Cerebral amyloid angiopathy (CAA) is characterized by the deposition of amyloid aggregates in the walls of the cerebral vasculature. Recently, the development of molecular imaging probes targeting CAA has been attracting much attention. We previously reported the 99mTc-hydroxamamide (99mTc-Ham) complex with a bivalent benzothiazole scaffold as a binding moiety for amyloid aggregates ([99mTc]BT2) and its utility for CAA-specific imaging. However, the simultaneous generation of two radiolabeled complexes derived from the geometric isomers was observed in the 99mTc-labeling reaction. It was recently reported that the complexation reaction of 99Tc with N-methyl-substituted Ham provided a single 99Tc-Ham complex consisting of two N-methylated Ham ligands with marked stability. In this article, we designed and synthesized a novel N-methylated bivalent 99mTc-Ham complex ([99mTc]MBT2) and evaluated its utility for CAA-specific imaging. N-Methyl substitution of [99mTc]BT2 prevented the generation of its isomer in the 99mTc-labeling reaction. Enhanced in vitro stability of [99mTc]MBT2 as compared with [99mTc]BT2 was observed. [99mTc]MBT2 showed very low brain uptake, which is favorable for CAA-specific imaging. An in vitro inhibition assay using β-amyloid aggregates and in vitro autoradiographic examination of brain sections from a Tg2576 mouse and a CAA patient showed a decline in the binding affinity for amyloid aggregates due to N-methylation of the 99mTc-Ham complex. These results suggest that the scaffold of the 99mTc-Ham complex may play important roles in the in vitro stability and the binding affinity for amyloid aggregates.


Legal document assembly system for introducing law students with legal drafting.
In this paper, we present a method for introducing law students to the writing of legal documents. The method uses a machine-readable representation of the legal knowledge to support document assembly and to help the students to understand how the assembly is performed. The knowledge base consists of enacted legislation, document templates, and assembly instructions. We propose a system called LEDAS (LEgal Document Assembly System) for the interactive assembly of legal documents. It guides users through the assembly process and provides explanations of the interconnection between input data and claims stated in the document. The system acts as a platform for practicing drafting skills and has great potential as an education tool. It allows teachers to configure the system for the assembly of some particular type of legal document and then enables students to draft the documents by investigating which information is relevant for these documents and how the input data shape the final document. The generated legal document is complemented by a graphical representation of legal arguments expressed in the document. The system is based on existing legal standards to facilitate its introduction in the legal domain. Applicability of the system in the education of future lawyers is positively evaluated by the group of law students and their TA. The online version contains supplementary material available at 10.1007/s10506-022-09339-2.


A rare cause of myocardial infarction with non-obstructive coronary arteries-case report of ST-segment elevation myocardial infarction caused by a mediastinal mass.
ST-segment elevation myocardial infarction (STEMI) is attributable to an occluded coronary artery in almost 90% of patients. Accordingly, restoration of coronary perfusion as early as possible, preferably with primary percutaneous coronary intervention, is the recommended treatment by the European Society of Cardiology, to maximise myocardial salvage. However, not all cases of STEMI are because of coronary artery occlusion. ST-segment elevation myocardial infarction that occurs in the absence of obstructive coronary artery disease on angiography has been termed myocardial infarction with non-obstructive coronary arteries (MINOCA). A 44-year-old man was admitted with retrosternal chest pain radiating to the left arm and jaw, and electrocardiography showed extensive anterior ST-segment elevation. Emergency coronary angiography showed all three coronary arteries were patent with Thrombolysis in Myocardial Infarction-3 flow and no evidence of dissection or thrombus. The ST-elevation and pain resolved spontaneously. Troponin-T level rose from <3 ng/L on arrival to 549 ng/L at 12 h. Subsequent cardiac magnetic resonance imaging (MRI) showed a structurally normal heart (without late gadolinium enhancement) but detected an incidental large, lobulated (90 × 31 × 71 mm) mediastinal mass containing multiple cysts in the anterior mediastinum with inflammation and oedema of the parietal pericardium. Tissue biopsy confirmed Hodgkin's lymphoma and the patient was initiated on chemotherapy. Some 3% of ST-segment myocardial infarctions occur in the absence of obstructive coronary disease (MINOCA), is more frequent in younger patients. Cardiac MRI is a useful tool to both identify some of the potential causes of MINOCA and also to confirm the diagnosis of infarction. Some 26% of MINOCA patients have significant biochemical evidence of myocardial injury but have a normal cardiac MRI. This case illustrates a very rare cause of myocardial infarction in a young patient with unobstructed coronary arteries, and highlights the need in such cases for further detailed imaging of the myocardium and thorax to establish the diagnosis and initiate appropriate treatment.


E-cigarette use and willingness to smoke: a sample of adolescent non-smokers.
There is little evidence on the consequences of using electronic cigarettes (e-cigarette) in adolescence. With a multiethnic sample of non-smokers, we assessed the relation between e-cigarette use and social-cognitive factors that predict smoking of combustible cigarettes. School-based cross-sectional survey of 2309 high school students (mean age 14.7 years). Participants reported on e-cigarette use and cigarette use; on smoking-related cognitions (smoking expectancies, prototypes of smokers) and peer smoker affiliations; and on willingness to smoke cigarettes. Regression analyses conducted for non-cigarette smokers tested the association between e-cigarette use and willingness to smoke cigarettes, controlling for demographics, parenting, academic and social competence, and personality variables. Structural equation modelling (SEM) analysis tested whether the relation between e-cigarette use and willingness to smoke was mediated through any of the three smoking-related variables. Non-smokers who had used e-cigarettes (18% of the total sample) showed more willingness to smoke cigarettes compared with those who had never used any tobacco product; the adjusted OR was 2.35 (95% CI 1.73 to 3.19). SEM showed that the relation between e-cigarette use and willingness to smoke was partly mediated through more positive expectancies about smoking, but there was also a direct path from e-cigarette use to willingness. Among adolescent non-smokers, e-cigarette use is associated with willingness to smoke, a predictor of future cigarette smoking. The results suggest that use of e-cigarettes by adolescents is not without attitudinal risk for cigarette smoking. These findings have implications for formulation of policy about access to e-cigarettes by adolescents.


Towards MRI temperature mapping in real time-the proton resonance frequency method with undersampled radial MRI and nonlinear inverse reconstruction.
Optimal control of minimally invasive interventions by hyperthermia requires dynamic temperature mapping at high temporal resolution. Based on the temperature-dependent shift of the proton resonance frequency (PRF), this work developed a method for real-time MRI thermometry which relies on highly undersampled radial FLASH MRI sequences with iterative image reconstruction by regularized nonlinear inversion (NLINV). As a first step, the method was validated with use of a temperature phantom and ex vivo organs (swine kidney) subjected to heating by warm water or a pulsed laser source. The temperature maps obtained by real-time PRF MRI demonstrate good accuracy as independently controlled by fiber-optic temperature sensors. Moreover, the dynamic results demonstrate both excellent sensitivity to single laser pulses (20 ms duration, 6 J energy output) and high temporal resolution, i.e., 200 ms acquisition times per temperature map corresponding to a rate of 5 frames per second. In addition, future extensions to in vivo applications were prepared by addressing the breathing-related motion problem by a pre-recorded library of reference images representative of all respiratory states. The proposed method for real-time MRI thermometry now warrants further developments towards in vivo MRI monitoring of thermal interventions in animals.


Discerning the subfibrillar structure of mineralized collagen fibrils: a model for the ultrastructure of bone.
Biomineralization templated by organic molecules to produce inorganic-organic nanocomposites is a fascinating example of nature using bottom-up strategies at nanoscale to accomplish highly ordered multifunctional materials. One such nanocomposite is bone, composed primarily of hydroxyapatite (HA) nanocrystals that are embedded within collagen fibrils with their c-axes arranged roughly parallel to the long axis of the fibrils. Here we discern the ultra-structure of biomimetic mineralized collagen fibrils (MCFs) as consisting of bundles of subfibrils with approximately 10 nm diameter; each one with an organic-inorganic core-shell structure. Through an amorphous calcium phosphate precursor phase the HA nanocrystals were specifically grown along the longitudinal direction of the collagen microfibrils and encapsulated them within the crystal lattice. They intercalated throughout the collagen fibrils such that the mineral phase surrounded the surface of collagen microfibrils forming an interdigitated network. It appears that this arrangement of collagen microfibrils in collagen fibrils is responsible for the observed ultrastructure. Such a subfibrillar nanostructure in MCFs was identified in both synthetic and natural bone, suggesting this is the basic building block of collagen-based hard tissues. Insights into the ultrastructure of mineralized collagen fibrils have the potential to advance our understanding on the biomineralization principles and the relationship between bone's structure and mechanical properties, including fracture toughness mechanisms. We anticipate that these principles from biological systems can be applied to the rational design of new nanocomposites with improved performance.


Relevance of an inverse problem approach to overcome cut-off wavenumbers disparities in infrared stationary Fourier transform spectrometers.
One of the major limitations to the use of infrared focal plane arrays (IRFPAs) in stationary Fourier transform spectrometers (FTSs) comes from the spatial inhomogeneities of the pixel responses, where the inhomogeneities of the cut-off wavenumbers of the pixels can prevail. The hypothesis commonly assumed for FTSs that all the pixels are equivalent is thus inaccurate and results in a degradation of the estimated spectrum, even far from the cut-off wavenumbers. However, if the individual spectral responses of the pixels are measured beforehand, this a priori information can be used in the inversion process to produce reliable spectra. Thus, spatial inhomogeneities are not an obstacle for the use of infrared stationary FTS. This result is illustrated in this paper by numerical simulations, based on a realistic description of an IRFPA.


Human S100A7 Induces Mature Interleukin1α Expression by RAGE-p38 MAPK-Calpain1 Pathway in Psoriasis.
Psoriatic keratinocytes express exaggerated levels of inflammatory cytokines, and show aberrant hyperproliferation and terminal differentiation in the pathogenesis of psoriasis. The antimicrobial protein hS100A7 (psoriasin) has been found highly expressed in psoriatic skin, but the mechanism and physiological function remain largely unknown. We observed that hS100A7 induces mature interleukin 1α (17kDa) expression in normal human epidermal keratinocytes, which is dependent on RAGE-p38 MAPK and calpain-1 as the inhibitors or knockdown of them completely decreased the expression of mature interleukin1α. Then, we proved mS100a7a15, mature IL-1α and calpain-1 were highly expressed in imquimod-induced psoriasis model and mouse IL-17a-neutralizing antibody treatment attenuated mS100a7a15 expression. At last, PD 151746 (calpain-1 inhibitor) treatment decreased epidermal thickness in imquimod-induced psoriasis model. Taken together, our results suggest that mature IL-1α induced by hS100A7 is via RAGE-p38 MAPK and calpain-1 pathway in keratinocyte and this mechanism may play an important role during psoriasis.


Nigrosome 1 imaging in REM sleep behavior disorder and its association with dopaminergic decline.
Rapid eye movement sleep behavior disorder (RBD) patients have a high risk of developing a Parkinsonian disorder, offering an opportunity for neuroprotective intervention. Predicting near-term conversion, however, remains a challenge. Dopamine transporter imaging, while informative, is expensive and not widely available. Here, we investigate the utility of susceptibility-weighted MRI (SWI) to detect abnormalities of the substantia nigra in RBD, and explore their association with striatal dopaminergic deficits. SWI of the substantia nigra was performed in 46 RBD patients, 27 Parkinson's patients, and 32 control subjects. Dorsal nigral hyperintensity (DNH) was scored by two blinded raters, and separately quantified using a semiautomated process. Forty-two RBD patients were also imaged with 123 I-ioflupane single-photon emission computed tomography (DaT SPECT/CT). Consensus visual DNH classification was possible in 87% of participants. 27.5% of RBD patients had lost DNH, compared with 7.7% of control subjects and 96% of Parkinson's patients. RBD patients lacking DNH had significantly lower putamen dopaminergic SPECT/CT activity compared to RBD patients with DNH present (specific uptake ratios 1.89 vs. 2.33, P = 0.002). The mean quantified DNH signal intensity declined in a stepwise pattern, with RBD patients having lower intensity than controls (0.837 vs. 0.877, P = 0.01) but higher than PD patients (0.837 vs. 0.765, P < 0.001). Over one quarter of RBD patients have abnormal substantia nigra SWI reminiscent of Parkinson's, which is associated with a greater dopaminergic deficit. This modality may help enrich neuroprotective trials with early converters.


Multi-disciplinary and pharmacological interventions to reduce post-operative delirium in elderly patients: A systematic review and meta-analysis.
An estimated 80% of older people undergoing surgery develop postoperative delirium (POD) making them a high-risk group. Research in this area is growing fast but there is no established consensus on strategies for POD prevention or management. A systematic review and meta-analysis were conducted to synthesise data on clinical interventions used to reduce POD among older people undergoing elective and emergency surgery. A range of database searches generated 336 papers. A total of 25 studies met the inclusion criteria and were assessed using the Joanna Briggs Institute Critical Appraisal Checklist. The studies were undertaken across the world. This review identified a range of intervention approaches: comparisons between anaesthetic and sedatives agents, medication-specific interventions and multidisciplinary models of care. Results found more consistencies across multidisciplinary interventions than the pharmacological interventions. In pooled analyses, haloperidol (OR 0.74; 95% CI (confidence interval) 0.44, 1.26) was not statistically significantly associated with reduced POD incidence any more than a placebo. There is a need to implement multidisciplinary interventions, as well as collaboration between clinicians on pre- and postoperative care practices regarding pharmacological interventions to more effectively reduce and manage POD in older people.


Ganglioside GM3 sialidase activity in fibroblasts of normal individuals and of patients with sialidosis and mucolipidosis IV. Subcellular distribution and and some properties.
Sensitive assays for the determination of the ganglioside sialidase activity of fibroblast homogenates were established using ganglioside GM3, 3H-labelled in the sphingosine moiety, as a substrate. Ganglioside GM3 sialidase activity was greatly stimulated by the presence of the non-ionic detergent Triton X-100 and was further enhanced by salts such as NaCl; the optimal pH was 4.5. The subcellular localization of this activity was determined by fractionation using free-flow electrophoresis and found to be exclusively associated with the marker for the plasma membrane, but not with that for lysosomes. This Triton-stimulated ganglioside sialidase activity was selectively inhibited by preincubating intact cells in the presence of millimolar concentrations of Cu2+, suggesting that the activity resides on the external surface of the plasma membrane. In normal fibroblasts homogenates, ganglioside GM3 sialidase was also greatly stimulated by sodium cholate. In contrast to the Triton X-100-activated reaction, however, it was not diminished by prior incubation of intact cells in the presence of Cu2+. Only after cell lysis was Cu2+ inhibitory. the cholate-stimulated ganglioside sialidase activity thus paralleled the behaviour of the lysosomal 4-methylumbelliferyl-alpha-D-N-acetylneuraminic acid (4-MU-NeuAc) sialidase. In fibroblasts from sialidosis patients, the cholate-stimulated ganglioside GM3 sialidase activity, but not that of the Triton-activated enzyme, was profoundly diminished. In fibroblasts from patients with mucolipidosis IV (ML IV), both the Triton X-100- and the cholate-stimulated ganglioside GM3 sialidase activities were in the range of normal controls. The Triton-activated enzyme was associated with the plasma membrane in the same manner as in normal cells. Our findings suggest that, in human fibroblasts, there exist two sialidases that degrade ganglioside GM3: one on the external surface of the plasma membrane, and another that is localized in lysosomes and seems identical with the activity that acts on sialyloligosaccharides and 4-MU-NeuAc. As neither activity was found to be deficient in ML IV fibroblasts, our results argue against the hypothesis of a primary involvement of a ganglioside GM3 sialidase in the pathogenesis of ML IV.


Insulin resistance-related genes and advanced left-sided colorectal adenoma.
Insulin resistance has been linked with colorectal neoplasia through a number of mechanistic and observational studies. Allelic variants of genes encoding components of the insulin pathway, including insulin (INS), insulin receptor (INSR), and insulin receptor substrate-1 and insulin receptor substrate-2 (IRS1 and IRS2) have been associated with hyperinsulinemia and insulin resistance and may, therefore, predict susceptibility to colorectal neoplasia. We investigated whether single nucleotide polymorphisms (SNP) in the INS, INSR, IRS1, and IRS2 genes are associated with risk of advanced left-sided colorectal adenoma, a cancer precursor. We analyzed 20 SNPs in a largely Caucasian study population comprising 766 cases with advanced adenomas of the distal colon and 771 controls, all of whom had undergone flexible sigmoidoscopy as part of the screening arm of the Prostate, Lung, Colorectal and Ovarian Cancer Screening Trial. Overall, we found limited evidence for a role of gene variants of the insulin signaling pathway and prevalence of advanced colorectal adenoma. We observed a statistically significant interaction between INSR genotypes and body mass index (BMI) with colorectal adenoma prevalence (P value for global test = 0.003) and suggestion of an interaction between INSR genotypes and glycemic load (P value for global test = 0.06); however, exploration of the interaction of BMI and glycemic load with the individual SNPs in INSR did not suggest a single SNP that may explain the significance of these global tests of interaction and did not yield any consistent patterns. These findings do not provide strong evidence for associations between polymorphic variation in genes of the insulin signaling pathway and advanced left-sided colorectal adenoma. Evidence for interaction between INSR variants and BMI and glycemic load for risk of advanced left-sided colorectal adenoma requires independent confirmation, and genotyping of INSR across a broader region and at greater density may be necessary to fully elucidate the nature of these interactions.


[Threshold values for cost-effectiveness ratio and public funding of medical technologies].
Rising healthcare costs, together with the rapid emergence of new and expensive medical technologies, have facilitated the use of economic analyses for making coverage decisions. The use of cost-effectiveness studies requires an external criterion (threshold value) for the cost-effectiveness ratio, below which funding would be recommended. Although such a threshold reflects the societal value of a full-quality life-year, currently accepted thresholds have been determined arbitrarily. Studies that screened hundreds of cost-effectiveness analyses have found that the most commonly used threshold is $US 50,000 for an additional QALY (Quality Adjusted Life-Year). This figure reflects the estimated cost per QALY to the US Medicare plan for funding a dialysis treatment for patients with chronic renal failure. While healthcare systems throughout the world, as in Israel, have not explicitly declared using a specific threshold for coverage decisions, some countries use an implicit threshold, above which the decision would usually be negative. In the UK and Australia, for instance, the implicit threshold is $US 50,000 to $US 60,000 per QALY. There are several suggestions to set a differential threshold value between countries, associated with their relative wealth, or between diverse disease and treatment characteristics, e.g. higher thresholds for life-saving treatments. Advantages of setting an explicit threshold include improved transparency and consistency of decisions, improved social equity and enhanced public credibility. Draw-backs might be the creation of an excessively mechanical decision-making process, without consideration of other relevant variables, such as severity of disease, existence of alternatives, or the economic burden to the patient. Adoption of a "flexible threshold" approach, in which the threshold is not the exclusive criterion for decision-making, might resolve these weaknesses. Utilization of the threshold concept is likely to expand in the coming years.


Efficacy and Safety of Therapeutic-Dose Heparin vs Standard Prophylactic or Intermediate-Dose Heparins for Thromboprophylaxis in High-risk Hospitalized Patients With COVID-19: The HEP-COVID Randomized Clinical Trial.
Hospitalized patients with COVID-19 are at risk for venous and arterial thromboembolism and death. Optimal thromboprophylaxis dosing in high-risk patients is unknown. To evaluate the effects of therapeutic-dose low-molecular-weight heparin (LMWH) vs institutional standard prophylactic or intermediate-dose heparins for thromboprophylaxis in high-risk hospitalized patients with COVID-19. The HEP-COVID multicenter randomized clinical trial recruited hospitalized adult patients with COVID-19 with D-dimer levels more than 4 times the upper limit of normal or sepsis-induced coagulopathy score of 4 or greater from May 8, 2020, through May 14, 2021, at 12 academic centers in the US. Patients were randomized to institutional standard prophylactic or intermediate-dose LMWH or unfractionated heparin vs therapeutic-dose enoxaparin, 1 mg/kg subcutaneous, twice daily if creatinine clearance was 30 mL/min/1.73 m2 or greater (0.5 mg/kg twice daily if creatinine clearance was 15-29 mL/min/1.73 m2) throughout hospitalization. Patients were stratified at the time of randomization based on intensive care unit (ICU) or non-ICU status. The primary efficacy outcome was venous thromboembolism (VTE), arterial thromboembolism (ATE), or death from any cause, and the principal safety outcome was major bleeding at 30 ± 2 days. Data were collected and adjudicated locally by blinded investigators via imaging, laboratory, and health record data. Of 257 patients randomized, 253 were included in the analysis (mean [SD] age, 66.7 [14.0] years; men, 136 [53.8%]; women, 117 [46.2%]); 249 patients (98.4%) met inclusion criteria based on D-dimer elevation and 83 patients (32.8%) were stratified as ICU-level care. There were 124 patients (49%) in the standard-dose vs 129 patients (51%) in the therapeutic-dose group. The primary efficacy outcome was met in 52 of 124 patients (41.9%) (28.2% VTE, 3.2% ATE, 25.0% death) with standard-dose heparins vs 37 of 129 patients (28.7%) (11.7% VTE, 3.2% ATE, 19.4% death) with therapeutic-dose LMWH (relative risk [RR], 0.68; 95% CI, 0.49-0.96; P = .03), including a reduction in thromboembolism (29.0% vs 10.9%; RR, 0.37; 95% CI, 0.21-0.66; P < .001). The incidence of major bleeding was 1.6% with standard-dose vs 4.7% with therapeutic-dose heparins (RR, 2.88; 95% CI, 0.59-14.02; P = .17). The primary efficacy outcome was reduced in non-ICU patients (36.1% vs 16.7%; RR, 0.46; 95% CI, 0.27-0.81; P = .004) but not ICU patients (55.3% vs 51.1%; RR, 0.92; 95% CI, 0.62-1.39; P = .71). In this randomized clinical trial, therapeutic-dose LMWH reduced major thromboembolism and death compared with institutional standard heparin thromboprophylaxis among inpatients with COVID-19 with very elevated D-dimer levels. The treatment effect was not seen in ICU patients. ClinicalTrials.gov Identifier: NCT04401293.


Multiple Morphometric Assessment of Microglial Cells in Deafferented Spinal Trigeminal Nucleus.
Microglia (MG) are the first cells to react to the abnormal incoming signals that follow an injury of sensory nerves and play a critical role in the development and maintenance of neuropathic pain, a common sequel of nerve injuries. Here we present population data on cell number, soma size, and length of processes of MG in the caudal division of the spinal trigeminal nucleus (Sp5C) in control mice and at the peak of microgliosis (7 days) following unilateral transection of the infraorbital nerve (IoN). The study is performed combining several bias- and assumption-free imaging and stereological approaches with different immunolabeling procedures, with the objective of tackling some hard problems that often hinder proper execution of MG morphometric studies. Our approach may easily be applied to low-density MG populations, but also works, with limited biases, in territories where MG cell bodies and processes form dense meshworks. In controls, and contralaterally to the deafferented side, MG cell body size and shape and branching pattern matched well the descriptions of "resting" or "surveillant" MG described elsewhere, with only moderate intersubject variability. On the superficial laminae of the deafferented side, however, MG displayed on average larger somata and remarkable diversity in shape. The number of cells and the length of MG processes per mm3 increased 5 and 2.5 times, respectively, indicating a net 50% decrease in the mean length of processes per cell. By using specific immunolabeling and cell sorting of vascular macrophages, we found only a negligible fraction of these cells in Sp5C, with no differences between controls and deafferented animals, suggesting that blood-borne monocytes play at most a very limited role in the microgliosis occurring following sensory nerve deafferentation. In sum, here we present reliable morphometric data on MG in control and deafferented trigeminal nuclei using efficient methods that we propose may equally be applied to any morphometric population analysis of these cells under different physiological or pathological conditions.


First record of Borrelia burgdorferi B31 strain in Dermacentor nitens ticks in the northern region of Parana (Brazil).
The aim of this study was to investigate the presence of DNA of Borrelia burgdorferi sensu lato (s.l.) in ticks that feed on horses used for animal traction in rural Jataizinho, Parana, Brazil. Between February and June 2008, a total of 224 ticks was collected of which 75% were identified as Dermacentor nitens and 25% as Amblyomma cajenense. To amplify B. burgdorferi s.l. DNA, the intergenic space region (ISR) between the 5S (rrf) 23S (rrl) rRNA genes was used as targets for nested-PCR. Two ticks of the D. nitens species were positive for B. burgdorferi s.l. Both species showed a fragment of 184 bp, but the sequencing revealed 99.9% homology with the B. burgdorferi sensu stricto (s.s.) strain B31. These results showed, for the first time, the presence of spirochete DNA infecting ticks that parasitize horses used for animal traction, in the rural municipality mentioned. In conclusion, this study opens up promising prospects for determining the infection rate of B. burgdorferi s.s. genospecies or other species in the equine population, as well as the impact of the infection rate on Lyme disease in the state of Parana.


Dispersal of Golgi matrix proteins during mitotic Golgi disassembly.
During mitosis, the mammalian Golgi disassembles into numerous vesicles and larger membrane structures referred to as clusters or remnants. Following mitosis, the vesicles and clusters reassemble to form an intact Golgi in each daughter cell. One model of Golgi biogenesis states that Golgi matrix proteins remain assembled in mitotic clusters and then serve as a template for Golgi reassembly. To test this idea, we performed a 3D-computational analysis of mitotic cells to determine the extent to which these proteins remain in mitotic clusters. As a control we used brefeldin A-induced Golgi disassembly which causes dispersal of Golgi enzymes, but leaves matrix proteins in remnant structures. Unlike brefeldin A-treated cells, in which matrix proteins were clearly sorted from non-matrix proteins, we observed extensive dispersal of matrix proteins in metaphase cells with no evidence of differential sorting of these proteins from other Golgi proteins. The extensive disassembly of matrix proteins argues against their participation in a stable template and supports a self-assembly mode of Golgi biogenesis.


A loss-of-function and H2B-Venus transcriptional reporter allele for Gata6 in mice.
The GATA-binding factor 6 (Gata6) gene encodes a zinc finger transcription factor that often functions as a key regulator of lineage specification during development. It is the earliest known marker of the primitive endoderm lineage in the mammalian blastocyst. During gastrulation, GATA6 is expressed in early cardiac mesoderm and definitive endoderm progenitors, and is necessary for development of specific mesoderm and endoderm-derived organs including the heart, liver, and pancreas. Furthermore, reactivation or silencing of the Gata6 locus has been associated with certain types of cancer affecting endodermal organs. We have generated a Gata6(H2B-Venus) knock-in reporter mouse allele for the purpose of labeling GATA6-expressing cells with a bright nuclear-localized fluorescent marker that is suitable for live imaging at single-cell resolution. Expression of the Venus reporter was characterized starting from embryonic stem (ES) cells, through mouse embryos and adult animals. The Venus reporter was not expressed in ES cells, but was activated upon endoderm differentiation. Gata6(H2B-Venus/H2B-Venus) homozygous embryos did not express GATA6 protein and failed to specify the primitive endoderm in the blastocyst. However, null blastocysts continued to express high levels of Venus in the absence of GATA6 protein, suggesting that early Gata6 transcription is independent of GATA6 protein expression. At early post-implantation stages of embryonic development, there was a strong correlation of Venus with endogenous GATA6 protein in endoderm and mesoderm progenitors, then later in the heart, midgut, and hindgut. However, there were discrepancies in reporter versus endogenous protein expression in certain cells, such as the body wall and endocardium. During organogenesis, detection of Venus in specific organs recapitulated known sites of endogenous GATA6 expression, such as in the lung bud epithelium, liver, pancreas, gall bladder, stomach epithelium, and vascular endothelium. In adults, Venus was observed in the lungs, pancreas, liver, gall bladder, ovaries, uterus, bladder, skin, adrenal glands, small intestine and corpus region of the stomach. Overall, Venus fluorescent protein under regulatory control of the Gata6 locus was expressed at levels that were easily visualized directly and could endure live and time-lapse imaging techniques. Venus is co-expressed with endogenous GATA6 throughout development to adulthood, and should provide an invaluable tool for examining the status of the Gata6 locus during development, as well as its silencing or reactivation in cancer or other disease states.


Behavioral health insurance parity for federal employees.
To improve insurance coverage of mental health and substance-abuse services, the Federal Employees Health Benefits (FEHB) Program offered mental health and substance-abuse benefits on a par with general medical benefits beginning in January 2001. The plans were encouraged to manage care. We compared seven FEHB plans from 1999 through 2002 with a matched set of health plans that did not have benefits on a par with mental health and substance-abuse benefits (parity of mental health and substance-abuse benefits). Using a difference-in-differences analysis, we compared the claims patterns of matched pairs of FEHB and control plans by examining the rate of use, total spending, and out-of-pocket spending among users of mental health and substance-abuse services. The difference-in-differences analysis indicated that the observed increase in the rate of use of mental health and substance-abuse services after the implementation of the parity policy was due almost entirely to a general trend in increased use that was observed in comparison health plans as well as FEHB plans. The implementation of parity was associated with a statistically significant increase in use in one plan (+0.78 percent, P<0.05) a significant decrease in use in one plan (-0.96 percent, P<0.05), and no significant difference in use in the other five plans (range, -0.38 percent to +0.23 percent; P>0.05 for each comparison). For beneficiaries who used mental health and substance-abuse services, spending attributable to the implementation of parity decreased significantly for three plans (range, -201.99 dollars to -68.97 dollars; P<0.05 for each comparison) and did not change significantly for four plans (range, -42.13 dollars to +27.11 dollars; P>0.05 for each comparison). The implementation of parity was associated with significant reductions in out-of-pocket spending in five of seven plans. When coupled with management of care, implementation of parity in insurance benefits for behavioral health care can improve insurance protection without increasing total costs.


The relationship between living arrangements and higher use of hospital care at middle and older ages: to what extent do observed and unobserved individual characteristics explain this association?
Previous research has shown that certain living arrangements, such as living alone, are associated with worse health at older ages. We assessed the association between living arrangements and hospital care use among middle-aged and older adults, and investigated to what extent observed and unobserved individual characteristics explain this association. Longitudinal Finnish registry data for men and women aged 50-89 years were used for the period 1987-2007. The relationship between living arrangements (based on whether an individual lived with a partner, other adults or alone, and whether they lived with minor/adult children) and heavy hospital care use (i.e., having been in hospital for 8 or more days in a year) was studied. First, we applied logistic regression models and linear probability models controlling for observed time-invariant factors (socioeconomic status measured by education, labour force status, and household income; and marital status), and then individual linear probability models with fixed-effects to further account for unobserved time-invariant individual characteristics in the measurement period. Analyses were done separately for 10 year age-groups. In the logistic regression models, men and women who lived alone had higher crude odds of heavy hospital care use than those living only with their partner. These odds ratios were highest for men and women in the youngest age category (50-59 years, 1.72 and 1.36 respectively) and decreased with age. Adjusting for observed time-invariant socioeconomic status attenuated these odds by 14-40%, but adjusting for marital status did not affect the results. Lower odds were observed among adults aged 50-59 years who lived with their partner and (minor or adult) children. But odds were higher for individuals aged 60-79 years who co-resided with their adult children, regardless of whether they lived with a partner. Adjusting for observed time-invariant factors generally did not change these results. After further adjusting for unobserved time-invariant individual characteristics in the individual fixed-effects models, most of these associations largely attenuated or disappeared, particularly for ages 80-89 years. The association between living arrangements and higher use of hospital care at middle and older ages is largely explained by socioeconomic disadvantage and unobserved time-invariant individual characteristics.


Autoimmune disease: is it a disorder of the microenvironment?
Systemic lupus erythematosus (SLE) is a common systemic autoimmune disease that involves several vital organs including the cardiovascular system, joints, and kidneys. The pathology is characterized by accumulation of autoreactive lymphocytes that attack the patients' own tissues, secretion of autoantibodies and deposition of immune complexes in vital organs. Chronic widespread inflammation is the hallmark of SLE and the target of current therapy. According to recent theories, intonating immune circuits of inflammatory cytokines and immune cells constitute highly specialized targets for SLE therapy, which nonetheless consists for the most part of anti-inflammatory medications and cytotoxic drugs. For advanced autoimmune disorders, cell therapy aiming at introducing "healthy" stem cells has been promising, keeping in mind that in its current state, stem cell therapy is reserved for the most advanced diseases refractory to traditional therapy. Ongoing studies in our laboratories examined the role of the bone marrow microenvironment, in particular, mesenchymal stem cells (MSCs) in the etiopathogenesis of SLE. Specifically, we are testing the hypothesis that, in human SLE mouse model, marrow MSCs are defective structurally and functionally. Preliminary data indicate that structural and functional defects in MSC population from an autoimmune mouse model for human SLE may contribute to this pathology and consequently present a target for cell therapy.


Standardized characterization procedures: a necessary support to regulations.
Sustainable procedures for sludge/biosolids management require the development of strategies to reduce the amount of sludge to be handled and to produce higher quality biosolids. Therefore, realistic and enforceable regulations adapted to local situations must be developed, defined outlet procedures established and good quality products guaranteed. To this end, the definition of standardized characterization procedures and guidelines for good practices becomes a necessary support to regulations. To contribute to the development of the EU directives dealing with sludge, the European Committee for Standardization (CEN) has established Technical Committee 308 (TC 308) whose scope is the standardization of methods for sludge characterization and the production of guidelines of good practice. In this paper, the most recent developments of the CEN/TC 308 work for the chemical, biological and physical characterization of sludge/biosolids are discussed in relation to the main different treatment and outlet options.


Reaction of Lines of the Rapid Cycling Brassica Collection and Arabidopsis thaliana to Four Pathotypes of Plasmodiophora brassicae.
The clubroot reaction of five Rapid Cycling Brassica Collection (RCBC) lines (Brassica carinata, B. juncea, B. napus, B. oleracea, and B. rapa) and 84 lines of Arabidopsis thaliana to pathotypes 2, 3, 5, and 6 of Plasmodiophora brassicae (as classified on William's system) was assessed. Also, the reaction of the Arabidopsis lines to a single-spore isolate of each of pathotypes 3 and 6 was compared with that of a field isolate. Seedlings were inoculated with resting spores of P. brassicae, maintained at 25 and 20°C (day and night, respectively), and assessed for clubroot incidence and severity at 6 weeks after inoculation. Several lines of A. thaliana and RCBC exhibited a differential response to pathotype but none of the lines were immune. Among the RCBC lines, B. napus was resistant to all of the pathotypes; B. oleracea was resistant to pathotypes 2, 3, and 5; B. carinata and B. rapa were resistant to pathotypes 2 and 5; and B. juncea was susceptible to pathotypes 5 and 6 and had an intermediate response to pathotypes 2 and 3. Line Ct-1 of A. thaliana was highly resistant to pathotype 2, Pu2-23 was highly resistant to pathotype 5, and Ws-2 and Sorbo were highly resistant to pathotype 6. These results indicate that the lines of RCBC and A. thaliana have potential for use as model crops for a wide range of studies on clubroot, and could be used to differentiate these four pathotypes of P. brassicae. The reaction of the RCBC lines to pathotype 6 was highly correlated with response under field conditions but the reaction to the single-spore isolates of pathotypes 3 and 6 was not strongly correlated with reaction to the field collections in the Arabidopsis lines.


Genetic abnormalities of the EGFR pathway in African American Patients with non-small-cell lung cancer.
Previous studies in non-small-cell lung cancer (NSCLC) have demonstrated a wide variation in responsiveness to epidermal growth factor receptor (EGFR) -targeting agents and in genetic aberrancies of the EGFR pathway according to ethnic background, most notably a higher frequency of activating EGFR mutations among East-Asian patients. We investigated the frequency of EGFR pathway aberrancies among African American patients with NSCLC, for whom limited information presently exists. EGFR fluorescent in situ hybridization (FISH) was performed on archived tissues from 53 African American patients. Extracted DNA was sequenced for mutational analysis of EGFR exons 18 to 21 and KRAS exon 2. Results were compared by multivariate analysis to an historical control cohort of 102 white patients with NSCLC. African Americans were significantly less likely to harbor activating mutations of EGFR than white patients (2% v 17%; P = .022). Only one EGFR mutation was identified, a novel S768N substitution. EGFR FISH assay was more frequently positive for African Americans than for white patients (51% v 32%; P = .018). KRAS mutational frequency did not differ between the groups (23% v 21%; P = .409). African American patients with NSCLC are significantly less likely than white counterparts to harbor activating mutations of EGFR, which suggests that EGFR tyrosine kinase inhibitors (TKIs) are unlikely to yield major remissions in this population. Our findings add to a growing body of evidence that points to genetic heterogeneity of the EGFR pathway in NSCLC among different ethnic groups and that underscores the need for consideration of these differences in the design of future trials of agents that target the EGFR pathway.


Magnesium supplementation for the management of essential hypertension in adults.
Epidemiological evidence on the effects of magnesium on blood pressure is inconsistent. Metabolic and experimental studies suggest that magnesium may have a role in the regulation of blood pressure. To evaluate the effects of magnesium supplementation as treatment for primary hypertension in adults. We searched the Cochrane Library, MEDLINE, EMBASE, Science Citation Index, ISI Proceedings, ClinicalTrials.gov, Current Controlled Trials, CAB abstracts, and reference lists of systematic reviews, meta-analyses and randomised controlled trials (RCTs) included in the review. Inclusion criteria were: 1) RCTs of a parallel or crossover design comparing oral magnesium supplementation with placebo, no treatment, or usual care; 2) treatment and follow-up >/=8 weeks; 3) participants over 18 years old, with raised systolic blood pressure (SBP) >/=140 mmHg or diastolic blood pressure (DBP) >/=85 mmHg; 4) SBP and DBP reported at end of follow-up. We excluded trials where: participants were pregnant; received antihypertensive medication which changed during the study; or magnesium supplementation was combined with other interventions. Two reviewers independently abstracted data and assessed trial quality. Disagreements were resolved by discussion or a third reviewer. Random effects meta-analyses and sensitivity analyses were conducted. Twelve RCTs (n=545) with eight to 26 weeks follow-up met our inclusion criteria. The results of the individual trials were heterogeneous. Combining all trials, participants receiving magnesium supplements as compared to control did not significantly reduce SBP (mean difference: -1.3 mmHg, 95% CI: -4.0 to 1.5, I(2)=67%), but did statistically significantly reduce DBP (mean difference: -2.2 mmHg, 95% CI: -3.4 to -0.9, I(2)=47%). Sensitivity analyses excluding poor quality trials yielded similar results. Sub-group analyses and meta-regression indicated that heterogeneity between trials could not be explained by dose of magnesium, baseline blood pressure or the proportion of males among the participants. In view of the poor quality of included trials and the heterogeneity between trials, the evidence in favour of a causal association between magnesium supplementation and blood pressure reduction is weak and is probably due to bias. This is because poor quality studies generally tend to over-estimate the effects of treatment. Larger, longer duration and better quality double-blind placebo controlled trials are needed to assess the effect of magnesium supplementation on blood pressure and cardiovascular outcomes.


Towards a holistic understanding of the beneficial interactions across the Populus microbiome.
Interactions between trees and microorganisms are tremendously complex and the multispecies networks resulting from these associations have consequences for plant growth and productivity. However, a more holistic view is needed to better understand trees as ecosystems and superorganisms, where many interacting species contribute to the overall stability of the system. While much progress has been made on microbial communities associated with individual tree niches and the molecular interactions between model symbiotic partners, there is still a lack of knowledge of the multi-component interactions necessary for holistic ecosystem-level understanding. We review recent studies in Populus to emphasize the importance of such holistic efforts across the leaf, stem and rooting zones, and discuss prospects for future research in these important ecosystems.


Diagnosis and management of ductal carcinoma in situ (DCIS).
Systematic synthesis of the published evidence about incidence, risk factors, and management options for women with ductal carcinoma in situ (DCIS) of the breast. Original epidemiologic studies were sought from several databases to identity articles published in English between 1970 and January 31, 2009. Incidence of DCIS in the general population and among women at greater risk of breast cancer and patient outcomes after diagnostic magnetic resonance imaging (MRI) or sentinel lymph node biopsy (SLNB) were abstracted into the developed standardized form. Patient outcomes after breast conserving surgery with or without adjuvant radio- or chemotherapy or after mastectomy were compared from randomized controlled clinical trials (RCTs) and observational studies. Three hundred seventy-four publications were eligible for the review. Rarely diagnosed before 1980, the incidence of DCIS increased by 270 percent since 1987 to 37.5 per 100,000 women in 2001, partially due to increased use of mammography with no good evidence of overdiagnosis (63 publications). Incidence was higher with increasing age, breast density, and family history and lower among physically active women and aspirin users (29 publications). Tamoxifen did not prevent DCIS at longer followup in women at high risk of breast cancer (two RCTs). No good evidence was identified around the optimal use of MRI for treatment planning (64 publications). Case-series from academic centers reported that around 5 percent of women with final histological diagnosis of DCIS had positive sentinel nodes and 1 percent were upgraded to metastatic cancer with no significant differences in outcomes (50 publications). Good evidence from five RCTs (ten publications) suggested that breast conserving surgery with adjuvant radiation reduced ipsilateral (the same breast) tumors by 53 percent with no differences in mortality or contralateral (the second breast) cancer. One RCT demonstrated that adjuvant chemotherapy reduced ipsilateral and contralateral cancer. Ten-year post diagnostic survival was more than 98 percent, while the rates of ipsilateral cancer were around 10 percent (133 publications of 64 observational studies). Major risk factors for ipsilateral cancer were younger age, larger tumor size, comedo necrosis, and positive surgical margins. Limited evidence of worse incidence and advanced outcomes in racial subgroups varied across the studies. Inconsistent evidence suggested that Her2 receptor and negative estrogen receptor status were associated with worse outcomes. No good evidence was found that adjuvant chemotherapy or mastectomy can improve outcomes and there was no evidence on natural history of DCIS or on quality of life among women treated for DCIS. Incidence of DCIS continued to increase with no evidence of overdiagnosis or effective preventive strategies. There is a need to better identify problematic lesions from mammography that are most likely to contain some invasive breast cancer. Most prognostic factors for invasive breast cancer are also prognostic factors for DCIS. The role of MRI and SLNB should be investigated as tools to improve pre-surgical decisonmaking and staging. Breast conserving surgery with adjuvant radiotherapy can benefit all women, though the absolute impact may be small for some women. Ongoing trials will shed light on the optimal clinical strategy for treating DCIS.


Multi-point interrogation of FBG sensors using cascaded flexible wavelength-division Sagnac loop filters.
A simple optical signal demodulation technique is demonstrated for multi-point interrogation of fiber Bragg grating sensors. The wavelength-division responses of polarization-maintaining fiber Sagnac loop filters are flexibly controlled to interrogate the simultaneous optical intensity changes from the independent wavelength shifts of multiple fiber Bragg grating sensors. Experimental performance is also demonstrated to show the simultaneous detection of the separated FBG sensing signals by individual photo detectors.


Sex differences in management and outcomes of patients with atrial fibrillation in the Middle East: Gulf survey of atrial fibrillation events (Gulf SAFE).
Differences in the management of atrial fibrillation (AF) between men and women were investigated by using Gulf SAFE data in the Middle East. The study included 2,043 patients presenting with AF to emergency room (ER) were prospectively enrolled and followed for one-year. Women were older, have higher body mass index (BMI), comorbidities, and health complications than men. With regard to management of AF, cardioversion was recommended more often for men (16.7% vs. 9.3%), and underwent electrical cardioversion (2.2% vs. 1.1%). Women were prescribed digoxin more frequently than men (25.6% vs. 17.4%) and a significant number women received warfarin alone (31.1% vs. 8.7%). No difference between the sexes was noticed in One-year rates of stroke/transient ischemic attacks (TIA) and all-cause of mortality after one-year follow-up (3.1% men vs. 3.3% women, and 7.5% vs. 7.4%). Older age (≥ 65 years), smoking, alcohol use, CHADS2 scores ≥5 were some of the significant risk factors in men with AF. Suboptimal use of anticoagulants, higher mortality and stroke/TIA events at one year are high but similar between the sexes. ER management revealed high use of rate control strategy and high rate of hospital admission was noticed in women.


Localised deposition of amyloid in tears of the rotator cuff.
Age-related localised deposition of amyloid in connective tissue has been found in degenerative articular and periarticular tissue. Biopsies of the supraspinatus tendon of 28 patients undergoing repair of the rotator cuff were analysed histologically for the presence of localised deposition of amyloid. There was a long history of impingement in 20 patients, and eight patients had suffered an acute traumatic tear with no preceding symptoms. Localised deposition of amyloid identified by Congo Red staining was detected in 16 samples (57%). Amyloid was present in 14 (70%) of the degenerative tears, but in only two (25%) of the acute tears. Immunohistochemical staining showed that the amyloid deposits were positive for P component, but negative for kappa and lambda light chains, prealbumin, and beta2 microglobulin. Critical electrolyte staining revealed highly-sulphated glycosaminoglycans at sites of deposition of amyloid. The presence of localised deposition of amyloid in tears of the rotator cuff is likely to represent irreversible structural changes. These findings support the theory that impingement and tears are due to intrinsic degenerative changes within the tendons of the rotator cuff.


Complex brain network properties in late L2 learners and native speakers.
Whether the neural mechanisms that underlie the processing of a second language in highly proficient late bilinguals (L2 late learners) are similar or not to those that underlie the processing of the first language (L1) is still an issue under debate. In this study, a group of late learners of Spanish whose native language is English and a group of Spanish monolinguals were compared while they read sentences, some of which contained syntactic violations. A brain complex network analysis approach was used to assess the time-varying topological properties of the functional networks extracted from the electroencephalography (EEG) recording. Late L2 learners showed a lower degree of parallel information transfer and a slower propagation between regions of the brain functional networks while processing sentences containing a gender mismatch condition as compared with a standard sentence configuration. In contrast, no such differences between these conditions were detected in the Spanish monolinguals. This indicates that when a morphosyntactic language incongruence that does not exist in the native language is presented in the second language, the neural activation pattern is configured differently in highly proficient late bilinguals than in monolinguals.


Specific effects of BCL10 Serine mutations on phosphorylations in canonical and noncanonical pathways of NF-κB activation following carrageenan.
To determine the impact of B cell leukemia/lymphoma (BCL) 10 on the phosphorylation of crucial mediators in NF-κB-mediated inflammatory pathways, human colonic epithelial cells were exposed to carrageenan (CGN), a sulfated polysaccharide commonly used as a food additive and known to induce NF-κB nuclear translocation by both canonical and noncanonical pathways. Phosphorylations of intermediates in inflammatory cascades, including NF-κB-inducing kinase (NIK) at Thr(559), transforming growth factor-β-activating kinase (TAK) 1 at Thr(184), Thr(187), and Ser(192), and inhibitory factor κBα (IκBα) at Ser(32), were examined following mutation of BCL10 at Ser(138) and at Ser(218). Specific phosphoantibodies were used for detection by enzyme-linked immunosorbent assay, immunoblot, and confocal microscopy of differences in phosphorylation following transfection by mutated BCL10. Both mutations demonstrated dominant-negative effects, with inhibition of phospho(Ser(32))-IκBα to less than control levels. Both of the BCL10 mutations reduced the CGN-induced increases in nuclear RelA and p50, but only the Ser(138) mutation inhibited the CGN-induced increases in nuclear RelB and p52 and in NIK Thr(559) phosphorylation. Hence, the phosphorylation of BCL10 Ser(138), but not Ser(218), emerged as a critical event in activation of the noncanonical pathway of NF-κB activation. Either BCL10 Ser(138) or Ser(218) mutation inhibited the phosphorylation of TAK1 at Thr(184) and at Thr(187), but not at Ser(192). These findings indicate that BCL10 phosphorylations act upstream of phosphorylations of NIK, TAK1, and IκBα and differentially affect the canonical and noncanonical pathways of NF-κB activation.


MOOCs 101: an introduction to massive open online courses.
Massive Open Online Courses (MOOCs) are a new type of online class that allow anyone, anywhere, to participate via video lectures, computer graded tests, and discussion forums. This article will give a basic overview of what MOOCs are, how they work, and some of their inherent advantages and disadvantages. It will also explore what MOOCs mean for medical education and libraries. A list of MOOC-related resources is also included.


Columnar shifts as symmetry-breaking degrees of freedom in molecular perovskites.
We introduce columnar shifts-collective rigid-body translations-as a structural degree of freedom relevant to the phase behaviour of molecular perovskites ABX3 (X = molecular anion). Like the well-known octahedral tilts of conventional perovskites, shifts also preserve the octahedral coordination geometry of the B-site cation in molecular perovskites, and so are predisposed to influencing the low-energy dynamics and displacive phase transitions of these topical systems. We present a qualitative overview of the interplay between shift activation and crystal symmetry breaking, and introduce a generalised terminology to allow characterisation of simple shift distortions, drawing analogy to the "Glazer notation" for octahedral tilts. We apply our approach to the interpretation of a representative selection of azide and formate perovskite structures, and discuss the implications for functional exploitation of shift degrees of freedom in negative thermal expansion materials and hybrid ferroelectrics.


Wildfire combustion and carbon stocks in the southern Canadian boreal forest: Implications for a warming world.
Boreal wildfires are increasing in intensity, extent, and frequency, potentially intensifying carbon emissions and transitioning the region from a globally significant carbon sink to a source. The productive southern boreal forests of central Canada already experience relatively high frequencies of fire, and as such may serve as an analog of future carbon dynamics for more northern forests. Fire-carbon dynamics in southern boreal systems are relatively understudied, with limited investigation into the drivers of pre-fire carbon stocks or subsequent combustion. As part of NASA's Arctic-Boreal Vulnerability Experiment, we sampled 79 stands (47 burned, 32 unburned) throughout central Saskatchewan to characterize above- and belowground carbon stocks and combustion rates in relation to historical land use, vegetation characteristics, and geophysical attributes. We found southern boreal forests emitted an average of 3.3 ± 1.1 kg C/m2 from field sites. The emissions from southern boreal stands varied as a function of stand age, fire weather conditions, ecozone, and soil moisture class. Sites affected by historical timber harvesting had greater combustion rates due to faster carbon stock recovery rates than sites recovering from wildfire events, indicating that different boreal forest land use practices can generate divergent carbon legacy effects. We estimate the 2015 fire season in Saskatchewan emitted a total of 36.3 ± 15.0 Tg C, emphasizing the importance of southern boreal fires for regional carbon budgets. Using the southern boreal as an analog, the northern boreal may undergo fundamental shifts in forest structure and carbon dynamics, becoming dominated by stands <70 years old that hold 2-7 kg C/m2 less than current mature northern boreal stands. Our latitudinal approach reinforces previous studies showing that northern boreal stands are at a high risk of holding less carbon under changing disturbance conditions.


Altered airway and cardiac responses in mice lacking G protein-coupled receptor kinase 3.
Contraction and relaxation of airway smooth muscles is mediated, in part, by G protein-coupled receptors (GPCRs) and dysfunction of these receptors has been implicated in asthma. Phosphorylation of GPCRs, by G protein-coupled receptor kinase (GRK), is an important mechanism involved in the dampening of GPCR signaling. To determine whether this mechanism might play a role in airway smooth muscle physiology, we examined the airway pressure time index and heart rate (HR) responses to intravenous administration of the cholinergic agonist methacholine (MCh) in genetically altered mice lacking one copy of GRK2 (GRK2 +/-), homozygous GRK3 knockout (GRK3 -/-), and wild-type littermates. (GRK2 -/- mice die in utero.) GRK3 -/- mice demonstrated a significant enhancement in the airway response to 100 and 250 microgram/kg doses of MCh compared with wild-type and GRK2 +/- mice. GRK3 -/- mice also displayed an enhanced sensitivity of the airway smooth muscle response to MCh. In addition, GRK3 -/- mice displayed an altered HR recovery from MCh-induced bradycardia. Although direct stimulation of cardiac muscarinic receptors measured as vagal stimulation-induced bradycardia was similar in GRK3 -/- and wild-type mice, the baroreflex increase in HR associated with sodium nitroprusside-induced hypotension was significantly greater in GRK3 -/- than wild-type mice. Therefore, these data demonstrate that in the mouse, GRK3 may be involved in modulating the cholinergic response of airway smooth muscle and in regulating the chronotropic component of the baroreceptor reflex.


Temporal shifts in cardiovascular risk factor distribution.
Complementary strategies to shift risk factor population distributions and target high-risk individuals are required to reduce the burden of type 2 diabetes and cardiovascular disease (CVD). To examine secular changes in glucose and CVD risk factors over 20 years during an individual and population-based CVD prevention program in Västerbotten County, Sweden. Population-based health promotion intervention was conducted and annual invitation for individuals turning 40, 50, and 60 years to attend a health assessment, including an oral glucose tolerance test, biochemical measures, and a questionnaire. Data were collected between 1991 and 2010, analyzed in 2012 and available for 120,929 individuals. Linear regression modeling examined age-adjusted differences in CVD risk factor means over time. Data were direct-age-standardized to compare disease prevalence. Between 1991-1995 and 2006-2010, mean age-adjusted cholesterol (men=-0.53, 95% CI=-0.55, -0.50 mmol/L; women=-0.48, 95% CI=-0.50, -0.45 mmol/L) and systolic blood pressure declined (men=-3.06, 95% CI=-3.43, -2.70 mm Hg; women=-5.27, 95% CI=-5.64, -4.90 mm Hg), with corresponding decreases in the age-standardized prevalence of hypertension and hyperlipidemia. Mean age-adjusted 2-hour plasma glucose (men=0.19, 95% CI=0.15, 0.23 mmol/L; women=0.08, 95% CI=0.04, 0.11 mmol/L) and BMI increased (men=1.12, 95% CI=1.04, 1.21; women=0.65, 95% CI=0.55, 0.75), with increases in the age-standardized prevalence of diabetes and obesity. These data demonstrate the potential of combined individual- and population-based approaches to CVD risk factor control and highlight the need for additional strategies addressing hyperglycemia and obesity.


Characterization of the vaccinia virus A35R protein and its role in virulence.
The vaccinia virus A35R gene is highly conserved among poxviruses and encodes a previously uncharacterized hydrophobic acidic protein. Western blotting with anti-A35R peptide antibodies indicated that the protein is expressed early in infection and resolved as a single sharp band of approximately 23 kDa, slightly higher than the 20 kDa predicted from its sequence. The protein band appeared to be the same molecular weight on sodium dodecyl sulfate-polyacrylamide gel electrophoresis, whether expressed in an in vitro transcription/translation system without microsomes or expressed in infected cells, suggesting that it was not glycosylated. A mutant virus with the A35R gene deleted (vA35Delta) formed wild-type-sized plaques on all cell lines tested (human, monkey, mouse, and rabbit); thus, A35R is not required for replication and does not appear to be a host range gene. Although the A35R protein is hydrophobic, it is unlikely to be an integral membrane protein, as it partitioned to the aqueous phase during TX-114 partitioning. The protein could not be detected in virus-infected cell supernatants. A35R localized intracellularly to the virus factories, where the first stages of morphogenesis occur. The vA35Delta mutant formed near-normal levels of the various morphogenic stages of infectious virus particles and supported normal acid-induced fusion of virus-infected cells. Despite normal growth and morphogenesis in vitro, the vA35Delta mutant virus was attenuated in intranasal challenge of mice compared to wild-type and A35R rescue virus. Thus, the intracellular A35R protein plays a role in virulence. The A35R has little homology to any protein outside of poxviruses, suggesting a novel virulence mechanism.


Understanding the "black box" of a health-promotion program: Keys to enable health among older persons aging in the context of migration.
Although the need to make health services more accessible to persons who have migrated has been identified, knowledge about health-promotion programs (HPPs) from the perspective of older persons born abroad is lacking. This study explores the design experiences and content implemented in an adapted version of a group-based HPP developed in a researcher-community partnership. Fourteen persons aged 70-83 years or older who had migrated to Sweden from Finland or the Balkan Peninsula were included. A grounded theory approach guided the data collection and analysis. The findings showed how participants and personnel jointly helped raise awareness. The participants experienced three key processes that could open doors to awareness: enabling community, providing opportunities to understand and be understood, and confirming human values and abilities. Depending on how the HPP content and design are being shaped by the group, the key processes could both inhibit or encourage opening doors to awareness. Therefore, this study provides key insights into how to enable health by deepening the understanding of how the exchange of health-promoting messages is experienced to be facilitated or hindered. This study adds to the scientific knowledge base of how the design and content of HPP may support and recognize the capabilities of persons aging in the context of migration.


Donor-derived thymic-dependent T cells cause chronic graft-versus-host disease.
Chronic graft-versus-host disease (GVHD) is the most common cause of poor long-term outcomes after allogeneic bone marrow transplantation (BMT), but the pathophysiology of chronic GVHD still remains poorly understood. We tested the hypothesis that the impaired thymic negative selection of the recipients will permit the emergence of pathogenic T cells that cause chronic GVHD. Lethally irradiated C3H/HeN (H-2k) recipients were reconstituted with T-cell-depleted bone marrow cells from major histocompatibility complex [MHC] class II-deficient (H2-Ab1-/-) B6 (H-2b) mice. These mice developed diseases that showed all of the clinical and histopathological features of human chronic GVHD. Thymectomy prevented chronic GVHD, thus confirming the causal association of the thymus. CD4+ T cells isolated from chronic GVHD mice were primarily donor reactive, and adoptive transfer of CD4+ T cells generated in these mice caused chronic GVHD in C3H/HeN mice in the presence of B6-derived antigen-presenting cells. Our results demonstrate for the first time that T cells that escape from negative thymic selection could cause chronic GVHD after allogeneic BMT. These results also suggest that self-reactivity of donor T cells plays a role in this chronic GVHD, and improvement in the thymic function may have a potential to decrease chronic GVHD.


Survivorship care for postmenopausal breast cancer patients in Ireland: What do women want?
The aim of this study was to identify the concerns of postmenopausal breast cancer patients in Ireland and inform the development of a survivorship care plan. A qualitative participatory approach was used. Focus group interviews (n = 6) with 51 women were undertaken. Following analysis of the focus group discussions, two nominal group technique (NGT) (consensus workshops) involving representatives (n = 17) from each of the six focus groups were held. Ten key issues were highlighted by women in the focus groups and these were prioritised at the consensus workshops. The most important issues in survivorship care planning prioritised by the women were as follows: meet the same healthcare professional at each review visit; contact number of a named person that you can contact if you have any concerns between review visits; at each review visit, have a physical examination and blood tests and explanation from health care professional outlining if follow up scans needed and if not, why not; information on signs and symptoms of recurrence; advice on diet, exercise, healthy lifestyle and advice on coping and pacing yourself; information and management of side effects of therapy-long and short term. Survivorship care planning for breast cancer is underdeveloped in Ireland. There is a lack of consensus regarding its provision and a lack of structured approach to its implementation. This study demonstrates the role of postmenopausal breast cancer patients' involvement in identifying their needs and reports that continuity of care was their top priority and the need for an adoption of a survivorship care plan was emphasised by participants.


Protective effect of volatile oil, alcoholic and aqueous extracts of Origanum majorana on lead acetate toxicity in mice.
Natural dietary antioxidants are extensively studied for their ability to protect cells from miscellaneous damages. Origanum majorana L., Lamiaceae, is a potent antioxidant. The effect of administration of O. majorana (volatile oil, alcoholic and aqueous extracts) on oral administration of lead acetate in the diet of mice at concentration 0.5% (W/W) for one month were studied by measuring serum alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP), urea and creatinine, histopathological changes of the liver and kidney and genotoxicity including, rate of micronucleus and chromosomal aberrations in bone marrow cells. Mice were treated with the 3 different forms of O. majorana, one month before and maintained with lead acetate administration. The 3 forms of O. majorana induced a significant decrease in serum activities of transaminases (AST & ALT), ALP, urea and creatinine and improved the liver and kidney histology in comparison with lead acetate treated group. Alcoholic extracts of O. majorana significantly reduced the rate of micronucleus, number of aberrant cells and different kinds of chromosomal aberrations. Volatile oil extract significantly reduced the rate of micronucleus and chromosomal fragments. Aqueous extract and volatile oil also of O. majorana significantly reduced number of gaps, ring chromosome and stickiness. It could be concluded that O. majorana plays an important role in ameliorating liver and kidney functions and genotoxicity induced by lead toxicity.


Cervical cancer incidence and mortality in New Mexico's Hispanics, American Indians, and non-Hispanic whites.
High rates of cervical cancer were reported in New Mexico in the early 1970s, with especially high rates for minority women. We examined data collected from 1970 to 1987 for invasive cervical cancer and cervical carcinoma in situ for New Mexico's Hispanic, American Indian, and non-Hispanic white women to determine whether changes had occurred in cervical cancer rates since earlier reports. To further characterize the epidemiology of cervical cancer in New Mexico, we reviewed state vital statistics for cervical cancer deaths occurring between 1958 and 1987. From 1970 to 1987, the incidence for invasive cervical cancer among Hispanic (18.9 per 100,000 person-years) and American Indian women (22.0 per 100,000 person-years) was about double that for non-Hispanic white women (10.3 per 100,000). The incidence in each ethnic group decreased over time for both invasive cancer and carcinoma in situ when the data were examined by 2 time periods (1970 to 1978 and 1979 to 1987). These decreases were most dramatic for invasive cervical cancer. Cervical cancer-related death rates for Hispanics and non-Hispanic whites also decreased from 1958 to 1987. Although our data reflect declines in cervical cancer rates during the study period, further rate decreases, especially for minority women, remain an important public health goal in New Mexico.


Genome-wide mapping and identification of new quantitative trait loci affecting meat production, meat quality, and carcass traits within a Duroc purebred population.
Most QTL detection studies in pigs have been carried out in experimental F(2) populations. However, segregation of a QTL must be confirmed within a purebred population for successful implementation of marker-assisted selection. Previously, QTL for meat quality and carcass traits were detected on SSC 7 in a Duroc purebred population. The objectives of the present study were to carry out a whole-genome QTL analysis (except for SSC 7) for meat production, meat quality, and carcass traits and to confirm the presence of segregating QTL in a Duroc purebred population. One thousand and four Duroc pigs were studied from base to seventh generation; the pigs comprised 1 closed population of a complex multigenerational pedigree such that all individuals were related. The pigs were evaluated for 6 growth traits, 7 body size traits, 8 carcass traits, 2 physiological traits, and 11 meat quality traits, and the number of pigs with phenotypes ranged from 421 to 953. A total of 119 markers were genotyped and then used for QTL analysis. We utilized a pedigree-based, multipoint variance components approach to test for linkage between QTL and the phenotypic values using a maximum likelihood method; the logarithm of odds score and QTL genotypic heritability were estimated. A total of 42 QTL with suggestive linkages and 3 QTL with significant linkages for 26 traits were detected. These included selection traits such as daily BW gain, backfat thickness, loin eye muscle area, and intramuscular fat content as well as correlated traits such as body size and meat quality traits. The present study disclosed QTL affecting growth, body size, and carcass, physiological, and meat quality traits in a Duroc purebred population.


Shaping Magnetite Nanoparticles from First Principles.
Iron oxide magnetic nanoparticles (NPs) are stimuli-responsive materials at the forefront of nanomedicine. Their realistic finite temperature simulations are a formidable challenge for first-principles methods. Here, we use density functional tight binding to open up the required time and length scales and obtain global minimum structures of Fe_{3}O_{4} NPs of realistic size (1400 atoms, 2.5 nm) and of different shapes, which we then refine with hybrid density functional theory methods to accomplish proper electronic and magnetic properties, which have never been accurately described in simulations. On this basis, we develop a general empirical formula and prove its predictive power for the evaluation of the total magnetic moment of Fe_{3}O_{4} NPs. By converting the total magnetic moment into the macroscopic saturation magnetization, we rationalize the experimentally observed dependence with shape. We also reveal interesting reconstruction mechanisms and unexpected patterns of charge ordering.


The impact of serotype-specific vaccination on phylodynamic parameters of Streptococcus pneumoniae and the pneumococcal pan-genome.
In the United States, the introduction of the heptavalent pneumococcal conjugate vaccine (PCV) largely eliminated vaccine serotypes (VT); non-vaccine serotypes (NVT) subsequently increased in carriage and disease. Vaccination also disrupts the composition of the pneumococcal pangenome, which includes mobile genetic elements and polymorphic non-capsular antigens important for virulence, transmission, and pneumococcal ecology. Antigenic proteins are of interest for future vaccines; yet, little is known about how the they are affected by PCV use. To investigate the evolutionary impact of vaccination, we assessed recombination, evolution, and pathogen demographic history of 937 pneumococci collected from 1998-2012 among Navajo and White Mountain Apache Native American communities. We analyzed changes in the pneumococcal pangenome, focusing on metabolic loci and 19 polymorphic protein antigens. We found the impact of PCV on the pneumococcal population could be observed in reduced diversity, a smaller pangenome, and changing frequencies of accessory clusters of orthologous groups (COGs). Post-PCV7, diversity rebounded through clonal expansion of NVT lineages and inferred in-migration of two previously unobserved lineages. Accessory COGs frequencies trended toward pre-PCV7 values with increasing time since vaccine introduction. Contemporary frequencies of protein antigen variants are better predicted by pre-PCV7 values (1998-2000) than the preceding period (2006-2008), suggesting balancing selection may have acted in maintaining variant frequencies in this population. Overall, we present the largest genomic analysis of pneumococcal carriage in the United States to date, which includes a snapshot of a true vaccine-naïve community prior to the introduction of PCV7. These data improve our understanding of pneumococcal evolution and emphasize the need to consider pangenome composition when inferring the impact of vaccination and developing future protein-based pneumococcal vaccines.


Glucose and palmitate uncouple AMPK from autophagy in human aortic endothelial cells.
Dysregulated autophagy and decreased AMP-activated protein kinase (AMPK) activity are each associated with atherogenesis. Atherogenesis is preceded by high circulating concentrations of glucose and fatty acids, yet the mechanism by which these nutrients regulate autophagy in human aortic endothelial cells (HAECs) is not known. Furthermore, whereas AMPK is recognized as an activator of autophagy in cells with few nutrients, its effects on autophagy in nutrient-rich HAECs has not been investigated. We maintained and passaged primary HAECs in media containing 25 mM glucose and incubated them subsequently with 0.4 mM palmitate. These conditions impaired basal autophagy and rendered HAECs more susceptible to apoptosis and adhesion of monocytes, outcomes attenuated by the autophagy activator rapamycin. Glucose and palmitate diminished AMPK activity and phosphorylation of the uncoordinated-51-like kinase 1 (ULK1) at Ser555, an autophagy-activating site targeted by AMPK. 5-Aminoimidazole-4-carboxamide-1-β-d-ribofuranoside (AICAR)-mediated activation of AMPK phosphorylated acetyl-CoA carboxylase, but treatment with AICAR or other AMPK activators (A769662, phenformin) did not restore ULK1 phosphorylation or autophagosome formation. To determine whether palmitate-induced ceramide accumulation contributed to this finding, we overexpressed a ceramide-metabolizing enzyme, acid ceramidase. The increase in acid ceramidase expression ameliorated the effects of excess nutrients on ULK1 phosphorylation, without altering the effects of the AMPK activators. Thus, unlike low nutrient conditions, AMPK becomes uncoupled from autophagy in HAECs in a nutrient-rich environment, such as that found in patients with increased cardiovascular risk. These findings suggest that combinations of AMPK-independent and AMPK-dependent therapies may be more effective alternatives than either therapy alone for treating nutrient-induced cellular dysfunction.


N-aromatic heterocycle adducts of bulky [1,2,4-(Me3C)3C5H2]2Sm: synthesis, structure and solution analysis.
The reactions of the bulky samarocene [1,2,4-(Me3C)3C5H2]2Sm (1) with several N-aromatic heterocycles, namely, pyridine (3), picoline (4), 4-tert-butyl-pyridine (5), isoquinoline (6) and quinoline (7), are reported. All reactions proceed smoothly in pentane in good yield and crystals of all adducts have been obtained in moderate to good yield and analyzed by X-ray crystallography. The X-ray crystal structures are in agreement with Sm(II) coordination adducts in which no electrons are transferred to the N-aromatic heterocycle. Magnetic data were recorded for 1 and for all adducts, 3-7, and reinforce the electronic structure of Sm(II), f(6) simple coordination adducts. (1)H NMR at different temperatures and visible spectroscopy in toluene show that fast exchange of the N-aromatic heterocyclic ligand (L) occurs in solution according to a ML = M + L equilibrium in which the equilibrium constant is dependent on the bulkiness of L. This fast exchange is stopped below a coalescence temperature T(c), at which the three tert-butyl groups of the cyclopentadienyl are differentiated. The associated rotational barriers have been calculated. Cyclic voltammetry of 1 in THF showed a quasi-reversible wave at a reducing potential of -2.10 V versus the ferricinium/ferrocene couple for the Sm(III)-Sm(II) couple and leads to the conclusion that the non-reduction of L lies in the sterics of 1.


Manipulation of rhizosphere bacterial communities to induce suppressive soils.
Naturally occurring disease-suppressive soils have been documented in a variety of cropping systems, and in many instances the biological attributes contributing to suppressiveness have been identified. While these studies have often yielded an understanding of operative mechanisms leading to the suppressive state, significant difficulty has been realized in the transfer of this knowledge into achieving effective field-level disease control. Early efforts focused on the inundative application of individual or mixtures of microbial strains recovered from these systems and known to function in specific soil suppressiveness. However, the introduction of biological agents into non-native soil ecosystems typically yielded inconsistent levels of disease control. Of late, greater emphasis has been placed on manipulation of the cropping system to manage resident beneficial rhizosphere microorganisms as a means to suppress soilborne plant pathogens. One such strategy is the cropping of specific plant species or genotypes or the application of soil amendments with the goal of selectively enhancing disease-suppressive rhizobacteria communities. This approach has been utilized in a system attempting to employ biological elements resident to orchard ecosystems as a means to control the biologically complex phenomenon termed apple replant disease. Cropping of wheat in apple orchard soils prior to re-planting the site to apple provided control of the fungal pathogen Rhizoctonia solani AG-5. Disease control was elicited in a wheat cultivar-specific manner and functioned through transformation of the fluorescent pseudomonad population colonizing the rhizosphere of apple. Wheat cultivars that induced disease suppression enhanced populations of specific fluorescent pseudomonad genotypes with antagonistic activity toward R. solani AG-5, but cultivars that did not elicit a disease-suppressive soil did not modify the antagonistic capacity of this bacterial community. Alternatively, brassicaceae seed meal amendments were utilized to develop soil suppressiveness toward R. solani. Suppression of Rhizoctonia root rot in response to seed meal amendment required the activity of the resident soil microbiota and was associated with elevated populations of Streptomyces spp. recovered from the apple rhizosphere. Application of individual Streptomyces spp. to soil systems provided control of R. solani to a level and in a manner equivalent to that obtained with the seed meal amendment. These and other examples suggest that management of resident plant-beneficial rhizobacteria may be a viable method for control of specific soilborne plant pathogens.


Acquired potential N-glycosylation sites within the tumor-specific immunoglobulin heavy chains of B-cell malignancies.
Among B-cell malignancies, follicular lymphomas (FL) more frequently show acquired, potential N-glycosylation sites (AGS) within tumor-specific immunoglobulin. The aim of this study was to extend this observation and to evaluate the pattern of presentation of AGS within five different forms of B-cell lymphoma. We sequenced the tumor-specific immunoglobulin heavy chain variable region fragment, including complementarity-determining regions 2 and 3, of forty-seven consecutive patients with a B-cell malignancy enrolled in idiotype vaccine clinical trials. This sequencing approach is known to allow the identification of most AGS. We then statistically analyzed differences in presentation pattern, in terms of tumor histology, immunoglobulin isotype, AGS location and amino acid composition. All twenty-four FL cases presented with at least one AGS, whereas the vast majority of four B-cell lymphoma types other than FL did not. The non- FL group of tumors included four cases of Burkitt's lymphoma, six of diffuse large cell lymphoma, seven mantle cell lymphomas and six small lymphocytic lymphomas. Most IgM-bearing follicular lymphoma cases featured their AGS within complementarity-determining region 2, as opposed to those bearing an IgG, which mostly displayed the AGS within complementarity-determining region 3. The vast majority of AGS located within either complementarity-determining region ended with a serine residue, whereas those located within framework regions mostly featured threonine as the last amino acid residue. In our series, all cases of FL had AGS within their tumor-specific immunoglobulin heavy chain variable regions. In contrast, most B-cell malignancies other than FL did not. Further studies are warranted in order to establish the possible meaning of these findings in terms of disease pathogenesis, their diagnostic value in doubtful cases and their potential implications for immunotherapy.


A qualitative study evaluating experiences of a lifestyle intervention in men with prostate cancer undergoing androgen suppression therapy.
The severe iatrogenic hypogonadal state induced by medical castration used for treatment of prostate cancer is associated with adverse effects including fatigue, increased fracture risk, and a decrease in skeletal muscle function, which negatively impact quality of life. We have previously reported beneficial changes in healthy lifestyle behaviors, physical function and fatigue as a result of a novel combined exercise and dietary advice intervention (a lifestyle intervention) in men with prostate cancer on androgen suppression therapy (AST). The aim of this research was to conduct a qualitative evaluation of the lifestyle intervention in these men with advanced prostate cancer receiving androgen suppression therapy (AST). Twelve men with prostate cancer on AST took part in three focus groups in a UK higher education institution following the 12 week intervention. Sessions lasted between 45 and 60 minutes in duration. All discussions were audio-taped and transcribed. A framework analysis approach was applied to the focus group data. An initial coding framework was developed from a priori issues listed in the topic guide and extended and refined following initial familiarization with the focus group transcripts. Line by line indexing of the transcripts was undertaken iteratively to allow for the incorporation of new codes. Coded sections of text were grouped together (charted) into themes and subthemes prior to a further process of comparison and interpretation. None of the participants involved in the trial were provided with information on how lifestyle changes might be beneficial to men with prostate cancer during the course of their standard medical treatment. We present novel findings that this intervention was considered beneficial for reducing anxiety around treatment and fear of disease progression. Men were supportive of the benefits of the intervention over conventional cancer survival discussion group arrangements as it facilitated peer support in addition to physical rehabilitation. The benefits of lifestyle changes in men with prostate cancer are not well appreciated by care providers despite a range of benefits becoming apparent. Strategies to implement exercise and dietary interventions in standard care should be further evaluated. Current Controlled Trials ISRCTN88605738.


Nondestructive readout of a photorefractive hologram by phase-conjugate copying in a one-crystal configuration.
We propose a novel phase-conjugate copying method for nondestructive readout of a volatile photorefractive hologram. In the one-crystal configuration, two holographic memories and a mutually pumped phase conjugator (MPPC) are formed within a single photorefractive crystal, instead of using multiple crystals. Two memories share the same hologram and complement each other in refreshing the hologram. A MPPC suppresses fanning noise and automatically aligns the wavefront of the reference and readout beams. We found the optimum configuration to achieve nondestructive readout from calculations and geometric consideration. In the experiments with a BaTiO(3) crystal, a continuous readout of 20 times longer than the recording time was achieved.


APOBEC mutagenesis is a common process in normal human small intestine.
APOBEC mutational signatures SBS2 and SBS13 are common in many human cancer types. However, there is an incomplete understanding of its stimulus, when it occurs in the progression from normal to cancer cell and the APOBEC enzymes responsible. Here we whole-genome sequenced 342 microdissected normal epithelial crypts from the small intestines of 39 individuals and found that SBS2/SBS13 mutations were present in 17% of crypts, more frequent than most other normal tissues. Crypts with SBS2/SBS13 often had immediate crypt neighbors without SBS2/SBS13, suggesting that the underlying cause of SBS2/SBS13 is cell-intrinsic. APOBEC mutagenesis occurred in an episodic manner throughout the human lifespan, including in young children. APOBEC1 mRNA levels were very high in the small intestine epithelium, but low in the large intestine epithelium and other tissues. The results suggest that the high levels of SBS2/SBS13 in the small intestine are collateral damage from APOBEC1 fulfilling its physiological function of editing APOB mRNA.


Direct evidence that the poly(A) tail of influenza A virus mRNA is synthesized by reiterative copying of a U track in the virion RNA template.
The poly(A) tail of influenza virus mRNA is thought to be synthesized by reiterative copying of the U track near the 5' end of the virion RNA template. This has been widely accepted as a plausible hypothesis, but until now there has been no direct experimental evidence for it. Here, we report such direct evidence based on the fact that (i) replacing the U track with an A track directs synthesis of products with poly(U) tails, both in vitro and in vivo, and (ii) interrupting the U track abolishes polyadenylation in vitro.


Ion channels and transporters in cancer. 3. Ion channels in the tumor cell-microenvironment cross talk.
The traditional view of cancer as a collection of proliferating cells must be reconsidered, and cancer must be viewed as a "tissue" constituted by both transformed cells and a heterogeneous microenvironment, that tumor cells construct and remodel during multistep tumorigenesis. The "tumor microenvironment" (TM) is formed by mesenchymal, endothelial, and immune cells immersed in a network of extracellular matrix (ECM) proteins and soluble factors. The TM strongly contributes to tumor progression, through long distance, cell-to-cell or cell-to-matrix signals, which influence different aspects of tumor cell behavior. Understanding the relationships among the different components of the cancer tissue is crucial to design and develop new therapeutic strategies. Ion channels are emerging as relevant players in the cross talk between tumor cells and their TM. Ion channels are expressed on tumor cells, as well as in the different cellular components of the TM. In all these cells, ion channels are in a strategic position to sense and transmit extracellular signals into the intracellular machinery. Often, this transmission is mediated by integrin adhesion receptors, which can be functional partners of ion channels since they form molecular complexes with the channel protein in the context of the plasma membrane. The same relevant role is exerted by ion transporters, which also contribute to determine two facets of the cancer tissue: hypoxia and the acidic extracellular pH. On the whole, it is conceivable to prospect the targeting of ion channels for new therapeutic strategies aimed at better controlling the malignant progression of the cancer tissue.


The Role of Mineral and Trace Element Supplementation in Exercise and Athletic Performance: A Systematic Review.
Minerals and trace elements (MTEs) are micronutrients involved in hundreds of biological processes. Deficiency in MTEs can negatively affect athletic performance. Approximately 50% of athletes have reported consuming some form of micronutrient supplement; however, there is limited data confirming their efficacy for improving performance. The aim of this study was to systematically review the role of MTEs in exercise and athletic performance. Six electronic databases and grey literature sources (MEDLINE; EMBASE; CINAHL and SportDISCUS; Web of Science and clinicaltrials.gov) were searched, in accordance with PRISMA guidelines. Results: 17,433 articles were identified and 130 experiments from 128 studies were included. Retrieved articles included Iron (n = 29), Calcium (n = 11), Magnesium, (n = 22), Phosphate (n = 17), Zinc (n = 9), Sodium (n = 15), Boron (n = 4), Selenium (n = 5), Chromium (n = 12) and multi-mineral articles (n = 5). No relevant articles were identified for Copper, Manganese, Iodine, Nickel, Fluoride or Cobalt. Only Iron and Magnesium included articles of sufficient quality to be assigned as 'strong'. Currently, there is little evidence to support the use of MTE supplementation to improve physiological markers of athletic performance, with the possible exception of Iron (in particular, biological situations) and Magnesium as these currently have the strongest quality evidence. Regardless, some MTEs may possess the potential to improve athletic performance, but more high quality research is required before support for these MTEs can be given. PROSPERO preregistered (CRD42018090502).


Challenges and opportunities in graduate nursing education by distributed learning in Canada and Brazil.
In this paper, the authors share their experience related to graduate nursing programs offered by distributed learning (DL) in Canada and Brazil. Although degrees offered by DL are often the subject of criticism, the authors' experience has been that learning outcomes have been very good. Nevertheless, a number of challenges and opportunities have been encountered including those associated with flexibility of the program, delivering practice courses at a distance, facilitating interaction, faculty workload and preparation and student support, Newer technologies that may assist in this effort are identified. Despite the challenges encountered, students rate the program highly and ongoing efforts are underway to ensure excellence of such flexible innovative graduate programs in nursing. The authors argue that despite the challenges, DL programs offer high quality graduate education that meets the needs of many nurses.