Proton pump inhibitors and vascular function: A prospective cross-over pilot study.
Proton pump inhibitors (PPIs) are commonly used drugs for the treatment of gastric reflux. Recent retrospective cohorts and large database studies have raised concern that the use of PPIs is associated with increased cardiovascular (CV) risk. However, there is no prospective clinical study evaluating whether the use of PPIs directly causes CV harm. We conducted a controlled, open-label, cross-over pilot study among 21 adults aged 18 and older who are healthy (n=11) or have established clinical cardiovascular disease (n=10). Study subjects were assigned to receive a PPI (Prevacid; 30 mg) or a placebo pill once daily for 4 weeks. After a 2-week washout period, participants were crossed over to receive the alternate treatment for the ensuing 4 weeks. Subjects underwent evaluation of vascular function (by the EndoPAT technique) and had plasma levels of asymmetric dimethylarginine (ADMA, an endogenous inhibitor of endothelial function previously implicated in PPI-mediated risk) measured prior to and after each treatment interval. We observed a marginal inverse correlation between the EndoPAT score and plasma levels of ADMA (r = -0.364). Subjects experienced a greater worsening in plasma ADMA levels while on PPI than on placebo, and this trend was more pronounced amongst those subjects with a history of vascular disease. However, these trends did not reach statistical significance, and PPI use was also not associated with an impairment in flow-mediated vasodilation during the course of this study. In conclusion, in this open-label, cross-over pilot study conducted among healthy subjects and coronary disease patients, PPI use did not significantly influence vascular endothelial function. Larger, long-term and blinded trials are needed to mechanistically explain the correlation between PPI use and adverse clinical outcomes, which has recently been reported in retrospective cohort studies.


How lifespan associated genes modulate aging changes: lessons from analysis of longitudinal data.
The influence of genes on human lifespan is mediated by biological processes that characterize body's functioning. The age trajectories of these processes contain important information about mechanisms linking aging, health, and lifespan. The objective of this paper is to investigate regularities of aging changes in different groups of individuals, including individuals with different genetic background, as well as their connections with health and lifespan. To reach this objective we used longitudinal data on four physiological variables, information about health and lifespan collected in the Framingham Heart Study (FHS), data on longevity alleles detected in earlier study, as well as methods of statistical modeling. We found that phenotypes of exceptional longevity and health are linked to distinct types of changes in physiological indices during aging. We also found that components of aging changes differ in groups of individuals with different genetic background. These results suggest that factors responsible for exceptional longevity and health are not necessary the same, and that postponing aging changes is associated with extreme longevity. The genetic factors which increase lifespan are associated with physiological changes typical of healthy and long-living individuals, smaller mortality risks from cancer and CVD and better estimates of adaptive capacity in statistical modeling. This indicates that extreme longevity and health related traits are likely to be less heterogeneous phenotypes than lifespan, and studying these phenotypes separately from lifespan may provide additional information about mechanisms of human aging and its relation to chronic diseases and lifespan.


Predictors of cognitive impairment in primary age-related tauopathy: an autopsy study.
Primary age-related tauopathy (PART) is a form of Alzheimer-type neurofibrillary degeneration occurring in the absence of amyloid-beta (Aβ) plaques. While PART shares some features with Alzheimer disease (AD), such as progressive accumulation of neurofibrillary tangle pathology in the medial temporal lobe and other brain regions, it does not progress extensively to neocortical regions. Given this restricted pathoanatomical pattern and variable symptomatology, there is a need to reexamine and improve upon how PART is neuropathologically assessed and staged. We performed a retrospective autopsy study in a collection (n = 174) of post-mortem PART brains and used logistic regression to determine the extent to which a set of clinical and neuropathological features predict cognitive impairment. We compared Braak staging, which focuses on hierarchical neuroanatomical progression of AD tau and Aβ pathology, with quantitative assessments of neurofibrillary burden using computer-derived positive pixel counts on digitized whole slide images of sections stained immunohistochemically with antibodies targeting abnormal hyperphosphorylated tau (p-tau) in the entorhinal region and hippocampus. We also assessed other factors affecting cognition, including aging-related tau astrogliopathy (ARTAG) and atrophy. We found no association between Braak stage and cognitive impairment when controlling for age (p = 0.76). In contrast, p-tau burden was significantly correlated with cognitive impairment even when adjusting for age (p = 0.03). The strongest correlate of cognitive impairment was cerebrovascular disease, a well-known risk factor (p < 0.0001), but other features including ARTAG (p = 0.03) and hippocampal atrophy (p = 0.04) were also associated. In contrast, sex, APOE, psychiatric illness, education, argyrophilic grains, and incidental Lewy bodies were not. These findings support the hypothesis that comorbid pathologies contribute to cognitive impairment in subjects with PART. Quantitative approaches beyond Braak staging are critical for advancing our understanding of the extent to which age-related tauopathy changes impact cognitive function.


Plant polyadenylation factors: conservation and variety in the polyadenylation complex in plants.
Polyadenylation, an essential step in eukaryotic gene expression, requires both cis-elements and a plethora of trans-acting polyadenylation factors. The polyadenylation factors are largely conserved across mammals and fungi. The conservation seems also extended to plants based on the analyses of Arabidopsis polyadenylation factors. To extend this observation, we systemically identified the orthologs of yeast and human polyadenylation factors from 10 plant species chosen based on both the availability of their genome sequences and their positions in the evolutionary tree, which render them representatives of different plant lineages. The evolutionary trajectories revealed several interesting features of plant polyadenylation factors. First, the number of genes encoding plant polyadenylation factors was clearly increased from "lower" to "higher" plants. Second, the gene expansion in higher plants was biased to some polyadenylation factors, particularly those involved in RNA binding. Finally, while there are clear commonalities, the differences in the polyadenylation apparatus were obvious across different species, suggesting an ongoing process of evolutionary change. These features lead to a model in which the plant polyadenylation complex consists of a conserved core, which is rather rigid in terms of evolutionary conservation, and a panoply of peripheral subunits, which are less conserved and associated with the core in various combinations, forming a collection of somewhat distinct complex assemblies. The multiple forms of plant polyadenylation complex, together with the diversified polyA signals may explain the intensive alternative polyadenylation (APA) and its regulatory role in biological functions of higher plants.


Clostridium sticklandii, a specialist in amino acid degradation:revisiting its metabolism through its genome sequence.
Clostridium sticklandii belongs to a cluster of non-pathogenic proteolytic clostridia which utilize amino acids as carbon and energy sources. Isolated by T.C. Stadtman in 1954, it has been generally regarded as a "gold mine" for novel biochemical reactions and is used as a model organism for studying metabolic aspects such as the Stickland reaction, coenzyme-B12- and selenium-dependent reactions of amino acids. With the goal of revisiting its carbon, nitrogen, and energy metabolism, and comparing studies with other clostridia, its genome has been sequenced and analyzed. C. sticklandii is one of the best biochemically studied proteolytic clostridial species. Useful additional information has been obtained from the sequencing and annotation of its genome, which is presented in this paper. Besides, experimental procedures reveal that C. sticklandii degrades amino acids in a preferential and sequential way. The organism prefers threonine, arginine, serine, cysteine, proline, and glycine, whereas glutamate, aspartate and alanine are excreted. Energy conservation is primarily obtained by substrate-level phosphorylation in fermentative pathways. The reactions catalyzed by different ferredoxin oxidoreductases and the exergonic NADH-dependent reduction of crotonyl-CoA point to a possible chemiosmotic energy conservation via the Rnf complex. C. sticklandii possesses both the F-type and V-type ATPases. The discovery of an as yet unrecognized selenoprotein in the D-proline reductase operon suggests a more detailed mechanism for NADH-dependent D-proline reduction. A rather unusual metabolic feature is the presence of genes for all the enzymes involved in two different CO2-fixation pathways: C. sticklandii harbours both the glycine synthase/glycine reductase and the Wood-Ljungdahl pathways. This unusual pathway combination has retrospectively been observed in only four other sequenced microorganisms. Analysis of the C. sticklandii genome and additional experimental procedures have improved our understanding of anaerobic amino acid degradation. Several specific metabolic features have been detected, some of which are very unusual for anaerobic fermenting bacteria. Comparative genomics has provided the opportunity to study the lifestyle of pathogenic and non-pathogenic clostridial species as well as to elucidate the difference in metabolic features between clostridia and other anaerobes.


Foot massage and physiological stress in people with dementia: a randomized controlled trial.
The anxiety associated with unfamiliar surroundings, the disorientation and mental confusion, and the social isolation that accompanies dementia can often create increased stress for people living in long-term care settings. Such a response is thought to affect the autonomic nervous system and result in emotional and physical symptoms of distress that may be manifested as agitation. There is the potential for such distress to influence the physiological response and in particular Blood Pressure and Heart Rate. A relaxation intervention such as massage may influence the physiological stress response. This randomized controlled trial aimed to compare the effect of foot massage (FM) versus a control activity (quiet presence, QP) on physiological stress response (i.e., blood pressure [BP] and heart rate [HR]) in people living with moderate-to-severe dementia in long-term-care settings. Fifty-three residents were randomized to intervention (10-minute FM) or control group (QP). While the FM group experienced a greater reduction in HR than the control group, these reductions were not significantly different between groups (p=0.83; see Table 1 ), or across time (p=0.46). Both groups experienced a reduction in systolic BP and diastolic BP, while the mean reduction in systolic BP was greater for those in the FM group. While the findings do not provide strong support for FM, the finding that both conditions allowed the person with dementia to rest in the presence of another human being is of importance in the care of people with dementia. The close presence of another person may in fact promote relaxation and therefore improve BP and HR measures.


High-performance lithium battery anodes using silicon nanowires.
There is great interest in developing rechargeable lithium batteries with higher energy capacity and longer cycle life for applications in portable electronic devices, electric vehicles and implantable medical devices. Silicon is an attractive anode material for lithium batteries because it has a low discharge potential and the highest known theoretical charge capacity (4,200 mAh g(-1); ref. 2). Although this is more than ten times higher than existing graphite anodes and much larger than various nitride and oxide materials, silicon anodes have limited applications because silicon's volume changes by 400% upon insertion and extraction of lithium which results in pulverization and capacity fading. Here, we show that silicon nanowire battery electrodes circumvent these issues as they can accommodate large strain without pulverization, provide good electronic contact and conduction, and display short lithium insertion distances. We achieved the theoretical charge capacity for silicon anodes and maintained a discharge capacity close to 75% of this maximum, with little fading during cycling.


Effects of the dopamine D2 allosteric modulator, PAOPA, on the expression of GRK2, arrestin-3, ERK1/2, and on receptor internalization.
The activity of G protein-coupled receptors (GPCRs) is intricately regulated by a range of intracellular proteins, including G protein-coupled kinases (GRKs) and arrestins. Understanding the effects of ligands on these signaling pathways could provide insights into disease pathophysiologies and treatment. The dopamine D2 receptor is a GPCR strongly implicated in the pathophysiology of a range of neurological and neuropsychiatric disorders, particularly schizophrenia. Previous studies from our lab have shown the preclinical efficacy of a novel allosteric drug, 3(R)-[(2(S)-pyrrolidinylcarbonyl)amino]-2-oxo-1-pyrrolidineacetamide (PAOPA), in attenuating schizophrenia-like behavioural abnormalities in rodent models of the disease. As an allosteric modulator, PAOPA binds to a site on the D2 receptor, which is distinct from the endogenous ligand-binding site, in order to modulate the binding of the D2 receptor ligand, dopamine. The exact signaling pathways affected by this allosteric modulator are currently unknown. The objectives of this study were to decipher the in vivo effects, in rats, of chronic PAOPA administration on D2 receptor regulatory and downstream molecules, including GRK2, arrestin-3 and extracellular receptor kinase (ERK) 1/2. Additionally, an in vitro cellular model was also used to study PAOPA's effects on D2 receptor internalization. Results from western immunoblots showed that chronic PAOPA treatment increased the striatal expression of GRK2 by 41%, arrestin-3 by 34%, phospho-ERK1 by 51% and phospho-ERK2 by 36%. Results also showed that the addition of PAOPA to agonist treatment in cells increased D2 receptor internalization by 33%. This study provides the foundational evidence of putative signaling pathways, and changes in receptor localization, affected by treatment with PAOPA. It improves our understanding on the diverse mechanisms of action of allosteric modulators, while advancing PAOPA's development into a novel drug for the improved treatment of schizophrenia.


Intravenous Immunoglobulin as a Therapeutic Option for Mycoplasma pneumoniae Encephalitis.
To analyze the outcomes of a cohort of children diagnosed with Mycoplasma pneumoniae encephalitis whose treatment regimens included intravenous immunoglobulin (IVIG). A retrospective study was performed at a single center between 2011 and 2016 of children diagnosed with Mycoplasma pneumoniae encephalitis whose acute treatment regimen included IVIG. Details of therapeutic interventions and the clinical course were retrieved from medical records via an institutionally approved protocol. The modified Rankin score was used to quantify outcomes. Four children met inclusion criteria, 3 of whom had prodromal symptoms of infection lasting 5 to 7 days before onset of their neurologic symptoms. One patient presented with neurologic symptoms with no clinical prodrome. The initial treatment regimen included systemic corticosteroids, antibiotics, or both. IVIG was administered for a total dose of 2 g/kg divided over 2 to 4 days to all 4 children. All children showed clinical improvement after IVIG. The 3 children with prodromal symptoms showed immediate and dramatic clinical improvement after IVIG therapy. The immediate response to immunomodulatory therapy in the patients with prodrome suggests that the neurologic syndrome may be caused at least in part by an autoimmune process. The child who did not respond to IVIG had no prodrome, and also had normal electroencephalographic (EEG) and brain magnetic resonance imaging (MRI) findings. These cases suggest that early administration of IVIG should be considered in patients suspected of having Mycoplasma encephalitis, particularly in those who have had prodromal symptoms.


Genetic stability of genome-scale deoptimized RNA virus vaccine candidates under selective pressure.
Recoding viral genomes by numerous synonymous but suboptimal substitutions provides live attenuated vaccine candidates. These vaccine candidates should have a low risk of deattenuation because of the many changes involved. However, their genetic stability under selective pressure is largely unknown. We evaluated phenotypic reversion of deoptimized human respiratory syncytial virus (RSV) vaccine candidates in the context of strong selective pressure. Codon pair deoptimized (CPD) versions of RSV were attenuated and temperature-sensitive. During serial passage at progressively increasing temperature, a CPD RSV containing 2,692 synonymous mutations in 9 of 11 ORFs did not lose temperature sensitivity, remained genetically stable, and was restricted at temperatures of 34 °C/35 °C and above. However, a CPD RSV containing 1,378 synonymous mutations solely in the polymerase L ORF quickly lost substantial attenuation. Comprehensive sequence analysis of virus populations identified many different potentially deattenuating mutations in the L ORF as well as, surprisingly, many appearing in other ORFs. Phenotypic analysis revealed that either of two competing mutations in the virus transcription antitermination factor M2-1, outside of the CPD area, substantially reversed defective transcription of the CPD L gene and substantially restored virus fitness in vitro and in case of one of these two mutations, also in vivo. Paradoxically, the introduction into Min L of one mutation each in the M2-1, N, P, and L proteins resulted in a virus with increased attenuation in vivo but increased immunogenicity. Thus, in addition to providing insights on the adaptability of genome-scale deoptimized RNA viruses, stability studies can yield improved synthetic RNA virus vaccine candidates.


Treatment with adipose tissue-derived mesenchymal stem cells exerts anti-diabetic effects, improves long-term complications, and attenuates inflammation in type 2 diabetic rats.
Long-term diabetes-associated complications are the major causes of morbidity and mortality in individuals with diabetes. These diabetic complications are closely linked to immune system activation along with chronic, non-resolving inflammation, but therapies to directly reverse these complications are still not available. Our previous study demonstrated that mesenchymal stem cells (MSCs) attenuated chronic inflammation in type 2 diabetes mellitus (T2DM), resulting in improved insulin sensitivity and islet function. Therefore, we speculated that MSCs might exert anti-inflammatory effects and promote the reversal of diabetes-induced kidney, liver, lung, heart, and lens diseases in T2DM rats. We induced a long-term T2DM complication rat model by using a combination of a low dose of streptozotocin (STZ) with a high-fat diet (HFD) for 32 weeks. Adipose-derived mesenchymal stem cells (ADSCs) were systemically administered once a week for 24 weeks. Then, we investigated the role of ADSCs in modulating the progress of long-term diabetic complications. Multiple infusions of ADSCs attenuated chronic kidney disease (CKD), nonalcoholic steatohepatitis (NASH), lung fibrosis, and cataracts; improved cardiac function; and lowered serum lipid levels in T2DM rats. Moreover, the levels of inflammatory cytokines in the serum of each animal group revealed that ADSC infusions were able to not only inhibit pro-inflammatory cytokines IL-6, IL-1β, and TNF-α expression but also increase anti-inflammatory cytokine IL-10 systematically. Additionally, MSCs reduced the number of iNOS(+) M1 macrophages and restored the number of CD163(+) M2 macrophages. Multiple intravenous infusions of ADSCs produced significant protective effects against long-term T2DM complications by alleviating inflammation and promoting tissue repair. The present study suggests ADSCs may be a novel, alternative cell therapy for long-term diabetic complications.


Invasion and growth of Plasmodium falciparum is inhibited in fractionated thalassaemic erythrocytes.
Epidemiological and clinical studies have indicated that the thalassaemias may confer protection against malaria. The study reported here investigated this protective effect in vitro, using a new approach which controls for the potential effect of red cell size and age on the virulence of the parasite. A Percoll density gradient method was used to separate alpha- and beta-thalassaemic trait, haemoglobin H and normal red blood cells (RBC) into fractions of different density. Correlations between RBC density, age and size in fractions of all genotypes were established using red cell creatine as an index of cell age. The development of Plasmodium falciparum over 3 erythrocytic cycles (144 h) in whole blood as well as fractionated samples was monitored by slide microscopy and flow cytometry. A significantly reduced rate of parasite invasion and growth was demonstrated in RBC from all thalassaemic genotypes tested. Poor reinvasion rates were noted in the second and third cycles. Increased duration of culture and red cell age also had a greater negative impact on parasite growth in thalassaemic RBC. This poor growth rate was also associated with the arrest of parasite growth at the schizont stage (schizont maturation arrest) and the accumulation of abnormal, trophozoite/schizont stage parasites in the older thalassaemic RBC fractions. These findings suggest a defect in the number and viability of merozoites generated by parasites growing in thalassaemic RBC. Age related factors such as oxidant stress may play a key role in mediating this kind of protective mechanism and deserve further investigation.


Circulating endothelial progenitor cells and endothelial function after chronic Tadalafil treatment in subjects with erectile dysfunction.
We evaluated the effect of a chronic treatment with Tadalafil on progenitor cells (PCs) number and endothelial function in patients with erectile dysfunction (ED) with or without cardiovascular risk factors. Twenty-six subjects with ED and 23 aged matched controls were studied. All subjects underwent blood tests, International Index of Erectile Function (IIEF-5), Nocturnal Penile Tumescence Rigidity Monitoring test (NPTRM), brachial artery flow-mediated dilation (FMD) and PCs count. International index of erectile function, FMD and PC count were re-evaluated in all subjects at the end of Tadalafil and placebo treatment. With respect to controls patients had lower basal FMD (P < 0.05) and basal PCs (P < 0.05). Treatment with Tadalafil determined a significant increase in PCs (P < 0.001) and FMD (P < 0.001) with respect to basal level. Positive correlation was found between basal FMD and PCs (P < 0.05) and between basal FMD and PCs increase after Tadalafil treatment (P < 0.05). Tadalafil promotes a mobilization of PCs and improves endothelial function in ED patients.


RNA and protein localisations of TGF beta 2 in the early mouse embryo suggest an involvement in cardiac development.
We have performed a detailed analysis of the localisations of RNAs for TGF beta 2 and beta 3, and of TGF beta 2 protein in mouse embryos from 6.5 to 9.5 days post coitum, using in situ hybridisation and immunohistochemistry on serial sections, and whole-mount in situ hybridisation to complete embryos. TGF beta 3 RNA was not seen in any of the tissue sections, but very low levels of the RNA were seen by whole-mount in situ hybridisation around the outflow tract of the heart at 8.5 days post coitum. TGF beta 2 RNA is expressed at high levels in all cells with the potential to differentiate into cardiomyocytes. Additionally, the foregut endoderm, juxtaposed to the heart, and the neuroepithelium at the rostral extremity of the foregut, express very high levels of TGF beta 2 RNA, between 8.5 and 9.5 days post coitum. As cardiomyogenesis proceeds, TGF beta 2 RNA levels diminishes within the myocytes, with a concomitant increase in staining for TGF beta 2 protein. TGF beta 2 protein staining of cardiomyocytes persists throughout development and in the adult, in the absence of detectable levels of the corresponding RNA. Superimposed upon this myocardial pattern of expression, there is an upregulation of TGF beta 2 RNA in the myocardium of the outflow tract and atrioventricular canal between 8.5 and 9.5 days post coitum, which returns to low levels by 11.5 days post coitum. The results are discussed in terms of a potential role of TGF beta 2 in controlling cardiomyogenesis and in inductive interactions leading to cardiac cushion tissue formation.


Risk factors for cardiovascular events among Asian patients without pre-existing cardiovascular disease on the renal transplant wait list.
For suitable end-stage renal failure (ESRF) patients, renal transplantation gives better long term survival and quality of life as compared to dialysis. Prior to entry into the renal transplant wait list, potential candidates are screened for the presence of cardiovascular disease. However, the waiting time on the transplant list is long, and interval screening for cardiac fitness for surgery is not well defined. We aim to study the risk factors for the development of a cardiovascular event (CVE) and the time interval from recruitment to onset of a CVE that resulted in their removal from the transplant wait list. A retrospective study of all patients registered under the cadaveric renal transplant waiting list in Singapore General Hospital (SGH) from 16th April 1987 to 31st October 2010. We identified patients who developed a CVE among this cohort. We compared the demographics and clinical characteristics of patients who experienced a CVE versus those who did not. Univariable and multivariable cox regression were performed to investigate the significant variables for the development of a CVE. The time to development of CVE was estimated using Kaplan Meier estimation and log-rank test was used to compare the time to CVE between those with diabetes mellitus and those without. 1265 patients were enrolled in this study. 273 patients dropped out of the wait list due to medical reasons or death, of which 38.8% were due to CVE. The mean and median time duration from recruitment into the waiting list to development of a CVE was 14.42 (95% CI 13.72 to 15.11) and 15.69 (95% CI 13.86 to 17.51) years respectively. For patients with diabetes mellitus, this was 8.22 (95% CI 6.30 to 10.14) and 8.16 (95% CI 4.95 to 11.36) years respectively. Factors associated with an increased risk of developing a CVE included male gender (adjusted HR 2.21, 95% CI 1.43 to 3.41, p<0.001), presence of diabetes mellitus (adjusted HR 5.13, 95% CI 2.85 to 9.24, p<0.001) and patients who were either not working or working part-time as compared to their full-time counterparts (adjusted HR 1.76, 95% CI 1.14 to 2.72, p=0.010). In addition, hazard ratio for CVE significantly increased with advancing age quartile (p<0.001 by log rank test for trend). A significant proportion of patients exited from the renal transplant wait list due to a CVE. Being male, age 37 years old or more, presence of diabetes mellitus and non-working or part-time workers as compared to full-time workers were found to increase the risk of developing a CVE during the wait period for transplantation. The presence of diabetes mellitus significantly shortened the time to development of a CVE.


High-intensity exercise elicits the mobilization of senescent T lymphocytes into the peripheral blood compartment in human subjects.
Clonal expansion of T lymphocytes in response to antigenic stimulation is a fundamental process of adaptive immunity. As a consequence of clonal expansion, some T lymphocytes acquire a senescent phenotype, fail to replicate in response to further antigenic stimulation, and express the killer cell lectin-like receptor G1 (KLRG1) and/or CD57. Physical exercise elicits a mobilization of large numbers of T lymphocytes into the bloodstream from peripheral lymphoid compartments, but the frequency of senescent cells in the mobilized population is not known. Eight male runners (age: 29 +/- 9 yr; maximal O2 uptake 62 +/- 6 ml x kg(-1) x min(-1)) performed an intensive treadmill-running protocol at 80% maximal O2 uptake to volitional exhaustion. Blood lymphocytes isolated before, immediately after, and 1 h after exercise were assessed for cell surface expression of KLRG1, CD57, CD28, CD45RA, CD45RO, CD62L, and lymphocyte subset markers (CD3, CD4, CD8, CD56) by flow cytometry. The percentage of all CD3+ T lymphocytes expressing KLRG1 and CD57 increased with exercise (P < 0.01). The change in T-lymphocyte KLRG1 expression was attributed to both CD4+ and CD8 bright T cells, with the relative change being greater for the CD8 bright population (P < 0.01). Mobilized T-lymphocyte populations expressing KLRG1 and CD57 appeared to extravasate the peripheral blood compartment after 1 h of recovery. In conclusion, T lymphocytes with a senescent phenotype are mobilized and subsequently removed from the bloodstream in response to acute high-intensity exercise. This suggests that T lymphocytes contained within the peripheral lymphoid compartments that are mobilized by exercise are likely to be at a more advanced stage of biological aging and have a reduced capacity for clonal expansion than blood-resident T cells.


Canine subcutaneous mast cell tumour: diagnosis and prognosis.
The aim of this study was to characterize the pathology and clinical outcome of the subcutaneous variant of canine mast cell tumour. Fifty-three cases satisfying the inclusion criteria were selected from the pathology archive of the College of Veterinary Medicine, University of Tennessee. Referring veterinarians provided information on outcome. These dogs had a median age of 9 years (range 3-17 years). After characterizing tumours histologically, nuclear expression of proliferating cell nuclear antigen (PCNA) and Ki67 (MIB-1 clone) was determined immunohistochemically and mast cell origin was confirmed with c-Kit staining. Counts of argyrophilic nucleolar organizer regions (AgNOR) were determined by silver staining. Nuclear labelling was counted in 100 tumour cells. Margins were recorded as incomplete in 66% of dogs, and metastases occurred in 6% of dogs. The estimated minimum mean survival time from date of diagnosis was 1199 days, ranging from 55 to >1780 days. The median scores from immunohistochemical labelling were PCNA 0.05 and Ki67 0.03 per 100 tumour cells. The median score for AgNOR staining was 1.25 per 100 tumour cells. The patterns of c-Kit expression included membranous labelling in 20 tumours, stippled cytoplasmic labelling in 23 tumours and diffuse cytoplasmic labelling in 10 tumours. Age (r=-0.61, P=0.14) and AgNOR score (r=-0.58, P=0.17) had moderate, but non-significant, negative associations with survival. PCNA (r=-0.32, P=0.47), Ki67 (r=-0.22, P=0.64) and c-Kit immunolabelling was not associated with survival. The subcutaneous variant of canine mast cell tumour is distinct in having features of intermediate histological grade and extended mean survival times, suggesting a slightly better long-term prognosis than for higher grade dermal variants. Expression of nuclear proliferation markers is not associated with outcome.


Systemic Oxygen Transport with Rest, Exercise, and Hypoxia: A Comparison of Humans, Rats, and Mice.
The objective of this article is to compare and contrast the known characteristics of the systemic O2 transport of humans, rats, and mice at rest and during exercise in normoxia and hypoxia. This analysis should help understand when rodent O2 transport findings can-and cannot-be applied to human responses to similar conditions. The O2 -transport system was analyzed as composed of four linked conductances: ventilation, alveolo-capillary diffusion, circulatory convection, and tissue capillary-cell diffusion. While the mechanisms of O2 transport are similar in the three species, the quantitative differences are naturally large. There are abundant data on total O2 consumption and on ventilatory and pulmonary diffusive conductances under resting conditions in the three species; however, there is much less available information on pulmonary gas exchange, circulatory O2 convection, and tissue O2 diffusion in mice. The scarcity of data largely derives from the difficulty of obtaining blood samples in these small animals and highlights the need for additional research in this area. In spite of the large quantitative differences in absolute and mass-specific O2 flux, available evidence indicates that resting alveolar and arterial and venous blood PO2 values under normoxia are similar in the three species. Additionally, at least in rats, alveolar and arterial blood PO2 under hypoxia and exercise remain closer to the resting values than those observed in humans. This is achieved by a greater ventilatory response, coupled with a closer value of arterial to alveolar PO2 , suggesting a greater efficacy of gas exchange in the rats. © 2018 American Physiological Society. Compr Physiol 8:1537-1573, 2018.


Utility of patient-activated cardiac event recorders in the detection of cardiac arrhythmias.
Patient-activated event recorders are useful for the diagnosis of arrhythmia in patients with palpitation and presyncope. However, the utility of event recorders in patients suspected of arrhythmia but presenting with other symptoms is not clear. Furthermore, the factors influencing their utility have not been evaluated. Event recorder reports of six hundred and sixty consecutive patients referred due to clinical suspicion of cardiac arrhythmia were reviewed. We divided symptoms into four groups: palpitation, presyncope, chest tightness, and dyspnea. We calculated the diagnostic yield according to patients' symptoms reported on the record, and analyzed the factors affecting the utility of event recorders. The overall diagnostic yield was 64%, and those of palpitation, presyncope, chest tightness, and dyspnea were 66%, 57%, 51% and 60%, respectively. The most common five findings in our patients were sinus rhythm (36%), sinus tachycardia (30%), atrial premature complex (14%), ventricular premature complex (12%), and atrial fibrillation/atrial flutter (7%). The recording duration (8 +/- 4 days) was short in the present study, but the overall diagnostic yield was similar to those of previous ones. Women had lower diagnostic yields than men, especially in atrial flutter-fibrillation. The diagnostic yield was not influenced by age, ordered doctors, or history of cardiovascular disease. CONCLUSIONS; Patient-activated event recorders provided a good diagnostic yield in patients with different presentations of cardiac arrhythmia, and women had lower diagnostic yield in atrial flutter-fibrillation.


Is the presence of 6 or fewer crypt apoptotic bodies sufficient for diagnosis of graft versus host disease? A decade of experience at a single institution.
Histopathology assessment is crucial for the diagnosis of graft versus host disease (GVHD), as the presence of crypt apoptosis is the cardinal criterion required. However, crypt apoptosis is not limited to GVHD; it also occurs in other conditions such as infection, drug reaction, or inflammatory reactions unrelated to GVHD. To better determine whether the presence of 6 or fewer apoptotic bodies is sufficient for the diagnosis of GVHD, we retrospectively reviewed 78 colon biopsies from 66 patients who received either hematopoietic stem cell (HSCT) or cord blood cell transplantation and whose colon biopsies exhibited apoptotic bodies. Among them, 41 cases contained 6 or fewer apoptotic bodies in the colon biopsy. These biopsies were compared with 141 colon biopsy controls that showed no significant pathologic changes as well as 16 colon biopsies with cytomegalovirus colitis from patients without a history of bone marrow transplantation. Among the 41 cases reviewed, 7 patients had coexisting GVHD in other organs (skin or liver). However, gastrointestinal symptoms of at least 4 HSCT patients whose colon biopsies contained 6 or fewer apoptotic bodies completely resolved in the absence of further intervention for GVHD. The discrepancy between pathologic findings and the clinical course may be due to confounding factors, such as infection or medication-induced injury. Our data suggest that identifying 6 or fewer crypt apoptotic bodies in colon biopsies from HSCT patients is worth reporting in order to alert the clinicians of the possibility of GVHD but not sufficient to render a diagnosis on the pathologic grounds alone. The colon biopsies containing 6 or fewer apoptotic bodies represent a heterogenous group. We suggest this group to be classified as indeterminate for GVHD, instead of diagnosing GVHD outright. Synthesis of all clinical, endoscopic, and pathologic information, including the status of infection, coexisting GVHD involvement in the other organs, and medication, is essential for confirmation of the diagnosis of GVHD.


Understory Vegetation Change Following Woodland Reduction Varies by Plant Community Type and Seeding Status: A Region-Wide Assessment of Ecological Benefits and Risks.
Woodland encroachment is a global issue linked to diminished ecosystem services, prompting the need for restoration efforts. However, restoration outcomes can be highly variable, making it difficult to interpret the ecological benefits and risks associated with woodland-reduction treatments within semiarid ecosystems. We addressed this uncertainty by assessing the magnitude and direction of vegetation change over a 15-year period at 129 sagebrush (Artemisia spp.) sites following pinyon (Pinus spp.) and juniper (Juniperus spp.) (P-J) reduction. Pretreatment vegetation indicated strong negative relationships between P-J cover and the abundance of understory plants (i.e., perennial grass and sagebrush cover) in most situations and all three components differed significantly among planned treatment types. Thus, to avoid confounding pretreatment vegetation and treatment type, we quantified overall treatment effects and tested whether distinct response patterns would be present among three dominant plant community types that vary in edaphic properties and occur within distinct temperature/precipitation regimes using meta-analysis (effect size = lnRR = ln[posttreatment cover/pretreatment cover]). We also quantified how restoration seedings contributed to overall changes in key understory vegetation components. Meta-analyses indicated that while P-J reduction caused significant positive overall effects on all shrub and herbaceous components (including invasive cheatgrass [Bromus tectorum] and exotic annual forbs), responses were contingent on treatment- and plant community-type combinations. Restoration seedings also had strong positive effects on understory vegetation by augmenting changes in perennial grass and perennial forb components, which similarly varied by plant community type. Collectively, our results identified specific situations where broad-scale efforts to reverse woodland encroachment substantially met short-term management goals of restoring valuable ecosystem services and where P-J reduction disposed certain plant community types to ecological risks, such as increasing the probability of native species displacement and stimulating an annual grass-fire cycle. Resource managers should carefully weigh these benefits and risks and incorporate additional, appropriate treatments and/or conservation measures for the unique preconditions of a given plant community in order to minimize exotic species responses and/or enhance desirable outcomes.


Potential detection of cancer with fluorinated silicon nanoparticles in 19F MR and fluorescence imaging.
Fluorescence is widely used for cell imaging due to its high sensitivity and rich color choices but limited for in vivo imaging because of its low light penetration. Meanwhile, magnetic resonance imaging (MRI) is widely applied for in vivo diagnosis but not suitable for cell imaging because of its low resolution. As a result of rare background in living organisms, 19F-MRI stands out in several fields of clinical application. Herein, we report a one-pot microwave synthesis of fluorinated silicon nanoparticles (19FSiNPs), for detection of cancer cells and tumors. Based on the quantum effects of the nano-sized nanoparticles, 19FSiNPs can act as a label free dye for ultracontrast cell fluorescence imaging. Our experiments demonstrated that these nanoprobes significantly enhanced in vivo19F/1H MRI contrast in rats with non-small cell lung tumors. Moreover, the resulting 19FSiNPs exhibited high water dispersibility and excellent biocompatibility, which make them promising for both cell imaging and in vivo imaging applications.


Promoting participation in cardiac rehabilitation: patient choices and experiences.
Cardiac rehabilitation can be an effective means for the secondary prevention of coronary heart disease, but a majority of eligible individuals fail to attend or drop out prematurely. Little research has examined patients' decisions about attendance. This paper reports a study examining patients' beliefs and decision-making about cardiac rehabilitation attendance. A purposive sample of patients from a mixed urban-rural region of Scotland was studied in 2001 using focus groups. Those who were eligible for a standardized 12-week cardiac rehabilitation programme were compared, with separate focus groups held for individuals with high attendance (>60% attendance; n = 27), high rates of attrition (<60% attendance; n = 9) and non-attendance (0% attendance; n = 8). A total of 44 patients (33 men; 11 women) took part in eight focus groups. Participants from all groups held sophisticated and cohesive frameworks of beliefs that influenced their attendance decisions. These beliefs related to the self, coronary heart disease, cardiac rehabilitation, other attending patients, and health professionals' knowledge base. An enduring embarrassment about group or public exercise also influenced attendance. Those who attended reported increased faith in their bodies, a heightened sense of fitness and a willingness to support new patients who attended. Reassurance to ease exercise embarrassment should be given before and during the early stages of programmes, and this could be provided by existing patients. Strategies to promote inclusion should address the inhibiting factors identified in the study, and should present cardiac rehabilitation as a comprehensive programme of activities likely to be of benefit to the individual irrespective of personal characteristics, such as age, sex or exercise capacity.


Latently-infected CD4+ T cells are enriched for HIV-1 Tat variants with impaired transactivation activity.
The ability of HIV to establish latent infection in CD4+ lymphocytes represents a major barrier to the eradication of HIV. It is not clear what mechanisms favor latent over productive infection, but prior studies have suggested a role for the viral transcription factor Tat or its RNA target, TAR. Using samples from five individuals who were started on ART within 6 months of infection and achieved a viral load <50 (suppressed), we isolated one- and two-exon tat RNA from HIV propagated ex vivo from baseline plasma and from co-cultures of CD4+ T cells obtained at baseline and suppressed time points. Compared to virus from the baseline plasma (mostly from productively-infected CD4+ T cells), virus from the baseline and suppressed co-cultures (mostly from latently-infected cells) had more Tat variants with impaired transactivation activity. These findings suggest that impaired activity in the Tat-TAR axis may contribute to the establishment of latent infection in CD4+ T cells.


Splice variant transcripts of the anterior gradient 2 gene as a marker of prostate cancer.
Anterior gradient 2 (AGR2) is a gene predominantly expressed in mucus-secreting tissues or in endocrine cells. Its expression is drastically increased in tumors including prostate cancer. Here we investigated whether AGR2 transcript levels can be used as a biomarker to detect prostate cancer (PCa). Using a PCR-based approach, we could show that in addition to the wild-type (AGRwt long and short) transcripts, five other AGR2 splice variants (SV) (referred to as AGR2 SV-C, -E, -F, -G and -H) were present in cancer cell lines. In tissue biopsies, SV-H and AGR2wt (short) distinguished between benign and PCa (p ≤ 0.05 n = 32). In urine exosomes, AGR2 SV-G and SV-H outperformed serum PSA. Receiver operating characteristic (ROC) curves showed the highest discriminatory power of SV-G and SV-H in predicting PCa. AGR2 SV-G and SV-H are potential diagnostic biomarkers for the non-invasive detection of PCa using urine exosomes.


Cardiovascular risk factors in patients with chronic plaque psoriasis: a case-control study on the Brasov County population.
Many studies have suggested that cardiovascular risk factors seem to be more common in patients with psoriasis than in the general population. In this study we aimed to determine the prevalence of cardiovascular risk factors in patients with chronic plaque psoriasis depending on the severity of disease. We conducted a prospective study in Braşov County (Romania) including 142 patients with chronic plaque psoriasis and disease duration of at least six months and 167 controls without psoriasis. The severity of psoriasis was assessed using the psoriasis area and severity index (PASI) score. Along with a thorough medical history and physical examination, serum lipid profile and fasting plasma glucose tests were carried out. The 10-year Framingham risk score (FRS) for general cardiovascular disease, which includes age, gender, total cholesterol, HDL-cholesterol, systolic blood pressure, smoking status, and diabetes mellitus, was applied. The severity of chronic plaque psoriasis was mild in 32 patients (22.53%) and moderate to severe in 110 patients (77.47%). We found a significant higher prevalence of metabolic syndrome in the patient group compared to controls. Individual components of metabolic syndrome like waist circumference, elevated triglycerides, reduced HDL-C, impaired fasting plasma glucose, and arterial hypertension were also more prevalent in patients than in controls. Mean triglycerides, total cholesterol, LDL-cholesterol and HDL-cholesterol levels were significantly raised in patients with psoriasis when compared to controls. The 10-year FRS was significantly higher in patients with psoriasis than in controls (8.36±5.75 vs. 6.61±4.13; P<0.001). FRS was higher in men (P=0.012) and in patients older than 50 years (P=0.008). According to the severity of psoriasis, FRS increases significantly from mild to moderate-to-severe psoriasis (6.82±4.48 to 8.8±6.71; P=0.003). Psoriasis, and especially moderate to severe psoriasis, seems to represent a risk factor for cardiovascular disease. Patients with psoriasis should be risk-assessed for cardiovascular diseases, and comorbidities should be actively managed.


Neuronify: An Educational Simulator for Neural Circuits.
Educational software (apps) can improve science education by providing an interactive way of learning about complicated topics that are hard to explain with text and static illustrations. However, few educational apps are available for simulation of neural networks. Here, we describe an educational app, Neuronify, allowing the user to easily create and explore neural networks in a plug-and-play simulation environment. The user can pick network elements with adjustable parameters from a menu, i.e., synaptically connected neurons modelled as integrate-and-fire neurons and various stimulators (current sources, spike generators, visual, and touch) and recording devices (voltmeter, spike detector, and loudspeaker). We aim to provide a low entry point to simulation-based neuroscience by allowing students with no programming experience to create and simulate neural networks. To facilitate the use of Neuronify in teaching, a set of premade common network motifs is provided, performing functions such as input summation, gain control by inhibition, and detection of direction of stimulus movement. Neuronify is developed in C++ and QML using the cross-platform application framework Qt and runs on smart phones (Android, iOS) and tablet computers as well personal computers (Windows, Mac, Linux).


Interferon Lambda-3 rs12979860 Variants and Response to Pegylated Interferon in Chronic Hepatitis-C Genotype-3.
To assess the role of single nucleotide polymorphisms (SNPs) near the interferon lambda-3 (IFNλ3) (formal IL-28B) gene rs12979860 in predicting sustained virologic response (SVR) in hepatitis-C virus genotype-3 (HCV-3). Descriptive, analytical study. Department of Medicine, The Aga Khan University Hospital, Karachi, from July 2012 to June 2014. Patients with HCV-3 were classified as sustained virologic response (SVR), relapsers and non-responders. SNPrs12979860 was determined by PCR-RFLPprotocol. Differences between categorical variables were assessed by chi-square or Fisher's exact test, while those between continuous variables were evaluated using the Mann-Whitney U-test. Binary logistic regression analysis by forward conditional method was performed by using significant variables with p-values less than 0.05 as the criteria for model inclusion. Out of 115 patients, rs12979860 genotype-CC, CT, TTwas found in 37 (32.2%), 70 (60.9%), and 8 (7%) patients. 72 patients were male with median age of 45 years. Cirrhosis was present in 32 patients. Patients with response failures (no response and relapse, n=36 and 29, respectively) had higher baseline gamma glutamyl transferase (GGT) level (p < 0.001), higher alanine aminotransferase (p=0.027) and cirrhosis (p=0.001) than patients with SVR. Genotype-CC was present in 16/65 in response failures compared to 21/50 who achieved SVR (p=0.048). Rapid virologic response (RVR) (p < 0.001), low GGT(p=0.001) and absence of cirrhosis (p=0.039) were the independent predictive factors for SVR. In patients who could not achieve RVR and in patients with cirrhosis, SVR was seen more in with genotype-CC (p=0.007 and 0.038). In patients infected with HCV-3, IFNλ3 rs12979860, SNPhas less impact on SVR.


Comparison of three different enrichment strategies for serum low molecular weight protein identification using shotgun proteomics approach.
Serum low-molecular weight (LMW) proteins potentially contain useful biological information and their identification can be used to discover novel potential biomarkers. Given the high complexity of serum samples, in the last years several different prefractionation and enrichment strategies have been developed. In this study three different methods, i.e. hydrogel nanoparticles, Proteominer(®) peptide ligand affinity beads and Sartorius Vivaspin(®) centrifugal ultrafiltration device, were compared and evaluated in order to select the best strategy for the enrichment and prefractionation of LMW proteins. A shotgun proteomics approach was adopted, with in-solution proteolytic digestion of the whole protein mixture and determination of the resulting peptides by nanoHPLC coupled with a high-resolution Orbitrap LTQ-XL mass spectrometer. Data analysis, focusing on the LMW proteome (MW ≤ 40 kDa), has shown that the hydrogel nanoparticles performed better in enriching the LMW protein profiles, with 115 proteins identified against 93 and 95 for Proteominer(®) beads and Sartorius Vivaspin(®) device, respectively.


Embryonic craniofacial bone volume and bone mineral density in Fgfr2(+/P253R) and nonmutant mice.
Quantifying multiple phenotypic aspects of individual craniofacial bones across early osteogenesis illustrates differences in typical bone growth and maturation and provides a basis for understanding the localized and overall influence of mutations associated with disease. We quantify the typical pattern of bone growth and maturation during early craniofacial osteogenesis and determine how this pattern is modified in Fgfr2(+/P253R) Apert syndrome mice. Early differences in typical relative bone density increase are noted between intramembranous and endochondral bones, with endochondral bones normally maturing more quickly during the prenatal period. Several craniofacial bones, including the facial bones of Fgfr2(+/P253R) mice, display lower volumes during the earliest days of osteogenesis and lower relative densities until the perinatal period relative to unaffected littermates. Estimates of bone volume and linear measures describing morphology do not necessarily covary, highlighting the value of quantifying multiple facets of gross osteological phenotypes when exploring the influence of a disease causing mutation. Differences in mechanisms of osteogenesis likely underlie differences in intramembranous and endochondral relative density increase. The influence of the FGFR2 P253R mutation on bone volume changes across the prenatal period and again after birth, while its influence on relative bone density is more stable.


Core Stability and Athletic Performance in Male and Female Lacrosse Players.
This study determined the relationship of core stability with power production, agility, and dynamic stability of collegiate lacrosse players and whether core stability is more evident in these performance variables in either males or females. Twenty male and female collegiate lacrosse players (20.3 ± 1.0 years, 173.2 ± 11.8 cm, 72.6 ± 13.0 kg) performed the pro-agility shuttle, the countermovement jump (CMJ), the Star Excursion Balance Test (SEBT), and prone, right lateral, and left lateral planks on two sessions-familiarization and testing. Independent T-tests were used to compare sexes. SPSS 24.0 was used; significance was accepted at p < 0.05. Pearson correlations were used to compare the relationship of core stability to the performance variables in participants. There was a significant relationship found between the prone plank and pro-agility shuttle in all participants (r = -0.50). No significant relationships were found between core stability and performance variables. A significant difference was found in the pro-agility shuttle (p = 0.001) and the CMJ (p = 0.001) but not in core stability or dynamic stability. Agility, power production, and dynamic stability were not related to core stability in neither male or female lacrosse players. There were no significant differences in core stability and dynamic stability between males and females. A significant difference was found in dynamic stability in the SEBT right leg and left leg composite scores between sexes. From these results, it is suggested that core stability may not directly influence the performance variables in collegiate male and female lacrosse players.


The utility of clinical neurophysiological and quantitative sensory testing for trigeminal neuropathy.
This article reviews the utility of neurophysiological recordings and quantitative sensory testing (QST) in providing sensitive, quantitative, and objective tests for the diagnosis and localization of damage to the trigeminal nerve. Electromyography and recordings of the masseter reflex and compound muscle action potential evoked by transcranial magnetic stimulation or direct electrical stimulation of the masseteric nerve can be of value in evaluating the function of a motor neurons supplying the muscles of mastication. Orthodromic recording of the sensory action potential and trigeminal somatosensory-evoked potential recording with the near-nerve stimulation technique are sensitive tools for the investigation of trigeminal sensory Abeta afferents, whereas recordings of polysynaptic trigeminal brainstem reflexes and tactile QST are less sensitive. At late stages of recovery, the blink reflex and masseter inhibitory reflex are often normal, but at earlier stages, the blink reflex recording has good prognostic value, and the presence of a reflex response may confirm continuity of the nerve trunk after partial laceration. Trigeminal small-fiber function (Adelta and C) can be studied with thermal QST of the cool, warm, heat pain, and cold pain detection thresholds or with laser-evoked potential recording. Thermal QST may remain abnormal years after axonal damage and aids in the diagnosis of late sequelae of trigeminal nerve injury. In a study of the diagnostic value of neurography, blink reflex and thermal QST, and various commonly used clinical sensory tests, neurophysiologic tests and thermal QST had better sensitivity (50% to 88% vs 40% to 59%) and negative predictive values (78% to 100% vs 70% to 74%) compared to clinical examination, whereas the specificity (55% to 100%) and positive predictive values (48% to 73%) were similar. At 1 year after trigeminal nerve injury, the risk of a false negative finding with clinical sensory testing was 94%, whereas the combination of nerve conduction recordings and thermal QST increased the diagnostic yield to 100% in patients with long-standing postsurgical sensory alteration. In conclusion, clinical neurophysiological recordings and QST improve the diagnostic accuracy for trigeminal neuropathy.


Two-step K(+)-Na+ and Ag(+)-Na+ ion-exchanged glass waveguides for C-band applications.
A two-step K(+)-Na+ and Ag(+)-Na+ ion-exchange technique is introduced to fabricate single-mode channel waveguides in BK7 glass for the telecom-wavelength region. The dependencies of insertion loss, polarization-dependent loss (PDL), end bending loss of curved waveguides on channel width, diffusion time, and annealing time are investigated. Results show that postannealing is a required process for improving waveguide properties and an optimal annealing time exists. Although relatively narrow mask openings are used in most one-step ion-exchange processes, a wider channel width, to as wide as 10 microm, is preferred for this two-step method. The minimum coupling loss to/from single-mode fiber and the propagation loss is found to be 0.4 dB and 0.3 dB/cm, respectively. For 5-cm-long waveguides the PDL is less than 0.1 dB. For the S-bend structure the cosine curve exhibit apparently a lower bending loss than the double-arc curve.


Psychometric Evaluation of Anxiety, Depression, and Sleep Quality after a Mild Traumatic Brain Injury: A Longitudinal Study.
Introduction. Over 1 million mild traumatic brain injury (mTBI) cases are reported annually worldwide and may result in cognitive, physical, and emotional deterioration; depression; anxiety; and sleep problems. However, studies on long-term mTBI effects are limited. This study included 440 patients, and regular follow-ups of psychological assessments were performed for 2 years. Four questionnaires, including the Pittsburgh sleep quality index (PSQI), Epworth sleepiness scale (ESS), Beck's anxiety inventory (BAI), and Beck's depression inventory (BDI), were used to evaluate sleep problems, daytime sleepiness, anxiety, and depression, respectively. Results show that BAI and BDI scores considerably improved at the 6th-week, 1st-year, and 2nd-year follow-ups compared to baseline, yet these remained significantly different. In addition, anxiety and depression were prominent symptoms in a select subgroup of patients with poor initial evaluations, which improved over the 2 years. However, the ESS and PSQI scores fluctuated only mildly over the same time span. In conclusion, the mTBI patients showed a gradual improvement of anxiety and depression over the 2 years following injury. While anxiety and depression levels for mTBI patients in general did not return to premorbid status, improvements were observed. Sleep disorders persisted and were consistent with initial levels of distress.


One Health and EcoHealth in Ontario: a qualitative study exploring how holistic and integrative approaches are shaping public health practice in Ontario.
There is a growing recognition that many public health issues are complex and can be best understood by examining the relationship between human health and the health of the ecosystems in which people live. Two approaches, One Health and Ecosystem Approaches to Health (EcoHealth), can help us to better understand these intricate and complex connections, and appear to hold great promise for tackling many modern public health dilemmas. Although both One Health and EcoHealth have garnered recognition from numerous health bodies in Canada and abroad, there is still a need to better understand how these approaches are shaping the practice of public health in Ontario.The purpose of this study was to characterize how public health actors in Ontario are influenced by the holistic principles which underlie One Health and EcoHealth, and to identify important lessons from their experiences. Ten semi-structured interviews were conducted with ten participants from the public health sphere in Ontario. Participants encompassed diverse perspectives including infectious disease, food systems, urban agriculture, and environmental health. Interviews were recorded, transcribed and analyzed using qualitative content analysis to identify major themes and patterns. Four major themes emerged from the interviews: the importance of connecting human health with the environment; the role of governance in promoting these ideas; the value of partnerships and collaborations in public health practice; and the challenge of operationalizing holistic approaches to public health. Overall study participants were found to be heavily influenced by concepts couched in EcoHealth and One Health literature, despite a lack of familiarity with these fields. Although One Health and EcoHealth are lesser known approaches in the public health sphere, their holistic and systems-based principles were found to influence the thoughts, values and experiences of public health actors interviewed in this study. This study also highlights the critical role of governance and partnerships in facilitating a holistic approach to health. Further research on governance and partnership models, as well as systems-based organizational working practices, is needed to close the gap between One Health and EcoHealth theory and public health practice.


Epitope targeting of tertiary protein structure enables target-guided synthesis of a potent in-cell inhibitor of botulinum neurotoxin.
Botulinum neurotoxin (BoNT) serotype A is the most lethal known toxin and has an occluded structure, which prevents direct inhibition of its active site before it enters the cytosol. Target-guided synthesis by in situ click chemistry is combined with synthetic epitope targeting to exploit the tertiary structure of the BoNT protein as a landscape for assembling a competitive inhibitor. A substrate-mimicking peptide macrocycle is used as a direct inhibitor of BoNT. An epitope-targeting in situ click screen is utilized to identify a second peptide macrocycle ligand that binds to an epitope that, in the folded BoNT structure, is active-site-adjacent. A second in situ click screen identifies a molecular bridge between the two macrocycles. The resulting divalent inhibitor exhibits an in vitro inhibition constant of 165 pM against the BoNT/A catalytic chain. The inhibitor is carried into cells by the intact holotoxin, and demonstrates protection and rescue of BoNT intoxication in a human neuron model.


The Cannabinoid CB1/CB2 Agonist WIN55212.2 Promotes Oligodendrocyte Differentiation In Vitro and Neuroprotection During the Cuprizone-Induced Central Nervous System Demyelination.
Different types of insults to the CNS lead to axon demyelination. Remyelination occurs when the CNS attempts to recover from myelin loss and requires the activation of oligodendrocyte precursor cells. With the rationale that CB1 receptor is expressed in oligodendrocytes and marijuana consumption alters CNS myelination, we study the effects of the cannabinoid agonist WIN55212.2 in (1) an in vitro model of oligodendrocyte differentiation and (2) the cuprizone model for demyelination. The synthetic cannabinoid agonist WIN55212.2 at 1 μM increased the myelin basic protein mRNA and protein expression in vitro. During cuprizone-induced acute demyelination, the administration of 0.5 mg/kg WIN55212.2 confers more myelinated axons, increased the expression of retinoid X receptor alpha, and declined nogo receptor expression. Controversially, 1 mg/kg of the drug increased the number of demyelinated axons and reduced the expression of nerve growth factor inducible, calreticulin and myelin-related genes coupling specifically with a decrease in 2',3'-cyclic nucleotide 3' phosphodiesterase expression. The cannabinoid agonist WIN55212.2 promotes oligodendrocyte differentiation in vitro. Moreover, 0.5 mg/kg of the drug confers neuroprotection during cuprizone-induced demyelination, while 1 mg/kg aggravates the demyelination process.


HIV and AIDS vulnerability in fishing communities in Mangochi district, Malawi.
The fisheries sector contributes significantly to Malawi's national economy and to the livelihoods of the poor as certain activities in the sector have relatively low barriers to entry. Various studies have shown that the fisheries sector suffers from high HIV prevalence in many low-income countries. In Malawi, HIV prevalence as well as the causes and impact of HIV infections among fisherfolk are yet to be assessed. Participatory action research was conducted in Mangochi District, in the southern part of Lake Malawi, between December 2007 and January 2008, to identify critical HIV-risk points along the value-chain for the Lake Sardine or usipa (Engraulicypris sardella) fishery. Data were collected through interviews with key informants and from focus group discussions at the community, institution, district and market levels. An analysis of vulnerability resulted in the formulation of mechanisms to redress HIV and AIDS prevention and mitigation at each point of vulnerability. The findings show that HIV and AIDS vulnerability in the fishery's market-chain is highest where fish processing and trading influence sexual relationships between fishermen and female fish traders. The period of high usipa catches (December to March) coincides with a period of food shortage and fishing offers income-generating opportunities for many food-insecure households. This increases competition in processing and trading fish, a socio-economic situation that may result in increased instances of transactional sex. The interactions along the usipa fishery's market-chain, from rural to urban settings, also favour the transmission of HIV from areas of high prevalence to areas with otherwise low HIV risk.


Hospital length of stay and clinical outcomes in older STEMI patients after primary PCI: a report from the National Cardiovascular Data Registry.
There has been a decline in hospital length of stay (LOS) after primary percutaneous coronary intervention (PPCI) for ST-segment elevation myocardial infarction (STEMI). The objective of this study was to examine whether shorter LOS is safe for older patients undergoing PPCI for STEMI. The study analyzed patients' characteristics and 30-day outcomes by LOS (short, ≤3 days; medium, 4 to 5 days; long >5 days; where LOS was the discharge date minus the admission date plus 1) among 33,920 patients with STEMI in the linked CathPCI Registry-Centers for Medicare & Medicaid Services dataset who were ≥65 years of age and treated with PPCI from 2004 to 2009. Percents of patients in each category were as follows: 26.9%, 46.3%, and 26.8% for short, medium, and long LOS, respectively. Patients with a long LOS were generally older, female, and had more comorbidities, including cardiogenic shock and multivessel disease. Patients with a short LOS generally had higher ejection fraction and single-vessel disease. There was no significant difference in 30-day all-cause mortality (hazard ratio [HR]: 1.00; 95% confidence interval [CI]: 0.74 to 1.34) or major adverse cardiac events (MACE) (death, readmission for myocardial infarction, unplanned revascularization: HR: 1.03; 95% CI: 0.86 to 1.25) for medium versus short LOS. There was a significant increase in adjusted mortality (HR: 2.30; 95% CI: 1.72 to 3.07) and MACE (HR: 1.75; 95% CI: 1.44 to 2.12) for long versus short LOS. Patients with a very short LOS (1 to 2 days) had significantly increased 30-day mortality and MACE compared with a 3- to 4-day LOS. Patients discharged as early as 48 h after PPCI have outcomes similar to patients who stay in the hospital for 4 to 5 days. Early, but not very early (<48 h), discharge may be safe among selected older patients with STEMI.


Oxidative stress in pancreatic alpha and beta cells as a selection criterion for biocompatible biomaterials.
The clinical success rate of islet transplantation, namely independence from insulin injections, is limited by factors that lead to graft failure, including inflammation, acute ischemia, acute phase response, and insufficient vascularization. The ischemia and insufficient vascularization both lead to high levels of oxidative stress, which are further aggravated by islet encapsulation, inflammation, and undesirable cell-biomaterial interactions. To identify biomaterials that would not further increase damaging oxidative stress levels and that are also suitable for manufacturing a beta cell encapsulation device, we studied five clinically approved polymers for their effect on oxidative stress and islet (alpha and beta cell) function. We found that 300 poly(ethylene oxide terephthalate) 55/poly(butylene terephthalate) 45 (PEOT/PBT300) was more resistant to breakage and more elastic than other biomaterials, which is important for its immunoprotective function. In addition, it did not induce oxidative stress or reduce viability in the MIN6 beta cell line, and even promoted protective endogenous antioxidant expression over 7 days. Importantly, PEOT/PBT300 is one of the biomaterials we studied that did not interfere with insulin secretion in human islets.


Stability of executive functions in first episode psychosis: One year follow up study.
Executive functioning is a multi-dimensional construct covering several sub-processes. The aim of this study was to determine whether executive functions, indexed by a broad range of executive measures remain stable in first episode psychosis (FEP) over time. Eighty-two patients and 107 age and gender matched healthy controls were assessed on five subdomains of executive functioning; working memory, fluency, flexibility, and inhibitory control at baseline and at 1 year follow-up. Results showed that patients performed significantly poorer than controls on all executive measures at both assessment points. In general executive functions remained stable from baseline to follow-up, although both groups improved on measures of inhibitory control and flexibility. In phonemic fluency, controls showed a slight improvement while patients showed a slight decline. Investigation of individual trajectories revealed some fluctuations in both groups over time, but mainly supports the group level findings. The implications of these results are discussed.


The thalamus is more than just a relay.
The lateral geniculate nucleus and pulvinar are examples of two different types of relay: the former is a first order relay, transmitting information from a subcortical source (retina), while the latter is mostly a higher order relay, transmitting information from layer 5 of one cortical area to another cortical area. First and higher order thalamic relays can also be recognized for much of the rest of thalamus, and most of thalamus seems to be comprised of higher order relays. Higher order relays seem especially important to general corticocortical communication, and this challenges and extends the conventional view that such communication is based on direct corticocortical connections.


Delayed Recognition of Disorders of Sex Development (DSD): A Missed Opportunity for Early Diagnosis of Malignant Germ Cell Tumors.
Disorders of sex development (DSD) are defined as a congenital condition in which development of chromosomal, gonadal or anatomical sex is atypical. DSD patients with gonadal dysgenesis or hypovirilization, containing part of the Y chromosome (GBY), have an increased risk for malignant type II germ cell tumors (GCTs: seminomas and nonseminomas). DSD may be diagnosed in newborns (e.g., ambiguous genitalia), or later in life, even at or after puberty. Here we describe three independent male patients with a GCT; two were retrospectively recognized as DSD, based on the histological identification of both carcinoma in situ and gonadoblastoma in a single gonad as the cancer precursor. Hypospadias and cryptorchidism in their history are consistent with this conclusion. The power of recognition of these parameters is demonstrated by the third patient, in which the precursor lesion was diagnosed before progression to invasiveness. Early recognition based on these clinical parameters could have prevented development of (metastatic) cancer, to be treated by systemic therapy. All three patients showed a normal male 46,XY karyotype, without obvious genetic rearrangements by high-resolution whole-genome copy number analysis. These cases demonstrate overlap between DSD and the so-called testicular dysgenesis syndrome (TDS), of significant relevance for identification of individuals at increased risk for development of a malignant GCT.


An update on addressing important peripheral nerve problems: challenges and potential solutions.
From time to time it is thoughtful and productive to review a medical field and reflect upon what are the major issues that need to be addressed and what is being done to do so. This review article is not meant to be all-inclusive but rather focuses on four evolving areas in the field of peripheral nerve disorders and treatments: (1) nerve surgery under ultrasound guidance using a new ultra-minimally invasive thread technique; (2) evolving magnetic resonance imaging (MRI) and ultrasound imaging techniques that are helping to both diagnose and treat a variety of peripheral nerve problems including entrapment neuropathies, traumatic nerve injuries, and masses arising from nerves; (3) promoting recovery after nerve injury using electrical stimulation; and (4) developing animal models to reproduce a severe nerve injury (neurotmetic grade in continuity) that requires a surgical intervention and repair. In each area we first describe the current challenges and then discuss new and emerging techniques and approaches. It is our hope that this article will bring added attention and resources to help better address peripheral nerve problems that remain a challenge for both patients and physicians.


Effects of pulsed 2.856 GHz microwave exposure on BM-MSCs isolated from C57BL/6 mice.
The increasing use of microwave devices over recent years has meant the bioeffects of microwave exposure have been widely investigated and reported. However the exact biological fate of bone marrow MSCs (BM-MSCs) after microwave radiation remains unknown. In this study, the potential cytotoxicity on MSC proliferation, apoptosis, cell cycle, and in vitro differentiation were assayed following 2.856 GHz microwave exposure at a specific absorption rate (SAR) of 4 W/kg. Importantly, our findings indicated no significant changes in cell viability, cell division and apoptosis after microwave treatment. Furthermore, we detected no significant effects on the differentiation ability of these cells in vitro, with the exception of reduction in mRNA expression levels of osteopontin (OPN) and osteocalcin (OCN). These findings suggest that microwave treatment at a SAR of 4 W/kg has undefined adverse effects on BM-MSCs. However, the reduced-expression of proteins related to osteogenic differentiation suggests that microwave can the influence at the mRNA expression genetic level.


Diversification and the rate of molecular evolution: no evidence of a link in mammals.
Recent research has indicated a positive association between rates of molecular evolution and diversification in a number of taxa. However debate continues concerning the universality and cause of this relationship. Here, we present the first systematic investigation of this relationship within the mammals. We use phylogenetically independent sister-pair comparisons to test for a relationship between substitution rates and clade size at a number of taxonomic levels. Total, non-synonymous and synonymous substitution rates were estimated from mitochondrial and nuclear DNA sequences. We found no evidence for an association between clade size and substitution rates in mammals, for either the nuclear or the mitochondrial sequences. We found significant associations between body size and substitution rates, as previously reported. Our results present a contrast to previous research, which has reported significant positive associations between substitution rates and diversification for birds, angiosperms and reptiles. There are three possible reasons for the differences between the observed results in mammals versus other clades. First, there may be no link between substitution rates and diversification in mammals. Second, this link may exist, but may be much weaker in mammals than in other clades. Third, the link between substitution rates and diversification may exist in mammals, but may be confounded by other variables.


The intervention of robot caregivers and the cultivation of children's capability to play.
In this article, the authors examine whether and how robot caregivers can contribute to the welfare of children with various cognitive and physical impairments by expanding recreational opportunities for these children. The capabilities approach is used as a basis for informing the relevant discussion. Though important in its own right, having the opportunity to play is essential to the development of other capabilities central to human flourishing. Drawing from empirical studies, the authors show that the use of various types of robots has already helped some children with impairments. Recognizing the potential ethical pitfalls of robot caregiver intervention, however, the authors examine these concerns and conclude that an appropriately designed robot caregiver has the potential to contribute positively to the development of the capability to play while also enhancing the ability of human caregivers to understand and interact with care recipients.


Barriers to Point-of-Care Testing in India: Results from Qualitative Research across Different Settings, Users and Major Diseases.
Successful point-of-care testing, namely ensuring the completion of the test and treat cycle in the same encounter, has immense potential to reduce diagnostic and treatment delays, and impact patient outcomes. However, having rapid tests is not enough, as many barriers may prevent their successful implementation in point-of-care testing programs. Qualitative research on diagnostic practices may help identify such barriers across different points of care in health systems. In this exploratory qualitative study, we conducted 78 semi-structured interviews and 13 focus group discussions in an urban and rural area of Karnataka, India, with healthcare providers (doctors, nurses, specialists, traditional healers, and informal providers), patients, community health workers, test manufacturers, laboratory technicians, program managers and policy-makers. Participants were purposively sampled to represent settings of hospitals, peripheral labs, clinics, communities and homes, in both the public and private sectors. In the Indian context, the onus is on the patient to ensure successful point-of-care testing across homes, clinics, labs and hospitals, amidst uncoordinated providers with divergent and often competing practices, in settings lacking material, money and human resources. We identified three overarching themes affecting point-of-care testing: the main theme is 'relationships' among providers and between providers and patients, influenced by the cross-cutting theme of 'infrastructure'. Challenges with both result in 'modified practices' often favouring empirical (symptomatic) treatment over treatment guided by testing. Even if tests can be conducted on the spot and infrastructure challenges have been resolved, relationships among providers and between patients and providers are crucial for successful point-of-care testing. Furthermore, these barriers do not act in isolation, but are interlinked and need to be examined as such. Also, a test alone has only limited power to overcome those difficulties. Test developers, policy-makers, healthcare providers and funders need to use these insights in overcoming barriers to point-of-care testing programs.


Injection drug users trained by overdose prevention programs: responses to witnessed overdoses.
In response to the growing public health problem of drug overdose, community-based organizations have initiated overdose prevention programs (OPPs), which distribute naloxone, an opioid antagonist, and teach overdose response techniques. Injection drug users (IDUs) have been targeted for this intervention due to their high risk for drug overdose. Limited research attention has focused on factors that may inhibit or prevent IDUs who have been trained by OPPs to undertake recommended response techniques when responding to a drug overdose. IDUs (n = 30) trained by two OPPs in Los Angeles were interviewed in 2010-2011 about responses to their most recently witnessed drug overdose using an instrument containing both open and closed-ended questions. Among the 30 witnessed overdose events, the victim recovered in 29 cases while the outcome was unknown in one case. Participants responded to overdoses using a variety of techniques taught by OPPs. Injecting the victim with naloxone was the most commonly recommended response while other recommended responses included stimulating the victim with knuckles, calling 911, and giving rescue breathing. Barriers preventing participants from employing recommended response techniques in certain circumstances included prior successes using folk remedies to revive a victim, concerns over attracting police to the scene, and issues surrounding access to or use of naloxone. Practical solutions, such as developing booster sessions to augment OPPs, are encouraged to increase the likelihood that trained participants respond to a drug overdose with the full range of recommended techniques.


Toluene in sewage and sludge in wastewater treatment plants.
Toluene is a compound that often occurs in municipal wastewater ranging from detectable levels up to 237 μg/L. Before the year 2000, the presence of the aromatic hydrocarbons was assigned only to external sources. The Enhanced Biological Nutrients Removal Processes (EBNRP) work according to many different schemes and technologies. For high-efficiency biological denitrification and dephosphatation processes, the presence of volatile fatty acids (VFAs) in sewage is required. VFAs are the main product of organic matter hydrolysis from sewage sludge. However, no attention has been given to other products of the process. It has been found that in parallel to VFA production, toluene formation occurred. The formation of toluene in municipal anaerobic sludge digestion processes was investigated. Experiments were performed on a laboratory scale using sludge from primary and secondary settling tanks of municipal treatment plants. The concentration of toluene in the digested sludge from primary settling tanks was found to be about 42,000 μg/L. The digested sludge supernatant liquor returned to the biological dephosphatation and denitrification processes for sewage enrichment can contain up to 16,500 μg/L of toluene.


Axonal protein synthesis and degradation are necessary for efficient growth cone regeneration.
Axonal regeneration can occur within hours of injury, the first step being the formation of a new growth cone. For sensory and retinal axons, regenerative ability in vivo correlates with the potential to form a new growth cone after axotomy in vitro. We show that this ability to regenerate a new growth cone depends on local protein synthesis and degradation within the axon. Axotomy in vitro leads to a fourfold to sixfold increase in 3H-leucine incorporation in both neurones and axons, starting within 10 min and peaking 1 h after axotomy. Application of protein synthesis inhibitors (cycloheximide and anisomycin) to cut axons, including axons whose cell bodies were removed, or proteasome inhibitors (lactacystin and N-acetyl-Nor-Leu-Leu-Al) all result in a reduction in the proportion of transected axons able to reform growth cones. Similar inhibition of growth cone formation was observed on addition of target of rapamycin (TOR), p38 MAPK (mitogen-activated protein kinase), and caspase-3 inhibitors. Comparing retinal and sensory axons of different developmental stages, levels of ribosomal protein P0 and phosphorylated translation initiation factor are high in sensory axons, lower in embryonic axons, and absent in adult retinal axons. Conditioning lesions, which increase the regenerative ability of sensory axons, lead to increases in intra-axonal protein synthetic and degradative machinery both in vitro and in vivo. Collectively, these findings suggest that local protein synthesis and degradation, controlled by various TOR-, p38 MAPK-, and caspase-dependent pathways, underlie growth cone initiation after axotomy.


Prospective randomized trial of ligasure versus harmonic hemostasis technique in thyroidectomy.
Two surgical devices have become popular in thyroid surgery: a bipolar energy sealing system (B) and ultrasonic coagulation (UC). Retrospective and prospective studies have demonstrated that the use of these surgical devices for thyroidectomy compared with conventional thyroidectomy (clamp-and-tie) techniques reduces operative time and cost. We conducted a prospective randomized clinical trial to determine if there is any difference in operative time and cost between B and UC. A single-blinded prospective randomized controlled trial was conducted at a tertiary referral center. A total of 90 patients who required a thyroidectomy for thyroid cancer, thyroid nodules, or hyperthyroidism were randomized to either B or UC during thyroidectomy. The operative time and cost of thyroidectomy were compared between the two groups. There was no statistically significant difference in patient age, gender, body mass index, indication for thyroidectomy and thyroid gland weight between the two groups. There was no statistically significant difference in operating room cost or total cost for thyroidectomy between the B and UC groups. There was also no statistically significant difference in the operative time between the B and UC groups (187.6 vs. 184.2 min, P = 0.48) or in postoperative complication rates. The only statistically significant difference in total cost was between surgeons independent of the device used (P < 0.01). In thyroid surgery, total cost and operative time were similar between the two surgical devices used.


Randomized comparison of combination chemotherapy with etoposide, bleomycin, and either high-dose or standard-dose cisplatin in children and adolescents with high-risk malignant germ cell tumors: a pediatric intergroup study--Pediatric Oncology Group 9049 and Children's Cancer Group 8882.
To determine in a randomized comparison whether combination chemotherapy with high-dose cisplatin (HDPEB) improves the event-free (EFS) and overall (OS) survival of children and adolescents with high-risk malignant germ cell tumors (MGCT) as compared with standard-dose cisplatin (PEB) and to compare the regimens' toxicity. Between March 1990 and February 1996, 299 eligible patients with stage III and IV gonadal and extragonadal (all stages) MGCT were enrolled onto this Pediatric Oncology Group and Children's Cancer Group study. Chemotherapy included bleomycin 15 units/m(2) on day 1, etoposide 100 mg/m(2) on days 1 through 5, and either high-dose cisplatin 40 mg/m(2) on days 1 through 5 (HDPEB; n = 149) or standard-dose cisplatin 20 mg/m(2) on days 1 through 5 (PEB; n = 150). Patients were evaluated after four cycles of therapy, and those with residual disease underwent surgery. Those with malignant disease in resected specimen received two additional cycles of their assigned regimen. One hundred thirty-four eligible patients with advanced testicular (n = 60) or ovarian (n = 74) tumors and 165 with stage I to IV extragonadal tumors were enrolled. HDPEB treatment resulted in significantly improved 6-year EFS rate +/- SE (89.6% +/- 3.6% v 80.5% +/- 4.8% for PEB; P =.0284). There was no significant difference in OS (HDPEB 91.7% +/- 3.3% v PEB 86.0% +/- 4.1%). Tumor-related deaths were more common after PEB (14 deaths v two deaths). Toxic deaths were more common with HDPEB (six deaths v one death). Other treatment-related toxicities were more common with HDPEB. Combination chemotherapy with HDPEB significantly improves EFS for children with high-risk MGCT. The OS is similar in both regimens, and the significant toxicity associated with HDPEB limits its use.


Cellular uptake mechanism for oligonucleotides: involvement of endocytosis in the uptake of phosphodiester oligonucleotides by a human colorectal adenocarcinoma cell line, HCT-15.
We used a 20-mer antisense oligonucleotide against human MDR1 mRNA to study the mechanism of uptake of oligonucleotides by a human colorectal adenocarcinoma cell line, HCT-15.5'-end-32P-labeled oligonucleotides were mainly used. The uptake of phosphodiester oligonucleotides (D-oligo) by HCT-15 cells increased with time, but the uptake of phosphorothioate oligonucleotides (S-oligo) showed minimal increases with time. HeLa cells showed a 4-fold greater D-oligo uptake than did HCT-15 cells, and other cell lines exhibited a 25-fold lower uptake than did HCT-15 cells. On the other hand, S-oligo uptake varied only severalfold among the cell lines used. Doligo up-take consisted of a saturable component (Michaelis constant K(m) = 0.16 microM and maximal uptake rate Vmax = 4.8 pmol/min/mg for HCT-15 cells and K(m) = 0.21 microM and Vmax = 35.6 pmol/min/mg for HeLa cells), i.e., the difference between the cell types was mainly in Vmax. The uptake of labeled D-oligo was inhibited by unlabeled S-oligo at a much lower concentration (Ki = 0.01 microM) than by unlabeled D-oligo (K(m) = 0.16 microM), although the uptake rate of D-oligo was greater than that of S-oligo. The D-oligo uptake was highly temperature dependent. Of the sulfhydryl-modifying reagents (p-chloromercuriphenyl-sulfonic acid and p-chloromercuribenzoic acid), only p-chloromercuribenzoic acid inhibited D-oligo uptake, suggesting the involvement of a protein whose sulfhydryl residues inside the cell are critical for D-oligo uptake. Chloroquine and monensin, which are known to alter the intracellular fate of ligands and receptors, did not affect D-oligo uptake, but phenylarsine oxide, an inhibitor of internalization processes, inhibited uptake significantly. Down-regulation and recovery of D-oligo uptake induced by excess unlabeled D-oligo pretreatment occurred, but this was only partial. Studies with a digitonin equilibrium method and confocal microscopy indicated that intracellular localization of D-oligo was mainly in a vesicular compartment. More stable 3'-end-32P-labeled oligonucleotides were also used, to compare their uptake characteristics with those of the 5'-end-labeled oligonucleotides. The uptake characteristics for the two labels were similar in terms of saturability, greater D-oligo uptake, compared with S-oligo, and the inhibitory effect of p-chloromercuribenzoic acid; however, the stable 3'-end label showed greater uptake than the 5'-end label. Our findings suggest that D-oligo uptake by HCT-15 cells is mediated by endocytosis involving binding sites on the plasma membrane.


Non-parametric permutation thresholding for adaptive nonlinear beamformer analysis on MEG revealed oscillatory neuronal dynamics in human brain.
Adaptive nonlinear beamformer technique for analyzing magnetoencephalography (MEG) data has been proved to be powerful tool for both brain research and clinical applications. A general method of analyzing multiple subject data with a formal statistical treatment for the group data has been developed and applied for various types of MEG data. Our latest application of this method was frontal midline theta rhythm (Fmθ), which indicates focused attention and appears widely distributed over medial prefrontal areas in EEG recordings. To localize cortical generators of the magnetic counterpart of Fmθ precisely and identify cortical sources and underlying neural activity associated with mental calculation processing (i.e., arithmetic subtraction), we applied adaptive nonlinear beamformer and permutation analysis on MEG data. As a result, it was indicated that Fmθ is generated in the dorsal anterior cingulate and adjacent medial prefrontal cortex. Gamma event-related synchronization is as an index of activation in right parietal regions subserving mental subtraction associated with basic numerical processing and number-based spatial attention. Gamma desynchronization appeared in the right lateral prefrontal cortex, likely representing a mechanism to interrupt neural activity that can interfere with the ongoing cognitive task. We suggest that the combination of adaptive nonlinear beamformer and permutation analysis on MEG data is quite powerful tool to reveal the oscillatory neuronal dynamics in human brain.


Sleeve Lobectomy for Non-Small Cell Lung Cancers: Predictive CT Features for Resectability and Outcome Analysis.
OBJECTIVE. The purpose of this study was to investigate the CT features predictive of resectability in sleeve lobectomy (SL) for patients with non-small cell lung cancers (NSCLCs) and the outcomes of this procedure. MATERIALS AND METHODS. The study included 45 patients with NSCLCs who planned to undergo SL as a first-line surgical option and underwent SL or pneumonectomy between February 2014 and July 2017. The patients were dichotomized according to whether SL was achieved without residual disease. ROC curve and logistic regression analyses were performed to identify CT features predictive of resectability in SL without residual disease. For outcome analysis, postoperative complications and survival were compared between the two patient groups. RESULTS. SL was successfully performed in 25 patients (55.6%), whereas eight patients underwent SL but had a positive resection margin and 12 required pneumonectomy. The AUC value of the tumor-involved length of the salvage bronchus for SL without residual disease was 0.802, with 5 mm considered the optimal cutoff for this tumor-involved length. A tumor-involved length of 5 mm or less for the salvage bronchus representing the distal anastomosis end in SL was the sole significant predictor for resectability in SL without residual disease in logistic regression analysis (odds ratio, 16.0; 95% CI, 3.7-69.6; p < 0.001). The group of patients who underwent SL without residual disease tended to have a more favorable survival outcome than patients who underwent SL but had residual disease or those who underwent pneumonectomy (p = 0.110), but postoperative complication rates were similar between the two patient groups (20.0% vs 25.0%, respectively). CONCLUSION. A tumor-involved salvage bronchus length of 5 mm or less was a CT feature predictive of resectability in SL without residual disease, which may improve the survival of patients with NSCLCs.


The Modified Stroop Task Is Susceptible to Feigning: Stroop Performance and Symptom Over-endorsement in Feigned Test Anxiety.
Some researchers argue that the modified Stroop task (MST) can be employed to rule out feigning. According to these authors, modified Stroop interference effects are beyond conscious control and therefore indicative of genuine psychopathology. We examined this assumption using a within-subject design. In the first session, students (N = 22) responded honestly, while in the second session they were asked to read a vignette about test anxiety and then fake this condition. During both sessions, we administered an MST consisting of neutral, anxiety-related, and test anxiety-related words. Participants also completed the Self-Report Symptom Inventory (SRSI; Merten et al., 2016) that focuses on over-reporting of pseudosymptoms. Our feigning instructions were successful in that students succeeded in generating the typical MST effect by providing longer response latencies on anxiety related (r = 0.43) and test anxiety-related (r = 0.31) words, compared with neutral words. Furthermore, students endorsed significantly more pseudosymptoms on the SRSI (r = 0.62) in the feigning session than in the honest control condition. We conclude that the MST effect is not immune to feigning tendencies, while the SRSI provides promising results that require future research.