Effect of executive functioning, decision-making and self-reported impulsivity on the treatment outcome of pathologic gambling.
Impairments in self-regulatory behaviour reflect a deficit in executive functioning and decision-making, as well as higher levels of self-reported impulsivity, and may be involved in the development and maintenance of addictive disorders. We sought to explore the association between self-reported impulsivity and neurocognitive measures, and their association with treatment outcome in pathologic gambling. We assessed patients with pathologic gambling using executive functioning and decision-making tests and self-report measures of impulsivity. Patients underwent cognitive-behavioural therapy (CBT) for pathologic gambling. We included 88 patients (8% women) in our study. High self-reported extravagance was associated with poor performance in the Iowa Gambling Task (IGT)-ABCD version. High impulsiveness, low disorderliness, high exploratory excitability (trend), poor backward block span and poor IGT-EFGH scores (trend) predicted dropout. We observed no self-reported or neurocognitive predictors of relapse or number of treatment sessions attended. Most participants were slot-machine gamblers seeking treatment. No follow-up data and no control group were included in the study. The missing sample (i.e., individuals who were recruited and assessed in the pretreatment stage but who chose not to begin treatment) had higher extravagance scores than the final sample. Neurocognitive reward sensitivity was related to self-reported overspending behaviour. Self-regulatory impairments (especially rash impulsiveness and punishment sensitivity) and executive dysfunction predicted only dropout of CBT in participants with pathologic gambling. Different neurocognitive processes and personality traits might mediate treatment response to psychological therapy of pathologic gambling according to the specific target variable assessed.


Center-of-pressure regularity as a marker for attentional investment in postural control: a comparison between sitting and standing postures.
Postural control is a highly automatized basic activity that requires limited attentional investments. These investments have been shown to increase from balancing experts to controls, and from controls to persons with impaired postural control. Such between-subject comparisons led to a proposed direct relation between the regularity of center-of-pressure (COP) fluctuations and the amount of attention invested in posture. This study aims to expand this relation to a within-subject comparison of conditions that differ in balance demands. Specifically, more regular COP fluctuations were expected for standing than sitting, as stimulus-response reaction-time studies showed that the required attentional demands are lower for sitting than standing. COP registrations were made for fifteen healthy young adults in seated and standing postures. COP regularity was quantified with sample entropy. As expected, COP fluctuations were found to be more regular for standing than sitting, as evidenced by significantly lower sample entropy values. These findings expand the relation between COP regularity and the amount of attention invested in posture to postural tasks that vary in balance demands. An assessment of COP regularity may thus not only be instrumental in the examination of attentional investment in posture in between-subject designs, but also for different postures in within-subjects designs.


Lymphocyte chemoattractant factor induces CD4-dependent intracytoplasmic signaling in lymphocytes.
At present, a naturally occurring soluble ligand for CD4 has not been well described. There is much evidence to indicate that MHC class II molecules can bind to CD4; however, binding of intact class II molecules under physiologic conditions has been difficult to demonstrate. We investigated whether a previously reported human lymphokine, lymphocyte chemoattractant factor (LCF), which was described for its effects on human CD4+ lymphocytes and monocytes, could associate directly with CD4 and induce the generation of second messengers. In the present study, we demonstrate that CD4 affinity-purified natural and recombinant LCF induced a rise in intracellular calcium and increased inositol trisphosphate generation in normal human CD4+ lymphocytes and murine T cell hybridomas infected to express human CD4. Cell lines lacking human CD4 or expressing human CD4 molecules that lack the intracytoplasmic domain did not demonstrate a change in either calcium or inositol trisphosphate. The effect of LCF was blocked by coincubation with either anti-CD4 antibody or Fab fragments from anti-CD4 antibody. These studies demonstrate direct interaction of a lymphokine with CD4 and generation of second messengers as a result of the interaction.


Online Adaptive Magnetic Resonance-guided Radiation Therapy for Gynaecological Cancers: Preliminary Results of Feasibility and Outcome.
To present the preliminary results on the clinical utilisation of an online daily adaptive magnetic resonance-guided radiation therapy (MRgRT) for various gynaecological cancers. Twelve patients treated between September 2018 and June 2022 were included. Six patients (50%) were treated with pelvic radiation therapy followed by MRgRT boost as brachytherapy boost was ineligible or unavailable, three patients (25%) were treated with pelvic MRgRT followed by high dose rate brachytherapy, two patients (16.7%) were treated with only MRgRT, one patient (8.3%) was treated with linear accelerator-based radiation therapy followed by MRgRT boost for bulky iliac lymph nodes. The median age was 56.5 years (range 31-86 years). Eight patients (66.7%) had a complete response, three patients (25%) had a partial response and one patient (8.3%) died due to acute renal failure. The mean follow-up time was 11.2 months (range 3.1-42.6 months). The estimated 1-year overall survival was 88.9%. The median treatment time was 47 days (range 10-87 days). During external beam radiation therapy, 10 (83.3%) patients had concomitant chemoradiotherapy. Pelvic external beam radiation therapy doses for all cohorts were 45-50.4 Gy with a fraction dose of 1.8 Gy. The median magnetic resonance-guided boost dose was 32 Gy (range 20-50 Gy) and fraction doses ranged between 4 and 10 Gy. Three patients were treated with intracavitary high dose rate brachytherapy (26-28 Gy in four to five fractions). None of the patients had grade >3 late genitourinary toxicities. MRgRT is reliable and clinically feasible for treating patients with gynaecological cancers alone or in combination with brachytherapy with an acceptable toxicity and outcome. MRgRT boost could be an option when brachytherapy is not available or ineligible.


Idebenone inhibits catecholamine secretion through its blocking action on Ca2+ channels in cultured adrenal chromaffin cells.
The effect of idebenone, an agent improving cerebral metabolism, on catecholamine secretion was examined using primary cultures of bovine adrenal chromaffin cells. Catecholamine secretion evoked by acetylcholine was markedly inhibited by idebenone, and this effect was concentration-dependent. In contrast, other cerebral metabolism-improving agents, such as hopantenate and propentofylline, failed to cause any significant effect on the secretion in the same concentration range. Furthermore, idebenone inhibited the secretion evoked by high K+, veratridine, and Ba2+, but failed to inhibit the secretion evoked by the Ca(2+)-ionophore A23187. Idebenone also inhibited the radioactive Ca2+ uptake stimulated by acetylcholine or high K+ under the conditions in which its inhibitory action on the secretion was observed. Nifedipine, a typical voltage-dependent Ca2+ channel blocker, inhibited the secretion evoked by high K+, and this inhibitory action on the secretion was markedly reduced by the presence of idebenone. The present results suggest that idebenone may inhibit the influx of extracellular Ca2+ into the cells presumably through its blocking action on the voltage-dependent Ca2+ channels, resulting in the inhibition of catecholamine secretion in the adrenal chromaffin cell.


GAT1 (GABA:Na+:Cl-) cotransport function. Database reconstruction with an alternating access model.
We have developed an alternating access transport model that accounts well for GAT1 (GABA:Na+:Cl-) cotransport function in Xenopus oocyte membranes. To do so, many alternative models were fitted to a database on GAT1 function, and discrepancies were analyzed. The model assumes that GAT1 exists predominantly in two states, Ein and E(out). In the Ein state, one chloride and two sodium ions can bind sequentially from the cytoplasmic side. In the Eout state, one sodium ion is occluded within the transporter, and one chloride, one sodium, and one gamma-aminobutyric acid (GABA) molecule can bind from the extracellular side. When Ein sites are empty, a transition to the Eout state opens binding sites to the outside and occludes one extracellular sodium ion. This conformational change is the major electrogenic GAT1 reaction, and it rate-limits forward transport (i.e., GABA uptake) at 0 mV. From the Eout state, one GABA can be translocated with one sodium ion to the cytoplasmic side, thereby forming the *Ein state. Thereafter, an extracellular chloride ion can be translocated and the occluded sodium ion released to the cytoplasm, which returns the transporter to the Ein state. GABA-GABA exchange can occur in the absence of extracellular chloride, but a chloride ion must be transported to complete a forward transport cycle. In the reverse transport cycle, one cytoplasmic chloride ion binds first to the Ein state, followed by two sodium ions. One chloride ion and one sodium ion are occluded together, and thereafter the second sodium ion and GABA are occluded and translocated. The weak voltage dependence of these reactions determines the slopes of outward current-voltage relations. Experimental results that are simulated accurately include (a) all current-voltage relations, (b) all substrate dependencies described to date, (c) cis-cis and cis-trans substrate interactions, (d) charge movements in the absence of transport current, (e) dependencies of charge movement kinetics on substrate concentrations, (f) pre-steady state current transients in the presence of substrates, (g) substrate-induced capacitance changes, (h) GABA-GABA exchange, and (i) the existence of inward transport current and GABA-GABA exchange in the nominal absence of extracellular chloride.


Optimal Antithrombotic Regimens for Patients With Atrial Fibrillation Undergoing Percutaneous Coronary Intervention: An Updated Network Meta-analysis.
Antithrombotic treatment in patients with atrial fibrillation (AF) and percutaneous coronary intervention (PCI) presents a balancing act with regard to bleeding and ischemic risks. To evaluate the safety and efficacy of 4 antithrombotic regimens by conducting an up-to-date network meta-analysis and to identify the optimal treatment for patients with AF undergoing PCI. Online computerized database (MEDLINE). Five randomized studies were included (N = 11 542; WOEST, PIONEER AF-PCI, RE-DUAL PCI, AUGUSTUS, ENTRUST-AF PCI). The Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guidelines were used in this network meta-analysis, in which bayesian random-effects models were applied. The data were analyzed from September 9 to 29, 2019. The primary safety outcome was thrombolysis in myocardial infarction (TIMI) major bleeding and the primary efficacy outcome was trial-defined major adverse cardiovascular events (MACE). The total number of participants included in the study was 11 532. The mean age of the participants ranged from 70 to 72 years, 69% to 83% were male, 20% to 26% were female, and the participants were predominantly white (>90%). Compared with vitamin K antagonists (VKA) plus dual antiplatelet therapy (DAPT) (reference), the odds ratios (ORs) (95% credible intervals) for TIMI major bleeding were 0.57 (0.31-1.00) for VKA plus P2Y12 inhibitor, 0.69 (0.40-1.16) for non-VKA oral anticoagulant (NOAC) plus DAPT, and 0.52 (0.35-0.79) for NOAC plus P2Y12 inhibitor. For MACE, using VKA plus DAPT as reference, the ORs (95% credible intervals) were 0.97 (0.64-1.42) for VKA plus P2Y12 inhibitor, 0.95 (0.64-1.39) for NOAC plus DAPT, and 1.03 (0.77-1.38) for NOAC plus P2Y12 inhibitor. The findings of this study suggest that an antithrombotic regimen of VKA plus DAPT should generally be avoided, because regimens in which aspirin is discontinued may lead to lower bleeding risk and no difference in antithrombotic effectiveness. The use of a NOAC plus a P2Y12 inhibitor without aspirin may be the most favorable treatment option and the preferred antithrombotic regimen for most patients with AF undergoing PCI.


Interaction of temperature, humidity, driver preferences, and refrigerant type on air conditioning compressor usage.
Recent studies have shown large increases in vehicle emissions when the air conditioner (AC) compressor is engaged. Factors that affect the compressor-on percentage can have a significant impact on vehicle emissions and can also lead to prediction errors in current emissions models if not accounted for properly. During 1996 and 1997, the University of California, Riverside, College of Engineering-Center for Environmental Research and Technology (CE-CERT) conducted a vehicle activity study for the California Air Resources Board (CARB) in the Sacramento, CA, region. The vehicles were randomly selected from all registered vehicles in the region. As part of this study, ten vehicles were instrumented to collect AC compressor on/off data on a second-by-second basis in the summer of 1997. Temperature and humidity data were obtained and averaged on an hourly basis. The ten drivers were asked to complete a short survey about AC operational preferences. This paper examines the effects of temperature, humidity, refrigerant type, and driver preferences on air conditioning compressor activity. Overall, AC was in use in 69.1% of the trips monitored. The compressor was on an average of 64% of the time during the trips. The personal preference settings had a significant effect on the AC compressor-on percentage but did not interact with temperature. The refrigerant types, however, exhibited a differential response across temperature, which may necessitate separate modeling of the R12 refrigerant-equipped vehicles from the R134A-equipped vehicles. It should be noted that some older vehicles do get retrofitted with new compressors that use R134A; however, none of the vehicles in this study had been retrofitted.


Dietary calcium and magnesium intake and mortality: a prospective study of men.
The authors examined the association of dietary calcium and magnesium intake with all-cause, cardiovascular disease (CVD), and cancer mortality among 23,366 Swedish men, aged 45-79 years, who did not use dietary supplements. Cox proportional hazards regression models were used to estimate the multivariate hazard ratios and 95% confidence intervals of mortality. From baseline 1998 through December 2007, 2,358 deaths from all causes were recorded in the Swedish population registry; through December 2006, 819 CVD and 738 cancer deaths were recorded in the Swedish cause-of-death registry. Dietary calcium was associated with a statistically significant lower rate of all-cause mortality (hazard ratio (HR) = 0.75, 95% confidence interval (CI): 0.63, 0.88; P(trend) < 0.001) and a nonsignificantly lower rate of CVD (HR = 0.77, 95% CI: 0.58, 1.01; P(trend) = 0.064) but not cancer mortality (HR = 0.87, 95% CI: 0.65, 1.17; P(trend) = 0.362) when the highest intake tertile (mean = 1,953 mg/day; standard deviation (SD), 334) was compared with the lowest (990 mg/day; SD, 187). Dietary magnesium intake (means of tertiles ranged from 387 mg/day (SD, 31) to 523 mg/day (SD, 38) was not associated with all-cause, CVD, or cancer mortality. This population-based, prospective study of men with relatively high intakes of dietary calcium and magnesium showed that intake of calcium above that recommended daily may reduce all-cause mortality.


Iron-containing lipoprotein SiaA in SiaABC, the primary heme transporter of Streptococcus pyogenes.
The cell-surface lipoprotein SiaA, a component of the SiaABC transporter, acts as the primary receptor for heme in the infamous human pathogen Streptococcus pyogenes. However, little is known about the molecular mechanism of heme binding and release as well as the role of heme-binding ligands that contribute to the uptake of heme into the pathogenic bacteria. The present report aims to clarify the coordination properties of heme iron in SiaA. By substitution of either Met79 or His229 with alanine, the mutant M79A and H229A proteins display significantly decreased heme-binding affinity and substantially increased heme-release rates, as compared with wild-type SiaA protein. Both fluorescence and circular dichroism spectra indicated that heme binding results in alterations in the secondary structure of the protein. Heme release from SiaA is a stepwise process in which heme dissociates firstly from Met79 and then from His229 with distinct conformational changes. His229 may serve as an anchor for heme binding in SiaA and thus may play a major role in the stability of the coordination between heme and the protein.


The feasibility of light microscopic measurements of intercellular spaces in squamous epithelium in the lower-esophagus of GERD patients.
The study aims to determine whether light microscopy can be used to accurately measure the diameters of intercellular spaces between squamous epithelial cells in the lower esophagus, and whether changes in this outcome measure can be used as a diagnostic marker for gastroesophageal reflux disease (GERD). The study has two parts. Part 1 involves 42 asymptomatic controls and 119 patients with typical symptoms of GERD, including 58 with erosive esophagitis (EE), and 61 patients with nonerosive gastroesophageal reflux disease (NERD). All biopsies were taken from the lower esophagus. All samples were observed using an immersion objective, after which diameters were measured by computer-assisted morphometry. Part 2 involves 61 individuals who were randomly selected from part 1, including 19 controls, 13 with NERD and 29 with EE. Diameter measurements using both light microscopy and transmission electron microscopy (TEM) were performed for samples of 61 individuals. Samples from a total of 61 individuals (31 male, 30 female, mean age 44.3 ± 16.0 years) were observed using both light microscopy and TEM. Both methods showed significant differences between control and disease groups; the outcomes from the two methods had a certain correlation (r = 0.605, P = 0.000). Morphometric analysis of all 161 individuals (83 males, 78 females, mean age 41.4 ± 15.7) showed mean diameters from light microscopy to be 0.58 ± 0.16 µm for controls, 1.07 ± 0.30 µm for NERD, and 1.29 ± 0.20 µm for EE; differences between control and disease groups were significant (P<0.05). The optimal cut-off value from receiver operator characteristic analysis was 0.85 µm. Diagnoses were validated using the combination of symptoms of GERD, endoscopy, and 24 h ambulatory pH monitoring as the gold standard. At the optimal cutoff, sensitivity was 93.3% and specificity was 100%. The diameters of the intercellular spaces in squamous epithelium of lower esophagus from controls and in patients with GERD can be quantitatively measured using light microscopy. Dilated diameters can serve as a sensitive, specific, and objective indicator for diagnosis of GERD.


Suppression of cell-mediated immune responses to listeria infection by repeated exposure to diesel exhaust particles in brown Norway rats.
Diesel exhaust particles (DEP) have been shown to alter pulmonary immune responses to bacterial infection. Exposure of rats to 100 mg/m(3) DEP for 4 h was found to aggravate Listeria monocytogenes(Listeria) infection at 3 days postinfection, but the bacteria were largely cleared at 7 days postinfection due to the development of a strong T cell-mediated immunity. In the present study, we examined the effects of repeated DEP exposure at lower doses on pulmonary responses to bacterial infection. Brown Norway rats were exposed to DEP by inhalation at 20.62 +/- 1.31 mg/m 3 for 4 h/day for 5 days, followed by intratracheal inoculation with 100,000 Listeria at 2 h after the last DEP exposure. DEP-exposed rats showed a significant increase in lung bacterial load at both 3 and 7 days postinfection. The repeated DEP exposure was shown to suppress both the innate, orchestrated by alveolar macrophages (AM), and T cell-mediated responses to Listeria. DEP inhibited AM production of interleukin- (IL-) 1beta, tumor necrosis factor- (TNF-) alpha, and IL-12 but enhanced Listeria-induced AM production of IL-10, which has been shown to prolong the survival of intracellular pathogens such as Listeria. DEP exposure also suppressed the development of bacteria-specific lymphocytes from lung-draining lymph nodes, as indicated by the decreased numbers of T lymphocytes and their CD4(+) and CD8(+) subsets. Furthermore, the DEP exposure markedly inhibited the Listeria-induced lymphocyte secretion of IL-2 at day 7, IL-10 at days 3 and 7, and interferon- (IFN-) gamma at days 3 to 10 postinfection when compared to air-exposed controls. These results show a sustained pattern of downregulation of T cell-mediated immune responses by repeated low-dose DEP exposure, which is different from the results of a single high-dose exposure where the acute effect of DEP aggravated bacteria infection but triggered a strong T cell-mediated immunity.


Effects of human recombinant interleukin-1beta on canine articular chondrocytes in three-dimensional culture.
To determine the effects of interleukin (IL)-1beta on matrix synthesis and degradation by chondrocytes cultured in a 3-dimensional (3-D) gel medium. Chondrocytes from 7 dogs. Articular chondrocytes were harvested and cultured in 3-D gel medium alone or with 10 or 20 ng IL-1beta/ml that was added beginning on day 0, 3, 6, or 9. On days 3, 6, 12, and 20 of 3-D culture, samples of the liquid medium were evaluated for glycosaminoglycan (GAG), prostaglandin E2 (PGE2), and matrix metalloprotease (MMP)-3 content. The 3-D plug in each well was evaluated for histologic characteristics of viability, cell morphology, and proteoglycan staining, immunohistochemically stained for collagen type II, and spectrophotometrically analyzed for GAG content. Significant differences for all variables were detected between controls and each IL-1beta group, among groups with different IL-1beta concentrations, and among groups with IL-1beta added at various time points. Chondrocytes exposed to IL-1beta had loss of GAG, increased PGE2 and MMP-3 concentrations, and lack of collagen type-II synthesis. These IL-1beta effects appeared to be time and concentration dependent. Addition of IL-1beta to chondrocytes in 3-D gel medium results in time- and concentration-dependent effects on matrix synthesis and degradation and provides an appropriate in vitro model for many of the pathophysiologic events associated with osteoarthritis.


Greater role for Atlantic inflows on sea-ice loss in the Eurasian Basin of the Arctic Ocean.
Arctic sea-ice loss is a leading indicator of climate change and can be attributed, in large part, to atmospheric forcing. Here, we show that recent ice reductions, weakening of the halocline, and shoaling of the intermediate-depth Atlantic Water layer in the eastern Eurasian Basin have increased winter ventilation in the ocean interior, making this region structurally similar to that of the western Eurasian Basin. The associated enhanced release of oceanic heat has reduced winter sea-ice formation at a rate now comparable to losses from atmospheric thermodynamic forcing, thus explaining the recent reduction in sea-ice cover in the eastern Eurasian Basin. This encroaching "atlantification" of the Eurasian Basin represents an essential step toward a new Arctic climate state, with a substantially greater role for Atlantic inflows.


Global mental health: perspectives from Ethiopia.
Global mental health (GMH) advocates for access to and the equitable provision of mental health care. Although the treatment gap is a useful construct to measure access and equitability of care, it fails to communicate the real-life consequences of the treatment gap and the urgent need to address care disparities. The aim of this article is to present a perspective on the practical application of the principles of GMH to understand the real-life impact of the treatment gap and the approaches taken to improve treatment coverage in Ethiopia. A case study method is used. Multiple international collaborations undertaken in Ethiopia and facilitated by GMH to improve care, capacity, and the evidence base for increased treatment coverage are described briefly. A series of steps taken at the local and national levels to address the treatment gap are highlighted. The stories of two patients are also presented to illustrate the real-life consequences of the treatment gap and the potential transformational impact of addressing the treatment gap on patients, families, and communities. GMH has a key role to play in addressing the treatment gap, which improves the life of people with mental disorders, their families, and their communities. However, national-level policy support and coordination are essential for any realistic improvement in treatment coverage. The reflections offered through the case examples may have utility in similar low-income settings.


Analysis of human and swine hepatitis E virus (HEV) isolates of genotype 3 in Japan that are only 81-83 % similar to reported HEV isolates of the same genotype over the entire genome.
Full-length sequences were determined for a human hepatitis E virus (HEV) isolate (HE-JA04-1911) and two swine HEV isolates (swJ8-5 and swJ12-4) that belong to one of three clusters within genotype 3 in Japan and are close to Spanish isolates according to their partial sequences. The three HEV isolates were 89.7-92.9 % identical to each other, but only 80.7-83.0 % similar to 21 HEV strains of the same genotype isolated in Canada, Kyrgyzstan, the USA and Japan over their entire genome. On comparison with HEV isolates whose partial sequence is known, the HE-JA04-1911, swJ8-5 and swJ12-4 isolates segregated into a phylogenetic cluster consisting of human and swine HEV isolates in Japan and the UK, with identities of 89.8-100 % and 87.9-92.4 %, respectively. Genotype 3 HEV isolates were found to be markedly heterogeneous. The UK-isolate-like HEV strains in Japan may have originated from the UK via the importation of pigs since 1900.


Intolerance of uncertainty and conditioned place preference in opioid addiction.
Several personality factors have been implicated in vulnerability to addiction by impacting learning and decision making. One such factor is intolerance of uncertainty (IU), the tendency to perceive uncertain situations negatively and avoid them. Conditioned place preference (CPP), which compares preference for contexts paired with reward, has been used to examine the motivation for both drug and non-drug rewards. However, preference for locations associated with non-drug reward, as well as the potential influence of IU, has not been thoroughly studied in individuals with addiction. In the current study, we examined CPP using a computer-based task in a sample of addicted individuals undergoing opioid maintenance treatment and never-addicted controls. Patients were confirmed to have higher IU than controls. In the CPP task, the two groups did not differ in overall time spent in the previously-rewarded context. However, controls were more likely than patients to immediately return to this context. Contrary to our predictions, IU was not a significant predictor of preference for the previously-rewarded context, although higher IU in controls was associated with a higher number of rewards obtained in the task. No such relationship was found in patients.


A Novel Approach to Increase Glial Cell Populations in Brain Microphysiological Systems.
Brain microphysiological systems (bMPS) recapitulate human brain cellular architecture and functionality more closely than traditional monolayer cultures and have become increasingly relevant for the study of neurological function in health and disease. Existing 3D brain models vary in reflecting the relative populations of different cell types present in the human brain. Most models consist mainly of neurons, while glial cells represent a smaller portion of the cell populations. Here, by means of a chemically defined glial-enriched medium (GEM), an improved method to expand the population of astrocytes and oligodendrocytes without compromising neuronal differentiation in bMPS, is presented. An important finding is that astrocytes also change in morphology when cultured in GEM, more closely recapitulating primary culture astrocytes. GEM bMPS are electro-chemically active and show different patterns of calcium staining and flux. Synaptic vesicles and terminals observed by electron microscopy are also present. No significant changes in neuronal differentiation are observed by gene expression, however, GEM enhanced neurite outgrowth and cell migration, and differentially modulated neuronal maturation in two different cell lines. These results have the potential to significantly improve functionality of bMPS for the study of neurological diseases and drug discovery, contributing to the unmet need for safe human models.


In Vitro Evaluation of Cancer Cell Immunogenicity and Antigen-Specific T-Cell Cytotoxicity by Flow Cytometry.
A cardinal principle of oncoimmunology is that cancer cells can be eliminated by tumor-infiltrating cytotoxic CD8 T lymphocytes. This has been widely demonstrated during the last 20 years and also recently harnessed for therapy. However, emerging evidence indicates that even neoplasms showing striking initial responses to conventional and targeted (immuno)therapies often acquire resistance, resulting in tumor relapse, increased aggressiveness, and metastatization. Indeed, tumors are complex ecosystems whose malignant and nonmalignant cells, constituting the tumor microenvironment, constantly interact and evolve in space and time. Together with patient's own genetic factors, such environmental interplays may curtail antitumor immune responses leading to cancer immune evasion and natural/acquired (immuno)therapy resistance. In this context, cancer stem cells (CSCs) are thought to be the roots of therapy failure. Flow cytometry is a powerful technology that finds extensive applications in cancer biology. It offers several unique advantages as it allows the rapid, quantitative, and multiparametric analysis of cell populations or functions at the single-cell level. In this chapter, we discuss a two-color flow cytometric protocol to evaluate cancer cell immunogenicity by analyzing the proliferative and tumor-killing potential of ovalbumin (OVA)-specific CD8 OT-1 T cells exposed to OVA-expressing MCA205 sarcoma cells and their CSC counterparts.


Alpha 2-adrenoreceptor stimulation does not inhibit L-type calcium channels in mouse pancreatic beta-cells.
The effects of alpha 2-adrenergic stimulation on the Ca(2+)-current in mouse pancreatic beta-cells were investigated using the patch-clamp technique. When using the conventional whole-cell recording configuration (dialysis of cell interior with pipette solution), addition of adrenaline (1 microM) or the alpha 2-adrenergic agonist clonidine (5 microM) failed to reduce the Ca(2+)-current, irrespective of whether intracellular GTP (or GTP gamma S) was present or not and at both physiological (1.3 mM) and elevated (10.2 mM) Ca(2+)-concentrations. In fact, in the absence of added guanine nucleotides, the agonists tended to increase the Ca(2+)-current amplitude in the presence of the higher Ca(2+)-concentration. Ca(2+)-channel activation measured at 1.3 mM Ca2+ was not affected by clonidine. Half-maximal activation was observed at approximately -20 mV. In addition, when Ca(2+)-currents were recorded from intact beta-cells, using the perforated patch whole-cell configuration, clonidine (1 microM) also failed to detectably affect the Ca(2+)-current. It is therefore suggested that the inhibition of beta-cell electrical activity and insulin-secretion produced by alpha 2-adrenoreceptor stimulation does not result from suppression of the L-type Ca(2+)-current.


IMMUNOLOGIC RELATION OF STREPTOCOCCAL AND TISSUE ANTIGENS. III. PRESENCE IN HUMAN SERA OF STREPTOCOCCAL ANTIBODY CROSS-REACTIVE WITH HEART TISSUE. ASSOCIATION WITH STREPTOCOCCAL INFECTION, RHEUMATIC FEVER, AND GLOMERULONEPHRITIS.
Sera from patients with recent streptococcal infection or non-suppurative sequelae exhibit with variable frequency a precipitin reaction in agar gel with a partially purified streptococcal antigen which has been shown previously to be immunologically related to human heart tissue. This precipitin could be absorbed from sera with normal human heart tissue homogenates but not with homogenates of other organs. Demonstration of this cross-reaction by heart absorption was found dependent both upon the serologic properties of individual sera and the nature or state of purification of the streptococcal product employed as test antigen. Evidence was obtained of a close association of heart-related and non-heart-related antigenic determinants in partially purified preparations of the streptococcal antigen by both gel diffusion and immunoelectrophoresis. On immunoelectrophoretic analysis, cross-reactive antigen exhibited a more rapid mobility toward the anode than M protein. It was destroyed by digestion with trypsin, pepsin, and chymotrypsin. Based on specific absorption tests with a Type 5 and Type 19 strain, the antigen was localized to cell walls and to a lesser extent to cell membranes of these strains. Precipitating activity related to cross-reactive antibody was localized to the immunoglobulin zone in immunoelectrophoresis. Reactive sera showed diminution or loss of serological activity following heat inactivation at 56 degrees C or after prolonged storage at 4 degrees C. Sera containing cross-reactive precipitating antibody exhibited an immunofluorescent reaction with sarcolemma of cardiac myofibers, which was inhibited by streptococcal cross-reactive antigen. By this inhibition test, the immunofluorescent reaction related to cross-reactive antibody could be distinguished from that due to other heart-reactive factors. Antibody to streptococcal cross-reactive antigen defined by precipitation-absorption tests was observed in 24 per cent of patients with recent history of uncomplicated streptococcal infection and in the majority of patients with acute rheumatic fever, rheumatic heart disease, or acute glomerulonephritis. It was observed rarely in patients with non-streptococcal related disease. These data provide evidence that induction of cross-reactive autoantibody to heart in certain individuals is associated with streptococcal infection.


Comparison of injectable iron complexes in their ability to iron load tissues and to induce oxidative stress.
Iron and copper homeostasis have been studied in various tissues after iron-loading with the polynuclear ferric hydroxide carbohydrate complexes, iron dextran, iron polymaltose, iron sucrose and iron gluconate for four weeks. There were significant increases in the iron content of the different rat tissues compared to controls, with the exception of the brain, which showed no change in its iron content following iron loading. However, the level of iron loading in the different tissues varied according to the preparation administered and only iron dextran was able to significantly increase the iron content of both broncho-alveolar macrophages and heart. The hepatic copper content decreased with iron loading, although this did not reach significance. However the copper content did not alter in the iron loaded broncho-alveolar macrophages. Despite such increases in hepatic iron content, there was little evidence of changes in oxidative stress, the activities of cytosolic (apart from iron dextran) or mitochondrial hepatic superoxide dismutase, SOD, were similar to that of the control rats, confirming the fact that the low reduction potential of these compounds prevents the reduction of the ferric moiety. It was not necessary for macrophages to significantly increase their iron content to initiate changes in NO* release. Iron gluconate and iron sucrose increased NO* release, while iron polymaltose and iron dextran decreased NO* release although only the latter iron preparation significantly increased their iron content. It may be that the speciation of iron within the macrophage is an important determinant in changes in NO* release after ex vivo stimulation. We conclude that tissues loaded with iron by such polynuclear iron complexes have variable loading despite the comparable iron dose. However, there was little evidence for participation of the accumulated iron in free radical reactions although there was some evidence for alteration in immune function of broncho-alveolar macrophages.


Organic-inorganic hybrid 1-D double chain heteropolymolybdates constructed from plenary Keggin germanomolybdate anions and hepta-nuclear Cu-RE-pic heterometallic clusters.
Two unprecedented organic-inorganic hybrid 1-D double chain germanomolybdates containing hepta-nuclear Cu-RE-pic heterometallic clusters [NH4]2[RE(H2O)5]2[Cu(pic)2]2[Cu(pic)2(H2O)2]3[α-GeMo12O40]2·22H2O [RE = La3+ (1), Ce3+ (2), and Hpic = 2-picolinic acid] were successfully obtained by the stepwise self-assembly strategy via the conventional solution method. The most striking structural feature of 1 and 2 is that the two plenary Keggin [α-GeMo12O40]4- polyoxoanions are joined by an organic-inorganic hybrid hepta-nuclear Cu-RE-pic {[RE(H2O)5]2[Cu(pic)2(H2O)2]3[Cu(pic)2]2}6+ heterometallic moiety. What is more interesting is that their adjacent structural units are connected together by {Cu(pic)2} bridges, forming a 1-D extended double chain architecture. Furthermore, the adsorption capacity of 1 toward dyes in aqueous solutions was deeply investigated. It is fascinating that 1 shows a good adsorption capacity toward basic violet 3 (BV 3) in aqueous solutions and the adsorption kinetics conforms to the second-order kinetic model.


What is the effect of additional physiotherapy on sitting balance following stroke compared to standard physiotherapy treatment: a systematic review.
Sitting balance dysfunction is commonly experienced following stroke. Physiotherapists utilize interventions to address this problem but it is unclear whether treatment type, target or practice intensity may affect outcomes. To compare the effects of standard physiotherapy to standard physiotherapy plus an additional physiotherapy treatment after stroke. The databases of Cochrane Library, CINAHL, Embase, Ovid Medline, AMED, and the Physiotherapy Evidence Database (PEDro) up to December 2014 were searched. Randomized controlled trials in English reported in peer-reviewed journals regarding the effect of additional physiotherapy on sitting balance were retrieved. The PEDro scale was used to assess study quality. Eleven studies met inclusion criteria. Nine targeted the ICF (International Classification of Function, Disability and Health) domain of Activity. The Trunk control test (TCT) was used as a primary outcome measure in five studies, and the Trunk Impairment Scale (TIS) was used in four. There was a significant effect (mean difference = 1.67, 95% CI = 0.54-2.80) favoring intervention, as measured by the TIS. There was no evidence to support the effect of additional treatment on sitting balance as measured by the TCT (mean difference = - 1.53, 95% CI = - 9.37 to 6.32). The current evidence supports strategies that target deficits at the activity level and increase total treatment time. The TIS is most responsive as a measure of treatment efficacy. Further research is required using recommended outcome measures to facilitate generation of a minimum data set and data pooling.


Does the gender of the bully/victim dyad and the type of bullying influence children's responses to a bullying incident?
Children's responses to bullying are context related; they will vary depending on the specific bullying episode. The aim of the present study was to explore whether children's responses to bullying vary depending on the gender of the bully and victim and the type of bullying portrayed. In total, 437 children aged 9-11 years from four primary schools in the UK took part in the study. Each child read a story about one child bullying another. There were 12 different versions of the story, varying the type of bullying (verbal, physical, or relational/indirect) and the gender of the bully and victim (i.e., male bully-female victim, female bully-male victim, male bully-male victim, female bully-female victim). Each child was randomly allocated to one of the 12 stories. After reading the story the children then responded to a series of questions to assess their perceptions of the victim and bully and situation. Overall females liked the bully more than males; females also reported liking the female victim more than the male victim and females were more likely than males to intervene with a female victim. The bullying was viewed as more serious, more sympathy was shown to the victim, and there was a greater likelihood of intervention when the incident involved a female bully. There was less liking for the bully if the situation involved a female victim of physical bullying. The findings are explained in terms of social identity theory and social norms about typical male and female behavior.


Aggression, impulsivity, and suicide behavior: a review of the literature.
This article reviews the literature on the association between impulsivity aggression and suicide. The key words impulsivity, aggression, and suicide were entered into the pubmed, psychlit, and proqest databases. Significant articles were scrutinized for relevant information. Impulsivity and aggression are highly correlated with suicidal behavior across psychiatric samples, nosological borders, and non-psychiatric populations. Impulsivity and aggression are related but the nature of this relationship remains unclear. The literature is confusing and contradictory. This is probably due to the difficulty in defining and separating out these concepts and the fact that there is much overlap between them. Future research should aim at clarifying and refining these concepts as well as their link to all the different forms of suicidal behavior.


Decisions Regarding Newborn Feeding by Latina Mothers.
This study examined the factors that influence Latina mothers' decisions to initiate breastfeeding, formula feeding, or both, and when a given neonatal feeding method is decided, in order to assess the optimum time for healthcare providers to discuss nutrition decisions. A Spanish-speaking health care provider orally administered a 24-question survey to self-identified Latina mothers in the Newborn Nursery at North Carolina Children's Hospital after delivery. A total of 152 Latina mothers, aged 17 to 43 years, were interviewed from September 2014 to September 2015. Three decisive factors resulted from multivariate analysis regarding a participant's decision to exclusively breastfeed: age (27.3 years or younger); deciding about feeding method before pregnancy; and believing that exclusively breastfeeding is best for an infant's growth and development. Overall, 95% of Latina mothers made their decision regarding their infant's nutrition before delivery. Newborn nurseries have trended toward the allocation of substantial resources to the mission of enhancing the likelihood of breastfeeding initiation and success in the newborn period. Perhaps a reallocation of some of these resources for educational interventions earlier in pregnancy should be considered.


Separate Polycomb Response Elements control chromatin state and activation of the vestigial gene.
Patterned expression of many developmental genes is specified by transcription factor gene expression, but is thought to be refined by chromatin-mediated repression. Regulatory DNA sequences called Polycomb Response Elements (PREs) are required to repress some developmental target genes, and are widespread in genomes, suggesting that they broadly affect developmental programs. While PREs in transgenes can nucleate trimethylation on lysine 27 of the histone H3 tail (H3K27me3), none have been demonstrated to be necessary at endogenous chromatin domains. This failure is thought to be due to the fact that most endogenous H3K27me3 domains contain many PREs, and individual PREs may be redundant. In contrast to these ideas, we show here that PREs near the wing selector gene vestigial have distinctive roles at their endogenous locus, even though both PREs are repressors in transgenes. First, a PRE near the promoter is required for vestigial activation and not for repression. Second, only the distal PRE contributes to H3K27me3, but even removal of both PREs does not eliminate H3K27me3 across the vestigial domain. Thus, endogenous chromatin domains appear to be intrinsically marked by H3K27me3, and PREs appear required to enhance this chromatin modification to high levels at inactive genes.


Co-administration study in South African infants of a live-attenuated oral human rotavirus vaccine (RIX4414) and poliovirus vaccines.
A double-blind, placebo-controlled phase II trial (e-Track 444563-014/NCT00346892) was conducted in South Africa to evaluate the co-administration of RIX4414 (live-attenuated human G1P[8] rotavirus vaccine) and oral poliovirus vaccine (OPV) administered simultaneously. Healthy infants (n=450) were randomized into three groups (RIX4414+OPV, RIX4414+IPV or Placebo+OPV) to receive two oral doses of RIX4414/placebo with OPV or IPV using two vaccination schedules (6-10 weeks and 10-14 weeks). Serum anti-rotavirus IgA antibodies (ELISA) and neutralizing antibodies (micro-neutralization assay) to poliovirus serotypes 1, 2 and 3 were measured. Co-administration of RIX4414 with OPV did not result in a decrease in the high sero-protection rates against poliovirus serotypes 1, 2 and 3 detected after the third OPV dose (98-100%). The anti-rotavirus IgA antibody sero-conversion rates were higher for the 10-14 weeks schedule (55-61%) compared to the 6-10 weeks schedule (36-43%). Solicited symptoms were reported at similar rates between RIX4414 and placebo groups and no serious adverse events related to RIX4414 were reported. This study provided evidence that RIX4414 can be co-administered with routine EPI immunizations including OPV and that two doses of RIX4414 were well tolerated and immunogenic in South African infants.


[The dynamics of cardiac contraction in dogs with alloxan diabetes].
Development of alloxan diabetes was accompanied in dogs by regular changes in the phasic structure of the cardiac contractions. Direction of the changes in the indices of phasic analysis in the groups of dogs with an average (up to 144.3 +/- 12.5 mg%) and marked (up to 258.0 +/- 22.8 mg%) increase in the blood sugar level was the same, but their expression increased with the elevation of hyperglycemia. In marked hyperglycemia there was established an increase (in comparison with normal values) of the index of myocardial tension, a shortening of the period of ejection, a decrease of the mechanical coefficient and of the intrasystolic index, this coinciding with the character of changes of the corresponding indices in the patients suffering from diabetes mellitus, depending on the severity of the disease. The data obtained pointed to the significant influence of metabolic changes (assessed by hyperglycemia level) on the contractile function of the heart.


[Early experiences with the total mini hip-joint endoprosthesis (St. George Mini model) in dysplasia coxarthrosis].
The results of 106 patients with osteoarthritis secondary to severe congenital hip dysplasia and dislocations of the hip operated at the Endo-Klinik were evaluated after an average follow up of 21 months. 142 "St. Georg-Mini" total hips were implanted employing Refobacin-Palacos R. The clinical results were graded as "combined performance" which considered hip pain, walking distance, hip mobility and walking aids. In 93% of the cases "combined performance" was improved at follow up and 91.7% of the patients were satisfied with the result. The early (during hospitalization) and late (after discharge from hospital) complication rate which had an influence on the result at follow up was 2.1% (n = 3) and 3.5% (n = 4) respectively. There were no deep infections. An extended indication for total hip arthroplasty also for younger patients with severe osteoarthritis secondary to hip dysplasia is discussed and can be advocated.


6-Thioguanine inhibition of parathyroid hormone-related protein expression is mediated by GLI2.
Breast cancer cells frequently metastasize to bone, where they up-regulate their expression of the transcription factor GLI2 and the downstream osteolytic factor parathyroid hormone-related protein (PTHrP). The guanosine nucleotide 6-thioguanine (6-TG) inhibits PTHrP expression and blocks osteolytic bone destruction in mice inoculated with bone metastatic cells; however, the mechanism by which 6-TG inhibits PTHrP remains unclear. We hypothesized that 6-TG inhibition of PTHrP is mediated through GLI2 signaling. Human MDA-MB-231 breast cancer cells and RWGT2 squamous-cell lung carcinoma cells were treated with 100 μM 6-TG and examined for GLI2 mRNA expression and stability by Q-PCR, promoter activity by luciferase assay, and protein expression by Western blot. 6-TG significantly blocked GLI2 mRNA and protein expression, but did not affect stability. Additionally, 6-TG directly inhibited GLI2 promoter activity, and when cells were transfected with constitutively expressed GLI2, the inhibitory effect of 6-TG on PTHrP expression was abolished. Taken together, these data indicate that 6-TG regulates PTHrP in part through GLI2 transcription, and therefore the clinical use of 6-TG or other guanosine nucleotides may be a viable therapeutic option in tumor types expressing elevated levels of GLI proteins.


Effects of acute exercise on markers of inflammation in pediatric chronic kidney disease: a pilot study.
Children and adolescents with chronic kidney disease (CKD) are chronically exposed to high levels of inflammation, placing them at an increased risk of secondary health complications. Regular exercise may represent an effective therapy to reduce inflammation. The aims of this pilot study were to determine the effects of acute exercise on inflammation and immune cell counts in CKD. Nine children and adolescents (4 males) with CKD stages III-V performed a graded exercise test to determine peak oxygen uptake (VO2peak). Following a 10-min break, participants cycled for 20 min at 50 % of VO2peak. Blood samples were collected before and after the exercise period for the determination of complete blood counts, natural killer cells (NK(bright), NK(dim)) and circulating progenitor cell (CPC) counts, as well as interleukin-6 (IL-6), and tumor necrosis factor-α (TNF-α) concentrations. Complete blood counts and NK(dim) cell and CPC counts were unchanged with exercise. Following exercise, NK(bright) cell counts increased (7.4 ± 4.3 vs. 12.2 ± 8.3 × 10(6) cells/L; p = 0.02), while trends were observed for an increase in IL-6 (2.1 ± 2.2 vs. 2.7 ± 2.6 pg/mL; p = 0.08), decrease in TNF-α (4.5 ± 1.2 vs. 4.2 ± 1.0 pg/mL; p = 0.08) and an increase in the IL-6:TNF-α ratio (0.6 ± 0.7 vs. 0.8 ± 0.8; p = 0.07). Our findings suggest that acute exercise may create an anti-inflammatory environment in children and adolescents with CKD stages III-V.


Highly Eribulin-resistant KBV20C Oral Cancer Cells Can Be Sensitized by Co-treatment with the Third-generation P-Glycoprotein Inhibitor, Elacridar, at a Low Dose.
Eribulin mesylate, also called Halaven® (HAL), was recently developed as a microtubule-targeting drug and is used in the clinic for resistant or metastatic cancer. Previously, we showed that P-glycoprotein (P-gp)-overexpressing KBV20C oral cancer cells are highly resistant to HAL compared to sensitive KB cells. This qualitative study was designed to identify specific P-gp inhibitors that increase the sensitivity of highly resistant cancer cells to HAL. In order to identify functional P-gp inhibitors, HAL-treated KBV20C cells were co-treated with P-gp inhibitors, verapamil, elacridar, cyclosporine A, mitotane, piperine, fumagillin, curcumin, indomethacin, probenecid, sulindac, tesmilifene, and C-4. We then evaluated which P-gp inhibitors required a low dose to sensitize KBV20C cells to HAL. We also determined whether a low dose of a P-gp inhibitor could inhibit P-gp efflux pumping. We found that cyclosporine A sensitized HAL-treated KBV20C cells at a low dose, whereas verapamil, another first-generation P-gp inhibitor, required a dose that was nearly 10-fold higher. We also found that the natural products, piperine and mitotane, sensitized KBV20C cells to HAL co-treatment. Interestingly, we found that elacridar, a third-generation P-gp inhibitor, sensitized HAL-treated cells at a low dose. Elacridar required approximately a 500-fold lower dose than that of verapamil to exert a similar effect. All inhibitors showed P-gp inhibitory activity that correlated with sensitivity to HAL. These results suggest that highly HAL-resistant cancer cells can be sensitized with cyclosporine A or elacridar, specific P-gp inhibitors that exert their effects at a low dose. These findings provide important information regarding the sensitization of highly HAL-resistant cells with selective P-gp inhibitors and indicate that elacridar may be used to treat such highly HAL-resistant cancer cells.


B-cell subsets in the joint compartments of seropositive and seronegative rheumatoid arthritis (RA) and No-RA arthritides express memory markers and ZAP70 and characterize the aggregate pattern irrespectively of the autoantibody status.
The aim of the present study was to determine whether different subsets of B cells characterize synovial fluid (SF) or synovial tissue (ST) of seropositive or seronegative rheumatoid arthritis (RA) with respect to the peripheral blood (PB). PB, SF and ST of 14 autoantibody (AB)-positive (rheumatoid factor [RF]-IgM, RF-IgA, anti-citrullinated peptide [CCP]), 13 negative RA and 13 no-RA chronic arthritides were examined for B-cell subsets (Bm1-Bm5 and IgD-CD27 classifications), zeta-associated protein kinase-70 (ZAP70) expression on B cells and cytokine levels (interleukin [IL]-1β, tumor necrosis factor [TNF]-α, IL-6, IL-8 and monocyte chemotactic protein [MCP]-1). Synovial tissues were classified as aggregate and diffuse patterns. No differences were found in B-cell percentages or in subsets in PB and SF between AB(+) and AB(-) RA and no-RA. In both AB(+) and AB(-) RA (and no-RA), the percentage of CD19(+)/ZAP70(+) was higher in SF than in PB (AB(+): P = 0.03; AB(-): P = 0.01; no-RA: P = 0.01). Moreover, SF of both AB(+) and AB(-) RA (and no-RA) patients was characterized by a higher percentage of IgD-CD27(+) and IgD-CD27(-) B cells and lower percentage of IgD(+)CD27(-) (P < 0.05) B cells compared to PB. In SF, ZAP70 positivity is more represented in B cell CD27(+)/IgD(-)/CD38(-). The aggregate synovitis pattern was characterized by higher percentages of Bm5 cells in SF compared with the diffuse pattern (P = 0.05). These data suggest that no difference exists between AB(+) and AB(-) in B-cell subset compartmentalization. CD27(+)/IgD(-)/ZAP70(+) memory B cells accumulate preferentially in the joints of RA, suggesting a dynamic maturation of the B cells in this compartment.


Numb chin syndrome as the initial manifestation of HIV infection.
Numb chin syndrome, which consists of focal sensory loss and paresthesias of the chin, occurs with malignant diseases. We report a patient with numb chin syndrome, caused by Burkitt's lymphoma, occurring as the initial manifestation of HIV infection.


Investigating the Nature of Late-Time High-Energy GRB Emission Through Joint Fermi /Swift Observations.
We use joint observations by the Neil Gehrels Swift X-ray Telescope (XRT) and the Fermi Large Area Telescope (LAT) of gamma-ray burst (GRB) afterglows to investigate the nature of the long-lived high-energy emission observed by Fermi LAT. Joint broadband spectral modeling of XRT and LAT data reveal that LAT non-detections of bright X-ray afterglows are consistent with a cooling break in the inferred electron synchrotron spectrum below the LAT and/or XRT energy ranges. Such a break is sufficient to suppress the high-energy emission so as to be below the LAT detection threshold. By contrast, LAT-detected bursts are best fit by a synchrotron spectrum with a cooling break that lies either between or above the XRT and LAT energy ranges. We speculate that the primary difference between GRBs with LAT afterglow detections and the non-detected population may be in the type of circumstellar environment in which these bursts occur, with late-time LAT detections preferentially selecting GRBs that occur in low wind-like circumburst density profiles. Furthermore, we find no evidence of high-energy emission in the LAT-detected population significantly in excess of the flux expected from the electron synchrotron spectrum fit to the observed X-ray emission. The lack of excess emission at high energies could be due to a shocked external medium in which the energy density in the magnetic field is stronger than or comparable to that of the relativistic electrons behind the shock, precluding the production of a dominant synchrotron self-Compton (SSC) component in the LAT energy range. Alternatively, the peak of the SSC emission could be beyond the 0.1-100 GeV energy range considered for this analysis.


Pattern of childhood injury presenting at General Hospital Aliero, Nigeria.
Injuries are important causes of morbidity and mortality in childhood. Children are vulnerable to injuries of any kind. This is a prospective study of children with injuries who presented at General Hospital Aliero. Records were kept of injuries in children age fifteen years and below that occurred from February to November 2006 at General Hospital, Aliero. We aim to study the pattern of childhood injuries presenting at General Hospital Aliero, Nigeria. Most of the injuries occurred at home 31 (48.4%) and on the road 28 (43.8%). Road traffic accidents occurred when a moving vehicle or motorcycle hit children or children falling from moving trucks. Boys 39 (60.9%) were more involved in injuries than girls 25 (39.1%). Mortality occurred in three injured children. Childhood injuries occur more in the boys and commonly Parents and guardians should not leave children unattended even for a moment. Children should always be in company of an adult when outside the home. Childhood injury can lead to serious work and financial problems for families. Health promoting and injury preventive interventions should be instituted to reduce the rate of injuries and their effects on children.


"Nonclassical" secretion of annexin A2 to the lumenal side of the enterocyte brush border membrane.
Annexin A2 is a member of the annexin family of Ca(2+)-dependent lipid binding proteins and believed to be engaged in membrane transport processes in a number of cell types. In small intestinal enterocytes, we localized annexin A2 to the brush border region, where it was found mainly on the lumenal side of the microvilli, showing an apical secretion by a "nonclassical" mechanism. In addition, annexin A2 was associated with surface-connected, deep apical tubules in the apical terminal web region and with an underlying pleiomorphic, tubulo-vesicular compartment (subapical compartment/multivesicular bodies). By subcellular fractionation, the 36 kDa full-length form of annexin A2 was approximately equally distributed between the Mg(2+)-precipitated fraction (containing intracellular and basolateral membranes) and the microvillar membrane fraction. In addition, a 33 kDa molecular form of annexin A2 was seen in the latter fraction that could be generated from the full-length annexin A2 by digestion with trypsin. Taken together, the results suggest that annexin A2 acts in exocytic apical membrane trafficking and is proteolytically cleaved in situ by pancreatic proteinases once it has become externalized to the lumenal side of the brush border membrane. On the basis of its well-known membrane fusogenic properties, we propose a model for the nonclassical membrane translocation of annexin A2.


Behavior of Americium in Simulated Wounds in Nonhuman Primates.
An americium solution injected intramuscularly into several nonhuman primates (NHPs) was found to behave differently than predicted by the wound models described in the NCRP Report 156. This was because the injection was made along with a citrate solution, which is known to be more soluble than chlorides, oxides, or nitrates on which the NCRP Report was based. A multi-exponential wound model specific to the injected americium solution was developed based on the retention in the intramuscular sites. The model was coupled with the americium systemic model to interpret the urinary excretion data and assess the intake, and it was determined that the models were adequate to predict early urinary excretion in most cases but unable to predict late urinary excretion. This was attributed to the differences in the systemic handling of americium between humans and nonhuman primates. Information on the type of wounds, solubility, particle size, mass, chemical form, etc., should always be considered when performing wound dosimetry.


Determinants of Implementation of Isolation Precautions Against Infections by Multidrug-Resistant Microorganisms: A Hospital-Based, Multicenter, Observational Study.
OBJECTIVES We aimed to ascertain the factors associated with lack of isolation precautions (IP) in patients infected or colonized by third-generation cephalosporin-resistant Enterobacteriaceae (3GCR-E) and methicillin-resistant Staphylococcus aureus (MRSA) in hospital settings. DESIGN Prospective surveillance and audit of practices. SETTING The study included 4 university hospitals in Lyon, France. PARTICIPANTS All patients hospitalized between April and June in 2013 and 2015 were included. Case patients had ≥1 clinical sample positive for MRSA and/or 3GCR-E. METHODS Factors associated with the lack of IP implementation were identified using multivariate logistic regression. The incidence of MDRO infections was expressed per 10,000 patient days. RESULTS Overall, 57,222 patients accounting for 192,234 patient days of hospitalization were included, and 635 (1.1%) MDRO cases were identified. MRSA incidence was 2.5 per 10,000 patient days (95% confidence interval [95% CI], 2.1-3.0) and 3GCR-E incidence was 10.1 per 10,000 patient days (95% CI, 9.2-11.0), with no crude difference between 2013 and 2015 (P=.15 and P=.11, respectively). Among 3GCR-E, the main species were Escherichia coli (43.8%) and Klebsiella pneumoniae (31.0%). Isolation precautions were implemented in 78.5% of cases. Lack of IP implementation was independently associated with patient age, year, specialty, hospital, colonization compared with infection, and lack of medical prescription for IPs (adjusted odds ratio, 17.4; 95% CI, 8.5-35.8; P<.001). CONCLUSIONS MRSA and 3GCR-E infections and/or colonizations are frequent in healthcare settings, and IPs are implemented in most cases. When IPs are lacking, the main factor is the absence of medical prescription for IPs, underscoring the need for alerts to physicians by the microbiological laboratory and/or the infection control team. Infect Control Hosp Epidemiol 2017;38:1188-1195.


Finite element analysis of a deformable array transducer.
Deformable array transducers have previously been described to implement 2-D phase aberration correction of near-field aberrators with only a 1xN or 2xN array configuration. This transducer design combines mechanical phase correction using an actuator with electronic phase correction for a 2-D correction with significantly fewer elements than a full 2-D array. We have previously reported the fabrication and results of a 1x32 deformable array fabricated with a RAINBOW (Reduced And INternally Biased Wafer) actuator. Because of the complicated construction of deformable arrays, we propose to use finite element analysis (FEA) as a design tool for array development. In this paper, we use 2-D and 3-D FEA to model the experimental results of the deformable array as the first step toward development of a design tool. Because the deformable array combines a mechanical actuator with a medical ultrasound transducer, improvement in performance must consider both the ultrasound characterization along with the low frequency actuator characterization. For the ultrasound characterization, time domain FEA simulations of electrical vector impedance accurately predicted the measurements of single array elements. Additionally, simulations of pulse-echo sensitivity and bandwidth were also well matched to measurements. For the low frequency actuator characterization, time domain simulation of the low frequency vector impedance accurately predicted measurement and confirmed the fundamental flexure resonance of the cantilever configuration at 1.3 kHz. Frequency domain FEA included thermal processing effects and predicted actuator curvature arising during fabrication. Finally, frequency domain FEA simulations of voltage-induced displacement accurately predicted measured displacement.


[The analysis of tiapride in cadaveric material].
To optimize conditions of tiapride isolation from cadaveric organs, we compared the results of conventional methods by Stas-Otto, A.A. Vasilyeva and V.F. Kramarenko which provide tiapride isolation up to 50% and a new precise and reproducible method providing 60 +/- 2% tiapride isolation. Identification of tiapride isolated from cadaveric material was made with thin-layer chromatography and high performance liquid chromatography. The latter assay employed the method of external standard. The original techniques proposed identify and measure tiapride in hepatic samples in the presence of unidentified endogenic compounds. The techniques are rapid, selective, sensitive and reproducible.


Time to Onset of Bendamustine-associated Skin Damage Using the Spontaneous Reporting System.
Bendamustine-associated skin damage occurs frequently in Japan and can have a profound impact on health-related quality of life. To our knowledge, there are no reports on the timing of skin damage caused by bendamustine. This study assessed trends in and the time to onset of skin damage caused by bendamustine using the Japanese Adverse Drug Reaction Reporting Database (JADER). Data related to skin damage with more than five reported cases from April 2004 to March 2021 were extracted from JADER, and the relative risk of adverse events was estimated using the reporting odds ratio and 95% confidence interval. The data were analyzed for time to onset of skin damage. JADER included a total of 2,450 reports of adverse drug reactions from bendamustine. Of these, 170 skin ailments of 10 types were reported to be associated with bendamustine. Significant associations for skin damage were found for rash, herpes zoster, and infusion-related reactions. The reporting odds ratios (with 95% confidence interval) for rash, herpes zoster, and infusion-related reaction were 1.63 (1.19-2.21), 3.25 (2.20-4.78), and 7.25 (4.84-10.85), respectively. The median onset (interquartile range) of rash, herpes zoster, and infusion-related reactions caused by bendamustine were 13 (10-28), 60 (28-107), and 6 (1-28) days, respectively. A comprehensive study using a pharmacovigilance approach enabled us to identify that a rash or infusion-related reaction may be expected within 2 weeks of treatment with bendamustine and that the onset of herpes zoster occurs at a median of 2 months after treatment with bendamustine.


An Approach to Recurrent Clubfoot Deformity in Adolescents.
Even under ideal circumstances, recurrence of infantile clubfoot deformity following the Ponseti method of treatment is to be expected to occur in as many as 20% of patients. When encountered early in childhood, these recurrences are usually amenable to further casting and limited surgery. Creation of a plantigrade foot, however, becomes much more challenging when recurrences present during adolescence and early adulthood. Because of the stiffer nature of these deformities in older patients, the fact that they are often more severe because of varying lengths of neglect, and the often deleterious effects of prior intra-articular surgeries on joint health, a principled approach is recommended for both the assessment of these feet and development of an appropriate treatment plan. In doing so, the surgeon can select the combination of nonsurgical and surgical interventions that allows for as little surgery as possible to create a plantigrade foot while maintaining any motion that is present before treatment. Although no single algorithmic approach can be applied to the variety of deformities and potentially complicating factors that are encountered in treating such patients, an understanding of the utility of preoperative casting, gradual and acute corrective techniques, and the importance of identifying and mitigating deforming forces and tendon imbalance can greatly optimize outcomes.


Do arthropods feel anxious during molts?
The molting process of arthropods, chiefly controlled by ecdysteroids, is generally considered very stressful. Our previous investigations have shown that crayfish, after having experienced stressful situations, display anxiety-like behavior (ALB), characterized by aversion to light in a dark/light plus-maze (DLPM). In the present experiments, the spontaneous exploratory behavior of isolated crayfish was analyzed in a DLPM at different stages of their molt cycle. All tested animals displayed transitory aversion to light similar to ALB, before and, mostly, after molting, but not during inter-molt. Injection of ecdysteroids into inter-molt animals elicited ALB after a delay of 4 days, suggesting a long-term, possibly indirect, hormonal effect. Importantly, ecdysteroid-induced ALB was suppressed by the injection of an anxiolytic benzodiazepine. Thus, molts and their hormonal control impose internal stress on crayfish, leading to aversion behavior that has the main characteristics of anxiety. These observations are possibly generalizable to many other arthropods.


IFN-gamma-inducible protein 10 (CXCL10) contributes to airway hyperreactivity and airway inflammation in a mouse model of asthma.
Allergic asthma is an inflammatory disease of the airways characterized by eosinophilic inflammation and airway hyper-reactivity. Cytokines and chemokines specific for Th2-type inflammation predominate in asthma and in animal models of this disease. The role of Th1-type inflammatory mediators in asthma remains controversial. IFN-gamma-inducible protein 10 (IP-10; CXCL10) is an IFN-gamma-inducible chemokine that preferentially attracts activated Th1 lymphocytes. IP-10 is up-regulated in the airways of asthmatics, but its function in asthma is unclear. To investigate the role of IP-10 in allergic airway disease, we examined the expression of IP-10 in a murine model of asthma and the effects of overexpression and deletion of IP-10 in this model using IP-10-transgenic and IP-10-deficient mice. Our experiments demonstrate that IP-10 is up-regulated in the lung after allergen challenge. Mice that overexpress IP-10 in the lung exhibited significantly increased airway hyperreactivity, eosinophilia, IL-4 levels, and CD8(+) lymphocyte recruitment compared with wild-type controls. In addition, there was an increase in the percentage of IL-4-secreting T lymphocytes in the lungs of IP-10-transgenic mice. In contrast, mice deficient in IP-10 demonstrated the opposite results compared with wild-type controls, with a significant reduction in these measures of Th2-type allergic airway inflammation. Our results demonstrate that IP-10, a Th1-type chemokine, is up-regulated in allergic pulmonary inflammation and that this contributes to the airway hyperreactivity and Th2-type inflammation seen in this model of asthma.


The effects of psychological inoculation on condom use tendencies and barriers; a randomized controlled trial.
Condom use prevents the contraction of the HIV. Research shows limited effects of education on increasing condom use. Psychological inoculation (PI) has been found to be more effective in this domain, however, its mechanism is unknown. This study examined effects of PI versus education on condom use barriers and tendencies, and its relations with cognitive dissonance, using a fully automatized online system. The study was a randomized controlled trial (RCT) and included 149 students from a German University randomly assigned to PI or a control condition. An indirect condom use test (I-CUTE), a condom use barriers questionnaire, self-reported condom use, and cognitive dissonance estimations were all assessed at baseline and one-month post-intervention. PI significantly increased I-CUTE scores when participants had sexual relations. Control participants increased in self-reported condom use and on I-CUTE scores in people without sexual relations. No changes in barriers were seen in either group. The cognitive dissonance tended to be higher in PI participants as compared to control participants. PI increases I-CUTE scores compared to controls (based on effect sizes), and significantly in those with sexual relations. The role of relationship status and the mechanisms of PI should be further examined.


Determinants of Nicotine Dependence in Chronic Obstructive Pulmonary Disease.
Smoking cessation is the most powerful intervention to modify progress of chronic obstructive pulmonary disease (COPD), and nicotine dependence is one of the most important determinants of success or failure in smoking cessation. We evaluated nicotine dependence status and investigated factors associated with moderate to high nicotine dependence in patients with COPD. We included 53 current smokers with COPD in the Korean Obstructive Lung Disease II cohort enrolled between January 2014 and March 2016. Nicotine dependence was measured by using Fagerstrom test for nicotine dependence (FTND). Cognitive function was assessed by Korean version of Montreal Cognitive Assessment. The median FTND score was 3, and 32 patients (60%) had moderate to high nicotine dependence. The median smoking amount was 44 pack-years, which was not related to nicotine dependence. Multiple logistic regression analysis revealed that high education status (odds ratio, 1.286; 95% confidence interval, 1.036-1.596; p=0.023), age <70 (odds ratio, 6.407; 95% confidence interval, 1.376-29.830; p=0.018), and mild to moderate airflow obstruction (odds ratio, 6.969; 95% confidence interval, 1.388-34.998; p=0.018) were related to moderate to high nicotine dependence. Nicotine dependence does not correlate with smoking amount, but with education level, age, and severity of airflow obstruction. Physicians should provide different strategies of smoking cessation intervention for current smokers with COPD according to their education levels, age, and severity of airflow obstruction.


The "suprasellar notch," or the tuberculum sellae as seen from below: definition, features, and clinical implications from an endoscopic endonasal perspective.
The tuberculum sellae is a bony elevation ridge that lines up the anterior aspect of the sella, dividing it from the chiasmatic groove. The recent use of the endoscopic endonasal transtuberculum approach has provided surgeons with a method to reach the suprasellar area, offering a new surgical point of view somehow "opposite" of this area. The authors of this study aimed to define the tuberculum sellae as seen from the endoscopic endonasal view while also providing CT-based systematic measurements to objectively detail the anatomical features of such a structure, which was renamed the "suprasellar notch." The authors analyzed routine skull CT scans from 24 patients with no brain pathology or fractures and measured the interoptic distance at the level of the limbus sphenoidale, the chiasmatic groove sulcal length and width, and the angle of the suprasellar notch. Indeed, the suprasellar notch was defined as the angle between 2 lines, the first passing through the tuberculum sellae midpoint and perpendicular to the cribriform plate, and a second line passing between 2 points, the midpoints of the limbus sphenoidale and the tuberculum sellae. Moreover, the authors performed on 15 cadaveric heads an endoscopic endonasal transplanum transtuberculum approach with the aid of a neuronavigator to achieve a step-by-step comparison with the radiological data. The whole CT scanning set was statistically analyzed to determine the statistical interdependency of the suprasellar notch angle with the other 3 measurements, that is, the sulcal length at the midline, the interoptic distance at the optic canal entrance, and the interoptic distance at the limbus. Based on the endoscopic endonasal view and CT imaging analysis, the authors identified a certain anatomical variability and thus introduced a new classification of the suprasellar notch: Type I, angle < 118°; Type II, angle of 118°-138°; and Type III, angle > 138°. They then analyzed the surgical implications of the endoscopic endonasal approach to the suprasellar area, which could be affected by each of these structural types. The new classification identifies 3 different types of suprasellar notch and, accordingly, their surgical relevance. Above all, the authors found that the different types of suprasellar notch can affect the osteodural defect reconstruction technique, namely the positioning/wedging of the buttress in the extradural space. A precise endoscopic anatomical knowledge of the neurovascular and bony relationships--especially in cases of a less pneumatized sphenoid sinus--is crucial when approaching the anterior skull base via a transtuberculum transplanum route.


Segmented Polyurethanes and Thermoplastic Elastomers from Elemental Sulfur with Enhanced Thermomechanical Properties and Flame Retardancy.
The production of elemental sulfur from petroleum refining has created a technological opportunity to increase the valorization of elemental sulfur by the creation of high-performance sulfur based plastics with improved thermomechanical properties, elasticity and flame retardancy. We report on a synthetic polymerization methodology to prepare the first example of sulfur based segmented multi-block polyurethanes (SPUs) and thermoplastic elastomers that incorporate an appreciable amount of sulfur into the final target material. This approach applied both the inverse vulcanization of S8 with olefinic alcohols and dynamic covalent polymerizations with dienes to prepare sulfur polyols and terpolyols that were used in polymerizations with aromatic diisocyanates and short chain diols. Using these methods, a new class of high molecular weight, soluble block copolymer polyurethanes were prepared as confirmed by Size Exclusion Chromatography, NMR spectroscopy, thermal analysis, and microscopic imaging. These sulfur-based polyurethanes were readily solution processed into large area free standing films where both the tensile strength and elasticity of these materials were controlled by variation of the sulfur polyol composition. SPUs with both high tensile strength (13-24 MPa) and ductility (348 % strain at break) were prepared, along with SPU thermoplastic elastomers (578 % strain at break) which are comparable values to classical thermoplastic polyurethanes (TPUs). The incorporation of sulfur into these polyurethanes enhanced flame retardancy in comparison to classical TPUs, which points to the opportunity to impart new properties to polymeric materials as a consequence of using elemental sulfur.


Lung function decline in former smokers and low-intensity current smokers: a secondary data analysis of the NHLBI Pooled Cohorts Study.
Former smokers now outnumber current smokers in many developed countries, and current smokers are smoking fewer cigarettes per day. Some data suggest that lung function decline normalises with smoking cessation; however, mechanistic studies suggest that lung function decline could continue. We hypothesised that former smokers and low-intensity current smokers have accelerated lung function decline compared with never-smokers, including among those without prevalent lung disease. We used data on six US population-based cohorts included in the NHLBI Pooled Cohort Study. We restricted the sample to participants with valid spirometry at two or more exams. Two cohorts recruited younger adults (≥17 years), two recruited middle-aged and older adults (≥45 years), and two recruited only elderly adults (≥65 years) with examinations done between 1983 and 2014. FEV1 decline in sustained former smokers and current smokers was compared to that of never-smokers by use of mixed models adjusted for sociodemographic and anthropometric factors. Differential FEV1 decline was also evaluated according to duration of smoking cessation and cumulative (number of pack-years) and current (number of cigarettes per day) cigarette consumption. 25 352 participants (ages 17-93 years) completed 70 228 valid spirometry exams. Over a median follow-up of 7 years (IQR 3-20), FEV1 decline at the median age (57 years) was 31·01 mL per year (95% CI 30·66-31·37) in sustained never-smokers, 34·97 mL per year (34·36-35·57) in former smokers, and 39·92 mL per year (38·92-40·92) in current smokers. With adjustment, former smokers showed an accelerated FEV1 decline of 1·82 mL per year (95% CI 1·24-2·40) compared to never-smokers, which was approximately 20% of the effect estimate for current smokers (9·21 mL per year; 95% CI 8·35-10·08). Compared to never-smokers, accelerated FEV1 decline was observed in former smokers for decades after smoking cessation and in current smokers with low cumulative cigarette consumption (<10 pack-years). With respect to current cigarette consumption, the effect estimate for FEV1 decline in current smokers consuming less than five cigarettes per day (7·65 mL per year; 95% CI 6·21-9·09) was 68% of that in current smokers consuming 30 or more cigarettes per day (11·24 mL per year; 9·86-12·62), and around five times greater than in former smokers (1·57 mL per year; 1·00-2·14). Among participants without prevalent lung disease, associations were attenuated but were consistent with the main results. Former smokers and low-intensity current smokers have accelerated lung function decline compared with never-smokers. These results suggest that all levels of smoking exposure are likely to be associated with lasting and progressive lung damage. National Institutes of Health, National Heart Lung and Blood Institute, and US Environmental Protection Agency.


Acceptor specificity of 4-alpha-glucanotransferases of mammalian glycogen debranching enzymes.
Glycogen debranching enzyme (GDE) has two distinct active sites for its 4-alpha-glucanotransferase and amylo-alpha-1,6-glucosidase activities. The GDE 4-alpha-glucanotransferases of mammals show stringent donor specificity; only alpha-glucans with an alpha-1,6-linked maltotetraosyl or maltotriosyl branch function as donors of a maltotriosyl or maltosyl residue. In this study, we investigated the acceptor specificity of the 4-alpha-glucanotransferases using methyl alpha-maltooligosides, p-nitrophenyl alpha-maltooligosides, and pyridylaminated maltooligosaccharides of various sizes as the acceptor substrates, and phosphorylase limit dextrin as the donor substrate. High-performance liquid chromatography analysis of the transfer products indicated that maltotriosyl and maltosyl residues were specifically transferred from phosphorylase limit dextrin to acceptors with a maltopentaosyl residue comprising a nonreducing-end. These results suggest that the acceptor binding sites in the active sites of mammalian GDE 4-alpha-glucanotransferases are composed of tandem subsites that are geometrically complementary to five glucose residues.


Insights into Fanconi Anaemia from the structure of human FANCE.
Fanconi Anaemia (FA) is a cancer predisposition disorder characterized by spontaneous chromosome breakage and high cellular sensitivity to genotoxic agents. In response to DNA damage, a multi-subunit assembly of FA proteins, the FA core complex, monoubiquitinates the downstream FANCD2 protein. The FANCE protein plays an essential role in the FA process of DNA repair as the FANCD2-binding component of the FA core complex. Here we report a crystallographic and biological study of human FANCE. The first structure of a FA protein reveals the presence of a repeated helical motif that provides a template for the structural rationalization of other proteins defective in Fanconi Anaemia. The portion of FANCE defined by our crystallographic analysis is sufficient for interaction with FANCD2, yielding structural information into the mode of FANCD2 recruitment to the FA core complex. Disease-associated mutations disrupt the FANCE-FANCD2 interaction, providing structural insight into the molecular mechanisms of FA pathogenesis.


Lasso peptides: structure, function, biosynthesis, and engineering.
Lasso peptides are a class of ribosomally-synthesized and posttranslationally-modified natural products with diverse bioactivities. This review describes the structure and function of all known lasso peptides (as of mid-2012) and covers our current knowledge about the biosynthesis of those molecules. The isolation and characterization of lasso peptides are also covered as are bioinformatics strategies for the discovery of new lasso peptides from genomic sequence data. Several studies on the engineering of new or improved function into lasso peptides are highlighted, and unanswered questions in the field are also described.


[Responses of dorsal and ventral thalamic neurons to visual stimulation in Emys orbicularis tortoises].
Extracellular studies have been made on the background activity and reactions to visual stimuli in neurons of nucleus rotundus and nucleus suprapeduncularis of the thalamus in the tortoise E. orbicularis. Reactions of on-off type to diffuse light flashes were recorded predominantly in both of the nuclei, whereas specific reactions to movement stimuli were found in the neurons of n. rotundus. The receptive area of the neurons in both of the nuclei involves all the visual field of one or both of the eyes. The structure of the receptive area of the neurons of n. suprapeduncularis is relatively homogeneous, whereas the receptive area of the neurons in n. rotundus exhibits a zone with an increased excitability under the horizontal meridian. After a damage of tectum opticum, the heterogeneity of the receptive areas and the specific responses to movement in cells of n. rotundus disappeared, while thresholds of the reactions increased. In the neurons of n. suprapeduncularis homogeneous structure of the receptive area remained unaffected, the thresholds of the reactions being increased as well.


Expression of an immunoglobulin heavy chain gene transfected into lymphocytes.
We determined that mouse lymphoid cell lines can be transfected at high efficiencies (10-70%) by a polyoma virus shuttle vector. With this vector, we obtained expression of a cloned mouse alpha heavy chain gene transfected into cell lines representative of all stages in B-lymphocyte development, a T-cell lymphoma line, and 3T3 fibroblasts. Heavy chain gene expression in transfected light chain-producing myeloma cells occurred at levels comparable to those in IgA-secreting myeloma cells. Heavy chains produced in transfected myeloma cells were associated with light chains in membrane-bound IgA. While T-lymphoma cells and fibroblasts were transfected at similar efficiencies to B cells, significantly lower levels of alpha heavy chains were produced. This immunoglobulin gene transfection system provides a powerful approach for defining important regulatory regions in immunoglobulin genes and for identifying lymphoid cell factors involved in immunoglobulin gene expression in B-lymphocyte development.


A phylogenetic and evolutionary analysis of antimycin biosynthesis.
Streptomyces species and other Actinobacteria are ubiquitous in diverse environments worldwide and are the source of, or inspiration for, the majority of antibiotics. The genomic era has enhanced biosynthetic understanding of these valuable chemical entities and has also provided a window into the diversity and distribution of natural product biosynthetic gene clusters. Antimycin is an inhibitor of mitochondrial cytochrome c reductase and more recently was shown to inhibit Bcl-2/Bcl-XL-related anti-apoptotic proteins commonly overproduced by cancerous cells. Here we identify 73 putative antimycin biosynthetic gene clusters (BGCs) in publicly available genome sequences of Actinobacteria and classify them based on the presence or absence of cluster-situated genes antP and antQ, which encode a kynureninase and a phosphopantetheinyl transferase (PPTase), respectively. The majority of BGCs possess either both antP and antQ (L-form) or neither (S-form), while a minority of them lack either antP or antQ (IQ- or IP-form, respectively). We also evaluate the biogeographical distribution and phylogenetic relationships of antimycin producers and BGCs. We show that antimycin BGCs occur on five of the seven continents and are frequently isolated from plants and other higher organisms. We also provide evidence for two distinct phylogenetic clades of antimycin producers and gene clusters, which delineate S-form from L- and I-form BGCs. Finally, our findings suggest that the ancestral antimycin producer harboured an L-form gene cluster which was primarily propagated by vertical transmission and subsequently diversified into S-, IQ- and IP-form biosynthetic pathways.


Tibial tray rotation and posterior slope increase risk for outliers in coronal alignment.
The extensive variation in axial rotation of tibial components can lead to coronal plane malalignment. We analyzed the change in coronal alignment induced by tray malrotation. We constructed a computer model of knee arthroplasty and used a virtual cutting guide to cut the tibia at 90° to the coronal plane. The virtual guide was rotated axially (15° medial to 15° lateral) and with posterior slopes (0° to 7°). To assess the effect of axial malrotation, we measured the coronal plane alignment of a tibial tray that was axially rotated (25° internal to 15° external), as viewed on a standard anteroposterior (AP) radiograph. Axial rotation of the cutting guide induced a varus-valgus malalignment up to 1.8° (for 15° of axial rotation combined with 7° of posterior slope). Axial malrotation of tibial tray induced a substantially higher risk of coronal plane malalignment ranging from 1.9° valgus with 15° external rotation, to over 3° varus with 25° of internal rotation. Coronal alignment of the tibial cut changed by 0.07° per degree of axial rotation and 0.22° per degree of posterior slope (linear regression, R2 > 0.99). While the effect of axial malalignment has been studied, the impact on coronal alignment is not known. Our results indicate that the direction of the cutting guide and malalignment in axial rotation alter coronal plane alignment and can increase the incidence of outliers. Cite this article: Bone Joint J 2020;102-B(6 Supple A):43-48.


Mental imagery-based self-regulation: Effects on physical activity behaviour and its cognitive and affective precursors over time.
(1) Test whether a mental imagery-based self-regulation intervention increases physical activity behaviour over 90 days; (2) Examine cognitive and affective precursors of change in physical activity behaviour. A randomized control trial with participants (N = 500) randomized to one of six intervention conditions in a 3 (risk communication format: bulleted list, table, risk ladder) x 2 (mental imagery behaviour: physical activity, active control [sleep hygiene]) factorial design. After receiving personalized risk estimates via a website on a smartphone, participants listened to an audiorecording that guided them through a mental imagery activity related to improving physical activity (intervention group) or sleep hygiene behaviour (active control). Participants received text message reminders to complete the imagery for 3 weeks post-intervention, 4 weekly text surveys to assess behaviour and its cognitive and affective precursors, and a mailed survey 90 days post-baseline. Physical activity increased over 90 days by 19.5 more minutes per week (95%CI: 2.0, 37.1) in the physical activity than the active control condition. This effect was driven by participants in the risk ladder condition, who exercised 54.8 more minutes (95%CI 15.6, 94.0) in the physical activity condition than participants in the active control sleep hygiene group. Goal planning positively predicted physical activity behaviour (b = 12.2 minutes per week, p = 0.002), but self-efficacy, image clarity, and affective attitudes towards behaviours did not (p > 0.05). Mental imagery-based self-regulation interventions can increase physical activity behaviour, particularly when supported by personalized disease risk information presented in an easy-to-understand format.


31P NMR studies of intracellular pH and phosphate metabolism during cell division cycle of Saccharomyces cerevisiae.
We have analyzed changes in intracellular pH and phosphate metabolism during the cell cycle of Saccharomyces cerevisiae (NCYC 239) by using high-resolution 31P NMR spectroscopy. High-density yeast cultures (2 x 10(8) cells per ml) were arrested prior to "start" by sequential glucose deprivation, after which they synchronously replicated DNA and divided after a final glucose feeding. Oxygenation of arrested cultures in the absence of glucose led to increased levels of sugar phosphates and ATP and an increase in intracellular pH. However, these conditions did not initiate cell cycle progression, indicating that energization is not used as an intracellular signal for initiation of the cell division cycle and that the cells need exogenous carbon sources for growth. Glucose refeeding initiated an alkaline intracellular pH transient only in the synchronous cultures, showing that increased intracellular pH accompanies the traversal of start. Changes in phosphate flow and utilization also were observed in the synchronous cultures. In particular, there was increased consumption of external phosphate during DNA synthesis. When external phosphate levels were low, the cells consumed their internal polyphosphate stores. This shows that, under these conditions, polyphosphate acts as a phosphate supply.


Association of antibiotics in infancy with early childhood obesity.
Obesity in children and adults is associated with significant health burdens, making prevention a public health imperative. Infancy may be a critical period when environmental factors exert a lasting effect on the risk for obesity; identifying modifiable factors may help to reduce this risk. To assess the impact of antibiotics prescribed in infancy (ages 0-23 months) on obesity in early childhood (ages 24-59 months). We conducted a cohort study spanning 2001-2013 using electronic health records. Cox proportional hazard models were used to adjust for demographic, practice, and clinical covariates. The study spanned a network of primary care practices affiliated with the Children's Hospital of Philadelphia including both teaching clinics and private practices in urban Philadelphia, Pennsylvania, and the surrounding region. All children with annual visits at ages 0 to 23 months, as well 1 or more visits at ages 24 to 59 months, were enrolled. The cohort comprised 64,580 children. Treatment episodes for prescribed antibiotics were ascertained up to 23 months of age. Obesity outcomes were determined directly from anthropometric measurements using National Health and Nutrition Examination Survey 2000 body mass index norms. Sixty-nine percent of children were exposed to antibiotics before age 24 months, with a mean (SD) of 2.3 (1.5) episodes per child. Cumulative exposure to antibiotics was associated with later obesity (rate ratio [RR], 1.11; 95% CI, 1.02-1.21 for ≥ 4 episodes); this effect was stronger for broad-spectrum antibiotics (RR, 1.16; 95% CI, 1.06-1.29). Early exposure to broad-spectrum antibiotics was also associated with obesity (RR, 1.11; 95% CI, 1.03-1.19 at 0-5 months of age and RR, 1.09; 95% CI, 1.04-1.14 at 6-11 months of age) but narrow-spectrum drugs were not at any age or frequency. Steroid use, male sex, urban practice, public insurance, Hispanic ethnicity, and diagnosed asthma or wheezing were also predictors of obesity; common infectious diagnoses and antireflux medications were not. Repeated exposure to broad-spectrum antibiotics at ages 0 to 23 months is associated with early childhood obesity. Because common childhood infections were the most frequent diagnoses co-occurring with broad-spectrum antibiotic prescription, narrowing antibiotic selection is potentially a modifiable risk factor for childhood obesity.


Learning to trade via direct reinforcement.
We present methods for optimizing portfolios, asset allocations, and trading systems based on direct reinforcement (DR). In this approach, investment decision-making is viewed as a stochastic control problem, and strategies are discovered directly. We present an adaptive algorithm called recurrent reinforcement learning (RRL) for discovering investment policies. The need to build forecasting models is eliminated, and better trading performance is obtained. The direct reinforcement approach differs from dynamic programming and reinforcement algorithms such as TD-learning and Q-learning, which attempt to estimate a value function for the control problem. We find that the RRL direct reinforcement framework enables a simpler problem representation, avoids Bellman's curse of dimensionality and offers compelling advantages in efficiency. We demonstrate how direct reinforcement can be used to optimize risk-adjusted investment returns (including the differential Sharpe ratio), while accounting for the effects of transaction costs. In extensive simulation work using real financial data, we find that our approach based on RRL produces better trading strategies than systems utilizing Q-learning (a value function method). Real-world applications include an intra-daily currency trader and a monthly asset allocation system for the S&P 500 Stock Index and T-Bills.