Eye movements of monkeys during discrimination learning: role of visual scanning.
Four experiments were conducted on stumptailed monkeys (Macaca arctoides) to determine whether the high levels of visual scanning (shifts in fixation from one discriminative stimulus to the other) seen during discrimination learning play a necessary role in this learning. In Experiments 1 and 2, the monkeys were given a series of two-choice, dot-pattern discrimination reversal problems. Normal visual scanning before a choice response was allowed during all but the reversal trials of half of the problems. On these latter trials, the discriminative stimuli were replaced by an uninformative stimulus after the animals made more than one visual fixation on each discriminative stimulus. Thus, on these trials, the animals were limited, in terms of the information received, to the empirically determined minimum number of scans necessary to maintain high levels of performance on such problems. In both experiments, which differed primarily in the number and type of uninformative stimuli used, the rate of reversal learning was markedly retarded by the experimental condition, with the effect persisting over a long series of problems. The magnitude of the effect was unrelated to the similarity of the uninformative stimulus to the discriminative stimuli. In Experiment 3, the monkeys were given a series of discrimination problems without reversals, during half of which the experimental condition was in effect. The results were similar to those of the first two experiments. Experiment 4 was similar to the preceding experiment except that, under the experimental condition, each trial began with the uninformative stimuli, which were replaced by the discriminative stimuli when visual scanning occurred. The uninformative stimuli had no clear-cut effect on discrimination learning in this experiment. These experiments indicate that the information provided by above-minimum levels of scanning is not necessary for discrimination learning per se, but it dose appear necessary for efficient discrimination learning.


Cytocompatibility of Mats Prepared from Different Electrospun Polymer Nanofibers.
Mats of cytocompatible polymer fibers are needed as scaffolds in tissue engineering or as wound healing supports. Most recently, they have emerged as matrix-material to allow for in situ chemo- and biosensing inside intact tissue fragments or surrogates. Electrospinning of such fibers from polymer solutions provides extended options to control the structural and functional properties of the resulting fiber mats. We have prepared electrospun polymeric fiber mats from poly(lactic acid) (PLA), polystyrene (PS), and poly(vinyl pyrrolidone) (PVP) with two different fiber densities. Mats and individual fibers were characterized with respect to their dimensions, morphology, and their compatibility with human keratinocytes (HaCaT) selected as a biological model. Microscopic inspection revealed that HaCaT cells were viable on mats from all three polymers with only a negligible fraction of dead cells, similar to planar control surfaces. Growth in the presence of the fiber mats did not alter cellular metabolism (ATP, redox state) and did not induce significant production of cytokines (interleukin-6 (IL-6); monocyte chemoattractant protein-1 (MCP-1)). However, we did observe that fiber density changed the overall topography of the resulting mats and led to differences in the establishment of continuous cell sheets. In conclusion, the findings support the suitability of electrospun polymeric fiber mats made from PLA, PS, or PVP as potential biocompatible matrices for future two-dimensional (2D) or three-dimensional (3D) sensing of vital parameters from tissue in health and disease.


The Early Endocrine Stress Response in Experimental Subarachnoid Hemorrhage.
In patients with severe illness, such as aneurysmal subarachnoid hemorrhage (SAH), a physiologic stress response is triggered. This includes activation of the hypothalamic-pituitary-adrenal (HPA) axis and the sympathetic nervous system. The aim of this study was to investigate the very early responses of these systems. A porcine animal model of aneurysmal SAH was used. In this model, blood is injected slowly to the basal cisterns above the anterior skull base until the cerebral perfusion pressure is 0 mm Hg. Sampling was done from blood and urine at -10, +15, +75 and +135 minutes from time of induction of SAH. Analyses of adrenocorticotropic hormone (ACTH), cortisol, aldosterone, catecholamines and chromogranin-A were performed. Plasma ACTH, serum cortisol and plasma aldosterone increased in the samples following induction of SAH, and started to decline after 75 minutes. Urine cortisol also increased after SAH. Urine catecholamines and their metabolites were found to increase after SAH. Many samples were however below detection level, not allowing for statistical analysis. Plasma chromogranin-A peaked at 15 minutes after SAH, and thereafter decreased. The endocrine stress response after aneurysmal SAH was found to start within 15 minutes in the HPA axis with early peak values of ACTH, cortisol and aldosterone. The fact that the concentrations of the HPA axis hormones decreased 135 minutes after SAH may suggest that a similar pattern exists in SAH patients, thus making it difficult to catch these early peak values. There were also indications of early activation of the sympathetic nervous system, but the small number of valid samples made interpretation difficult.


Single-cell stochastic modelling of the action of antimicrobial peptides on bacteria.
Antimicrobial peptides (AMPs) produced by multi-cellular organisms as their immune system's defence against microbes are actively considered as natural alternatives to conventional antibiotics. Although substantial progress has been achieved in studying the AMPs, the microscopic mechanisms of their functioning remain not well understood. Here, we develop a new theoretical framework to investigate how the AMPs are able to efficiently neutralize bacteria. In our minimal theoretical model, the most relevant processes, AMPs entering into and the following inhibition of the single bacterial cell, are described stochastically. Using complementary master equations approaches, all relevant features of bacteria clearance dynamics by AMPs, such as the probability of inhibition and the mean times before the clearance, are explicitly evaluated. It is found that both processes, entering and inhibition, are equally important for the efficient functioning of AMPs. Our theoretical method naturally explains a wide spectrum of efficiencies of existing AMPs and their heterogeneity at the single-cell level. Theoretical calculations are also consistent with existing single-cell measurements. Thus, the presented theoretical approach clarifies some microscopic aspects of the action of AMPs on bacteria.


Transcriptome-wide identification and characterization of microRNAs from castor bean (Ricinus communis L.).
MicroRNAs (miRNAs) are endogenously encoded small RNAs that post-transcriptionally regulate gene expression and play essential roles in numerous developmental and physiological processes. Currently, little information on the transcriptome and tissue-specific expression of miRNAs is available in the model non-edible oilseed crop castor bean (Ricinus communis L.), one of the most important non-edible oilseed crops cultivated worldwide. Recent advances in sequencing technologies have allowed the identification of conserved and novel miRNAs in many plant species. Here, we used high-throughput sequencing technologies to identify and characterize the miRNAs in castor bean. Five small RNA libraries were constructed for deep sequencing from root tips, leaves, developing seeds (at the initial stage, seed1; and at the fast oil accumulation stage, seed2) and endosperms in castor bean. High-throughput sequencing generated a large number of sequence reads of small RNAs in this study. In total, 86 conserved miRNAs were identified, including 63 known and 23 newly identified. Sixteen miRNA isoform variants in length were found from the conserved miRNAs of castor bean. MiRNAs displayed diverse organ-specific expression levels among five libraries. Combined with criteria for miRNA annotation and a RT-PCR approach, 72 novel miRNAs and their potential precursors were annotated and 20 miRNAs newly identified were validated. In addition, new target candidates for miRNAs newly identified in this study were proposed. The current study presents the first high-throughput small RNA sequencing study performed in castor bean to identify its miRNA population. It characterizes and increases the number of miRNAs and their isoforms identified in castor bean. The miRNA expression analysis provides a foundation for understanding castor bean miRNA organ-specific expression patterns. The present study offers an expanded picture of miRNAs for castor bean and other members in the family Euphorbiaceae.


The Peripheral Peril: Injected Insulin Induces Insulin Insensitivity in Type 1 Diabetes.
Insulin resistance is an underappreciated facet of type 1 diabetes that occurs with remarkable consistency and considerable magnitude. Although therapeutic innovations are continuing to normalize dysglycemia, a sizable body of data suggests a second metabolic abnormality-iatrogenic hyperinsulinemia-principally drives insulin resistance and its consequences in this population and has not been addressed. We review this evidence to show that injecting insulin into the peripheral circulation bypasses first-pass hepatic insulin clearance, which leads to the unintended metabolic consequence of whole-body insulin resistance. We propose restructuring insulin therapy to restore the physiological insulin balance between the hepatic portal and peripheral circulations and thereby avoid the complications of life-long insulin resistance. As technology rapidly advances and our ability to ensure euglycemia improves, iatrogenic insulin resistance will become the final barrier to overcome to restore normal physiology, health, and life in type 1 diabetes.


Severe compression neuropathy following sudden onset of parkinsonian immobility.
Prolonged immobility with adoption of unusual fixed body postures may lead to compression neuropathy. Although parkinsonism has not been considered to predispose to nerve compression, we report three patients with Parkinson's disease and on-off motor fluctuations who developed severe compression neuropathy following a sudden onset off period. The stereotyped posture assumed by parkinsonian patients during an off period appears to make them especially prone to injury of the radial nerve in the upper arm and the brachial plexus.


Estrogens decrease reperfusion-associated cortical ischemic damage: an MRI analysis in a transient focal ischemia model.
Early identification of irreversible cerebral ischemia is critical in defining strategies that influence neuronal survival after stroke. We used MRI to investigate the effects of 17beta-estradiol (E2) on the temporal evolution of focal ischemia. Female rats were ovariectomized and divided into 1 of 2 groups: ovariectomy alone (OVX; n=4) or ovariectomy with estrogen replacement (OVX+E2; n=3). Both groups were then subjected to 1-hour middle cerebral artery occlusion (MCAO), with the use of a standardized endovascular monofilament model, followed by reperfusion. Sequential diffusion-weighted (DWI) and T2-weighted (T2WI) MRI were obtained during and after the MCAO. In separate groups of animals (n=5 for OVX and OVX+E2), cerebral blood flow (CBF) was measured by laser-Doppler methods before, during, and after occlusion. DWI detected similar lesion characteristics during MCAO in both groups. In the OVX group, lesion size did not change during reperfusion, but the signal intensity ratio increased early and stabilized during the latter stages. In contrast, DWI lesion size decreased during reperfusion in OVX+E2 rats by 50% to 60% (P<0.05), a size reduction almost exclusively limited to cortical regions. During MCAO, the signal intensity ratio in OVX+E2 rats was reduced compared with OVX rats. Reperfusion further attenuated the signal intensity ratio in cortical but not subcortical regions (P<0.05 versus OVX). T2WI revealed no lesions in either group during MCAO, but it detected lesion sizes similar to that of DWI during reperfusion. Furthermore, similar patterns and magnitudes of estrogen treatment-related decrease in lesion size were noted after reperfusion. T2WI demonstrated less intense signal intensity ratio changes in both groups compared with DWI. There were no differences in CBF between groups either during occlusion, early reperfusion, or 1 day after reperfusion. This study strongly suggests that estrogens selectively protect cortical tissue from ischemic damage during MCAO and that this protection is exerted during both the occlusion and reperfusion phases of ischemia and does not involve an estrogen-related change in CBF.


Implementation of paediatric vision screening in urban and rural areas in Cluj County, Romania.
In 2018 and 2019, paediatric vision screening was implemented in Cluj County, Romania, where universal paediatric vision screening does not yet exist. We report on the preparation and the first year of implementation. Objectives, target population and screening protocol were defined. In cities, children were screened by kindergarten nurses. In rural areas, kindergartens have no nurses and children were screened by family doctors' nurses, initially at the doctors' offices, later also in rural kindergartens. CME-accredited training courses and treatment pathways were organised. Implementation was assessed through on-site observations, interviews, questionnaires and analysis of screening results of referred children. Out of 12,795 eligible four- and five-year-old children, 7,876 were screened in 2018. In the cities, kindergarten nurses screened most children without difficulties. In Cluj-Napoca 1.62x the average annual birth rate was screened and in the small cities 1.64x. In the rural areas, however, nurses of family doctors screened only 0.49x the birth rate. In 51 out of 75 rural communes, no screening took place in the first year. Of 118 rural family doctors' nurses, 51 had followed the course and 26 screened children. They screened only 41 children per nurse, on average, as compared to 80 in the small cities and 100 in Cluj-Napoca. Screening at rural kindergartens met with limited success. These are attended by few children because of low population density, parents working abroad or children being kept at home in case of bad weather and road conditions. Three times fewer children were screened in rural areas as compared to urban areas. Kindergartens in rural areas are too small to employ nurses and family doctors' nurses do not have easy access to many children and have competing healthcare priorities: there are 1.5x as many family doctors in urban areas as compared to rural areas. For nationwide scaling-up of vision screening, nurses should be enabled to screen a sufficient number of children in rural areas.


Glucose levels predict hospitalization for congestive heart failure in patients at high cardiovascular risk.
Patients with diabetes mellitus (DM) are at high risk of developing congestive heart failure (CHF). However, the relationships between glucose levels and CHF in people with or without a history of DM have not been well characterized. We evaluated the associations between fasting plasma glucose and risk of hospitalization for CHF during follow-up in patients at high cardiovascular risk and without CHF enrolled in a large-scale clinical trials program. Baseline fasting plasma glucose levels were assessed in 31,546 high-risk subjects with > or = 1 coronary, peripheral, or cerebrovascular disease or DM with end-organ damage who are participating in 2 ongoing parallel trials evaluating the effects of telmisartan, ramipril, or their combination (Ongoing Telmisartan Alone and in Combination With Ramipril Global Endpoint Trial [ONTARGET]; n=25,620) and the effects of telmisartan against placebo in angiotensin-converting enzyme-intolerant patients (Telmisartan Randomized Assessment Study in ACE Intolerant Subjects With Cardiovascular Disease [TRANSCEND]; n=5926). Interim analyses blinded for randomized treatment were performed to compare baseline fasting plasma glucose with the adjusted CHF event rate at a mean follow-up of 886 days. Multivariable Cox regression models were performed, and associations were reported as hazard ratios and 95% confidence intervals. Among all subjects (mean age, 67 years; 69% men), of whom 11,708 (37%) had known DM and 1006 (3.2%) had newly diagnosed DM at baseline, 668 patients were hospitalized for CHF during follow-up. After adjustment for age and sex, a 1-mmol/L-higher fasting plasma glucose was associated with a 1.10-fold-increased risk of CHF hospitalization (95% confidence interval, 1.08 to 1.12; P<0.0001). The association persisted after adjustment for age, sex, smoking, previous myocardial infarction, hypertension, waist-to-hip ratio, creatinine, DM, and use of aspirin, beta-blockers, or statins (hazard ratio, 1.05; 95% confidence interval, 1.02 to 1.08; P<0.001). Fasting plasma glucose is an independent predictor of hospitalization for CHF in high-risk subjects. These data provide theoretical support for potential direct beneficial effects of glucose lowering in reducing the risk of CHF and suggests the need for specific studies targeted at this issue.


Changes in aortic pulse wave velocity of four thoracic aortic stent grafts in an ex vivo porcine model.
Thoracic endovascular aortic repair (TEVAR) has been shown to lead to increased aortic stiffness. The aim of this study was to investigate the effect of stent graft type and stent graft length on aortic stiffness in a controlled, experimental setting. Twenty porcine thoracic aortas were connected to a pulsatile mock loop system. Intraluminal pressure was recorded at two sites in order to measure pulse wave velocity (PWV) for each aorta: before stent graft deployment (t1); after deployment of a 100-mm long stent graft (t2); and after distal extension through deployment of a second 100-mm long stent graft (t3). Four different types of stent grafts (Conformable Gore® TAG® Device, Bolton Relay® Device, Cook Zenith Alpha™, and Medtronic Valiant®) were evaluated. For the total cohort of 20 aortas, PWV increased by a mean 0.6 m/s or 8.9% of baseline PWV after deployment of a 100-mm proximal stent graft (P<0.001), and by a mean 1.4 m/s or 23.0% of baseline PWV after distal extension of the stent graft (P<0.001). Univariable regression analysis showed a significant correlation between aortic PWV and extent of stent graft coverage, (P<0.001), but no significant effect of baseline aortic length, baseline aortic PWV, or stent graft type on the percentual increase in PWV at t2 or at t3. In this experimental set-up, aortic stiffness increased significantly after stent graft deployment with each of the four types of stent graft, with the increase in aortic stiffness depending on the extent of stent graft coverage.


Influence of pre-PTCA strategy and initial PTCA result in patients with multivessel disease: the Bypass Angioplasty Revascularization Investigation (BARI).
In PTCA patients with multivessel coronary artery disease, incomplete revascularization (IR) is the result of both pre-PTCA strategy and initial lesion outcome. The unique contribution of these components on long-term patient outcome is uncertain. From the Bypass Angioplasty Revascularization Investigation (BARI), 2047 patients who underwent first-time PTCA were evaluated. Before enrollment, all significant lesions were assessed by the PTCA operator for clinical importance and intention to dilate. Complete revascularization (CR) was defined as successful dilatation of all clinically relevant lesions. Planned CR was indicated in 65% of all patients. More lesions were intended for PTCA in these patients compared with those with planned IR (2.8 versus 2.1). Successful dilatation of all intended lesions occurred in 45% of patients with planned CR versus 56% with planned IR (P<0. 001). In multivariable analysis, planned IR (versus planned CR), initial lesions attempted (not all versus all intended lesions attempted), and initial lesion outcome (not all versus all attempted lesions successful) were unrelated to 5-year risk of cardiac death or death/myocardial infarction but were all independently related to risk of CABG. Overall, a pre-PTCA strategy of IR in BARI-like patients appears comparable to a strategy of CR except for a higher need for CABG. Whether the use of new devices may attenuate the elevated risk of CABG in patients with multivessel disease and planned IR remains to be determined.


Neuropeptides in the airways: a review.
Nerve fibers containing neuropeptide Y, vasoactive intestinal polypeptide/peptide histidine isoleucine, substance P/neurokinin A (NKA), calcitonin-gene-related peptide (CGRP), and galanin are numerous in the respiratory tract. The fibers derive from several different ganglia such as the superior cervical and stellate ganglia, the jugular-nodose ganglionic complex, small ganglia along the vagus nerve, local ganglionic formations in the tracheal wall, and dorsal root ganglia at the cervical and thoracic level. The fibers surround vascular and nonvascular smooth muscle and seromucous glands and can be seen beneath the surface epithelium. In addition, nerve fibers containing substance P/NKA and CGRP penetrate into the surface epithelium. In some instances, neuropeptides are colocalized with "classic" neurotransmitters such as norepinephrine and acetylcholine; in other cases, several peptides coexist. Many of the neuropeptides affect blood flow, smooth muscle tone, and/or seromucous secretion. Some of them may take part in reflex-mediated responses. However, the lack of specific antagonists makes it difficult to define their functional role in the respiratory tract.


Novel p-arylthio cinnamides as antagonists of leukocyte function-associated antigen-1/intracellular adhesion molecule-1 interaction. 2. Mechanism of inhibition and structure-based improvement of pharmaceutical properties.
The interaction between leukocyte function-associated antigen-1 (LFA-1) and intracellular adhesion molecule-1 (ICAM-1) has been implicated in inflammatory and immune diseases. Recently, a novel series of p-arylthio cinnamides has been described as potent antagonists of the LFA-1/ICAM-1 interaction. These compounds were found to bind to the I domain of LFA-1 using two-dimensional NMR spectroscopy of 15N-labeled LFA-1 I domain. On the basis of NOE studies between compound 1 and the I domain of LFA-1, a model of the complex was constructed. This model revealed that compound 1 does not directly inhibit ICAM-1 binding by interacting with the metal ion dependent adhesion site (MIDAS). Instead, it binds to the previously proposed I domain allosteric site (IDAS) of LFA-1 and likely modulates the activation of LFA-1 through its interaction with this regulatory site. A fragment-based NMR screening strategy was applied to identify small, more water-soluble ligands that bind to a specific region of the IDAS. When incorporated into the parent cinnamide template, the resulting analogues exhibited increased aqueous solubility and improved pharmacokinetic profiles in rats, demonstrating the power of this NMR-based screening approach for rapidly modifying high-affinity ligands.


Sequence mutations of the substrate binding pocket of stem cell factor and multidrug resistance protein ABCG2 in renal cell cancer: a possible link to treatment resistance.
ABCG2 is a multidrug cellular transport protein that is associated with resistance to certain treatments in patients, particularly anticancer treatment. The tumor-protective properties of ABCG2 expression are reported to be a feature of a subset of stem cell-like tumor cells. While protection against chemotherapy has been well analyzed, the role of ABCG2 in the treatment with tyrosine kinase inhibitors is only partially understood. Tyrosine kinase inhibitors are currently the main treatment option in irresectable renal cell carcinomas. To investigate possible underlying sequence variations in the ABCG2 gene with relevance to the functional properties of the protein, 36 patient samples were analyzed. Using sequence analysis and single-nucleotide polymorphism databases, sequence variations in the highly conserved domains of the binding pocket of ABCG2 were analyzed. The resulting variations were used for computational protein prediction algorithms to identify conformational alterations. A relevant shift from A to G at position 1376 (resulting in Y→C at 459 aa) was identified and found to be present in 8.3% of the patients. These patients are currently in follow-up after resection, thus, further analysis will reveal whether this mutation has relevance to treatment efficacy.


Effect of the peptide secondary structure on the peptide amphiphile supramolecular structure and interactions.
Bottom-up fabrication of self-assembled nanomaterials requires control over forces and interactions between building blocks. We report here on the formation and architecture of supramolecular structures constructed from two different peptide amphiphiles. Inclusion of four alanines between a 16-mer peptide and a 16 carbon long aliphatic tail resulted in a secondary structure shift of the peptide headgroups from α helices to β sheets. A concomitant shift in self-assembled morphology from nanoribbons to core-shell worm-like micelles was observed by cryogenic transmission electron microscopy (cryo-TEM) and atomic force microscopy (AFM). In the presence of divalent magnesium ions, these a priori formed supramolecular structures interacted in distinct manners, highlighting the importance of peptide amphiphile design in self-assembly.


An iterative process produces oxamniquine derivatives that kill the major species of schistosomes infecting humans.
Currently there is only one method of treatment for human schistosomiasis, the drug praziquantel. Strong selective pressure has caused a serious concern for a rise in resistance to praziquantel leading to the necessity for additional pharmaceuticals, with a distinctly different mechanism of action, to be used in combination therapy with praziquantel. Previous treatment of Schistosoma mansoni included the use of oxamniquine (OXA), a prodrug that is enzymatically activated in S. mansoni but is ineffective against S. haematobium and S. japonicum. The oxamniquine activating enzyme was identified as a S. mansoni sulfotransferase (SmSULT-OR). Structural data have allowed for directed drug development in reengineering oxamniquine to be effective against S. haematobium and S. japonicum. Guided by data from X-ray crystallographic studies and Schistosoma worm killing assays on oxamniquine, our structure-based drug design approach produced a robust SAR program that tested over 300 derivatives and identified several new lead compounds with effective worm killing in vitro. Previous studies resulted in the discovery of compound CIDD-0066790, which demonstrated broad-species activity in killing of schistosome species. As these compounds are racemic mixtures, we tested and demonstrate that the R enantiomer CIDD-007229 kills S. mansoni, S. haematobium and S. japonicum better than the parent drug (CIDD-0066790). The search for derivatives that kill better than CIDD-0066790 has resulted in a derivative (CIDD- 149830) that kills 100% of S. mansoni, S. haematobium and S. japonicum adult worms within 7 days. We hypothesize that the difference in activation and thus killing by the derivatives is due to the ability of the derivative to fit in the binding pocket of each sulfotransferase (SmSULT-OR, ShSULT-OR, SjSULT-OR) and to be efficiently sulfated. The purpose of this research is to develop a second drug to be used in conjunction with praziquantel to treat the major human species of Schistosoma. Collectively, our findings show that CIDD-00149830 and CIDD-0072229 are promising novel drugs for the treatment of human schistosomiasis and strongly support further development and in vivo testing.


Acute toxicity of commonly used forestry herbicide mixtures to Ceriodaphnia dubia and Pimephales promelas.
Because many herbicides selectively control specific species or types of vegetation, they are often applied as mixtures to achieve better control over undesirable vegetation. When herbicides are applied in forest ecosystems, streams, ponds, and other bodies of water are typically protected by buffer zones in which no herbicide is applied. However, in some landscapes, small wetlands and streams are difficult to see and avoid, thus the potential acute toxicity of herbicide mixtures to aquatic organisms is of interest, yet it has not been well-studied. We examined the acute toxicity of 23 different herbicide mixtures to Ceriodaphnia dubia and fathead minnows (Pimephales promelas) at environmentally relevant concentrations, and, where possible, characterized mixture interactions using Marking's Additive Index. Maximum exposure concentrations were equivalent to applying the maximum allowable rate for each component directly to the surface of a 6-in. deep pond with no dissipation following application. Under the conditions of this study, herbicide formulations containing Accord Concentrate (glyphosate), Arsenal AC (imazapyr), Chopper (imazapyr), Escort (metsulfuron methyl), Oust XP (sulfometuron methyl), and Velpar L (hexazinone) were not associated with appreciable acute toxicity to fathead minnows or C. dubia when used alone or in mixtures with each other and various surfactants and adjuvants. Herbicide mixtures for which Additive Indexes could be calculated exhibited primarily antagonistic or simple additive toxicity. In the few cases where synergistic toxicity was observed, the degree of synergism was slight, never exceeding approximately twice the effect estimated based on additive toxicity. Based on the results of this study, neither acute toxicity nor enhanced acute aquatic toxicity due to synergistic mixture effects appears to be a significant concern for applications of the herbicide mixtures most commonly used in forestry.


Analysis of Haemophilus influenzae serotype f isolated from three Japanese children with invasive H. influenzae infection.
In Japan, publicly subsidized Haemophilus influenzae serotype b vaccines became available in 2011; consequently, the incidence of invasive H. influenzae infection in paediatric patients of less than 5 years of age decreased dramatically. In 2013, the first case of H. influenzae serotype f (Hif) meningitis in a Japanese infant was reported, and another case of Hif meningitis in a Japanese infant was observed in 2013. We experienced a fatal paediatric case of Hif bacteraemia in 2004; therefore, we conducted an analysis of the three Hif strains isolated from these three Japanese children with invasive Hif infections. All three strains were β-lactamase-non-producing, ampicillin-sensitive strains, with MICs of 1 µg ml(-1) or less. However, one of the three strains showed slightly elevated MICs for ampicillin (1 µg ml(-1)), cefotaxime (0.25 µg ml(-1)) and meropenem (0.13 µg ml(-1)). A molecular analysis by multilocus sequence typing identified all three strains as sequence type (ST) 124, which is a predominant invasive Hif strain in many countries. SmaI-digested PFGE showed variable DNA fragmentation patterns among the strains, suggesting that some highly virulent strains have originated from a single ST124 clone and caused invasive Hif infections in Japan. Additional studies are needed to determine the factors that have led to the clonal expansion of virulent ST124 strains.


[Risk factors in atherothrombotic cerebral infarct. Are there differential risk profiles in extensive cortical infarct and lacunar infarct? A case-control study].
A study of cases and controls was performed to define the factors associated with nonembolic cerebral infarction and to compare the risk profiles of extensive cortical infarction (ECI) to those of lacunar infarction (LI). One hundred and four patients with ECI were matched with 71 patients with LI and with 90 controls. Although the initial analysis disclosed multiple factors associated with cerebral infarction, the analysis with control of the factors revealed that the only factors related with the occurrence of atherothrombotic infarction were: arterial hypertension [odds ratio (OR) = 6.8], diabetes mellitus (OR = 5.6), ischemic cardiac disease (OR = 5.5), increased hematocrit value (OR = 3.0), and atherogenic lipids in patients without arterial hypertension nor diabetes. Diabetes appeared as the only factor specifically associated with LI when compared with ECI (OR = 2.1). This study supports the following conclusions: 1) the risk profile of nonembolic cerebral infarction is concise and intelligible, 2) the possible role of arterial hypertension in the genesis of LI was not confirmed, and 3) diabetic microangiopathy may be implicated in the genesis of most of LI. Conclusions from this retrospective study should be taken as an interpretation of the results since some restriction in the sensitivity of the data analysis was deliberately introduced.


GED-0507 attenuates lung fibrosis by counteracting myofibroblast transdifferentiation in vivo and in vitro.
The development of more effective, better tolerated drug treatments for progressive pulmonary fibrosis (of which idiopathic pulmonary fibrosis is the most common and severe form) is a research priority. The peroxisome proliferator-activated receptor gamma (PPAR-γ) is a key regulator of inflammation and fibrosis and therefore represents a potential therapeutic target. However, the use of synthetic PPAR-γ agonists may be limited by their potentially severe adverse effects. In a mouse model of bleomycin (BLM)-induced pulmonary fibrosis, we have demonstrated that the non-racemic selective PPAR-γ modulator GED-0507 is able to reduce body weight loss, ameliorate clinical and histological features of pulmonary fibrosis, and increase survival rate without any safety concerns. Here, we focused on the biomolecular effects of GED-0507 on various inflammatory/fibrotic pathways. We demonstrated that preventive and therapeutic administration of GED-0507 reduced the BLM-induced mRNA expression of several markers of fibrosis, including transforming growth factor (TGF)-β, alpha-smooth muscle actin, collagen and fibronectin as well as epithelial-to-mesenchymal transition (EMT) and expression of mucin 5B. The beneficial effect of GED-0507 on pulmonary fibrosis was confirmed in vitro by its ability to control TGFβ-induced myofibroblast activation in the A549 human alveolar epithelial cell line, the MRC-5 lung fibroblast line, and primary human lung fibroblasts. Compared with the US Food and Drug Administration-approved antifibrotic drugs pirfenidone and nintedanib, GED-0507 displayed greater antifibrotic activity by controlling alveolar epithelial cell dysfunction, EMT, and extracellular matrix remodeling. In conclusion, GED-0507 demonstrated potent antifibrotic properties and might be a promising drug candidate for the treatment of pulmonary fibrosis.


Hospital separations indicate increasing need for prevention of diabetic foot complications in central Australia.
The aim of this study was to guide service provision for prevention of diabetic foot complications through the analysis of hospital separation data for those with diabetes in central Australia. We reviewed the hospital separation data for central Australia from 1992 to 1997 for adults known to be diabetic and those with diabetic foot complications. Foot complications were identified from International Classification of Diseases-9th Revision (ICD-9) codes. Additionally, we assessed the proportion of cases with diabetic foot, identified by ICD-9 coding and confirmed by record review, that were identified by Australian National-Diagnostic Related Group-Version 3 (AN-DRG-3) coding. Separations with diabetes doubled from 352 in 1992 to 796 in 1997. This represents an increase from 1232 to 2521 separations per 100,000 people over 15 years of age. Separations with foot complications increased threefold from 28 in 1992 to 90 in 1997, a rate increase from 98 to 285 per 100,000 people over 15 years of age. The proportion of diabetes separations that had foot complications remained around 10% during the 6 year period. Aboriginal people made up 89% of the individuals with foot complications and 91% of separations for diabetic foot. Foot complications were predominantly of the more acute type (90%), amenable to early intervention. The AN-DRG-3 code for diabetic foot identified only 59% (37/63 in 1997) of the separations identified by the ICD-9 codes, and admitted primarily for foot complications in Alice Springs Hospital. The known burden of hospital care for diabetes and diabetic foot complications has increased markedly in recent years. A combination of changes in prevalence, primary care utilisation, detection, hospital access or re-admission rates may underlie the observed increases. As it is very unlikely that diabetes or diabetic foot complications are being over diagnosed, or that the hospitals are over utilised, this analysis shows there is an increasingly apparent need for improved prevention of diabetic foot complications. Therefore primary health care systems should ensure that they implement evidence-based care for preventing foot complications among people with diabetes.


[Acquired dyslexias and dysgraphias under the prism of cognitive neuropsychology: a model for the Spanish language].
The present paper discusses the different clinical manifestations of acquired disorders of reading and writing from a neurocognitive viewpoint. Based on a specific functional architecture of reading and writing--a cognitive model; presented as well--the different syndromes of acquired dyslexias and dysgraphias, that have been described in the specialized literature during the last 25 years, will be reviewed. The different pathologies are distributed along three different functional axes: a plurimodal component, including the semantic system, for the description of peripheric disorders of reading and writing; a lexical block which is justified by the findings in patients with surface dyslexia/dysgraphia; and a third, sublexical component, in order to illustrate the different functional impairments in phonological dyslexia/dysgraphia. Following the description of syndromes due to selective "functional lesions", we discuss deep dyslexia/dysgraphia, a syndrome due to multiple functional lesions. All of the syndromes will be justified and discussed with respect to the different components of the functional architecture presented and are based on cases of the literature and personal observations. Concluding remarks will evaluate the new insights gained by the presented functional arquitecture in relation to other cognitive models for the analysis of reading aloud and writing to dictation of single words.


[Clinical trial of a variation of combined radiation therapy of recurrent rectal cancer in women].
The results of clinical testing of the variant of associated radiotherapy for recurrent rectal cancer in 34 female patients are reported. Distant grid gamma therapy in the regime of uneven irradiation was associated with intracavitary irradiation and gammatherapy from open fields. An average focal dose under uneven irradiation was 86 Gy (8600 rad) in intracavitary gammatherapy--30 Gy (3000 rad), in open field irradiation--30 Gy (3000 rad), the radiation stress being 1.5-2 times increased within the limits of normal tisues tolerance compared with routine variants of irradiation. No marked radiation injuries hampering the continuation of the treatment were observed in the period to follow. Among 34 patients in 8 (23.5%) the complete tumor regression was gained.


Prevalence and associated factors of antibiotic exposure during pregnancy in a large French population-based study during the 2010-19 period.
Although bacterial infections are frequent during pregnancy, the prescription of antibiotics to pregnant women represents a challenge for physicians, driven by the benefit-risk balance. To assess the extent of prenatal antibiotic exposure and its associated factors. This study included pregnancies in the National Mother-Child EPI-MERES Register 2010-19 (built from the French Healthcare Data System) regardless of outcome. Antibiotic exposure was defined as having at least one antibiotic prescription filled during pregnancy. The prevalence of pregnancies exposed to antibiotics was estimated. Univariable Poisson regression with generalized estimating equations was used to compare the number of antibiotic prescriptions filled during pregnancy and the period after pregnancy with the period 1 year before pregnancy. Multivariable Poisson regression was used to investigate factors associated with antibiotic exposure during pregnancy. Among 9 769 764 pregnancies, 3 501 294 (35.8%) were exposed to antibiotics and amoxicillin was the most common. Compared with a similar period 1 year before pregnancy, the number of filled antibiotic prescriptions was lower during pregnancy [incidence rate ratio (IRR) 0.903 (95% CI 0.902-0.905)] and during the period 1 year after pregnancy [IRR 0.880 (95% CI 0.879-0.881)]. Region of residence, deprivation index, smoking-related conditions and chronic diseases (especially chronic respiratory diseases) were associated with antibiotic exposure during pregnancy. Antibiotic prescriptions are filled less frequently during pregnancy than during the preceding year. This may be due to a more relevant benefit-risk assessment. Pregnant women living with social deprivation, those with smoking-related conditions and those with chronic diseases are more likely to fill antibiotic prescriptions.


A confocal microscopic study of the formation of ganglia in the airways of fetal pig lung.
Fetal airway smooth muscle contracts to neural stimulation from early gestation. This study aimed to document the development of the nerves and ganglia within the bronchial tree of the fetal pig lung as the structural correlates for this function. The formation of these structures during lung development (pseudoglandular stage, canalicular stage, and saccular stage) was followed through to the postnatal period, using antibodies to protein gene product 9.5, a nonspecific nerve marker; synaptic vesicle protein 2, a marker of synaptic vesicle membranes; and neurofilament, a marker of filaments in the neuronal cytoskeleton. Glial cells were stained for glial fibrillary acidic protein (GFAP) and S-100, and the airway smooth muscle for alpha-actin. Whole mounts of the bronchial tree were imaged using confocal microscopy. The formation of ganglia commences in the pseudoglandular stage with patches of neuroblasts in the wall of the epithelial tubules. These ganglionic precursors are supplied with an abundance of nerve trunks and fibers that arise from the vagus and extend to the growing tips of the airways. These trunks show profiles of Schwann cells. As the airways grow, the ganglionic precursors condense at the nerve junctions. Nerve bundles in trunks and neurons in ganglia become increasingly enveloped by GFAP-positive sheaths. From midterm onward (canalicular stage), ganglia contain cholinergic neurons. In the third trimester (saccular stage) and postnatally, ganglia further increase in size and contain mainly nerve fibers in the center. Thus, neural tissue is a dominant feature of the primordial lung, which is enveloped by nerves and ganglia through gestation into postnatal life.


Short-term Outcomes of a Laparoscopic Left Hemicolectomy for Descending Colon Cancer: Retrospective Comparison with an Open Left Hemicolectomy.
Many randomized clinical trials have been performed to treat a colorectal neoplasm with the exclusion of descending colon cancer. The aim of the present study was to investigate the difference in surgical outcomes between a laparoscopic left hemicolectomy and a conventional open left hemicolectomy for descending colon cancer. A retrospective study of ninety patients with descending colon cancer, who underwent a laparoscopic (LAP) or open left hemicolectomy (OS) between May 1998 and December 2009 at Kyungpook National University Hospital, was performed. Clinicopathological and surgical outcomes were compared between the LAP and the OS for descending colon cancer. The baseline characteristics, including age, gender, body mass index, history of prior abdominal surgical history and tumor location, were similar between the two groups. The mean operation time was 156.2 minutes for the LAP group and 223.2 minutes for the OS group (P < 0.001). Intraoperative blood loss was significantly greater in the OS group (37.5 mL vs. 80.4 mL; P = 0.039). The postoperative recovery in the LAP group was faster, as reflected by the shorter time to pass gas and the shorter hospital stay. Pathological examinations showed the surgery to be equally radical in the two groups. The median follow-up was 21 months and there were 3 distant metastases (8.5%) during follow-up in the LAP group, but no port-site or local recurrence. A laparoscopic left hemicolectomy is a technically safe and feasible procedure for treating descending colon cancer. Prospective multi-center trials are necessary to establish the LAP as the standard treatment for descending colon cancer.


Psychological side effects of breast cancer screening.
Evaluated the impact of receiving abnormal mammogram results on women's anxiety and breast cancer worries and on their breast self-examination (BSE) frequency and intentions to obtain subsequent mammograms. A telephone survey was conducted with 308 women 50 years old and older approximately 3 months following a screening mammogram. Subjects included women with suspicious abnormal mammograms, nonsuspicious abnormal mammograms, and normal mammograms. Women with suspicious abnormal mammograms exhibited significantly elevated levels of mammography-related anxiety and breast cancer worries that interfered with their moods and functioning, despite the fact that diagnostic work-ups had ruled out breast cancer. Women with moderate levels of impairment in mood or functioning were more likely to practice monthly BSE than women with either high or low levels of impairment. Breast cancer worries, perceived susceptibility to breast cancer, and physician encouragement to get mammograms all exhibited independent positive relationships to mammogram intentions.


[Functional deficits of the foot after calcaneus fracture].
The present study investigated the clinical and functional outcome of calcaneal fractures. 14 patients were evaluated at a mean follow-up of 52 months. Two groups, group A with good and group B with poor calcaneal restoration were distinguished by Böhler's angle. Ten normal subjects were used for comparison. Clinical results were evaluated with a questionnaire, radiography and range of motion measurements. Plantar pressure patterns were recorded with a capacitive pressure distribution platform. A comparison of the injured and healthy foot of all patients revealed a load shift towards lateral foot regions. A comparison of the patient groups with the controls did not reveal more pronounced changes in the poorly restored group. The results demonstrate that a disturbance of foot function persists even after long healing.


The quantification of liver anatomical changes and assessment of occupant liver injury patterns.
Liver injuries can be significant in vehicle crashes. In this study, the liver anatomy was quantified in both adult and pediatric populations as a function of gender and age. Five anatomical liver measurements were determined using CT scans of 260 normal livers. These measurements include the area and volume, and the length, width, and girth of the liver (IRB HUM00041441). To characterize geometrical shape, an inscribed sphere and circumscribed ellipsoid were fitted on the measurements. In the pediatric population the liver area and volume continuously increased with age. When normalized by patient weight, volume measurements show a decrease in volume with age, suggesting that the liver occupies a smaller proportion of the body with age. In the adult population, liver measurements varied with gender. The superior and inferior locations of the liver were also recorded with respect to the spine. The lower portion was at the L3 in small children and at L2 as children approached puberty. It stayed in that area through the 60+ group, offering more ribcage protection. Liver injury patterns were also assessed in crash occupants. Seventy-two occupants with moderate to severe (AIS 2+) liver injuries were investigated. A new methodology was presented and consisted of quantifying blood volumes. The results were compared to overall liver volume and injury scales. No clear distinction on the injury pattern was observed by age group. Liver injuries were more commonly associated with AIS 2+ thoracic injuries in adults than in children. Most injuries occurred in the right lobe.


Structural connectivity as a predictive factor for responsiveness to levetiracetam treatment in epilepsy.
To investigate whether structural connectivity or glymphatic system function is a potential predictive factor for levetiracetam (LEV) response in patients with newly diagnosed epilepsy. We enrolled patients with newly diagnosed epilepsy who were administered LEV as initial monotherapy and underwent diffusion tensor imaging (DTI) at diagnosis. We categorized the patients into drug response. We used graph theory to calculate the network measures for structural connectivity based on the DTI scans in patients with epilepsy. Additionally, we evaluated glymphatic system function by calculating the DTI analysis along the perivascular space (DTI-ALPS) index based on DTI scans. We enrolled 84 patients with epilepsy. The clinical factors and DTI-ALPS index did not differ between the groups. However, some of the structural connectivity measures significantly differ between the groups. The poor responders exhibited a higher mean clustering coefficient, global efficiency, and small-worldness index than the good responders (p = 0.003, p = 0.048, and p = 0.038, respectively). In the receiver operating characteristic curve analysis, the mean clustering coefficient exhibited the highest performance in predicting the responsiveness to LEV (area under the curve of 0.677). In the multiple logistic regression analysis, the mean clustering coefficient of the structural connectivity measures was the only significant predictor of LEV response (p = 0.014). Furthermore, in the survival analysis, the mean clustering coefficient was the only significant predictor of LEV response (p = 0.026). We demonstrated that structural connectivity is a potential predictive factor for responsiveness to LEV treatment in patients with newly diagnosed epilepsy.


Marfan syndrome, inherited aortopathies and exercise: what is the right answer?
Exercise recommendations for those who have Marfan syndrome or other genetic predisposition for thoracic aortic disease remain controversial and at times ambiguous. There are no outcomes studies to help guide recommendations. In this review, we examine the guidelines regarding exercise and inherited aortic conditions, the theoretical reasoning and circumstantial evidence that support the guidelines, as well as the knowledge gaps that continue to exist.


Variation at diabetes- and obesity-associated Loci may mirror neutral patterns of human population diversity and diabetes prevalence in India.
South Asian populations harbor a high degree of genetic diversity, due in part to demographic history. Two studies on genome-wide variation in Indian populations have shown that most Indian populations show varying degrees of admixture between ancestral north Indian and ancestral south Indian components. As a result of this structure, genetic variation in India appears to follow a geographic cline. Similarly, Indian populations seem to show detectable differences in diabetes and obesity prevalence between different geographic regions of the country. We tested the hypothesis that genetic variation at diabetes- and obesity-associated loci may be potentially related to different genetic ancestries. We genotyped 2977 individuals from 61 populations across India for 18 SNPs in genes implicated in T2D and obesity. We examined patterns of variation in allele frequency across different geographical gradients and considered state of origin and language affiliation. Our results show that most of the 18 SNPs show no significant correlation with latitude, the geographic cline reported in previous studies, or by language family. Exceptions include KCNQ1 with latitude and THADA and JAK1 with language, which suggests that genetic variation at previously ascertained diabetes-associated loci may only partly mirror geographic patterns of genome-wide diversity in Indian populations.


The link between impaired oxygen supply and cognitive decline in peripheral artery disease.
Although peripheral artery disease (PAD) primarily affects large arteries outside the brain, PAD is also associated with elevated cerebral vulnerabilities, including greater risks for brain injury (such as stroke), cognitive decline and dementia. In the present review, we aim to evaluate recent literature and extract information on potential mechanisms linking PAD and consequences on the brain. Furthermore, we suggest novel therapeutic avenues to mitigate cognitive decline and reduce risk of brain injury in patients with PAD. Various interventions, notably exercise, directly or indirectly improve systemic blood flow and oxygen supply and are effective strategies in patients with PAD or cognitive decline. Moreover, triggering protective cellular and systemic mechanisms by modulating inspired oxygen concentrations are emerging as potential novel treatment strategies. While several genetic and pharmacological approaches to modulate adaptations to hypoxia showed promising results in preclinical models of PAD, no clear benefits have yet been clinically demonstrated. We argue that genetic/pharmacological regulation of the involved adaptive systems remains challenging but that therapeutic variation of inspired oxygen levels (e.g., hypoxia conditioning) are promising future interventions to mitigate associated cognitive decline in patients with PAD.


Exposure to a 50 Hz electromagnetic field induces activation of the Epstein-Barr virus genome in latently infected human lymphoid cells.
The EBV genome in latently infected lymphoid cells offers an opportunity to follow effects on the transcriptional and translational product clearly distinguishable from those of the host cell genome. Exposure of Akata cells, a human lymphoid cell line latently infected by the EBV genome, to a 50 Hz EMF resulted in an increased number of cells expressing the virus early antigens. This finding provides additional evidence that DNA can be modulated by a magnetic field.


Gallbladder epithelial cell hydraulic water permeability and volume regulation.
The hydraulic water permeability (Lp) of the cell membranes of Necturus gallbladder epithelial cells was estimated from the rate of change of cell volume after a change in the osmolality of the bathing solution. Cell volume was calculated from computer reconstruction of light microscopic images of epithelial cells obtained by the "optical slice" technique. The tissue was mounted in a miniature Ussing chamber designed to achieve optimal optical properties, rapid bath exchange, and negligible unstirred layer thickness. The control solution contained only 80% of the normal NaCl concentration, the remainder of the osmolality was made up by mannitol, a condition that did not significantly decrease the fluid absorption rate in gallbladder sac preparations. The osmotic gradient ranged from 11.5 to 41 mosmol and was achieved by the addition or removal of mannitol from the perfusion solutions. The Lp of the apical membrane of the cell was 1.0 X 10(-3) cm/s . osmol (Posm = 0.055 cm/s) and that of the basolateral membrane was 2.2 X 10(-3) cm/s . osmol (Posm = 0.12 cm/s). These values were sufficiently high so that normal fluid absorption by Necturus gallbladder could be accomplished by a 2.4-mosmol solute gradient across the apical membrane and a 1.1-mosmol gradient across the basolateral membrane. After the initial cell shrinkage or swelling resulting from the anisotonic mucosal or serosal medium, cell volume returned rapidly toward the control value despite the fact that one bathing solution remained anisotonic. This volume regulatory response was not influenced by serosal ouabain or reduction of bath NaCl concentration to 10 mM. Complete removal of mucosal perfusate NaCl abolished volume regulation after cell shrinkage. Estimates were also made of the reflection coefficient for NaCl and urea at the apical cell membrane and of the velocity of water flow across the cytoplasm.


Calcineurin is required for Candida glabrata Pdr1 transcriptional activation.
Fluconazole is one of the most commonly used antifungals today. A result of this has been the inevitable selection of fluconazole-resistant organisms. This is an especially acute problem in the pathogenic yeast Candida glabrata. Elevated minimal inhibitory concentrations for fluconazole in C. glabrata are frequently associated with substitution mutations within the Zn2Cys6 zinc cluster-containing transcription factor-encoding gene PDR1. These mutant Pdr1 regulators drive constitutively high expression of target genes like CDR1 that encodes an ATP-binding cassette transporter thought to act as a drug efflux pump. Exposure of C. glabrata to fluconazole induced expression of both Pdr1 and CDR1, although little is known of the molecular basis underlying the upstream signals that trigger Pdr1 activation. Here, we show that the protein phosphatase calcineurin is required for fluconazole-dependent induction of Pdr1 transcriptional regulation. Calcineurin catalytic activity is required for normal Pdr1 regulation, and a hyperactive form of this phosphatase can decrease susceptibility to the echinocandin caspofungin but does not show a similar change for fluconazole susceptibility. Loss of calcineurin from strains expressing two different gain-of-function forms of Pdr1 also caused a decrease in CDR1 expression and increased fluconazole susceptibility, demonstrating that even these hyperactive Pdr1 regulatory mutants cannot bypass the requirement for calcineurin. Our data implicate calcineurin activity as a link tying azole and echinocandin susceptibility together via the control of transcription factor activity.IMPORTANCEDrug-resistant microorganisms are a problem in the treatment of all infectious diseases; this is an especially acute problem with fungi due to the existence of only three major classes of antifungal drugs, including the azole drug fluconazole. In the pathogenic yeast Candida glabrata, mutant forms of a transcription factor called Pdr1 are commonly associated with decreased fluconazole susceptibility and poor clinical outcomes. Here, we identify a protein phosphatase called calcineurin that is required for fluconazole-dependent induction of Pdr1 transcriptional activation and associated drug susceptibility. Gain-of-function mutant forms of Pdr1 still required the presence of calcineurin to confer normally decreased fluconazole susceptibility. Previous studies showed that calcineurin controls susceptibility to the echinocandin class of antifungal drugs, and our data demonstrate that this protein phosphatase is also required for normal azole drug susceptibility. Calcineurin plays a central role in susceptibility to two of the three major classes of antifungal drugs in C. glabrata.


The concerned significant others of people with gambling problems in a national representative sample in Sweden - a 1 year follow-up study.
Research into the impact of problem gambling on close social networks is scarce with the majority of studies only including help-seeking populations. To date only one study has examined concerned significant others (CSOs) from an epidemiological perspective and it did not consider gender. The aim of this study is to examine the health, social support, and financial situations of CSOs in a Swedish representative sample and to examine gender differences. A population study was conducted in Sweden in 2008/09 (n = 15,000, response rate 63%). Respondents were defined as CSOs if they reported that someone close to them currently or previously had problems with gambling. The group of CSOs was further examined in a 1-year follow up (weighted response rate 74% from the 8,165 respondents in the original sample). Comparisons were also made between those defined as CSOs only at baseline (47.7%, n = 554) and those defined as CSOs at both time points. In total, 18.2% of the population were considered CSOs, with no difference between women and men. Male and female CSOs experienced, to a large extent, similar problems including poor mental health, risky alcohol consumption, economic hardship, and arguments with those closest to them. Female CSOs reported less social support than other women and male CSOs had more legal problems and were more afraid of losing their jobs than other men. One year on, several problems remained even if some improvements were found. Both male and female CSOs reported more negative life events in the 1 year follow-up. Although some relationships are unknown, including between the CSOs and the individuals with gambling problems and the causal relationships between being a CSO and the range of associated problems, the results of this study indicate that gambling problems not only affect the gambling individual and their immediate close family but also the wider social network. A large proportion of the population can be defined as a CSO, half of whom are men. While male and female CSOs share many common problems, there are gender differences which need to be considered in prevention and treatment.


[Acute appendicitis in infants].
The analysis of the effectiveness of treatment of 12 infants with acute appendicitis was carried out. The authors distinguished three forms of the course of acute appendicitis.


Fluoro-Sorafenib (Regorafenib) effects on hepatoma cells: growth inhibition, quiescence, and recovery.
To evaluate the growth-inhibitory properties of the potent multi-kinase antagonist Regorafenib (Fluoro-Sorafenib), which was synthesized as a more potent Sorafenib, a Raf inhibitor and to determine whether similar mechanisms were involved, human hepatoma cell lines were grown in the presence or absence of Regorafanib and examined for growth inhibition. Western blots were performed for Raf targets, apoptosis, and autophagy. Regorafenib inhibited growth of human Hep3B, PLC/PRF/5, and HepG2 cells in a concentration- and time-dependent manner. Multiple signaling pathways were altered, including MAP kinases phospho-ERK and phospho-JNK and its target phospho-c-Jun. There was evidence for apoptosis by FACS, cleavage of caspases and increased Bax levels; as well as induction of autophagy, as judged by increased Beclin-1 and LC3 (II) levels. Prolonged drug exposure resulted in cell quiescence. Full growth recovery occurred after drug removal, unlike with doxorubicin chemotherapy. Regorafenib is a potent inhibitor of cell growth. Cells surviving Regorafenib treatment remain viable, but quiescent and capable of regrowth following drug removal. The reversibility of tumor cell growth suppression after drug removal may have clinical implications.


Lead optimization of melanin concentrating hormone receptor 1 antagonists with low hERG channel activity.
The discovery of small molecule melanin concentrating hormone receptor (MHCr1) antagonists as novel therapeutic agents has been widely pursued across the pharmaceutical industry. While multiple chemotypes of small molecule MCHr1 antagonists have been identified and shown to induce weight loss in rodent models of obesity, many of these lead compounds have been found to cross react with the hERG channel. This review describes efforts that led to the identification of two sub-series of MCHr1 antagonists with low affinity for the hERG channel. Ultimately, however, the modifications introduced to thwart hERG channel activity resulted in lead compounds with sub-optimal CNS behavior.


Entanglement Length Scale Separates Threading from Branching of Unknotted and Non-concatenated Ring Polymers in Melts.
Current theories on the conformation and dynamics of unknotted and non-concatenated ring polymers in melt conditions describe each ring as a tree-like double-folded object. While evidence from simulations supports this picture on a single ring level, other works show pairs of rings also thread each other, a feature overlooked in the tree theories. Here we reconcile this dichotomy using Monte Carlo simulations of the ring melts with different bending rigidities. We find that rings are double-folded (more strongly for stiffer rings) on and above the entanglement length scale, while the threadings are localized on smaller scales. The different theories disagree on the details of the tree structure, i.e., the fractal dimension of the backbone of the tree. In the stiffer melts we find an indication of a self-avoiding scaling of the backbone, while more flexible chains do not exhibit such a regime. Moreover, the theories commonly neglect threadings and assign different importance to the impact of the progressive constraint release (tube dilation) on single ring relaxation due to the motion of other rings. Despite that each threading creates only a small opening in the double-folded structure, the threading loops can be numerous and their length can exceed substantially the entanglement scale. We link the threading constraints to the divergence of the relaxation time of a ring, if the tube dilation is hindered by pinning a fraction of other rings in space. Current theories do not predict such divergence and predict faster than measured diffusion of rings, pointing at the relevance of the threading constraints in unpinned systems as well. Revision of the theories with explicit threading constraints might elucidate the validity of the conjectured existence of topological glass.


[Effects of propionic acid metabolic precursors in biotin-deprived rats].
Biotin deficiency induced in the rat, with a biotin-deficient diet but with added avidin involves clinical symptoms of deficiency as well as an important drop (80%) in propionyl-CoA carboxylase activity in the liver, heart and kidneys. But major biochemical anomalies (ketoacidosis and increased urinary elimination of propionic acid), characteristic of propionic acidaemia due to propionyl-CoA carboxylase deficiency in man, are not observed in the rat. Nevertheless, abnormal urinary elimination of methylcitrate and tiglylglycine reflects an appreciable decrease in the metabolism of propionyl-CoA in these animals. The propionyl-CoA load caused by the administration of metabolic precursors of this substance, mainly L-isoleucine, does not induce important biochemical variations except for excretion of propionylglycine.


Using workspace restrictiveness for adaptive velocity adjustment of assistive robots and upper limb exoskeletons.
Individuals with severe disabilities can benefit from assistive robotic systems (ARS) for performing activities of daily living. However, limited control interfaces are available for individuals who cannot use their hands for the control, and most of these interfaces require high effort to perform simple tasks. Therefore, autonomous and intelligent control strategies were proposed for assisting with the control in complex tasks. In this paper, we presented an autonomous and adaptive method for adjusting an assistive robot's velocity in different regions of its workspace and reducing the robot velocity where fine control is required. Two participants controlled a JACO assistive robot to grasp and lift a bottle with and without the velocity adjustment method. The task was performed 9.1% faster with velocity adjustment. Furthermore, analyzing the robot trajectory showed that the method recognized highly restrictive regions and reduced the robot end-effector velocity accordingly.Clinical relevance- The autonomous velocity adjustment method can ease the control of ARSs and improve their usability, leading to a higher quality of life for individuals with severe disabilities who can benefit from ARSs.


Interrelations between nuclear-factor kappa B activation, glial response and neuronal apoptosis in gerbil hippocampus after ischemia.
Spatial and temporal relations between transcriptional factor NF kappa B activation and glia reaction in gerbil hippocampus after transient cerebral ischemia has been studied. Activation of protein binding to NF kappa B consensus oligonucleotide was determined by electrophoretic mobility gel shift assay (EMSA) in homogenates from dorsal (DP- an equivalent of CA1 sector) and abdominal (AbP- containing CA2-4 and gyrus dentatus) parts of hippocampus. A significant activation of NF kappa B binding was observed exclusively in DP as early as 3 h after ischemia and at this time that response preceded any other morphological signs of postischemic tissue injury. This early enhancement of NF kappa B binding was followed by microglia activation visualized in CA1 pyramidal region at 24 h of recovery by histochemical staining with lectin from Ricinus communis (RCA-120). Simultaneously, only a moderate increase of immunostaining against glial fibrillary acidic protein (GFAP) was observed homogeneously in all parts of hippocampus. This uniform pattern of astrogliosis was preserved until postischemic day 3-4, when apoptotic DNA fragmentation in CA1 pyramidal neurons had been clearly documented by TUNEL staining. At this period however, continuous elevation of NF kappa B binding in DP corresponded with similar response manifested also in AbP of the hippocampus. These results evidence a preferential NF kappa B involvement in an early microglia activation in the apoptogenic CA1 sector, although its role in a later astrocytic response to ischemia could not be neglected too.


Relationship and community factors related to better mental health following child maltreatment among adolescents.
Research on factors associated with good mental health following child maltreatment is often based on unrepresentative samples and focuses on individual-level factors. To address these gaps, the present study examined the association between relationship- and community-level factors and overall mental health status among adolescents with and without a history of maltreatment in a representative sample. Data were drawn from the National Comorbidity Survey of Adolescents (NCS-A; n=10,148; data collection 2001-2004); a large, cross-sectional, nationally representative sample of adolescents aged 13-17 years from the United States. Having supportive parent and family relationships were significantly associated with good mental health (AOR ranging from 2.1 to 7.1). Positive community and school experiences were also significantly associated with good mental health (AOR ranging from 2.0 to 9.8). In most models, support from friends and siblings was not related to better mental health. Factors to be tested for efficacy in interventions targeted to adolescents with a history of child maltreatment include encouraging supportive parent and family relationships, and fostering positive community and school experiences.


Replacement of quaternary ammonium headgroups by tri-ornithine in cationic lipids for the improvement of gene delivery in vitro and in vivo.
Headgroups in cationic lipids play very important roles in determining transfection efficiency and toxicity in gene delivery. To better understand the influence of headgroups on gene delivery, a tri-peptide-based lipid was synthesized, wherein the usual quaternary ammonium was replaced by a tri-peptide. Though both the tri-peptide-based lipid (DAO3) and the quaternary ammonium-based lipid (DDCTMA) successfully mediated gene transfection, DAO3 was superior to DDCTMA in both in vitro and in vivo studies. Following their preparation into liposomes, the particle size, zeta potential, and DNA-binding capacity of the liposomes and lipoplexes were characterized to evaluate the efficiency of DAO3 compared to DDCTMA with regard to gene interactions. The expression of luciferase from pDNA mediated by DAO3 was 2-fold greater than than that with DDCTMA in Hep-2 cells, and DAO3/siRNA lipoplexes could silence about 60% luciferase in A549 cancer cells expressing firefly luciferase. DAO3/Luc-siRNA treatment exhibited 3-fold the efficiency of DDCTMA/Luc-siRNA in terms of in vivo luciferase RNAi with the bare density ratio of 0.54 at 48 h. Furthermore, DAO3 could mediate IGF-1R siRNA to inhibit tumor growth through silencing the expression of the IGF-1R protein, whereas DDCTMA showed nearly no effects. Most importantly, DAO3 had no obvious toxicity in vitro and in vivo, due to the biocompatibility of the peptide headgroups. In conclusion, these results demonstrated that the replacement of the quaternary ammonium headgroup by tri-ornithine may increase transfection efficiency and decrease toxicity.


Modelling the Predictors of Mobile Health (mHealth) Adoption among Healthcare Professionals in Low-Resource Environments.
This study was conducted with objectives to measure and validate the unified theory of the acceptance and use of technology (UTAUT) model as well as to identify the predictors of mobile health (mHealth) technology adoption among healthcare professionals in limited-resource settings. A cross-sectional survey was conducted at the six public and private hospitals in the two districts (Lodhran and Multan) of Punjab, Pakistan. The participants of the study comprised healthcare professionals (registered doctors and nurses) working in the participating hospitals. The findings of the seven-factor measurement model showed that behavioral intention (BI) to mHealth adoption is significantly influenced by performance expectancy (β = 0.504, CR = 5.064, p < 0.05) and self-concept (β = 0.860, CR = 5.968, p < 0.05) about mHealth technologies. The findings of the structural equation model (SEM) showed that the model is acceptable (χ2 (df = 259) = 3.207; p = 0.000; CFI = 0.891, IFI = 0.892, TLI = 0.874, RMSEA = 0.084). This study suggests that the adoption of mHealth can significantly help in improving people's access to quality healthcare resources and services as well as help in reducing costs and improving healthcare services. This study is significant in terms of identifying the predictors that play a determining role in the adoption of mHealth among healthcare professionals. This study presents an evidence-based model that provides an insight to policymakers, health organizations, governments, and political leaders in terms of facilitating, promoting, and implementing mHealth adoption plans in low-resource settings, which can significantly reduce health disparities and have a direct impact on health promotion.


Use of health services by children who were identified during the postpartum period as being at high risk of child abuse or neglect.
To examine the rates of hospitalizations of children identified as beingat high risk of maltreatment compared with a sociodemographically similar comparison group. retrospective study comparing a high-risk cohort with a sociodemographically matched comparison group at Yale-New Haven Hospital (New Haven, CT). One huntred fourteen children who were identified from January 1979 through December 1981 by clinicians on the postpartum ward as being at high risk of abuse or neglect and 114 sociodemographically similar comparison children who were matched according to date of birth, race, gender, and method of payment for the hospitalization. MAIN OUTCOME MEASURER: During the first 4 years of life, the number of hospitalizations, reasons for admissions, and appropriateness of days in the hospital using the Pediatric Appropriateness Evaluation Protocol. Significantly more high-risk children wer hospitalized (40% vs 22%; risk ratio 1.84; 95% confidence interval, 1.23-2.74). High-risk children were hospitalized for 649 (mean, 9.0) days versus 124 (mean, 3.8) days in comparison children. For admissions for medical problems, 28% of hospital days in the high-risk group were considered inappropriate by the Pediatric Appropriateness Evaluation Protocol versus 8% in the comparison group. There are substantial differences in the rates of hospitalizations, lengths of stay, and appropriateness of days in children identified as being at hish risk of maltreatment compared with a sociodemographically similar, non-high-risk group. Preventive programs that are aimed at reducing rates of child maltreatment in high-risk young children also should examine the program's effects on the use of hospital care.


Studies on the reconstitution of macromomycin a d auromomycin from the chromophore and protein moieties.
The chromophores extracted from macromomycin (MCR) and auromomycin (AUR) with methanol had identical ultraviolet absorption spectra, antibacterial spectra and analytical profiles in high pressure liquid chromatography. The chromophore content of AUR was about 8 times higher than that of MCR. MCR reconstituted from the chromophore and protein fraction was identical with native MCR by Sephadex G-50 chromatography, ultraviolet absorption spectrum and antibacterial spectrum. The antibacterial activity of MCR and AUR was due to the chromophore; the protein moiety had no activity. However, the protein moiety enhanced the activity of the chromophore against Gram-positive bacteria, while it suppressed the activity against Gram-negative organisms. It also protected the chromophore from heat-inactivation.


Hockey Fans in Training: A Pilot Pragmatic Randomized Controlled Trial.
Hockey Fans in Training (Hockey FIT) is a gender-sensitized weight loss and healthy lifestyle program. We investigated 1) feasibility of recruiting and retaining overweight and obese men into a pilot pragmatic randomized controlled trial and 2) potential for Hockey FIT to lead to weight loss and improvements in other outcomes at 12 wk and 12 months. Male fans of two ice hockey teams (35-65 yr; body mass index ≥28 kg·m) located in Ontario (Canada) were randomized to intervention (Hockey FIT) or comparator (wait-list control). Hockey FIT includes a 12-wk active phase (weekly, coach-led group meetings including provision of dietary information, practice of behavior change techniques, and safe exercise sessions plus incremental pedometer walking) and a 40-wk minimally supported phase (smartphone app for sustaining physical activity, private online social network, standardized e-mails, booster session/reunion). Measurement at baseline and 12 wk (both groups) and 12 months (intervention group only) included clinical outcomes (e.g., weight) and self-reported physical activity, diet, and self-rated health. Eighty men were recruited in 4 wk; trial retention was >80% at 12 wk and >75% at 12 months. At 12 wk, the intervention group lost 3.6 kg (95% confidence interval, -5.26 to -1.90 kg) more than the comparator group (P < 0.001) and maintained this weight loss to 12 months. The intervention group also demonstrated greater improvements in other clinical measures, physical activity, diet, and self-rated health at 12 wk; most sustained to 12 months. Results suggest feasible recruitment/retention of overweight and obese men in the Hockey FIT program. Results provide evidence for the potential effectiveness of Hockey FIT for weight loss and improved health in at-risk men and, thus, evidence to proceed with a definitive trial.


Peptides derived from Mycobacterium tuberculosis Rv2301 protein are involved in invasion to human epithelial cells and macrophages.
The specific function of putative cut2 protein (or CFP25), encoded by the Rv2301 gene from Mycobacterium tuberculosis H37Rv, has not been identified yet. The aim of this study was to assess some of CFP25 characteristics and its possible biological role in Mycobacterium tuberculosis H37Rv invasion process to target cells. Molecular assays indicated that the gene encoding Rv2301 is present and transcribed in M. tuberculosis complex strains. The presence of Rv2301 protein over the bacilli surface was confirmed by Western blot and immunoelectron microscopy analyses, using goats sera inoculated with synthetic peptides derived from Rv2301 protein. Receptor-ligand binding assays with carcinomic human alveolar basal epithelial cells (A549) and macrophages derived from human histolytic lymphoma monocytes (U937) allowed us to identify five high activity binding peptides (HABPs) in both cell lines, and two additional HABPs only in A549 cells. U937 HABPs binding interactions were characterized by saturation assays, finding dissociation constants (Kd) within the nanomolar range and positive cooperativity (nH>1). Inhibition assays were performed to assess the possible biological role of Rv2301 identified HABPs, finding that some of them were able to inhibit invasion at a 5 μM concentration, compared with the cytochalasin control. On the other hand, HABPs, and especially HABP 36507 located at the N-terminus of the protein, facilitated the internalization of fluorescent latex beads into A549 cells. These findings are of vital importance for the rational selection of Rv2301 HABPs, to be included as components of an antituberculosis vaccine.


Strategies for controlled synthesis of nanoparticles derived from a group of uniform materials based on organic salts.
Over the past several years, nanomaterials derived from a group of uniform materials based on organic salts (GUMBOS) have been introduced into the scientific literature involving many analytical, biological, and technological applications. In this regard, these nanoGUMBOS have been shown to display a number of unique properties including fluorescence, magnetism, tumor targeting, and optoelectronic. To date, however, little focus has been placed on developing and refining approaches for generation of size-controlled nanoGUMBOS from GUMBOS building blocks. Herein, we describe a systematic effort to define various strategies for the production of well-defined nanoGUMBOS. Specifically, we describe methods based on (i) sonochemical, (ii) microwave-assisted, (iii) cyclodextrin-assisted, and (iv) surfactant-assisted syntheses of nanoGUMBOS, evaluating the efficiency of each technique in controlling the size, sphericity, and uniformity of nanoGUMBOS produced. The effect of systematic variation in experimental parameters such as concentration, cation-to-anion ratio, as well as presence and type of template introduced for formation of nanoGUMBOS is also investigated.


Analysis of inverse-Gaussian apodized fiber Bragg grating.
Inverse-Gaussian apodized fiber Bragg gratings (IGAFBGs) are numerically studied using the transfer matrix method and fabricated by the commonly used phase-mask scanning technique in a single-step scanning process. The IGAFBG can serve as a dual-wavelength passband filter, whose wavelength spacing can be continuously tuned by introducing a tunable chirp through applying a strain gradient in principle. Also, an IGAFBG with identical dual passbands having 0.144nm wavelength spacing is experimentally achieved. We also show that an IGAFBG can act as a multipassband filter with varied free spectral ranges (FSRs), and the largest FSR variation of this IGAFBG is nearly seven times more than that in a comparable FBG pair filter. An IGAFBG with varied FSRs of approximately 16.125, approximately 12.25, approximately 8.5, and approximately 6.375GHz is fabricated. This multipassband varying-FSR IGAFBG filter can find applications in step-tunable microwave generations.


The cell of origin dictates the temporal course of neurofibromatosis-1 (Nf1) low-grade glioma formation.
Low-grade gliomas are one of the most common brain tumors in children, where they frequently form within the optic pathway (optic pathway gliomas; OPGs). Since many OPGs occur in the context of the Neurofibromatosis Type 1 (NF1) cancer predisposition syndrome, we have previously employed Nf1 genetically-engineered mouse (GEM) strains to study the pathogenesis of these low-grade glial neoplasms. In the light of the finding that human and mouse low-grade gliomas are composed of Olig2+ cells and that Olig2+ oligodendrocyte precursor cells (OPCs) give rise to murine high-grade gliomas, we sought to determine whether Olig2+ OPCs could be tumor-initiating cells for Nf1 optic glioma. Similar to the GFAP-Cre transgenic strain previously employed to generate Nf1 optic gliomas, Olig2+ cells also give rise to astrocytes in the murine optic nerve in vivo. However, in contrast to the GFAP-Cre strain where somatic Nf1 inactivation in embryonic neural progenitor/stem cells (Nf1flox/mut; GFAP-Cre mice) results in optic gliomas by 3 months of age in vivo, mice with Nf1 gene inactivation in Olig2+ OPCs (Nf1flox/mut; Olig2-Cre mice) do not form optic gliomas until 6 months of age. These distinct patterns of glioma latency do not reflect differences in the timing or brain location of somatic Nf1 loss. Instead, they most likely reflect the cell of origin, as somatic Nf1 loss in CD133+ neural progenitor/stem cells during late embryogenesis results in optic gliomas at 3 months of age. Collectively, these data demonstrate that the cell of origin dictates the time to tumorigenesis in murine optic glioma.


Therapeutic effects of bovine enterovirus infection on rabbits with experimentally induced adult T cell leukaemia.
A bovine enterovirus, MZ-468, showed cytopathic effects on cell line F-647a, which was established by coculture of human T cell lymphotropic virus type 1-transformed MT-2 cells and X-irradiated rabbit lymphocytes. Microcalorimetric assay showed that residual, viable, MZ-468-infected F-647a cells produced less heat than non-infected cells. The therapeutic effects of MZ-468 infection were examined in rabbits in which adult T cell-like leukaemia (ATL) had been induced by inoculation of F-647a cells (1 x 10(8) cells). Six newborn rabbits were separated into three groups: group A was inoculated with F-647a cells only; group B was treated with MZ-468 at the time of inoculation with cells; group C was treated with the same amount of virus 24 h after the inoculation with cells and then once every 4 days. Both of the animals in group A and one in group C died 10 and 11 days, and 22 days, respectively, after the inoculation with cells. Both rabbits in group B and one in group C survived for more than 4 months. The rabbits that died were examined pathologically; leukaemic infiltrations were found in the lungs of the group B rabbits, and in the lungs, spleens and livers of both group A rabbits. Two identical experiments produced almost the same findings. These results suggest that bovine enterovirus might be used clinically to prolong the life-span of ATL patients.


mu Opiate receptor immunoreactivity in rat central nervous system.
Immunoreactivity corresponding to the C-terminus of the rat mu opiate receptor can be detected by light microscopy in fiber- and terminal-like patterns in a number of rat brain and spinal cord regions, and in immunoreactive perikarya in several of these regions. Especially abundant fiber- and terminal-like patterns were localized to superficial layers of the spinal cord dorsal horn and nucleus caudalis of the spinal tract of the trigeminal, the nucleus of the solitary tract, nucleus ambiguous, locus coeruleus, interpeduncular nucleus, medial aspect of the lateral habenular nucleus, presumed "striasomes" of the caudate-putamen and nucleus accumbens. Moderate fiber and terminal densities were found in the ventral tegmental area, more medial aspects of the thalamus and hypothalamus, and several amygdaloid nuclei. Immunostained perikarya were prominent in the nucleus accumbens and also observed in the middle layers of the cerebral cortex, septum and diagonal band, preoptic area, medial thalamic and habenular nuclei, locus coeruleus, nucleus ambiguous, nucleus of the solitary tract, trigeminal nucleus caudalis, and spinal cord substantia gelatinosa zones. Many of these localizations correspond well with the previously-determined autoradiographic distributions of mu opiate receptor ligand binding, and with reports of mu opiate receptor immunoreactivity determined using other antisera. Electron microscopic immunohistochemical studies reveal details of the membrane distribution of the mu receptor in nucleus accumbens, caudate/putamen, locus coeruleus, and spinal cord. These results suggest largely neuronal and largely extrasynaptic distributions of mu receptors that show differential patterns of perikaryal, dendritic, and/or axonal immunostaining in different central nervous system zones. Identification of these distributions adds substantially to data identifying the cellular localization of the principal opiate receptor involved in both analgesic and addictive processes.


Protective role of IL-6 in vascular remodeling in Schistosoma pulmonary hypertension.
Schistosomiasis is one of the most common causes of pulmonary arterial hypertension worldwide, but the pathogenic mechanism by which the host inflammatory response contributes to vascular remodeling is unknown. We sought to identify signaling pathways that play protective or pathogenic roles in experimental Schistosoma-induced pulmonary vascular disease via whole-lung transcriptome analysis. Wild-type mice were experimentally exposed to Schistosoma mansoni ova by intraperitoneal sensitization followed by tail-vein augmentation, and the phenotype was assessed by right ventricular catheterization and tissue histology, as well as RNA and protein analysis. Whole-lung transcriptome analysis by microarray and RNA sequencing was performed, and RNA sequencing was analyzed according to two bioinformatics methods. Functional testing of the candidate IL-6 pathway was determined using IL-6 knockout mice and the signal transducers and activators of transcription protein-3 (STAT3) inhibitor S3I-201. Wild-type mice exposed to S. mansoni demonstrated increased right ventricular systolic pressure and thickness of the pulmonary vascular media. Whole-lung transcriptome analysis determined that the IL-6-STAT3-nuclear factor of activated T cells c2(NFATc2) pathway was up-regulated, as confirmed by PCR and the immunostaining of lung tissue from S. mansoni-exposed mice and patients who died of the disease. Mice lacking IL-6 or treated with S3I-201 developed pulmonary hypertension, associated with significant intima remodeling after exposure to S. mansoni. Whole-lung transcriptome analysis identified the up-regulation of the IL-6-STAT3-NFATc2 pathway, and IL-6 signaling was found to be protective against Schistosoma-induced intimal remodeling.


Bax targets mitochondria by distinct mechanisms before or during apoptotic cell death: a requirement for VDAC2 or Bak for efficient Bax apoptotic function.
In non-apoptotic cells, Bak constitutively resides in the mitochondrial outer membrane. In contrast, Bax is in a dynamic equilibrium between the cytosol and mitochondria, and is commonly predominant in the cytosol. In response to an apoptotic stimulus, Bax and Bak change conformation, leading to Bax accumulation at mitochondria and Bak/Bax oligomerization to form a pore in the mitochondrial outer membrane that is responsible for cell death. Using blue native-PAGE to investigate how Bax oligomerizes in the mitochondrial outer membrane, we observed that, like Bak, a proportion of Bax that constitutively resides at mitochondria associates with voltage-dependent anion channel (VDAC)2 prior to an apoptotic stimulus. During apoptosis, Bax dissociates from VDAC2 and homo-oligomerizes to form high molecular weight oligomers. In cells that lack VDAC2, constitutive mitochondrial localization of Bax and Bak was impaired, suggesting that VDAC2 has a role in Bax and Bak import to, or stability at, the mitochondrial outer membrane. However, following an apoptotic stimulus, Bak and Bax retained the ability to accumulate at VDAC2-deficient mitochondria and to mediate cell death. Silencing of Bak in VDAC2-deficient cells indicated that Bax required either VDAC2 or Bak in order to translocate to and oligomerize at the mitochondrial outer membrane to efficiently mediate apoptosis. In contrast, efficient Bak homo-oligomerization at the mitochondrial outer membrane and its pro-apoptotic function required neither VDAC2 nor Bax. Even a C-terminal mutant of Bax (S184L) that localizes to mitochondria did not constitutively target mitochondria deficient in VDAC2, but was recruited to mitochondria following an apoptotic stimulus dependent on Bak or upon over-expression of Bcl-xL. Together, our data suggest that Bax localizes to the mitochondrial outer membrane via alternate mechanisms, either constitutively via an interaction with VDAC2 or after activation via interaction with Bcl-2 family proteins.


Investigating explanations of socio-economic inequalities in health: the Dutch GLOBE study.
The GLOBE study is a prospective cohort study specifically aimed at the explanation of socio-economic inequalities in health in the Netherlands. The returns of the study are reviewed after ten years of follow-up, and the studies' contribution to the development of policy measures to reduce inequalities in health in the Netherlands are described. The study started in 1991 with a baseline postal survey (response rate 70.1% or n=18973, 15-74 years of age). Two sub-samples of respondents to this survey were subsequently interviewed in 1991 (response 79.4% and 72.3%, n=5667). Baseline data collection included measures of socio-economic position, health and possible explanatory factors. Follow-up involved repeated postal surveys and interviews, and routinely collected data on hospital admissions, cancer incidence and mortality by cause of death. Compared with higher socio-economic groups, lower socio-economic groups showed higher prevalence rates of poor self-reported health (perceived general health, health complaints, chronic conditions, disabilities), higher incidence rates of specific conditions (myocardial infarction) and higher rates of all-cause mortality. The higher prevalence of adverse material circumstances, unhealthy behaviour, adverse psychosocial characteristics, and adverse childhood circumstances in the lower socio-economic groups was important in the explanation of socio-economic inequalities in health. Socio-economic differences in health care utilization did not contribute to the explanation. The GLOBE study contributed significantly to the understanding of the explanation of socio-economic inequalities in health in the Netherlands. Study results were a main source of information in the development of policy measures aimed at the reduction of socio-economic inequalities in health in the Netherlands.


[Neuropathology of Alzheimer's disease and mild cognitive impairment].
The neuropathological hallmarks of Alzheimer s disease (AD) are the presence of senile (neuritic) plaques (NP), neurofibrillary tangles (NFT), synapse loss, and neuronal cell loss. Large neurons are more affected than small ones. AD patients can also have other non specific lesions, such as granulovacuolar degeneration, Hirano bodies, and Lewy bodies. AD patients present a loss of many cholinergic neurons, especially those located in the nucleus basalis of Meynert, as well as a decrease of choline acetyl transferase (ChAT) activity, which suggested that the core of the cognitive deficits of AD was caused by the loss of cholinergic input. Interestingly, recent studies conducted in subjects with mild cognitive impairment (MCI) (these are patients that are at risk of developing AD) have shown that these subjects can have NP and NFT in the neocortex and allocortex, with relatively normal levels of ChAT in a variety of brain regions. This suggests the structures of the cholinergic system are preserved in early stages of the disease. Scientists have made significant advances in understanding both the clinical manifestations and pathobiological manifestations of AD. However, the mechanisms of specific vulnerability of the cholinergic system to AD, as well as the initial process that leads to the complex neuropathological lesions are unknown.


Primary care for HIV-exposed and infected children: translating progress into practice.
Recent advances have changed the guidelines for diagnosing and managing pediatric human immunodeficiency virus (HIV) infection. HIV-exposed and HIV-infected children should be evaluated by, or in consultation with, pediatric HIV specialists. Primary care practitioners play a vital role in identification of infants and children at risk for HIV infection and can work collaboratively with pediatric HIV specialists to provide state-of-the-art care. With the use of perinatal zidovudine, perinatal transmission rates have been reduced to 3% to 4%, and they may be reduced even further by the use of combination antiretroviral therapy during pregnancy, viral load monitoring, and obstetric interventions. Diagnosis of HIV infection can be determined in all perinatally infected infants by 6 months of age. Combination antiretroviral therapy is the standard of care for HIV-infected children. It has become increasingly effective, but complex. Families living with HIV are affected by a number of psychosocial issues. Disclosure of HIV diagnosis to a child is an important clinical issue. As HIV-infected children grow older, medical and psychosocial issues may impact school performance. The plan of care to address specific needs of HIV-infected children should be a collaborative effort between the children, their families, the primary care team, and the multidisciplinary pediatric HIV specialty team.