The adolescent health review: test of a computerized screening tool in school-based clinics.
This study tested the viability of a stand-alone screening process in school-based health centers, and gauged its acceptance by patients and providers. The study also examined the prevalence of a variety of health risks disclosed in response to a new screening instrument and the relationship between these health risks and the stated purpose for the clinic visit. Seven school-based clinics located in six high schools and one alternative school in an urban school district participated in the study; 692 patients (83% female, 67% minority) completed the Adolescent Health Review (AHR), a multidimensional screening instrument that addressed 14 risk domains. The AHR was computerized for administration, scoring, and report generation. Females reported risk in significantly more domains than males (4.2 vs. 3.2; t = 4.5, p < .0001), including higher risk in family interaction problems, a history of physical or sexual abuse, emotional distress, suicidal behavior, marijuana use, and sexual activity. Significantly more males than females reported violent behavior. Risk rates were high regardless of stated purpose for the clinic visit. According to clinic staff, use of the AHR increased routine screening and the process was well accepted by patients and providers. Providers benefited from the opportunity to discuss risks with patients by using the printed reports to facilitate conversation and develop health care plans.


Fluorescently Labeled Virus Probes Show that Natural Virus Populations Can Control the Structure of Marine Microbial Communities.
Fluorescently stained viruses were used as probes to label, identify, and enumerate specific strains of bacteria and cyanobacteria in mixed microbial assemblages. Several marine virus isolates were fluorescently stained with YOYO-1 or POPO-1 (Molecular Probes, Inc.) and added to seawater samples that contained natural microbial communities. Cells to which the stained viruses adsorbed were easily distinguished from nonhost cells; typically, there was undetectable binding of stained viruses to natural microbial assemblages containing >10(sup6) bacteria ml(sup-1) but to which host cells were not added. Host cells that were added to natural seawater were quantified with 99% (plusmn) 2% (mean (plusmn) range) efficiency with fluorescently labeled virus probes (FLVPs). A marine bacterial isolate (strain PWH3a), tentatively identified as Vibrio natriegens, was introduced into natural microbial communities that were either supplemented with nutrients or untreated, and changes in the abundance of the isolate were monitored with FLVPs. Simultaneously, the concentrations of viruses that infected strain PWH3a were monitored by plaque assay. Following the addition of PWH3a, the concentration of viruses infecting this strain increased from undetectable levels (<1 ml(sup-1)) to 2.9 x 10(sup7) and 8.3 x 10(sup8) ml(sup-1) for the untreated and nutrient-enriched samples, respectively. The increase in viruses was associated with a collapse in populations of strain PWH3a from ca. 30 to 2% and 43 to 0.01% of the microbial communities in untreated and nutrient-enriched samples, respectively. These results clearly demonstrate that FLVPs can be used to identify and quantify specific groups of bacteria in mixed microbial communities. The data show as well that viruses which are present at low abundances in natural aquatic viral communities can control microbial community structure.


Transmission of vocational skills in the second part of careers: the effect of ICT and management changes.
This paper looks at the effect of technological and organisational changes on the probability for workers in the second part of their careers of transmitting their knowledge to other colleagues in their employing firm. We use matched employer-employee data to link changes occurred at the firm level with knowledge transmission behaviours measured at the individual-level. To control for selection bias based on differences in observable characteristics between workers employed in changing work environments and those employed in non-changing ones, we apply propensity score matching techniques. We find that ICT and management changes reduce significantly the probability for workers over 45 of transmitting their knowledge to their colleagues. Then, we analyse the role of training in mitigating this negative impact. To address issues of self-selection into training, we use propensity score matching methods and a proxy for unobservable productivity. We show that participation in a training program regarding ICT tools may help older workers restore their role of knowledge transmitters.


WIN1, a transcriptional activator of epidermal wax accumulation in Arabidopsis.
Epicuticular wax forms a layer of hydrophobic material on plant aerial organs, which constitutes a protective barrier between the plant and its environment. We report here the identification of WIN1, an Arabidopsis thaliana ethylene response factor-type transcription factor, which can activate wax deposition in overexpressing plants. We constitutively expressed WIN1 in transgenic Arabidopsis plants, and found that leaf epidermal wax accumulation was up to 4.5-fold higher in these plants than in control plants. A significant increase was also found in stems. Interestingly, approximately 50% of the additional wax could only be released by complete lipid extractions, suggesting that not all of the wax is superficial. Gene expression analysis indicated that a number of genes, such as CER1, KCS1, and CER2, which are known to be involved in wax biosynthesis, were induced in WIN1 overexpressors. This observation indicates that induction of wax accumulation in transgenic plants is probably mediated through an increase in the expression of genes encoding enzymes of the wax biosynthesis pathway.


Early Assessment of Right Ventricular Function in Systemic Lupus Erythematosus Patients using Strain and Strain Rate Imaging.
Right ventricular function is a crucial factor of the prognosis of systemic lupus erythematosus (SLE). To evaluate the right ventricular function in SLE patients with different degrees of pulmonary hypertension (PH) by strain and strain rate imaging. A total of 102 SLE patients and 30 healthy volunteers were studied between October 2015 and May 2016. Patients were divided into three groups according to pulmonary artery systolic pressure (PASP) estimated by echocardiography: group control (A); PASP ≤ 30 mmHg (group B, n = 37); PASP 30-50 mmHg (mild PH; group C, n = 34); and PASP ≥ 50 mmHg (moderate-to-severe PH; group D, n = 31). Longitudinal peak systolic strain (ε) and strain rate (SR), including systolic strain rate (SRs), early diastolic strain rate (SRe) and late diastolic strain rate (SRa) were measured in the basal, middle and apical segments of the right ventricular free wall in participants by two-dimensional speckle tracking echocardiography (2D-STE) from the apical four-chamber view. A p < 0.05 was set for statistical significance. The parameters of ε, SRs, SRe, and SRa were significantly decreased in groups C and D compared with groups A and B. The ε of each segments was significantly lower in group D than in group C, while there were no differences in SRs, SRe and SRa between groups C and D. Strain and strain rate imaging could early detect the right ventricular dysfunction in SLE patients with PH, and provide important value for clinical therapy and prognosis of these patients. (Arq Bras Cardiol. 2018; [online].ahead print, PP.0-0).


Perfusion preservation of rodent kidneys in a portable preservation device based on fluidics technology.
Technology that can implement the basic requirements for successful organ preservation in a portable configuration has yet to be realized. This work evaluates kidney preservation in a new class of portable organ preservation technology based on fluidics principles. During hypothermic pulsatile perfusion preservation (HPPP), oxygen consumption, renal vascular resistance (RVR), pH, pCO2, and perfusion pressure were measured. After 24 hr of preservation, perfusate distribution was assessed, and oxygen consumption, RVR, and glomerular filtration rate (GFR) were compared in perfused, statically stored, and freshly harvested kidneys. During HPPP, perfusion pressure was 5.8+/-3.3 mmHg with oxygen delivery to the organs in excess of 3.5 times the organ metabolic requirement. During function measurements, RVR was not statistically different in the three groups; however, both oxygen consumption and GFR in the statically stored organs were significantly lower than in HPPP stored or freshly harvested kidneys. Our findings suggest that full portability in a hypothermic perfusion preservation device seems feasible utilizing fluidics-based technology.


Characteristics and Post-Decision Attitudes of Non-Reporting Sexual Violence Victims.
Although the physical and psychological consequences of sexual violence can be severe, many victims do not report the violence to the police force. The current study examined the characteristics and the post-decisional attitude of the non-reporting sexual violence victims. In total, 287 victims of sexual violence completed an anonymous online questionnaire that assessed characteristics of the violence, whether or not the crime was reported, reasons for not reporting, and aspects that would have convinced non-reporters to report in retrospect. Eighty percent of the victims did not report the most recent sexual violence incident to the police ( n = 229). Nevertheless, 65% of the non-reporting victims ( n = 148) in retrospect would have reported to the police ("potential reporters"). Specific reasons for non-reporting and incident characteristics appear to relate to the post-decision attitude of non-reporting victims of sexual violence. "Lack of evidence" and "feelings of shame, guilt, and other emotions" appear to be more frequently mentioned by potential reporters as reasons for their decision to not report as compared with the definitive non-reporters. Likewise, being raped or sexually assaulted is predictive of potential reporting. Our findings are useful for policy makers to develop strategies to increase reporting rates of sexual violence victims.


Acclimation responses to temperature vary with vertical stratification: implications for vulnerability of soil-dwelling species to extreme temperature events.
The occurrence of summer heat waves is predicted to increase in amplitude and frequency in the near future, but the consequences of such extreme events are largely unknown, especially for belowground organisms. Soil organisms usually exhibit strong vertical stratification, resulting in more frequent exposure to extreme temperatures for surface-dwelling species than for soil-dwelling species. Therefore soil-dwelling species are expected to have poor acclimation responses to cope with temperature changes. We used five species of surface-dwelling and four species of soil-dwelling Collembola that habituate different depths in the soil. We tested for differences in tolerance to extreme temperatures after acclimation to warm and cold conditions. We also tested for differences in acclimation of the underlying physiology by looking at changes in membrane lipid composition. Chill coma recovery time, heat knockdown time and fatty acid profiles were determined after 1 week of acclimation to either 5 or 20 °C. Our results showed that surface-dwelling Collembola better maintained increased heat tolerance across acclimation temperatures, but no such response was found for cold tolerance. Concordantly, four of the five surface-dwelling Collembola showed up to fourfold changes in relative abundance of fatty acids after 1 week of acclimation, whereas none of the soil-dwelling species showed a significant adjustment in fatty acid composition. Strong physiological responses to temperature fluctuations may have become redundant in soil-dwelling species due to the relative thermal stability of their subterranean habitat. Based on the results of the four species studied, we expect that unless soil-dwelling species can temporarily retreat to avoid extreme temperatures, the predicted increase in heat waves under climatic change renders these soil-dwelling species more vulnerable to extinction than species with better physiological capabilities. Being able to act under a larger thermal range is probably costly and could reduce maximum performance at the optimal temperature.


Molecular epidemiology of chronic Pseudomonas aeruginosa airway infections in cystic fibrosis.
The molecular epidemiology of the chronic airway infections with Pseudomonas aeruginosa in individuals with cystic fibrosis (CF) was investigated by cross-sectional analysis of bacterial isolates from 51 CF centers and by longitudinal analysis of serial isolates which had been collected at the CF centers Hanover and Copenhagen since the onset of airway colonization over 30 years. Genotyping revealed that the P. aeruginosa population in CF is dominated by a few ubiquitous clones. The five most common clones retrieved from the CF host also belonged to the twenty most frequent clones in the environment and in other human disease habitats. Turnover of clones in CF airways was rare. At the Hanover clinic more than half of the patient cohort was still harbouring the initially acquired clone after twenty years of airway colonization. At the Copenhagen clinic, however, two rare clones replaced the initially acquired individual clones in all but one patient. The divergent epidemiology at the two sites is explained by their differential management of hygiene and antipseudomonal chemotherapy. Hygienic measures to prohibit patient-to-patient transmission and the modalities of antipseudomonal chemotherapy modify the epidemiology of the chronic P. aeruginosa infections in CF.


Strategy of aggressive steroid weaning and routine alendronate therapy to reduce bone loss after cardiac transplantation.
Osteoporosis is common after cardiac transplantation. The routine use of prednisolone posttransplantation is a major contributor to bone mineral loss. We sought to study the effectiveness of a strategy combining aggressive steroid weaning and routine prophylaxis with alendronate to reduce bone loss without adversely affecting posttransplantation survival. This retrospective clinical study compared 2 cohorts of patients. Group A included 28 patients who had undergone transplantation since June 1999, all of whom were prescribed alendronate (10 mg daily or 70 mg weekly). All were aggressively weaned off prednisolone with the aim of being steroid-free by 9 months posttransplantation. Only 10 of the 28 patients were on prednisolone at the time of the study. Group B was an historical control cohort of 28 posttransplant patients reviewed in a cross-sectional study in 1995. Only 2 patients were on osteoporosis prophylaxis with estrogen or vitamin D; 26 patients were on prednisolone at the time of the study. The groups were compared by dual-energy X-ray absorptiometry (DEXA) bone mineral densitometry at the femoral neck and lumbar spine at a mean of 3 years after transplantation. We compared the cumulative survival of the 2 groups. Cumulative survival posttransplantation was similar in both groups. Compared with group B, group A showed a significantly higher mean femoral Z-score (+0.3 vs -0.5, P=.01) and lumbar spine Z-score (0.0 vs -0.9, P<.02). The incidence of osteoporosis (defined by WHO criteria as T-score<or=-2.5) was lower in group A compared with group B (18% vs 29%, P=NS). A strategy of aggressive weaning of prednisolone postcardiac transplantation coupled with routine osteoporosis prophylaxis using alendronate was associated with significantly higher bone mass and a trend toward a reduced incidence of osteoporosis. This strategy was successful to reduce the use of steroid without any adverse effect on cumulative survival posttransplantation.


Constrictive endarteropathy following experimental subarachnoid hemorrhage.
Rhesus monkeys were subjected to subarachnoid hemorrhage by either the introduction of blood into the subarachnoid space or by mechanically rupturing the middle cerebral artery. Local cerebral blood flow, vascular compliance studies, and histological studies (light and electron microscope) were made of the vessels of all animals. In animals hemorrhaged via vascular rupture, subintimal swelling and thickening was minimally present by 3 days, becoming progressively more severe by one month. Such changes consisted of severe subintimal proliferation, fibrosis of the medial smooth muscle layer and interruption of the internal elastic membrane. Cells in the subintimal proliferative areas examined by electronmicroscopy were seen to have ultrastructural characteristics of smooth muscle. In the subarachnoid-injected hemorrhaged animals, there was no evidence of intimal proliferation or other vascular changes, which was also true of all control vessels. There was a decrease in wall elasticity of vessels exposed to mechanical rupture when compared to those exposed only to injected subarachnoid blood or normal vessels. Local cerebral blood flow was affected very little following the 2 methods of subarachnoid hemorrhage.


Treatment-seeking behaviour in urban Sri Lanka: trusting the state, trusting private providers.
Trust is central to good relationships between patients and health-care providers because, firstly, patient uncertainty about health conditions requires them to have confidence in a doctor's motives and decisions, and secondly, trust facilitates communication and patient focus which encourages people to utilise health services. This paper focuses on patient trust because of its effect on treatment-seeking behaviour and the treatment costs incurred by poor households. Drawing from other studies the paper distinguishes between trust based on the perceived technical competence of the provider, and on inter-personal dimensions of quality of care. Trust is also analysed at two inter-related levels: personal trust that is built through face-to-face encounters with providers; and more abstract institution-level trust. The paper applies these notions of trust to examine treatment-seeking behaviour in two poor urban communities in Colombo, Sri Lanka. Household survey data and qualitative data show that people from a range of income groups preferred to use public providers for more serious illnesses because public services were free and they trusted the technical competence of public providers at both a personal and institutional level. The data also show, however, that inter-personal quality of care was lacking in the public sector and that residents from the two communities, including a considerable minority of the poorest, preferred to use private providers for moderate acute illnesses. People were willing to pay for private services because it saved time, doctors listened and they could build better relationships with private doctors. Despite the strengths of Sri Lanka's public health sector, poor relationships act as an access barrier and push a range of income groups to the private sector. The threat to access and affordability posed by these poor relationships should be the focus of current reform debates.


Amphibian chytrid fungus and ranaviruses in the Northwest Territories, Canada.
Pathogens can cause serious declines in host species, and knowing where pathogens associated with host declines occur facilitates understanding host-pathogen ecology. Suspected drivers of global amphibian declines include infectious diseases, with 2 pathogens in particular, Batrachochytrium dendrobatidis (Bd) and ranaviruses, causing concern. We explored the host range and geographic distribution of Bd and ranaviruses in the Taiga Plains ecoregion of the Northwest Territories, Canada, in 2007 and 2008. Both pathogens were detected, greatly extending their known geographic distributions. Ranaviruses were widespread geographically, but found only in wood frogs. In contrast, Bd was found at a single site, but was detected in all 3 species of amphibians in the survey area (wood frogs, boreal chorus frogs, western toads). The presence of Bd in the Northwest Territories is not congruent with predicted distributions based on niche models, even though findings from other studies at northern latitudes are consistent with those same models. Unexpectedly, we also found evidence that swabs routinely used to collect samples for Bd screening detected fewer infections than toe clips. Our use and handling of the swabs was consistent with other studies, and the cause of the apparent lack of integrity of swabs is unknown. The ranaviruses detected in our study were confirmed to be Frog Virus 3 by sequence analysis of a diagnostic 500 bp region of the major capsid protein gene. It is unknown whether Bd or ranaviruses are recent arrivals to the Canadian north. However, the genetic analyses required to answer that question can inform larger debates about the origin of Bd in North America as well as the potential effects of climate change and industrial development on the distributions of these important amphibian pathogens.


Prenatal hormones alter antioxidant enzymes and lung histology in rats with congenital diaphragmatic hernia.
Prenatal administration of dexamethasone (Dex) and thyrotropin-releasing hormone (TRH) synergistically enhances lung maturity, but TRH suppresses the antioxidant enzyme activity. Prenatal hormonal therapy improves alveolar surfactant content and lung compliance in rats with congenital diaphragmatic hernia (CDH). In full term neonatal rats with CDH we studied the effects of prenatal Dex or Dex+TRH on antioxidant enzyme activity at birth, on survival, and on lung morphometry after 4 h of ventilation with 100% O2. CDH was induced by administration of 2,4-dichlorophenyl-p-nitro-phenylether (Nitrofen) on gestational day 10. Dex+TRH-treated CDH rats had lower activity of glutathione reductase after birth than did sham-treated CDH pups. Dex-treated and sham-treated pups had similar antioxidant enzyme activity. Hormonal treatment did not change survival during ventilation. The average airspace volume increased in Dex-treated CDH pups after ventilation, with a small synergistic effect after addition of TRH. On the basis of our findings, we speculate that prenatal administration of Dex is the best choice to improve lung maturity and airspace volume in CDH patients.


Transition States in a Protein Environment - ONIOM QM:MM Modeling of Isopenicillin N Synthesis.
To highlight the role of the protein in metal enzyme catalysis, we optimize ONIOM QM:MM transition states and intermediates for the full reaction of the nonheme iron enzyme isopenicillin N synthase (IPNS). Optimizations of transition states in large protein systems are possible using our new geometry optimizer with quadratic coupling between the QM and MM regions [Vreven, T. et al. Mol. Phys. 2006, 104, 701-704]. To highlight the effect of the metal center, results from the protein model are compared to results from an active site model containing only the metal center and coordinating residues [Lundberg, M. et al. Biochemistry 2008, 47, 1031-1042]. The analysis suggests that the main catalytic effect comes from the metal center, while the protein controls the reactivity to achieve high product specificity. As an example, hydrophobic residues align the valine substrate radical in a favorable conformation for thiazolidine ring closure and contribute to product selectivity and high stereospecificity. A low-barrier pathway for β-lactam formation is found where the proton required for heterolytic O-O bond cleavage comes directly from the valine N-H group of the substrate. The alternative mechanism, where the proton in O-O bond cleavage initially comes from an iron water ligand, can be disfavored by the electrostatic interactions with the surrounding protein. Explicit protein effects on transition states are typically 1-6 kcal/mol in the present enzyme and can be understood by considering whether the transition state involves large movements of the substrate as well as whether it involves electron transfer.


Localization and regulation of mouse pantothenate kinase 2.
Coenzyme A (CoA) biosynthesis is initiated by pantothenate kinase (PanK) and CoA levels are controlled through differential expression and feedback regulation of PanK isoforms. PanK2 is a mitochondrial protein in humans, but comparative genomics revealed that acquisition of a mitochondrial targeting signal was limited to primates. Human and mouse PanK2 possessed similar biochemical properties, with inhibition by acetyl-CoA and activation by palmitoylcarnitine. Mouse PanK2 localized in the cytosol, and the expression of PanK2 was higher in human brain compared to mouse brain. Differences in expression and subcellular localization should be considered in developing a mouse model for human PanK2 deficiency.


Determination of Electrostatic Parameters for a Polarizable Force Field Based on the Classical Drude Oscillator.
A procedure to determine the electrostatic parameters has been developed for a polarizable empirical force field based on the classical Drude oscillator model. Atomic charges and polarizabilities for a given molecule of interest were derived from restrained fitting to quantum-mechanical electrostatic potentials (ESP) calculated at the B3LYP/ cc-pVDZ or B3LYP/aug-cc-pVDZ levels on grid points located on concentric Connolly surfaces. The determination of the atomic polarizabilities requires a series of perturbed ESP maps, each one representing the electronic response of the molecule in the presence of a background charge placed on Connolly surfaces primarily along chemical bonds and lone pairs. Reference values for the partial atomic charges were taken from the CHARMM27 additive all-atom force field, and those for the polarizabilities were based on adjusted Miller's ahp atomic polarizability values. The fitted values of atomic polarizabilities were scaled to reflect the reduced polarization expected for the condensed media and/or to correct for the systematic underestimation of experimental molecular polarizabilities by B3LYP calculations. Following correction of the polarizabilities, the atomic charges were adjusted to reproduce gas-phase dipole moments. The developed scheme has been tested on a set of small molecules representing functional moieties of nucleic acids. The derived electrostatic parameters have been successfully applied in a preliminary polarizable molecular dynamics simulation of a DNA octamer in a box of water with sodium counterions. Thus, this study confirms the feasibility of the use of a polarizable force field based on a classical Drude model for simulations of biomolecules in the condensed phase.


A prospective study of endogenous serum hormone concentrations and breast cancer risk in premenopausal women on the island of Guernsey.
The associations between serum concentrations of oestradiol, progesterone, testosterone and sex hormone-binding globulin (SHBG) and risk of breast cancer in premenopausal women were investigated in a prospective study of breast cancer on the island of Guernsey. Sixty-two women diagnosed with breast cancer an average of 8 years subsequent to blood collection were matched for day of menstrual cycle, age and year of blood collection with 182 control subjects. Cases had a 12% higher mean oestradiol concentration over the whole menstrual cycle (P = 0.17) with a large difference at mid-cycle (75% higher, P = 0.04). Differences between cases and control subjects in progesterone (luteal phase), testosterone and SHBG were small and not statistically significant: luteal phase progesterone 9% lower in cases, P = 0.64; testosterone 4% higher, P = 0.57; SHBG 8% higher, P = 0.24. The small difference in oestradiol concentration could be aetiologically important, but larger prospective studies are needed.


Current antiplatelet treatment strategy in patients with diabetes mellitus.
Patients with diabetes mellitus (DM) have accelerated atherosclerosis with an increased risk for atherothrombotic cardiovascular complications. A state of high platelet reactivity and activation, hypercoagulability (prothrombotic state) and a subdued response to standard antiplatelet agents may explain high rate of adverse cardiovascular events in patients with DM. Several antithrombotic treatment strategies have been developed to control the prothrombotic state in patients with DM: dose modification of commonly used agents; use of potent agents; and addition of a third antithrombotic drug (triple therapy) to commonly prescribed dual antiplatelet therapy of aspirin and a P2Y12 inhibitor. The present review aims to provide an overview of the current knowledge on platelet abnormalities in patients with DM, focusing on the challenges and perspectives of antiplatelet treatment strategies in this population.


Stochastic simulations of pattern formation in excitable media.
We present a method for mesoscopic, dynamic Monte Carlo simulations of pattern formation in excitable reaction-diffusion systems. Using a two-level parallelization approach, our simulations cover the whole range of the parameter space, from the noise-dominated low-particle number regime to the quasi-deterministic high-particle number limit. Three qualitatively different case studies are performed that stand exemplary for the wide variety of excitable systems. We present mesoscopic stochastic simulations of the Gray-Scott model, of a simplified model for intracellular Ca2+ oscillations and, for the first time, of the Oregonator model. We achieve simulations with up to 10(10) particles. The software and the model files are freely available and researchers can use the models to reproduce our results or adapt and refine them for further exploration.


VEGF-induced neoangiogenesis is mediated by NAADP and two-pore channel-2-dependent Ca2+ signaling.
Vascular endothelial growth factor (VEGF) and its receptors VEGFR1/VEGFR2 play major roles in controlling angiogenesis, including vascularization of solid tumors. Here we describe a specific Ca(2+) signaling pathway linked to the VEGFR2 receptor subtype, controlling the critical angiogenic responses of endothelial cells (ECs) to VEGF. Key steps of this pathway are the involvement of the potent Ca(2+) mobilizing messenger, nicotinic acid adenine-dinucleotide phosphate (NAADP), and the specific engagement of the two-pore channel TPC2 subtype on acidic intracellular Ca(2+) stores, resulting in Ca(2+) release and angiogenic responses. Targeting this intracellular pathway pharmacologically using the NAADP antagonist Ned-19 or genetically using Tpcn2(-/-) mice was found to inhibit angiogenic responses to VEGF in vitro and in vivo. In human umbilical vein endothelial cells (HUVECs) Ned-19 abolished VEGF-induced Ca(2+) release, impairing phosphorylation of ERK1/2, Akt, eNOS, JNK, cell proliferation, cell migration, and capillary-like tube formation. Interestingly, Tpcn2 shRNA treatment abolished VEGF-induced Ca(2+) release and capillary-like tube formation. Importantly, in vivo VEGF-induced vessel formation in matrigel plugs in mice was abolished by Ned-19 and, most notably, failed to occur in Tpcn2(-/-) mice, but was unaffected in Tpcn1(-/-) animals. These results demonstrate that a VEGFR2/NAADP/TPC2/Ca(2+) signaling pathway is critical for VEGF-induced angiogenesis in vitro and in vivo. Given that VEGF can elicit both pro- and antiangiogenic responses depending upon the balance of signal transduction pathways activated, targeting specific VEGFR2 downstream signaling pathways could modify this balance, potentially leading to more finely tailored therapeutic strategies.


Population Pharmacokinetics of Trimethoprim-Sulfamethoxazole in Infants and Children.
Trimethoprim (TMP)-sulfamethoxazole (SMX) is used to treat various types of infections, including community-acquired methicillin-resistant Staphylococcus aureus (CA-MRSA) and Pneumocystis jirovecii infections in children. Pharmacokinetic (PK) data for infants and children are limited, and the optimal dosing is not known. We performed a multicenter, prospective PK study of TMP-SMX in infants and children. Separate population PK models were developed for TMP and SMX administered by the enteral route using nonlinear mixed-effects modeling. Optimal dosing was determined on the basis of the matching adult TMP exposure and attainment of the surrogate pharmacodynamic (PD) target for efficacy, a free TMP concentration above the MIC over 50% of the dosing interval. Data for a total of 153 subjects (240 samples for PK analysis) with a median postnatal age of 8 years (range, 0.1 to 20 years) contributed to the analysis for both drugs. A one-compartment model with first-order absorption and elimination characterized the TMP and SMX PK data well. Weight was included in the base model for clearance (CL/F) and volume of distribution (V/F). Both TMP and SMX CL/F increased with age. In addition, TMP and SMX CL/F were inversely related to the serum creatinine and albumin concentrations, respectively. The exposure achieved in children after oral administration of TMP-SMX at 8/40 mg/kg of body weight/day divided into administration every 12 h matched the exposure achieved in adults after administration of TMP-SMX at 320/1,600 mg/day divided into administration every 12 h and achieved the PD target for bacteria with an MIC of 0.5 mg/liter in >90% of infants and children. The exposure achieved in children after oral administration of TMP-SMX at 12/60 and 15/75 mg/kg/day divided into administration every 12 h matched the exposure achieved in adults after administration of TMP-SMX at 640/3,200 mg/day divided into administration every 12 h in subjects 6 to <21 years and 0 to <6 years of age, respectively, and was optimal for bacteria with an MIC of up to 1 mg/liter.


The effect of uric acid on outdoor copper and bronze.
Bird droppings are often quoted as a decay agent for outdoor goods, in particular buildings and statues. Undoubtedly, they represent one of the major causes of aesthetic damage on outdoor materials, but the real chemical damage they are able to induce, in particular on metals, is not so well studied. This work focused on the short term role of uric acid, the main constituent of bird urine, with respect to copper, which make such an important contribution to architectural elements of buildings and outdoor sculpture. Preliminary results of laboratory tests and analyses on real exposed samples showed that uric acid chemically affects copper and bronzes: the surface of the metal is modified and copper urates formed. Also natural patina, formed on statues and roof, react with uric acid, even if it seems to afford some protection toward bird droppings. In general, experimental results confirm that the potential chemical damage by bird droppings is significant when considering external cultural heritage such as statues, metal monuments and buildings with historic copper roofs.


A systems biology approach reveals the dose- and time-dependent effect of primary human airway epithelium tissue culture after exposure to cigarette smoke in vitro.
To establish a relevant in vitro model for systems toxicology-based mechanistic assessment of environmental stressors such as cigarette smoke (CS), we exposed human organotypic bronchial epithelial tissue cultures at the air liquid interface (ALI) to various CS doses. Previously, we compared in vitro gene expression changes with published human airway epithelia in vivo data to assess their similarities. Here, we present a follow-up evaluation of these in vitro transcriptomics data, using complementary computational approaches and an integrated mRNA-microRNA (miRNA) analysis. The main cellular pathways perturbed by CS exposure were related to stress responses (oxidative stress and xenobiotic metabolism), inflammation (inhibition of nuclear factor-κB and the interferon gamma-dependent pathway), and proliferation/differentiation. Within post-exposure periods up to 48 hours, a transient kinetic response was observed at lower CS doses, whereas higher doses resulted in more sustained responses. In conclusion, this systems toxicology approach has potential for product testing according to "21st Century Toxicology".


Upper eyelid postseptal weight placement for treatment of paralytic lagophthalmos.
Left untreated, paralytic lagophthalmos may result in corneal dryness, ulcerations, and subsequent blindness. The most common nondynamic surgical solution is upper eyelid weight placement in a superficial, pretarsal pocket, carrying the risk of visibility, extrusion, and entropion. The authors present a technique useful in patients presenting with either primary symptoms of corneal exposure or complications of previous implants that is equally efficacious, with a potentially decreased risk of complications, in which the weight is inserted into a deeper, postseptal position but requires use of a heavier weight. Nineteen patients, 15 primary and four secondary, treated between the years 2008 and 2012 with the postseptal approach, were evaluated for demographics, cause, resolution of primary symptoms, and complications. The average weight difference between that measured in the clinic versus weight used in surgery in the primary group and the average weight difference between that inserted in previous surgery versus weight placed in corrective surgery were calculated. The average weight difference was 0.213 g (range, 0 to 0.4 g) in the primary group and 0.2 g (range, 0 to 0.4 g) in the revision group. Symptoms resolved in all but one patient, neither revisions nor complications were noted, and the longest follow-up was 4 years. The retroseptal technique has proven to be safe, reproducible, and very useful in both primary and secondary upper eyelid surgery for paralytic lagophthalmos. It successfully addresses several common problems encountered with more superficial weight placement, including visibility, exposure, and entropion, but often necessitates use of a heavier weight. Therapeutic, IV.


A computational fluid dynamics of 'clap and fling' in the smallest insects.
In this paper, we have used the immersed boundary method to solve the two-dimensional Navier-Stokes equations for two immersed wings performing an idealized 'clap and fling' stroke and a 'fling' half-stroke. We calculated lift coefficients as functions of time per wing for a range of Reynolds numbers (Re) between 8 and 128. We also calculated the instantaneous streamlines around each wing throughout the stroke cycle and related the changes in lift to the relative strength and position of the leading and trailing edge vortices. Our results show that lift generation per wing during the 'clap and fling' of two wings when compared to the average lift produced by one wing with the same motion falls into two distinct patterns. For Re=64 and higher, lift is initially enhanced during the rotation of two wings when lift coefficients are compared to the case of one wing. Lift coefficients after fling and during the translational part of the stroke oscillate as the leading and trailing edge vortices are alternately shed. In addition, the lift coefficients are not substantially greater in the two-winged case than in the one-winged case. This differs from three-dimensional insect flight where the leading edge vortices remain attached to the wing throughout each half-stroke. For Re=32 and lower, lift coefficients per wing are also enhanced during wing rotation when compared to the case of one wing rotating with the same motion. Remarkably, lift coefficients following two-winged fling during the translational phase are also enhanced when compared to the one-winged case. Indeed, they begin about 70% higher than the one-winged case during pure translation. When averaged over the entire translational part of the stroke, lift coefficients per wing are 35% higher for the two-winged case during a 4.5 chord translation following fling. In addition, lift enhancement increases with decreasing Re. This result suggests that the Weis-Fogh mechanism of lift generation has greater benefit to insects flying at lower Re. Drag coefficients produced during fling are also substantially higher for the two-winged case than the one-winged case, particularly at lower Re.


Hippocampal volumes differ across the mouse estrous cycle, can change within 24 hours, and associate with cognitive strategies.
Recent human and rodent brain imaging studies have shown that the shape of the brain can be changed by experience. These mesoscopic alterations in neuroanatomy are hypothesized to be driven by changes at the level of neuronal processes. To examine whether the shape of the brain changes rapidly, we used MRI to examine changes in the volume of the hippocampus across the 4-6 day estrous cycle in the female mouse. It is well known that changing steroid levels across the cycle influence dendritic spine maturation and alter synapse density in the hippocampus; our results show that the estrous cycle is associated with approximately 2-3% changes in hippocampal volume as seen by high-resolution ex-vivo MRI. Changes in hippocampal volume are, moreover, associated with a switch between hippocampal and striatal based navigation strategies in solving the dual choice T-maze in the same mice. A second experiment, using in-vivo MRI, suggests that these changes in hippocampal volume can occur over a 24 hour period. In summary, we show that the brain is highly plastic at a mesoscopic level of resolution detectable by MRI, that volumetric increases and decreases in hippocampal volume follow previously established patterns of changes in neuropil, and that these changes in volume predict changes in cognition.


HERG-Like potassium current regulates the resting membrane potential in glomus cells of the rabbit carotid body.
Direct evidence for a specific K(+) channel underlying the resting membrane potential in glomus cells of the carotid body has been absent. The product of the human ether-a-go-go-related gene (HERG) produces inward rectifier currents that are known to contribute to the resting membrane potential in other neuronal cells. The goal of the present study was to determine whether carotid body glomus cells express HERG-like K(+) current, and if so, to determine whether a HERG-like current regulates the resting membrane potential. Freshly dissociated rabbit glomus cells under whole cell voltage clamp exhibited slowly decaying outward currents that activated 20-30 mV positive to the resting membrane potential. Raising extracellular K(+) revealed a slowly deactivating inward tail current indicative of HERG-like K(+) current. HERG-like currents were not found in cells resembling type II cells. The HERG-like current was blocked by dofetilide (DOF) in a concentration-dependent manner (IC(50) = 13 +/- 4 nM, mean +/- SE) and high concentrations of Ba(2+) (1 and 10 mM). The biophysical and pharmacological characteristics of this inward tail current suggest that it is conducted by a HERG-like channel. The steady-state activation properties of the HERG-like current (V(h) = -44 +/- 2 mV) suggest that it is active at the resting membrane potential in glomus cells. In whole cell, current-clamped glomus cells (average resting membrane potential, - 48 +/- 4 mV), DOF, but not tetraethylammonium, caused a significant (13 mV) depolarizing shift in the resting membrane potential. Using fluorescence imaging, DOF increased [Ca(2+)](i) in isolated glomus cells. In an in-vitro carotid body preparation, DOF increased basal sensory discharge in the carotid sinus nerve in a concentration-dependent manner. These results demonstrate that glomus cells express a HERG-like current that is active at, and responsible for controlling the resting membrane potential.


Nidicolous ticks of small mammals in Anaplasma phagocytophilum-enzootic sites in northern California.
Ixodes spp. tick-borne zoonotic diseases are present across the Holarctic in humans, domestic animals, and wildlife. Small mammals are reservoirs for the rickettsial pathogen Anaplasma phagocytophilum and tick vectors may include catholic-feeding bridge vectors as well as host-specialist or nidicolous ticks. Far western North American communities in which A. phagocytophilum is maintained are complex ecologically, with multiple reservoir host and tick species, multiple strains of the bacterial pathogen A. phagocytophilum and differences in dynamics of hosts and vectors across heterogeneous landscapes. We evaluated sites in northern California in order to identify primarily nidicolous ticks and the hosts they infest. A total of 667 ticks was found in 11 study sites, including 288 on flags and 379 attached to small mammals. Larvae were over-represented among attached ticks and adults on flags. The most abundant species was I. pacificus. Two-hundred fourteen nidicolous ticks were found, most abundantly I. angustus and I. spinipalpis. All adult I. ochotonae, I. auritulus, I. angustus, I. jellisoni, and I. woodi were female, while the male:female ratio of I. spinipalpis was 1.2:1 and 1:1 for I. pacificus. The greatest number of ticks was obtained from Tamias ochrogenys, Peromyscus spp., and Neotoma fuscipes. Of 234 small mammal individuals that were infested with Ixodes spp., only 81 (34.6%) were infested with I. pacificus. The remaining infested small mammals hosted nidicolous tick species. Eight ticks were PCR-positive, including 6 I. pacificus (one adult, one larva, and 6 nymphs), and 2 adult I. ochotonae and high PCR prevalences of 18% and 9% were detected in woodrats and chipmunks, respectively. Nymphal I. angustus ticks were active year-long with a possible increase in August while larval activity was only observed in December and spring months and adults only during spring and fall. Overall, we show high tick species richness and year-round high levels of infestation on rodents by several different nidicolous ticks in areas where A. phagocytophilum is enzootic, including on reported reservoir species.


Detection and characterization of nanoparticles in suspension at low concentrations using the X-ray total scattering pair distribution function technique.
Difference atomic pair distribution function methods have been applied to detect and characterize nanoparticles suspended in a solvent at very dilute concentrations. We specifically consider nanoparticles of a pharmaceutical compound in aqueous solution using X-ray PDF methods, a challenging case due to the low atomic number of the nanoparticle species. The nanoparticles were unambiguously detected at the level of 0.25 wt%. Even at these low concentrations the signals were highly reproducible, allowing for reliable detection and quantitative analysis of the nanoparticle structure.


Effect of a Mobile Phone Intervention on Quitting Smoking in a Young Adult Population of Smokers: Randomized Controlled Trial.
Digital mobile technology presents a promising medium for reaching young adults with smoking cessation interventions because they are the heaviest users of this technology. The aim of this study was to determine the efficacy of an evidence-informed smartphone app for smoking cessation, Crush the Crave (CTC), on reducing smoking prevalence among young adult smokers in comparison with an evidence-informed self-help guide, On the Road to Quitting (OnRQ). A parallel, double-blind, randomized controlled trial with 2 arms was conducted in Canada to evaluate CTC. In total, 1599 young adult smokers (aged 19 to 29 years) intending to quit smoking in the next 30 days were recruited online and randomized to receive CTC or the control condition OnRQ for a period of 6 months. The primary outcome measure was self-reported continuous abstinence at the 6-month follow-up. Overall follow-up rates were 57.41% (918/1599) and 60.48% (967/1599) at 3 and 6 months, respectively. Moreover, 45.34% (725/1599) of participants completed baseline, 3-, and 6-month follow-up. Intention-to-treat analysis (last observation carried forward) showed that continuous abstinence (N=1599) at 6 months was not significantly different at 7.8% (64/820) for CTC versus 9.2% (72/779) for OnRQ (odds ratio; OR 0.83, 95% CI 0.59-1.18). Similarly, 30-day point prevalence abstinence at 6 months was not significantly different at 14.4% (118/820) and 16.9% (132/779) for CTC and OnRQ, respectively (OR 0.82, 95% CI 0.63-1.08). However, these rates of abstinence were favorable compared with unassisted 30-day quit rates of 11.5% among young adults. Secondary measures of quit attempts and the number of cigarettes smoked per day at 6-month follow-up did not reveal any significant differences between groups. For those who completed the 6-month follow-up, 85.1% (359/422) of young adult smokers downloaded CTC as compared with 81.8% (346/423) of OnRQ, χ21(N=845)=1.6, P=.23. Furthermore, OnRQ participants reported significantly higher levels of overall satisfaction (mean 3.3 [SD 1.1] vs mean 2.6 [SD 1.3]; t644=6.87, P<.001), perceived helpfulness (mean 5.8 [SD 2.4] vs mean 4.3 [SD 2.6], t657=8.0, P<.001), and frequency of use (mean 3.6 [SD 1.2] vs mean 3.2 [SD 1.1], t683=5.7, P<.001) compared with CTC participants. CTC was feasible for delivering cessation support but was not superior to a self-help guide in helping motivated young adults to quit smoking. CTC will benefit from further formative research to address satisfaction and usage. As smartphone apps may not serve as useful alternatives to printed self-help guides, there is a need to conduct further research to understand how digital mobile technology smoking cessation interventions for smoking cessation can be improved. ClinicalTrials.gov NCT01983150; http://clinicaltrials.gov/ct2/show/NCT01983150 (Archived by WebCite at http://www.webcitation.org/6VGyc0W0i).


Decreased bone speed of sound in children with growing pains measured by quantitative ultrasound.
The most common cause of recurring childhood musculoskeletal pain is termed growing pains (GP). We hypothesized that GP may represent a local overuse syndrome and therefore may be associated with decreased bone speed of sound (SOS) measured by quantitative ultrasound (US). We studied 39 children with GP. Bone SOS was measured by US in both mid-tibial and radius bones according to a validated protocol. Unpaired Student t test was used to compare patients and norms of healthy controls. Tibial SOS was significantly reduced in children with GP compared to controls (Z score -0.546 for boys and -0.891 for girls; p = 0.004, p < 0.001, respectively). Radius SOS was significantly reduced only in girls with GP (Z score -0.692, p = 0.006). No correlation was found between bone SOS and various demographic and clinical factors besides the child's ethnicity and body mass index. Bone SOS was significantly reduced in children with GP, especially in painful tibial regions. GP may represent a local overuse syndrome.


Reduced Sampling Size with Nanopipette for Tapping-Mode Scanning Probe Electrospray Ionization Mass Spectrometry Imaging.
Mass spectrometry imaging (MSI) with ambient sampling and ionization can rapidly and easily capture the distribution of chemical components in a solid sample. Because the spatial resolution of MSI is limited by the size of the sampling area, reducing sampling size is an important goal for high resolution MSI. Here, we report the first use of a nanopipette for sampling and ionization by tapping-mode scanning probe electrospray ionization (t-SPESI). The spot size of the sampling area of a dye molecular film on a glass substrate was decreased to 6 μm on average by using a nanopipette. On the other hand, ionization efficiency increased with decreasing solvent flow rate. Our results indicate the compatibility between a reduced sampling area and the ionization efficiency using a nanopipette. MSI of micropatterns of ink on a glass and a polymer substrate were also demonstrated.


Introduction: Self-organization in nonequilibrium chemical systems.
The field of self-organization in nonequilibrium chemical systems comprises the study of dynamical phenomena in chemically reacting systems far from equilibrium. Systematic exploration of this area began with investigations of the temporal behavior of the Belousov-Zhabotinsky oscillating reaction, discovered accidentally in the former Soviet Union in the 1950s. The field soon advanced into chemical waves in excitable media and propagating fronts. With the systematic design of oscillating reactions in the 1980s and the discovery of Turing patterns in the 1990s, the scope of these studies expanded dramatically. The articles in this Focus Issue provide an overview of the development and current state of the field.


The archaeology, chronology and stratigraphy of Madjedbebe (Malakunanja II): A site in northern Australia with early occupation.
Published ages of >50 ka for occupation at Madjedbebe (Malakunanja II) in Australia's north have kept the site prominent in discussions about the colonisation of Sahul. The site also contains one of the largest stone artefact assemblages in Sahul for this early period. However, the stone artefacts and other important archaeological components of the site have never been described in detail, leading to persistent doubts about its stratigraphic integrity. We report on our analysis of the stone artefacts and faunal and other materials recovered during the 1989 excavations, as well as the stratigraphy and depositional history recorded by the original excavators. We demonstrate that the technology and raw materials of the early assemblage are distinctive from those in the overlying layers. Silcrete and quartzite artefacts are common in the early assemblage, which also includes edge-ground axe fragments and ground haematite. The lower flaked stone assemblage is distinctive, comprising a mix of long convergent flakes, some radial flakes with faceted platforms, and many small thin silcrete flakes that we interpret as thinning flakes. Residue and use-wear analysis indicate occasional grinding of haematite and woodworking, as well as frequent abrading of platform edges on thinning flakes. We conclude that previous claims of extensive displacement of artefacts and post-depositional disturbance may have been overstated. The stone artefacts and stratigraphic details support previous claims for human occupation 50-60 ka and show that human occupation during this time differed from later periods. We discuss the implications of these new data for understanding the first human colonisation of Sahul.


Stagnation in the decline of the world population growth rate during the 1980s.
The world human population growth rate after World War II passed through three phases: the rise in the 1950s and 1960s, the fall (though still at a positive level) in the 1970s, and the plateau in the 1980s. The rise was produced by the global decline in death rates, the fall was mainly due to the reduction of fertility in a number of developing countries, and the stagnation of growth rate decline was attributable to three major factors. First, substantial fertility declines started around 1970 and stalled around 1980 in both China and India. Second, the age structure of population changed in favor of higher birth rates. Third, although fertility started to decline significantly around 1970 mainly in East Asia, Southeast Asia, and Latin America, few countries have begun fertility declines since then. Many countries in sub-Saharan Africa and South Asia have not started substantial fertility reductions, deepening the gap between developing countries that are moving to lower fertility levels and those that are left behind.


Characterization of the influence of chlororespiration on the regulation of photosynthesis in the glaucophyte Cyanophora paradoxa.
Glaucophytes are primary symbiotic algae with unique plastids called cyanelles, whose structure is most similar to ancestral cyanobacteria among plastids in photosynthetic organisms. Here we compare the regulation of photosynthesis in glaucophyte with that in cyanobacteria in the aim of elucidating the changes caused by the symbiosis in the interaction between photosynthetic electron transfer and other metabolic pathways. Chlorophyll fluorescence measurements of the glaucophyte Cyanophora paradoxa NIES-547 indicated that plastoquinone (PQ) pool in photosynthetic electron transfer was reduced in the dark by chlororespiration. The levels of nonphotochemical quenching of chlorophyll fluorescence was high in the dark but decreased under low light, and increased again under high light. This type of concave light dependence was quite similar to that observed in cyanobacteria. Moreover, the addition of ionophore hardly affected nonphotochemical quenching, suggesting state transition as a main component of the regulatory system in C. paradoxa. These results suggest that cyanelles of C. paradoxa retain many of the characteristics observed in their ancestral cyanobacteria. From the viewpoint of metabolic interactions, C. paradoxa is the primary symbiotic algae most similar to cyanobacteria than other lineages of photosynthetic organisms.


Activation of response force by self-splitting objects: where are the limits of feedforward Gestalt processing?
Most objects can be recognized easily even when they are partly occluded. This also holds when several overlapping objects share the same surface features (self-splitting objects) which is an illustration of the grouping principle of Good Gestalt. We employed outline and filled contour stimuli in a primed flanker task to test whether the processing of self-splitting objects is in accordance with a simple feedforward model. We obtained priming effects in response time and response force for both types of stimuli, even when increasing the number of occluders up to three. The results for outline contours were in full accordance with a feedforward account. This was not the case for the results for filled contours (i.e., for self-splitting objects), especially under conditions of strong occlusion. We conclude that the implementation of the Good Gestalt principle is fast but still based on recurrent processing.


Effect of 2 Enteral Feeding Schedules on Intra-abdominal Pressure in Patients Receiving Mechanical Ventilation: A Randomized Controlled Trial.
Elevated intra-abdominal pressure is associated with morbidity in critically ill patients. Enteral feeding is important for these patients but may cause complications. To compare the effects of 2 intermittent feeding schedules on intra-abdominal pressure in patients receiving mechanical ventilation. A single-blinded, prospective, parallel-group, randomized controlled trial was conducted in an intensive care unit in a teaching hospital in Egypt. Fifty adult patients requiring more than 48 hours of mechanical ventilation were enrolled. Patients were randomly sorted into 2 study groups. The intervention group received intermittent enteral feedings 5 times daily at 4-hour intervals. The control group received intermittent enteral feedings 10 times daily at 2-hour intervals. Both groups fasted for 8 hours overnight. Intra-abdominal pressure was measured 7 times: at admission and before and after the first 3 feedings on the third day of the intensive care unit stay. One-way repeated-measures analysis of variance showed that mean intra-abdominal pressure was higher in the control group before the first feeding (t = 2.27, P = .03) but was higher in the intervention group after the second feeding (t = 2.51, P = .02) and after the third feeding (t = 2.41, P = .02). Vomiting and diarrhea were not significantly different between the groups. More constipation and abdominal distension occurred in the intervention group than in the control group. Intra-abdominal pressure was not significantly affected by feeding interval. However, reducing the time interval between intermittent enteral feedings may minimize the risk for constipation and abdominal distension.


Impact of prefrontal cortex in nicotine-induced excitation of ventral tegmental area dopamine neurons in anesthetized rats.
Systemic administration of nicotine increases dopaminergic (DA) neuron firing in the ventral tegmental area (VTA), which is thought to underlie nicotine reward. Here, we report that the medial prefrontal cortex (mPFC) plays a critical role in nicotine-induced excitation of VTA DA neurons. In chloral hydrate-anesthetized rats, extracellular single-unit recordings showed that VTA DA neurons exhibited two types of firing responses to systemic nicotine. After nicotine injection, the neurons with type-I response showed a biphasic early inhibition and later excitation, whereas the neurons with type-II response showed a monophasic excitation. The neurons with type-I, but not type-II, response exhibited pronounced slow oscillations (SOs) in firing. Pharmacological or structural mPFC inactivation abolished SOs and prevented systemic nicotine-induced excitation in the neurons with type-I, but not type-II, response, suggesting that these VTA DA neurons are functionally coupled to the mPFC and nicotine increases firing rate in these neurons in part through the mPFC. Systemic nicotine also increased the firing rate and SOs in mPFC pyramidal neurons. mPFC infusion of a non-α7 nicotinic acetylcholine receptor (nAChR) antagonist mecamylamine blocked the excitatory effect of systemic nicotine on the VTA DA neurons with type-I response, but mPFC infusion of nicotine failed to excite these neurons. These results suggest that nAChR activation in the mPFC is necessary, but not sufficient, for systemic nicotine-induced excitation of VTA neurons. Finally, systemic injection of bicuculline prevented nicotine-induced firing alterations in the neurons with type-I response. We propose that the mPFC plays a critical role in systemic nicotine-induced excitation of VTA DA neurons.


Advances in the biological therapy and gene therapy of malignant disease.
Biological and gene therapy of cancer have become important components of clinical cancer research. Advances in this area are based on evidence for the presence of tumor antigens, antitumor immune responses, evasion of host control by tumors, and the recognition of host defense failure in cancer patients. These mechanisms are being corrected or exploited in the development of biological and gene therapy. Over the last decade, 9 biological therapies have received Food and Drug Administration approval, and another 12 appear promising and will likely be approved in the next few years. Our approach to gene therapy has been to allogenize tumors by the direct intratumoral injection of HLA-B7/beta2-microglobulin genes as plasmid DNA in a cationic lipid into patients with malignant melanoma. In four Phase I studies, we found a 36% response by the local injected tumor and a 19% systemic antitumor response. In other cancers, gene transfer, expression, and an intratumoral T-cell response were seen, but no clinical response was seen. A variety of follow-up studies with HLA-B7 and other genes are planned. Evasion of host control is now a major target of gene therapy. Strategies to overcome this include up-regulation of MHC and introduction of cell adhesion molecules into tumor cells, suppression of transforming growth factor and interleukin 10 production by tumor cells, and blockade of the fas ligand-fas interaction between tumor cells and attacking lymphocytes. With these approaches, it seems likely that gene therapy may become the fifth major modality of cancer treatment in the next decade.


Long-term safety and efficacy of drug-eluting versus bare-metal stents in Sweden.
The long-term safety and efficacy of drug-eluting coronary stents have been questioned. We evaluated 47,967 patients in Sweden who received a coronary stent and were entered into the Swedish Coronary Angiography and Angioplasty Registry between 2003 and 2006 and for whom complete follow-up data were available for 1 to 5 years (mean, 2.7). In the primary analysis, we compared patients who received one drug-eluting coronary stent (10,294 patients) with those who received one bare-metal stent (18,659), after adjustment for differences in clinical characteristics of the patients and characteristics of the vessels and lesions. Analyses of outcome were based on 2380 deaths and 3198 myocardial infarctions. There was no overall difference between the group that received drug-eluting stents and the group that received bare-metal stents in the combined end point of death or myocardial infarction (relative risk with drug-eluting stents, 0.96; 95% confidence interval [CI], 0.89 to 1.03) or the individual end points of death (relative risk, 0.94; 95% CI, 0.85 to 1.05) and myocardial infarction (relative risk, 0.97; 95% CI, 0.88 to 1.06), and there was no significant difference in outcome among subgroups stratified according to the indication for stent implantation. Patients who received drug-eluting stents in 2003 had a significantly higher rate of late events than patients who received bare-metal stents in the same year, but we did not observe any difference in outcome among patients treated in later years. The average rate of restenosis during the first year was 3.0 events per 100 patient-years with drug-eluting stents versus 4.7 with bare-metal stents (adjusted relative risk, 0.43; 95% CI, 0.36 to 0.52); 39 patients would need to be treated with drug-eluting stents to prevent one case of restenosis. Among high-risk patients, the adjusted risk of restenosis was 74% lower with drug-eluting stents than with bare-metal stents, and only 10 lesions would need to be treated to prevent one case of restenosis. As compared with bare-metal stents, drug-eluting stents are associated with a similar long-term incidence of death or myocardial infarction and provide a clinically important decrease in the rate of restenosis among high-risk patients.


Inactivation of the inhA-encoded fatty acid synthase II (FASII) enoyl-acyl carrier protein reductase induces accumulation of the FASI end products and cell lysis of Mycobacterium smegmatis.
The mechanism of action of isoniazid (INH), a first-line antituberculosis drug, is complex, as mutations in at least five different genes (katG, inhA, ahpC, kasA, and ndh) have been found to correlate with isoniazid resistance. Despite this complexity, a preponderance of evidence implicates inhA, which codes for an enoyl-acyl carrier protein reductase of the fatty acid synthase II (FASII), as the primary target of INH. However, INH treatment of Mycobacterium tuberculosis causes the accumulation of hexacosanoic acid (C(26:0)), a result unexpected for the blocking of an enoyl-reductase. To test whether inactivation of InhA is identical to INH treatment of mycobacteria, we isolated a temperature-sensitive mutation in the inhA gene of Mycobacterium smegmatis that rendered InhA inactive at 42 degrees C. Thermal inactivation of InhA in M. smegmatis resulted in the inhibition of mycolic acid biosynthesis, a decrease in hexadecanoic acid (C(16:0)) and a concomitant increase of tetracosanoic acid (C(24:0)) in a manner equivalent to that seen in INH-treated cells. Similarly, INH treatment of Mycobacterium bovis BCG caused an inhibition of mycolic acid biosynthesis, a decrease in C(16:0), and a concomitant accumulation of C(26:0). Moreover, the InhA-inactivated cells, like INH-treated cells, underwent a drastic morphological change, leading to cell lysis. These data show that InhA inactivation, alone, is sufficient to induce the accumulation of saturated fatty acids, cell wall alterations, and cell lysis and are consistent with InhA being a primary target of INH.


Risk-stratification methods for identifying patients for care coordination.
Care coordination is a key component of the patient-centered medical home. However, the mechanism for identifying primary care patients who may benefit the most from this model of care is unclear. To evaluate the performance of several risk-adjustment/stratification instruments in predicting healthcare utilization. Retrospective cohort analysis. All adults empaneled in 2009 and 2010 (n = 83,187) in a primary care practice were studied. We evaluated 6 models: Adjusted Clinical Groups (ACGs), Hierarchical Condition Categories (HCCs), Elder Risk Assessment, Chronic Comorbidity Count, Charlson Comorbidity Index, and Minnesota Health Care Home Tiering. A seventh model combining Minnesota Tiering with ERA score was also assessed. Logistic regression models using demographic characteristics and diagnoses from 2009 were used to predict healthcare utilization and costs for 2010 with binary outcomes (emergency department [ED] visits, hospitalizations, 30-day readmissions, and highcost users in the top 10%), using the C statistic and goodness of fit among the top decile. The ACG model outperformed the others in predicting hospitalizations with a C statistic range of 0.67 (CMS-HCC) to 0.73. In predicting ED visits, the C statistic ranged from 0.58 (CMSHCC) to 0.67 (ACG). When predicting the top 10% highest cost users, the performance of the ACG model was good (area under the curve = 0.81) and superior to the others. Although ACG models generally performed better in predicting utilization, use of any of these models will help practices implement care coordination more efficiently.


Automatic parameter selection for multimodal image registration.
Over the past ten years similarity measures based on intensity distributions have become state-of-the-art in automatic multimodal image registration. An implementation for clinical usage has to support a plurality of images. However, a generally applicable parameter configuration for the number and sizes of histogram bins, optimal Parzen-window kernel widths or background thresholds cannot be found. This explains why various research groups present partly contradictory empirical proposals for these parameters. This paper proposes a set of data-driven estimation schemes for a parameter-free implementation that eliminates major caveats of heuristic trial and error. We present the following novel approaches: a new coincidence weighting scheme to reduce the influence of background noise on the similarity measure in combination with Max-Lloyd requantization, and a tradeoff for the automatic estimation of the number of histogram bins. These methods have been integrated into a state-of-the-art rigid registration that is based on normalized mutual information and applied to CT-MR, PET-MR, and MR-MR image pairs of the RIRE 2.0 database. We compare combinations of the proposed techniques to a standard implementation using default parameters, which can be found in the literature, and to a manual registration by a medical expert. Additionally, we analyze the effects of various histogram sizes, sampling rates, and error thresholds for the number of histogram bins. The comparison of the parameter selection techniques yields 25 approaches in total, with 114 registrations each. The number of bins has no significant influence on the proposed implementation that performs better than both the manual and the standard method in terms of acceptance rates and target registration error (TRE). The overall mean TRE is 2.34 mm compared to 2.54 mm for the manual registration and 6.48 mm for a standard implementation. Our results show a significant TRE reduction for distortion-corrected magnetic resonance images.


Hepatic metabolism of aminopyrine in patients with chronic renal failure.
1. To evaluate potential alterations in hepatic metabolism of drugs occurring in patients with renal insufficiency the fate of aminopyrine was studied in 17 patients with chronic renal failure and in 27 normal subjects. 2. Although patients with chronic renal failure exhibited large variations, their aminopyrine plasma disappearance times (mean 0.62 +/- SD 0.24 h-1) were significantly higher than those found in normal subjects (0.30 +/- 0.07 h-1, P less than 0.002). 3. 14CO2 derived from [dimethylamine-14C]aminopyrine disappeared from breath more rapidly in patients with chronic renal failure and a history of analgesic abuse (0.40 +/- 0.04 h-1) than in control subjects (0.22 +/- 0.03 h-1, P less than 0.01) and in other patients with chronic renal failure (0.24 +/- 0.04 h-1). 4. Dialysis treatment and serum creatinine concentrations were not correlated with the rates of aminopyrine metabolism. Two additional patients, however, with combined renal and hepatic disease, exhibited markedly slowed rates of aminopyrine demethylation. 5. Although chronic renal failure by itself might not alter microsomal drug metabolism it is concluded that, in patients with a history of abuse of phenacetin-containing analgesics, marked acceleration in aminopyrine N-demethylation may be observed.


IS THERE EVIDENCE FOR AN ASSOCIATION BETWEEN CHANGES IN TRAINING LOAD AND RUNNING-RELATED INJURIES? A SYSTEMATIC REVIEW.
Sudden changes (increases and decreases) in training load have been suggested to play a key role in the development of running-related injuries. However, the compiled evidence for an association between change in training load and running-related injury does not exist. The purpose of the present systematic review was to compile the evidence from original articles examining the association between changes in training load and running-related injuries. Systematic review. Four databases (Pubmed/Medline, SPORTDiscus, Embase, and Scopus) were systematically searched. Two reviewers screened titles, abstracts, and full-text articles independently. Articles were included if i) the study design was a randomized trial, a prospective cohort study, a cross-sectional study or a case-control study, ii) participants were runners between 18-65 years, and iii) specific information on changes in training load was provided. Methodological quality of included articles was assessed using the Newcastle Ottawa Scale and the PEDro rating scale. Four articles fulfilled the eligibility criteria of which three found an association between increases in training load and an increased risk of running-related injuries: This association was shown by an increased injury risk amongst runners: i) if they recently had performed one or more changes in either velocity and/or distance and/or frequency compared with the non-injured runners (p = 0.037), ii) increasing their average weekly running distance by more than 30% compared to an increase less than 10% (Hazard Ratio = 1.59 (95% Confidence Interval: 0.96; 2.66)), iii) increasing their total running distance significantly more the week before the injury origin compared with other weeks (mean difference: 86%; 95% Confidence Interval: 12%; 159%, p = 0.026). However, no difference was found between a 10% and a 24% average increase in weekly volume (HR = 0.8, 95% CI: 0.6; 1.3). Very limited evidence exists supporting that a sudden change in training load is associated with increased risk of running-related injury. 2.


Native LDL promotes differentiation of human monocytes to macrophages with an inflammatory phenotype.
Recruitment of monocytes in atherosclerosis is dependent upon increased levels of plasma lipoproteins which accumulate in the blood vessel wall. The extracellular milieu can influence the phenotype of monocyte subsets (classical: CD14++CD16-, intermediate: CD14+CD16+ and non-classical: CD14dimCD16++) and macrophages (M1 or M2) and consequently the initiation, progression and/or regression of atherosclerosis. However, it is not known what effect lipoproteins, in particular native low-density lipoproteins (nLDL), have on the polarisation of monocyte-derived macrophages. Monocytes were differentiated into macrophages in the presence of nLDL. nLDL increased gene expression of the inflammatory cytokines TNFα and IL-6 in macrophages polarised towards the M1 phenotype while decreasing the M2 surface markers, CD206 and CD200R and the anti-inflammatory cytokines TGFβ and IL-10. Compared to the classical and intermediate subsets, the non-classical subset-derived macrophages had a reduced ability to respond to M1 stimuli (LPS and IFNγ). nLDL enhanced the TNFα and IL-6 gene expression in macrophages from all monocyte subsets, indicating an inflammatory effect of nLDL. Further, the classical and intermediate subsets both responded to M2 stimuli (IL-4) with upregulation of TGFβ and SR-B1 mRNA; an effect, which was reduced by nLDL. In contrast, the non-classical subset failed to respond to IL-4 or nLDL, suggesting it may be unable to polarise into M2 macrophages. Our data suggests that monocyte interaction with nLDL significantly affects macrophage polarisation and that this interaction appears to be subset dependent.


Coronary artery disease in severe haemophilia.
It has been suggested that the coagulation defect in haemophilia A protects these patients from coronary artery disease. We report two patients with severe haemophilia, who each had severe atheroma localised to the coronary arteries. Both patients had received only small amounts of factor VIII concentrates in their lifetime because of various social reasons. In one case death resulted from myocardial infarction secondary to an occlusive thrombus in a coronary artery. The implications of these findings to the pathogenesis of vascular disease are discussed.


Cyclist-motorist crash patterns in Denmark: a latent class clustering approach.
The current study aimed at uncovering patterns of cyclist-motorist crashes in Denmark and investigating their prevalence and severity. The importance of implementing clustering techniques for providing a holistic overview of vulnerable road users' crash patterns derives from the need to prioritize safety issues and to devise efficient preventive measures. The current study focused on cyclist-motorist crashes that occurred in Denmark during the period between 2007 and 2011. To uncover crash patterns, the current analysis applied latent class clustering, an unsupervised probabilistic clustering approach that relies on the statistical concept of likelihood and allows partial overlap across clusters. The analysis yielded 13 distinguishable cyclist-motorist latent classes. Specific crash patterns for urban and rural areas were revealed. Prevalent features that allowed differentiating the latent classes were speed limit, infrastructure type, road surface conditions, number of lanes, motorized vehicle precrash maneuvers, the availability of a cycle lane, cyclist intoxication, and helmet wearing behavior. After the latent class clustering, the distribution of cyclists' injury severity within each cluster was analyzed. The latent class clustering approach provided a comprehensive and clear map of cyclist-motorist crash patterns. The results are useful for prioritizing and resolving safety issues in urban areas, where there is a significant share of cyclists potentially involved in multiple hazardous situations or where extensive bicycle sharing programs are planned.


Bdellovibrio bacteriovorus HD100 guards against Pseudomonas tolaasii brown-blotch lesions on the surface of post-harvest Agaricus bisporus supermarket mushrooms.
Pseudomonas tolaasii is a problematic pathogen of cultured mushrooms, forming dark brown 'blotches' on mushroom surfaces and causing spoilage during crop growth and post-harvest . Treating P. tolaasii infection is difficult, as other, commensal bacterial species such as Pseudomonas putida are necessary for mushroom growth, so treatments must be relatively specific. We have found that P. tolaasii is susceptible to predation in vitro by the δ-proteobacterium Bdellovibrio bacteriovorus. This effect also occurred in funga, where B. bacteriovorus was administered to post-harvest mushroom caps before and after administration of the P. tolaasii pathogen. A significant, visible improvement in blotch appearance, after incubation, was observed on administration of Bdellovibrio. A significant reduction in viable P. tolaasii cell numbers, recovered from the mushroom tissue, was detected. This was accompanied by a more marked reduction in blotch severity on Bdellovibrio administration. We found that there was in some cases an accompanying overgrowth of presumed-commensal, non-Pseudomonas bacteria on post-harvest mushroom caps after Bdellovibrio-treatment. These bacteria were identified (by 16SrRNA gene sequencing) as Enterobacter species, which were seemingly resistant to predation. We visualised predatory interactions occuring between B. bacteriovorus and P. tolaasii on the post-harvest mushroom cap surface by Scanning Electron Microscopy, seeing predatory invasion of P. tolaasii by B. bacteriovorus in funga. This anti-P. tolaasii effect worked well in post-harvest supermarket mushrooms, thus Bdellovibrio was not affected by any pre-treatment of mushrooms for commercial/consumer purposes. The soil-dwelling B. bacteriovorus HD100 preys upon and kills P. tolaasii, on mushroom surfaces, and could therefore be applied to prevent spoilage in post-harvest situations where mushrooms are stored and packaged for sale.


Regulation of rat liver apolipoprotein A-I, apolipoprotein A-II and acyl-coenzyme A oxidase gene expression by fibrates and dietary fatty acids.
The regulation by fibrates and dietary fatty acids of the hepatic gene expression of apolipoproteins (apo) A-I and A-II, the major protein constituents of high-density lipoproteins, as well as of acyl-CoA oxidase, the rate-limiting enzyme of the peroxisomal beta-oxidation pathway, was studied in vivo in the rat and in vitro in primary cultures of rat hepatocytes. In primary hepatocytes, different fibrates decreased apo A-I and increased acyl-CoA oxidase mRNA levels, whereas apo A-II mRNA only decreased in level after treatment with fenofibric acid, but not after bezafibrate, gemfibrozil or Wy-14643 treatment. Treatment with fenofibric acid counteracted the increase in apo A-I mRNA levels observed after dexamethasone or all-trans retinoic acid treatment, whereas simultaneous addition of fenofibric acid together with all-trans retinoic acid or dexamethasone resulted in a superinduction of acyl-CoA oxidase mRNA. Addition of the n-3 polyunsaturated fatty acids (PUFAs), docosanohexaenoic acid and eicosanopentaenoic acid, or the fatty acid derivative alpha-bromopalmitate, decreased apo A-I and increased acyl-CoA oxidase mRNA in a dose-dependent and time-dependent manner, whereas apo A-II mRNA did not change significantly. Nuclear run-on experiments demonstrated that fenofibric acid and alpha-bromopalmitate decreased apo A-I and increased acyl-CoA oxidase gene expression at the transcriptional level. When rats were fed isocaloric diets enriched in saturated fat (hydrogenated coconut oil), n-6 PUFAs (safflower oil) or n-3 PUFAs (fish oil), a significant decrease in liver apo A-I and apo A-II mRNA levels was only observed after fish oil feeding. Compared to feeding low fat, liver acyl-CoA oxidase mRNA increased after fat feeding, but this effect was most pronounced (twofold) in rats fed fish oil. Results from these studies indicate that fish oil feeding reduces rat liver apo A-I and apo A-II gene expression, similar to results obtained after feeding fenofibrate. Fibrates and n-3 fatty acids (and the fatty acid derivative, alpha-bromopalmitate) down-regulate apo A-I and induce acyl-CoA oxidase gene expression through a direct transcriptional action on the hepatocyte. In contrast, only fenofibric acid, but not the other fibrates or fatty acids tested, decrease apo A-II gene expression in vitro.


Mechanism of free fatty acid-induced insulin resistance in humans.
To examine the mechanism by which lipids cause insulin resistance in humans, skeletal muscle glycogen and glucose-6-phosphate concentrations were measured every 15 min by simultaneous 13C and 31P nuclear magnetic resonance spectroscopy in nine healthy subjects in the presence of low (0.18 +/- 0.02 mM [mean +/- SEM]; control) or high (1.93 +/- 0.04 mM; lipid infusion) plasma free fatty acid levels under euglycemic (approximately 5.2 mM) hyperinsulinemic (approximately 400 pM) clamp conditions for 6 h. During the initial 3.5 h of the clamp the rate of whole-body glucose uptake was not affected by lipid infusion, but it then decreased continuously to be approximately 46% of control values after 6 h (P < 0.00001). Augmented lipid oxidation was accompanied by a approximately 40% reduction of oxidative glucose metabolism starting during the third hour of lipid infusion (P < 0.05). Rates of muscle glycogen synthesis were similar during the first 3 h of lipid and control infusion, but thereafter decreased to approximately 50% of control values (4.0 +/- 1.0 vs. 9.3 +/- 1.6 mumol/[kg.min], P < 0.05). Reduction of muscle glycogen synthesis by elevated plasma free fatty acids was preceded by a fall of muscle glucose-6-phosphate concentrations starting at approximately 1.5 h (195 +/- 25 vs. control: 237 +/- 26 mM; P < 0.01). Therefore in contrast to the originally postulated mechanism in which free fatty acids were thought to inhibit insulin-stimulated glucose uptake in muscle through initial inhibition of pyruvate dehydrogenase these results demonstrate that free fatty acids induce insulin resistance in humans by initial inhibition of glucose transport/phosphorylation which is then followed by an approximately 50% reduction in both the rate of muscle glycogen synthesis and glucose oxidation.


Pharmacovigilance: a worldwide master key for drug safety monitoring.
Pharmacovigilance is like a sunshade to describe the processes for monitoring and evaluating ADRs and it is a key component of effective drug regulation systems, clinical practice and public health programmes. The number of Adverse Drug Reactions (ADRs) reported resulted in an increase in the volume of data handled, and to understand the pharmacovigilance, a high level of expertise is required to rapidly detect drug risks as well as to defend the product against an inappropriate removal. The current global network of pharmacovigilance centers, coordinated by the Uppsala Monitoring Centre, would be strengthened by an independent system of review. This would consider litigious and important drug safety issues that have the potential to affect public health adversely beyond national boundaries. Recently, pharmacovigilance has been confined, mainly to detect adverse drug events that were previously either unknown or poorly understood. Pharmacovigilance is an important and integral part of clinical research and these days it is growing in many countries. Today many pharmacovigilance centers are working for drug safety monitoring in this global pitch, however, at the turn of the millennium pharmacovigilance faces major challenges in aspect of better safety and monitoring of drugs. In this review we will discuss about drug safety, worldwide pharmacovigilance centers and their role, benefits and challenges of pharmacovigilance and its future consideration in healthcare sectors.


When ageing and disasters collide: lessons from 16 international case studies.
Sixteen case studies examined the impact of various natural disasters and conflict-related emergencies on older people, the strengths and gaps in emergency planning, response and recovery, and the contributions older people made to their families and communities. Case examples were chosen from both developed and developing countries. Older persons suffered disproportionate impacts in several cases. Regardless of the country's level of prosperity, those most affected tended to be economically disadvantaged, disabled or frail, women, socially isolated, or caregivers of family members. Emergency responders were often not aware of distinct needs or abilities of older persons and not equipped to respond appropriately. The best emergency practices recognised and included specific needs within mainstream efforts and integrated older persons in community planning, response and recovery activities. This paper presents the 'lessons learned' from these case studies and makes the case for greater attention to this segment of the population in emergency management.


Solutions for filling gaps in accountable care measure sets.
A primary objective of accountable care is to support providers in reforming care to improve outcomes and lower costs. Gaps in accountable care measure sets may cause missed opportunities for improvement and missed signals of problems in care. Measures to balance financial incentives may be particularly important for high-cost conditions or specialty treatments. This study explored gaps in measure sets for specific conditions and offers strategies for more comprehensive measurement that do not necessarily require more measures. A descriptive analysis of measure gaps in accountable care programs and proposed solutions for filling the gaps. We analyzed gaps in 2 accountable care organization measure sets for 20 high-priority clinical conditions by comparing the measures in those sets with clinical guidelines and assessing the use of outcome measures. Where we identified gaps, we looked for existing measures to address the gaps. Gaps not addressed by existing measures were considered areas for measure development or measurement strategy refinement. We found measure gaps across all 20 conditions, including those conditions that are commonly addressed in current measure sets. In addition, we found many gaps that could not be filled by existing measures. Results across all 20 conditions informed recommendations for measure set improvement. Addressing all gaps in accountable care measure sets with more of the same types of measures and approaches to measurement would require an impractical number of measures and would miss the opportunity to use better measures and innovative approaches. Strategies for effectively filling measure gaps include using preferred measure types such as cross-cutting, outcome, and patient-reported measures. Program implementers should also apply new approaches to measurement, including layered and modular models.


Muscarinic excitation-contraction coupling mechanisms in tracheal and bronchial smooth muscles.
We investigated the mechanisms underlying muscarinic excitation-contraction coupling in canine airway smooth muscle using organ bath, fura 2 fluorimetric, and patch-clamp techniques. Cyclopiazonic acid (CPA) augmented the responses to submaximal muscarinic stimulation in both tracheal (TSM) and bronchial smooth muscles (BSM), consistent with disruption of the barrier function of the sarcoplasmic reticulum. During maximal stimulation, however, CPA evoked substantial relaxation in TSM but not BSM. CPA reversal of carbachol tone persisted in the presence of tetraethylammoium or high KCl, suggesting that hyperpolarization is not involved; CPA relaxations were absent in tissues preconstricted with KCl alone or by permeabilization with beta-escin, ruling out a nonspecific effect on the contractile apparatus. Peak contractions were sensitive to inhibitors of tyrosine kinase (genistein) or Rho kinase (Y-27632). Sustained responses were dependent on Ca(2+) influx in TSM but not BSM; this influx was sensitive to Ni(2+) but not La(3+). In conclusion, there are several mechanisms underlying excitation-contraction coupling in airway smooth muscle, the relative importance of which varies depending on tissue and degree of stimulation.


A summary of the literature on global hearing impairment: current status and priorities for action.
Hearing loss (HL) and deafness are global issues that affect at least 278 million people worldwide. Two thirds of the people who have HL worldwide live in developing countries. Importantly, it is estimated that 50% of this HL can be prevented. In developing countries, funding for prevention, early detection, and rehabilitative programs is severely limited, and therefore, agencies must compete against priorities to treat life-threatening, pandemic diseases such as human immunodeficiency virus, malaria, and tuberculosis. Delays in diagnosis are common, and social attitudes, local customs, and cultural bias are contributing factors. The purpose of this review is to gain an understanding of the prevalence of HL in the developing world and to focus attention on the growing need for both prevention and effective treatment programs. A second goal is to use this information to suggest priorities and approaches to address these problems worldwide. The data were compiled from a review of the literature on the global impacts of hearing impairment and recently published reports on the prevalence and cause of hearing impairment in developing nations. The high prevalence of HL in the developing world is due to a variety of factors, including lack of widespread comprehensive immunization programs and other medical care, and inadequate funds for intervention once HL is identified. International organizations, governments, and nongovernment organizations have many opportunities to prevent and treat HL through cost-effective means.


L-arginine is required for expression of the activated macrophage effector mechanism causing selective metabolic inhibition in target cells.
L-Arginine is required for expression of the activated macrophage cytotoxic effector mechanism that causes inhibition of mitochondrial respiration, aconitase activity, and DNA synthesis in tumor target cells. This effector mechanism is active in the presence of L-arginine even when the cocultivation medium lacks all other amino acids and serum. Cytotoxic activated macrophage-induced inhibition of mitochondrial respiration in target cells is proportional to the concentration of L-arginine in the medium. L-Arginine must be present during the cocultivation period. Pretreatment of cytotoxic activated macrophages with L-arginine or posttreatment of the target cells after cocultivation is not effective. D-Arginine does not substitute for L-arginine and at high concentrations is a competitive inhibitor of the L-arginine-dependent effector mechanism. Other analogues that could not replace L-arginine include agmatine, argininic acid, arginine hydroxamate, and tosyl-L-arginine methyl ester. L-homoarginine, however, can effectively substitute for L-arginine. NG-monomethyl-L-arginine is a potent competitive inhibitor of this effector mechanism. High concentrations of lipopolysaccharide do not reverse inhibition of the L-arginine-dependent effector mechanism by NG-monomethyl-L-arginine. However, inhibition of the effector mechanism by NG-monomethyl-L-arginine can be overridden by increasing the concentration of L-arginine in the culture medium. We compared NGNG-dimethyl-L-arginine and NGN1G-dimethyl-L-arginine with NG-monomethyl-L-arginine as inhibitors of the L-arginine-dependent effector mechanism. The results show that the inhibitory effect of these guanidino methylated derivatives of L-arginine is highly determined by structure. Guanidine is a weak competitive inhibitor of the L-arginine-dependent effector mechanism. The requirement for L-arginine does not appear to be for protein synthesis, creatine biosynthesis, polyamine biosynthesis, or ADP ribosylation reactions. Bacterial lipopolysaccharide is effective as a second signal only when the cocultivation medium contains L-arginine, and this strict L-arginine dependency is not overridden by increasing the concentration of lipopolysaccharide. Bovine liver arginase, by competing for L-arginine in the cocultivation medium, inhibits the L-arginine-dependent activated macrophage cytotoxic effector mechanism.


Comparative analysis of RNAi screening technologies at genome-scale reveals an inherent processing inefficiency of the plasmid-based shRNA hairpin.
RNAi screening in combination with the genome-sequencing projects would constitute the Holy Grail of modern genetics; enabling discovery and validation towards a better understanding of fundamental biology leading to novel targets to combat disease. Hit discordance at inter-screen level together with the lack of reproducibility is emerging as the technology's main pitfalls. To examine some of the underlining factors leading to such discrepancies, we reasoned that perhaps there is an inherent difference in knockdown efficiency of the various RNAi technologies. For this purpose, we utilized the two most popular ones, chemically synthesized siRNA duplex and plasmid-based shRNA hairpin, in order to perform a head to head comparison. Using a previously developed gain-of-function assay probing modulators of the miRNA biogenesis pathway, we first executed on a siRNA screen against the Silencer Select V4.0 library (AMB) nominating 1,273, followed by an shRNA screen against the TRC1 library (TRC1) nominating 497 gene candidates. We observed a poor overlap of only 29 hits given that there are 15,068 overlapping genes between the two libraries; with DROSHA as the only common hit out of the seven known core miRNA biogenesis genes. Distinct genes interacting with the same biogenesis regulators were observed in both screens, with a dismal cross-network overlap of only 3 genes (DROSHA, TGFBR1, and DIS3). Taken together, our study demonstrates differential knockdown activities between the two technologies, possibly due to the inefficient intracellular processing and potential cell-type specificity determinants in generating intended targeting sequences for the plasmid-based shRNA hairpins; and suggests this observed inefficiency as potential culprit in addressing the lack of reproducibility.


Defocus morphing in real aperture images.
A new concept called defocus morphing in real aperture images is introduced. View morphing is an existing example of shape-preserving image morphing based on the motion cue. It is proved that images can also be morphed based on the depth-related defocus cue. This illustrates that the morphing operation is not necessarily a geometric process alone; one can also perform a photometry-based morphing wherein the shape information is implicitly buried in the image intensity field. A theoretical understanding of the defocus morphing process is presented. It is shown mathematically that, given two observations of a three-dimensional scene for different camera parameter settings, we can obtain a virtual observation for any camera parameter setting through a simple nonlinear combination of these observations.