Sunbeds and carcinogenesis: the need for new regulations and restrictions in Europe from the Euromelanoma perspective.
Experimental investigations have definitely assessed that ultraviolet A (UVA) as well as ultraviolet B (UVB) radiation induce mutagenic DNA photoproducts and other cell damages with a carcinogenic potential. Artificial tanning increases significantly the lifetime risk for basal cell carcinoma, squamous cell carcinoma and melanoma particularly in subjects with fair skin type, subjects with a history of skin cancer or frequent childhood sunburn or if exposures took place at an age younger than 18 years. In addition, experimental and clinical evidence indicate that UVA exposure promotes skin photoageing. Therefore we are dealing with a recreational activity (for customers) and a profitable business (for the tanning industry) with human costs, i.e. an increase in morbidity and mortality by skin cancer, and health and social costs leading to an increased expenditure by the European national health systems. In a few European countries, legislation has recently prohibited the use of sunbeds for minors, pregnant women, people with skin cancer or a history of skin cancer and individuals who do not tan or who burn easily from sun exposure. However, this legislation seems to be insufficient from a photobiological perspective, and importantly, it is largely disregarded by consumers and tanning industry. Therefore the Euromelanoma group proposes a new, more stringent regulation for the tanning industry and restrictions for customers, particularly for those individuals with constitutional and anamnestic risk factors. Finally, we ask for an enhanced commitment to increase the awareness of the general population on the risk of artificial tanning.


Long noncoding RNAs: cellular address codes in development and disease.
In biology as in real estate, location is a cardinal organizational principle that dictates the accessibility and flow of informational traffic. An essential question in nuclear organization is the nature of the address code--how objects are placed and later searched for and retrieved. Long noncoding RNAs (lncRNAs) have emerged as key components of the address code, allowing protein complexes, genes, and chromosomes to be trafficked to appropriate locations and subject to proper activation and deactivation. lncRNA-based mechanisms control cell fates during development, and their dysregulation underlies some human disorders caused by chromosomal deletions and translocations.


Mild mitochondrial uncoupling as a therapeutic strategy.
Mild mitochondrial uncoupling, or the reduction of the efficiency of energy conversion without compromising intracellular high energy phosphate levels, is a protective therapeutic strategy under many laboratory conditions. Here we discuss these conditions, which include both cell and animal models of ischemia reperfusion and complications associated with the metabolic syndrome. We also discuss drugs that promote mild mitochondrial uncoupling and naturally occurring mild mitochondrial uncoupling pathways involving free fatty acid cycling and K(+) transport.


Chiral diamines 4: a computational study of the enantioselective deprotonation of Boc-pyrrolidine with an alkyllithium in the presence of a chiral diamine.
The enantioselective deprotonation of N-Boc-pyrrolidine (1) with i-PrLi-(-)-sparteine has been studied at theoretical levels up through B3P86/6-31G. Four low-energy intermediate complexes involving i-PrLi-(-)-sparteine and 1 were located via geometry optimizations; two of these complexes would lead to abstraction of the pro-S hydrogen from 1, and the other two complexes would lead to loss of the pro-R hydrogen. The lowest-energy intermediate complex was found to lead to loss of the pro-S hydrogen as observed experimentally. Transition states for the deprotonations were located using the synchronous transit-guided quasi-Newton method. The calculated activation enthalpy for transfer of the pro-S hydrogen within the lowest-energy intermediate complex, 10.8 kcal/mol, is reasonable for a reaction that occurs at a relatively low temperature, and the calculated kinetic hydrogen isotope effect is in agreement with experimental data. The lower enantioselectivity observed experimentally for deprotonation of 1 using t-BuLi-(-)-sparteine is attributed to a transition-state effect due to increased steric interaction engendered by the bulky t-BuLi. Replacement of the tert-butoxycarbonyl group in 1 by a methoxycarbonyl is predicted to result in a slower deprotonation with somewhat decreased enantioselectivity. Asymmetric deprotonation of 1 using i-PrLi in combination with the C(2)-symmetric diamine, (S,S)-1,2-bis(N,N-dimethylamino)cyclohexane, was calculated to be much less selective than is the deprotonation mediated by (-)-sparteine as observed experimentally. The relative energies of the intermediate complexes were fairly well-reproduced by ONIUM calculations in which the sparteine ligand less its nitrogen atoms was treated by molecular mechanics and the remainder of the complex was treated by quantum mechanics.


Susceptibility of normal human lymphocytes to infection with HTLV-III/LAV.
Normal human peripheral blood lymphocytes were tested for their susceptibility to infection with retroviruses isolated from patients with the acquired immunodeficiency syndrome (AIDS) or AIDS-related complex. Of 10 normal individuals tested, lymphocytes from all subjects became infected and produced virus as detected by assay for Mg+2-dependent reverse transcriptase. Lymphocytes from different individuals were demonstrated to be either high or low producers of reverse transcriptase after infection. The kinetics of virus production were similar in cells from both high- and low-producing individuals. A significant correlation was observed between high and low viral-producing lymphocytes and expression of the Leu-3/T4 (CD4) surface molecule. Mitogen-stimulated peripheral blood lymphocytes exposed to HTLV-III/LAV manifested productive viral infection, as reflected by the appearance of early syncytia, followed by reverse transcriptase. Unstimulated peripheral blood lymphocyte cultures displayed late syncytia but no detectable reverse transcriptase upon exposure to virus. The addition of anti-human interferon-alpha did not appear to have an appreciable effect on viral production in normal peripheral blood lymphocytes exposed to the virus.


Ginsenoside Rg1 promotes peripheral nerve regeneration in rat model of nerve crush injury.
Searching for effective drugs which are capable of promoting nerve regeneration after nerve injuries has gained extensive attention. Ginsenoside Rg1 (GRg1) is one of the bioactive compounds extracted from ginseng. GRg1 has been shown to be neuroprotective in many in vitro studies, which raises the possibility of using GRg1 as a neuroprotective agent after nerve injuries. However, such a possibility has never been tested in in vivo studies. The present study was designed to investigate the efficacy of GRg1 in promoting nerve regeneration after nerve crush injury in rats. All rats were randomly divided into four groups (n=8 in each group) after crush injury and were intraperitoneally administrated daily for 4 weeks with 1mg/kg, or 5mg/kg GRg1 (low or high dose GRg1 groups), or 100mug/kg mecobalamin or normal saline, respectively. The axonal regeneration was investigated by retrograde labeling and morphometric analysis. The motor functional recovery was evaluated by electrophysiological studies, behavioral tests and histological appearance of the target muscles. Our data showed that high dose GRg1 achieved better axonal regeneration and functional recovery than those achieved by low dose GRg1 and mecobalamin. The final outcome of low dose GRg1 and mecobalamin was similar in both morphological and functional items, which was significantly better than that in saline group. These findings show that GRg1 is capable of promoting nerve regeneration after nerve injuries, suggesting the possibility of developing GRg1 a neuroprotective drug for peripheral nerve repair applications.


Consumption of red-hot chili pepper increases symptoms in patients with acute anal fissures.
Red-hot chili pepper and other spices have been blamed for causing or exacerbating symptoms of anal pathologies like anal fissure and hemorrhoids. This study was aimed to determine if consumption of chilies increases symptoms of acute anal fissures. Individual patients were randomized to receive capsules containing chili or placebo for one week in addition to analgesics and fiber supplement. Patients were asked to note score for symptoms like pain, anal burning, and pruritus during the study period. After one week, cross over treatment was administered to the same group of patients with the same methodology and results were noted at the end of two weeks. Fifty subjects were recruited for this study 43 of them completed the trial (22 in the chili group and 21 in the placebo group). The daily mean pain score was significantly lower in the placebo group in the study period. [Score 2.05 in chili group and 0.97 in placebo group, p<0.001]. There was a significant burning sensation experienced by the patients in the chili group (score 1.85 for the chili group vs. 0.71 for the placebo group, p<0.001). Patient's mean recorded improvement score was significantly higher after taking placebo. 81.3% patients preferred placebo while 13.9% preferred chilies. Two patients had no preference. Consumption of chili does increase the symptoms of acute anal fissure and reduces patient compliance.


A mutation in the latency-related gene of bovine herpesvirus 1 disrupts the latency reactivation cycle in calves.
Bovine herpesvirus 1 (BHV-1) is an important pathogen of cattle, and infection is usually initiated via the ocular or nasal cavity. Following acute infection, the primary site for BHV-1 latency is the sensory neuron. Reactivation from latency occurs sporadically, resulting in virus shedding and transmission to uninfected cattle. The only abundant viral transcript expressed during latency is the latency-related (LR) RNA, suggesting that it mediates some aspect of latency. An LR mutant was constructed by inserting three stop codons near the beginning of the LR-RNA, suggesting that expression of LR proteins would be altered. The LR mutant grew with wild-type (wt) efficiency in bovine kidney cells (MDBK). When calves were infected with the LR mutant, a dramatic decrease (3 to 4 logs) in ocular, but not nasal, viral shedding occurred during acute infection relative to the wt or the LR-rescued virus (M. Inman, L. Lovato, A. Doster, and C. Jones, J. Virol. 75:8507-8515, 2001). In this study, we examined the latency reactivation cycle in calves infected with the LR mutant and compared these results to those from calves infected with wt BHV-1 or the LR-rescued virus. During acute infection, lower levels of infectious virus were detected in trigeminal ganglion homogenates from calves infected with the LR mutant. As judged by in situ hybridization, BHV-1-positive neurons were detected in trigeminal ganglia of calves infected with the wt but not the LR mutant. Although LR-RNA was detected by reverse transcription-PCR in calves latently infected with the LR mutant, a semiquantitative PCR analysis revealed that lower levels of viral DNA were present in trigeminal ganglia of calves infected with the LR mutant. Dexamethasone treatment of calves latently infected with wt BHV-1 or the LR-rescued virus, but not the LR mutant, consistently induced reactivation from latency, as judged by shedding of infectious virus from the nose or eyes and increases in BHV-1-specific antibodies. In summary, this study demonstrates that wt expression of LR gene products plays an important role in the latency reactivation cycle of BHV-1 in cattle.


Auxin Response Factors promote organogenesis by chromatin-mediated repression of the pluripotency gene SHOOTMERISTEMLESS.
Specification of new organs from transit amplifying cells is critical for higher eukaryote development. In plants, a central stem cell pool maintained by the pluripotency factor SHOOTMERISTEMLESS (STM), is surrounded by transit amplifying cells competent to respond to auxin hormone maxima by giving rise to new organs. Auxin triggers flower initiation through Auxin Response Factor (ARF) MONOPTEROS (MP) and recruitment of chromatin remodelers to activate genes promoting floral fate. The contribution of gene repression to reproductive primordium initiation is poorly understood. Here we show that downregulation of the STM pluripotency gene promotes initiation of flowers and uncover the mechanism for STM silencing. The ARFs ETTIN (ETT) and ARF4 promote organogenesis at the reproductive shoot apex in parallel with MP via histone-deacetylation mediated transcriptional silencing of STM. ETT and ARF4 directly repress STM, while MP acts indirectly, through its target FILAMENTOUS FLOWER (FIL). Our data suggest that - as in animals- downregulation of the pluripotency program is important for organogenesis in plants.


Diversification of sterol methyltransferase enzymes in plants and a role for β-sitosterol in oriented cell plate formation and polarized growth.
Phytosterols are classified into C24-ethylsterols and C24-methylsterols according to the different C24-alkylation levels conferred by two types of sterol methyltransferases (SMTs). The first type of SMT (SMT1) is widely conserved, whereas the second type (SMT2) has diverged in charophytes and land plants. The Arabidopsis smt2 smt3 mutant is defective in the SMT2 step, leading to deficiency in C24-ethylsterols while the C24-methylsterol pathway is unchanged. smt2 smt3 plants exhibit severe dwarfism and abnormal development throughout their life cycle, with irregular cell division followed by collapsed cell files. Preprophase bands are occasionally formed in perpendicular directions in adjacent cells, and abnormal phragmoplasts with mislocalized KNOLLE syntaxin and tubulin are observed. Defects in auxin-dependent processes are exemplified by mislocalizations of the PIN2 auxin efflux carrier due to disrupted cell division and failure to distribute PIN2 asymmetrically after cytokinesis. Although endocytosis of PIN2-GFP from the plasma membrane (PM) is apparently unaffected in smt2 smt3, strong inhibition of the endocytic recycling is associated with a remarkable reduction in the level of PIN2-GFP on the PM. Aberrant localization of the cytoplasmic linker associated protein (CLASP) and microtubules is implicated in the disrupted endocytic recycling in smt2 smt3. Exogenous C24-ethylsterols partially recover lateral root development and auxin distribution in smt2 smt3 roots. These results indicate that C24-ethylsterols play a crucial role in division plane determination, directional auxin transport, and polar growth. It is proposed that the divergence of SMT2 genes together with the ability to produce C24-ethylsterols were critical events to achieve polarized growth in the plant lineage.


Effect of Cobalt Precursors on Cobalt-Hydroxyapatite Used in Bone Regeneration and MRI.
In clinical dentistry practice, supplemental bone surgery or jawbone defect after tooth extraction must be assisted by a bone-filling material. Cobalt-substituted hydroxyapatite (COHA) effectively promotes bone cell growth, reduces the inflammatory response, and is an antibacterial agent. COHA can therefore be used as an alveolar bone-filling material or guided bone regeneration membrane. Meanwhile, COHA can be used in magnetic resonance imaging (MRI) with negative contrast agents and targeting materials without causing metal interference with the image. Hence, COHA has received increasing amounts of attention in recent years. However, the influence of different cobalt precursors on the synthesized COHA is still unknown. Therefore, COHA synthesized from 3 cobalt precursors (cobalt chloride, cobalt nitrate, and cobalt sulfate) was compared in this study. The results show that COHA synthesized by the precursor with the smallest anion radius, cobalt chloride, has a larger particle size (239 nm) and a higher cobalt ion substitution rate (15.6%). When the cobalt ion substitution rate increases, the MRI has a stronger contrast. Bioactivity data indicate that COHAC is more susceptible to degradation and therefore releases more cobalt ions to contribute to the differentiation of bone cells. Based on these studies, COHAC prepared with the cobalt chloride precursor has a higher cobalt ion substitution rate, faster degradation rate, better image contrast, and better bioactivity. It is therefore the preferred choice of bone-filling material for alveolar bone regeneration.


The endoplasmic reticulum mitochondrial calcium cross talk is downregulated in malignant pleural mesothelioma cells and plays a critical role in apoptosis inhibition.
The failure of apoptosis may contribute to the formation of cancer and to its resistance to therapy. Malignant pleural mesothelioma (MPM) is an aggressive tumor that responds poorly to standard chemo- and radio-therapies. Several studies have demonstrated that a plethora of oncogenes and tumor suppressors contribute to MPM onset/progression. Importantly, most of these genes are involved in the regulation of calcium (Ca2+)-handling. Cellular Ca2+ signaling is an important regulator of many physiological processes, and it has been widely reported to participate in the regulation of apoptotic cell death in cancer cells and tissues. However, in MPM the role of cellular Ca2+ has been poorly investigated. Therefore, we examined whether Ca2+ is involved in MPM. We found that mesothelioma cell lines and short-term cultures obtained from MPM-affected patients exhibited a critical dysregulation in Ca2+ signaling. We determined that this characteristic was associated with resistance to apoptotic stimuli and that correction of intracellular Ca2+ signaling resulted in the rescue of efficient apoptotic responses. In addition, we discovered that mitochondrial Ca2+-uptake plays a pivotal role as an inducer of apoptosis in MPM. Altogether, these findings suggest the identification of new MPM markers, which in turn could be potential targets for new therapeutic approaches.


A longitudinal analysis of the epidemiology and economic impact of inpatient admissions for chronic pancreatitis in the United States.
Chronic pancreatitis (CP) is a chronic, debilitating disorder associated with multiple complications, frequently necessitating hospitalization. The aim of this study was to investigate the longitudinal trends for hospitalization, mean length of stay (LOS), and cost associated with inpatient admissions for CP across the United States. Using the Nationwide Inpatient Sample, all hospitalizations between 1997 and 2014 were analyzed. We examined annual data for rate of hospitalization, average LOS and cost for CP inpatient admissions. Trends were described over the surveillance period. Between 1997 and 2014, the number of hospitalizations for patients with a primary discharge diagnosis of CP decreased by 41.5% (P<0.001). While the average LOS decreased by 21.2% from 6.2 days in 1997 to 4.9 days in 2014 (P<0.001), the mean charges for CP-related hospital admissions increased by 308.5% from $12,725 in 1997 to $39,260 (adjusted for inflation) in 2014 (P<0.001). The risk of a discharge for CP significantly increased from 1997-2014 for the 1-17 year age group (relative risk 1.518, 95% confidence interval 1.516-1.520; P<0.0001), while it significantly decreased over time for all the other age groups. Although it is reassuring that the average LOS has reduced, the cost associated with these hospitalizations has almost tripled. We postulate that the increase in cost is likely attributable to a greater number of studies and/or interventions. In order to deliver more cost-conscious care, further investigation is required into the effect that these additional investigations and interventions have on specific endpoints, including disease-specific and all-cause morbidity and mortality.


A randomized controlled pilot study to evaluate a technology platform for the assisted living of people with dementia and their carers.
The use of telemedicine is becoming increasingly popular in assisting with the home management of People with Dementia (PwD) by offering services to the carers that may enhance their ability to care for their relative for longer. A computerized platform, ALADDIN, was evaluated in its usefulness to reduce carer burden and distress and to improve their quality of life, in an attempt to delay institutionalization of PwD. ALADDIN offers educational material about dementia to carers and provides the opportunity to contact other carers and clinicians. ALADDIN also facilitates remote monitoring of the PwD and their carers by the clinicians to enable speedy delivery of appropriate intervention. The ALADDIN platform was piloted at three European sites, and used by thirty carers of PwD living in the community (platform group). The platform group and a control group of thirty PwD and their carers were assessed at baseline, 3 months, and 6 months. The results showed a significant improvement in the quality of life of the carers in the platform group, with some reduction in carer burden and distress. The platform was useful in monitoring the patients and facilitating contact with other professionals. Access to and use of the ALADDIN platform was rated positively by carers and clinicians. The ALADDIN platform's usefulness and applicability for prolonging the home management of PwD are discussed.


Selected Harmful and Potentially Harmful Constituents Levels in Commercial e-Cigarettes.
A broad range of commercially available electronic cigarette (e-cigarette) systems were tested for levels of emissions of harmful and potentially harmful constituents (HPHC), with a particular focus on the carbonyls: acetaldehyde, acrolein, and formaldehyde. The tobacco-specific nitrosamines N'-nitrosonornicotine and 4-(methylnitrosamino)-1-(3-bipyridyl)-1-butanone; the elements arsenic, cadmium, chromium, lead, and nickel; benzene; 1,3-butadiene; and benzo(a)pyrene were also quantified. The results show that except for the levels of carbonyls, all types of e-cigarettes performed in a similar manner, and emission levels for HPHCs were generally not quantifiable. However, levels of carbonyls, especially formaldehyde, were highly variable. Overall, the lowest levels of formaldehyde were observed in cartridge systems, which generally achieved substantial reductions in yields in comparison with cigarette smoke. Formaldehyde levels in open tank systems were variable; however, the median formaldehyde levels across different brands were substantially lower than the formaldehyde levels in cigarette smoke. The results for variable-power devices operated at the highest voltage confirmed existing literature data regardless of orientation and differences in puffing regimes. Furthermore, our results show that many products deliver consistent HPHC yields over a broad range of testing conditions (with minimal variability from one device to another, under a range of puffing conditions). However, some products exhibit high variability in emissions of HPHCs. The use of air blanks is further highlighted to assess nonproduct-related contributions to HPHC levels to avoid misrepresentation of the data. Overall, our results highlight that some but not all electronic cigarettes deliver low levels of carbonyls consistently across the full e-liquid depletion cycle under different test conditions. The need for further research and standardization work on assessment of variable-voltage electronic cigarettes is emphasized.


On-chip modulation of evanescent illumination and live-cell imaging with polymer waveguides.
Imaging of live cells was carried out using evanescent-wave excitation on a polymer waveguide chip. Integrated waveguide-based interferometric light modulators were fabricated in order to demonstrate on-chip control of excitation light, e.g., for time-lapse fluorescence microscopy. When combined with a sensitive high-resolution imaging system, the integrated waveguide-excitation platform provides an ideal method of near-surface studies of live cells, where photobleaching and/or phototoxicity effects are of critical concern.


Immunomodulatory metabolites released by the frog-killing fungus Batrachochytrium dendrobatidis.
Batrachochytrium dendrobatidis is a fungal pathogen in the phylum Chytridiomycota that causes the skin disease chytridiomycosis. Chytridiomycosis is considered an emerging infectious disease linked to worldwide amphibian declines and extinctions. Although amphibians have well-developed immune defenses, clearance of this pathogen from the skin is often impaired. Previously, we showed that the adaptive immune system is involved in the control of the pathogen, but B. dendrobatidis releases factors that inhibit in vitro and in vivo lymphocyte responses and induce lymphocyte apoptosis. Little is known about the nature of the inhibitory factors released by this fungus. Here, we describe the isolation and characterization of three fungal metabolites produced by B. dendrobatidis but not by the closely related nonpathogenic chytrid Homolaphlyctis polyrhiza. These metabolites are methylthioadenosine (MTA), tryptophan, and an oxidized product of tryptophan, kynurenine (Kyn). Independently, both MTA and Kyn inhibit the survival and proliferation of amphibian lymphocytes and the Jurkat human T cell leukemia cell line. However, working together, they become effective at much lower concentrations. We hypothesize that B. dendrobatidis can adapt its metabolism to release products that alter the local environment in the skin to inhibit immunity and enhance the survival of the pathogen.


Differential expression of the VLA family of integrins along the crypt-villus axis in the human small intestine.
Regulation of epithelial cell proliferation, migration and differentiation under physiological conditions remains poorly understood. Interaction of the cells with their underlying basement membrane through integrins, a specific subset of cell surface binding proteins, is one potential mechanism. In the present work, I examined this hypothesis by investigating the distribution of a variety of epithelial basement membrane proteins and the expression of the members of the VLA family of integrins in the adult intestinal epithelium. Indeed, this rapidly renewing simple epithelium contains within its functional unit, the crypt-villus axis, essentially two distinct cell populations: the proliferative and undifferentiated crypt cells and the mature enterocytes on the villus. Although immunolocalization of basement membrane molecules revealed that laminin, type IV collagen and heparan sulfate proteoglycan are distributed homogeneously all along the crypt-villus axis, other non-exclusive basement membrane components were found differentially expressed. Tenascin was concentrated at the base of both villus and lower crypt cells while cellular fibronectin was mostly detected in association with the crypt cells. Moreover, VLA beta 1 as well as 5 of the 6 VLA alpha subunits tested were expressed by intestinal epithelial cells under specific patterns of staining. The beta 1 and alpha 6 subunits were strongly detected at the base of all enterocytes while alpha 5, also detected all along the crypt-villus axis, was weaker and consistently appeared with a punctated/interrupted pattern. On the other hand, the VLA alpha 1, alpha 2 and alpha 3 were expressed at the basolateral domains of enterocytes under distinctive crypt-villus gradients. The alpha 1 subunit was detected at the base of all epithelial cells but lateral staining was only observed in differentiating cells (middle and upper crypt). Finally, in most specimens, alpha 2 and alpha 3 displayed strictly complementary staining patterns for the lower crypt region (alpha 2+, alpha 3-) and the upper crypt-to-villus region (alpha 2-, alpha 3+). Taken together, these data emphasize that proliferation, migration and differentiation in the normal state are susceptible to various influences including compositional changes in the basement membrane and differential expression of receptors for these components.


How Valuable are National Parks? Evidence from a Proposed National Park Expansion in Alaska.
When balancing environmental preservation and economic development, it is critical to evaluate how taxpayers value national park land and for what they are valuing it. One key component of this evaluation is to calculate a "passive use value," or the willingness to pay (WTP), for protection of land that may never directly be used, and to determine what benefits of land protection motivate this passive use value. We estimated the WTP for a 5% expansion of Denali National Park in Alaska (an expansion of 325,340 acres) using a questionnaire and the contingent valuation method. The survey first educated respondents on Alaskan geography, the current status of protected land and wildlife in Alaska, and common arguments for and against National Park expansion. The survey then described a proposal to expand the Denali National Park and asked a series of questions designed to bound the respondents WTP. Finally, the survey asked respondents to answer questions about what motivated their support or disapproval for the program. Multiple methodologies were used to estimate the national average WTP, resulting in a WTP range of a single payment of $77 to $409. This is estimated to be a total WTP of $15.1 billion to $79.3 billion for all individuals aged 18 to 64 in the United States, or $363.1 million to $1.91 billion per year in 2017 dollars. Respondents to the survey indicated that their support is driven primarily by increased protection from oil spills (85.8% deemed this very important or important), by increasing and protecting Alaska's biodiversity (84.3%), and preserving Alaska's beauty (84.2%). These results can help inform recent, current, and future debates about land use. President Obama significantly expanded land protections, while President Trump is continuing to scale back land protections, including opening up untouched land in Alaska in the Arctic National Wildlife Refuge for oil drilling. These results suggest that there are significant passive use values that should be considered when deciding whether to expand or reduce protection of land. Given that passive use values are significant, this suggests that the economic benefits must be large to tip a cost-benefit analysis in favor of reduced land protections.


Long-stay patients with and without intellectual disability in forensic psychiatric settings: comparison of characteristics and needs.
In recent years, concerns have been raised that too many patients stay for too long in forensic psychiatric services and that this is a particular problem in those with an intellectual disability. To compare the characteristics, needs, and care pathways of long-stay patients with and without intellectual disability within forensic psychiatric hospital settings in England. File reviews and questionnaires were completed for all long-stay patients in high secure and a representative sample of those in medium secure settings in England. Between-group analyses comparing patients with and without intellectual disability are reported. Of the 401 long-stay patients, the intellectual disability and non-intellectual disability groups were strikingly similar on many sociodemographic, clinical and forensic variables. The intellectual disability group had significantly lower lengths of stay, fewer criminal sections, restriction orders and prison transfers, and higher levels of behavioural incidents and risk assessment scores. In spite of similar offence histories and higher risk levels, those with intellectual disability appear to be diverted away from the criminal justice system and have shorter lengths of stay. This has implications about the applicability of the Transforming Care programme to this group.


Anti-endotoxin properties of cationic host defence peptides and proteins.
The innate immune system of mammals contains a series of peptides with overall positive charge and an amphipathic structure which have a variety of important properties in host defences. Although these are often termed cationic antimicrobial peptides, they have numerous roles in innate defences in all complex species of life and thus we prefer to refer to them as host defence peptides. These roles include: (i) an ability to kill micro-organisms directly, ranging from bacteria to viruses, fungi, parasites and helminths; (ii) an adjuvant activity in the adaptive response; and (iii) a multiplicity of roles in modulating innate immunity, including an apparent ability to stimulate protective innate immunity while suppressing harmful inflammatory/septic responses. This latter property may be one of the more important activities of these peptides in vivo. Innate immunity is thought to be triggered by the interaction of conserved bacterial components with particular receptors including Toll-like receptors (TLRs) on host cells. However, the initiation of the innate immune response through this route may trigger a pro-inflammatory cascade that is the principle cause of harmful conditions such as sepsis. Since we are exposed to potentially dangerous pathogens on a daily basis, the host response must contain certain checks and balances. We propose that host defence peptides have a role in feed-back modulation of inflammation under normal (low-pathogen exposure) conditions. This review surveys the available information regarding the anti-endotoxic/anti-inflammatory properties of host defence peptides, and will address whether this potential might be exploited for therapeutic benefit in sepsis.


Antibiotic use in acute respiratory infections and the ways patients pressure physicians for a prescription.
We identified those aspects of physician-patient communication that influence physicians to prescribe antibiotics for respiratory infections. A multimethod comparative case study was performed including descriptive field notes of outpatient visits. We included patients (children and adults) and clinicians in 18 purposefully selected family practices in a midwestern state. A total of 298 outpatient visits for acute respiratory tract (ART) infections were selected for analysis from more than 1600 encounters observed. Unnecessary antibiotic use and patterns of physician-patient communication were measured. Antibiotics were prescribed in 68% of the ART infection visits, and of those, 79% were determined to be unnecessary according to Centers for Disease Control and Prevention guidelines. Patients were observed to pressure physicians for medication. The types of patterns identified were direct request, candidate diagnosis (a diagnosis suggested by the patient), implied candidate diagnosis (a set of symptoms specifically indexing a particular diagnosis), portraying severity of illness, appealing to life-world circumstances, and previous use of antibiotics. Also, clinicians were observed to rationalize their antibiotic prescriptions by reporting medically acceptable reasons and diagnoses to patients. Patients strongly influence the antibiotic prescribing of physicians by using a number of different behaviors. To decrease antibiotic use for ART infections, patients should be educated about the dangers and limited benefits of such use, and clinicians should consider appropriate responses to these different patient pressures to prescribe antibiotics.


Amygdala activation during unconscious visual processing of food.
Hedonic or emotional responses to food have important positive and negative effects on human life. Behavioral studies have shown that hedonic responses to food images are elicited rapidly, even in the absence of conscious awareness of food. Although a number of previous neuroimaging studies investigated neural activity during conscious processing of food images, the neural mechanisms underlying unconscious food processing remain unknown. To investigate this issue, we measured neural activity using functional magnetic resonance imaging while participants viewed food and mosaic images presented subliminally and supraliminally. Conjunction analyses revealed that the bilateral amygdala was more strongly activated in response to food images than to mosaic images under both subliminal and supraliminal conditions. Interaction analyses revealed that the broad bilateral posterior regions, peaking at the posterior fusiform gyrus, were particularly active when participants viewed food versus mosaic images under the supraliminal compared with the subliminal condition. Dynamic causal modeling analyses supported the model in which the subcortical visual pathway from the pulvinar to the amygdala was modulated by food under the subliminal condition; in contrast, the model in which both subcortical and cortical (connecting the primary visual cortex, fusiform gyrus, and the amygdala) visual pathways were modulated by food received the most support under the supraliminal condition. These results suggest the possibility that unconscious hedonic responses to food may exert an effect through amygdala activation via the subcortical visual pathway.


Cytochrome P4501A biomarker indication of oil exposure in harlequin ducks up to 20 years after the Exxon Valdez oil spill.
Hydrocarbon-inducible cytochrome P4501A (CYP1A) expression was measured, as ethoxyresorufin-O-deethylase (EROD) activity, in livers of wintering harlequin ducks (Histrionicus histrionicus) captured in areas of Prince William Sound, Alaska, USA, oiled by the 1989 Exxon Valdez spill and in birds from nearby unoiled areas, during 2005 to 2009 (up to 20 years following the spill). The present work repeated studies conducted in 1998 that demonstrated that in harlequin ducks using areas that received Exxon Valdez oil, EROD activity was elevated nearly a decade after the spill. The present findings strongly supported the conclusion that average levels of hepatic EROD activity were higher in ducks from oiled areas than those from unoiled areas during 2005 to 2009. This result was consistent across four sampling periods; furthermore, results generated from two independent laboratories using paired liver samples from one of the sampling periods were similar. The EROD activity did not vary in relation to age, sex, or body mass of individuals, nor did it vary strongly by season in birds collected early and late in the winter of 2006 to 2007, indicating that these factors did not confound inferences about observed differences between oiled and unoiled areas. We interpret these results to indicate that harlequin ducks continued to be exposed to residual Exxon Valdez oil up to 20 years after the original spill. This adds to a growing body of literature suggesting that oil spills have the potential to affect wildlife for much longer time frames than previously assumed.


Expression of beta-actin during progression of mouse skin tumors.
Recombinant DNA techniques have been employed to isolate sequences that are over-expressed during multi-stage carcinogenesis in the mouse skin. One of these six cDNA sequences, mal-4, detected 1.9-kb transcripts that were expressed at levels 10-fold higher in squamous cell carcinomas (SCCs) in comparison to normal epidermis. A full-length cDNA for mal-4 was obtained from a lambda gt10 cDNA library made from an SCC-producing cell line, PDVC57. DNA sequencing of 1300 out of 1900 bp of the mal-4 cDNA and searching of Genbank showed greater than 95% DNA sequence similarity to mouse beta-actin over the entire cDNA and greater than 98% similarity over the 3' untranslated region, which is unique to the various isoforms of actins. Southern analysis showed no amplification or rearrangement of the beta-actin gene had occurred during tumor progression. The RNA:RNA hybrid protection assay was utilized to screen for the expression of mutated beta-actin(s) in mouse skin tumors. No evidence for mutation was obtained in any of the tumors examined. Fluorescence microscopy of tumor sections stained with rhodamine-conjugated phalloidin showed a peripheral pattern of F-actin localization with no gross differences between papillomas and carcinomas. Approximately equal amounts of beta-actin were observed following two-dimensional gel electrophoresis of proteins extracted from normal epidermis, papillomas or SCCs, indicating that the over-expression of beta-actin RNA in SCCs did not result in an increased steady-state level of beta-actin protein. These results suggest that alterations in actin metabolism accompany tumor progression.


An assay for cancer stem cell-induced angiogenesis on chick chorioallantoic membrane.
Angiogenesis is generally involved in tumor growth and metastasis. Cancer stem cells (CSCs) are considered to facilitate the angiogenesis. Therefore, CSCs could be the effective targets to stop angiogenesis. Recently, our group successfully generated CSC models from induced pluripotent stem cells (iPSCs) in the presence of conditioned medium derived from cancer derived cells. These novel model CSCs has been characterized by highly tumorigenic, angiogenic and metastatic potentials in vivo. The angiogenic potential of CSCs has been explained by the expression of both angiogenic factors and their receptors implying the angiogenesis in autocrine manner. In this protocol we optimized the method to evaluate tumor angiogenesis with the CSC model, which was described effective to assess sorafenib as an antiangiogenic drug, on chick chorioallantoic membrane (CAM) assay. Our results demonstrate that CSCs developed from iPSCs and CAM assay are a robust and cost-effective tool to evaluate tumor angiogenesis with CSCs. Collectively, CSCs in CAM assay could serve as a very useful model for the screening of potential therapeutic agents targeting tumor angiogenesis.


Electric field and current transport mechanisms in Schottky CdTe X-ray detectors under perturbing optical radiation.
Schottky CdTe X-ray detectors exhibit excellent spectroscopic performance but suffer from instabilities. Hence it is of extreme relevance to investigate their electrical properties. A systematic study of the electric field distribution and the current flowing in such detectors under optical perturbations is presented here. The detector response is explored by varying experimental parameters, such as voltage, temperature, and radiation wavelength. The strongest perturbation is observed under 850 nm irradiation, bulk carrier recombination becoming effective there. Cathode and anode irradiations evidence the crucial role of the contacts, the cathode being Ohmic and the anode blocking. In particular, under irradiation of the cathode, charge injection occurs and peculiar kinks, typical of trap filling, are observed both in the current-voltage characteristic and during transients. The simultaneous access to the electric field and the current highlights the correlation between free and fixed charges, and unveils carrier transport/collection mechanisms otherwise hidden.


Validation of SNP allele frequencies determined by pooled next-generation sequencing in natural populations of a non-model plant species.
Sequencing of pooled samples (Pool-Seq) using next-generation sequencing technologies has become increasingly popular, because it represents a rapid and cost-effective method to determine allele frequencies for single nucleotide polymorphisms (SNPs) in population pools. Validation of allele frequencies determined by Pool-Seq has been attempted using an individual genotyping approach, but these studies tend to use samples from existing model organism databases or DNA stores, and do not validate a realistic setup for sampling natural populations. Here we used pyrosequencing to validate allele frequencies determined by Pool-Seq in three natural populations of Arabidopsis halleri (Brassicaceae). The allele frequency estimates of the pooled population samples (consisting of 20 individual plant DNA samples) were determined after mapping Illumina reads to (i) the publicly available, high-quality reference genome of a closely related species (Arabidopsis thaliana) and (ii) our own de novo draft genome assembly of A. halleri. We then pyrosequenced nine selected SNPs using the same individuals from each population, resulting in a total of 540 samples. Our results show a highly significant and accurate relationship between pooled and individually determined allele frequencies, irrespective of the reference genome used. Allele frequencies differed on average by less than 4%. There was no tendency that either the Pool-Seq or the individual-based approach resulted in higher or lower estimates of allele frequencies. Moreover, the rather high coverage in the mapping to the two reference genomes, ranging from 55 to 284x, had no significant effect on the accuracy of the Pool-Seq. A resampling analysis showed that only very low coverage values (below 10-20x) would substantially reduce the precision of the method. We therefore conclude that a pooled re-sequencing approach is well suited for analyses of genetic variation in natural populations.


Interrogating Intracellular Zinc Chemistry with a Long Stokes Shift Zinc Probe ZincBY-4.
Previous work has shown that fluctuations in zinc content and subcellular localization play key roles in regulating cell cycle progression; however, a deep mechanistic understanding requires the determination of when, where, and how labile zinc pools are concentrated into or released from stores. Labile zinc ions can be difficult to detect with probes that require hydrolysis of toxic protecting groups or application at high concentrations that negatively impact cell function. We previously reported a BODIPY-based zinc probe, ZincBY-1, that can be used at working concentrations that are 20-200-fold lower than concentrations employed with other probes. To better understand how zinc pools can be visualized at such low probe concentrations, we modulated the photophysical properties via changes at the 5-position of the BODIPY core. One of these, ZincBY-4, exhibits an order of magnitude higher affinity for zinc, an 8-fold increase in brightness in response to zinc, and a 100 nm Stokes shift within cells. The larger Stokes shift of ZincBY-4 presents a unique opportunity for simultaneous imaging with GFP or fluorescein sensors upon single excitation. Finally, by creating a proxy for the cellular environment in spectrometer experiments, we show that the ZincBY series are highly effective at 50 nM because they can pass membranes and accumulate in regions of high zinc concentration within a cell. These features of the ZincBY probe class have widespread applications in imaging and for understanding the regulatory roles of zinc fluxes in live cells.


Discriminative stimulus effects of l-alpha-acetylmethadol (LAAM), buprenorphine and methadone in morphine-treated rhesus monkeys.
The discriminative stimulus effects of l-alpha-acetylmethadol (LAAM), l-alpha-acetylnormethadol (nor-LAAM), l-alpha-acetyldinormethadol (dinor-LAAM), buprenorphine and methadone were investigated in morphine-treated (3.2 mg/kg/day) rhesus monkeys (n = 3-6) discriminating between saline and naltrexone (0.01 mg/kg) and responding under a fixed ratio (FR) schedule of stimulus-shock termination. Monkeys responded on the naltrexone lever after either the administration of 0.01 mg/kg of naltrexone or the substitution of saline for the daily dose of morphine (i.e., 27-hr morphine deprived). Morphine dose-dependently reversed naltrexone lever responding in morphine-deprived monkeys. Methadone, LAAM, nor-LAAM and dinor-LAAM had morphine-like discriminative stimulus effects in all monkeys, whereas, buprenorphine had naltrexone-like discriminative stimulus effects in three monkeys and morphine-like effects in two monkeys; 24 hr after administration, buprenorphine antagonized the effects of morphine in the former and antagonized the effects of naltrexone in the latter. The agonist and antagonist effects of buprenorphine persisted for more than 6 days. The relative duration of action was: buprenorphine > LAAM > nor-LAAM = methadone = dinor-LAAM = morphine. That buprenorphine had markedly different discriminative stimulus effects in monkeys treated identically with morphine is likely due to the low efficacy of buprenorphine and emphasizes the difficulty in predicting the behavioral effects of buprenorphine in opioid-dependent individuals. The considerably longer duration of LAAM, than either nor-LAAM or dinor-LAAM, indicates that the rate of metabolite formation is important for the long duration of LAAM and further suggests that variations in metabolic activity among individuals might result in differences in the behavioral effects of LAAM.


Academic, social, and behavioral outcomes at age 12 of infants born preterm.
The effects of gradient levels of perinatal morbidity on school outcomes have been investigated at age 12 in four preterm groups, classified as healthy (no medical or neurological illness), medical morbidity, neurological morbidity, and small-for-gestational-age (SGA), and a full-term comparison group. Teachers report on academic competence, social skills, and problem behaviors. Data on school type, classroom setting, and school service use are gathered from school records. Preterm groups are found to be equivalent to full-term peers in social skills and problem behavior. Preterm groups with neurological and SGA morbidity have the lowest academic competence scores. Unexpectedly, preterm infants with medical morbidity have higher academic competence scores compared with the other preterm groups. School service use increases with greater perinatal morbidity and is contingent on multiple rather than single indicators of perinatal morbidity. Continued monitoring of preterm infants through early adolescence will ensure that appropriate school services and resources are available to maximize their school success.


Total body fat content and fat topography are associated differently with in vivo glucose metabolism in nonobese and obese nondiabetic women.
In this study, total body fat content and fat topography were related to glucose metabolism in the basal and insulin-stimulated states in 18 nonobese and 18 obese premenopausal nondiabetic women. All subjects received a euglycemic insulin (20 mU.min-1.m2) clamp study in combination with [3-3H]-D-glucose infusion and indirect calorimetry to quantitate total body glucose uptake, glucose oxidation, and nonoxidative glucose disposal. Total body fat content was determined with tritiated water, whereas body fat distribution was estimated from the WHR, the STR, and the VSR (measured by magnetic resonance imaging). In the postabsorptive state, total body glucose utilization, glucose oxidation, and nonoxidative glucose disposal rates were similar in nonobese and obese women, whereas during the insulin clamp all three metabolic parameters were reduced significantly in the obese group. In nonobese women, total body fat content was related inversely to both total and nonoxidative glucose disposal during the insulin clamp, whereas no relationship was found between glucose metabolism (total, oxidative, and nonoxidative) and WHR, STR, or VSR. In contrast, in obese women, no relationship was observed between total body fat content and any measure of insulin-mediated glucose metabolism. However, both WHR and VSR were related inversely to total, oxidative, and nonoxidative glucose disposal rates during the insulin clamp. These results suggest that total body fat content and body fat topography are associated differently with insulin-mediated glucose metabolism in nonobese and obese women. In the nonobese women, total body fat mass appears to be a primary determinant of tissue sensitivity to insulin, whereas in obese women, body fat topography exerts a more dominant effect.


Autopsy rate in suicide is low among elderly in Denmark compared with Finland.
National differences in the legislation on cause and manner of death investigation are reflected in a high autopsy rate in suicides in Finland and a low corresponding rate in Denmark. The consequences for mortality statistics of these different investigation practices on deaths classified as suicides in Denmark and Finland, respectively, are not known in detail. The aim of this article was to analyse autopsy rates in deaths classified as suicides, and to identify any differences in investigation practices in deaths with a comparable cause of death, but classified as unnatural deaths other than suicide. Data from the mortality registries were summarised for the years 2000, 2005 and 2010. Autopsy rates (total, forensic and medical) were analysed with regard to deaths classified as suicide, and they were compared for three age groups (1-50 years, 51-70 years and ≥71 years) and for causes of death. Deaths classified as suicide were compared with other unnatural classifications, and comparable causes of death were coded into six subgroups: poisonings, suffocations/strangulations, firearm discharges, drowning/submersions, explosions/flames and other/unspecified causes. The total autopsy rate for suicides was 99.8% in Finland and 13.2% in Denmark. Almost all of these autopsies were conducted as forensic autopsies. In the age group ≥71 years, Danish suicides outnumbered Finnish suicides (410 versus 283). The total autopsy rate was lower in the more senior age group in Denmark (19.5%, 9.9%, 5.6%), whereas it was consistently high in Finland (99.8%, 99.9%, 99.6%). Among Danish deaths due to poisonings, the autopsy rate was 89.5% when these were classified as accidents, but only 20.7% for cases classified as suicides. The number of deaths in the two Danish subgroups was comparable (550 versus 553). In Denmark, the decision regarding the need, if any, for a forensic autopsy is made during the external forensic examination of the body. Our study showed that the limited use of forensic autopsy to confirm the cause of death in deaths classified as suicides raises doubts about the accuracy of the Danish suicide mortality statistics. Our finding is emphasised by those cases in which the cause of death was registered as intentional self-poisoning. The high number of suicides among the elderly in Denmark is striking and begs further investigation and research. Overall, our data from Finland and Denmark reveal striking differences between the two countries and warrant further comparative studies on the subject in other countries.


Opposing microtubule motors control motility, morphology and cargo segregation during ER-to-Golgi transport.
We recently demonstrated that dynein and kinesin motors drive multiple aspects of endosomal function in mammalian cells. These functions include driving motility, maintaining morphology (notably through providing longitudinal tension to support vesicle fission), and driving cargo sorting. Microtubule motors drive bidirectional motility during traffic between the endoplasmic reticulum (ER) and Golgi. Here, we have examined the role of microtubule motors in transport carrier motility, morphology, and domain organization during ER-to-Golgi transport. We show that, consistent with our findings for endosomal dynamics, microtubule motor function during ER-to-Golgi transport of secretory cargo is required for motility, morphology, and cargo sorting within vesicular tubular carriers en route to the Golgi. Our data are consistent with previous findings that defined roles for dynein-1, kinesin-1 (KIF5B) and kinesin-2 in this trafficking step. Our high resolution tracking data identify some intriguing aspects. Depletion of kinesin-1 reduces the number of motile structures seen, which is in line with other findings relating to the role of kinesin-1 in ER export. However, those transport carriers that were produced had a much greater run length suggesting that this motor can act as a brake on anterograde motility. Kinesin-2 depletion did not significantly reduce the number of motile transport carriers but did cause a similar increase in run length. These data suggest that kinesins act as negative regulators of ER-to-Golgi transport. Depletion of dynein not only reduced the number of motile carriers formed but also caused tubulation of carriers similar to that seen for sorting nexin-coated early endosomes. Our data indicated that the previously observed anterograde-retrograde polarity of transport carriers in transit to the Golgi from the ER is maintained by microtubule motor function.


MeCP2 behaves as an elongated monomer that does not stably associate with the Sin3a chromatin remodeling complex.
MeCP2 is a transcription factor that recognizes and binds symmetrically methylated CpG dinucleotides to repress transcription. MeCP2 can associate with the Sin3a/histone deacetylase corepressor complex and mediate repression in a histone deacetylase-dependent manner. In extracts from rodent tissues, cultured cells, and Xenopus laevis oocytes, we find that only a small amount of mammalian MeCP2 interacts with Sin3a and that this interaction is not stable. Purification of rat brain MeCP2 (53 kDa) indicates no associated proteins despite an apparent molecular mass by size exclusion chromatography of 400-500 kDa. Biophysical analysis demonstrated that the large apparent size was not because of homo-multimerization, as MeCP2 consistently behaves as a monomeric protein that has an elongated shape. Our findings indicate the MeCP2 is not an obligate component of the Sin3a corepressor complex and may therefore engage a more diverse range of cofactors for repressive function.


Monoclonal antibody to human platelet glycoprotein I. II. Effects on human platelet function.
The effect on platelet function of a monoclonal platelet antibody to platelet membrane glycoprotein I was tested. This antibody, AN51, inhibited ristocetin or bovine factor VIII-induced aggregation but did not modify ADP, collagen type I or type III, thrombin or arachidonic acid induced aggregations. Furthermore, the adhesion-aggregation of platelets induced by microfibrils was also inhibited by the antibody. Platelet adhesion to rabbit aorta subendothelium was impaired by the antibody. The persistent adhesion of platelets to collagenase-treated subendothelium was also inhibited. These findings strongly suggested that platelet membrane glycoprotein I could interact with a non-collagenic microfibrillar component of subendothelium. The binding of factor VIII/von Willebrand factor to platelet membrane in the presence of ristocetin was decreased in the binding site for factor VIII/von Willebrand factor to allow platelet adhesion to subendothelium.


High-Risk Types of Human Papilloma Virus DNA Testing in Women with False Negative Cytology.
To determine whether high-risk types of human papilloma virus (hrHPV) DNA testing is reliable for selection patients in need of further investigation with colposcopy in women with increased risk of high-grade cervical lesions as a result of false negative cytology. The secondary objective was to compare the sensitivity of hrHPV testing on self-collected versus physician-collected samples for the detection of histological high-grade cervical intraepithelial neoplasia (CIN2+). Sixty-three patients identified with a missed abnormality following the re-evaluation of benign cervical cytology were included. A patient-collected and a physician-collected sample for HPV, colposcopy and cervical specimen collection for histology and cytology were performed. The sensitivity of hrHPV testing of physician-collected samples for CIN2+ was 100% (95% CI 82.7-100), and the negative predictive value (NPV) was 100% (95% CI 93.3-100). The sensitivity of the self-sampling device to identify CIN2+ was 84.6% (95% CI 59.1-96.7), and the NPV was 94.4% (95% CI 83.4-98.8). The differences in the sensitivity and NPV between the 2 methods were non-significant. The agreement between the 2 methods regarding the HPV results was good, with a kappa value of 0.74 (95% CI 0.57-0.91). The current findings indicate that physician-collected samples for hrHPV DNA testing may be used as triage for the colposcopy of women with false negative cytology.


Coupling Genome-wide Transcriptomics and Developmental Toxicity Profiles in Zebrafish to Characterize Polycyclic Aromatic Hydrocarbon (PAH) Hazard.
Polycyclic Aromatic Hydrocarbons (PAHs) are diverse environmental pollutants associated with adverse human health effects. Many studies focus on the carcinogenic effects of a limited number of PAHs and there is an increasing need to understand mechanisms of developmental toxicity of more varied yet environmentally relevant PAHs. A previous study characterized the developmental toxicity of 123 PAHs in zebrafish. Based on phenotypic responses ranging from complete inactivity to acute mortality, we classified these PAHs into eight bins, selected 16 representative PAHs, and exposed developing zebrafish to the concentration of each PAH that induced 80% phenotypic effect. We conducted RNA sequencing at 48 h post fertilization to identify gene expression changes as a result of PAH exposure. Using the Context Likelihood of Relatedness algorithm, we inferred a network that links the PAHs based on coordinated gene responses to PAH exposure. The 16 PAHs formed two broad clusters: Cluster A was transcriptionally more similar to the controls, while Cluster B consisted of PAHs that were generally more developmentally toxic, significantly elevated cyp1a transcript levels, and induced Ahr2-dependent Cyp1a protein expression in the skin confirmed by gene-silencing studies. We found that cyp1a transcript levels were associated with transcriptomic response, but not with PAH developmental toxicity. While all cluster B PAHs predominantly activated Ahr2, they also each enriched unique pathways like ion transport signaling, which likely points to differing molecular events between the PAHs downstream of Ahr2. Thus, using a systems biology approach, we have begun to evaluate, classify, and define mechanisms of PAH toxicity.


Nonrandom turnover of actin and tubulin in cultured rabbit cardiac fibroblasts.
Total protein fractional rates of growth, synthesis, and degradation were assessed in primary cultures of rabbit cardiac fibroblasts. Differences in fractional growth rates were produced by subculturing cells at low density and growing them to confluence. Total protein fractional degradative rates were then derived by subtracting fractional growth rates from measured fractional synthetic rates (obtained in [3H]leucine pulse-labeling experiments). Actin and tubulin degradation were studied in similar rapidly and slowly growing cultures. [35S]methionine pulse-chase experiments, followed by dodecyl sulfate-polyacrylamide gel electrophoresis, fluorography, and densitometry were used to determine the amount of labeled actin and tubulin remaining in cultures at various times during the chase (0-96 h). The indirect study showed a substantially lower total protein fractional degradative rate during rapid vs. slow growth (0.04 +/- 0.13 vs. 0.42 +/- 0.01 d-1 at 2 and 15 days after subculture, respectively; P less than 0.01). At both growth rates, the disappearance of labeled actin and tubulin was delayed, suggesting a more complex model for their degradation than random decay. Serum deprivation of slowly growing fibroblasts increased the rate of disappearance of both proteins by eliminating the delay in their breakdown. Thus the suppression of protein degradation during rapid growth appears to result from the presence of relatively greater amounts of "new" actin and tubulin (and possible other long-lived proteins) that are kinetically distinct from the total intracellular pools of these proteins with respect to their susceptibility to proteolysis.


Enhanced age-dependent cerebrovascular dysfunction is mediated by adaptor protein p66Shc.
Aging is an independent risk factor for cardiovascular and cerebrovascular disease. To date, little is known about the mechanisms of aging of cerebral arteries and whether the aging gene p66(Shc) is implicated in it. The present study was designed to assess age-induced vascular dysfunction in cerebral and systemic arteries of wild type (wt) and p66(Shc-/-) mice. Basilar arteries and size matched second order femoral arteries of 3-month (3M), 6-month (6M) and 2-year old (2Y) mice were studied in wt and p66(Shc-/-) mice. To assess vascular function, arterial rings mounted in a myograph for isometric tension recordings were exposed to increasing concentrations of acetylcholine and sodium nitroprusside. Reactive oxygen species (ROS) generation was assessed in femoral and basilar arteries using the spin trap 1-hydroxy-3-methoxycarbonyl-2,2,5,5-tetramethyl-pyrrolidine. In wt mice, endothelial function of the femoral artery was not affected by age unlike in the basilar artery where an age-dependent dysfunction was observed. In p66(Shc-/-) a similar response was observed in the femoral artery; however, age-dependent endothelial dysfunction of the basilar artery was blunted as compared to wt. Levels of ROS were comparable in the femoral arteries of 3M and 2Y of wt and p66(Shc-/-) mice. Differently, ROS levels in the basilar artery of wt mice were strongly increased by age unlike in p66(Shc-/-) mice where they remained comparable irrespective of age. Endothelial function in cerebral arteries, but not in size-matched systemic ones, is heavily impaired by aging. This process is paralleled by an increased ROS production and is mediated by the p66(Shc) gene.


Crystal structure of defensin HNP-3, an amphiphilic dimer: mechanisms of membrane permeabilization.
Defensins (molecular weight 3500 to 4000) act in the mammalian immune response by permeabilizing the plasma membranes of a broad spectrum of target organisms, including bacteria, fungi, and enveloped viruses. The high-resolution crystal structure of defensin HNP-3 (1.9 angstrom resolution, R factor 0.19) reveals a dimeric beta sheet that has an architecture very different from other lytic peptides. The dimeric assembly suggests mechanisms by which defensins might bind to and permeabilize the lipid bilayer.


The difference in patterns of motor and cognitive function in chronic fatigue syndrome and severe depressive illness.
Chronic fatigue syndrome (CFS) and major depressive disorder (MDD) share many symptoms and aetiological factors but may have different neurobiological underpinnings. We wished to determine the profile of the biological variables disturbed in CFS and MDD, and identify any critical factors that differentiate the disorders. Thirty patients with CFS, 20 with MDD and 15 healthy controls matched group-wise for age and sex were recruited. Subjects were given a detailed battery of motor and cognitive tests, including measures of psychomotor speed, memory and maximal voluntary muscle contraction in both the morning and evening that were balanced to avoid order effects. CFS patients generally performed worse on cognitive tests than healthy controls, but better than patients with MDD. Both patient groups had markedly impaired motor function compared with healthy controls. MDD subjects showed a significantly greater diurnal improvement in maximal voluntary contraction than healthy controls. Patients with CFS and MDD show similarly substantial motor impairment, but cognitive deficits are generally more marked in MDD. Diurnal changes in some functions in MDD may differentiate the disorder from CFS.


Eosinophil-mediated signalling attenuates inflammatory responses in experimental colitis.
Eosinophils reside in the colonic mucosa and increase significantly during disease. Although a number of studies have suggested that eosinophils contribute to the pathogenesis of GI inflammation, the expanding scope of eosinophil-mediated activities indicate that they also regulate local immune responses and modulate tissue inflammation. We sought to define the impact of eosinophils that respond to acute phases of colitis in mice. Acute colitis was induced in mice by administration of dextran sulfate sodium, 2,4,6-trinitrobenzenesulfonic acid or oxazolone to C57BL/6J (control) or eosinophil deficient (PHIL) mice. Eosinophils were also depleted from mice using antibodies against interleukin (IL)-5 or by grafting bone marrow from PHIL mice into control mice. Colon tissues were collected and analysed by immunohistochemistry, flow cytometry and reverse transcription PCR; lipids were analysed by mass spectroscopy. Eosinophil-deficient mice developed significantly more severe colitis, and their colon tissues contained a greater number of neutrophils, than controls. This compensatory increase in neutrophils was accompanied by increased levels of the chemokines CXCL1 and CXCL2, which attract neutrophils. Lipidomic analyses of colonic tissue from eosinophil-deficient mice identified a deficiency in the docosahexaenoic acid-derived anti-inflammatory mediator 10, 17- dihydroxydocosahexaenoic acid (diHDoHE), namely protectin D1 (PD1). Administration of an exogenous PD1-isomer (10S, 17S-DiHDoHE) reduced the severity of colitis in eosinophil-deficient mice. The PD1-isomer also attenuated neutrophil infiltration and reduced levels of tumour necrosis factor-α, IL-1β, IL-6 and inducible NO-synthase in colons of mice. Finally, in vitro assays identified a direct inhibitory effect of PD1-isomer on neutrophil transepithelial migration. Eosinophils exert a protective effect in acute mouse colitis, via production of anti-inflammatory lipid mediators.


Transcranial direct current stimulation lessens dual task cost in people with Parkinson's disease.
Parkinson's disease (PD) progressively impairs motor and cognitive function. Gait dysfunction in PD is exacerbated during dual task gait. Transcranial direct current stimulation (tDCS) may therapeutically benefit motor and cognitive deficits. We examined the effect of a bilateral tDCS protocol on dual task gait in people with PD. Participants with PD between 50 and 80 years received two sessions of tDCS protocol (1 active, 1 sham) separated by 7days. tDCS protocols were randomized and blinded to participants. After each tDCS protocol, participants performed single and dual task gait. Single 20-min session of bilateral tDCS (dorsolateral prefrontal cortex; left=anode, right=cathode) at 2mA and one sham session. Each participant was assessed at baseline for disease severity [Unified Parkinson's Disease Rating Scale (UPDRS)] and executive function [Repeatable Battery for the Assessment of Neuropsychological Status (RBANS)]. Following each tDCS condition (active and sham), participants performed Timed Up and Go (TUG) single and dual task conditions (TUGalone, TUGmotor, TUGcognitive) and PDQ-39. Ten participants average age of 68.7 years (±10.2) and average PD duration average 7.9 years (±7.1) were included. The UPDRS (M=37) and RBANS (M=13%ile) were administered prior to testing. No differences were observed on dependent t-test for TUG conditions or PDQ-39. Dual task cost TUGmotor was -20.95% (tDCSactive) versus -22.58% (tDCSsham) and TUGcognitive was -25.24% (tDCSactive) versus -41.85% (tDCSsham). Our bilateral tDCS protocol in people with PD did not significantly improve dual task gait. However, dual task cost following tDCS was lessened, most dramatically in the presence of a cognitive distractor. A larger sample size is warranted to draw further conclusions about our bilateral tDCS approach.


Transmembrane protein rotaxanes reveal kinetic traps in the refolding of translocated substrates.
Understanding protein folding under conditions similar to those found in vivo remains challenging. Folding occurs mainly vectorially as a polypeptide emerges from the ribosome or from a membrane translocon. Protein folding during membrane translocation is particularly difficult to study. Here, we describe a single-molecule method to characterize the folded state of individual proteins after membrane translocation, by monitoring the ionic current passing through the pore. We tag both N and C termini of a model protein, thioredoxin, with biotinylated oligonucleotides. Under an electric potential, one of the oligonucleotides is pulled through a α-hemolysin nanopore driving the unfolding and translocation of the protein. We trap the protein in the nanopore as a rotaxane-like complex using streptavidin stoppers. The protein is subjected to cycles of unfolding-translocation-refolding switching the voltage polarity. We find that the refolding pathway after translocation is slower than in bulk solution due to the existence of kinetic traps.


Molecular cloning, genomic organization, promoter activity, and tissue-specific expression of the mouse ryudocan gene.
Ryudocan, a ubiquitous heparan sulfate proteoglycan, is a member of the syndecan family of cell surface proteoglycans. The full-length cDNA encoding the murine ryudocan core protein has now been cloned and sequenced. The deduced primary structure of mouse ryudocan, including the three glycosaminoglycan attachment sites in the extracellular domain as well as the transmembrane and cytoplasmic regions, is highly similar to those of the rat, human, and chicken proteins. Northern analysis detected a 2.7-kb transcript in all mouse tissues examined, with the highest concentrations apparent in liver, kidney, and lung. The mouse ryudocan gene was shown to span approximately 19.7 kb of genomic DNA and to contain five exons, with an intron-exon organization identical to that of the human gene. The promoter region of the mouse gene contains various cis-acting elements, including a TATA-like box and a GC box as well as potential binding sites for the transcription factors NF-IL6, MyoD, GATA, C/EBP, AP-2, NF-kappaB, AP-1, and Sp1. Transient transfection experiments with a construct containing the 690 bp upstream of the transcription start site fused to a luciferase reporter gene showed functional promoter activity. Deletion analysis suggested that the proximal promoter region including the TATA-like box, the GC box, and other Sp1 binding sites was required for full transcriptional activity. These findings will be useful for the study of ryudocan gene regulation and the generation of mice with targeted disruption of the gene.


Automated suppression of sample-related artifacts in Fluorescence Correlation Spectroscopy.
Fluorescence Correlation Spectroscopy (FCS) in cells often suffers from artifacts caused by bright aggregates or vesicles, depletion of fluorophores or bleaching of a fluorescent background. The common practice of manually discarding distorted curves is time consuming and subjective. Here we demonstrate the feasibility of automated FCS data analysis with efficient rejection of corrupted parts of the signal. As test systems we use a solution of fluorescent molecules, contaminated with bright fluorescent beads, as well as cells expressing a fluorescent protein (ICA512-EGFP), which partitions into bright secretory granules. This approach improves the accuracy of FCS measurements in biological samples, extends its applicability to especially challenging systems and greatly simplifies and accelerates the data analysis.


Cutting edge: tumor-targeting antibodies enhance NKG2D-mediated NK cell cytotoxicity by stabilizing NK cell-tumor cell interactions.
Monoclonal antibodies represent a promising approach to fight a variety of tumors, but their mode of action remains to be fully understood. NK cells can recognize Ab-coated targets, as well as stress ligands, on tumor cells. In this study, we investigated how NK cells integrate both kinds of activating signals. NK cell-mediated killing was maximal with the combined recognition of NKG2D ligands and Ab; surprisingly, only NKG2D engagement substantially enhanced degranulation. Conversely, Ab recognition by NK cells uniquely increased contact stability with tumor cells. Furthermore, using intravital imaging of solid tumors, we showed that Ab recognition favored prolonged interactions between NK cells and targets. Altogether, our results demonstrate that NK cell-mediated killing can be differentially regulated at the level of degranulation and contact stability by distinct activating receptors. Thus, complementary signals mediated by recognition of stress ligands and tumor-specific Abs may contribute to the efficacy of NK cells during mAb therapy.


Racial/ethnic differences in responses to the everyday discrimination scale: a differential item functioning analysis.
The authors examined the impact of race/ethnicity on responses to the Everyday Discrimination Scale, one of the most widely used discrimination scales in epidemiologic and public health research. Participants were 3,295 middle-aged US women (African-American, Caucasian, Chinese, Hispanic, and Japanese) from the Study of Women's Health Across the Nation (SWAN) baseline examination (1996-1997). Multiple-indicator, multiple-cause models were used to examine differential item functioning (DIF) on the Everyday Discrimination Scale by race/ethnicity. After adjustment for age, education, and language of interview, meaningful DIF was observed for 3 (out of 10) items: "receiving poorer service in restaurants or stores," "being treated as if you are dishonest," and "being treated with less courtesy than other people" (all P's < 0.001). Consequently, the "profile" of everyday discrimination differed slightly for women of different racial/ethnic groups, with certain "public" experiences appearing to have more salience for African-American and Chinese women and "dishonesty" having more salience for racial/ethnic minority women overall. "Courtesy" appeared to have more salience for Hispanic women only in comparison with African-American women. Findings suggest that the Everyday Discrimination Scale could potentially be used across racial/ethnic groups as originally intended. However, researchers should use caution with items that demonstrated DIF.


Covariation in life-history traits: differential effects of diet on condition, hormones, behavior, and reproduction in genetic finch morphs.
The relative contribution of genetic and environmental factors in determining variation in life-history traits is of central interest to evolutionary biologists, but the physiological mechanisms underlying these traits are still poorly understood. Here we experimentally demonstrate opposing effects of nutritional stress on immune function, endocrine physiology, parental care, and reproduction between red and black head-color morphs of the Gouldian finch (Erythrura gouldiae). Although the body condition of black morphs was largely unaffected by diet manipulation, red birds were highly sensitive to dietary changes, exhibiting considerable within-individual changes in condition and immune function. Consequently, nutritionally stressed red birds delayed breeding, produced smaller broods, and reared fewer and lower-quality foster offspring than black morphs. Differences in offspring quality were largely due to morph-specific differences in parental effort: red morphs reduced parental provisioning, whereas black morphs adaptively elevated their provisioning effort to meet the increased nutritional demands of their foster brood. Nutritionally stressed genetic morphs also exhibited divergent glucocorticoid responses. Black morphs showed reduced corticosterone-binding globulin (CBG) concentrations and increased levels of free corticosterone, whereas red morphs exhibited reduced free corticosterone levels and elevated CBG concentrations. These opposing glucocorticoid responses highlight intrinsic differences in endocrine sensitivities and plasticity between genetic morphs, which may underlie the morph-specific differences in condition, behavior, and reproduction and thus ultimately contribute to the evolution and maintenance of color polymorphism.


GeneORator: An Effective Strategy for Navigating Protein Sequence Space More Efficiently through Boolean OR-Type DNA Libraries.
Directed evolution requires the creation of genetic diversity and subsequent screening or selection for improved variants. For DNA mutagenesis, conventional site-directed methods implicitly utilize the Boolean AND operator (creating all mutations simultaneously), producing a combinatorial explosion in the number of genetic variants as the number of mutations increases. We introduce GeneORator, a novel strategy for creating DNA libraries based on the Boolean logical OR operator. Here, a single library is divided into many subsets, each containing different combinations of the desired mutations. Consequently, the effect of adding more mutations on the number of genetic combinations is additive (Boolean OR logic) and not exponential (AND logic). We demonstrate this strategy with large-scale mutagenesis studies, using monoamine oxidase-N ( Aspergillus niger) as the exemplar target. First, we mutated every residue in the secondary structure-containing regions (276 out of a total 495 amino acids) to screen for improvements in kcat. Second, combinatorial OR-type libraries permitted screening of diverse mutation combinations in the enzyme active site to detect activity toward novel substrates. In both examples, OR-type libraries effectively reduced the number of variants searched up to 1010-fold, dramatically reducing the screening effort required to discover variants with improved and/or novel activity. Importantly, this approach enables the screening of a greater diversity of mutation combinations, accessing a larger area of a protein's sequence space. OR-type libraries can be applied to any biological engineering objective requiring DNA mutagenesis, and the approach has wide ranging applications in, for example, enzyme engineering, antibody engineering, and synthetic biology.


Systematic review of AAC intervention research for adolescents and adults with autism spectrum disorder.
Much of augmentative and alternative communication (AAC) research for individuals with autism spectrum disorder has focused on young children. Given that the lives, communication, strengths, and needs of adolescents and adults with autism spectrum disorder are quite different from those of young children, the purpose of the current study was to consolidate current AAC intervention research findings specific to these individuals. A systematic review was conducted to identify and evaluate relevant research. Results indicate that AAC intervention benefits adolescents and adults with autism spectrum disorder. However, more research is urgently needed. Future research focused on supporting communicative functions other than requesting (e.g., social closeness, information transfer) while participating in contexts important to the lives of adolescents and adults may be particularly valuable.


The psychological and legal aftermath of false arrest and imprisonment.
False arrest and imprisonment can be an extraordinarily stressful psychological trauma. This is clearly demonstrated in the evaluation of forensic cases alleging false arrest and imprisonment, a review of the recent forensic psychiatric literature and reported legal cases. A clinical vignette is presented that illustrates the psychological trauma and sequelae associated with false arrest and imprisonment. Psychiatric treatment of these individuals is discussed. A number of these cases are litigated.


Distinct associations of insula and cingulate volume with the cognitive and affective dimensions of alexithymia.
Alexithymia ("no words for feelings") is a major risk factor for psychosomatic and psychiatric conditions characterized by affect dysregulation. The alexithymia personality construct comprises an affective dimension, the level of subjective emotional experience (emotionalizing and fantasizing), and a cognitive dimension, referring to the cognitive control of emotions (identifying, analyzing, and verbalizing feelings). These two dimensions may differentially put individuals at risk for psychopathology, but their specific neural bases have rarely been investigated. Therefore, the aim of the present study was to find out whether the two alexithymia dimensions are associated with discriminable neural correlates. By means of voxel-based morphometry (VBM), differences in gray matter volumes were compared between 20 (10 male) high-scorers and 20 (9 male) low-scorers on the Toronto Alexithymia Scale (TAS-20), reflecting the cognitive alexithymia dimension. In a subset of 32 subjects, the impact of the affective alexithymia dimension was tested in addition, as assessed with the affective subscale of the Bermond-Vorst Alexithymia Questionnaire (BVAQ). Analysis 1 (cognitive alexithymia dimension) revealed significantly larger gray matter volumes in the right posterior insula in high-scorers compared to low-scorers on the TAS-20. Analysis 2 (affective alexithymia dimension) revealed that the affective alexithymia dimension, specifically the emotionalizing factor indicative of low emotional reactivity, was associated with larger gray matter volumes of the right cingulate cortex. These results suggest that the two alexithymia dimensions are associated with distinct structural correlates.


Microbiota-Driven Tonic Interferon Signals in Lung Stromal Cells Protect from Influenza Virus Infection.
Type I interferon (IFNα/β) pathways are fine-tuned to elicit antiviral protection while minimizing immunopathology; however, the initiating stimuli, target tissues, and underlying mechanisms are unclear. Using models of physiological and dysregulated IFNα/β receptor (IFNAR1) surface expression, we show here that IFNAR1-dependent signals set the steady-state IFN signature in both hematopoietic and stromal cells. Increased IFNAR1 levels promote a lung environment refractory to early influenza virus replication by elevating the baseline interferon signature. Commensal microbiota drive the IFN signature specifically in lung stroma, as shown by antibiotic treatment and fecal transplantation. Bone marrow chimera experiments identify lung stromal cells as crucially important for early antiviral immunity and stroma-immune cell interaction for late antiviral resistance. We propose that the microbiota-driven interferon signature in lung epithelia impedes early virus replication and that IFNAR1 surface levels fine-tune this signature. Our findings highlight the interplay between bacterial and viral exposure, with important implications for antibiotic use.


Corticotropin releasing hormone receptor alterations elicited by acute and chronic unpredictable stressor challenges in stressor-susceptible and resilient strains of mice.
Stressors increase corticotropin releasing hormone (CRH) functioning in hypothalamic and frontal cortical brain regions, and thus may contribute to the provocation of anxiety and depressive disorder. As the effects of stressors on these behavioral changes are more pronounced in some strains of mice (e.g., BALB/cByJ) than in others (e.g., C57BL/6ByJ), the present investigation assessed whether acute and chronic stressors would differentially influence CRH receptor immunoreactivity (ir-CRHr) and CRH receptor mRNA expression (CRH(1) and CRH(2)) in the orbital frontal cortex (OFC) of these strains. An acute noise stressor, and to a greater extent a chronic, variable stressor regimen reduced ir-CRHr in BALB/cByJ mice. In contrast, in the hardier C57BL/6ByJ mice the acute stressor increased ir-CRHr in portions of the OFC, whereas a chronic stressor tended to reduce ir-CRHr. However, whereas the acute stressor did not influence CRH(1) mRNA expression, the chronic stressor increased CRH(1) mRNA expression in both mouse strains. The CRH(2) expression appeared in low abundance in both strains and was unaltered by the stressor. In addition to the OFC variations, quantitative immunohistochemistry indicated that the chronic stressor treatment increased CRH immunoreactivity in the median eminence of C57BL/6ByJ mice, but co-expression of CRH and arginine vasopressin (AVP) immunoreactivity was not provoked by the stressors. The data support the view that stressors provoke marked variations of ir-CRHr in the OFC that might contribute to the differential anxiety/depression-like profiles ordinarily apparent in the stressor-vulnerable and -resilient mouse strains.


Contributions of the TEL-patch amino acid cluster on TPP1 to telomeric DNA synthesis by human telomerase.
Telomere maintenance is a highly coordinated process, and its misregulation is linked to cancer and telomere-shortening syndromes. Recent studies have shown that the TEL-patch--a cluster of amino acids on the surface of the shelterin component TPP1--is necessary for the recruitment of telomerase to the telomere in human cells. However, there has been only basic biochemical analysis of the role of TPP1 in the telomerase recruitment process. Here we develop an in vitro assay to quantitatively measure the contribution of the TEL-patch to telomerase recruitment--binding and extension of the first telomeric repeat. We also demonstrate that the TEL-patch contributes to the translocation step of the telomerase reaction. Finally, our quantitative observations indicate that the TEL-patch stabilizes the association between telomerase and telomeric DNA substrates, providing a molecular explanation for its contributions to telomerase recruitment and action.


How and when peers' positive mood influences employees' voice.
Employees often assess whether the social context is favorable for them to speak out, yet little research has investigated how the target's mood might influence the actor's voice behavior. From an affect-as-social-information perspective, we explored such potential effects of the target's mood on the actor's promotive voice in 2 empirical studies. In a scenario-based study with 142 MBA students (Study 1), the target's positive mood was positively associated with the actor's intentions to engage in promotive voice toward that target, mediated by the actor's perceived psychological safety. This mediated relationship was stronger when (a) the quality of the relationship between the actor and the target was poor or (b) the actor had a lower social status than the target. We replicated these results in Study 2, a correlational field study with 572 dyads nested within 142 members of 30 teams, where the actor's promotive voice behaviors (rather than intentions) were measured.


Diffusiophoresis in ionic surfactants: effect of micelle formation.
We explore the consequences of micelle formation for diffusiophoresis of charged colloidal particles in ionic surfactant concentration gradients, using a quasi-chemical association model for surfactant self assembly. The electrophoretic contribution to diffusiophoresis is determined by re-arranging the Nernst-Planck equations, and the chemiphoretic contribution is estimated by making plausible approximations for the density profiles in the electrical double layer surrounding the particle. For sub-micellar solutions we find that a particle will typically be propelled down the concentration gradient, although electrophoresis and chemiphoresis are finely balanced and the effect is sensitive to the detailed parameter choices and simplifying assumptions in the model. Above the critical micelle concentration (CMC), diffusiophoresis becomes much weaker and may even reverse sign, due to the fact that added surfactant goes into building micelles and not augmenting the monomer or counterion concentrations. We present detailed calculations for sodium dodecyl sulfate (SDS), finding that the typical drift speed for a colloidal particle in a ∼100 μm length scale SDS gradient is ∼1 μm s-1 below the CMC, falling to ⪅0.2 μm s-1 above the CMC. These predictions are broadly in agreement with recent experimental work.


Angular dependence of polarizing beam-splitter cubes.
The field-of-view dependence of polarizing beam-splitter cubes has been studied to characterize their behavior in imaging systems such as optical computers and optical correlators and in other applications that involve noncollimated light. Significant polarization aberration is present in polarizing beamsplitter cubes for two reasons: (1) the s- and p-component orientations, which define the polarizing axes, at the beam-splitting interface vary with the direction of propagation, and (2) the performance of the coating is a function of the angle of incidence. We describe the polarization aberration of a polarizing beam-splitter cube in terms of its diattenuation (polarizing efficiency). We use an imaging polarimeter to measure six figures of merit for three polarizing beam-splitter cubes demonstrating typical polarization aberrations. Finally, we derive the Mueller matrix for a polarizing beam-splitter cube in terms of the s and p transmittance and reflectance and the phase retardances, the parameters generally calculated with thin-film analysis programs.


Surgical options for patients with shoulder pain.
Shoulder pain is a common musculoskeletal complaint in the community, which can arise from diverse causes. Regardless of the cause, mild cases can often be effectively treated conservatively, with options including rest, physiotherapy, pain relief and glucocorticoid injections. If conservative strategies fail after a 3-6 month period then surgery might be considered. Generally, the proportion of patients with shoulder pain who require surgery is small. When surgery is considered, a clear diagnosis and structural information from imaging are required. The indications for surgery, and success rate, depend on the specific diagnosis as well as on the individual clinical presentation. Evidence from case series suggest that surgical interventions for shoulder pain are effective when used appropriately. This article outlines the surgical management of the most common painful conditions that affect the shoulder, including impingement, rotator cuff tear, frozen shoulder, osteoarthritis, rheumatoid arthritis and calcific tendonitis.


Effect of environmental changes on the ghosting of distant objects in twin-glazed windows.
Ghost images of distant objects are sometimes visible in twin-glazed window units. This is often caused by a pressure differential between the enclosed airspace and the surrounding atmosphere, which distorts the window. The ghost images result from internal reflections where the panes are not parallel. Even if the window is distortion-free for a particular atmospheric pressure and temperature, variations in either of these parameters will introduce distortion. Expressions are derived that relate the magnitude of the ghost separation to the structural parameters of the window and variations in atmospheric pressure and temperature of the enclosed airspace. Ghost separations of half a degree may often be expected as a result of environmental changes.


Evidence of pervasive biologically functional secondary structures within the genomes of eukaryotic single-stranded DNA viruses.
Single-stranded DNA (ssDNA) viruses have genomes that are potentially capable of forming complex secondary structures through Watson-Crick base pairing between their constituent nucleotides. A few of the structural elements formed by such base pairings are, in fact, known to have important functions during the replication of many ssDNA viruses. Unknown, however, are (i) whether numerous additional ssDNA virus genomic structural elements predicted to exist by computational DNA folding methods actually exist and (ii) whether those structures that do exist have any biological relevance. We therefore computationally inferred lists of the most evolutionarily conserved structures within a diverse selection of animal- and plant-infecting ssDNA viruses drawn from the families Circoviridae, Anelloviridae, Parvoviridae, Nanoviridae, and Geminiviridae and analyzed these for evidence of natural selection favoring the maintenance of these structures. While we find evidence that is consistent with purifying selection being stronger at nucleotide sites that are predicted to be base paired than at sites predicted to be unpaired, we also find strong associations between sites that are predicted to pair with one another and site pairs that are apparently coevolving in a complementary fashion. Collectively, these results indicate that natural selection actively preserves much of the pervasive secondary structure that is evident within eukaryote-infecting ssDNA virus genomes and, therefore, that much of this structure is biologically functional. Lastly, we provide examples of various highly conserved but completely uncharacterized structural elements that likely have important functions within some of the ssDNA virus genomes analyzed here.


Temporal order detection and coding in nervous systems.
This letter discusses temporal order coding and detection in nervous systems. Detection of temporal order in the external world is an adaptive function of nervous systems. In addition, coding based on the temporal order of signals can be used as an internal code. Such temporal order coding is a subset of temporal coding. We discuss two examples of processing the temporal order of external events: the auditory location detection system in birds and the visual direction detection system in flies. We then discuss how somatosensory stimulus intensities are translated into a temporal order code in the human peripheral nervous system. We next turn our attention to input order coding in the mammalian cortex. We review work demonstrating the capabilities of cortical neurons for detecting input order. We then discuss research refuting and demonstrating the representation of stimulus features in the cortex by means of input order. After some general theoretical considerations on input order detection and coding, we conclude by discussing the existing and potential use of input order coding in neuromorphic engineering.


Dual effects of cyclic ADP-ribose on sarcoplasmic reticulum Ca2+ release and storage in cardiac myocytes isolated from guinea-pig and rat ventricle.
The actions of cyclic ADP-ribose (cADPR), a regulator of Ca2+-induced Ca2+ release (CICR), were investigated on Ca2+ release and sarcoplasmic reticulum (SR) Ca2+ loading in cardiac myocytes at physiological temperature. In guinea-pig ventricular cells, cADPR, applied via patch pipette or from photorelease of its caged derivative, increased contraction amplitude and whole-cell Ca2+ transients, without affecting SR Ca2+ load (measured in response to rapid caffeine application). Under voltage-clamp conditions, photorelease of caged cADPR enhanced Ca2+ transient magnitude without affecting the peak amplitude of L-type Ca2+ current or its rate of decay, indicative of an increase in CICR gain. In rat permeabilised ventricular myocytes, rapid application of cADPR increased Ca2+ spark frequency within 30 s, and this effect was maintained over a 10 min exposure. Enhancement of spark frequency was not associated with changes in SR Ca2+ load at 30 s and 3 min of exposure to cADPR; however, prolonged exposure (10 min) was associated with an increased SR Ca2+ load (32+/-7%). The observations are consistent with dual actions of cADPR: a rapid effect on CICR that does not depend on an increased SR Ca2+ load, and an additional slower effect that is associated with enhanced SR Ca2+ levels.