Biodiversity responds to increasing climatic extremes in a biome-specific manner.
An unprecedented rate of global environmental change is predicted for the next century. The response to this change by ecosystems around the world is highly uncertain. To address this uncertainty, it is critical to understand the potential drivers and mechanisms of change in order to develop more reliable predictions. Australia's Long Term Ecological Research Network (LTERN) has brought together some of the longest running (10-60years) continuous environmental monitoring programs in the southern hemisphere. Here, we compare climatic variables recorded at five LTERN plot network sites during their period of operation and place them into the context of long-term climatic trends. Then, using our unique Australian long-term datasets (total 117 survey years across four biomes), we synthesize results from a series of case studies to test two hypotheses: 1) extreme weather events for each plot network have increased over the last decade, and; 2) trends in biodiversity will be associated with recent climate change, either directly or indirectly through climate-mediated disturbance (wildfire) responses. We examined the biodiversity responses to environmental change for evidence of non-linear behavior. In line with hypothesis 1), an increase in extreme climate events occurred within the last decade for each plot network. For hypothesis 2), climate, wildfire, or both were correlated with biodiversity responses at each plot network, but there was no evidence of non-linear change. However, the influence of climate or fire was context-specific. Biodiversity responded to recent climate change either directly or indirectly as a consequence of changes in fire regimes or climate-mediated fire responses. A national long-term monitoring framework allowed us to find contrasting species abundance or community responses to climate and disturbance across four of the major biomes of Australia, highlighting the need to establish and resource long-term monitoring programs across representative ecosystem types, which are likely to show context-specific responses.


Nutritional requirements of shigellae for growth in a minimal medium.
Most (about 81%) of the clinical isolates of shigellae that were tested failed to grow in a minimal medium. Of the auxotrophic isolates belonging to the four Shigella species, 98% grew in a minimal medium containing methionine, nicotinic acid, and tryptophan. The combination of methionine and tryptophan appears to be an obligatory requirement for Shigella dysenteriae serotype 1 strains, while the combination of nicotinic acid and tryptophan appears to be obligatory for serotype 2. Requirements which varied in other isolates were, however, genetically stable, indicating that the auxotypes may be useful as epidemiological markers. Cultures of shigellae in liquid minimal medium containing the above three supplements showed rapid growth and gave reasonably high cell yields.


Biophysical analysis of drug efficacy on C. elegans models for neurodegenerative and neuromuscular diseases.
Caenorhabditis elegans has emerged as a powerful model organism for drug screening due to its cellular simplicity, genetic amenability and homology to humans combined with its small size and low cost. Currently, high-throughput drug screening assays are mostly based on image-based phenotyping with the focus on morphological-descriptive traits not exploiting key locomotory parameters of this multicellular model with muscles such as its thrashing force, a critical biophysical parameter when screening drugs for muscle-related diseases. In this study, we demonstrated the use of a micropillar-based force assay chip in combination with a fluorescence assay to evaluate the efficacy of various drugs currently used in treatment of neurodegenerative and neuromuscular diseases. Using this two-dimensional approach, we showed that the force assay was generally more sensitive in measuring efficacy of drug treatment in Duchenne Muscular Dystrophy and Parkinson's Disease mutant worms as well as partly in Amyotrophic Lateral Sclerosis model. These results underline the potential of our force assay chip in screening of potential drug candidates for the treatment of neurodegenerative and neuromuscular diseases when combined with a fluorescence assay in a two-dimensional analysis approach.


The bioterrorism threat and dual-use biotechnological research: an Israeli perspective.
Israel has a long history of concern with chemical and biological threats, since several hostile states in the Middle East are likely to possess such weapons. The Twin-Tower terrorist attacks and Anthrax envelope scares of 2001 were a watershed for public perceptions of the threat of unconventional terror in general and of biological terror in particular. New advances in biotechnology will only increase the ability of terrorists to exploit the burgeoning availability of related information to develop ever-more destructive bioweapons. Many areas of modern biological research are unavoidably dual-use by nature. They thus have a great potential for both help and harm; and facilitating the former while preventing the latter remains a serious challenge to researchers and governments alike. This article addresses how Israel might best (1) prevent hostile elements from obtaining, from Israel's biological research system, materials, information and technologies that might facilitate their carrying out a biological attack, while (2) continuing to promote academic openness, excellence and other hallmarks of that system. This important and sensitive issue was assessed by a special national committee, and their recommendations are presented and discussed. One particularly innovative element is the restructuring and use of Israel's extensive biosafety system to also address biosecurity goals, with minimal disruption or delay.


The Association Between Facilitator Competent Adherence and Outcomes in Parenting Programs: a Systematic Review and SWiM Analysis.
There is increasing interest about the fidelity with which interventions are implemented because it is theorized that better implementation fidelity by facilitators is associated with better participant outcomes. However, in the parenting program literature, there is mixed evidence on the relationship between implementation fidelity and outcomes. This paper provides a synthesis of the evidence on the relationship between facilitator delivery and outcomes in the parenting program literature. Following PRISMA guidelines, this paper synthesizes the results of a systematic review of studies on parenting programs aiming to reduce violence against children and child behavior problems. Specifically, it examines associations between observational measures of facilitator competent adherence and parent and child outcomes. A meta-analysis was not feasible due to study heterogeneity. As a result, Synthesis Without Meta-Analysis guidelines were followed. Searches in electronic databases, reference searching, forward citation tracking, and expert input identified 9653 articles. After screening using pre-specified criteria, 18 articles were included. The review found that most studies (n = 13) reported a statistically significant positive relationship with at least one parent or child outcome. However, eight studies reported inconsistent findings across outcomes, and four studies found no association with outcomes. The results suggest that better facilitator competent adherence is generally associated with positive parent and child outcomes. However, this finding is weakened by the methodological heterogeneity of included studies and due to the wide variety of ways in which studies conceptualized competent adherence-outcome relationships.


Pegfilgrastim use and bone pain: a cohort study of community-based cancer patients.
Bone pain is a common adverse effect of the granulocyte colony-stimulating factors filgrastim and pegfilgrastim. However, the incidence of reported bone pain varies and therapies to mitigate this adverse effect are limited to case reports and one randomized controlled trial. The purpose of this study was to describe pegfilgrastim use, the incidence and treatment of bone pain, and rate of severe or febrile neutropenia among cancer patients receiving pegfilgrastim at a metropolitan, hospital-based, community cancer center. This retrospective chart review included the first 100 adult oncology patients who received at least one dose of pegfilgrastim from 1 January 2012 to 31 December 2012. Descriptive analyses were used to evaluate the primary and secondary outcomes. Of the identified cases, 69 cancer patients were evaluable. Most patients (74%) received pegfilgrastim for primary prophylaxis. Pegfilgrastim-associated bone pain occurred in 19% and loratadine was the most common medication used to treat it. Among the patients who received pegfilgrastim for primary prophylaxis, 8% were hospitalized for febrile neutropenia. Among those hospitalized for febrile neutropenia, 64% had not received pegfilgrastim for primary prophylaxis. Pegfilgrastim is commonly used for primary prophylaxis during the first cycle of chemotherapy. Hospitalizations for febrile neutropenia occurred most commonly among patients without primary prophylaxis. Pegfilgrastim-associated bone pain occurred in a similar percentage, as reported in randomized controlled trials but less than that reported by survey. Loratadine was the most commonly employed medication to mitigate this adverse effect.


Premature senescence of T-cell subsets in axial spondyloarthritis.
To investigate the possible occurrence of early thymic failure and premature senescence of naïve and memory T-cells in patients with axial spondyloarthritis (aSpA). Prospective, cross-sectional study of consecutive patients with aSpA (n=51), rheumatoid arthritis (RA, n=51) and healthy controls (HCs, n=50). Demographic, clinical and laboratory parameters were collected in all patients and we isolated naïve (CD45RA(+)) and memory (CD45RO(+)) CD4(+) and CD8(+) T-cell subsets by MACS technology. T-cell receptor rearrangement excision circle (TREC) and telomere length were measured by real-time PCR. We used TRECs as a surrogate for thymus function and telomere length as an indicator of cellular senescence. Telomerase activity was analysed with the Telomeric Repeat Amplification Protocols. We observed a premature decline of thymic output in patients with aSpA and patients with RA compared with HCs as indicated by a reduction of TREC levels in naive T-cells (aSpA: age adjusted regression coefficient (regcoeff) for CD4(+)CD45RA(+) T-cells -2.566, p=0.023; RA regcoeff=-2.844, p=0.008). Telomere length of all CD4(+) and CD8(+) T-cell subsets was reduced in young patients with aSpA compared with HCs, whereas data for patients with RA were comparable with HCs. Telomerase activity was inversely correlated with telomere length in HCs (correlation coefficient (corcoeff)=-0.532, p<0.001) but not in patients with aSpA (corcoeff=-0.056, p=0.697) and RA (corcoeff=-0.003, p=0.982). Our data indicate an age-inappropriate shrinkage of thymic output, an inappropriate shortening of telomeres in young patients with aSpA and an impaired telomerase enzyme in patients with aSpA and RA.


Lower leg arterial calcification assessed by high-resolution peripheral quantitative computed tomography is associated with bone microstructure abnormalities in women.
In older women, the presence of lower leg arterial calcification assessed by high-resolution peripheral quantitative computed tomography is associated with relevant bone microstructure abnormalities at the distal tibia and distal radius. Here, we report the relationships of bone geometry, volumetric bone mineral density (BMD) and bone microarchitecture with lower leg arterial calcification (LLAC) as assessed by high-resolution peripheral quantitative computed tomography (HR-pQCT). We utilized the Hertfordshire Cohort Study (HCS), where we were able to study associations between measures obtained from HR-pQCT of the distal radius and distal tibia in 341 participants with or without LLAC. Statistical analyses were performed separately for women and men. We used linear regression models to investigate the cross-sectional relationships between LLAC and bone parameters. The mean (SD) age of participants was 76.4 (2.6) and 76.1 (2.5) years in women and men, respectively. One hundred and eleven of 341 participants (32.6 %) had LLAC that were visible and quantifiable by HR-pQCT. The prevalence of LLAC was higher in men than in women (46.4 % (n = 83) vs. 17.3 % (n = 28), p < 0.001). After adjustment for confounding factors, we found that women with LLAC had substantially lower Ct.area (β = -0.33, p = 0.016), lower Tb.N (β = -0.54, p = 0.013) and higher Tb.Sp (β = 0.54, p = 0.012) at the distal tibia and lower Tb.Th (β = -0.49, p = 0.027) at the distal radius compared with participants without LLAC. Distal radial or tibial bone parameter analyses in men according to their LLAC status revealed no significant differences with the exception of Tb.N (β = 0.27, p = 0.035) at the distal tibia. In the HCS, the presence of LLAC assessed by HR-pQCT was associated with relevant bone microstructure abnormalities in women. These findings need to be replicated and further research should study possible pathophysiological links between vascular calcification and osteoporosis.


Making Sense of an Unknown Terrain: How Parents Understand Self-Harm in Young People.
Self-harm is common in young people, and can have profound effects on parents and other family members. We conducted narrative interviews with 41 parents and other family members of 38 young people, aged up to 25, who had self-harmed. Most of the participants were parents but included one sibling and one spouse. This article reports experiences of the parent participants. A cross-case thematic analysis showed that most participants were bewildered by self-harm. The disruption to their worldview brought about by self-harm prompted many to undergo a process of "sense-making"-by ruminative introspection, looking for information, and building a new way of seeing-to understand and come to terms with self-harm. Most participants appeared to have been successful in making sense of self-harm, though not without considerable effort and emotional struggle. Our findings provide grounds for a deeper socio-cultural understanding of the impact of self-harm on parents.


When thoughts become action: an fMRI paradigm to study volitional brain activity in non-communicative brain injured patients.
The assessment of voluntary behavior in non-communicative brain injured patients is often challenging due to the existence of profound motor impairment. In the absence of a full understanding of the neural correlates of consciousness, even a normal activation in response to passive sensory stimulation cannot be considered as proof of the presence of awareness in these patients. In contrast, predicted activation in response to the instruction to perform a mental imagery task would provide evidence of voluntary task-dependent brain activity, and hence of consciousness, in non-communicative patients. However, no data yet exist to indicate which imagery instructions would yield reliable single subject activation. The aim of the present study was to establish such a paradigm in healthy volunteers. Two exploratory experiments evaluated the reproducibility of individual brain activation elicited by four distinct mental imagery tasks. The two most robust mental imagery tasks were found to be spatial navigation and motor imagery. In a third experiment, where these two tasks were directly compared, differentiation of each task from one another and from rest periods was assessed blindly using a priori criteria and was correct for every volunteer. The spatial navigation and motor imagery tasks described here permit the identification of volitional brain activation at the single subject level, without a motor response. Volunteer as well as patient data [Owen, A.M., Coleman, M.R., Boly, M., Davis, M.H., Laureys, S., Pickard J.D., 2006. Detecting awareness in the vegetative state. Science 313, 1402] strongly suggest that this paradigm may provide a method for assessing the presence of volitional brain activity, and thus of consciousness, in non-communicative brain-injured patients.


Biogeochemistry of hypersaline springs supporting a mid-continent marine ecosystem: an analogue for martian springs?
Hypersaline springs that host unique mid-continent marine ecosystems were examined in central Manitoba, Canada. The springs originate from a reflux of glacial meltwater that intrudes into underlying bedrock and dissolved buried salt beds. Two spring types were distinguished based both on flow rate and geochemistry. High flow springs (greater than 10 L/s) hosted extensive marine microbial mats, which were dominated by algae but also included diverse microbes. These varied somewhat between springs as indicated by changes in profiles of fatty acid methyl esters. Culture studies confirmed the presence of sulfate-reducing bacteria in sediments at the high flow sites. In contrast, low flow springs were affected by solar evaporation, increasing salinity, and temperature. These low flow springs behaved more like closed nutrient-limited systems and did not support microbial mats. Direct comparison of the high and low flow springs revealed interesting implications for the potential to record biosignatures in the rock record. High flow springs have abundant, well-developed microbial mats, which desiccate and are cemented along the edges of the spring pools; however, the high mass flux overwhelms any geochemical signature of microbial activity. In contrast, the nutrient-limited low flow sites develop strong geochemical signatures of sulfate reduction, even in the absence of microbial mats, due to less dilution with the lower flows. Geochemical and physical evidence for life did not correlate with the abundance of microbial life but, rather, with the extent to which the biological system formed a closed ecosystem.


Mobility of antimicrobial resistance across serovars and disease presentations in non-typhoidal Salmonella from animals and humans in Vietnam.
Non-typhoidal Salmonella (NTS) is a major cause of bacterial enterocolitis globally but also causes invasive bloodstream infections. Antimicrobial resistance (AMR) hampers the treatment of these infections and understanding how AMR spreads between NTS may help in developing effective strategies. We investigated NTS isolates associated with invasive disease, diarrhoeal disease and asymptomatic carriage in animals and humans from Vietnam. Isolates included multiple serovars and both common and rare phenotypic AMR profiles; long- and short-read sequencing was used to investigate the genetic mechanisms and genomic backgrounds associated with phenotypic AMR profiles. We demonstrate concordance between most AMR genotypes and phenotypes but identified large genotypic diversity in clinically relevant phenotypes and the high mobility potential of AMR genes (ARGs) in this setting. We found that 84 % of ARGs identified were located on plasmids, most commonly those containing IncHI1A_1 and IncHI1B(R27)_1_R27 replicons (33%), and those containing IncHI2_1 and IncHI2A_1 replicons (31%). The vast majority (95%) of ARGS were found within 10 kbp of IS6/IS26 elements, which provide plasmids with a mechanism to exchange ARGs between plasmids and other parts of the genome. Whole genome sequencing with targeted long-read sequencing applied in a One Health context identified a comparatively limited number of insertion sequences and plasmid replicons associated with AMR. Therefore, in the context of NTS from Vietnam and likely for other settings as well, the mechanisms by which ARGs move contribute to a more successful AMR profile than the specific ARGs, facilitating the adaptation of bacteria to different environments or selection pressures.


Isolation, identification and changes in the composition of lactic acid bacteria during the malting of two different barley cultivars.
Malt has a complex microbial population, which changes as the malting process commences. Little is known about the proliferation of lactic acid bacteria (LAB) in each of the malting phases. In this study, we determined the number of LAB present in the different phases of malting with Clipper and Prisma barley cultivars. The strains were identified to species level by using numerical analysis of total soluble cell protein patterns, RAPD-PCR banding patterns and 16S rRNA sequencing. The number of viable LAB in the barley before steep was higher in Prisma than Clipper (7.6 x 10(4) and 1.2 x 10(3) cfu/g, respectively). Despite this, the number of viable cells recorded in the first steep water was slightly higher for Clipper (9.0 x 10(5) cfu/g) than Prisma (5.5 x 10(5) cfu/g). More or less, the same cell numbers were recorded for the two barley cultivars after the first and second dry stands. Both cultivars displayed more or less the same cell numbers (3.7 x 10(7) for Clipper and 3.2 x 10(7) cfu/g for Prisma) after the third day of germination. However, a higher number of LAB were detected in the kilned Prisma malt (6.9 x 10(4) cfu/g) than the Clipper malt (1.5 x 10(4) cfu/g). Leuconostoc argentinum, Le. lactis and Weissella confusa were the most predominant in both cultivars. A few strains were identified as W. paramesenteroides (four strains), Lactobacillus casei (five strains), Lactococcus lactis (five strains) and Lb. rhamnosus (two strains). Lb. casei and Lb. rhamnosus were not isolated from the Prisma cultivar, while W. paramesenteroides and Lc. lactis were absent in the Clipper cultivar. Kilned malt of the Clipper cultivar contained predominantly Le. argentinum, whereas the Prisma cultivar contained mainly Lc. lactis. To our knowledge. this is the first report of LAB in Clipper and Prisma barley and the various malting phases. The influence of the various groups of lactic acid bacteria on the fermenting ability of brewers' yeast is currently being determined.


The reconstitution of functional respiratory chains in membranes from electron-transport-deficient mutants of Escherichia coli as demonstrated by quenching of atebrin fluorescence.
Measurements were made of electron-transport-dependent quenching of atebrin fluorescence in particles prepared from mutants of Escherichia coli unable to synthesize either ubiquinone or haem. Such quenching was either absent (in haem-deficient particles) or decreased (in ubiquinone-deficient particles), but was restored under conditions used to reconstitute oxidase activities. It is concluded that the reconstitution of oxidase activity, in both cases, is associated with the formation of a functional, proton-translocating, respiratory chain.


Simulating Crop Evapotranspiration Response under Different Planting Scenarios by Modified SWAT Model in an Irrigation District, Northwest China.
Modelling crop evapotranspiration (ET) response to different planting scenarios in an irrigation district plays a significant role in optimizing crop planting patterns, resolving agricultural water scarcity and facilitating the sustainable use of water resources. In this study, the SWAT model was improved by transforming the evapotranspiration module. Then, the improved model was applied in Qingyuan Irrigation District of northwest China as a case study. Land use, soil, meteorology, irrigation scheduling and crop coefficient were considered as input data, and the irrigation district was divided into subdivisions based on the DEM and local canal systems. On the basis of model calibration and verification, the improved model showed better simulation efficiency than did the original model. Therefore, the improved model was used to simulate the crop evapotranspiration response under different planting scenarios in the irrigation district. Results indicated that crop evapotranspiration decreased by 2.94% and 6.01% under the scenarios of reducing the planting proportion of spring wheat (scenario 1) and summer maize (scenario 2) by keeping the total cultivated area unchanged. However, the total net output values presented an opposite trend under different scenarios. The values decreased by 3.28% under scenario 1, while it increased by 7.79% under scenario 2, compared with the current situation. This study presents a novel method to estimate crop evapotranspiration response under different planting scenarios using the SWAT model, and makes recommendations for strategic agricultural water management planning for the rational utilization of water resources and development of local economy by studying the impact of planting scenario changes on crop evapotranspiration and output values in the irrigation district of northwest China.


Phenotype-genotype relationships in monogenic disease: lessons from the thalassaemias.
The remarkable phenotypic diversity of the beta-thalassaemias reflects the heterogeneity of mutations at the beta-globin locus, the action of many secondary and tertiary modifiers, and a wide range of environmental factors. It is likely that phenotype-genotype relationships will be equally complex in the case of many monogenic diseases. These findings highlight the problems that might be encountered in defining the relationship between the genome and the environment in multifactorial disorders, in which the degree of heritability might be relatively low and several environmental agents are involved. They also emphasize the value of an understanding of phenotype-genotype relationships in designing approaches to gene therapy.


Individual and organizational risk factors of work-related violence in the Danish elder care.
The aim of this study was to explore individual and organizational risk factors of exposure to violence in the Elder Care Sector. A questionnaire study conducted among 8134 health-care workers in the elder care in 36 municipalities was carried out between October 2004 and May 2005. Participation in the study was voluntary and data were treated confidentially. Individual factors such as low age and seniority seemed to increase the risk of violence. Type of institution (nursing homes), occupation (health care assistants), shift-work, evening or night work, frequent physical contact with clients, higher emotional demands, a lower level of involvement at the workplace, lower leadership quality, and more role conflicts increased the risk of work-related violence. On the other hand, the study also demonstrated that the least violence was found at workplaces with high leadership quality, no role conflicts and high involvement in the workplace. Study strengths and limitations: The strengths of the study are that it is a large representative study consisting of Danish elder care workers in 36 municipalities and has a high response rate. A limitation is that the study is cross-sectional making it impossible to decide the direction of causal relationships. The results of the study indicate that there is a possibility of reducing the prevalence of violence towards care-workers in the elder care if policies and resources to improve the working environment are implemented.


Pronounced transcriptional regulation of apoptotic and TNF-NF-kappa-B signaling genes during the course of thymoquinone mediated apoptosis in HeLa cells.
Thymoquinone (TQ) is the active ingredient extracted from the essential oil of Nigella sativa. A number of studies implicated TQ as an antitumor agent. In this study, cytotoxic effects of the oil of N. sativa and TQ were evaluated on human cervical cancer cell line, HeLa cells. IC50 value was ~0.125 μl/ml for N. sativa oil preparations and 12.5 μM for TQ. TQ strongly inhibited wound healing at all concentrations ranging from 12.5 to 100 μM in a scratch wound healing assay. Additionally, induction of apoptosis by TQ was assessed by Giemsa staining and TQ was found to induce apoptosis in cancer cells especially at concentrations of 50 and 100 μM. TQ-mediated transcriptional regulation of 84 genes involved in apoptosis was studied using a PCR array. At low dose (12.5 μM), TQ was found to induce expression of four pro-apoptotic genes: BIK (~22.7-fold), FASL (~2.9-fold), BCL2L10 (~2.1-fold), and CASP1 (~2-fold). TQ was also found to reduce the expression of an anti-apoptotic gene implicated in NF-kappa-B signaling and cancer: RELA (~8-fold). At high dose (100 μM), TQ mediated the expression of 21 genes implicated directly in apoptosis (6 genes), TNF signaling (10 genes), and NF-kappa-B signaling (3 genes) such as BIK, BID, TNFRSF10A, TNFRSF10B, TNF, TRAF3, RELA, and RELB. In conclusion, this study implicates the role of TQ in the inhibition of cancer cell proliferation and migration. At the same time, our results strongly suggest that TQ intervenes with TNF and NF-kappa-B signaling during TQ-mediated induction of apoptosis in cancer cells.


Sequential Therapeutic Response Modeling for Tumor Treatment Using Computational Hybrid Control Systems Approach.
Tumorigenesis is due to uncontrolled cell division arising from mutations and alterations in the proliferative controls of the cell population. The fight against tumor growth and development has often relied on combination therapy that has been acclaimed as one of the main standards of care in cancer therapeutics and prevention of drug-related resistances. The toxicity of the combinatorial drugs raises a significant concern whenever patients take two or more drugs concurrently at the maximum tolerated dose. A promising solution in tumor treatment involves the administration of the drugs in an alternating or sequential fashion rather than a simultaneous manner. In this paper, we investigate how feasible such an approach is from a mathematical perspective and propose a switched hybrid control systems framework. We explore the response of tumor cells dynamics to sequential drugs administration with the aid of a time-dependent switching strategy. A transit compartmentalized model is employed to describe the tumor cells progression to death. The design of the time-based drug switching logic ensures the proliferating tumor cells are repressed. Simulation results are provided using the tumor growth dynamics with sequential drugs intake to demonstrate the effectiveness of the proposed method in reducing the tumor size. This paper is the first attempt to provide a switched hybrid control systems framework on sequential drug administration to biomedical researchers and clinicians.


21-Hydroxy-6,19-epoxyprogesterone: A Promising Therapeutic Agent and a Molecular Tool for Deciphering Glucocorticoid Action.
Glucocorticoids are steroid hormones that exert most of their effects through their binding to the glucocorticoid receptor (GR), a ligand regulated transcription factor. Although glucocorticoids are widely used in the clinic, their usage in chronic therapies provokes severe adverse reactions. In the quest for safer glucocorticoids a dissociated model was established that proposes a disconnection between GR activated pathways responsible of desired pharmacological effects and pathways involved in adverse GR reactions. Under this model, a myriad of steroidal and non-steroidal compounds has been characterized, with most of them still producing side effects. X-ray crystallographic studies followed by molecular dynamics analysis led research to insights on the receptor Ligand Binding Domain (LBD), which undergoes specific ligand dependent conformational changes that influence receptor activities. In this sense, the flexibility of the ligand structure would contribute to the final GR outcome. Here, we review different data of 21-hydroxy-6,19-epoxyprogesterone (21OH-6,19OP), a rigid steroid with potential pharmaceutical interest due to its anti-inflammatory and immunosuppressive activities, lacking several GR adverse reactions. The rigid structure endows this compound with an enhanced selectivity towards GR. Molecular characterization of the GR/21OH-6,19OP complex revealed specific intermediate conformations adopted by the receptor that would explain the influence on GR dimerization and the recruitment of a specific set of GR transcription modulators. We summarize recent data that will contribute to understand the complexity of glucocorticoid response.


Impact of monocarboxylate transporter-8 deficiency on the hypothalamus-pituitary-thyroid axis in mice.
In patients, inactivating mutations in the gene encoding the thyroid hormone-transporting monocarboxylate transporter 8 (Mct8) are associated with severe mental and neurological deficits and disturbed thyroid hormone levels. The latter phenotype characterized by high T3 and low T4 serum concentrations is replicated in Mct8 knockout (ko) mice, indicating that MCT8 deficiency interferes with thyroid hormone production and/or metabolism. Our studies of Mct8 ko mice indeed revealed increased thyroidal T3 and T4 concentrations without overt signs of a hyperactive thyroid gland. However, upon TSH stimulation Mct8 ko mice showed decreased T4 and increased T3 secretion compared with wild-type littermates. Moreover, similar changes in the thyroid hormone secretion pattern were observed in Mct8/Trhr1 double-ko mice, which are characterized by normal serum T3 levels and normal hepatic and renal D1 expression in the presence of very low T4 serum concentrations. These data strongly indicate that absence of Mct8 in the thyroid gland affects thyroid hormone efflux by shifting the ratio of the secreted hormones toward T3. To test this hypothesis, we generated Mct8/Pax8 double-mutant mice, which in addition to Mct8 lack a functional thyroid gland and are therefore completely athyroid. Following the injection of these animals with either T4 or T3, serum analysis revealed T3 concentrations similar to those observed in Pax8 ko mice under thyroid hormone replacement, indicating that indeed increased thyroidal T3 secretion in Mct8 ko mice represents an important pathogenic mechanism leading to the high serum T3 levels.


Search for Higgs bosons predicted in two-Higgs-doublet models via decays to tau lepton pairs in 1.96 TeV pp collisions.
We present the results of a search for Higgs bosons predicted in two-Higgs-doublet models, in the case where the Higgs bosons decay to tau lepton pairs, using 1.8 fb(-1) of integrated luminosity of pp collisions recorded by the CDF II experiment at the Fermilab Tevatron. Studying the mass distribution in events where one or both tau leptons decay leptonically, no evidence for a Higgs boson signal is observed. The result is used to infer exclusion limits in the two-dimensional space of tanbeta versus m(A) (the ratio of the vacuum expectation values of the two Higgs doublets and the mass of the pseudoscalar boson, respectively).


Firearm injury prevention counseling: a study of pediatricians' beliefs and practices.
Members of the Maryland Chapter of the American Academy of Pediatrics completed a mail survey on their beliefs and counseling practices related to firearm injury prevention. Respondents were skeptical of the protective value of firearms in the home and most were supportive of gun control measures. Only a fifth believed that most families with handguns keep them inaccessible to children; however, many seemed to believe that the children at risk were in practices other than their own. Among those providing direct ambulatory care, 40% had had a patient who had been shot. Seventy-four percent believed pediatricians have a responsibility to counsel families about firearms. Only 13% believed parents would be offended if guns were included in anticipatory guidance discussions. Even so, only 30% had ever provided such counseling. Just half of the respondents agreed that they knew what to tell families about firearms. Ninety percent were very likely to counsel parents to store guns unloaded and locked up, whereas 54% were very likely to advise parents to remove guns from the home. More than two thirds believed parents would heed their advice about storing firearms, and 30% believed parents would follow advice about having guns. Results suggest pediatricians are ready to counsel about firearm injury prevention, but are not yet doing so.


Temporal macrodynamics and microdynamics of the postoperative impedance at the tissue-electrode interface in deep brain stimulation patients.
To study the temporal dynamics of tissue impedance after deep brain stimulation (DBS). DBS therapy commonly employs a constant voltage approach, and current delivery to the tissue is a function of electrode-tissue impedance. It is presumed that impedance fluctuates early postimplantation, with implications for variations in current delivery and therapeutic efficacy. We hypothesised that the largest variation will be recorded early after surgery, followed by stabilisation. Review of impedance checks of implanted DBS systems at standard parameters during the first five months postimplantation. All measurement time points were binned into 1-week periods, and we used repeated measures analysis of variance with Tukey pairwise multiple comparisons correction. The analysis was repeated after normalising impedance values for each subject to that patient's baseline value. There was an initial (non-significant) drop in impedance at week 1, followed by significant increase at week 3 (p=0.0002). There were no further significant differences in impedance values at subsequent time points. Analysis of normalised data showed a significant difference between the initial measurement in postoperative week 1 (normalised value 1) and week 3 (normalised value 1.73, p<0.0001), with no further difference among the subsequent weekly points during the 5-month follow-up. No significant hourly variations were found at any time points. We found major changes in impedance within the first month postimplantation, with no further variation. This is an important confirmation in patients of this temporal dynamics of the impedance of implanted DBS hardware, with potential therapeutic implications.


Tissue-specific transcriptome assemblies of the marine medaka Oryzias melastigma and comparative analysis with the freshwater medaka Oryzias latipes.
The marine medaka Oryzias melastigma has been demonstrated as a novel model for marine ecotoxicological studies. However, the lack of genome and transcriptome reference has largely restricted the use of O. melastigma in the assessment of in vivo molecular responses to environmental stresses and the analysis of biological toxicity in the marine environment. Although O. melastigma is believed to be phylogenetically closely related to Oryzias latipes, the divergence between these two species is still largely unknown. Using Illumina high-throughput RNA sequencing followed by de novo assembly and comprehensive gene annotation, we provided transcriptomic resources for the brain, liver, ovary and testis of O. melastigma. We also investigated the possible extent of divergence between O. melastigma and O. latipes at the transcriptome level. More than 14,000 transcripts across brain, liver, ovary and testis in marine medaka were annotated, of which 5880 transcripts were orthologous between O. melastigma and O. latipes. Tissue-enriched genes were identified in O. melastigma, and Gene Ontology analysis demonstrated the functional specificity of the annotated genes in respective tissue. Lastly, the identification of marine medaka-enriched transcripts suggested the necessity of generating transcriptome dataset of O. melastigma. Orthologous transcripts between O. melastigma and O. latipes, tissue-enriched genes and O. melastigma-enriched transcripts were identified. Genome-wide expression studies of marine medaka require an assembled transcriptome, and this sequencing effort has generated a valuable resource of coding DNA for a non-model species. This transcriptome resource will aid future studies assessing in vivo molecular responses to environmental stresses and those analyzing biological toxicity in the marine environment.


Competency based training in robotic surgery: benchmark scores for virtual reality robotic simulation.
To develop benchmark scores of competency for use within a competency based virtual reality (VR) robotic training curriculum. This longitudinal, observational study analysed results from nine European Association of Urology hands-on-training courses in VR simulation. In all, 223 participants ranging from novice to expert robotic surgeons completed 1565 exercises. Competency was set at 75% of the mean expert score. Benchmark scores for all general performance metrics generated by the simulator were calculated. Assessment exercises were selected by expert consensus and through learning-curve analysis. Three basic skill and two advanced skill exercises were identified. Benchmark scores based on expert performance offered viable targets for novice and intermediate trainees in robotic surgery. Novice participants met the competency standards for most basic skill exercises; however, advanced exercises were significantly more challenging. Intermediate participants performed better across the seven metrics but still did not achieve the benchmark standard in the more difficult exercises. Benchmark scores derived from expert performances offer relevant and challenging scores for trainees to achieve during VR simulation training. Objective feedback allows both participants and trainers to monitor educational progress and ensures that training remains effective. Furthermore, the well-defined goals set through benchmarking offer clear targets for trainees and enable training to move to a more efficient competency based curriculum.


Prevalence of angina in women versus men: a systematic review and meta-analysis of international variations across 31 countries.
In the absence of previous international comparisons, we sought to systematically evaluate, across time and participant age, the sex ratio in angina prevalence in countries that differ widely in the rate of mortality due to myocardial infarction. We searched MEDLINE and EMBASE until February 2006 for healthy population studies published in any language that reported the prevalence of angina (Rose questionnaire) in women and men. We obtained myocardial infarction mortality rates from the World Health Organization. A total of 74 reports of 13,331 angina cases in women and 11,511 cases in men from 31 countries were included. Angina prevalence varied widely across populations, from 0.73% to 14.4% (population weighted mean 6.7%) in women and from 0.76% to 15.1% (population weighted mean 5.7%) in men, and was strongly correlated within populations between the sexes (r=0.80, P<0.0001). Angina prevalence showed a small female excess with a pooled random-effects sex ratio of 1.20 (95% CI 1.14 to 1.28, P<0.0001). This female excess was found across countries with widely differing myocardial infarction mortality rates in women (interquartile range 12.7 to 126.5 per 100,000), was particularly high in the American studies (1.40, 95% CI 1.28 to 1.52), and was higher among nonwhite ethnic groups than among whites. This sex ratio did not differ significantly by participant's age, the year the survey began, or the sex ratio for mortality due to myocardial infarction. Over time and at different ages, independent of diagnostic and treatment practices, women have a similar or slightly higher prevalence of angina than men across countries with widely differing myocardial infarction mortality rates.


Spatial and temporal heterogeneity of bacteria across an intertidal shellfish bed: implications for regulatory monitoring of faecal indicator organisms.
Routine bacterial monitoring of shellfish beds using indicator species is a common global practice designed to prevent human consumption of contaminated shellfish products. However, current bacteriological monitoring procedures which focus on the quantification of faecal indicator organisms (FIOs) as a proxy for microbial pollution may not be representative of total bacterial contamination levels present in shellfish harvesting areas. The objective of this study was to critically assess the accuracy of current monitoring strategies by quantifying the spatial (lateral and longitudinal distance) and temporal (seasonality and tidal state) concentrations of FIOs (Escherichia coli and total coliforms) within a single intertidal commercially harvested shellfish bed. Spatial and temporal FIO dynamics, including the effects of tidal state and seasonality, were quantified in mussel flesh and sediment samples from a single intertidal mussel (Mytilus edulis) bed. Our results confirmed that FIO concentrations across a shellfish bed were heterogeneous over larger spatial and temporal scales, but showed no relation to the concentrations of autochthonous bacteria, such as Vibrio spp., or the physico-chemical parameters of the sediment. These results have important implications for both public health and the economic prosperity of the shellfish industry, and demonstrate the importance of accommodating both spatial and temporal fluctuations in routine bacteriological monitoring protocols. We conclude that current FIO monitoring procedures may not accurately represent levels of microbial contamination within shellfish harvesting areas and that more robust microbiological testing procedures need developing.


Farm machinery injuries: the 15-year experience at an urban joint trauma center system in a rural state.
Farm machinery is a major source of injury. The objective of this study is to characterize the incidence, injury characteristics, and outcomes of patients admitted with farm machinery injuries (FMIs) to an urban joint trauma system in a rural state. A retrospective 15-year review of the trauma registries of the two trauma centers that function as a single state-designated Level I joint trauma center system was conducted. There were 65 admissions for FMIs at hospital A and 41 at hospital B; this represents under 0.4% of total trauma admissions. The patients ranged in age from 2 to 87 years. At hospital A, 89% of admitted patients sustained extremity injuries, 16% sustained torso trauma, 92% required surgical intervention, and the mortality rate was 0%. At hospital B, 60% of admitted patients sustained extremity injuries, 36.6% of patients sustained torso trauma, 63% required surgical intervention, and the mortality rate was 14.6%. Tractor-related injuries were responsible for 17% of admissions at hospital A and 69% at hospital B. Of the six fatalities, five were tractor related. The data demonstrate that FMIs affect people in nearly all decades of life. FMIs at the two hospitals had differing injury characteristics and outcomes, in large part secondary to the differing frequency of tractor-related injuries. FMIs frequently required surgical intervention.


Perceived Costs and Benefits of Early Childbearing: New Dimensions and Predictive Power.
Understanding the causes of early childbearing is important for reducing the persistently high rates of early births in the United States. Perceptions of possible benefits may contribute to these rates, while high opportunity costs may dissuade women from early childbearing. Perceptions of costs and benefits of pregnancy, as well as later experiences of pregnancy, were assessed for 701 nulligravid women aged 18-22 who entered the Relationship Dynamics and Social Life study in 2008-2009 and were interviewed weekly for up to 30 months. Bivariate t tests, chi-square tests and multivariable discrete-time event history analyses were used to assess associations of perceived personal consequences of childbearing (e.g., predicted financial costs), goals in potentially competing domains (opportunity costs) and social norms with subsequent pregnancy. Twenty percent of women reported that early childbearing would have more positive than negative personal consequences. Compared with other women, those who had a pregnancy during follow-up had, at baseline, more positive perceptions of the personal consequences of pregnancy and of their friends' approval of pregnancy, and greater desire for consumer goods. In multivariable analyses, only the scales assessing perceived personal consequences of childbearing and friends' approval of childbearing were associated with pregnancy (odds ratios, 2.0 and 1.2, respectively). Goals in potentially competing domains were not associated with pregnancy. Young women's perceptions of consequences of early childbearing predict subsequent pregnancy. That these perceptions are distinct from childbearing desires and from other dimensions of costs and benefits illustrates the complex attitudinal underpinnings of reproductive behavior.


Glucocorticoid receptor gene polymorphisms and disease activity during pregnancy and the postpartum period in rheumatoid arthritis.
The mechanism underlying the spontaneous improvement of rheumatoid arthritis (RA) during pregnancy and the subsequent postpartum flare is incompletely understood, and the disease course varies widely between pregnant RA patients. In pregnancy, total and free levels of cortisol increase gradually, followed by a postpartum decrease to prepregnancy values. The glucocorticoid receptor (GR) polymorphisms BclI and N363S are associated with relatively increased glucocorticoid (GC) sensitivity, whereas the 9β and ER22/23EK polymorphisms of the GR gene are associated with a relatively decreased GC sensitivity. We examined the relation between the presence of these GR polymorphisms and level of disease activity and disease course of RA during pregnancy and postpartum. We studied 147 participants of the PARA study (Pregnancy-Induced Amelioration of Rheumatoid Arthritis study), a prospective study investigating the natural improvement during pregnancy and the postpartum flare in women with RA. Patients were visited, preferably before pregnancy, at each trimester and at three postpartum time points. On all occasions, disease activity was scored by using DAS28. All patients were genotyped for the GR polymorphisms BclI, N363S, 9β, and ER22/23EK and divided in groups harboring either polymorphisms conferring increased GC sensitivity (BclI and N363S; GC-S patients) or polymorphisms conferring decreased GC sensitivity (9β or 9β + ER22/23EK; GC-I patients). Data were analyzed by using a mixed linear model, comparing GC-S patients with GC-I patients with respect to improvement during pregnancy and the postpartum flare. The cumulative disease activity was calculated by using time-integrated values (area under the curve, AUC) of DAS28 in GC-I patients versus GC-S patients. Separate analyses were performed according to the state of GC use. GC-S patients treated with GC had a significantly lower AUC of DAS28 in the postpartum period than did GC-I patients. This difference was not observed in patients who were not treated with GCs. During pregnancy, GC-S and GC-I patients had comparable levels of disease activity and course of disease. Differences in relative GC sensitivity, as determined by GR polymorphisms, are associated with the level of disease activity in the postpartum period in GC-treated patients, but they do not seem to influence the course of the disease per se.


Hepatic hyperplasia in noncirrhotic fatty livers: is obesity-related hepatic steatosis a premalignant condition?
It is not known whether obesity increases the risk for hepatocellular carcinoma (HCC) simply because it promotes cirrhosis, a general risk factor for HCC, or via some other mechanism that operates independently of cirrhosis. If the latter occurs, then hepatocyte hyperplasia, an early event during the neoplastic process, might begin before liver cirrhosis develops. Genetically obese, leptin-deficient ob/ob mice are models for nonalcoholic fatty liver disease (NAFLD), a type of liver disease that is strongly associated with obesity and type 2 diabetes. Similar to obese, diabetic patients, ob/ob mice have an increased incidence of HCC. However, unlike humans with NAFLD, they rarely, if ever, develop cirrhosis spontaneously. To determine whether the noncirrhotic livers of ob/ob mice with NAFLD exhibit hepatocyte hyperplasia, parameters of proliferation and apoptosis were compared in adult ob/ob mice and their healthy litter mates. Adult ob/ob mice have an increase in liver mass relative to body mass. This hepatomegaly cannot be explained solely by lipid accumulation and is accompanied by significant increases in hepatocyte proliferative activity (as evidenced by increased Erk activation, cell-cycle related gene expression, bromodeoxyuridine incorporation, and hepatic DNA content) with concomitant inhibition of hepatocyte apoptosis (as evidenced by decreased numbers of apoptotic hepatocytes, induction of several antiapoptotic mechanisms, and decreased activation of procaspase 3). Thus, liver hyperplasia is evident at the earliest stage of NAFLD in ob/ob mice, which supports the concept that obesity-related metabolic abnormalities, rather than cirrhosis, initiate the hepatic neoplastic process during obesity.


The protective effect of hypervolemic hemodilution in experimental heatstroke.
The authors tested the hypothesis in a rat model that hypervolemic hemodilution during heatstroke affected the mean arterial pressure (MAP), striatal dopamine (DA) release, and local cerebral blood flow and neuronal damage score in different brain structures. The heatstroke was induced by exposing the urethane-anesthetized rats to an ambient temperature of 42 degrees C. Hypervolemic hemodilution was produced by intravenous administration of 10% human albumin. Relative and absolute blood flow in the corpus striatum were determined using the laser Doppler flowmetry and the autoradiography diffusible tracer technique, respectively. The DA release in the striatum was estimated using the in vivo microdialysis technique. After onset of heatstroke, animals with hypervolemic state alone, produced by saline or heparinized blood injection, displayed higher values of DA release, as well as neuronal damage score in the striatum, hypothalamus, or cortex, but lower values of MAP and blood flow in the striatum, hypothalamus, or cortex compared to normothermic controls. However, the heatstroke-induced arterial hypotension, cerebral ischemia, increased striatal DA overload, and increased neuronal damage score were attenuated by induction of both hypervolemic and hemodilution state with 10% albumin either before or after the onset of heatstroke. In addition, constant infusions of a vasopressor agent phenylephrine (2 microg kg(-1) min(-1)) after the onset of heatstroke failed to maintain appropriate levels of MAP and resulted in no protection against heatstroke. Thus, it appears that the observed benefit of the 10% albumin is secondary to hemodilution and/or maintenance of MAP.


Sox2 plays a role in the induction of amacrine and Müller glial cells in mouse retinal progenitor cells.
The transcription factor Sox2 plays important roles in both human and mouse retinal development. Although loss-of-function mutations in Sox2 have been studied in mice, gain-of-function experiments in the neural retina have been lacking. The detailed expression pattern of Sox2 in the developing mouse retina was examined by immunohistochemistry. Then, Sox2 was expressed in a retinal explant culture prepared from E17 mouse embryos by retrovirus-mediated gene transfer to examine its role in retinal development. In addition, shRNA was used to suppress Sox2 in a retinal explant using a retrovirus-mediated system. Sox2 was expressed throughout the neuroblastic layer in the embryonic retina, but only in the inner nuclear layer in the mature retina. Double immunostaining revealed that Sox2 was expressed in Müller glial cells and in a subset of amacrine cells. Forced expression of Sox2 in a mouse retinal explant culture resulted in the dramatic accumulation of amacrine cells in the inner nuclear layer; in addition, cells expressing amacrine cell markers were also found on the innermost side of the outer nuclear layer. The expression of Pax6, which plays an important role in amacrine cell differentiation, was observed in the Sox2-expressing cells, and Sox2 activated the Pax6 promoter to drive luciferase expression in Y79 cells. A decrease in retinal progenitor cell proliferation accompanied these effects. The suppression of Sox2 expression by shRNA resulted in a decreased number of cells in the inner nuclear layer. Therefore, ectopic Sox2 expression can induce amacrine cells in the mouse retina from stage E17 onward, possibly by facilitating cell cycle exit.


Proteogenomic integration reveals therapeutic targets in breast cancer xenografts.
Recent advances in mass spectrometry (MS) have enabled extensive analysis of cancer proteomes. Here, we employed quantitative proteomics to profile protein expression across 24 breast cancer patient-derived xenograft (PDX) models. Integrated proteogenomic analysis shows positive correlation between expression measurements from transcriptomic and proteomic analyses; further, gene expression-based intrinsic subtypes are largely re-capitulated using non-stromal protein markers. Proteogenomic analysis also validates a number of predicted genomic targets in multiple receptor tyrosine kinases. However, several protein/phosphoprotein events such as overexpression of AKT proteins and ARAF, BRAF, HSP90AB1 phosphosites are not readily explainable by genomic analysis, suggesting that druggable translational and/or post-translational regulatory events may be uniquely diagnosed by MS. Drug treatment experiments targeting HER2 and components of the PI3K pathway supported proteogenomic response predictions in seven xenograft models. Our study demonstrates that MS-based proteomics can identify therapeutic targets and highlights the potential of PDX drug response evaluation to annotate MS-based pathway activities.


[Infrared spectroscopic study of DNA--lipid interactions. DNA compacting on disperse particles].
Within the range of relative humidity (r. h.) 0 to 92% there were obtained IR-spectra (4000--900 cm-1) of undeuterated and deuterated films of rat liver lipids, both in the free state and in the complex with native (nDNA) and heat-denaturated DNA (dDNA). Found peculiarities of IR-spectroscopic realization of the complex-formation are explained qualitatively. One of the peculiarities is hyperchromism of lipid bands and hypochromism of the bands of DNA bases. Lipids are shown to despiralize nDNA in the course of their interaction. A suggestion is advanced concerning the nature of basic contacts which form the complex DNA-lipid. It is found that nDNA unlike dDNA produces an inhibiting effect on lipids oxidation and their subsequent hydrolysis. The mechanism of the phenomenon is discussed. The ideas on DNA compactization resulting from its coiling on different protein and nonprotein particles are developed.


3' RNA processing efficiency plays a primary role in generating termination-competent RNA polymerase II elongation complexes.
In several mammalian transcription units, a transcription termination mechanism in which efficient termination is dependent on the presence of an intact 3' RNA processing site has been identified. The mouse beta maj-globin transcription unit is one such example, in which an intact poly(A) site is required for efficient transcription termination. It is now evident that 3' mRNA processing sites are not always processed with the same efficiency. In this study, we characterized several pre-mRNAs as substrates for the 3' mRNA processing reaction of cleavage and polyadenylation. We then determined whether poly(A) sites which vary in processing efficiency support a poly(A) site-dependent termination event. The level of processing efficiency was determined in vitro by assays measuring the efficiency of the pre-mRNA cleavage event and in vivo by the level of poly(A) site-dependent mRNA and gene product expression generated in transient transfection assays. The beta maj globin pre-mRNA is very efficiently processed. This efficient processing correlates with its function in termination assays using recombinant adenovirus termination vectors in nuclear run-on assays. When the beta maj globin poly(A) site was replaced by the L1 poly(A) site of the adenovirus major late transcription unit (Ad-ml), which is a poor processing substrate, termination efficiency decreased dramatically. When the beta maj globin poly(A) site was replaced by the Ad-ml L3 poly(A) site, which is 10- to 20-fold more efficiently processed than the Ad-ml L1 poly(A) site, termination efficiency remained high. Termination is therefore dependent on the yield of the processing event. We then tested chimeric poly(A) sites containing the L3 core AAUAAA but varied downstream GU-rich elements. The change in downstream GU-rich elements affected processing efficiency in a manner which correlated with termination efficiency. These experiments provide evidence that the efficiency of 3' processing complex formation is directly correlated to the efficiency of RNA polymerase II termination at the 3' end of a mammalian transcription unit.


Reaction screening using a microreactor.
This article discusses the screening of chemical reactions using a microreactor equipped with infrared spectroscopy as online analytics. An esterification reaction has been optimized in continuous mode with the proposed setup. The esterification did not work well due to the material of the microreactor (stainless-steel 316Ti) that catalyzed the decomposition of formic acid. However, despite the occurrence of decomposition, an optimization could be achieved with this system.


Measurement method of nonresonant third-order susceptibility by using off-resonant coherent anti-Stokes Raman spectroscopy.
Nonresonant third-order susceptibilities (χ(nr)) of N(2) and Ar gases by using an off-resonant coherent anti-Stokes Raman spectroscopy of the N(2) Q branch, whose spectral profile is highly sensitive to the value of χ(nr), have been measured to demonstrate the possibility of the present method. The χ(nr) of N(2) and Ar using the off-resonant coherent anti-Stokes Raman spectroscopy were 8.4 ± 0.2 and 9.7 ± 0.8 (× 10(-18)cm(3) /erg/amagat), respectively. These values are in good agreement with those of previous studies. The off-resonant spectra were theoretically calculated by using the Raman linewidth given by a modified exponential gap model, and they were compared with the experimental ones to determine χ(nr). The effects of uncertainties of the Raman linewidth of the N(2) Q branch on the determination of χ(nr) were investigated by changing the parameters in the modified exponential gap model. The determined χ(nr) value changed by only 0.05% with 50% change of the Raman linewidth. The potential advantages of this method are no requirements of high spectral resolution measurements, no accurate Raman linewidth data for each experimental condition, and no optical delay between pump beams.


Harm reduction and 12 steps: complementary, oppositional, or something in-between?
Initially born of the desire to prevent the transmission of HIV among injection drug users, harm reduction presents a relatively new option for assisting individuals who struggle with drug and alcohol use. Twelve-step programs such as Alcoholics Anonymous (AA) are widely recognized as being a representative example of abstinence-based treatment and are often seen as oppositional to harm reduction. The purpose of this study is to examine the ways in which harm reduction workers interpret the relationship between harm reduction and 12-step approaches to treatment. The study draws upon qualitative interviews with 18 staff members from two harm reduction-based substance use treatment programs. (1) Two central themes emerge from the qualitative data: (1) harm reduction and 12-step approaches can be complementary; and (2) 12-step approaches in high-threshold treatment settings may differ significantly from their original philosophy and intent. A third, much less prominent theme reflects some respondents' skepticism about the capacity of the two approaches to work together given the resistance to harm reduction by some in the 12-step community. Complementary conceptualizations of harm reduction and 12-step approaches have the potential to broaden the range of options available to people experiencing substance use problems.


In situ polymerase chain reaction amplification of HIV-1 DNA in brain tissue.
A direct in situ polymerase chain reaction (IS-PCR) assay is described for the detection of HIV-1 proviral DNA in formalin fixed paraffin embedded brain tissue. Biotin-16-dUTP is incorporated during the PCR process and microwave pretreatment of tissue sections ensures that no non-specific incorporation into damaged or nicked genomic DNA occurs. Two methods are compared to detect the biotinylated amplified product, the use of an avidin-biotin-alkaline phosphatase complex (ABC) and the application of tyramide signal amplification (TSA) which allows both chromogenic and fluorescence detection. TSA detection enhances the sensitivity of IS-PCR, permitting fewer PCR cycles and preserving tissue morphology.


An archetypes approach to malaria intervention impact mapping: a new framework and example application.
As both mechanistic and geospatial malaria modeling methods become more integrated into malaria policy decisions, there is increasing demand for strategies that combine these two methods. This paper introduces a novel archetypes-based methodology for generating high-resolution intervention impact maps based on mechanistic model simulations. An example configuration of the framework is described and explored. First, dimensionality reduction and clustering techniques were applied to rasterized geospatial environmental and mosquito covariates to find archetypal malaria transmission patterns. Next, mechanistic models were run on a representative site from each archetype to assess intervention impact. Finally, these mechanistic results were reprojected onto each pixel to generate full maps of intervention impact. The example configuration used ERA5 and Malaria Atlas Project covariates, singular value decomposition, k-means clustering, and the Institute for Disease Modeling's EMOD model to explore a range of three-year malaria interventions primarily focused on vector control and case management. Rainfall, temperature, and mosquito abundance layers were clustered into ten transmission archetypes with distinct properties. Example intervention impact curves and maps highlighted archetype-specific variation in efficacy of vector control interventions. A sensitivity analysis showed that the procedure for selecting representative sites to simulate worked well in all but one archetype. This paper introduces a novel methodology which combines the richness of spatiotemporal mapping with the rigor of mechanistic modeling to create a multi-purpose infrastructure for answering a broad range of important questions in the malaria policy space. It is flexible and adaptable to a range of input covariates, mechanistic models, and mapping strategies and can be adapted to the modelers' setting of choice.


Predictive value of the Luminex single antigen panel for detecting flow cytometry cross-match positivity.
Anti-human leukocyte antigen (HLA) antibodies are a major cause of allograft loss. Solid-phase immunoassays, notably Luminex technology, have lately begun to replace traditional techniques for detecting these antibodies. This platform, however, carries some restrictions in the type of antibodies it detects. For this reason, results using these new technologies must be correlated with results using traditional techniques that have proven clinical significance. We have correlated flow cytometry cross-match (FCXM) outcomes with results from Luminex assays. Serum samples from patients awaiting transplantation who had known anti-HLA antibodies as detected by Luminex were incubated with lymphocytes expressing (a) 1 of the HLA antigens detected by the sera or (b) several of them. Of the 169 T-cell FCXMs we performed, in 92 cases the target cell expressed only 1 of the HLA antigens detected by the serum. The results obtained correlated well with Luminex data (r = 0.84). A cutoff mean fluorescence intensity value of 6,500 for the Luminex single antigen assay yielded a sensitivity of 85% and specificity of 82% for detecting a positive FCXM. In the other 77 cases, the target cell expressed 2 or more of the HLA antigens detected by the serum. In this situation, the same cutoff proved a useful tool for differentiating negative from positive FCXMs.


Biochemical and crystallographic analyses of a portal mutant of the adipocyte lipid-binding protein.
A number of crystallographic studies of the adipocyte lipid-binding protein have established that the fatty acid-binding site is within an internalized water-filled cavity. The same studies have also suggested the existence of a region physically distinct from the fatty acid-binding site which connects the cavity of the protein with the external solvent, hereafter referred to as the portal. In an effort to examine the portal region, we have used site-directed mutagenesis to introduce the mutations V32D/F57H into the murine ALBP cDNA. Mutant protein has been isolated, crystallized, and its stability and binding properties studied by biochemical methods. As assessed by guanidine-HCl denaturation, the mutant form exhibited a slight overall destabilization relative to the wild-type protein under both acid and alkaline conditions. Accessibility to the cavity in both the mutant and wild-type proteins was observed by stopped-flow analysis of the modification of a cavity residue, Cys117, by the sulfhydryl reactive agent 5, 5'-dithiobis(2-nitrobenzoic acid) at pH 8.5. Cys117 of V32D/F57H ALBP was modified 7-fold faster than the wild-type protein. The ligand binding properties of both the V32D/F57H mutant and wild-type proteins were analyzed using a fluorescent probe at pH 6.0 and 8.0. The apparent dissociation constants for 1-anilinonaphthalene-8-sulfonic acid were approximately 9-10-fold greater than the wild-type protein, independent of pH. In addition, there is a 6-fold increase in the Kd for oleic acid for the portal mutant relative to the wild-type at pH 8.0. To study the effect of pH on the double mutant, it was crystallized and analyzed in two distinct space groups at pH 4.5 and 6.4. While in general the differences in the overall main chain conformations are negligible, changes were observed in the crystallographic structures near the site of the mutations. At both pH values, the mutant side chains are positioned somewhat differently than in wild-type protein. To ensure that the mutations had not altered ionic conditions near the binding site, the crystallographic coordinates were used to monitor the electrostatic potentials from the head group site to the positions near the portal region. The differences in the electrostatic potentials were small in all regions, and did not explain the differences in ligand affinity. We present these results within the context of fatty acid binding and suggest lipid association is more complex than that described within a single equilibrium event.


Inactivation dynamics of Salmonella enterica, Listeria monocytogenes, and Escherichia coli O157:H7 in wash water during simulated chlorine depletion and replenishment processes.
Maintaining effective sanitizer concentration is of critical importance for preventing pathogen survival and transference during fresh-cut produce wash operation and for ensuring the safety of finished products. However, maintaining an adequate level of sanitizer in wash water can be challenging for processors due to the large organic load in the wash system. In this study, we investigated how the survival of human pathogens was affected by the dynamic changes in water quality during chlorine depletion and replenishment in simulated produce washing operations. Lettuce extract was added incrementally into water containing pre-set levels of free chlorine to simulate the chlorine depletion process, and sodium hypochlorite was added incrementally into water containing pre-set levels of lettuce extract to simulate chlorine replenishment. Key water quality parameters were closely monitored and the bactericidal activity of the wash water was evaluated using three-strain cocktails of Escherichia coli O157:H7, Salmonella enterica, and Listeria monocytogenes. In both chlorine depletion and replenishment processes, no pathogen survival was observed when wash water free chlorine level was maintained above 3.66 mg/L, irrespective of the initial free chlorine levels (10, 50, 100 and 200 mg/L) or organic loading (chemical oxidation demand levels of 0, 532, 1013 and 1705 mg/L). At this free chlorine concentration, the measured ORP was 843 mV and pH was 5.12 for the chlorine depletion process; the measured ORP was 714 mV and pH was 6.97 for the chlorine replenishment process. This study provides quantitative data needed by the fresh-cut produce industry and the regulatory agencies to establish critical operational control parameters to prevent pathogen survival and cross-contamination during fresh produce washing.


FBXO7 Y52C polymorphism as a potential protective factor in Parkinson's disease.
Mutations in the F-box only protein 7 gene (FBXO7), the substrate-specifying subunit of SCF E3 ubiquitin ligase complex, cause Parkinson's disease (PD)-15 (PARK15). To identify new variants, we sequenced FBXO7 cDNA in 80 Taiwanese early onset PD patients (age at onset ≤ 50) and only two known variants, Y52C (c.155A>G) and M115I (c.345G>A), were found. To assess the association of Y52C and M115I with the risk of PD, we conducted a case-control study in a cohort of PD and ethnically matched controls. There was a nominal difference in the Y52C G allele frequency between PD and controls (p = 0.045). After combining data from China [1], significant difference in the Y52C G allele frequency between PD and controls (p = 0.012) and significant association of G allele with decreased PD risk (p = 0.017) can be demonstrated. Upon expressing EGFP-tagged Cys52 FBXO7 in cells, a significantly reduced rate of FBXO7 protein decay was observed when compared with cells expressing Tyr52 FBXO7. In silico modeling of Cys52 exhibited a more stable feature than Tyr52. In cells expressing Cys52 FBXO7, the level of TNF receptor-associated factor 2 (TRAF2) was significantly reduced. Moreover, Cys52 FBXO7 showed stronger interaction with TRAF2 and promoted TRAF2 ubiquitination, which may be responsible for the reduced TRAF2 expression in Cys52 cells. After induced differentiation, SH-SY5Y cells expressing Cys52 FBXO7 displayed increased neuronal outgrowth. We therefore hypothesize that Cys52 variant of FBXO7 may contribute to reduced PD susceptibility in Chinese.


Chronotype variation drives night-time sentinel-like behaviour in hunter-gatherers.
Sleep is essential for survival, yet it also represents a time of extreme vulnerability to predation, hostile conspecifics and environmental dangers. To reduce the risks of sleeping, the sentinel hypothesis proposes that group-living animals share the task of vigilance during sleep, with some individuals sleeping while others are awake. To investigate sentinel-like behaviour in sleeping humans, we investigated activity patterns at night among Hadza hunter-gatherers of Tanzania. Using actigraphy, we discovered that all subjects were simultaneously scored as asleep for only 18 min in total over 20 days of observation, with a median of eight individuals awake throughout the night-time period; thus, one or more individuals was awake (or in light stages of sleep) during 99.8% of sampled epochs between when the first person went to sleep and the last person awoke. We show that this asynchrony in activity levels is produced by chronotype variation, and that chronotype covaries with age. Thus, asynchronous periods of wakefulness provide an opportunity for vigilance when sleeping in groups. We propose that throughout human evolution, sleeping groups composed of mixed age classes provided a form of vigilance. Chronotype variation and human sleep architecture (including nocturnal awakenings) in modern populations may therefore represent a legacy of natural selection acting in the past to reduce the dangers of sleep.


Pelvic reconstruction for severe periacetabular metastatic disease.
The functional outcome of acetabular reconstruction in 21 patients with severe acetabular deficiency was examined. The surgical indications were pain, immobility, and loss of indepedence. Surgical reconstruction included curettage, placement of an acetabular roof ring with screws crossing the defect, cementation of the defect, and cemented hip arthroplasty. Although three patients died within the first month, the mean postoperative survival was 14.5 +/- 4.0 months, with a 10.4 +/- 3.0-month period of independent living. Statistically significant postoperative improvements were seen in pain and mobility scores. Using the Musculoskeletal Tumor Society score for the lower extremity, a mean postoperative score of 14.2 of a total 30 was calculated. Low scores were attributed to the patients' overall disease, including the requirement of continued narcotic pain control, occupational restriction, and the presence of Trendelenburg limp. Surgical complications occurred in six patients and included three early postoperative deaths, two related to poor preoperative pulmonary function; two dislocations, one related to a late infection; one femoral nerve palsy; and one foot drop. Although associated with a high complication rate, acetabular reconstruction for metastatic disease can lead to marked improvement in pain control, function, independence, and quality of life in carefully selected patients.


Relationship between radiosensitivity and Nrf2 target gene expression in human hematopoietic stem cells.
NFE2-related factor 2 (Nrf2), which belongs to the cap "n" collar family of basic region leucine zipper transcription factors, is a key protein in the coordinated transcriptional induction of expression of various antioxidant genes. The purpose of this study was to analyze the expression of Nrf2 target genes, such as heme oxygenase 1 (HO-1), ferritin heavy polypeptide 1 (FTH1), NAD(P)H dehydrogenase, quinone 1 (NQO1), glutamate-cysteine ligase catalytic subunit, glutamate-cysteine ligase modifier subunit, glutathione reductase (GSR) and thioredoxin reductase 1 (TXNRD1), after X irradiation of CD34(+) cells that were prepared from human placental/umbilical cord blood hematopoietic stem cells (HSCs). We evaluated the relationship between radiosensitivity and expression of Nrf2 target genes in HSCs. The number of colony-forming cells derived from 2-Gy-irradiated HSCs decreased to approximately 20% of the nonirradiated control. At the same time, the mRNA expression of HO-1, FTH1, NQO1, GSR and TXNRD1 was significantly increased after X irradiation. A statistically significant negative correlation was observed between the surviving fraction of HSCs and the intrinsic NQO1 mRNA expression, indicating that HSCs in which NQO1 mRNA levels are low may also be radioresistant. The present results suggest that the antioxidant system associated with Nrf2 is involved in the radiosensitivity of HSCs.


The screw-helical voltage gating of ion channels.
In the voltage-gated ion channels of every animal, whether they are selective for K+, Na+ or Ca2+, the voltage sensors are the S4 transmembrane segments carrying four to eight positive charges always separated by two uncharged residues. It is proposed that they move across the membrane in a screw-helical fashion in a series of three or more steps that each transfer a single electronic charge. The unit steps are stabilized by ion pairing between the mobile positive charges and fixed negative charges, of which there are invariably two located near the inner ends of segments S2 and S3 and a third near the outer end of either S2 or S3. Opening of the channel involves three such steps in each domain.


Characterization of Novel Antimalarial Compound ACT-451840: Preclinical Assessment of Activity and Dose-Efficacy Modeling.
Artemisinin resistance observed in Southeast Asia threatens the continued use of artemisinin-based combination therapy in endemic countries. Additionally, the diversity of chemical mode of action in the global portfolio of marketed antimalarials is extremely limited. Addressing the urgent need for the development of new antimalarials, a chemical class of potent antimalarial compounds with a novel mode of action was recently identified. Herein, the preclinical characterization of one of these compounds, ACT-451840, conducted in partnership with academic and industrial groups is presented. The properties of ACT-451840 are described, including its spectrum of activities against multiple life cycle stages of the human malaria parasite Plasmodium falciparum (asexual and sexual) and Plasmodium vivax (asexual) as well as oral in vivo efficacies in two murine malaria models that permit infection with the human and the rodent parasites P. falciparum and Plasmodium berghei, respectively. In vitro, ACT-451840 showed a 50% inhibition concentration of 0.4 nM (standard deviation [SD]: ± 0.0 nM) against the drug-sensitive P. falciparum NF54 strain. The 90% effective doses in the in vivo efficacy models were 3.7 mg/kg against P. falciparum (95% confidence interval: 3.3-4.9 mg/kg) and 13 mg/kg against P. berghei (95% confidence interval: 11-16 mg/kg). ACT-451840 potently prevented male gamete formation from the gametocyte stage with a 50% inhibition concentration of 5.89 nM (SD: ± 1.80 nM) and dose-dependently blocked oocyst development in the mosquito with a 50% inhibitory concentration of 30 nM (range: 23-39). The compound's preclinical safety profile is presented and is in line with the published results of the first-in-man study in healthy male participants, in whom ACT-451840 was well tolerated. Pharmacokinetic/pharmacodynamic (PK/PD) modeling was applied using efficacy in the murine models (defined either as antimalarial activity or as survival) in relation to area under the concentration versus time curve (AUC), maximum observed plasma concentration (Cmax), and time above a threshold concentration. The determination of the dose-efficacy relationship of ACT-451840 under curative conditions in rodent malaria models allowed prediction of the human efficacious exposure. The dual activity of ACT-451840 against asexual and sexual stages of P. falciparum and the activity on P. vivax have the potential to meet the specific profile of a target compound that could replace the fast-acting artemisinin component and harbor additional gametocytocidal activity and, thereby, transmission-blocking properties. The fast parasite reduction ratio (PRR) and gametocytocidal effect of ACT-451840 were recently also confirmed in a clinical proof-of-concept (POC) study.


Analysis of electro-optic crystal-based Fabry-Perot etalons for high-speed spatial light modulators.
The use of electro-optic (EO) crystal-based Fabry-Perot modulators (FPMs) as high-speed spatial light modulators is proposed. The FPMs operate with an extremely low drive voltage and a high extinction ratio. It is revealed by analysis of both the linear EO effect and the inverse piezoelectric effect of various EO crystals that three kinds of crystal configuration are suitable as FPMs. One of these is applicable to isotropic crystals, point groups 23 and (-)43m, and the others are better suited for uniaxial EO crystals, point groups (-)42m and 3m. Typical EO crystals suitable as FPMs are ferroelectric crystals such as LiNbO3, LiTaO3, and LiIO3 and sillenite compounds such as Bi12SiO20 and Bi12GeO20 as well as compound semiconductors such as GaAs and GaP.


Individual differences in children's event memory reports and the narrative elaboration technique.
Children between 7 and 8 years old took part in a staged event at school and 1 week later were assessed using a short form of the Wechsler Intelligence scale for children (third edition) and measures of metamemory, narrative ability, and socioeconomic status. Two weeks following the event, children either received narrative elaboration training (NET; K.J. Saywitz & L. Snyder, 1996) and were prompted with the four NET cue cards at interview; received verbal prompts corresponding to the cue card categories, but without prior training; or were presented with the cards at interview without prior training. Children given verbal labels as prompts recalled as much information as children who received NET training and cue cards. Measures of intelligence were predictive of amount recalled for cards-only children but not for the other 2 groups, indicating that differences in recall between low- and high-IQ groups were attenuated when recall was supported by NET training or verbal prompting.


Oncologists' protocol and coping strategies in dealing with patient loss.
To identify what protocol and coping strategies oncologists turn to cope with patient loss, the authors interviewed 20 oncologists at 3 hospitals. Using the grounded theory method, findings revealed that their protocol may include meeting with families, participating in bereavement rituals, making a phone call, or sending a condolence card. Coping strategies included social support, activity-oriented coping, turning to faith, compartmentalization, and withdrawing from patients and families. The authors conclude by offering implications from this research on how to address oncologists' grief over patient loss in institutional settings in order to improve bereavement care for families and enhance oncologists' quality of life.


The Functional Utilization of Propulsive Capacity During Human Walking.
Aging and many gait pathologies are characterized by reduced propulsive forces and ankle moment and power generation during trailing leg push-off in walking. Despite those changes, we posit that many individuals retain an underutilized reserve for enhancing push-off intensity during walking that may be missed using conventional dynamometry. By using a maximum ramped impeding force protocol and maximum speed walking, we gained mechanistic insight into the factors that govern push-off intensity and the available capacity thereof during walking in young subjects. We discovered in part that young subjects walking at their preferred speed retain a reserve capacity for exerting larger propulsive forces of 49%, peak ankle power of 43%, and peak ankle moment of 22% during push-off-the latter overlooked by maximum isometric dynamometry. We also provide evidence that these reserve capacities are governed at least in part by the neuromechanical behavior of the plantarflexor muscles, at least with regard to ankle moment generation. We envision that a similar paradigm used to quantify propulsive reserves in older adults or people with gait pathology would empower the more discriminate and personalized prescription of gait interventions seeking to improve push-off intensity and thus walking performance.


Children's cognitive recovery after day-case general anesthesia: a randomized trial of propofol or isoflurane for dental procedures.
Postoperative cognitive dysfunction (POCD) is well established in adults but has been underinvestigated in children. As day-case procedures are increasingly common, it is important to establish whether children suffer significant POCD. Pediatric POCD has been associated with several intravenous and inhalation anesthetics, but isoflurane has not been studied. As evidence indicates superior recovery after propofol, the study compared POCD after propofol or isoflurane anesthesia. To compare the effects of propofol versus isoflurane upon children's POCD. Fifty-eight children aged 5-14 years were randomized to propofol (total intravenous anesthesia) or isoflurane for day-case dental procedures. Reaction time (RT), verbal and visual memory, psychomotor coordination, and attention were assessed preoperatively, prior to discharge and at 24 h. Reaction time and psychomotor control were impaired postoperatively in both groups but recovered at 24 h. Delayed verbal recall was significantly impaired only after propofol. Both groups had significant impairment of visual memory postoperatively and at 24 h, and of recognition memory postoperatively only. Propofol and isoflurane exert similar adverse effects on RT, psychomotor coordination, and visual memory. Selective impairment of verbal recall by propofol is consistent with adult evidence of the drug's effect on retrieval. The enduring postoperative impairment of memory has implications for instructions to parents and caregivers for the safety and well-being of children in the 24 h after day-case anesthesia with propofol and isoflurane.


Ascorbic acid regulates osterix expression in osteoblasts by activation of prolyl hydroxylase and ubiquitination-mediated proteosomal degradation pathway.
Mouse genetic studies reveal that ascorbic acid (AA) is essential for osteoblast (OB) differentiation and that osterix (Osx) was a key downstream target of AA action in OBs. To determine the molecular pathways for AA regulation of Osx expression, we evaluated if AA regulates Osx expression by regulating production and/or actions of local growth factors and extracellular matrix (ECM) proteins. Inhibition of actions of IGFs by inhibitory IGFBP-4, BMPs by noggin, and ECM-mediated integrin signaling by RGD did not block AA effects on Osx expression in OBs. Furthermore, blockade of components of MAPK signaling pathway had no effect on AA-induced Osx expression. Because AA is required for prolyl hydroxylase domain (PHD) activity and because PHD-induced prolyl-hydroxylation targets proteins to proteosomal degradation, we next tested if AA effect on Osx expression involves activation of PHD to hydroxylate and induce ubiquitin-proteosome-mediated degradation of transcriptional repressor(s) of Osx gene. Treatment of OBs with dimethyloxallyl glycine and ethyl 3, 4-dihydroxybenzoate, known inhibitors of PHD, completely blocked AA effect on Osx expression and OB differentiation. Knockdown of PHD2 expression by Lentivirus-mediated shRNA abolished AA-induced Osx induction and alkaline phosphatase activity. Furthermore, treatment of OBs with MG115, inhibitor of proteosomal degradation, completely blocked AA effects on Osx expression. Based on these data, we conclude that AA effect on Osx expression is mediated via a novel mechanism that involves PHD2 and proteosomal degradation of a yet to be identified transcriptional repressor that is independent of BMP, IGF-I, or integrin-mediated signaling in mouse OBs.


SERIES: eHealth in primary care. Part 2: Exploring the ethical implications of its application in primary care practice.
Background: eHealth promises to increase self-management and personalised medicine and improve cost-effectiveness in primary care. Paired with these promises are ethical implications, as eHealth will affect patients' and primary care professionals' (PCPs) experiences, values, norms, and relationships.Objectives: We argue what ethical implications related to the impact of eHealth on four vital aspects of primary care could (and should) be anticipated.Discussion: (1) EHealth influences dealing with predictive and diagnostic uncertainty. Machine-learning based clinical decision support systems offer (seemingly) objective, quantified, and personalised outcomes. However, they also introduce new loci of uncertainty and subjectivity. The decision-making process becomes opaque, and algorithms can be invalid, biased, or even discriminatory. This has implications for professional responsibilities and judgments, justice, autonomy, and trust. (2) EHealth affects the roles and responsibilities of patients because it can stimulate self-management and autonomy. However, autonomy can also be compromised, e.g. in cases of persuasive technologies and eHealth can increase existing health disparities. (3) The delegation of tasks to a network of technologies and stakeholders requires attention for responsibility gaps and new responsibilities. (4) The triangulate relationship: patient-eHealth-PCP requires a reconsideration of the role of human interaction and 'humanness' in primary care as well as of shaping Shared Decision Making.Conclusion: Our analysis is an essential first step towards setting up a dedicated ethics research agenda that should be examined in parallel to the development and implementation of eHealth. The ultimate goal is to inspire the development of practice-specific ethical recommendations.


Microbes are not bound by sociobiology: response to Kümmerli and Ross-Gillespie (2013).
In recent years, sociobiology has been extended to microorganisms. Viewed through this lens, the microbial world is replete with cooperative behaviors. However, little attention has been paid to alternate hypotheses, making many studies self-confirming. Somewhat apart is a recent analysis of pyoverdin production-a paradigmatic public good and social trait-by Pseudomonas, which has revealed discord between predictions arising from sociobiology and the biology of microbes. This led the authors, Zhang and Rainey (Z&R), to question the generality of the conclusion that pyoverdin is a social trait, and to question the fit between the sociobiology framework and microbiology. This has unsettled Kümmerli and Ross-Gillespie (K&R), who in a recent "Technical Comment" assert that arguments presented by Z&R are flawed, their experiments technically mistaken, and their understanding of social evolution theory naive. We demonstrate these claims to be without substance and show the conclusions of K&R to be based on a lack of understanding of redox chemistry and on misinterpretation of data. We also point to evidence of cherry-picking and raise the possibility of confirmation bias. Finally, we emphasize that the sociobiology framework applied to microbes is a hypothesis that requires rigorous and careful appraisal.


Evidence of synergy between Thy-1 and CD3/TCR complex in signal delivery to murine thymocytes for cell death.
The potential role of Thy-1 in CD3/TCR complex-mediated signal delivery to murine thymocytes was studied. Ag-mimicking cross-linked anti-CD3 mAb stimulated suspension of thymocytes from adult (6 to 8 wk old) mice for a brisk free cytoplasmic calcium ion ([Ca2+]i) rise, low level of inositol phosphate production, and marginal increase in tyrosine-specific phosphorylation of 110/120-kDa and 40-kDa cellular proteins. Weak but sustained [Ca2+]i rise, low inositol phosphate production, and weak protein tyrosine phosphorylation were also induced by the cross-linked anti-Thy-1 mAb that mimicked the putative natural ligand. The signal delivered via either of these two pathways was however insufficient for definitively promoting cell death and DNA fragmentation in the adult thymocytes. Here we demonstrated that anti-Thy-1 mAb synergized with anti-CD3 mAb for inducing a long-lasting prominent [Ca2+]i rise, definite inositol 1,4,5-triphosphate and inositol 1,3,4,5-tetrakiphosphate production, and extensive tyrosine-specific phosphorylation of 110/120-, 92-, 75-, and 40-kDa proteins, which resulted in marked promotion of cell death and DNA fragmentation in the adult thymocytes. This unique anti-Thy-1 antibody activity was confirmed to be directed to glycosylphosphatidylinositol-anchored Thy-1, and was distinguished from the known anti-L3T4 activity that augmented the CD3-mediated signal transduction in a different manner. The synergistic actions of anti-CD3 and anti-Thy-1 mAb obligatorily required the cross-linking of the two mAb together. The anti-CD3 and anti-Thy-1 mAb cross-linked together acted on immature thymocytes from newborn (less than 24 h after birth) mice for rather more extensive promotion of protein tyrosine phosphorylation and cell death. In addition, they affected peripheral T lymphocytes for accelerating protein tyrosine phosphorylation but not cell death. These results suggest a novel function of glycosylphosphatidylinositol-anchored Thy-1 as a possible unique intrathymic intensifier of the CD3/TCR complex-delivered signal for negative thymocyte selection.


Induction of Exaggerated Cytokine Production in Human Peripheral Blood Mononuclear Cells by a Recombinant SARS-CoV-2 Spike Glycoprotein S1 and Its Inhibition by Dexamethasone.
An understanding of the pathological inflammatory mechanisms involved in SARS-CoV-2 virus infection is necessary in order to discover new molecular pharmacological targets for SARS-CoV-2 cytokine storm. In this study, the effects of a recombinant SARS-CoV-2 spike glycoprotein S1 was investigated in human peripheral blood mononuclear cells (PBMCs). Stimulation of PBMCs with spike glycoprotein S1 (100 ng/mL) resulted in significant elevation in the production of TNFα, IL-6, IL-1β and IL-8. However, pre-treatment with dexamethasone (100 nM) caused significant reduction in the release of these cytokines. Further experiments revealed that S1 stimulation of PBMCs increased phosphorylation of NF-κB p65 and IκBα, and IκBα degradation. DNA binding of NF-κB p65 was also significantly increased following stimulation with spike glycoprotein S1. Treatment of PBMCs with dexamethasone (100 nM) or BAY11-7082 (1 μM) resulted in inhibition of spike glycoprotein S1-induced NF-κB activation. Activation of p38 MAPK by S1 was blocked in the presence of dexamethasone and SKF 86002. CRID3, but not dexamethasone pre-treatment, produced significant inhibition of S1-induced activation of NLRP3/caspase-1. Further experiments revealed that S1-induced increase in the production of TNFα, IL-6, IL-1β and IL-8 was reduced in the presence of BAY11-7082 and SKF 86002, while CRID3 pre-treatment resulted in the reduction of IL-1β production. These results suggest that SARS-CoV-2 spike glycoprotein S1 stimulated PBMCs to release pro-inflammatory cytokines through mechanisms involving activation of NF-κB, p38 MAPK and NLRP3 inflammasome. It is proposed that the clinical benefits of dexamethasone in COVID-19 are possibly due to its anti-inflammatory activity in reducing SARS-CoV-2 cytokine storm.