Progressive retinal atrophy in the Abyssinian cat. Electron microscopy.
Seven adult Abyssinian cats at different stages of a recessively inherited retinal degenerative disease (progressive retinal atrophy) were studied ultrastructurally. At the stage of early disease, in 2-yr-old cats, disorganized and vesiculated discs were found in less than half of the rod outer segments in the periphery, while similar changes were seen in the central retina only infrequently or in patches. Cones appeared normal in all areas of the retina at the early stage. With progression of disease, the lesions were more advanced in all areas of the retina, and involved both rods and cones, with the most severe alterations found in the midperiphery. At the advanced stage, in a 6-yr-old cat, both rods and cones were lost, the inner nuclear layer thus being separated from the pigment epithelium by Müller cell processes and a few remaining outer plexiform processes only. Remnants of photoreceptor outer and inner segments, macrophages, and what appeared to be displaced photoreceptor cell nuclei could be found occasionally in the subretinal space, however. Clumps of pigment granules were often observed in the photoreceptor layer in the non-tapetal fundus. The pigment epithelium remained morphologically intact as a single layer of uninterrupted cells throughout the disease process, as did tapetal cells and choriocapillaris. There was no difference in the severity of disease between the peripheral tapetal and non-tapetal fundus. In the inner retina, only minor alterations were observed. These changes appeared at a later time than photoreceptor degeneration, and were considered secondary to the latter.


Trends in incidence of anal cancer in Denmark.
To study long term trends in incidence of anal cancer in a well monitored, unselected population. Descriptive epidemiological study based on data from the Danish Cancer Registry. Denmark, 1943-87. Time related changes in anal cancer incidence according to sex, age, birth cohort, urban or rural residence, and marital status. The incidence of anal cancer remained fairly constant in the period 1943-57 and was similar for men and women, but it increased 1.5-fold among men and nearly tripled among women thereafter. Among men the incidence increased from 0.25 per 100,000 population (world standardised) in 1958-62 to 0.38 in 1983-7 (p = 0.01) and among women from 0.28 to 0.74 (p < 0.01). The greatest increase was among residents of the capital (Copenhagen). During 1943-87 age specific trends increased in young and middle aged men and in all age groups among women. Men with anal cancer were significantly more likely throughout the study period to be unmarried than were patients with cancer of the colon (adjusted odds ratio 2.7; 95% confidence interval 2.0 to 3.6) and stomach (2.1; 1.5 to 2.8), but no association with marital status was found among women. The distribution and incidence of anal cancer have changed appreciably since around 1960, especially among women, which indicates important aetiological changes. Changes in sexual behaviour may have facilitated the spread of a transmittable agent of aetiological importance. It has recently been suggested that cigarette smoking promotes anal cancer, and this finds indirect support in the synchronism between changes in anal cancer incidence and heavy smoking behaviour. Factors associated with homosexuality are likely to explain some of the cases among men.


Using Covert Response Activation to Test Latent Assumptions of Formal Decision-Making Models in Humans.
Most decisions that we make build upon multiple streams of sensory evidence and control mechanisms are needed to filter out irrelevant information. Sequential sampling models of perceptual decision making have recently been enriched by attentional mechanisms that weight sensory evidence in a dynamic and goal-directed way. However, the framework retains the longstanding hypothesis that motor activity is engaged only once a decision threshold is reached. To probe latent assumptions of these models, neurophysiological indices are needed. Therefore, we collected behavioral and EMG data in the flanker task, a standard paradigm to investigate decisions about relevance. Although the models captured response time distributions and accuracy data, EMG analyses of response agonist muscles challenged the assumption of independence between decision and motor processes. Those analyses revealed covert incorrect EMG activity ("partial error") in a fraction of trials in which the correct response was finally given, providing intermediate states of evidence accumulation and response activation at the single-trial level. We extended the models by allowing motor activity to occur before a commitment to a choice and demonstrated that the proposed framework captured the rate, latency, and EMG surface of partial errors, along with the speed of the correction process. In return, EMG data provided strong constraints to discriminate between competing models that made similar behavioral predictions. Our study opens new theoretical and methodological avenues for understanding the links among decision making, cognitive control, and motor execution in humans. Sequential sampling models of perceptual decision making assume that sensory information is accumulated until a criterion quantity of evidence is obtained, from where the decision terminates in a choice and motor activity is engaged. The very existence of covert incorrect EMG activity ("partial error") during the evidence accumulation process challenges this longstanding assumption. In the present work, we use partial errors to better constrain sequential sampling models at the single-trial level.


Dynamic optical arbitrary waveform generation and measurement.
We introduce a dynamic optical arbitrary waveform generation (OAWG) technique that produces bandwidth scalable, continuous waveforms of near perfect fidelity. Additionally, OAWG's complement, real-time arbitrary optical waveform measurement (OAWM) is discussed. These approaches utilize gigahertz-bandwidth electronics to generate, or measure, truly arbitrary and dynamic optical waveforms scalable to terahertz bandwidths and infinite record lengths. We describe the theory, algorithms and enabling technologies necessary to calculate and produce a set of spectral modulations that create continuous, high-fidelity waveforms in the presence of spectral filtering from multiplexers.


RNA silencing proteins and small RNAs in oomycete plant pathogens and biocontrol agents.
Oomycetes cause several damaging diseases of plants and animals, and some species also act as biocontrol agents on insects, fungi, and other oomycetes. RNA silencing is increasingly being shown to play a role in the pathogenicity of Phytophthora species, either through trans-boundary movement of small RNAs (sRNAs) or through expression regulation of infection promoting effectors. To gain a wider understanding of RNA silencing in oomycete species with more diverse hosts, we mined genome assemblies for Dicer-like (DCL), Argonaute (AGO), and RNA dependent RNA polymerase (RDRP) proteins from Phytophthora plurivora, Ph. cactorum, Ph. colocasiae, Pythium oligandrum, Py. periplocum, and Lagenidium giganteum. Moreover, we sequenced small RNAs from the mycelium stage in each of these species. Each of the species possessed a single DCL protein, but they differed in the number and sequence of AGOs and RDRPs. SRNAs of 21nt, 25nt, and 26nt were prevalent in all oomycetes analyzed, but the relative abundance and 5' base preference of these classes differed markedly between genera. Most sRNAs mapped to transposons and other repeats, signifying that the major role for RNA silencing in oomycetes is to limit the expansion of these elements. We also found that sRNAs may act to regulate the expression of duplicated genes. Other sRNAs mapped to several gene families, and this number was higher in Pythium spp., suggesting a role of RNA silencing in regulating gene expression. Genes for most effector classes were the source of sRNAs of variable size, but some gene families showed a preference for specific classes of sRNAs, such as 25/26 nt sRNAs targeting RxLR effector genes in Phytophthora species. Novel miRNA-like RNAs (milRNAs) were discovered in all species, and two were predicted to target transcripts for RxLR effectors in Ph. plurivora and Ph. cactorum, indicating a putative role in regulating infection. Moreover, milRNAs from the biocontrol Pythium species had matches in the predicted transcriptome of Phytophthora infestans and Botrytis cinerea, and L. giganteum milRNAs matched candidate genes in the mosquito Aedes aegypti. This suggests that trans-boundary RNA silencing may have a role in the biocontrol action of these oomycetes.


Interlaboratory study evaluating quantitation of antibodies to Haemophilus influenzae type b polysaccharide by enzyme-linked immunosorbent assay.
An interlaboratory study was conducted to determine whether an enzyme-linked immunosorbent assay (ELISA) with an antigen preparation composed of various-sized fragments of Haemophilus influenzae type b polysaccharide conjugated to human serum albumin could be standardized across laboratories and whether the ELISA-derived results from different laboratories are equivalent to those obtained by the standard radioactive antigen binding assay (RABA) for quantitation of anti-H, influenzae type b polysaccharide antibodies. Twenty coded human serum samples were quantitated by ELISA in 11 laboratories and by RABA in 5 laboratories. The mean RABA-derived values served as the basis for all comparisons. While the overall correspondence of antibody values between the two methods was good, significant differences were found among some of the 11 ELISA data sets and among the mean RABA values. Seven laboratories generated higher ELISA antibody values for low-titered sera. Four laboratories generated antibody concentrations that were not statistically different between the two assay methods. The results therefore indicate that the ELISA can tolerate substantial variations in protocol, such as the use of different plates and different antibody reagents, without affecting the quantitation of serum antibodies. However, attention should be focused on low-titered sera, as some assay conditions may yield spurious results. This ELISA is a serologic assay which can serve as an alternative to the RABA for quantitation of antibodies to H. influenzae type h polysaccharide.


One or two membranes? Diderm Firmicutes challenge the Gram-positive/Gram-negative divide.
How, when and why the transition between cell envelopes with one membrane (Gram-positives or monoderms) and two (Gram-negative or diderms) occurred in Bacteria is a key unanswered question in evolutionary biology. Different hypotheses have been put forward, suggesting that either the monoderm or the diderm phenotype is ancestral. The existence of diderm members in the classically monoderm Firmicutes challenges the Gram-positive/Gram-negative divide and provides a great opportunity to tackle the issue. In this review, we present current knowledge on the diversity of bacterial cell envelopes, including these atypical Firmicutes. We discuss how phylogenomic analysis supports the hypothesis that the diderm cell envelope architecture is an ancestral character in the Firmicutes, and that the monoderm phenotype in this phylum arose multiple times independently by loss of the outer membrane. Given the overwhelming distribution of diderm phenotypes with respect to monoderm ones, this scenario likely extends to the ancestor of all bacteria. Finally, we discuss the recent development of genetic tools for Veillonella parvula, a diderm Firmicute member of the human microbiome, which indicates it as an emerging new experimental model to investigate fundamental aspects of the diderm/monoderm transition.


Continuous infusion of tracer norepinephrine may miscalculate unidirectional nerve uptake of norepinephrine in humans.
In order to evaluate uptake kinetics of norepinephrine (NE) in different tissues, a catheterization study was performed in control subjects (n = 6) and patients with enhanced sympathetic nervous activity (cirrhosis, n = 12) during constant intravenous infusion of L[3H]norepinephrine ([3H]NE) for 75 minutes. In spite of a higher NE spillover from kidneys in patients compared with controls (82 vs. 49 ng/min, p less than 0.01), renal extraction ratios of [3H]NE were similar in the two groups (0.33 vs. 0.32, NS), and no significant change was observed during the time of infusion. In contrast, liver-intestine extraction ratios of [3H]NE decreased significantly and equally with infusion time in patients (from 0.57 to 0.44, p less than 0.01) and controls (from 0.59 to 0.46, p less than 0.01). This was observed despite the fact that spillover of NE from this vascular bed was observed only in patients with cirrhosis and not in controls (41 vs. -5 ng/min, p less than 0.02). In the lower limb, net release of NE was similar in patients and controls, and extraction ratios of [3H]NE decreased almost equally with infusion time (from 0.35 to 0.30, p less than 0.01 and from 0.40 to 0.24, p less than 0.1, respectively). Whole-body clearance of [3H]NE decreased over time in patients (-6%, p less than 0.01) and controls (-20%, p less than 0.01), but significant difference was not observed between the groups. We conclude that failure to attain a steady state with respect to [3H]NE removal was demonstrated in areas of large tissue volume relative to blood flow.(ABSTRACT TRUNCATED AT 250 WORDS)


Cancer patients' experiences with nature: Normalizing dichotomous realities.
To explore cancer patients' subjective experiences with nature in order to examine the relevance of nature-based care opportunities in cancer care contexts. The rationale was to describe the underlying mechanisms of this interaction and produce translatable knowledge. Qualitative research design informed by grounded theory. Sampling was initially convenience and then theoretical. Competent adults with any cancer diagnosis were eligible to participate in a semi-structured interview exploring views about the role of nature in their lives. Audio-recorded and transcribed interviews were analyzed using inductive, cyclic, and constant comparative analysis. Twenty cancer patients (9 female) reported detailed description about their experiences with nature from which a typology of five common nature interactions emerged. A theory model was generated constituting a core category and two inter-related themes explaining a normalization process in which patients negotiate their shifting realities (Core Category). Nature functioned as a support structure and nurtured patients' inner and outer capacities to respond and connect more effectively (Theme A). Once enabled and comforted, patients could engage survival and reconstructive maneuvers and explore the consequences of cancer (Theme B). A dynamic relationship was evident between moving away while, simultaneously, advancing towards the cancer reality in order to accept a shifting normality. From a place of comfort and safety, patients felt supported to deal differently and more creatively with the threat and demands of cancer diagnosis, treatment and outlook. New understanding about nature's role in cancer patients' lives calls attention to recognizing additional forms of psychosocial care that encourage patients' own coping and creative processes to deal with their strain and, in some cases, reconstruct everyday lives. Further research is required to determine how nature opportunities can be feasibly delivered in the cancer care setting.


Variation in OPRM1 moderates the effect of desire to drink on subsequent drinking and its attenuation by naltrexone treatment.
To evaluate the role of the functional Asn40Asp polymorphism in the mu-opioid receptor gene on drinking behavior and naltrexone's ability to attenuate drinking, we used a daily diary method in a 12-week, randomized clinical trial of naltrexone to reduce drinking. Participants (n = 158 problem drinkers) were assigned to receive either daily or targeted naltrexone 50 mg (n = 81) or matching placebo (n = 77). Patients reported by telephone each evening their current desire to drink and their drinking during the previous night and during the reporting day. We examined genotype, medication, desire to drink and their interactions as predictors of nighttime drinks consumed, controlling for drinking earlier in the day. Asp40 carriers showed a stronger positive association between evening desire (deviations from their mean levels) and later night drinking levels than Asn40 homozygotes (P = 0.019). The desire × genotype × medication condition interaction was also significant (P = 0.009), with a significant desire × genotype interaction for the placebo group (P = 0.001) but not for the naltrexone group (P = 0.74). In summary, when the evening level of desire to drink was relatively high, Asp40 allele carriers were at greater risk than Asn40 homozygotes to drink more, which was attenuated by naltrexone. Although average measures across the study were not informative, daily reports helped to demonstrate the moderating effects of genetic variation on the relation between desire to drink and alcohol consumption and the effects of naltrexone on that phenotype.


Genetic differentiation and estimation of gene flow from F-statistics under isolation by distance.
I reexamine the use of isolation by distance models as a basis for the estimation of demographic parameters from measures of population subdivision. To that aim, I first provide results for values of F-statistics in one-dimensional models and coalescence times in two-dimensional models, and make more precise earlier results for F-statistics in two-dimensional models and coalescence times in one-dimensional models. Based on these results, I propose a method of data analysis involving the regression of FST/(1-FST) estimates for pairs of subpopulations on geographic distance for populations along linear habitats or logarithm of distance for populations in two-dimensional habitats. This regression provides in principle an estimate of the product of population density and second moment of parental axial distance. In two cases where comparison to direct estimates is possible, the method proposed here is more satisfactory than previous indirect methods.


Precise Patterning of Organic Single Crystals via Capillary-Assisted Alternating-Electric Field.
Owing to the extraordinary properties, organic micro/nanocrystals are important building blocks for future low-cost and high-performance organic electronic devices. However, integrated device application of the organic micro/nanocrystals is hampered by the difficulty in high-throughput, high-precision patterning of the micro/nanocrystals. In this study, the authors demonstrate, for the first time, a facile capillary-assisted alternating-electric field method for the large-scale assembling and patterning of both 0D and 1D organic crystals. These crystals can be precisely patterned at the photolithography defined holes/channels at the substrate with the yield up to 95% in 1 mm2 . The mechanism of assembly kinetics is systematically studied by the electric field distribution simulation and experimental investigations. By using the strategy, various organic micro/nanocrystal patterns are obtained by simply altering the geometries of the photoresist patterns on substrates. Moreover, ultraviolet photodetectors based on the patterned Alq3 micro/nanocrystals exhibit visible-blind photoresponse with high sensitivity as well as excellent stability and reproducibility. This work paves the way toward high-integration, high-performance organic electronic, and optoelectronic devices from the organic micro/nanocrystals.


Cycle day, estrogen level, and lead follicle size: analysis of 27,790 in vitro fertilization cycles to determine optimal start criteria for gonadotropin-releasing hormone antagonist.
To determine the optimal criteria at which to start GnRH antagonists during controlled ovarian hyperstimulation (COH) for in vitro fertilization (IVF). Retrospective clinical cohort. IVF clinics. Women undergoing fresh autologous IVF using GnRH antagonist for ovulation suppression during COH. Measurement of lead follicle size, E2 level, and cycle day of stimulation on day of antagonist initiation. Clinical pregnancy rate (PR). The highest clinical PR was achieved when the antagonist was started when a lead follicle reached 14-15.9 mm in size (mean clinical PR 21.3; 95% confidence interval [CI] 19.3, 23.6) on cycle day 6 (mean clinical PR 22.2; 95% CI 17, 28.4), or when the E2 level was between 500 and 599 pg/mL (mean clinical PR 25.4; 95% CI 19.5, 32.4). Starting antagonists when the E2 level was <300 or >1,100 pg/mL reduced the odds of clinical pregnancy by 40% (odds ratio 0.60, 95% CI 0.5, 0.7). Cycle day, E2 level, and follicle size at time of antagonist start are all independent predictors of a clinical pregnancy after IVF. Initiating antagonists when the E2 level is extremely low (<300 pg/mL) or extremely high (>1,100 pg/mL) significantly reduces the odds of pregnancy.


Synthesis of protein in the pancreas. II. The role of ribonucleoprotein in protein synthesis.
1. The ribonucleoprotein of the microsome fraction which sediments at 40,000 R.P.M. as a pellet (and which is referred to as the pellet material) has been studied with reference to its role in protein synthesis in the pancreas. 2. In pellet material nucleic acid and protein form a definite complex as shown by its electrophoretic behavior and unchanging composition under various conditions. 3. Protein of pellet material is not especially rich in the diamino acids. 4. Evidence is brought forward indicating that the protein component of pellet material takes part in the general process of protein synthesis in the cell. (a) The well known correlation between quantity of RNA and rate of protein synthesis in a tissue implicates the protein of the pellet material, for most of the RNA in the pancreas and other tissues is in this material. (b) Uptake of isotopically labelled glycine by the pellet material, confirming results of previous workers, is for short periods greater than in other protein fractions. (c) Comparing the pellet materials of pancreas, liver, and kidney-three tissues with vastly different rates of protein synthesis, in the sequence given-there is a correlation between the quantity of RNA in the pellet and the rate of protein synthesis in the tissue; a similar correlation between quantity of RNA in the pellet material and rate of N(15)-glycine uptake by the protein component of the pellet; and finally, the level of uptake by total protein varies with the tissue and is related to the uptake of N(15)-glycine by protein of the pellet. 5. In the pancreas a distinction can be made between proteins synthesized for secretion and the nucleoprotein of the pellet (not found in the secretion) which, however, takes part in the synthetic process, as shown by the fact that the N(15) uptake by protein of the pellet is increased when the synthesis of digestive enzymes is stimulated by secretion. 6. The time course of N(15) uptake by proteins of the pancreas indicates that pellet protein serves as precursor material in the synthesis of the secretory proteins. 7. Rate of uptake of N(15)-glycine by the purines of RNA of the pellet material is not correlated with uptake by the protein. 8. The uptake of C(14)-alanine by an in vitro system of microsomes + mitochondria is impaired by preincubation of the microsomes with ribonuclease. This is direct experimental evidence for the dependence of protein synthesis upon the presence or intactness of ribonucleic acid in the microsomes.


Pediatric laparoscopic splenectomy using the lateral approach.
Laparoscopic splenectomy in children has been shown to be safe, to reduce postoperative pain and hospital stay, and to accelerate return to full activities. We describe our experience with a four-port "lateral" approach in 18 patients. Patients were placed in the lateral decubitus position and the table was flexed to separate the left subcostal margin and iliac crest. The camera port was inserted at the umbilicus and additional ports were placed in the epigastrium and left lower quadrant. After mobilization of the splenic flexure a port was inserted in the left flank below the 12th rib for elevation of the spleen. A 30 degrees laparoscope was used and the splenic vessels were controlled with an endo-GIA and/or clips. The spleens were placed in a bag, morcellated, and extracted through a port site. Eight females and 10 males with a median age of 12.5 years (5-17 years) and weight of 55.5 kg (17-124 kg) underwent splenectomy of idiopathic thrombocytopenia purpora (10), spherocytosis (6), elliptocytosis (1), and Hodgkin's disease (1). The median operating time was 160 min (90-300 min) and median blood loss was 105 ml (5-350 ml). Accessory spleens were removed in four cases. Three patients required extensions of a port site to remove large spleens which could not be placed in a bag. The sole complication was a transient pancreatitis with associated pleural effusion. The median postoperative hospital stay was 2 days (1-11 days) and time to full activities was 8 days (3-25 days). The lateral approach affords excellent visualization of the splenic vessels, pancreas, and accessory spleens. This approach is safe and reliable and is our preferred approach for laparoscopic splenectomy in children.


Biological and environmental factors affecting ultrasound-induced hemolysis in vitro: 2. Medium dissolved gas (pO2) content.
The data collected in this project supported the a priori hypothesis that the concentration of dissolved oxygen in whole human blood in vitro affected the extent of ultrasound (US)-induced hemolysis under conditions conducive to the occurrence of inertial cavitation. Aliquots of whole human blood in vitro with a relatively high O(2) level had statistically significantly more 1-MHz US-induced hemolysis than aliquots with a relatively low O(2) level in the presence of controlled gas nucleation (Albunex or ALX, supplementation), with US-induced hemolytic yields being substantially less at 2.2- and 3.5-MHz exposures or in the absence of ALX-supplementation at otherwise comparable acoustic pressures, pulse lengths and duty factors. Passive cavitation detection (pcd) measures indicated a linear relationship for hemolysis up to about 70% and pcd values (R(2) = 0.99).


Changes in economic outcomes before and after rural hospital closures in the United States: A difference-in-differences study.
The objective of this study is to assess changes in local economic outcomes before and after rural hospital closures. Rural hospital closures from January 1, 2005, to December 31, 2018, were obtained from the Sheps Center for Health Services Research. Economic outcomes from this same period were obtained from the Bureau of Labor Statistics, Bureau of Economic Analysis, Quarterly Workforce Indicators, U.S. Federal Reserve Economic Data, RAND Corporation state statistics database, U.S. Social Security Administration, and U.S. Census Bureau. Difference-in-differences study of 2094 rural counties. The primary exposure was county-level rural hospital closures. The primary outcomes were county-level unemployment rates; employment-population ratios; labor force participation-population ratios; per capita income; total jobs; health care sector jobs; disability program participation-population ratios; percent of the population with subprime credit scores; total filings for bankruptcies per 1000 population; and population size. A total of 104 rural counties experienced a hospital closure, compared to 1990 rural counties that did not. Rural hospital closures were associated with significant reductions in health care sector employment (-13.8%; 95% CI: -22%, -5.6%; p < 0.001), but not with changes in any other economic measure. For unemployment rates, employment-population ratios, per capita income, disability program participation-population ratios, and total jobs, we found evidence of adverse trends preceding hospital closures. Findings were robust to adjusting for county-specific time trends, specifying exposure at the commuting zone-level, and using alternate definitions of rurality to define sample counties. With the exception of a decline in jobs within the health care sector, there was no association between rural hospital closures and county-level economic outcomes. Instead, economic conditions were already declining in counties experiencing closures compared to those that did not.


Health technology assessment to optimize health technology utilization: using implementation initiatives and monitoring processes.
The way in which a health technology is used in any particular health system depends on the decisions and actions of a variety of stakeholders, the local culture, and context. In 2009, the HTAi Policy Forum considered how health technology assessment (HTA) could be improved to optimize the use of technologies (in terms of uptake, change in use, or disinvestment) in such complex systems. In scoping, it was agreed to focus on initiatives to implement evidence-based guidance and monitoring activities. A review identified systematic reviews of implementation initiatives and monitoring activities. A two-day deliberative workshop was held to discuss key papers, members' experiences, and collectively address key questions. This consensus paper was developed by email and finalized at a postworkshop meeting. Evidence suggests that the impact and use of HTA could be increased by ensuring timely delivery of relevant reports to clearly determined policy receptor (decision-making) points. To achieve this, the breadth of assessment, implementation initiatives such as incentives and targeted, intelligent dissemination of HTA result, needs to be considered. HTA stakeholders undertake a variety of monitoring activities, which could inform optimal use of a technology. However, the quality of these data varies and is often not submitted to an HTA. Monitoring data should be sufficiently robust so that they can be used in HTA to inform optimal use of technology. Evidence-based implementation initiatives should be developed for HTA, to better inform decision makers at all levels in a health system about the optimal use of technology.


Coinfection with Chlamydia trachomatis, Neisseria gonorrhoeae and Trichomonas vaginalis: a cross-sectional analysis of positivity and risk factors in remote Australian Aboriginal communities.
To determine the co-occurrence and epidemiological relationships of Chlamydia trachomatis (CT), Neisseria gonorrhoeae (NG) and Trichomonas vaginalis (TV) in a high-prevalence setting in Australia. In the context of a cluster randomised trial in 68 remote Aboriginal communities, we obtained laboratory reports on simultaneous testing for CT, NG and TV by nucleic acid amplification tests in individuals aged ≥16 years and examined relationships between age and sex and the coinfection positivity. ORs were used to determine which infections were more likely to co-occur by demographic category. Of 13 480 patients (median age: 30 years; men: 37%) tested for all three infections during the study period, 33.3% of women and 21.3% of men had at least one of them, highest in patients aged 16-19 years (48.9% in women, 33.4% in men). The most frequent combination was CT/NG (2.0% of women, 4.1% of men), and 1.8% of women and 0.5% of men had all three. In all co-combinations, coinfection positivity was highest in patients aged 16-19 years. CT and NG were highly predictive of each other's presence, and TV was associated with each of the other two infections, but much more so with NG than CT, and its associations were much stronger in women than in men. In this remote high-prevalence area, nearly half the patients aged 16-19 years had one or more sexually transmitted infections. CT and NG were more common dual infections. TV was more strongly associated with NG coinfections than with CT. These findings confirm the need for increased simultaneous screening for CT, NG and TV, and enhanced control strategies. Australian and New Zealand Clinical Trials Registry ACTRN12610000358044.


A Novel Selective Medium for Simultaneous Enrichment of Shiga Toxin-Producing Escherichia coli and Salmonella in Ground Beef.
Microbiological analysis of ground beef for contamination by both Salmonella and Shiga toxin-producing Escherichia coli (STEC) is performed by the U.S. Department of Agriculture, Food Safety Inspection Service (FSIS), as part of its Performance Standards Verification Testing program. FSIS has established a zero tolerance for STEC serotype O157:H7 and serogroups O26, O45, O103, O111, O121, and O145 because they are regarded as adulterants. The detection and isolation of these specific serogroups presents a technical challenge necessitating time-consuming and costly laboratory procedures that often exceed the technical capabilities of many small internal and reference laboratories. We describe here a method using a novel STEC and Salmonella selective (SSS) broth that allows for simultaneous selective enrichment of STEC and Salmonella sp., providing isolation and detection from the same broth. The method only involves direct plating from beef enrichments to detect suspect isolates that can be easily confirmed by using immunoassays or PCR, rendering the isolation simpler and less costly than the current described methods. In a side-by-side comparison with modified tryptic soy broth (mTSB), the use of SSS broth resulted in primarily isolating STEC and Salmonella sp., while substantially suppressing the growth of other gram-negative Enterobacteriacae by 90%. Significantly more (χ2 < 3.84) samples containing E. coli O157:H7 and STEC O26, O111, O121, and O145 and a nondifferent (χ2 > 3.84) number of samples containing STEC O103 and O45 were identified when enriching in SSS broth. Coenrichment using six different Salmonella serovars showed numerically greater but not significant (χ2 < 3.84) positive samples by using SSS broth compared with mTSB for a majority of serotypes.


Maternal Zika Virus Disease Severity, Virus Load, Prior Dengue Antibodies, and Their Relationship to Birth Outcomes.
Congenital Zika virus (ZIKV) syndrome is a newly identified condition resulting from infection during pregnancy. We analyzed outcome data from a mother-infant cohort in Rio de Janeiro in order to assess whether clinical severity of maternal ZIKV infection was associated with maternal virus load, prior dengue antibodies, or abnormal pregnancy/infant outcomes. A clinical severity assessment tool was developed based on duration of fever, severity of rash, multisystem involvement, and duration of symptoms during ZIKV infection. ZIKV-RNA load was quantified by polymerase chain reaction (PCR) cycles in blood/ urine. Dengue immunoglobulin G (IgG) antibodies were measured at baseline. Adverse outcomes were defined as fetal loss or a live infant with grossly abnormal clinical or brain imaging findings. Regression models were used to study potential associations. 131 ZIKV-PCR positive pregnant women were scored for clinical disease severity, 6 (4.6%) had mild disease, 98 (74.8%) had moderate disease, and 27 (20.6%) severe manifestations of ZIKV infection. There were 58 (46.4%) abnormal outcomes with 9 fetal losses (7.2%) in 125 pregnancies. No associations were found between: disease severity and abnormal outcomes (P = .961; odds ratio [OR]: 1.00; 95% confidence interval [CI]: 0.796-1.270); disease severity and viral load (P = .994); viral load and adverse outcomes (P = .667; OR: 1.02; 95% CI: 0.922-1.135); or existence of prior dengue antibodies (88% subjects) with severity score, ZIKV-RNA load or adverse outcomes (P = .667; OR: 0.78; 95% CI: 0.255-2.397). Congenital ZIKV syndrome does not appear to be associated with maternal disease severity, ZIKV-RNA load at time of infection or existence of prior dengue antibodies.


Cognitive factors modulate activity within the human subthalamic nucleus during voluntary movement in Parkinson's disease.
Movement is accompanied by changes in the degree to which neurons in corticobasal ganglia loops synchronize their activity within discrete frequency ranges. Although two principal frequency bands--beta (15-30 Hz) and gamma (60-90 Hz)--have been implicated in motor control, the precise functional correlates of their activities remain unclear. Local field potential (LFP) recordings in humans with Parkinson's disease undergoing surgery for deep brain stimulation to the subthalamic nucleus (STN) indicate that spectral changes both anticipate movement and occur perimovement. The extent to which such changes are modulated by cognitive factors involved in making a correct response seems critical in characterizing the functional associations of these oscillations. Accordingly, by recording LFP activity from the STN in parkinsonian patients, we demonstrate that perimovement beta and gamma reactivity is modulated by task complexity in a dopamine-dependent manner, despite the dynamics of the movement remaining unchanged. In contrast, spectral changes occurring in anticipation of future movement were limited to the beta band and, although modulated by dopaminergic therapy, were not modulated by task complexity. Our findings suggest two dopamine-dependent processes indexed by spectral changes in the STN: (1) an anticipatory activity reflected in the beta band that signals the likelihood of future action but does not proactively change with the cognitive demands of the potential response, and (2) perimovement activity that involves reciprocal beta and gamma band changes and is not exclusively related to explicit motor processing. Rather perimovement activity can also vary with, and may reflect, the cognitive complexity of the task.


Production systems of Creole goat and their implications for a breeding programme.
The Creole goat is a local meat breed well adapted to the tropical environment of Guadeloupe, a French island in the Caribbean. A survey of 47 goat farmers was conducted in May 2008 to describe the Guadeloupean goat farming systems. It was the preliminary step for the implementation of a breeding programme for Creole goats. Farmers had 31 does on average. A small number (4%) kept only Creole goats. Most of them (62%) had a mixed herd of Creole and crossbreds. One-third of them (34%) reared only crossbred goats. Farmers appreciate the rusticity and resistance of the Creole goat but consider its growth as too slow. The most desired traits for goat selection were conformation and growth for males (77% of the answers). These traits were also important for females (30% of the answers). Maternal qualities were also frequently cited (maternal behaviour 23%, reproduction 20% and milk production 17%). Disease resistance was not seen as an important trait (10% and 7% of the answers for bucks and does, respectively). A typology constituted of five groups of farmers was also created. Farmers of three groups were retained to participate at a selection programme. They kept Creole goats and have expressed a strong willingness to join a selection programme. The results of the survey suggest that a breeding programme should mostly focus on the Creole goat as a maternal breed. Real consideration should be given to disease resistance. The Creole goat has indeed a key role to play in the sustainability of local farming systems.


Structure of the gating domain of a Ca2+-activated K+ channel complexed with Ca2+/calmodulin.
Small-conductance Ca2+-activated K+ channels (SK channels) are independent of voltage and gated solely by intracellular Ca2+. These membrane channels are heteromeric complexes that comprise pore-forming alpha-subunits and the Ca2+-binding protein calmodulin (CaM). CaM binds to the SK channel through the CaM-binding domain (CaMBD), which is located in an intracellular region of the alpha-subunit immediately carboxy-terminal to the pore. Channel opening is triggered when Ca2+ binds the EF hands in the N-lobe of CaM. Here we report the 1.60 A crystal structure of the SK channel CaMBD/Ca2+/CaM complex. The CaMBD forms an elongated dimer with a CaM molecule bound at each end; each CaM wraps around three alpha-helices, two from one CaMBD subunit and one from the other. As only the CaM N-lobe has bound Ca2+, the structure provides a view of both calcium-dependent and -independent CaM/protein interactions. Together with biochemical data, the structure suggests a possible gating mechanism for the SK channel.


Binding partners for the COOH-terminal appendage domains of the GGAs and gamma-adaptin.
The adaptor appendage domains are believed to act as binding platforms for coated vesicle accessory proteins. Using glutathione S-transferase pulldowns from pig brain cytosol, we find three proteins that can bind to the appendage domains of both the AP-1 gamma subunit and the GGAs: gamma-synergin and two novel proteins, p56 and p200. p56 elicited better antibodies than p200 and was generally more tractable. Although p56 and gamma-synergin bind to both GGA and gamma appendages in vitro, immunofluorescence labeling of nocodazole-treated cells shows that p56 colocalizes with GGAs on TGN46-positive membranes, whereas gamma-synergin colocalizes with AP-1 primarily on a different membrane compartment. Furthermore, in AP-1-deficient cells, p56 remains membrane-associated whereas gamma-synergin becomes cytosolic. Thus, p56 and gamma-synergin show very strong preferences for GGAs and AP-1, respectively, in vivo. However, the GGA and gamma appendages share the same fold as determined by x-ray crystallography, and mutagenesis reveals that the same amino acids contribute to their binding sites. By overexpressing wild-type GGA and gamma appendage domains in cells, we can drive p56 and gamma-synergin, respectively, into the cytosol, suggesting a possible mechanism for selectively disrupting the two pathways.


Age-related cognitive decline in baboons: modeling the prodromal phase of Alzheimer's disease and related dementias.
The aging of brain cells and synaptic loss are the major underlying pathophysiological processes contributing to the progressive decline in cognitive functions and Alzheimer's disease. The difference in cognitive performances observed between adult and aged subjects across species highlights the decline of brain systems with age. The inflection point in age-related cognitive decline is important for our understanding of the pathophysiology of neurodegenerative diseases and for timing therapeutic interventions. Humans and nonhuman primates share many similarities including age-dependent changes in gene expression and decline in neural and immune functions. Given these evolutionary conserved organ systems, complex human-like behavioral and age-dependent changes may be modeled and monitored longitudinally in nonhuman primates. We integrated three clinically relevant outcome measures to investigate the effect of age on cognition, motor function and diurnal activity in aged baboons. We provide evidence of a naturally-occurring age-dependent precipitous decline in movement planning, in learning novel tasks, in simple discrimination and in motivation. These results suggest that baboons aged ~20 years (equivalent to ~60 year old humans) may offer a relevant model for the prodromal phase of Alzheimer's disease and related dementias to investigate mechanisms involved in the precipitous decline in cognitive functions and to develop early therapeutic interventions.


Efficacy of meaning-centered group psychotherapy for cancer survivors: a randomized controlled trial.
The aim of this study was to assess the efficacy of meaning-centered group psychotherapy for cancer survivors (MCGP-CS) to improve personal meaning, compared with supportive group psychotherapy (SGP) and care as usual (CAU). A total of 170 cancer survivors were randomly assigned to one of the three study arms: MCGP-CS (n = 57); SGP (n = 56); CAU (n = 57). The primary outcome measure was the Personal Meaning Profile (PMP; total score). Secondary outcome measures were subscales of the PMP, psychological well-being (Scales of Psychological Well-being; SPWB), post-traumatic growth (Posttraumatic Growth Inventory), Mental Adjustment to Cancer (MAC), optimism (Life Orientation Test-Revised), hopelessness (Beck's Hopelessness Scale), psychological distress (anxiety and depression, Hospital Anxiety and Depression Scale; HADS) and quality of life (European Organization for Research and Treatment of Cancer Quality of Life Questionnaire; EORTC QLQ-C30). Outcome measures were assessed before randomization, post-intervention, and after 3 and 6 months of follow-up (FU). Linear mixed model analyses (intention-to-treat) showed significant differences between MCGP-CS, SGP and CAU on the total PMP score, and on (sub)scales of the PMP, SPWB, MAC and HADS. Post-hoc analyses showed significantly stronger treatment effects of MCGP-CS compared with CAU on personal meaning (d = 0.81), goal-orientedness (d = 1.07), positive relations (d = 0.59), purpose in life (d = 0.69); fighting spirit (d = 0.61) (post-intervention) and helpless/hopeless (d = -0.87) (3 months FU); and distress (d = -0.6) and depression (d = -0.38) (6 months FU). Significantly stronger effects of MCGP-CS compared with SGP were found on personal growth (d = 0.57) (3 months FU) and environmental mastery (d = 0.66) (6 months FU). MCGP-CS is an effective intervention for cancer survivors to improve personal meaning, psychological well-being and mental adjustment to cancer in the short term, and to reduce psychological distress in the long run.


Thermal effects on red muscle contractile performance in deep-diving, large-bodied fishes.
Bigeye thresher sharks (Alopias superciliosus) and swordfish (Xiphias gladius) are large, pelagic fishes, which make long-duration, diurnal foraging dives from warm, surface waters (18-24 °C) to cold waters beneath the thermocline (5-10 °C). In bigeye thresher sharks, the subcutaneous position of the red, aerobic swimming muscles (RM) suggests that RM temperature mirrors ambient during dives (i.e., ectothermy). In swordfish, the RM is closer to the vertebrae and its associated with vascular counter-current heat exchangers that maintain RM temperature above ambient (i.e., RM endothermy). Here, we sought to determine how exposure to a wide range of ambient temperatures (8, 16, 24 °C) impacted peak power output and optimum cycle (i.e., tailbeat) frequency (0.25, 0.5, 1 Hz) in RM isolated from both species. Bigeye thresher shark RM did not produce substantial power at high cycle frequencies, even at high temperatures; but it did produce relatively high power at slow cycle frequencies regardless of temperature. Swordfish RM produced more power when operating at a combination of fast cycle frequencies and higher temperatures. This suggests that swordfish RM benefits considerably more from warming than bigeye thresher shark RM, while the RM of both species was able to produce power at cold temperatures and slow cycle frequencies. Despite different thermal strategies (i.e., ectothermy vs. RM endothermy), the ability of the RM to power sustained swimming during foraging-related search behaviors may contribute to the unique ability of these fishes to successfully exploit food resources in deep, cold water.


Corn-Based Distillers' Grains in Diets for Feedlot Cattle Are Associated with the Burden of Escherichia coli O157 in Feces.
Inclusion of distillers' grains (DGs) has been associated with increased prevalence of Escherichia coli O157:H7 in cattle housed in research settings. Our objective was to quantify the relationship between inclusion of DGs in commercial feedlot rations and the burden of E. coli O157. A convenience sample of 10 feedlots was enrolled based on DG use in finishing diets; 1 cohort included 5 feedlots in which DGs were greater than 15% of the dietary dry matter and the other cohort consisted of 5 feedlots at a concentration less than 8%. Sampling occurred at each feedlot on four occasions at ∼6-week intervals. At each feedlot visit, 4 pens of cattle within 3 weeks of slaughter were selected and 24 freshly voided fecal pats were sampled. Ten-gram samples were enriched in 90 mL of modified tryptic soy broth with novobiocin (20 mg/L) for 14 h at 42°C. Enrichments were subjected to immunomagnetic separation, plating onto chromogenic agar with novobiocin (5 mg/L) and potassium tellurite (2.5 mg/L), incubation for 18 h at 37°C, and latex agglutination of morphologically typical colonies. E. coli O157 was recovered from 16.7% of 3840 samples. Adjusted prevalence was 14.3% after controlling for within-feedlot and within-pen clustering. Prevalence during each sampling period was 19.9% (round 1), 21.0% (round 2), 14.1% (round 3), and 11.7% (round 4). Prevalence varied between cohorts, but this difference varied over time (p = 0.06). Among those with greater than 15% of the diet as DGs, prevalence was greater than those with less than 8% inclusion for all rounds of sampling (p < 0.01). Averaged across time, prevalence was 23.9% and 9.4% for those with greater than 15% and those with less than 8% of DGs, respectively. While observational, these data provide real-world support of reports of increased E. coli O157:H7 burden associated with DG use in cattle diets.


Methods and processes for the reanalysis of the NINDS tissue plasminogen activator for acute ischemic stroke treatment trial.
Treatment group imbalances in baseline stroke severity in the NINDS intravenous t-PA for acute stroke treatment trial led to controversy regarding the efficacy of tissue plasminogen activator (t-PA) in the treatment of acute ischemic stroke. Describe the steps used to independently re-evaluate this trial. NIH appointed an independent multidisciplinary committee that gained access to the original data. We undertook analyses of t-PA efficacy accounting for this imbalance, as well as analyses to identify subgroups that experienced additional harm or benefit from t-PA. Analyses of time from stroke onset to treatment (OTT), blood pressure, and intracerebral hemorrhage are given as illustrations. Despite subgroup imbalances in baseline stroke severity, when t-PA was administered to acute ischemic stroke patients according to study protocol, there was a statistically significant and clinically important benefit of t-PA treatment resulting in a higher likelihood of having a favorable clinical outcome at 3 months. Moreover, we were unable to identify subgroups of patients between which t-PA treatment effect differed, albeit these analyses had low power. These data failed to support the NINDS investigators' conclusion that effect of t-PA therapy diminished with increasing values of OTT within the protocol-specified 3 h time limit. In addition, the blood pressure measurements were highly variable and inconsistently determined so as to be too unreliable for inclusion in analysis. With new NIH requirements for data-sharing, the frequency of re-analysis of clinical trial data may increase substantially. This re-evaluation provides a blueprint for future re-evaluations of other trials. These best practices include re-analysis of the study data, after suitable replication, by an independent multidisciplinary committee, including a skilled statistical programmer analyst. Primary investigators should address significant errors determined in such re-analyses.


Orthostatic panic as a key Vietnamese reaction to traumatic events: the case of September 11, 2001.
This article discusses a culturally specific response to traumatic events: orthostatic panic attacks among Vietnamese refugees. We compared the rate and severity of orthostatic panic as well as the rates and severity of associated flashbacks a month before and a month after September 11, 2001. After that date, the rate and severity of orthostatic panic greatly increased, as did the rate and severity of associated flashbacks. The central role of orthostatic panic as a response to traumatic events is illustrated through a patient's vignette. An explanation of why September 11 so profoundly influenced this population is adduced, including an explanation of why it resulted in considerable worsening of orthostatic panic.


Peculiarities of cell composition and cell proliferation in different type atherosclerotic lesions in carotid and coronary arteries.
Increased cell proliferation in early atherosclerotic lesions is recognized as an essential event in atherogenesis but the levels of cell proliferation in the different stages of atherosclerotic plague formation in different types of human large arteries are still insufficiently studied. In the present work, we studied intima thickness and proliferation of newly "infiltrated" hematogenous and resident cells in atherosclerotic lesions of the carotid and coronary arteries and compared these parameters with those in the aorta, which we reported in an earlier publication (Orekhov et al. [8]). Analysis of intima thickness and proliferation in grossly unaffected intima and in different types of atherosclerotic lesions (initial lesions, fatty streaks, lipofibrous plaques, and fibrous plaques) revealed that, although there were similar tendencies in the change of the infiltration levels of hematogenous cells and proliferation in different types of arteries, there were significant quantitative differences between different types of arteries. Hematogenous cells in lipofibrous plaques of the coronary and carotid arteries were found to account for a third and almost for a half of the total cell population, respectively, while atherosclerotic lesions in the aorta, as shown by us previously, contain no more than 15% of hematogenous cells. This suggests that the contribution of hematogenous cells to the development of atherosclerosis in the carotid and coronary arteries appears to be more significant than in the aorta. Despite the differences in the numbers of accumulating hematogenous cells in the intima, a similar "bell-shaped" dependence of cell numbers on the lesion type, involved the following sequence: unaffected intima--initial lesions--fatty streaks--lipofibrous plaques--fibrous plaques, was detected in the coronary and carotid arteries. The visualization of PCNA-positive cells in atherosclerotic and unaffected zones of the coronary and carotid arteries revealed similar patterns of the distribution of proliferating cells. The maximum numbers of PCNA-positive resident cells were identified in lipofibrous plaques. The changes in the total cell numbers were found to be accompanied by the changes in the numbers of both proliferating resident cells and proliferating hematogenous cells. According to our knowledge, this is the first report that provides factual data about the similarities and differences in cell composition and proliferation between different types of large arteries in which the development of atherosclerosis is of crucial importance.


Proposed model of the relationship of risk information seeking and processing to the development of preventive behaviors.
We articulate a model that focuses on characteristics of individuals that might predispose them to seek and process information about health in different ways. Specifically, the model proposes that seven factors-(1) individual characteristics, (2) perceived hazard characteristics, (3) affective response to the risk, (4) felt social pressures to possess relevant information, (5) information sufficiency, (6) one's personal capacity to learn, (7) beliefs about the usefulness of information in various channels-will influence the extent to which a person will seek out this risk information in both routine and nonroutine channels and the extent to which he or she will spend time and effort analyzing the risk information critically. By adapting and synthesizing aspects of Eagly and Chaiken's Heuristic-Systematic Model and Ajzen's Theory of Planned Behavior, we also expect that people who engage in more effortful information seeking and processing are more likely to develop risk-related cognitions, attitudes, and behaviors that are more stable (i.e., less changeable or volatile) over time. Since most forms of health information campaigns attempt to get people to adopt habitual or lifestyle changes, factors leading to the stability or volatility of those behavioral changes are essential concerns.


Coordination of rapid cholinergic and dopaminergic signaling in striatum during spontaneous movement.
Interplay between dopaminergic and cholinergic neuromodulation in the striatum is crucial for movement control, with prominent models proposing pro-kinetic and anti-kinetic effects of dopamine and acetylcholine release, respectively. However, the natural, movement-related signals of striatum cholinergic neurons and their relationship to simultaneous variations in dopamine signaling are unknown. Here, functional optical recordings in mice were used to establish rapid cholinergic signals in dorsal striatum during spontaneous movements. Bursts across the cholinergic population occurred at transitions between movement states and were marked by widespread network synchronization which diminished during sustained locomotion. Simultaneous cholinergic and dopaminergic recordings revealed distinct but coordinated sub-second signals, suggesting a new model where cholinergic population synchrony signals rapid changes in movement states while dopamine signals the drive to enact or sustain those states.


Spatial organization of hominin activities at Gesher Benot Ya'aqov, Israel.
The spatial designation of discrete areas for different activities reflects formalized conceptualization of a living space. The results of spatial analyses of a Middle Pleistocene Acheulian archaeological horizon (about 750,000 years ago) at Gesher Benot Ya'aqov, Israel, indicate that hominins differentiated their activities (stone knapping, tool use, floral and faunal processing and consumption) across space. These were organized in two main areas, including multiple activities around a hearth. The diversity of human activities and the distinctive patterning with which they are organized implies advanced organizational skills of the Gesher Benot Ya'aqov hominins.


Individual variation in the propensity for prospective thought is associated with functional integration between visual and retrosplenial cortex.
It is well recognized that the default mode network (DMN) is involved in states of imagination, although the cognitive processes that this association reflects are not well understood. The DMN includes many regions that function as cortical "hubs", including the posterior cingulate/retrosplenial cortex, anterior temporal lobe and the hippocampus. This suggests that the role of the DMN in cognition may reflect a process of cortical integration. In the current study we tested whether functional connectivity from uni-modal regions of cortex into the DMN is linked to features of imaginative thought. We found that strong intrinsic communication between visual and retrosplenial cortex was correlated with the degree of social thoughts about the future. Using an independent dataset, we show that the same region of retrosplenial cortex is functionally coupled to regions of primary visual cortex as well as core regions that make up the DMN. Finally, we compared the functional connectivity of the retrosplenial cortex, with a region of medial prefrontal cortex implicated in the integration of information from regions of the temporal lobe associated with future thought in a prior study. This analysis shows that the retrosplenial cortex is preferentially coupled to medial occipital, temporal lobe regions and the angular gyrus, areas linked to episodic memory, scene construction and navigation. In contrast, the medial prefrontal cortex shows preferential connectivity with motor cortex and lateral temporal and prefrontal regions implicated in language, motor processes and working memory. Together these findings suggest that integrating neural information from visual cortex into retrosplenial cortex may be important for imagining the future and may do so by creating a mental scene in which prospective simulations play out. We speculate that the role of the DMN in imagination may emerge from its capacity to bind together distributed representations from across the cortex in a coherent manner.


Uncoupling of photoreceptor peripherin/rds fusogenic activity from biosynthesis, subunit assembly, and targeting: a potential mechanism for pathogenic effects.
Inherited defects in the RDS gene cause a multiplicity of progressive retinal diseases in humans. The gene product, peripherin/rds (P/rds), is a member of the tetraspanin protein family required for normal vertebrate photoreceptor outer segment (OS) architecture. Although its molecular function remains uncertain, P/rds has been suggested to catalyze membrane fusion events required for the OS renewal process. This study investigates the importance of two charged residues within a predicted C-terminal helical region for protein biosynthesis, localization, and interaction with model membranes. Targeted mutagenesis was utilized to neutralize charges at Glu(321) and Lys(324) individually and in combination to generate three mutant variants. Studies were conducted on variants expressed as 1) full-length P/rds in COS-1 cells, 2) glutathione S-transferase fusion proteins in Escherichia coli, and 3) membrane-associated green fluorescent protein fusion proteins in transgenic Xenopus laevis. None of the mutations affected biosynthesis of full-length P/rds in COS-1 cells as assessed by Western blotting, sedimentation velocity, and immunofluorescence microscopy. Although all mutations reside within a recently identified localization signal, none altered the ability of this region to direct OS targeting in transgenic X. laevis retinas. In contrast, individual or simultaneous neutralization of the charged amino acids Glu(321) and Lys(324) abolished the ability of the C-terminal domain to promote model membrane fusion as assayed by lipid mixing. These results demonstrate that, although overlapping, C-terminal determinants responsible for OS targeting and fusogenicity are separable and that fusogenic activity has been uncoupled from other protein properties. The observation that subunit assembly and OS targeting can both proceed normally in the absence of fusogenic activity suggests that properly assembled and targeted yet functionally altered proteins could potentially generate pathogenic effects within the vertebrate photoreceptor.


The outcome of physical symptoms with treatment of depression.
This study examined the prevalence, impact on health-related quality of life (HRQoL), and outcome of physical symptoms in depressed patients during 9 months of antidepressant therapy. Open-label, randomized, intention-to-treat trial with enrollment occurring April through November 1999. Thirty-seven primary care clinics within a research network. Five hundred seventy-three depressed patients started on one of three selective serotonin reuptake inhibitors (SSRIs) by their primary care physician and who completed a baseline interview. Patients were randomized to receive fluoxetine, paroxetine, or sertraline. Outcomes assessed included physical symptoms, depression, and multiple domains of HRQoL. Prevalence of physical symptoms was determined at baseline and after 1, 3, 6, and 9 months of treatment. Stepwise linear regression models were used to determine the independent effects of physical symptoms and depression on HRQoL domains. Of the 14 physical symptoms assessed, 13 were present in at least a third to half of the patients at baseline. Each symptom showed the greatest improvement during the initial month of treatment. In contrast, depression continued to show gradual improvement over a 9-month period. Physical symptoms had a predominant effect on pain (explaining 17% to 18% of the variance), physical functioning (13%), and overall health perceptions (13% to 15%). Depression had the greatest impact on mental (26% to 45%), social (14% to 32%), and work functioning (9% to 32%). Physical symptoms are prevalent in depressed patients and initially improve in the first month of SSRI treatment. Unlike depression, however, improvement in physical symptoms typically plateaus with minimal resolution in subsequent months.


Transient IL-10 receptor blockade can enhance CD8(+) T cell responses to a simian adenovirus-vectored HIV-1 conserved region immunogen.
Viral vector vaccines designed to elicit CD8(+) T cells in non-human primates exert potent control of immunodeficiency virus infections; however, similar approaches have been unsuccessful in humans. Adenoviral vectors elicit potent T cell responses but also induce production of immunosuppressive interleukin-10 (IL-10), which can limit the expansion of T cell responses. We investigated whether inhibiting IL-10 signaling prior to immunization with a candidate adenovirus vectored-HIV-1 vaccine, ChAdV63.HIVconsv, could modulate innate and adaptive immune responses in BALB/c mice. Transient IL-10 receptor blockade led to a modest but significant increase in the total magnitude CD8(+) T cell response to HIVconsv, but did not affect T cell responses to immunodominant epitopes. Anti-IL-10R-treated animals also exhibited greater expression of CD86 on CD11c(+) dendritic cells. Our data support further investigation and optimization of IL-10 blocking strategies to improve the immunogenicity of vaccines based on replication-defective adenoviruses.


Retrospective analysis of P-glycoprotein-mediated drug-drug interactions at the blood-brain barrier in humans.
To date, the in vitro-in vivo correlation (IVIVC) of P-glycoprotein (P-gp)-mediated drug-drug interaction (DDI) at the blood-brain barrier (BBB) in rats indicated that the cutoff value to significantly affect the brain penetration of digoxin was [I,unbound/Ki] of 1, where I,unbound is the unbound plasma concentration of P-gp inhibitors. On the basis of the IVIVC in rats, we speculated that clinically used P-gp inhibitors do not cause DDI at the human BBB, because none of the compounds studied was [I,unbound/Ki]>1 at therapeutic doses. Recently, positron emission tomography studies with P-gp substrates, such as [(11)C]verapamil, [(11)C]N-desmethyl loperamide, and [(11)C]loperamide, together with potent P-gp inhibitors, have indicated that increases in the influx rate constant for brain entry were observed in humans. Therefore, we aimed to retrospectively analyze the results of P-gp-mediated DDIs with in vitro P-gp inhibition assays and to confirm the appropriate cutoff value. In vitro P-gp inhibition assays using verapamil, N-desmethyl loperamide, and loperamide as P-gp probe substrates were performed in human multidrug resistance protein 1-expressing LLC-PK1 cells. The efflux ratios decreased in the presence of P-gp inhibitors, and the Ki of tariquidar was 10 nmol/L, regardless of probe substrates. Taking the in vitro Ki and unbound plasma concentrations in clinical DDI studies together, the criterion [I,unbound/Ki] of 1 was an appropriate cutoff limit to observe significant P-gp-mediated DDI at the BBB in humans. On the other hand, no significant DDI was observed in cases in which [I,unbound/Ki] was less than 0.1. This criterion was comparable to the previous IVIVC result in rats.


Dissociation between IFN-alpha-induced anti-viral and growth signaling pathways.
The ability of IFN-alpha to induce an anti-viral state in a wide variety of cell types as well as to inhibit cellular growth has long been appreciated. It is less clear, however, whether both these effects lie downstream of a common signaling pathway. In this study we have taken advantage of an atypical human myeloma cell line (KAS-6/1) displaying a dramatic proliferative response to IFN-alpha in an effort to resolve the signaling requirements for IFN-alpha-induced anti-viral and growth regulatory effects. Thus, we have analyzed the ability of IFN-alpha to induce a number of known receptor-initiated events in this cell line and have compared these responses with those exhibited by a cell lineage- and maturation stage-matched myeloma cell line (ANBL-6) that displays typical IFN-alpha responsiveness. Despite the widely contrasting effects of IFN-alpha on cellular proliferation, IFN-alpha was shown to be comparable in its ability to induce the expression of early response genes as well as induce resistance to viral infection in both cell lines. By contrast, the effects of IFN-alpha on the activation of mitogen-activated protein kinase (MAPK) were strikingly distinct. Finally, although inhibition of MEK and MAPK activation had no effect on the induction of the anti-viral response, it completely blocked IFN-alpha-stimulated proliferation of the KAS-6/1 cells. In summary, our analysis of the role of the MAPK and anti-viral signaling pathways using these two cell lines suggests that the anti-viral and growth regulatory effects of IFN-alpha display a differential requirement for activation of the MAPK pathway.


How Comprehensively Is Evidence-Based Practice Represented in Australian Health Professional Accreditation Documents? A Systematic Audit.
PHENONENON: In many developed countries, accreditation documents, which reflect the practice standards of health professions, form the basis for evaluation of education programs for meeting the requirements for registration. The 2005 Sicily statement proposed a 5-step model of training in evidence-based practice (ask, access, appraise, apply, and assess). A key recommendation was that evidence-based practice should be incorporated into entry-level health professional training and registration. No previous research has assessed the extent to which this has occurred. We undertook a systematic audit of the accreditation documents for the registered health professions in Australia. The 11 health professional disciplines included in the audit were medicine, nursing and midwifery, pharmacy, physiotherapy, dentistry, psychology, occupational therapy, optometry, podiatry, osteopathy, and chiropractic. Two investigators independently identified the occurrence of the term evidence that related to "evidence-based practice" and the occurrences of terms related to the 5 steps in the accreditation documents. Occurrence of the term evidence as it relates specifically to "evidence-based practice" ranged from 0 (pharmacy, dentistry and occupational therapy) to 8 (physiotherapy) in the accreditation documents. Overall, there were 77 occasions when terms relating to any of the 5 steps of evidence-based practice were used across all 11 accreditation documents. All 5 steps were included in the physiotherapy and psychology documents; 4 steps in medicine and optometry; 3 steps in pharmacy; 2 steps each in documents for chiropractic, osteopathy, and podiatry; and 1 step for nursing. There was no inclusion of terms relating to any of the 5 steps in the dentistry and occupational therapy documents. Insights: Terminology relating explicitly to evidence-based practice and to the 5 steps of evidence-based practice appears to be lacking in the accreditation documents for health professions registered in Australia. This is not necessarily reflective of the curricular content or quality, or dedication to evidence-based practice teaching. However, recognition and demand by accreditation bodies for skills in evidence-based practice may act as a driver for education providers to give greater priority to embedding this training in entry-level programs. Consequently, accreditation bodies are powerfully positioned to shape future directions, focus, and boundaries within and across professions. Future international audits of accreditation documents could provide insight into the global breadth of this phenomenon and contribute to closer scrutiny of the representation of evidence-based practice in future iterations of accreditation documents.


Associations of supermarket characteristics with weight status and body fat: a multilevel analysis of individuals within supermarkets (RECORD study).
Previous research on the influence of the food environment on weight status has often used impersonal measures of the food environment defined for residential neighborhoods, which ignore whether people actually use the food outlets near their residence. To assess whether supermarkets are relevant contexts for interventions, the present study explored between-residential neighborhood and between-supermarket variations in body mass index (BMI) and waist circumference (WC), and investigated associations between brands and characteristics of supermarkets and BMI or WC, after adjustment for individual and residential neighborhood characteristics. Participants in the RECORD Cohort Study (Paris Region, France, 2007-2008) were surveyed on the supermarket (brand and exact location) where they conducted their food shopping. Overall, 7 131 participants shopped in 1 097 different supermarkets. Cross-classified multilevel linear models were estimated for BMI and WC. Just 11.4% of participants shopped for food primarily within their residential neighborhood. After accounting for participants' residential neighborhood, people shopping in the same supermarket had a more comparable BMI and WC than participants shopping in different supermarkets. After adjustment for individual and residential neighborhood characteristics, participants shopping in specific supermarket brands, in hard discount supermarkets (especially if they had a low education), and in supermarkets whose catchment area comprised low educated residents had a higher BMI/WC. A public health strategy to reduce excess weight may be to intervene on specific supermarkets to change food purchasing behavior, as supermarkets are where dietary preferences are materialized into definite purchased foods.


Investigation of Drowning Deaths: A Practical Review.
Drowning, which typically involves a watery environment, remains a serious public health concern claiming an estimated 362 000 lives per year worldwide across all socioeconomic classifications and has remained under close observation by the World Health Organization and its signatories. A significant number of water-related deaths are attributed to accidental drowning, while a smaller but still significant number represent suicidal or homicidal drowning. Others involve a combination of drowning precipitated by injury, intoxication, or environmental extremes. Still others involve victims that die from injury, intoxication, or a natural disease entity of such significance as to preclude the drowning process, while near or in water. While there may be an initial presumption that all water-related deaths are accidental drownings, other possibilities must be considered in the investigation of these types of deaths, as drowning as a cause of death is a diagnosis based on the exclusion of other potential causes. The coordinated investigative efforts of multiple agencies and disciplines are required not only for the designation as drowning as the cause of death but also for death certification. The ongoing analysis and dissemination of data generated from all levels of investigation augment our understanding of the impact on public health and safety, guiding allocation of monetary and educational resources in an effort to prevent further mortality and disability.


Anxiety disorders: Psychiatric comorbidities and psychosocial stressors among adult outpatients.
Anxiety disorders are the most prevalent class of lifetime mental disorders according to South African research. However, little is known about the prevalence of factors that might complicate treatment among adults in a psychiatric outpatient setting. To explore the psychiatric comorbidities and psychosocial stressors among a population of adults treated for anxiety disorders at the outpatient unit of a tertiary psychiatric facility in Bloemfontein. In this retrospective cross-sectional study, clinical files of all mental healthcare users receiving treatment were reviewed to identify those with a current or previous diagnosis of one or more of the following anxiety disorders: generalised anxiety disorder (GAD), panic disorder, social anxiety disorder (SAD) and agoraphobia. Of the 650 available records, 103 (15.8%) included at least one anxiety disorder. Of those, 65.1% had GAD, 34.0% had panic disorder and 29.1% had SAD. Agoraphobia was diagnosed in 14.6% of patients almost exclusively as comorbid with panic disorder. Additional psychiatric disorders were present for 98.1% of patients and 36.9% had multiple anxiety disorders. The patients had a history of relational problems (64.1%), educational and occupational stressors (55.3%), abuse and neglect (28.2%), other problems related to the social environment (24.3%) and self-harm (23.3%). Clinical practice should take the high rates of comorbidity into account and the importance of integrated substance-related interventions in mental healthcare settings is clear. Diagnostic practices regarding agoraphobia without panic, and the comorbidity of anxiety and personality disorders should receive further attention. Clinicians should be aware of the potential impact of the frequently reported psychosocial stressors.


Would primary healthcare professionals prescribe a polypill to manage cardiovascular risk? A qualitative interview study.
A 'polypill' containing both blood pressure-lowering and cholesterol-lowering drugs could prevent up to 80% of cardiovascular disease events. Since little is known about the attitudes of primary healthcare professionals to use of such a pill for cardiovascular disease prevention, this study aimed to investigate opinions. Semistructured interviews were conducted with participants. A qualitative description approach was used to analyse and report the results. Participants were recruited from nine primary care practices in Birmingham. Sixteen healthcare professionals (11 primary care physicians and 5 practice nurses) were selected through purposive sampling to maximise variation of characteristics. Outcome measures for this study were: the attitude of healthcare professionals towards the use of a polypill for primary and secondary cardiovascular disease prevention; their views on monitoring the drug; and the factors influencing their willingness to prescribe the medication. Healthcare professionals expressed considerable concern over using a polypill for primary prevention for all people over a specific age, although there was greater acceptance of its use for secondary prevention. Regularly monitoring patients taking the polypill was deemed essential. Evidence of effectiveness, patient risk level and potential medicalisation were key determinants in willingness to prescribe such a pill. Primary healthcare professionals have significant concerns about the use of a polypill, particularly in the prevention of cardiovascular disease in people who are not regarded as being at 'high risk'. If a population-based polypill strategy is to be successfully implemented, healthcare professionals will need to be convinced of the potential benefits of a drug-based population approach to prevention.


Modulation of erythrocyte sensitivity to oxidative stress by transient hyperhomocysteinemia in healthy subjects and in patients with coronary artery disease.
The pathobiological mechanisms by which Hcy can promote atherothrombosis are not completely understood. Many observations suggest that oxidative consequences of hyperhomocysteinemia have a distinct role in the development of occlusive vascular disease. The aim of this work was to investigate whether sensitivity of erythrocytes to chemically induced oxidative stress in both healthy subjects and patients with clinically ascertained atherosclerosis was modified during the transient increase in homocysteine driven by methionine load. Erythrocyte sensitivity to oxidative stress during transient hyperhomocysteinemia was assessed by cumene hydroperoxide-induced alpha-tocopherol consumption before and after methionine load in 31 healthy subjects and 23 patients with coronary artery disease. Decreased sensitivity to oxidative challenge ("Type-1" response) after methionine load was more frequent in healthy subjects (35% vs 13% in patients), while increased sensitivity ("Type-2" response) was more frequent in patients (22% vs 6% in healthy subjects). No variation in sensitivity to oxidative challenge throughout the loading test ("Non-variant" response) was detected in either group (58% in healthy subjects and 65% in patients). The distribution of these responses was significantly different between healthy subjects and patients and independent of basal and post-load increase in homocysteine. Plasma lipoperoxides, erythrocyte alpha-tocopherol and glutathione content before methionine load were significantly different between patients and healthy subjects; however only the redox potential of the GSSG/GSH couple was significantly different in the different groups of response. The higher frequency of "Type-2" response in patients with respect to healthy subjects suggests that methionine load reveals individual factors that may contribute to the pathogenesis of atherosclerosis.


Approaches to Foster Transfer of Formal Principles: Which Route to Take?
Enabling learners to transfer knowledge about formal principles to new problems is a major aim of science and mathematics education, which, however, is notoriously difficult to reach. Previous research advocates different approaches of how to introduce principles to foster the transfer of knowledge about formal principles. One approach suggests teaching a generic formalism of the principles. Another approach suggests presenting (at least) two concrete cases instantiating the principle. A third approach suggests presenting a generic formalism accompanied by a case. As yet, though, empirical results regarding the transfer potential of these approaches are mixed and difficult to integrate as the three approaches have rarely been tested competitively. Furthermore, the approaches have been evaluated in relation to different control conditions, and they have been assessed using varying transfer measures. In the present experiment, we introduced undergraduates to the formal principles of propositional logic with the aim to systematically compare the transfer potential of the different approaches in relation to each other and to a common control condition by using various learning and transfer tasks. Results indicate that all approaches supported successful learning and transfer of the principles, but also caused systematic differences in the magnitude of transfer. Results indicate that the combination of a generic formalism with a case was surprisingly unsuccessful while learners who compared two cases outperformed the control condition. We discuss how the simultaneous assessment of the different approaches allows to more precisely capture the underlying learning mechanisms and to advance theory on how these mechanisms contribute to transfer performance.


Maternal Obesity Management Using Mobile Technology: A Feasibility Study to Evaluate a Text Messaging Based Complex Intervention during Pregnancy.
Maternal obesity and excessive gestational weight gain (GWG) are on the rise with negative impact on pregnancy and birth outcomes. Research into managing GWG using accessible technology is limited. The maternal obesity management using mobile technology (MOMTech) study aimed at evaluating the feasibility of text messaging based complex intervention designed to support obese women (BMI ≥ 30) with healthier lifestyles and limit GWG. Participants received two daily text messages, supported by four appointments with healthy lifestyle midwife, diet and activity goal setting, and self-monitoring diaries. The comparison group were obese mothers who declined to participate but consented for their routinely collected data to be used for comparison. Postnatal interviews and focus groups with participants and the comparison group explored the intervention's acceptability and suggested improvements. Fourteen women completed the study which did not allow statistical analyses. However, participants had lower mean GWG than the comparison group (6.65 kg versus 9.74 kg) and few (28% versus 50%) exceeded the Institute of Medicine's upper limit of 9 kg GWG for obese women. MOMTech was feasible within clinical setting and acceptable intervention to support women to limit GWG. Before further trials, slight modifications are planned to recruitment, text messages, and the logistics of consultation visits.


Chlamydia--host cell interactions: recent advances on bacterial entry and intracellular development.
Bacteria of the Chlamydiales order are very successful intracellular organisms that grow in human and animal cells, and even in amoebae. They fulfill several essential functions to enter their host cells, establish an intracellular environment favorable for their multiplication and exit the host cell. They multiply in a unique organelle called the inclusion, which is isolated from the endocytic but not the exocytic pathway. A combination of host cell factors and of proteins secreted by the bacteria, from within the inclusion, contribute to the establishment and development of this inclusion. Here we review recent data on the entry mechanisms and maturation of the inclusion.


Beta catenin-independent activation of MyoD in presomitic mesoderm requires PKC and depends on Pax3 transcriptional activity.
Early activation of myogenesis in the somite depends on signals from surrounding tissues. Canonical beta-catenin dependent Wnt signalling preferentially activates Myf5. We now show, in explant experiments with presomitic mesoderm, that the expression of another myogenic determination factor, MyoD, depends on non-canonical Wnt signalling, probably emanating from the dorsal ectoderm. Inhibitors of PKC block MyoD expression, indicating that the intracellular Wnt pathway depends on this kinase. In the absence of Myf5 and Mrf4, this activation is only minorily affected and we identify Pax3 as the transcriptional mediator responsible for MyoD expression. When embryos expressing a constitutively active form of Pax3, PAX3-FKHR, are used for these studies in the presence of PKC inhibitors, MyoD expression is not affected, suggesting that Wnt signalling acts on the transcriptional activity of Pax3.


Loss-coupled DFB nano-ridge laser monolithically grown on a standard 300-mm Si wafer.
We present a loss-coupled distributed feedback microlaser, monolithically grown on a standard 300-mm Si wafer using nano-ridge engineering. The cavity is formed by integrating a metallic grating on top of the nano-ridge. This allows forming a laser cavity without etching the III-V material, avoiding damaged interfaces and the associated carrier loss. Simulations, supported by experimental characterisation of the modal gain of the nano-ridge devices, predict an optimal duty cycle for the grating of ~0.4, providing a good trade-off between coupling strength and cavity loss for the lasing mode. The model was experimentally verified by characterising the lasing threshold and external efficiency of devices exhibiting gratings with varying duty cycle. The high modal gain and low threshold obtained prove the excellent quality of the epitaxial material. Furthermore, the low loss metal grating might provide a future route to electrical injection and efficient heat dissipation of these nanoscale devices.


Dementia in Africa: Current evidence, knowledge gaps, and future directions.
In tandem with the ever-increasing aging population in low and middle-income countries, the burden of dementia is rising on the African continent. Dementia prevalence varies from 2.3% to 20.0% and incidence rates are 13.3 per 1000 person-years with increasing mortality in parts of rapidly transforming Africa. Differences in nutrition, cardiovascular factors, comorbidities, infections, mortality, and detection likely contribute to lower incidence. Alzheimer's disease, vascular dementia, and human immunodeficiency virus/acquired immunodeficiency syndrome-associated neurocognitive disorders are the most common dementia subtypes. Comprehensive longitudinal studies with robust methodology and regional coverage would provide more reliable information. The apolipoprotein E (APOE) ε4 allele is most studied but has shown differential effects within African ancestry compared to Caucasian. More candidate gene and genome-wide association studies are needed to relate to dementia phenotypes. Validated culture-sensitive cognitive tools not influenced by education and language differences are critically needed for implementation across multidisciplinary groupings such as the proposed African Dementia Consortium.


A novel PKC-regulated mechanism controls CD44 ezrin association and directional cell motility.
The dynamic assembly and disassembly of membrane cytoskeleton junctional complexes is critical in cell migration. Here we describe a novel phosphorylation mechanism that regulates the hyaluronan receptor CD44. In resting cells, CD44 is constitutively phosphorylated at a single serine residue, Ser325. After protein kinase C is activated, a switch in phosphorylation results in CD44 being phosphorylated solely at an alternative residue, Ser291. Using fluorescence resonance energy transfer (FRET) monitored by fluorescence lifetime imaging microscopy (FLIM) and chemotaxis assays we show that phosphorylation of Ser291 modulates the interaction between CD44 and the cytoskeletal linker protein ezrin in vivo, and that this phosphorylation is critical for CD44-dependent directional cell motility.


Setting standards for severity of common symptoms in oncology using the PROMIS item banks and expert judgment.
Although the use of patient-reported outcome measures (PROs) has increased markedly, clinical interpretation of scores remains lacking. We developed a method to identify clinical severity thresholds for pain, fatigue, depression, and anxiety in people with cancer. Using available Patient-Reported Outcomes Measurement Information System (PROMIS) item bank response data collected on 840 cancer patients, symptom vignettes across a range of symptom severity were developed and placed on index cards. Cards represented symptom severity at five-point intervals differences on the T score metric [mean = 50; standard deviation (SD) = 10]. Symptom vignettes for each symptom were anchored on these standardized scores at 0.5 SD increments across the full range of severity. Clinical experts, blind to the PROMIS score associated with each vignette, rank-ordered the vignettes by severity, then arrived at consensus regarding which two vignettes were at the upper and lower boundaries of normal and mildly symptomatic for each symptom. The procedure was repeated to identify cut scores separating mildly from moderately symptomatic, and moderately from severely symptomatic scores. Clinician severity rankings were then compared to the T scores upon which the vignettes were based. For each of the targeted PROs, the severity rankings reached by clinician consensus perfectly matched the numerical rankings of their associated T scores. Across all symptoms, the thresholds (cut scores) identified to differentiate normal from mildly symptomatic were near a T score of 50. Cut scores differentiating mildly from moderately symptomatic were at or near 60, and those separating moderately from severely symptomatic were at or near 70. The study results provide empirically generated PROMIS T score thresholds that differentiate levels of symptom severity for pain interference, fatigue, anxiety, and depression. The convergence of clinical judgment with self-reported patient severity scores supports the validity of this methodology to derive clinically relevant symptom severity levels for PROMIS symptom measures in other settings.


Antiviral activity of Australian tea tree oil and eucalyptus oil against herpes simplex virus in cell culture.
The antiviral effect of Australian tea tree oil (TTO) and eucalyptus oil (EUO) against herpes simplex virus was examined. Cytotoxicity of TTO and EUO was evaluated in a standard neutral red dye uptake assay. Toxicity of TTO and EUO was moderate for RC-37 cells and approached 50% (TC50) at concentrations of 0.006% and 0.03%, respectively. Antiviral activity of TTO and EUO against herpes simplex virus type 1 (HSV-1) and herpes simplex virus type 2 (HSV-2) was tested in vitro on RC-37 cells using a plaque reduction assay. The 50% inhibitory concentration (IC50) of TTO for herpes simplex virus plaque formation was 0.0009% and 0.0008% and the IC50 of EUO was determined at 0.009% and 0.008% for HSV-1 and HSV-2, respectively. Australian tea tree oil exhibited high levels of virucidal activity against HSV-1 and HSV-2 in viral suspension tests. At noncytotoxic concentrations of TTO plaque formation was reduced by 98.2% and 93.0% for HSV-1 and HSV-2, respectively. Noncytotoxic concentrations of EUO reduced virus titers by 57.9% for HSV-1 and 75.4% for HSV-2. Virus titers were reduced significantly with TTO, whereas EUO exhibited distinct but less antiviral activity. In order to determine the mode of antiviral action of both essential oils, either cells were pretreated before viral infection or viruses were incubated with TTO or EUO before infection, during adsorption or after penetration into the host cells. Plaque formation was clearly reduced, when herpes simplex virus was pretreated with the essential oils prior to adsorption. These results indicate that TTO and EUO affect the virus before or during adsorption, but not after penetration into the host cell. Thus TTO and EUO are capable to exert a direct antiviral effect on HSV. Although the active antiherpes components of Australian tea tree and eucalyptus oil are not yet known, their possible application as antiviral agents in recurrent herpes infection is promising.


Targeted drug delivery by in vivo coupling to endogenous albumin: an albumin-binding prodrug of methotrexate (MTX) is better than MTX in the treatment of murine collagen-induced arthritis.
To examine the effect of an albumin-binding prodrug of methotrexate (MTX) in the treatment of murine collagen-induced arthritis (CIA). The prodrug AWO54 with the formula EMC-d-Ala-Phe-Lys-Lys-MTX binds selectively to the cysteine-34 position of endogenous albumin, which acts as a macromolecular drug carrier for MTX to the site of inflammation. The CIA model was used to evaluate the anti-arthritic effect of the compound after intravenous application. The albumin-bound form of AWO54 was efficiently cleaved by cathepsin B and plasmin, two proteases that are overexpressed in rheumatoid arthritis, and release a MTX lysine derivative. AWO54 suppressed CIA in a dose-dependent manner and was significantly better than MTX. To obtain a similar effect only about 20% of the MTX-equivalent dose of AWO54 had to be given. The efficacy of the drug was tested in two different stages of CIA: while both, MTX and AWO54 inhibited arthritis in an early stage of the disease, in a later stage only AWO54 showed a significant inhibitory effect in comparison with control. Targeted drug delivery by in vivo coupling of a prodrug of MTX to endogenous albumin is better than MTX in the treatment of CIA.


Insufficient Reporting of Factors Associated With Exercise Referral Scheme Uptake, Attendance, and Adherence: A Systematic Review of Reviews.
Exercise referral schemes (ERS) are prescribed programs to tackle physical inactivity and associated noncommunicable disease. Inconsistencies in reporting, recording, and delivering ERS make it challenging to identify what works, why, and for whom. Preferred Reporting Items for Systematic Reviews and Meta-Analyses guided this narrative review of reviews. Electronic databases were searched for systematic reviews of ERS. Inclusion criteria and quality assessed through A Measurement Tool to Assess Systematic Reviews (AMSTAR). Data on uptake, attendance, and adherence were extracted. Eleven reviews met inclusion criteria. AMSTAR quality was medium. Uptake ranged between 35% and 81%. Groups more likely to take up ERS included (1) females and (2) older adults. Attendance ranged from 12% to 49%. Men were more likely to attend ERS. Effect of medical diagnosis upon uptake and attendance was inconsistent. Exercises prescribed were unreported; therefore, adherence to exercise prescriptions was unreported. The influence of theoretically informed approaches on uptake, attendance, and adherence was generally lacking; however, self-determination, peer support, and supervision were reported as influencing attendance. There was insufficient reporting across studies about uptake, attendance, and adherence. Complex interventions such as ERS require consistent definitions, recording, and reporting of these key facets, but this is not evident from the existing literature.


Plasmid persistence: costs, benefits, and the plasmid paradox.
Plasmids are extrachromosomal DNA elements that can be found throughout bacteria, as well as in other domains of life. Nonetheless, the evolutionary processes underlying the persistence of plasmids are incompletely understood. Bacterial plasmids may encode genes for traits that are sometimes beneficial to their hosts, such as antimicrobial resistance, virulence, heavy metal tolerance, and the catabolism of unique nutrient sources. In the absence of selection for these traits, however, plasmids generally impose a fitness cost on their hosts. As such, plasmid persistence presents a conundrum: models predict that costly plasmids will be lost over time or that beneficial plasmid genes will be integrated into the host genome. However, laboratory and comparative studies have shown that plasmids can persist for long periods, even in the absence of positive selection. Several hypotheses have been proposed to explain plasmid persistence, including host-plasmid co-adaptation, plasmid hitchhiking, cross-ecotype transfer, and high plasmid transfer rates, but there is no clear evidence that any one model adequately resolves the plasmid paradox.