The Concentration of Opioid Prescriptions by Providers and Among Patients in the Oregon Medicaid Program.
This study examined the distribution of opioid prescribing across providers and patients and the extent to which concentrated distribution predicts opioid misuse. Using 2013 Oregon Medicaid claims and the National Provider Identifier Registry, this study identified patients who filled at least one opioid prescription and providers who prescribed opioids for those patients (N=61,477 Medicaid beneficiaries). This study examined the distribution of opioid prescriptions by provider and patient, the extent to which high-volume opioid use was associated with potential opioid misuse, and how this association changed when patients received opioids from providers in the top decile of morphine-equivalent doses (MEQ) prescribed in 2013. This study used four indicators of opioid misuse: doctor and pharmacy shopping for opioid prescriptions, opioid prescription overlap, and opioid and benzodiazepine prescription overlap. Opioid use and prescriptions were heavily concentrated among the top 10% of opioid users and prescribers. Those high-volume opioid users and prescribers accounted for, respectively, 83.2% and 80.8% in MEQ of entire opioids prescribed. Patients' increasing use of opioids (by MEQ) was associated with most measures of opioid misuse. Patients receiving opioids from high-volume prescribers had a higher probability of opioid prescription overlap and opioid and benzodiazepine prescription overlap compared with other patients, but the difference was significant only among patients who received high doses of opioids, and the size of the difference was modest. Whereas current policies emphasize reducing opioid prescriptions across all patients and providers, study results suggest that focusing policies on high-volume opioid users and prescribers may be more beneficial.


Induction of apoptosis of bladder cancer cells by zinc-citrate compound.
Zinc is one of the trace minerals in the body and is known to have an anticancer effect by inducing apoptosis in prostate cancer. We aimed to investigate the antiproliferative effects of a zinc-citrate compound in bladder cancer. A bladder cancer cell line (MBT-2) was treated with a zinc-citrate compound at different time intervals and concentrations. Mitochondrial (m)-aconitase activity was determined by use of the aconitase assay. DNA laddering analysis was performed to investigate apoptosis of MBT-2 cells. The molecular mechanism of apoptosis was investigated by Western blot analysis of p53, p21(waf1), Bcl-2, Bcl-xL, and Bax and also by caspase-3 activity analysis. Treatment with the zinc-citrate compound resulted in a time- and dose-dependent decrease in cell number of MBT-2 cells. M-aconitase activity was significantly decreased. DNA laddering analysis indicated apoptosis of MBT-2 cells. The zinc-citrate compound increased the expression of p21(waf1) and p53 and reduced the expression of Bcl-2 and Bcl-xL proteins but induced expression of Bax protein. The zinc-citrate compound induced apoptosis of MBT-2 cells by activation of the caspase-3 pathway. We have shown that a zinc-citrate compound induces apoptotic cell death in a bladder cancer cell line, MBT-2, by caspase-3 activation through up-regulation of apoptotic proteins and down-regulation of antiapoptotic proteins.


Assessing historical fish community composition using surveys, historical collection data, and species distribution models.
Accurate establishment of baseline conditions is critical to successful management and habitat restoration. We demonstrate the ability to robustly estimate historical fish community composition and assess the current status of the urbanized Barton Creek watershed in central Texas, U.S.A. Fish species were surveyed in 2008 and the resulting data compared to three sources of fish occurrence information: (i) historical records from a museum specimen database and literature searches; (ii) a nearly identical survey conducted 15 years earlier; and (iii) a modeled historical community constructed with species distribution models (SDMs). This holistic approach, and especially the application of SDMs, allowed us to discover that the fish community in Barton Creek was more diverse than the historical data and survey methods alone indicated. Sixteen native species with high modeled probability of occurrence within the watershed were not found in the 2008 survey, seven of these were not found in either survey or in any of the historical collection records. Our approach allowed us to more rigorously establish the true baseline for the pre-development fish fauna and then to more accurately assess trends and develop hypotheses regarding factors driving current fish community composition to better inform management decisions and future restoration efforts. Smaller, urbanized freshwater systems, like Barton Creek, typically have a relatively poor historical biodiversity inventory coupled with long histories of alteration, and thus there is a propensity for land managers and researchers to apply inaccurate baseline standards. Our methods provide a way around that limitation by using SDMs derived from larger and richer biodiversity databases of a broader geographic scope. Broadly applied, we propose that this technique has potential to overcome limitations of popular bioassessment metrics (e.g., IBI) to become a versatile and robust management tool for determining status of freshwater biotic communities.


Sunscreens as a source of hydrogen peroxide production in coastal waters.
Sunscreens have been shown to give the most effective protection for human skin from ultraviolet (UV) radiation. Chemicals from sunscreens (i.e., UV filters) accumulate in the sea and have toxic effects on marine organisms. In this report, we demonstrate that photoexcitation of inorganic UV filters (i.e., TiO2 and ZnO nanoparticles) under solar radiation produces significant amounts of hydrogen peroxide (H2O2), a strong oxidizing agent that generates high levels of stress on marine phytoplankton. Our results indicate that the inorganic oxide nanoparticle content in 1 g of commercial sunscreen produces rates of H2O2 in seawater of up to 463 nM/h, directly affecting the growth of phytoplankton. Conservative estimates for a Mediterranean beach reveal that tourism activities during a summer day may release on the order of 4 kg of TiO2 nanoparticles to the water and produce an increment in the concentration of H2O2 of 270 nM/day. Our results, together with the data provided by tourism records in the Mediterranean, point to TiO2 nanoparticles as the major oxidizing agent entering coastal waters, with direct ecological consequences on the ecosystem.


Enhanced immunogenicity of CD4(+) t-cell responses and protective efficacy of a DNA-modified vaccinia virus Ankara prime-boost vaccination regimen for murine tuberculosis.
DNA vaccines whose DNA encodes a variety of antigens from Mycobacterium tuberculosis have been evaluated for immunogenicity and protective efficacy. CD8(+) T-cell responses and protection achieved in other infectious disease models have been optimized by using a DNA immunization to prime the immune system and a recombinant virus encoding the same antigen(s) to boost the response. A DNA vaccine (D) and recombinant modified vaccinia virus Ankara (M) in which the DNA encodes early secreted antigenic target 6 and mycobacterial protein tuberculosis 63 synthesized, and each was found to generate specific gamma interferon (IFN-gamma)-secreting CD4(+) T cells. Enhanced CD4(+) IFN-gamma T-cell responses were produced by both D-M and M-D immunization regimens. Significantly higher levels of IFN-gamma were seen with a D-D-D-M immunization regimen. The most immunogenic regimens were assessed in a challenge study and found to produce protection equivalent to that produced by Mycobacterium bovis BCG. Thus, heterologous prime-boost regimens boost CD4(+) as well as CD8(+) T-cell responses, and the use of heterologous constructs encoding the same antigen(s) may improve the immunogenicity and protective efficacy of DNA vaccines against tuberculosis and other diseases.


How will South Africa's mandatory salt reduction policy affect its salt iodisation programme? A cross-sectional analysis from the WHO-SAGE Wave 2 Salt & Tobacco study.
The WHO's global targets for non-communicable disease reduction recommend consumption of<5 g salt/day. In 2016, South Africa was the first country to legislate maximum salt levels in processed foods. South Africa's salt iodisation fortification programme has successfully addressed iodine deficiency but information is dated. Simultaneous monitoring of sodium reduction and iodine status is required to ensure compatibility of the two public health interventions. A nested cohort design within WHO's 2015 Study on global AGEing and adult health (n=2887) including individuals from households across South Africa. Randomly selected adults (n=875) provided 24-hour and spot urine samples for sodium and iodine concentration analysis (the primary and secondary outcome measures, respectively). Median 24-hour urinary iodine excretion (UIE) and spot urinary iodine concentrations (UIC) were compared by salt intakes of <5g/day, 5-9g/dayand >9 g/day. Median daily sodium excretion was equivalent to 6.3 g salt/day (range 1-43 g/day); 35% had urinary sodium excretion values within the desirable range (<5 g salt/day), 37% had high values (5-9 g salt/day) and 28% had very high values (>9 g salt/day). Median UIC was 130 µg/L (IQR=58-202), indicating population iodine sufficiency (≥100 µg/L). Both UIC and UIE differed across salt intake categories (p<0.001) and were positively correlated with estimated salt intake (r=0.166 and 0.552, respectively; both p<0.001). Participants with salt intakes of <5 g/day were not meeting the Estimated Average Requirement for iodine intake (95 µg/day). In a nationally representative sample of South African adults, the association between indicators of population iodine status (UIC and UIE) and salt intake, estimated using 24-hour urinary sodium excretion, indicate that low salt intakes may compromise adequacy of iodine intakes in a country with mandatory iodisation of table salt. The iodine status of populations undergoing salt reduction strategies needs to be closely monitored to prevent re-emergence of iodine deficiency.


Chromosome 1 Sequence Analysis of C57BL/6J-Chr1KM Mouse Strain.
The Chinese Kunming (KM) mouse is a widely used outbred mouse stock in China. However, its genetic structure remains unclear. In this study, we sequenced the genome of the C57BL/6J-Chr1KM (B6-Chr1KM) strain, the chromosome 1 (Chr 1) of which was derived from one KM mouse. With 36.6× average coverage of the entire genome, 0.48 million single nucleotide polymorphisms (SNPs) and 96,679 indels were detected on Chr 1 through comparison with reference strain C57BL/6J. Moreover, 46,590 of them were classified as novel mutations. Further functional annotation identified 155 genes harboring potentially functional variants, among which 27 genes have been associated with human diseases. We then performed sequence similarity and Bayesian concordance analysis using the SNPs identified on Chr 1 and their counterparts in three subspecies, Mus musculus domesticus, M. m. musculus, and M. m. castaneus. Both analyses suggested that the Chr 1 sequence of B6-Chr1KM was predominantly derived from M. m. domesticus while 9.7% of the sequence was found to be from M. m. musculus. In conclusion, our analysis provided a detailed description of the genetic variations on Chr 1 of B6-Chr1KM and a new perspective on the subspecies origin of KM mouse which can be used to guide further genetic studies with this mouse strain.


Community attitudes to mental illness.
The baseline findings from a controlled study of the effect of a public education campaign on community attitudes to mental illness are presented. A census of attitudes to mental illness was conducted in two areas, prior to the opening of supported houses for the mentally ill. Factor analysis of the Community Attitudes toward the Mentally Ill (CAMI) inventory revealed three components: Fear and Exclusion, Social Control and Goodwill. The only determinant of Fear and Exclusion was having children. The main determinants of Social Control were social class, ethnic origin, age, having suffered mental illness and having children. The main determinant of Goodwill was educational level. The attitude factors were predictive of respondents' behavioural intentions toward the mentally ill. Respondents with children and non-Caucasians were more likely to object to the mentally ill living in their neighbourhood. Any intervention aimed at changing attitudes to mentally ill people in the community should be targeted at people with children and non-Caucasians, as these groups are more likely to object.


Transfection of wild-type TP53 induces differentiation in human gingival carcinoma cells.
We investigated the effects of transfection of wild-type TP53 on the growth properties of a human gingival carcinoma cell line, KOSC-3, in which the TP53 gene is mutated at codon 248 and overexpressed. The wild-type TP53 expression plasmid, pCDM8-p53/neo and the control plasmid, pCDM8/neo, were each stably transfected into KOSC-3 cells by using the calcium phosphate method. The number of G418-resistant colonies from wild-type TP53-transfected cells was approximately half that from plasmid controls. Exogenous wild-type TP53 transcripts were identified in four of the 20 G418-resistant clones analysed by reverse transcription PCR. Although the growth rates of the wild-type TP53+ clones did not drastically change during log phase, their saturation density was significantly reduced. The wild-type TP53+ cells were morphologically flat and enlarged when cultured in vitro, and were less able to form colonies in soft agar. In nude mice, the wild-type TP53+ clones formed subcutaneous tumours with conspicuous keratinisation and notable cell death that was not manifested in the parental and plasmid control cells. These findings indicate that the wild-type TP53 gene, even when it coexists with a mutated form, may function as a growth suppressor and differentiation inducer under restricted conditions in gingival squamous cell carcinoma.


Increased Expression of CD81 in Breast Cancer Tissue is Associated with Reduced Patient Prognosis and Increased Cell Migration and Proliferation in MDA-MB-231 and MDA-MB-435S Human Breast Cancer Cell Lines In Vitro.
BACKGROUND CD81, a member of the tetraspanin family, is overexpressed in several tumor types, but its role in breast cancer remains unknown. The present study aimed to investigate the effects of increased CD81 expression on cell migration and proliferation in breast cancer cell lines, MDA-MB-231 and MDA-MB-435S in vitro, and the effects of increased CD81 expression in breast cancer tissue microarrays on patient prognosis. MATERIAL AND METHODS The expression of CD81 was evaluated using immunohistochemistry in a human breast cancer tissue microarray containing 140 tumor tissues and a microarray containing 77 normal breast tissues. The effects of increased CD81 expression on cell proliferation and migration in breast cancer cell lines, MDA-MB-231 and MDA-MB-435S, were evaluated by proliferation, transwell migration, and cell counting kit-8 (CCK-8) assays. CD81-expressing plasmid transfection upregulated CD81 expression, and short hairpin RNA (shRNA) lentivirus silenced CD81 expression in vitro. RESULTS CD81 expression was significantly increased in breast cancer tissues compared with normal breast tissues (P<0.05). Increased expression of CD81 was significantly associated with lymph node metastasis (P<0.05), clinical stage (P<0.05) and with reduced overall survival (OS) in patients with breast cancer (P<0.05). Increased CD81 expression in MDA-MB-231 and MDA-MB-435S cells promoted cell proliferation and migration, which were inhibited by CD81 silencing. CONCLUSIONS The findings of the study showed that CD81 might be a potential prognostic biomarker associated with poor patient prognosis in breast cancer. These findings should be investigated further with large-scale controlled prospective studies in patients with breast cancer.


The effect of restricting the indications for lumbosacral spine radiography in patients with acute back symptoms.
A prospective study was performed to evaluate the effect of using a special requisition form on the utilization of lumbosacral spine radiography for patients presenting in the emergency room with acute lower back complaints. Over a 1-year period, emergency room house officers were encouraged to complete a special form that listed only three acceptable indications for lumbosacral spine radiographs: history of trauma, evidence of focal neurologic abnormality, and "other." Neurologic abnormalities included hypesthesia, hyperesthesia or anesthesia of lumbar or sacral dermatomes, weakness or hyporeflexia of the lower extremities, and bladder or bowel incontinence. If the indication "other" was chosen, radiographs were done only if the form contained both a one- to two-sentence history and the signed approval of a supervising attending physician. The number and results of lumbosacral spine examinations were compared with those from the previous year, which served as a control. In the control year 1443 examinations were performed, and in the experimental year only 759 were done (a decrease of 47%). The percentage of patients with vertebral fractures increased from 5.1% to 5.8% in the experimental year if only the detection of new fractures was considered positive, and from 9.1% to 13.4% if the detection of fractures of all ages was regarded as significant. The use of the special requisition form appears to be a simple and effective means of reducing unnecessary lumbosacral radiography in the emergency room setting.


Reperfusion-induced arrhythmias. A role for washout of extracellular protons?
Rapid washout of extracellular H+ accumulated during preceding ischemia (i.e., the abrupt restoration of extracellular pH) has been implicated as an arrhythmogenic factor during reperfusion. Therefore, we hypothesized that by limiting the rate at which extracellular pH was restored during early reperfusion it should be possible to protect against reperfusion-induced arrhythmias. To test this, we used isolated rat hearts (n = 12 per group) and a novel dual coronary perfusion cannula that permitted the induction of regional ischemia (10 minutes) and the selective reperfusion (8 minutes) of the ischemic zone with modified solutions. We examined the antiarrhythmic efficacy of 1) acidic (pH 6.6) reperfusion with stepwise restoration of extracellular pH to 7.4 (stepped pH) and 2) transient (2-minute) acidic (pH 7.1, 6.8, 6.6, or 6.4) reperfusion with subsequent abrupt restoration of extracellular pH to 7.4. Hearts in two contemporary control groups were reperfused with solution at pH 7.4 throughout. In all groups, 100% of hearts exhibited ventricular tachycardia (VT) on reperfusion. VT degenerated into ventricular fibrillation (VF) in 100% of hearts in the control group but in only 42% of hearts in the stepped-pH group (p < 0.05). In the groups subjected to transient acidic reperfusion, there was a pH-dependent prolongation of VT cycle length (measured at 15 seconds of reperfusion), which was 47.1 +/- 3.9, 51.1 +/- 5.5, 56.0 +/- 1.9, 60.4 +/- 2.8 (p < 0.05), and 68.8 +/- 5.0 (p < 0.05) msec in the pH 7.4 (control), 7.1, 6.8, 6.6, and 6.4 groups, respectively. In these groups, VT degenerated into VF in 92%, 92%, 83%, 42% (p < 0.05), and 33% (p < 0.05) of hearts, respectively. In conclusion, limiting the rate at which extracellular pH is restored during early reperfusion does not affect the rapid induction of VT but inhibits the degeneration of VT into VF and promotes spontaneous reversion to normal sinus rhythm. This is consistent with a major arrhythmogenic role, during uncontrolled reperfusion, for the rapid washout of extracellular H+.


Understanding the regulation of estivation in a freshwater snail through iTRAQ-based comparative proteomics.
The apple snail Pomacea canaliculata is a freshwater gastropod with a remarkable ability to withstand seasonal or unpredictable dry conditions by entering estivation. Studies of P. canaliculata using conventional biochemical and the individual gene approaches have revealed the expressional changes of several enzymes and antioxidative genes in response to estivation and arousal. In this study, we applied iTRAQ-coupled two-dimensional LC-MS/MS to identify and quantify the global protein expression during the estivation and arousal of P. canaliculata. A total of 1040 proteins were identified, among which 701 proteins were quantified and compared across four treatments (i.e., control, active snails; short-term estivation, 3 days of exposure to air; prolonged estivation, 30 days of exposure to air; and arousal, 6 h after resubmergence in water) revealing 53 differentially expressed proteins. A comparison of protein expression profiles across treatments indicated that the proteome of this species was very insensitive to initial estivation, with only 9 proteins differentially expressed as compared with the control. Among the 9 proteins, the up-regulations of two immune related proteins indicated the initial immune response to the detection of stress cues. Prolonged estivation resulted in many more differentially expressed proteins (47 compared with short-term estivation treatment), among which 16 were down-regulated and 31 were up-regulated. These differentially expressed proteins have provided the first global picture of a shift in energy usage from glucose to lipid, prevention of protein degradation and elevation of oxidative defense, and production of purine for uric acid production to remove toxic ammonia during prolonged estivation in a freshwater snail. From prolonged estivation to arousal, only 6 proteins changed their expression level, indicating that access to water and food alone is not a necessary condition to reactivate whole-sale protein expression. A comparison with hibernation and diapause revealed many similar molecular mechanisms of hypometabolic regulation across the animal kingdom.


Microtubule minus-end stability is dictated by the tubulin off-rate.
Dynamic organization of microtubule minus ends is vital for the formation and maintenance of acentrosomal microtubule arrays. In vitro, both microtubule ends switch between phases of assembly and disassembly, a behavior called dynamic instability. Although minus ends grow slower, their lifetimes are similar to those of plus ends. The mechanisms underlying these distinct dynamics remain unknown. Here, we use an in vitro reconstitution approach to investigate minus-end dynamics. We find that minus-end lifetimes are not defined by the mean size of the protective GTP-tubulin cap. Rather, we conclude that the distinct tubulin off-rate is the primary determinant of the difference between plus- and minus-end dynamics. Further, our results show that the minus-end-directed kinesin-14 HSET/KIFC1 suppresses tubulin off-rate to specifically suppress minus-end catastrophe. HSET maintains its protective minus-end activity even when challenged by a known microtubule depolymerase, kinesin-13 MCAK. Our results provide novel insight into the mechanisms of minus-end dynamics, essential for our understanding of microtubule minus-end regulation in cells.


Loss-of-function mutations in Lysyl-tRNA synthetase cause various leukoencephalopathy phenotypes.
To expand the clinical spectrum of lysyl-tRNA synthetase (KARS) gene-related diseases, which so far includes Charcot-Marie-Tooth disease, congenital visual impairment and microcephaly, and nonsyndromic hearing impairment. Whole-exome sequencing was performed on index patients from 4 unrelated families with leukoencephalopathy. Candidate pathogenic variants and their cosegregation were confirmed by Sanger sequencing. Effects of mutations on KARS protein function were examined by aminoacylation assays and yeast complementation assays. Common clinical features of the patients in this study included impaired cognitive ability, seizure, hypotonia, ataxia, and abnormal brain imaging, suggesting that the CNS involvement is the main clinical presentation. Six previously unreported and 1 known KARS mutations were identified and cosegregated in these families. Two patients are compound heterozygous for missense mutations, 1 patient is homozygous for a missense mutation, and 1 patient harbored an insertion mutation and a missense mutation. Functional and structural analyses revealed that these mutations impair aminoacylation activity of lysyl-tRNA synthetase, indicating that defective KARS function is responsible for the phenotypes in these individuals. Our results demonstrate that patients with loss-of-function KARS mutations can manifest CNS disorders, thus broadening the phenotypic spectrum associated with KARS-related disease.


Fully compensated Michelson interferometer of fixed-path difference.
A Michelson interferometer of fixed-path difference, capable of scanning over only one fringe, has been shown to be useful for the measurement of Doppler shifts from isolated emission lines. It is straightforward to field widen this type of configuration, but if different wavelengths are to be selected, the field widening must be achromatic. Further, for Doppler shift measurement, one requires a path difference that is highly stable with respect to temperature. The generalized requirements for meeting all these conditions are set out and a configuration that satisfies them is described. Sample characteristics are given for a number of different systems.


Elevated Norepinephrine Stimulates Adipocyte Hyperplasia in Ovine Fetuses With Placental Insufficiency and IUGR.
Prevailing hypoxemia and hypoglycemia in near-term fetuses with placental insufficiency-induced intrauterine growth restriction (IUGR) chronically increases norepinephrine concentrations, which lower adrenergic sensitivity and lipid mobilization postnatally, indicating a predisposition for adiposity. To determine adrenergic-induced responses, we examined the perirenal adipose tissue transcriptome from IUGR fetuses with or without hypercatecholaminemia. IUGR was induced in sheep with maternal hyperthermia, and hypercatecholaminemia in IUGR was prevented with bilateral adrenal demedullation. Adipose tissue was collected from sham-operated control (CON) and IUGR fetuses and adrenal-demedullated control (CAD) and IUGR (IAD) fetuses. Norepinephrine concentrations were lower in IAD fetuses than in IUGR fetuses despite both being hypoxemic and hypoglycemic. In IUGR fetuses, perirenal adipose tissue mass relative to body mass was greater compared with the CON, adrenal-demedullated control, and IAD groups. Transcriptomic analysis identified 581 differentially expressed genes (DEGs) in CON vs IUGR adipose tissue and 193 DEGs in IUGR vs IAD adipose tissue. Integrated functional analysis of these 2 comparisons showed enrichment for proliferator-activated receptor signaling and metabolic pathways and identified adrenergic responsive genes. Within the adrenergic-regulated DEGs, we identified transcripts that regulate adipocyte proliferation and differentiation: adipogenesis regulatory factor, C/CCAAT/enhancer binding protein α, and sterol carrier protein 2. DEGs associated with the metabolic pathway included pyruvate dehydrogenase kinase 4, 6-phosphofructo-2-kinase/fructose-2,6-biphosphatase 4, IGF-binding proteins (IGFBP-5 and IGFBP-7). Sex-specific expression differences were also found for adipogenesis regulatory factor, pyruvate dehydrogenase kinase 4, IGFBP5, and IGFBP7. These findings indicate that sustained adrenergic stimulation during IUGR leads to adipocyte hyperplasia with alterations in metabolism, proliferation, and preadipocyte differentiation pathways.


Scoring Systems for Outcome Prediction in a Cardiac Surgical Intensive Care Unit: A Comparative Study.
Most scoring systems used to predict clinical outcome in critical care were not designed for application in cardiac surgery patients. To compare the predictive ability of the most widely used scoring systems (Acute Physiology and Chronic Health Evaluation [APACHE] II, Simplified Acute Physiology Score [SAPS] II, and Sequential Organ Failure Assessment [SOFA]) and of 2 specialized systems (European System for Cardiac Operative Risk Evaluation [EuroSCORE] II and the cardiac surgery score [CASUS]) for clinical outcome in patients after cardiac surgery. Consecutive patients admitted to a cardiac surgical intensive care unit (CSICU) were prospectively studied. Data on the preoperative condition, intraoperative parameters, and postoperative course were collected. EuroSCORE II, CASUS, and scores from 3 general severity-scoring systems (APACHE II, SAPS II, and SOFA) were calculated on the first postoperative day. Clinical outcome was defined as 30-day mortality and in-hospital morbidity. A total of 150 patients were included. Thirty-day mortality was 6%. CASUS was superior in outcome prediction, both in relation to discrimination (area under curve, 0.89) and calibration (Brier score = 0.043, χ(2) = 2.2, P = .89), followed by EuroSCORE II for 30-day mortality (area under curve, 0.87) and SOFA for morbidity (Spearman ρ= 0.37 and 0.35 for the CSICU length of stay and duration of mechanical ventilation, respectively; Wilcoxon W = 367.5, P = .03 for probability of readmission to CSICU). CASUS can be recommended as the most reliable and beneficial option for benchmarking and risk stratification in cardiac surgery patients.


Similarity of fibroglandular breast tissue content measured from magnetic resonance and mammographic images and by a mathematical algorithm.
Women with high breast density (BD) have a 4- to 6-fold greater risk for breast cancer than women with low BD. We found that BD can be easily computed from a mathematical algorithm using routine mammographic imaging data or by a curve-fitting algorithm using fat and nonfat suppression magnetic resonance imaging (MRI) data. These BD measures in a strictly defined group of premenopausal women providing both mammographic and breast MRI images were predicted as well by the same set of strong predictor variables as were measures from a published laborious histogram segmentation method and a full field digital mammographic unit in multivariate regression models. We also found that the number of completed pregnancies, C-reactive protein, aspartate aminotransferase, and progesterone were more strongly associated with amounts of glandular tissue than adipose tissue, while fat body mass, alanine aminotransferase, and insulin like growth factor-II appear to be more associated with the amount of breast adipose tissue. Our results show that methods of breast imaging and modalities for estimating the amount of glandular tissue have no effects on the strength of these predictors of BD. Thus, the more convenient mathematical algorithm and the safer MRI protocols may facilitate prospective measurements of BD.


Can biological activity be maintained at ultra-high dilution? An overview of homeopathy, evidence, and Bayesian philosophy.
Forty percent of general practitioners in the Netherlands practice homeopathy. With over 100 homeopathic medical schools, homeopathy is practiced in India along with conventional Western medicine in government clinics. In Britain, 42% of general practitioners refer patients to homeopaths. Two recent meta-analyses of homeopathy both indicate that there is enough evidence to show that homeopathy has added effects over placebo. Against this evidence is a backdrop of considerable scientific scepticism. Homeopathic remedies are diluted substances--some are so diluted that statistically there are no molecules present to explain their proposed biological effects (ultra-high dilutions or UHDs). Without knowledge of the evidence, most scientists would reject UHD effects because of their intrinsic implausibility in the light of our current scientific understanding. The objective of this article is to critically review the major pieces of evidence on UHD effects and suggest how the scientific community should respond to its challenge. Such evidence has been conducted on a diverse range of assays--immunologic, physiological, behavioral, biochemical, and clinical in the form of trials of homeopathic remedies. Evidence of UHD effects has attracted the attention of physicists who have speculated on their physical mechanisms. Included is a critique of several experiments that form the Benveniste affair which was sparked by a publication in Nature that advocated the existence of UHD effects of anti-immunoglobulin E (IgE) on human basophils, and is the paradigm example of how a controversial phenomenon can split the scientific community. It is argued that if the phenomenon was uncontroversial, the evidence suffices to show that UHD effects exist. However, given that the observations contradict well-established theory, normal science has to be abandoned and scientists need to decide for themselves what the likelihood of UHD effects are. Bayesian analysis describes how scientists ought rationally to change their prior beliefs in the light of evidence. Theories by Kuhn and Lakatos indicate that whether UHD effects are proved or not depends on the beliefs and behaviors of scientists in their communities. This article argues that there is as yet insufficient evidence to drive rational scientists to a consensus over UHD effects, even if they possessed knowledge of all the evidence. The difficulty in publishing high-quality UHD research in conventional journals prevents a fair assessment of UHD effects. Given that the existence of UHD effects would revolutionize science and medicine, and given the considerable empirical evidence of them, the philosophies of science tell us that possible UHD effects warrant serious investigation by conventional scientists and serious attention by scientific journals.


A human origin strain Lactobacillus acidophilus DDS-1 exhibits superior in vitro probiotic efficacy in comparison to plant or dairy origin probiotics.
Background: The health benefits of probiotics are well established and known to be strain-specific. However, the role of probiotics obtained from different origins and their efficacy largely remains unexplored. The aim of this study is to investigate the in vitro efficacy of probiotics from different origins. Methods: Probiotic strains utilized in this study include Lactobacillus acidophilus DDS-1 (human origin), Bifidobacterium animalis ssp. lactis UABla-12 (human origin), L. plantarum UALp-05 (plant origin) and Streptococcus thermophilus UASt-09 (dairy origin). Screening assays such as in vitro digestion simulation, adhesion, cell viability and cytokine release were used to evaluate the probiotic potential. Results: All strains showed good resistance in the digestion simulation process, especially DDS-1 and UALp-05, which survived up to a range of 107 to 108 CFU/mL from an initial concentration of 109 CFU/mL. Two human colonic mucus-secreting cells, HT-29 and LS174T, were used to assess the adhesion capacity, cytotoxicity/viability, and cytokine quantification. All strains exhibited good adhesion capacity. No significant cellular cytotoxicity or loss in cell viability was observed. DDS-1 and UALp-05 significantly upregulated anti-inflammatory IL-10 and downregulated pro-inflammatory TNF-α cytokine production. All the strains were able to downregulate IL-8 cytokine levels. Conclusion: Of the 4 strains tested, DDS-1 demonstrated superior survival rates, good adhesion capacity and strong immunomodulatory effect under different experimental conditions.


Systemic analysis of heat shock response induced by heat shock and a proteasome inhibitor MG132.
The molecular basis of heat shock response (HSR), a cellular defense mechanism against various stresses, is not well understood. In this, the first comprehensive analysis of gene expression changes in response to heat shock and MG132 (a proteasome inhibitor), both of which are known to induce heat shock proteins (Hsps), we compared the responses of normal mouse fibrosarcoma cell line, RIF-1, and its thermotolerant variant cell line, TR-RIF-1 (TR), to the two stresses. The cellular responses we examined included Hsp expressions, cell viability, total protein synthesis patterns, and accumulation of poly-ubiquitinated proteins. We also compared the mRNA expression profiles and kinetics, in the two cell lines exposed to the two stresses, using microarray analysis. In contrast to RIF-1 cells, TR cells resist heat shock caused changes in cell viability and whole-cell protein synthesis. The patterns of total cellular protein synthesis and accumulation of poly-ubiquitinated proteins in the two cell lines were distinct, depending on the stress and the cell line. Microarray analysis revealed that the gene expression pattern of TR cells was faster and more transient than that of RIF-1 cells, in response to heat shock, while both RIF-1 and TR cells showed similar kinetics of mRNA expression in response to MG132. We also found that 2,208 genes were up-regulated more than 2 fold and could sort them into three groups: 1) genes regulated by both heat shock and MG132, (e.g. chaperones); 2) those regulated only by heat shock (e.g. DNA binding proteins including histones); and 3) those regulated only by MG132 (e.g. innate immunity and defense related molecules). This study shows that heat shock and MG132 share some aspects of HSR signaling pathway, at the same time, inducing distinct stress response signaling pathways, triggered by distinct abnormal proteins.


Vasoactive side effects of intravenous immunoglobulin preparations in a rat model and their treatment with recombinant platelet-activating factor acetylhydrolase.
Previously, we observed in a rat model that intravenous administration of intramuscular immunoglobulin preparations induced a long-lasting hypotension, which appeared to be associated with the presence of IgG polymers and dimers in the preparations, but unrelated to complement activation. We found evidence that this hypotensive response is mediated by platelet-activating factor (PAF) produced by macrophages. In this study, we compared the vasoactive effects of 16 intravenous immunoglobulin (IVIG) products from 10 different manufacturers, in anesthetized rats. Eight of the IVIG preparations showed no hypotensive effects (less than 15% decrease), whereas the other 8 had relatively strong effects (15%-50% decrease). The hypotensive effects correlated with the IgG dimer content of the preparations. Pretreatment of the rats with recombinant PAF acetylhydrolase completely prevented the hypotensive reaction on IVIG infusion, and administration after the onset of hypotension resulted in normalization of the blood pressure. We also observed PAF production on in vitro incubation of human neutrophils with IVIG, which could be blocked by anti-Fcgamma receptor antibodies. This indicates that induction of PAF generation may also occur in a human system. Our findings support the hypothesis that the clinical side effects of IVIG in patients may be caused by macrophage and neutrophil activation through interaction of IgG dimers with Fcgamma receptors. Because phagocyte activation may also lead to the release of other inflammatory mediators, recombinant PAF acetylhydrolase (rPAF-AH) provides a useful tool to determine whether PAF plays a role in the clinical side effects of IVIG. If so, rPAF-AH can be used for the treatment of those adverse reactions. (Blood. 2000;95:1856-1861)


Adaptation to the deleterious effects of antimicrobial drug resistance mutations by compensatory evolution.
Compensatory mutations, due to their ability to mask the deleterious effects of another mutation, are important for the adaptation and evolution of most organisms. Resistance to antibiotics, antivirals, antifungals, herbicides and insecticides is usually associated with a fitness cost. As a result of compensatory evolution, the initial fitness costs conferred by resistance mutations (or other deleterious mutations) can often be rapidly and efficiently reduced. Such compensatory evolution is potentially of importance for (i) the long-term persistence of drug resistance, (ii) reducing the rate of fitness loss associated with the accumulation of deleterious mutations in small asexual populations, and (iii) the evolution of complexity of cellular processes.


The 5'-terminal region of a tombusvirus genome determines the origin of multivesicular bodies.
Multivesicular bodies (MVB) are membranous cytoplasmic inclusions that are invariably associated with tombusvirus infections regardless of the virus species, the host, or the tissue examined. MVB are virus-induced structures since they are absent from tissues of healthy plants and are always present both in infected plants and protoplasts. MVB derive from peroxisomes in cells infected by a number of tombusviruses including cymbidium ringspot virus (CymRSV) and from mitochondria in cells infected by another tombusvirus, carnation Italian ringspot virus (CIRV). By using common restriction sites in full-length infectious clones, hybrid clones of these two viruses were constructed. In addition, a mutant of CIRV was prepared in which the protein encoded by the first open reading frame was shortened by 22 amino acids. All mutant transcripts were viable and infected Nicotiana benthamiana plants. Infected leaf tissue samples were collected, processed for thin sectioning, and observed in the electron microscope. The origin of MVB was shown to be under the control of the 5' region of the viral genome. A sequence as short as about 600 nucleotides in ORF 1 contained the determinants for formation of MVB from peroxisomes or mitochondria.


Endocannabinoid identification in the brain: studies of breakdown lead to breakthrough, and there may be NO hope.
Endocannabinoids are a class of fatty acid derivatives defined by their ability to interact with the specific cannabinoid receptors that were originally identified as the targets of Delta9-tetrahydocannabinol (Delta9-THC), the psychoactive component of cannabis. Endocannabinoids have been implicated in a growing number of important physiological and behavioral events. A full understanding of the functions of endocannabinoids will involve knowing which ones are active, and how they are produced, during any given physical event. However, studying these small lipids in the brain presents many technical challenges. New selective pharmacological tools promise to be very useful in unraveling the complexities of endocannabinoid signaling, but parallel developments from the investigation of the cellular neurophysiology of the endocannabinoid systems highlight the difficulties remaining.


New Daily Persistent Headache and OnabotulinumtoxinA Therapy.
The aim of the study was to determine whether onabotulinumtoxinA therapy is effective in the treatment of new daily persistent headache (NDPH). New daily persistent headache is a difficult to treat headache syndrome resistant to both conventional and unconventional headache therapies. New daily persistent headache was excluded in the registration trials for onabotunlinumtoxinA (onabot) in chronic migraine. Apart from case reports supporting its benefit, little is known about its therapeutic value in NDPH. We performed a single-center, retrospective chart review of patients with a diagnosis of NDPH who received onabot treatment for a 30-month period at the Cleveland Clinic Headache Center. Measures of interest were headache frequency and headache severity. All patients had received the Food and Drug Administration-approved PREEMPT Protocol. A decrease in headache frequency was noted in 8 (50.0%) of 16 patients at 6 months and 7 (63.6%) of 11 patients at 12 months. Headache severity improved in 5 (50.0%) of 10 patients at 6 months and in 7 (77.8%) of 9 patients at 12 months. Most therapies are unable to break the unremitting course of NDPH. In our investigation, at 1 year (3-4 cycles of onabot treatment), approximately half of the patients treated showed a reduction in headache frequency and approximately 75% demonstrated some improvement in headache severity. Evidence from this small-scale retrospective study suggests that onabot shows strong promise for the treatment of NDPH, which currently is resistant to most therapies, but a randomized controlled study should be the next step in confirmation of this therapy.


Deficiency of the ADP-forming succinyl-CoA synthase activity is associated with encephalomyopathy and mitochondrial DNA depletion.
The mitochondrial DNA (mtDNA) depletion syndrome is a quantitative defect of mtDNA resulting from dysfunction of one of several nuclear-encoded factors responsible for maintenance of mitochondrial deoxyribonucleoside triphosphate (dNTP) pools or replication of mtDNA. Markedly decreased succinyl-CoA synthetase activity due to a deleterious mutation in SUCLA2, the gene encoding the beta subunit of the ADP-forming succinyl-CoA synthetase ligase, was found in muscle mitochondria of patients with encephalomyopathy and mtDNA depletion. Succinyl-CoA synthetase is invariably in a complex with mitochondrial nucleotide diphosphate kinase; hence, we propose that a defect in the last step of mitochondrial dNTP salvage is a novel cause of the mtDNA depletion syndrome.


Independent prognostic factors for AML outcome.
Over the last three decades there have been dramatic advances in deciphering the cytogenetic and molecular lesions underlying the pathogenesis of acute myeloid leukemia (AML). These have not only afforded greater insights into disease biology, but also provided useful information predicting the likelihood of any given patient achieving and maintaining remission following conventional chemotherapy, leading to the development of risk-stratified treatment approaches. However, it is becoming increasingly apparent that AML is highly heterogeneous at the molecular level. Defining the individual genetic abnormalities or combinations of markers that provide significant independent prognostic information and establishing their respective relationships to other pre-treatment characteristics that impact on outcome, such as age and presenting white blood cell count, presents a major ongoing challenge. Moreover, there is increasing evidence that risk of relapse and overall survival can be predicted by assessment of kinetics and depth of response following front-line therapy and monitoring of the leukemic burden using molecular or immunological approaches to minimal residual disease (MRD) detection. These advances present the exciting prospect that panels of pre-treatment parameters affording independent prognostic information can be integrated with precise measurement of treatment response using MRD technologies to provide greater refinement in risk-adapted management of AML. This could lead to further improvements in outcome and serve to identify in a more reliable fashion those patients most likely to benefit from allogeneic transplant in first remission.


The PGE2/IL-10 Axis Determines Susceptibility of B-1 Cell-Derived Phagocytes (B-1CDP) to Leishmania major Infection.
B-1 cells can be differentiated from B-2 cells because they are predominantly located in the peritoneal and pleural cavities and have distinct phenotypic patterns and activation properties. A mononuclear phagocyte derived from B-1 cells (B-1CDP) has been described. As the B-1CDP cells migrate to inflammatory/infectious sites and exhibit phagocytic capacity, the microbicidal ability of these cells was investigated using the Leishmania major infection model in vitro. The data obtained in this study demonstrate that B-1CDP cells are more susceptible to infection than peritoneal macrophages, since B-1CDP cells have a higher number of intracellular amastigotes forms and consequently release a larger number of promastigotes. Exacerbated infection by L. major required lipid bodies/PGE2 and IL-10 by B-1CDP cells. Both infection and the production of IL-10 were decreased when PGE2 production was blocked by NSAIDs. The involvement of IL-10 in this mechanism was confirmed, since B-1CDP cells from IL-10 KO mice are more competent to control L. major infection than cells from wild type mice. These findings further characterize the B-1CDP cells as an important mononuclear phagocyte that plays a previously unrecognized role in host responses to L. major infection, most likely via PGE2-driven production of IL-10.


Risk of spontaneous preterm birth is associated with common proinflammatory cytokine polymorphisms.
Preliminary data suggest that common genetic variation in immune response genes can contribute to the risk for spontaneous preterm birth and possibly small-for-gestational age (SGA). We investigated the relationship of polymorphisms in 6 cytokine genes associated with inflammation-interleukin (IL)1alpha, IL1beta, IL2, IL6, tumor necrosis factor (TNF), and lymphotoxin alpha (LTA)-with spontaneous preterm and SGA birth in a nested case-control study drawn from a prospective pregnancy cohort. Women were recruited between 24 and 29 weeks' gestation at the Wake County and University of North Carolina, Chapel Hill obstetric clinics between February 1996 and June 2000. We inferred haplotypes using the EM algorithm and the Bayesian method, PHASE. We then compared haplotype frequency distributions and implemented semi-Bayesian hierarchical logistic regression analyses to obtain odds ratio (OR) estimates and 95% confidence intervals (CIs) for each polymorphism. Two haplotypes spanning the TNF/LTA genes were associated with increased risk for spontaneous preterm birth in white subjects (for the AGG haplotype, OR = 1.5 [95% CI=0.8-2.6]; for the GAC haplotype, 1.6 [0.9-2.9]). Additionally, carriers of the GAG haplotype were found to have decreased risk of spontaneous preterm birth (0.6; 0.3-1.0). The TNF(-488)A and LTA(IVS1-82)C variants, constituents of the AGG and GAC haplotypes respectively, were also strongly associated with increased risk of spontaneous preterm birth. Our results suggest that common genetic variants in proinflammatory cytokine genes could influence the risk for spontaneous preterm birth. Selected TNF/LTA haplotypes were associated with spontaneous preterm birth in both African-American and white subjects. Our data do not support an inflammatory etiology for SGA.


Approaches to advancing blood safety through haemovigilance: a review.
Blood transfusion is always associated with some level of risk. Haemovigilance is a risk monitoring system integral to the practice of transfusion medicine whose ultimate purpose is to improve the quality and safety of transfusion therapy. To examine the contribution of haemovigilance to blood safety, including the approaches that some countries have taken to institute haemovigilance, and explore routes through which countries without such systems can achieve them. The internet and journals on the topic of haemovigilance and development of haemovigilance systems in the English language. Reputable journals on the topic of haemovigilance were examined for abstracts and papers. Abstracts based on known credible and distinguished sources were selected. Information on haemovigilance and the processes of developing haemovigilance in various countries was reviewed. The information from selected papers and abstracts was used for writing this paper. Varying processes for haemovigilance have been adopted by different countries. The more advanced systems have national/regional coordinating mechanisms. Availability of haemovigilance data has given transfusion services a clear understanding of problems associated with transfusion that need to be solved so as to improve transfusion safety. Although countries in sub-Saharan Africa have made considerable progress in enhancing blood safety in the recent past, nationally coordinated haemovigilance systems are lacking. Focus on haemovigilance systems is considered the next frontier to be conquered in enhancing blood safety in the region.


Potential impact of a 2-person security rule on BioSafety Level 4 laboratory workers.
Directors of all major BioSafety Level 4 (BSL-4) laboratories in the United States met in 2008 to review the current status of biocontainment laboratory operations and to discuss the potential impact of a proposed 2-person security rule on maximum-containment laboratory operations. Special attention was paid to the value and risks that would result from a requirement that 2 persons be physically present in the laboratory at all times. A consensus emerged indicating that a video monitoring system represents a more efficient, economical standard; provides greater assurance that pathogens are properly manipulated; and offers an increased margin of employee safety and institutional security. The 2-person security rule (1 to work and 1 to observe) may decrease compliance with dual responsibilities of safety and security by placing undue pressure on the person being observed to quickly finish the work, and by placing the observer in the containment environment unnecessarily.


The impact of the European financial crisis on clinical research within the European union or "when life gives you lemons, make lemonade".
The European Union (EU) and the world are faced with unprecedented economic challenges, which if allowed to persist could threaten its continued existence in its current form as a union. Furthermore, these same financial challenges can easily translate into societal hardship. The biomedical and pharmaceutical industries of the U.S. and Europe have been in the midst of severe financial constraints over the last few years, which will worsen in the coming years. A critical factor is a financial crisis that appears to be spreading like wildfire through Europe, with 3 of its oldest members (Greece, Ireland, Portugal) already having enlisted the aid of the International Monetary Fund (IMF) to avoid bankruptcy, and with the possibility of others to follow (Italy, Spain). Adding to this the increasing cost of introducing new medications and devices, and conducting clinical research, as well as the tightening regulatory pressures and the drying pharmaceutical pipelines, cost cutting pressures are mounting on pharmaceutical industries and the biomedical sector, whether in academia or in industry. This paper will attempt to highlight some of the problems that clinical research in Europe may be facing, as well as discuss some of the potential solutions. Although this will by no means be an exhaustive analysis, the goal is to show that times of financial hardship, such as the current one, may also provide the impetus for constructive change.


Evolution of an influenza pandemic in 13 countries from 5 continents monitored by protein microarray from neonatal screening bloodspots.
Because of lack of worldwide standardization of influenza virus surveillance, comparison between countries of impact of a pandemic is challenging. For that, other approaches to allow internationally comparative serosurveys are welcome. Here we explore the use of neonatal screening dried blood spots to monitor the trends of the 2009 influenza A (H1N1) pdm virus by the use of a protein microarray. We contacted colleagues from neonatal screening laboratories and asked for their willingness to participate in a study by testing anonymized neonatal screening bloodspots collected during the course of the pandemic. In total, 7749 dried blood spots from 13 countries in 5 continents where analyzed by using a protein microarray containing HA1 recombinant proteins derived from pandemic influenza A (H1N1) 2009 as well as seasonal influenza viruses. Results confirm the early start of the pandemic with extensive circulation in the US and Canada, when circulation of the new virus was limited in other parts of the world. The data collected from sites in Mexico suggested limited circulation of the virus during the early pandemic phase in this country. In contrast and to our surprise, an increase in seroprevalence early in 2009 was noted in the dataset from Argentina, suggestive of much more widespread circulation of the novel virus in this country than in Mexico. We conclude that this uniform serological testing of samples from a highly standardized screening system offers an interesting opportunity for monitoring population level attack rates of widespread diseases outbreaks and pandemics.


Experiences of Mothers on Parenting Children with Leukemia.
Childhood cancer is the leading cause of death among children. Leukemia is one of the most common childhood cancers. The objective of this study was to explore the experiences of mothers on parenting children with leukemia. A qualitative approach with phenomenological design was used. To collect depth information from the mothers of children with leukemia, purposive sampling technique was adopted. Data were collected from ten mothers. Semi-structured interview schedule was used to collect the data. Data were analyzed using Husserl's method. The themes derived are the pivotal moment in life, the experience of being with a seriously ill child, having to keep distance with the relatives, overcoming the financial and social commitments, responding to challenges, experience of faith as being key to survival, health concerns of the present and future, and optimism. The study concluded that chronic illness such as leukemia in children results in negative impact on the child and on the mother.


Ixr1, a yeast protein that binds to platinated DNA and confers sensitivity to cisplatin.
Structure-specific recognition proteins (SSRPs) bind to DNA containing intrastrand cross-links formed by the anticancer drug cisplatin. A yeast gene encoding an SSRP, designated IXR1, was cloned and sequenced. The Ixr1 protein, a member of the high mobility group-box protein family, bound specifically to DNA modified with cisplatin but not inactive platinum compounds. A yeast strain with an inactivated IXR1 gene was half as sensitive to cisplatin and accumulated one-third as many platinum-DNA lesions after treatment with cisplatin as the parental strain. These findings suggest that SSRPs play a role in mediating the cytotoxicity of cisplatin.


Efficacy of structural-level condom distribution interventions: a meta-analysis of U.S. and international studies, 1998-2007.
This systematic review examines the overall efficacy of U.S. and international-based structural-level condom distribution interventions (SLCDIs) on HIV risk behaviors and STIs and identifies factors associated with intervention efficacy. A comprehensive literature search of studies published from January 1988 through September 2007 yielded 21 relevant studies. Significant intervention effects were found for the following outcomes: condom use, condom acquisition/condom carrying, delayed sexual initiation among youth, and reduced incident STIs. The stratified analyses for condom use indicated that interventions were efficacious for various groups (e.g., youth, adults, males, commercial sex workers, clinic populations, and populations in areas with high STI incidence). Interventions increasing the availability of or accessibility to condoms or including additional individual, small-group or community-level components along with condom distribution were shown to be efficacious in increasing condom use behaviors. This review suggests that SLCDIs provide an efficacious means of HIV/STI prevention.


First Report of Ilyonectria macrodidyma Causing Root Rot of Olive Trees (Olea europaea) in California.
The California olive industry produces 99% of the U.S. olive crop, which represented a value of over $113 million in 2010. During the 2008 and 2009 growing seasons, decline of young super-high-density olive cvs. Arbequina, Arbosana, and Koroneiki trees (<4 years old) was observed in orchards throughout Glenn, Yolo, and San Joaquin Counties. Symptomatic trees showed stunted growth and chlorotic leaves with roots having black, sunken, necrotic lesions, which frequently prolonged into the base and crown of the tree. Twenty-five trees were collected from different orchards and necrotic roots as well as infected trunk tissue were plated onto potato dextrose agar amended with 0.01% tetracycline hydrochloride. Cultures were incubated at room temperature (23 ± 2°C) until fungal colonies were observed. In 17 out of 25 trees collected (68%), light yellow fungal colonies were observed from the symptomatic tissue after 7 to 10 days. Colonies turned dark yellow to orange with age and showed an orange-dark brown reverse. Both microconidia (hyaline, ellipsoidal to ovoidal and aseptate (n = 60) (6.5) 11.5 to 13.5 (17.1) × (3) 3.4 to 4.5 (5.6) μm) and macroconidia (hyaline, cylindrical, straight and/or slightly curved with one, two or three septa (n = 60) (12.5) 26.5 to 38.5 (44.1) × (4) 5.5 to 7.5 (8.5) μm) were observed. Culture and conidial morphology were in concordance with previous published description of Ilyonectria macrodidyma (Halleen, Schroers & Crous) P. Chaverri & C. Salgado (1,3,4). Identification to species level was confirmed by sequence comparison of four Californian isolates (UCCE958, UCCE959, UCCE960, and UCCE961) with sequences available in GenBank using the internal transcribed spacer region (ITS1-5.8S-ITS2) of the rDNA (primers ITS1/ITS4), a portion of the β-tubulin gene (BT1a/BT1b), and a partial sequence of the mitochondrial small subunit rDNA (NMS1/NMS2) (4). Fungal sequences of isolates from olive from California (GenBank JQ868543 to JQ868554) showed 99 to 100% homology with previously identified and deposited I. macrodidyma isolates in Genbank for all three genes. Pathogenicity of I. macrodidyma in olive cvs. Arbequina, Arbosan, and Koroneiki was investigated using two fungal isolates (UCCE958 and UCCE960) as reported by Petit and Gubler (4). The roots of 10 1-year-old trees per fungal isolate for each olive cultivar were individually inoculated with 25 ml of a 106 conidia/ml spore suspension and placed in a lath house at the UC Davis field station. Additionally, 10 trees per cultivar were inoculated with sterile water as controls. Six months after inoculation, most of the inoculated olive plants showed chlorotic leaves similar to those observed in commercial orchards. Root necrosis for each cv. was expressed as the percentage of root length having lesions (2). No significant difference was observed between isolates and average root necrosis was 29.4, 35.6, and 38.3% in Koroniki, Arbosana, and Arbequina, respectiveley. I. macrodidyma was recovered from symptomatic roots in each of the cvs. and identified based on morphology. No root rot symptoms were observed in the controls. To our knowledge, this is the first report of I. macrodidyma causing root rot of olive trees not only in California but anywhere in the world. References: (1) P. Chaverri et al. Stud. Mycol. 68:57, 2011. (2) M. Giovanetti and B. Mosse. New Phytol. 84:489, 1980. (3) F. Halleen et al. Stud. Mycol. 50:421, 2004. (4) E. Petit and W. D. Gubler. Plant Dis. 89:1051, 2005.


Genetic analysis of specific and redundant roles for p38alpha and p38beta MAPKs during mouse development.
p38α MAPK is an important regulator of cellular responses induced by external cues, but the elucidation of physiological functions for p38α has been complicated by the possible functional redundancy in vivo with the related family member p38β. We found that mice with combined deletion of p38α and p38β display diverse developmental defects at midgestation, including major cardiovascular abnormalities, which are observed neither in single knockout nor in double heterozygous embryos. Expression analysis indicates specific functions of p38α and p38β in the regulation of cardiac gene expression during development. By using knock-in animals that express p38β under control of the endogenous p38α promoter, we also found that p38β cannot perform all of the functions of p38α during embryogenesis. Our results identify essential roles for p38α and p38β during development and suggest that some specific functions may be explained by differences in expression patterns.


Ultrastructural observations on the fate of Brugia malayi in jirds previously vaccinated with irradiated infective stage larvae.
Vaccination of inbred jirds (Meriones unguiculatus) with 60cobalt radiation-attenuated Brugia malayi infective stage larvae (L3) protected against homologous challenge given either subcutaneously (sc) or by the intraperitoneal route (ip). At necropsy numerous nodules were recovered from the peritoneal cavities of jirds which had been vaccinated sc and challenged ip. Histopathologic analysis showed these to be granulomas containing dead and dying larvae and transmission electron microscopy showed that eosinophils were present in high numbers around and within the larvae. Structural damage to the L3 cuticle was apparent in discrete areas and eosinophils actively entering the breached cuticle at the time of fixation were observed. Coalescence of eosinophil secretion granules and the formation of degranulation vacuoles were seen in eosinophils throughout the granulomas. Degranulation resulted in the deposition of electron-dense material on the surface of the larval cuticle. The jird vaccine model for B. malayi thus appears to be a potentially useful tool for investigation of immune mechanisms in filariasis.


The "Mutated in Colorectal Cancer" Protein Is a Novel Target of the UV-Induced DNA Damage Checkpoint.
MCC is a potential tumor suppressor gene, which is silenced by promoter hypermethylation in a subset of colorectal cancers. However, its functions have remained poorly understood. In the present study, we describe a novel function of MCC in the DNA damage response. Several novel phosphorylation sites were identified by mass spectrometry, including 2 highly conserved ATM/ATR consensus sites at serine 118 and serine 120. In addition, exposure to ultraviolet radiation (UV), but not phleomycin, caused PI3K-dependent phosphorylation of MCC and its nuclear localization. Re-expression of MCC in HCT15 colorectal cancer cells led to a G2/M arrest, and MCC knockdown impaired the induction of a G2/M arrest following UV radiation. Finally, mutation of S118/120 to alanine did not affect MCC nuclear shuttling following UV but did impair MCC G2/M checkpoint activity. Thus, these results suggest that MCC is a novel target of the DNA damage checkpoint and that MCC is required for the complete cell cycle arrest in the G2/M phase in response to UV.


The simulated early learning of cervical spine manipulation technique utilising mannequins.
Trivial pain or minor soreness commonly follows neck manipulation and has been estimated at one in three treatments. In addition, rare catastrophic events can occur. Some of these incidents have been ascribed to poor technique where the neck is rotated too far. The aims of this study were to design an instrument to measure competency of neck manipulation in beginning students when using a simulation mannequin, and then examine the suitability of using a simulation mannequin to teach the early psychomotor skills for neck chiropractic manipulative therapy. We developed an initial set of questionnaire items and then used an expert panel to assess an instrument for neck manipulation competency among chiropractic students. The study sample comprised all 41 fourth year 2014 chiropractic students at Murdoch University. Students were randomly allocated into either a usual learning or mannequin group. All participants crossed over to undertake the alternative learning method after four weeks. A chi-square test was used to examine differences between groups in the proportion of students achieving an overall pass mark at baseline, four weeks, and eight weeks. This study was conducted between January and March 2014. We successfully developed an instrument of measurement to assess neck manipulation competency in chiropractic students. We then randomised 41 participants to first undertake either "usual learning" (n = 19) or "mannequin learning" (n = 22) for early neck manipulation training. There were no significant differences between groups in the overall pass rate at baseline (χ(2) = 0.10, p = 0.75), four weeks (χ(2) = 0.40, p = 0.53), and eight weeks (χ(2) = 0.07, p = 0.79). This study demonstrates that the use of a mannequin does not affect the manipulation competency grades of early learning students at short term follow up. Our findings have potentially important safety implications as the results indicate that students could initially gain competence in neck manipulation by using mannequins before proceeding to perform neck manipulation on each other.


First Report of Powdery Mildew Caused by Erysiphe arcuata on Lanceleaf Coreopsis (Coreopsis lanceolata) in Korea.
Lanceleaf coreopsis (Coreopsis lanceolata L.) is a plant species of the genus Coreopsis in Asteraceae native to the USA. This plant is a bushy perennial species with finely cut foliage and showy round flowers and is increasing as plants used in landscaping in the world. The invasive plant is also planted very commonly along roadsides in Korea. In late October 2011 and August 2012, signs and symptoms of a powdery mildew disease were observed on lanceleaf coreopsis in several land areas near Gwangju-river, Gwangju, Korea. Symptoms included typical white superficial mycelia. The conidia of the powdery mildew fungus occurred on adaxial and abaxial surfaces. Chasmothecia were not observed. Single conidia formed terminally on conidiophores. Conidial morphology was subcylindrical to oblong. Dimension of conidia was 23.6 to 41.4 (avg. 35.1) μm long × 11.3 to 18.2 (avg. 14.8) μm wide. Conidiophores were composed of five to six (up to seven) cells, ranged from 45.7 to 131.2 (avg. 98.1) μm long × 8.2 to 11.1 (avg. 8.3) μm wide with foot-cells straight to slightly flexuous. Oidium anamorph of this fungus matched that of E. arcuata U. Braun, Heluta and S. Takam. described by Pastircakova et al. (3). From extracted genomic DNA, the rDNA ITS was amplified with ITS1F (5'-CTTGGTCATTTAGAGGAAGT-3') and LR5F (5'-GCTATCCTGAGGGAAAC-3') primer set. The rDNA ITS homology of the fungus (EML-CDPW1, GenBank Accession No. JX485650) showed 100% (590/590) identity value with E. arcuata (GenBank Accession No. AB252459). The identification of the fungus as E. arcuata was based on morphological data combined with the results of sequence analysis. Until recently, E. arcuata has been known to widespread on Carpinus species of the family Betulaceae including European hornbeam (C. betulus L.) and Chonowski's hornbeam (C. tschonoskii Maxim.) in Asia and Europe since Braun et al. first reported it as a new species in 2006 (1). In Korea, Podosphaera fusca (= Sphaerotheca fusca) and P. fuliginea (= S. fuliginea) were reported to cause powdery mildews on Coreopsis lanceolata. E. cichoracearum (= Golovinomyces cichoracearum), Leveillula taurica, P. fusca (= S. fusca), and Oidium spp. have been reported on Coreopsis spp. in the world (3). To our knowledge, this is the first report of powdery mildew caused by Oidium anamorph of E. arcuata on lanceleaf coreopsis (C. lanceolata) in Korea or elsewhere in the world. References: (1) U. Braun et al. Mycol. Prog. 5:139, 2006. (2) D. F. Farr and A. Y. Rossman. Fungal Databases, Systematic Mycology and Microbiology Laboratory, ARS, USDA. http://nt.ars-grin.gov/fungaldatabases/ , 2012. (3) K. Pastircakova et al. J. Phytopathol. 156:597, 2008.


Modernizing mental health services for older people: a case study.
This paper describes an evaluation of a redevelopment program in a mental health service for older people, stimulated by U.K. Department of Health policy. IQCOL (Improving Quality of Care for Older People in Lambeth) was a two-year program to modernize and expand an inner-city service, with objectives to improve access, embed new functions, and tune the service towards the needs of the local community. The program evaluation aimed to contribute to knowledge on service planning and methodology for evaluating complex interventions. The study evaluated the progress and outcomes of this multifaceted program. The realist model of evaluation was followed, with a dual emphasis on utility and generalizability. With an iterative approach, the pragmatic, longitudinal design comprised a combination of qualitative and quantitative methods to explain the process of change and to measure achievement of objectives. A high level of participation in evaluation activities was achieved. The workforce generally responded well to the program. However, progress in one team was hindered by understaffing and resistance to change, emphasizing that while localized provision may be desirable, team viability requires adequate resources and professional support. Improved access was indicated by a 13% increase of referrals. Data suggested earlier referral of dementia cases. Carer support was implemented, but assertive outreach was impeded by professional boundary issues. Ethnicity data showed that the service was responding to demographic trends. Positive views towards the program were associated with team resources and recent professional training. This case study demonstrates how whole system change can be achieved if sufficient attention is given to the needs of staff implementing the program. The evaluation emphasizes the importance of context in producing generalizable evidence on service development, and contributes useful methodological insights.


PeptideManager: a peptide selection tool for targeted proteomic studies involving mixed samples from different species.
The search for clinically useful protein biomarkers using advanced mass spectrometry approaches represents a major focus in cancer research. However, the direct analysis of human samples may be challenging due to limited availability, the absence of appropriate control samples, or the large background variability observed in patient material. As an alternative approach, human tumors orthotopically implanted into a different species (xenografts) are clinically relevant models that have proven their utility in pre-clinical research. Patient derived xenografts for glioblastoma have been extensively characterized in our laboratory and have been shown to retain the characteristics of the parental tumor at the phenotypic and genetic level. Such models were also found to adequately mimic the behavior and treatment response of human tumors. The reproducibility of such xenograft models, the possibility to identify their host background and perform tumor-host interaction studies, are major advantages over the direct analysis of human samples. At the proteome level, the analysis of xenograft samples is challenged by the presence of proteins from two different species which, depending on tumor size, type or location, often appear at variable ratios. Any proteomics approach aimed at quantifying proteins within such samples must consider the identification of species specific peptides in order to avoid biases introduced by the host proteome. Here, we present an in-house methodology and tool developed to select peptides used as surrogates for protein candidates from a defined proteome (e.g., human) in a host proteome background (e.g., mouse, rat) suited for a mass spectrometry analysis. The tools presented here are applicable to any species specific proteome, provided a protein database is available. By linking the information from both proteomes, PeptideManager significantly facilitates and expedites the selection of peptides used as surrogates to analyze proteins of interest.


Effect of short-term mechanical stimulation on the salivary concentrations of vitamin C, vitamin E, total antioxidant capacity and total oxidant status.
Oxidative stress biomarkers of oral and systemic diseases can be found in saliva. However, there is no uniformity for the saliva collection time in these kinds of analyses and saliva composition may change because of mechanical stimulation. Therefore, the aim of this work was to determine the effect of mechanical stimulation for 10 min on the concentrations of vitamin C, vitamin E, total antioxidants and total oxidants in saliva. Saliva samples from individuals of both sexes, aged between 18 and 38 years, were collected for 10 min at 2 minintervals. Saliva flow rate in each 2 min period was measured, as well the total oxidant state, the total antioxidant capacity, vitamin C and vitamin E concentrations. All analyses were performed in triplicate and were determined using colorimetric tests. The results were analysed using t-test, Friedman's test and analysis of variance (ANOVA) for repeated measures. Mauchly's sphericity test was applied and, if necessary, technical corrections were made using the Greenhouse-Geisser test. We found no significant difference between the amounts of saliva produced across the collection times. Total oxidant status, total antioxidant capacity, vitamin C and vitamin E concentrations remained stable. Based on our findings, saliva can be collected for 10 min or less with masticatory stimulation without any variations in the concentration of the variables analysed. However, we suggest using saliva samples after two minutes of mechanical stimulation.


Day/night whole sky imagers for 24-h cloud and sky assessment: history and overview.
A family of fully automated digital whole sky imagers (WSIs) has been developed at the Marine Physical Laboratory over many years, for a variety of research and military applications. The most advanced of these, the day/night whole sky imagers (D/N WSIs), acquire digital imagery of the full sky down to the horizon under all conditions from full sunlight to starlight. Cloud algorithms process the imagery to automatically detect the locations of cloud for both day and night. The instruments can provide absolute radiance distribution over the full radiance range from starlight through daylight. The WSIs were fielded in 1984, followed by the D/N WSIs in 1992. These many years of experience and development have resulted in very capable instruments and algorithms that remain unique. This article discusses the history of the development of the D/N WSIs, system design, algorithms, and data products. The paper cites many reports with more detailed technical documentation. Further details of calibration, day and night algorithms, and cloud free line-of-sight results will be discussed in future articles.


Identification of larval-stage-specific antigens of Onchocerca volvulus uniquely recognized by putative immune sera from humans and vaccination sera from animal models.
Immunity to Onchocerca volvulus is indicated by the existence of putative immune individuals (PI), who do not develop patent infections or clinical symptoms after living in endemic areas for extended periods of time. To gain insight into the nature of the humoral response of PI from Liberia and Ecuador, their sera and those of infected individuals were investigated and compared using western blots. The antigen preparations used were extracts of third- and fourth-stage larvae (L3 and L4, respectively), excretory-secretory proteins (ES-L3) and extracts of third-stage larvae (L3-day2) that had been cultured for 2 days, and extracts of nodular and skin microfilariae. Whereas some antigens were commonly recognized by all individuals, many stage-specific antigens (of 68, 60, 55, 46, 43, 42, 20 and 18 kDa in L3; 100, 88, 80, 50, 48, 45, 40 and 20 kDa in L3-day2; 110, 80 and 72 kDa in ES-L3; 48, 18, 16 and 14 kDa in L4; and 115, 60, 47, 43 and 17 kDa in skin and/or nodular microfilariae) were uniquely recognized by sera from PI. Some of the unique antigens were also recognized by sera from mice and a chimpanzee that were resistant to challenge after immunization with irradiated third-stage larvae. The results support the validity of the assumption that a unique status of immunity develops in certain individuals living in areas where onchocerciasis is endemic.


Participation as an Outcome in Multiple Sclerosis Falls-Prevention Research: Consensus Recommendation from the International MS Falls Prevention Research Network.
Selecting the outcomes for an intervention trial is a key decision that influences many other aspects of the study design. One of the major tasks during the 3-day inaugural meeting of the International MS Falls Prevention Research Network was to identify the key outcomes for the falls-prevention intervention that was being designed by the Network members for testing across their multiple sites. Through a nominal group process, meeting participants described how engagement in important, meaningful everyday activities, beyond traditional basic and instrumental activities of daily living, should be a long-term outcome of a successful falls-prevention intervention for people with MS. Post-meeting work, which involved literature reviews and comparisons of definitions of major constructs identified during the meeting discussions, led to the consensus recommendation of including participation as a long-term outcome in MS falls-prevention interventions. Participation reflects involvement in a life situation. This article explains the rationale for this recommendation and presents four measures that have the potential for use in tracking long-term participation outcomes in MS falls-prevention research.


Detection of aberrant hippocampal mossy fiber connections: Ex vivo mesoscale diffusion MRI and microtractography with histological validation in a patient with uncontrolled temporal lobe epilepsy.
Understanding the neurobiology and functional connectivity of hippocampal structures is essential for improving the treatment of mesial temporal lobe epilepsy. At the macroscale, in vivo MRI often reveals hippocampal atrophy and decreased fractional anisotropy, whereas at the microscopic scale, there frequently is evidence of neuronal loss and gliosis. Mossy fiber sprouting in the dentate gyrus (DG), with evidence of glutamatergic synapses in the stratum moleculare (SM) putatively originating from granule cell neurons, may also be observed. This aberrant connection between the DG and SM could produce a reverberant excitatory circuit. However, this hypothesis cannot easily be evaluated using macroscopic or microscopic techniques. We here demonstrate that the ex vivo mesoscopic MRI of surgically excised hippocampi can bridge the explanatory and analytical gap between the macro- and microscopic scale. Specifically, diffusion- and T2 -weighted MRI can be integrated to visualize a cytoarchitecture that is akin to immunohistochemistry. An appropriate spatial resolution to discern individual cell layers can then be established. Processing of diffusion tensor images using tractography detects extra- and intrahippocampal connections, hence providing a unique systems view of the hippocampus and its connected regions. Here, this approach suggests that there is indeed an aberrant connection between the DG and SM, supporting the sprouting hypothesis of a reverberant excitatory network. Mesoscopic ex vivo MR imaging hence provides an exciting new avenue to study hippocampi from treatment-resistant patients and allows exploration of existing hypotheses, as well as the development of new treatment strategies based on these novel insights. Hum Brain Mapp 37:780-795, 2016. © 2015 Wiley Periodicals, Inc.


Gynecologic cancers in pregnancy: guidelines of a second international consensus meeting.
This study aimed to provide timely and effective guidance for pregnant women and health care providers to optimize maternal treatment and fetal protection and to promote effective management of the mother, fetus, and neonate when administering potentially teratogenic medications. New insights and more experience were gained since the first consensus meeting 5 years ago. Members of the European Society of Gynecological Oncology task force "Cancer in Pregnancy" in concert with other international experts reviewed the existing literature on their respective areas of expertise. The summaries were subsequently merged into a complete article that served as a basis for discussion during the consensus meeting. All participants approved the final article. In the experts' view, cancer can be successfully treated during pregnancy in collaboration with a multidisciplinary team, optimizing maternal treatment while considering fetal safety. To maximize the maternal outcome, cancer treatment should follow a standard treatment protocol as for nonpregnant patients. Iatrogenic prematurity should be avoided. Individualization of treatment and effective psychologic support is imperative to provide throughout the pregnancy period. Diagnostic procedures, including staging examinations and imaging, such as magnetic resonance imaging and sonography, are preferable. Pelvic surgery, either open or laparoscopic, as part of a treatment protocol, may reveal beneficial outcomes and is preferably performed by experts. Most standard regimens of chemotherapy can be administered from 14 weeks gestational age onward. Apart from cervical and vulvar cancer, as well as important vulvar scarring, the mode of delivery is determined by the obstetrician. Term delivery is aimed for. Breast-feeding should be considered based on individual drug safety and neonatologist-breast-feeding expert's consult. Despite limited evidence-based information, cancer treatment during pregnancy can succeed. State-of-the-art treatment should be provided for this vulnerable population to preserve maternal and fetal prognosis. Supplementary data on teratogenic effects, ionizing examinations, sentinel lymph node biopsy, tumor markers during pregnancy, as well as additional references and tables are available at the extended online version of this consensus article, go to http://links.lww.com/IGC/A197.


Responses of group III and IV muscle afferents to dynamic exercise.
Tetanic contraction of hindlimb skeletal muscle, induced by electrical stimulation of either ventral roots or peripheral nerves, is well known to activate group III and IV afferents. Nevertheless, the effect of dynamic exercise on the discharge of these thin fiber afferents is unknown. To shed some light on this question, we recorded in decerebrate cats the discharge of 24 group III and 10 group IV afferents while the mesencephalic locomotor region (MLR) was stimulated electrically. Each of the 34 afferents had their receptive fields in the triceps surae muscles. Stimulation of the MLR for 1 min caused the triceps surae muscles to contract rhythmically, an effect induced by an alpha-motoneuron discharge pattern and recruitment order almost identical to that occurring during dynamic exercise. Eighteen of the 24 group III and 8 of the 10 group IV muscle afferents were stimulated by MLR stimulation. The oxygen consumption of the dynamically exercising triceps surae muscles was increased by 2.5-fold over their resting levels. We conclude that low levels of dynamic exercise stimulate group III and IV muscle afferents.


Natural variants of ELF3 affect thermomorphogenesis by transcriptionally modulating PIF4-dependent auxin response genes.
Perception and transduction of temperature changes result in altered growth enabling plants to adapt to increased ambient temperature. While PHYTOCHROME-INTERACTING FACTOR4 (PIF4) has been identified as a major ambient temperature signaling hub, its upstream regulation seems complex and is poorly understood. Here, we exploited natural variation for thermo-responsive growth in Arabidopsis thaliana using quantitative trait locus (QTL) analysis. We identified GIRAFFE2.1, a major QTL explaining ~18 % of the phenotypic variation for temperature-induced hypocotyl elongation in the Bay-0 x Sha recombinant inbred line population. Transgenic complementation demonstrated that allelic variation in the circadian clock regulator EARLY FLOWERING3 (ELF3) is underlying this QTL. The source of variation could be allocated to a single nucleotide polymorphism in the ELF3 coding region, resulting in differential expression of PIF4 and its target genes, likely causing the observed natural variation in thermo-responsive growth. In combination with other recent studies, this work establishes the role of ELF3 in the ambient temperature signaling network. Natural variation of ELF3-mediated gating of PIF4 expression during nightly growing periods seems to be affected by a coding sequence quantitative trait nucleotide that confers a selective advantage in certain environments. In addition, natural ELF3 alleles seem to differentially integrate temperature and photoperiod information to induce architectural changes. Thus, ELF3 emerges as an essential coordinator of growth and development in response to diverse environmental cues and implicates ELF3 as an important target of adaptation.


Dynamic transitions of the transmembrane domain of diphtheria toxin: disulfide trapping and fluorescence proximity studies.
Translocation of the catalytic domain of diphtheria toxin across the endosomal membrane to the cytosolic compartment depends on low-pH-triggered insertion of the toxin's T (transmembrane) domain into the membrane. The T domain, consisting of nine alpha-helices arranged in three layers, was cloned and expressed as a discrete protein in Escherichia coli, and mutant forms were prepared and characterized. To investigate the relative movements of the three layers under various conditions, we generated two mutant forms of the domain, each containing an artificial intramolecular disulfide bridge linking the buried apolar hairpin (TH8-TH9) to one of the other two layers. Both disulfides inhibited exposure of the domain's apolar regions in solution at low pH, as determined by 2-p-toluidinylnaphthalene-6-sulfonate binding, and blocked its ability to form channels in artificial bilayers. Reduction of the bridges abolished these effects. Reduced forms of the mutant proteins were reacted with pyrenylmaleimide, a fluorescent probe, to monitor separation of the layers. Strong excimer bands seen in both mutants at neutral pH were undiminished at pH 5, indicating the retention of gross conformation in solution under acidic conditions. The addition of phospholipid vesicles at pH 5, but not at pH 7.5, quenched excimer fluorescence, reflecting the physical separation of the TH8-TH9 hairpin from the other layers upon the T domain's interaction with the bilayer. The results indicate that (i) the conformation of the isolated T domain closely resembles that seen in the whole toxin, (ii) the TH8-TH9 hairpin separates from both of the other layers of the domain as an essential step of membrane insertion, and (iii) this separation is triggered by contact of the domain with the membrane under acidic conditions.


High efficiency quantum dot light emitting diodes from positive aging.
Colloidal quantum dot-polymer hybrid light emitting diodes (QLEDs) that exhibit external quantum efficiencies >12% for all three primary colors (21% from green) have been demonstrated. These high efficiencies result in part from a positive aging effect reported here for the first time, where positive aging means the efficiency of the QLED increased with time. We have achieved 470 h operational life time (T90) at 2550 nits for red QLEDs. At longer times, negative aging phenomena lead to lower luminance and limit the lifetime of the QLEDs. It is concluded that we have reasonable control over the efficiency of QLEDs. The next challenge is to achieve lifetimes sufficiently long for all three primary colors for applications such as in television and illumination.


Postnatal microbial colonization programs the hypothalamic-pituitary-adrenal system for stress response in mice.
Indigenous microbiota have several beneficial effects on host physiological functions; however, little is known about whether or not postnatal microbial colonization can affect the development of brain plasticity and a subsequent physiological system response. To test the idea that such microbes may affect the development of neural systems that govern the endocrine response to stress, we investigated hypothalamic-pituitary-adrenal (HPA) reaction to stress by comparing germfree (GF), specific pathogen free (SPF) and gnotobiotic mice. Plasma ACTH and corticosterone elevation in response to restraint stress was substantially higher in GF mice than in SPF mice, but not in response to stimulation with ether. Moreover, GF mice also exhibited reduced brain-derived neurotrophic factor expression levels in the cortex and hippocampus relative to SPF mice. The exaggerated HPA stress response by GF mice was reversed by reconstitution with Bifidobacterium infantis. In contrast, monoassociation with enteropathogenic Escherichia coli, but not with its mutant strain devoid of the translocated intimin receptor gene, enhanced the response to stress. Importantly, the enhanced HPA response of GF mice was partly corrected by reconstitution with SPF faeces at an early stage, but not by any reconstitution exerted at a later stage, which therefore indicates that exposure to microbes at an early developmental stage is required for the HPA system to become fully susceptible to inhibitory neural regulation. These results suggest that commensal microbiota can affect the postnatal development of the HPA stress response in mice.


[Yearly changes in antibacterial activities of cefozopran against various clinical isolates between 1996 and 2001--I. Gram-positive bacteria].
The in vitro antibacterial activities of cefozopran (CZOP), an agent of cephems, against various clinical isolates obtained between 1996 and 2001 were yearly evaluated and compared with those of other cephems, oxacephems, carbapenems, and penicillins. A total of 1,274 strains in 15 species of Gram-positive bacteria were isolated from the clinical materials annually collected from January to December, and consisted of methicillin-susceptible Staphylococcus aureus (MSSA), methicillin-resistant Staphylococcus aureus (MRSA), Staphylococcus epidermidis, Staphylococcus haemolyticus, Streptococcus pyogenes, Streptococcus agalactiae, Streptococcus pneumoniae, Enterococcus faecalis, Enterococcus faecium, Enterococcus avium, and Peptostreptococcus spp. CZOP possessed stable antibacterial activities against all strains tested throughout 6 years. The MIC90 of CZOP against MRSA and S. haemolyticus tended to decrease while against S. pneumoniae and Peptostreptococcus spp., tended to increase year by year. However, the MIC90 just changed a little and were consistent with the results from the studies performed until the new drug application approval. Increases in the MIC90 against S. pneumoniae were also observed with ceftazidime (CPR), cefepime (CFPM), flomoxef (FMOX), sulbactam/cefoperazone (SBT/CPZ), and imipenem (IPM). Increases in the MIC90 against Peptostreptococcus spp. were also observed with FMOX, SBT/CPZ, and IPM. In conclusion, the annual antibacterial activities of CZOP against the Gram-positive bacteria did not considerably change. It, therefore, was suggested that CZOP had maintained high antibacterial activity during 6 years of post marketing.


What are the Facilitators and Obstacles to Participation in Workplace Team Sport? A Qualitative Study.
Working age adults are failing to meet physical activity recommendations. Inactive behaviours are increasing costs for diminished individual and organisational health. The workplace is a priority setting to promote physical activity, however there is a lack of evidence about why some employees choose to participate in novel workplace activities, such as team sport, whilst others do not. The aim of this study was to explore the complexity of facilitators and obstacles associated with participation in workplace team sport. Twenty-nine semi-structured face-to-face and telephone interviews were conducted with office workers (58% female) (36 ± 7.71) from manufacturing, public services, and educational services. Data was analysed through template analysis. Five sub-level (i.e., intrapersonal, interpersonal, organisational, community and societal influences) facilitate participation or create obstacles for participants. Participants were challenged by a lack of competence, self-efficacy, negative sporting ideals and amotivation. Unhealthy competition, an unstable work-life balance and unsupportive colleagues created obstacles to participation. An unsupportive organisation and workplace culture placed demands on workplace champions, funding, facilities and communication. Healthy competitions, high perceptions of competence and self-efficacy, and being motivated autonomously enabled participation. Further, relatedness and social support created a physical activity culture where flexible working was encouraged and team sport was promoted in accessible locations within the organisation. Researchers should consider accounting for complexity of these influences. A participatory approach may tailor interventions to individual organisations and the employees that work within them. Interventions whereby autonomy, competence and relatedness are supported are recommended. This may be achieved by adapting sports and training workplace champions.


Decalcification of Breast Cancer Bone Metastases With EDTA Does Not Affect ER, PR, and HER2 Results.
In metastatic breast cancer (MBC), expression of estrogen receptor (ER), progesterone receptor (PR), and human epidermal growth factor receptor-2 (HER2) guides treatment selection. In case of bone-only metastatic disease, ER, PR, and HER2 status assessment may be hampered by decalcification. We aimed to determine the optimal decalcification method, and to study discordance of receptor expression between paired primary breast tumors and optimally decalcified bone metastases. First, decalcification was simulated using acetic acid, hydrochloric/formic acid, and EDTA on 12 primary breast carcinomas. ER, PR, and HER2 immunohistochemistry (IHC) and HER2 in situ hybridization (ISH) were assessed, before and after the 3 decalcification methods. EDTA was considered the optimal method, as it did not affect IHC and as ISH failed in only 1/16 cases. Hydrochloric/formic acid altered ER and PR results, and, with acetic acid and hydrochloric/formic acid, ISH failed in, respectively, 94% and 100%. Second, ER, PR, and HER2 IHC was performed in paired primary tumors and EDTA-decalcified bone metastases obtained from patients with first presentation of MBC. Clinically relevant discordance was defined as changed receptor status with treatment implications. Paired samples of 77 patients, participating in the IMPACT-MBC trial, were evaluable. Hormonal receptor expression change was clinically relevant in 6 patients (7.9%) and HER2 expression change in 1 patient (1.3%). This study shows that EDTA decalcification minimally affects receptor expression results. The incidence of clinically relevant discordance between the primary tumor and bone metastases is low. These findings support that bone biopsies can reliably be used to assess receptor status.