Expression of E-cadherin and beta-catenin in human non-small cell lung cancer and the clinical significance.
E-cadherin, a calcium-dependent cell-cell adhesion molecule, plays a key role in the maintenance of tissue integrity. The function of this molecule is partly mediated by alpha-/beta-/gamma-catenin. Loss or dysfunction of E-cadherin is associated with an invasive phenotype. We analyzed the expression of E-cadherin and beta-catenin in human lung cancer to determine the relationship to clinicopathological factors and prognosis. E-cadherin and beta-catenin expressions were evaluated in 331 lung cancer tissues in a immunohistochemical analysis. Reduced E-cadherin expression was evident in 138 (42%), and reduced beta-catenin expression was noted in 122 (37%). Reduced E-cadherin expression significantly correlated with lymph nodes metastasis (P = 0.0199). E-cadherin expression significantly correlated with increasing histological differentiation (P = 0.0403). Although reduced E-cadherin did not correlate with the prognosis (P = 0.0652), reduced beta-catenin expression did significantly correlate with a poor prognosis (P = 0.0001). When both were reduced, there was a significant unfavorable prognosis compared with either the reduced expression (P = 0.0493) and preserved expression (P = 0.0003). Multivariate analysis showed a significantly lower survival rate for patients with reduced beta-catenin (P < 0.0001). We interpret these data to mean that dysfunction of the cell-cell adhesion molecule has a role in the progression of lung cancer and that analysis of E-cadherin and beta-catenin expression can provide clinically important evidence on which to base treatment.


Removal of lymphangiectasis using high-frequency radio wave electrosurgery.
To describe a novel technique using high-frequency radio wave electrosurgery (Ellman Surgitron Dual Frequency RF; Ellman International, Inc) for the management of lymphangiectasis. Ablations were performed at the lowest power setting of 1 (of 100) in the cutting mode, producing the least amount of lateral heat. To prevent conjunctival hemorrhage, ablation was initiated immediately before introducing the tip of a needle electrode into the subconjunctival tissue under the target area. After the tip of the electrode reached the target area, ablation was maintained for 1 to 2 seconds on the surrounding area and for a longer time on the dilated lymphatic vessels, until the target conjunctiva blanched. These ablations were performed cautiously with the fine electrode to avoid thermal injury to the adjacent scleral tissue. Persistent lymphangiectasis accompanied by accumulated fluid was successively treated with high-frequency radio wave electrosurgery. Surgical time was less than 5 minutes. There were no intraoperative complications. Fourier domain optical coherence tomography revealed resolution of the accumulated fluid and a decrease of dilated lymphatic vessels. Postoperatively, no notable complications, such as charring, scarring of Tenon capsule, or symblepharon resulting from excessive cauterization, were observed. High-frequency radio wave electrosurgery may be a safe, quick, and effective modality for the treatment of symptomatic lymphangiectasis patients.


Seroprevalence and correlates of human papillomavirus 16/18 seropositivity among young women in Costa Rica.
Serological indicators of human papillomavirus (HPV) infection are being used to differentiate HPV-naïve from previously infected women in vaccine and epidemiologic/clinical studies. We investigated HPV16 and 18 seroepidemiology among young, unvaccinated women aged between 18 and 25. We conducted a cross-sectional evaluation of the enrollment visit in the ongoing community-based HPV16/18 Costa Rica Vaccine Trial. Prevaccination serum immunoglobulin G (IgG) antibodies were measured against HPV16 and HPV18 by enzyme-linked immunosorbent assay; cervical samples were tested for HPV DNA using Hybrid Capture 2 and SPF10/LiPA25. Seroprevalence and its correlates were evaluated using unconditional logistic regression. Among 5871 nonvirginal women, HPV16 and 18 seroprevalences were 30.8% and 28.1%, HPV16 and HPV18 DNA prevalences were 8.3% and 3.2%, respectively. About 37% of HPV16 DNA-positives and 42% of HPV18 DNA-positives were seronegative. Seroprevalence increased with time since sexual debut, whereas DNA prevalence did not. The correlates of HPV16 and/or 18 seropositivity were related to sexual behaviors, particularly higher number of lifetime sexual partners. There was no evidence of assay cross-reactivity as HPV16 seroprevalence was similar (approximately 34%) among women singly infected with genetically and nongenetically related species (α9 and non-α9); likewise, seropositivity to HPV18 was similar (approximately 30%) among women singly infected with α7 and non-α7 species. The increasing seroprevalence observed with time since first sex suggests that HPV serology is a cumulative marker of HPV exposure. However, many DNA infected women were seronegative; thus, serology is an imperfect measure of past exposure to cervical HPV, at best. Additionally, we found no evidence of assay cross-reactivity.


Scale-dependent response diversity of seabirds to prey in the North Sea.
Functional response diversity is defined as the diversity of responses to environmental change among species that contribute to the same ecosystem function. Because different ecological processes dominate on different spatial and temporal scales, response diversity is likely to be scale dependent. Using three extensive data sets on seabirds, pelagic fish, and zooplankton, we investigate the strength and diversity in the response of seabirds to prey in the North Sea over three scales of ecological organization. Two-stage analyses were used to partition the variance in the abundance of predators and prey among the different scales of investigation: variation from year to year, variation among habitats, and variation on the local patch scale. On the year-to-year scale, we found a strong and synchronous response of seabirds to the abundance of prey, resulting in low response diversity. Conversely, as different seabird species were found in habitats dominated by different prey species, we found a high diversity in the response of seabirds to prey on the habitat scale. Finally, on the local patch scale, seabirds were organized in multispecies patches. These patches were weakly associated with patches of prey, resulting in a weak response strength and a low response diversity. We suggest that ecological similarities among seabird species resulted in low response diversity on the year-to-year scale. On the habitat scale, we suggest that high response diversity was due to interspecific competition and niche segregation among seabird species. On the local patch scale, we suggest that facilitation with respect to the detection and accessibility of prey patches resulted in overlapping distribution of seabirds but weak associations with prey. The observed scale dependencies in response strength and diversity have implications for how the seabird community will respond to different environmental disturbances.


M13 bacteriophage spheroids as scaffolds for directed synthesis of spiky gold nanostructures.
The spherical form (s-form) of a genetically-modified gold-binding M13 bacteriophage was investigated as a scaffold for gold synthesis. Repeated mixing of the phage with chloroform caused a 15-fold contraction from a nearly one micron long filament to an approximately 60 nm diameter spheroid. The geometry of the viral template and the helicity of its major coat protein were monitored throughout the transformation process using electron microscopy and circular dichroism spectroscopy, respectively. The transformed virus, which retained both its gold-binding and mineralization properties, was used to assemble gold colloid clusters and synthesize gold nanostructures. Spheroid-templated gold synthesis products differed in morphology from filament-templated ones. Spike-like structures protruded from the spherical template while isotropic particles developed on the filamentous template. Using inductively coupled plasma-mass spectroscopy (ICP-MS), gold ion adsorption was found to be comparatively high for the gold-binding M13 spheroid, and likely contributed to the dissimilar gold morphology. Template contraction was believed to modify the density, as well as the avidity of gold-binding peptides on the scaffold surface. The use of the s-form of the M13 bacteriophage significantly expands the templating capabilities of this viral platform and introduces the potential for further morphological control of a variety of inorganic material systems.


Endomyocardial biopsy: a review of the literature.
A review of the literature relating to endomyocardial biopsy (EMB) is presented. This is considered important at this time since EMB is being utilized with increasing frequency, particularly for the diagnosis of myocarditis. The development of the technique is briefly outlined. Emphasis is placed on the clinical application of EMB in the various primary cardiomyopathies (dilated, hypertrophic, restrictive, and obliterative), the infiltrative secondary cardiomyopathies (amyloidosis, sarcoidosis, hemochromatosis), myocarditis, as well as such conditions as adriamycin cardiotoxicity, cardiac transplant rejection, and Kawasaki disease. More controversial application of EMB in primary mitral valve prolapse (Barlow's syndrome), idiopathic ventricular arrhythmias, and the elucidation of the enigmatic finding of angina with angiographically normal coronary arteries is detailed. Experience with immunological and biochemical investigation of biopsy material, as well as with virus isolation and drug assays in the myocardium, is alluded to. Complications encountered with this procedure are also discussed, and its future role is contemplated.


Clinical assessment of three common tests for traumatic anterior shoulder instability.
Although traumatic anterior shoulder instability is common, the usefulness of various physical examination tests as tools for the diagnosis of this condition has been studied infrequently. We hypothesized that (1) such tests would be specific but not sensitive for this condition, (2) the usefulness of the anterior drawer test would be limited because of pain during the test, and (3) an anterior drawer test would be a useful adjunct for making the diagnosis if it reproduced the instability symptoms. Between 2000 and 2004, 363 patients underwent a physical examination followed by shoulder arthroscopy. Forty-six patients with traumatic anterior shoulder instability that had been noted arthroscopically or documented radiographically after the trauma were included in our study group, and the remaining patients served as controls. The clinical usefulness of three tests (anterior apprehension, relocation, and anterior drawer tests) performed during the physical examination to make a diagnosis of traumatic anterior instability then was evaluated with statistical methods to assess their sensitivity, specificity, and likelihood ratios. If demonstration (or relief) of apprehension was used as the diagnostic criterion for a positive test, the sensitivity, specificity, and likelihood ratio were 72%, 96%, and 20.2, respectively, for the apprehension test and 81%, 92%, and 10.4, respectively, for the relocation test. If pain (or relief of pain) was used as the diagnostic criterion for a positive test, the values for the sensitivity, specificity, and likelihood ratio of both tests were lower. The anterior drawer test could be performed successfully in the physician's office for 87% of the patients. If reproduction of instability symptoms was used as the criterion for a positive anterior drawer test, the sensitivity, specificity, and likelihood ratio values of that test were 53%, 85%, and 3.6, respectively. The three physical examination tests for traumatic anterior shoulder instability are specific but not sensitive. Apprehension is a better criterion than pain for a positive apprehension or relocation test. The anterior drawer test (when pain does not prevent it from being performed) is helpful for diagnosing traumatic anterior instability. Diagnostic Level I. See Instructions to Authors for a complete description of levels of evidence.


Length-dependent optical effects in single walled carbon nanotubes.
Recently, Heller et al. reported length-dependent effects on the relative photoluminescence (PL) quantum yield of single walled carbon nanotubes (SWNTs) [Heller et al J. Am. Chem. Soc. 2004, 126, 14567-14573]. We propose a simple model involving thermal diffusion of excitons along the nanotube axis and quenching at the ends, to explain the observed trend in their data. By fitting to our model, we extract a diffusion coefficient of 6 cm(2)/s for excitons in SWNTs. Assuming a mono exponential decay of exciton PL, we also predict that effective length-dependent PL lifetimes for these excitons lie in the range of 1-27 ps. Experimental observations are shown to be consistent with stochastic rather than wavepacket-like exciton migration, which is in agreement with ultrafast excitonic dephasing. Edge effects seem to limit the use of short SWNTs in imaging and optical sensing applications.


Characterizing Population EEG Dynamics throughout Adulthood.
For decades, electroencephalography (EEG) has been a useful tool for investigating the neural mechanisms underlying human psychological processes. However, the amount of time needed to gather EEG data means that most laboratory studies use relatively small sample sizes. Using the Muse, a portable and wireless four-channel EEG headband, we obtained EEG recordings from 6029 subjects 18-88 years in age while they completed a category exemplar task followed by a meditation exercise. Here, we report age-related changes in EEG power at a fine chronological scale for δ, θ, α, and β bands, as well as peak α frequency and α asymmetry measures for both frontal and temporoparietal sites. We found that EEG power changed as a function of age, and that the age-related changes depended on sex and frequency band. We found an overall age-related shift in band power from lower to higher frequencies, especially for females. We also found a gradual, year-by-year slowing of the peak α frequency with increasing age. Finally, our analysis of α asymmetry revealed greater relative right frontal activity. Our results replicate several previous age- and sex-related findings and show how some previously observed changes during childhood extend throughout the lifespan. Unlike previous age-related EEG studies that were limited by sample size and restricted age ranges, our work highlights the advantage of using large, representative samples to address questions about developmental brain changes. We discuss our findings in terms of their relevance to attentional processes and brain-based models of emotional well-being and aging.


Parenting Predictors of Delay Inhibition in Socioeconomically Disadvantaged Preschoolers.
This study examined longitudinal associations between specific parenting factors and delay inhibition in socioeconomically disadvantaged preschoolers. At Time 1, parents and 2- to 4-year-old children (mean age = 3.21 years; N = 247) participated in a videotaped parent-child free play session, and children completed delay inhibition tasks (gift delay-wrap, gift delay-bow, and snack delay tasks). Three months later, at Time 2, children completed the same set of tasks. Parental responsiveness was coded from the parent-child free play sessions, and parental directive language was coded from transcripts of a subset of 127 of these sessions. Structural equation modeling was used, and covariates included age, gender, language skills, parental education, and Time 1 delay inhibition. Results indicated that in separate models, Time 1 parental directive language was significantly negatively associated with Time 2 delay inhibition, and Time 1 parental responsiveness was significantly positively associated with Time 2 delay inhibition. When these parenting factors were entered simultaneously, Time 1 parental directive language significantly predicted Time 2 delay inhibition whereas Time 1 parental responsiveness was no longer significant. Findings suggest that parental language that modulates the amount of autonomy allotted the child may be an important predictor of early delay inhibition skills.


Synaptic strength as a function of motor unit size in the normal frog sartorius.
A wide range of motor unit sizes exists in each frog sartorius, with values of tetanus and twitch tensions extending over 62- and 400-fold ranges, respectively. These differences primarily represent differences in the number, rather than the size, of muscle fibres in each motor unit. Tetanus-to-twitch tension ratios varied markedly between different motor units, ranging from 1.5 to over 190. Because directly stimulated muscle fibres have tetanus/twitch ratios no larger than 3, it is concluded that high tetanus/twitch ratios arise from recruitment of muscle fibres during the tetanus which respond with only subthreshold depolarization during a twitch. Small motor units, as judged either by their twitch or tetanus tension, were associated with higher tetanus/twitch ratios, suggesting that the average safety margin of a motor unit increases with the motor unit's size. Indeed, when the tetanus/twitch ratio of a motor unit is used to determine the fraction of fibres in the unit with subthreshold neuromuscular junctions, it is observed that there is a direct linear relationship between the size of the motor unit and its over-all efficacy of synaptic transmission. Measurements of the effects of changes in calcium concentration on motor unit twitch tension confirmed the last conclusion. Furthermore, this analysis revealed that large motor units, although having a wide range of transmission safety margins, are largely comprised of junctions of uniformly high safety margin. In motor units of smaller size, synaptic strengths become more evenly distributed over a wide range of values. Small motor units had consistently longer twitch rise-times than did larger units. This decrease in rise-time with motor unit size paralleled the decrease in tetanus/twitch ratios, raising questions about the regulation of muscle fibre contraction kinetics.


Botulinum toxin type A reduces pain supersensitivity in experimental diabetic neuropathy: bilateral effect after unilateral injection.
We investigated antinociceptive activity of botulinum toxin type A (BTX-A) in a model of diabetic neuropathic pain in rats. Male Wistar rats were made diabetic by a single intraperitoneal injection of streptozotocin (80mg/kg). Sensitivity to mechanical and thermal stimuli was measured with the paw-pressure and hot-plate test, respectively. The formalin test was used to measure sensitivity to chemical stimuli. Diabetic animals with pain thresholds lower for at least 25% compared to the non-diabetic group were considered neuropathic and were injected with BTX-A either subcutaneously (3, 5 and 7U/kg) or intrathecally (1U/kg). Mechanical and thermal sensitivity was measured at several time-points. After peripheral application, BTX-A (5 and 7U/kg) reduced mechanical and thermal hypersensitivity not only on ipsilateral, but on contralateral side, too. The antinociceptive effect started 5days following BTX-A injection and lasted at least 15days. Formalin-induced hypersensitivity in diabetic animals was abolished as well. When applied intrathecally, BTX-A (1U/kg) reduced diabetic hyperalgesia within 24h supporting the assumption of retrograde axonal transport of BTX-A from the peripheral site of injection to central nervous system. The results presented here demonstrate the long-lasting pain reduction after single BTX-A injection in the animals with diabetic neuropathy. The bilateral pain reduction after unilateral toxin application and the effectiveness of lower dose with the faster onset after the intrathecal injection suggest the involvement of the central nervous system in the antinociceptive action of BTX-A in painful diabetic neuropathy.


Statistical characterization of multiple-reaction monitoring mass spectrometry (MRM-MS) assays for quantitative proteomics.
Multiple reaction monitoring mass spectrometry (MRM-MS) with stable isotope dilution (SID) is increasingly becoming a widely accepted assay for the quantification of proteins and peptides. These assays have shown great promise in relatively high throughput verification of candidate biomarkers. While the use of MRM-MS assays is well established in the small molecule realm, their introduction and use in proteomics is relatively recent. As such, statistical and computational methods for the analysis of MRM-MS data from proteins and peptides are still being developed. Based on our extensive experience with analyzing a wide range of SID-MRM-MS data, we set forth a methodology for analysis that encompasses significant aspects ranging from data quality assessment, assay characterization including calibration curves, limits of detection (LOD) and quantification (LOQ), and measurement of intra- and interlaboratory precision. We draw upon publicly available seminal datasets to illustrate our methods and algorithms.


Overexpression of a dominant-negative allele of YPT1 inhibits growth and aspartyl protease secretion in Candida albicans.
To investigate the pre-Golgi secretion pathway in the pathogenic yeast Candida albicans, we cloned the C. albicans homologue of the Saccharomyces cerevisiae protein secretion gene YPT1. The C. albicans YPT1 ORF contained a 624 bp intronless ORF encoding a deduced protein of 207 aa and 2.3 kDa. This deduced protein was 77% identical to S. cerevisiae Ypt1 protein (Ypt1p) and it contained GTP-binding domains that are conserved among all known ras-like GTPases. Multicopy plasmids containing C. albicans YPT1 complemented the temperature-sensitive S. cerevisiae ypt1 (A136D) mutation. One chromosomal YPT1 allele in C. albicans CAI4 was readily disrupted by homologous gene targeting, but attempts to disrupt the second allele yielded no viable null mutants. Since this suggested that C. albicans YPT1 may be essential, a mutant ypt1 allele was constructed encoding the amino acid substitution analogous to the N121I substitution in a known trans-dominant inhibitor of S. cerevisiae Ypt1p. Next, a GAL1-regulated plasmid was used to express the mutant ypt1(N121I) allele in C. albicans CAI4. Ten of 11 transformants tested grew normally in glucose and poorly in galactose, and plasmid curing restored growth to wild-type levels. When these transformants were incubated in galactose, secretion of aspartyl proteinase (Sap) was inhibited and membrane-bound secretory vesicles accumulated intracellularly. These results imply that C. albicans YPT1 is required for growth and protein secretion, and they confirm the feasibility of using inducible dominant-negative alleles to define the functions of essential genes in C. albicans.


Non-VA primary care providers' perspectives on comanagement for rural veterans.
Many veterans utilize health care services within and outside of the Department of Veterans Affairs (VA). There are limited VA care coordination resources for non-VA primary care providers (PCPs), and the non-VA provider perspective on caring for veterans is underrepresented. The VA requires Patient Aligned Care Teams to coordinate care for veterans across health systems. To elicit perspectives of PCPs on caring for veterans who use both VA and non-VA health care. Qualitative data from semistructured telephone interviews were interpreted in the context of quantitative survey results. Participants were PCPs in a practice-based research network in 2011. 67 non-VA PCPs completed surveys, and descriptive statistics were performed. 21 semistructured telephone interviews were transcribed and underwent thematic analysis. Current communication with VA was viewed as poor, and many believed this led to poor patient outcomes. The veteran was identified as the main vehicle for information transfer between providers, which was viewed as undesirable. Non-VA providers felt they were interacting with VA as a system rather than with individual providers. VA system barriers hinder communication between providers, possibly resulting in fragmented care. Addressing these barriers will potentially improve patient safety and satisfaction.


Primary orthotopic glioma xenografts recapitulate infiltrative growth and isocitrate dehydrogenase I mutation.
Malignant gliomas constitute a heterogeneous group of highly infiltrative glial neoplasms with distinct clinical and molecular features. Primary orthotopic xenografts recapitulate the histopathological and molecular features of malignant glioma subtypes in preclinical animal models. To model WHO grades III and IV malignant gliomas in transplantation assays, human tumor cells are xenografted into an orthotopic site, the brain, of immunocompromised mice. In contrast to secondary xenografts that utilize cultured tumor cells, human glioma cells are dissociated from resected specimens and transplanted without prior passage in tissue culture to generate primary xenografts. The procedure in this report details tumor sample preparation, intracranial transplantation into immunocompromised mice, monitoring for tumor engraftment and tumor harvesting for subsequent passage into recipient animals or analysis. Tumor cell preparation requires 2 hr and surgical procedure requires 20 min/animal.


Fas-activated serine/threonine phosphoprotein (FAST) is a regulator of alternative splicing.
Fas-activated serine/threonine phosphoprotein (FAST) is a survival protein that is tethered to the outer mitochondrial membrane. In cells subjected to environmental stress, FAST moves to stress granules, where it interacts with TIA1 to modulate the process of stress-induced translational silencing. Both FAST and TIA1 are also found in the nucleus, where TIA1 promotes the inclusion of exons flanked by weak splice recognition sites such as exon IIIb of the fibroblast growth factor receptor 2 (FGFR2) mRNA. Two-hybrid interaction screens and biochemical analysis reveal that FAST binds to several alternative and constitutive splicing regulators, suggesting that FAST might participate in this process. The finding that FAST is concentrated at nuclear speckles also supports this contention. We show that FAST, like TIA1, promotes the inclusion of exon IIIb of the FGFR2 mRNA. Both FAST and TIA1 target a U-rich intronic sequence (IAS1) adjacent the 5' splice site of exon IIIb. However, unlike TIA1, FAST does not bind to the IAS1 sequence. Surprisingly, knockdown experiments reveal that FAST and TIA1 act independently of one another to promote the inclusion of exon IIIb. Mutational analysis reveals that FAST-mediated alternative splicing is separable from the survival effects of FAST. Our data reveal that nuclear FAST can regulate the splicing of FGFR2 transcripts.


The neurotoxic effects of estrogen on ischemic stroke in older female rats is associated with age-dependent loss of insulin-like growth factor-1.
Hormone therapy to postmenopausal females increases the risk and severity of ischemic stroke. Our previous work using an animal model of menopause (reproductive senescence) shows that middle cerebral artery occlusion (MCAo) causes a larger cortical-striatal infarct in this older acyclic group compared with younger females. Moreover, although estrogen treatment is neuroprotective in younger females, estrogen paradoxically increases infarct volume in acyclic females. We hypothesized that the neurotoxic effects of estrogen in older females occurs because of decreased availability of IGF-1, a neuroprotectant that decreases with advancing age and is downregulated by estrogen treatment. Our data show that plasma IGF-1 levels are significantly reduced in reproductive senescent females and further reduced by estrogen at all ages. The neuroprotective effect of estrogen on MCAo-induced cortical infarct volume in mature adult female is reversed by intracerebroventricular injections of IGF-1 receptor antagonist JB-1. Similarly, estrogens neurotoxic effects on cortical infarct volume in senescent females is attenuated by concurrent IGF-1 treatment, and reversed when IGF-1 is infused 4 h after the onset of ischemia (delayed IGF-1 treatment). Delayed IGF-1/estrogen treatment also suppressed ischemia-induced ERK1 phosphorylation, reduced protein oxidation, and stimulated an early increase in prostaglandin E(2) at the infarct site. IGF-1 treatment was only protective in senescent females that received estrogen, indicating that the neuroprotective actions of this peptide require interaction with the steroid hormone receptor. These data support the hypothesis that stroke severity in older females is associated with decreased IGF-1 and further indicate that short-term postischemic IGF-1 therapy may be beneficial for stroke.


Lower-than-expected CH4 emissions from rice paddies with rising CO2 concentrations.
Elevated atmospheric CO2 (eCO2 ) generally increases carbon input in rice paddy soils and stimulates the growth of methane-producing microorganisms. Therefore, eCO2 is widely expected to increase methane (CH4 ) emissions from rice agriculture, a major source of anthropogenic CH4 . Agricultural practices strongly affect CH4 emissions from rice paddies as well, but whether these practices modulate effects of eCO2 is unclear. Here we show, by combining a series of experiments and meta-analyses, that whereas eCO2 strongly increased CH4 emissions from paddies without straw incorporation, it tended to reduce CH4 emissions from paddy soils with straw incorporation. Our experiments also identified the microbial processes underlying these results: eCO2 increased methane-consuming microorganisms more strongly in soils with straw incorporation than in soils without straw, with the opposite pattern for methane-producing microorganisms. Accounting for the interaction between CO2 and straw management, we estimate that eCO2 increases global CH4 emissions from rice paddies by 3.7%, an order of magnitude lower than previous estimates. Our results suggest that the effect of eCO2 on CH4 emissions from rice paddies is smaller than previously thought and underline the need for judicious agricultural management to curb future CH4 emissions.


The EuroChimerism concept for a standardized approach to chimerism analysis after allogeneic stem cell transplantation.
Hematopoietic stem cell transplantation is becoming an increasingly important approach to treatment of different malignant and non-malignant disorders. There is thus growing demand for diagnostic assays permitting the surveillance of donor/recipient chimerism posttransplant. Current techniques are heterogeneous, rendering uniform evaluation and comparison of diagnostic results between centers difficult. Leading laboratories from 10 European countries have therefore performed a collaborative study supported by a European grant, the EuroChimerism Concerted Action, with the aim to develop a standardized diagnostic methodology for the detection and monitoring of chimerism in patients undergoing allogeneic stem cell transplantation. Following extensive analysis of a large set of microsatellite/short tandem repeat (STR) loci, the EuroChimerism (EUC) panel comprising 13 STR markers was established with the aim to optimally meet the specific requirements of quantitative chimerism analysis. Based on highly stringent selection criteria, the EUC panel provides multiple informative markers in any transplant setting. The standardized STR-PCR tests permit detection of donor- or recipient-derived cells at a sensitivity ranging between 0.8 and 1.6%. Moreover, the EUC assay facilitates accurate and reproducible quantification of donor and recipient hematopoietic cells. Wide use of the European-harmonized protocol for chimerism analysis presented will provide a basis for optimal diagnostic support and timely treatment decisions.


Prebiotic treatment of experimental colitis with germinated barley foodstuff: a comparison with probiotic or antibiotic treatment.
There is increasing evidence that intestinal microflora play an important role in the pathogenesis of ulcerative colitis. Therefore, modification of the microflora by prebiotics, probiotics, and antibiotics may be a rational approach for controlling intestinal inflammation. Germinated barley food-stuff (GBF) is an insoluble mixture of glutamine-rich protein and hemicellulose-rich dietary fiber. GBF is utilized efficiently by Bifidobacterium, Lactobacillus, and Eubacterium and converted by them into lactate, acetate, and butyrate. These bacterial organic acids preserve a favorable intestinal condition. We have previously shown that GBF has attenuated intestinal inflammation in patients with ulcerative colitis and experimental colitis models through prebiotic actions. The aim of this study was to compare the effect of GBF with that of probiotics and antibiotics in an experimental colitis model. Colitis was induced by feeding male SD rats with a diet containing 3.0-3.5% dextran sodium sulfate (DSS). The therapeutic effect of oral administration of a prebiotic (GBF), probiotics (mixture of Lactobacillus and Clostridium butyricum), antibiotics (vancomycin, metronidazole), and the vehicle was determined by assessing clinical and pathological scores on day 6 after initiation of colitis. Butyrate concentrations in the cecal content were also determined. GBF treatment significantly reduced colonic inflammation as assessed by clinical scores with an increase in cecal butyrate levels. Probiotic treatment with a mixture of Lactobacillus and Clostridium butyricum did not show such an effect. Both antibiotic treatments significantly attenuated clinical and pathological scores. However, in contrast to GBF, this treatment led to a significant decrease in cecal butyrate levels. These data suggest that modification of the intestinal microflora by prebiotics, including GBF, may serve as a useful adjunct in the treatment of ulcerative colitis as well as antibiotic treatment.


How the Neurotoxin β-N-Methylamino-l-Alanine Accumulates in Bivalves: Distribution of the Different Accumulation Fractions among Organs.
The environmental neurotoxin β-methylamino-l-alanine (BMAA) may represent a risk for human health. BMAA accumulates in freshwater and marine organisms consumed by humans. However, few data are available about the kinetics of BMAA accumulation and detoxification in exposed organisms, as well as the organ distribution and the fractions in which BMAA is present in tissues (free, soluble bound or precipitated bound cellular fractions). Here, we exposed the bivalve mussel Dreissena polymorpha to 7.5 µg of dissolved BMAA/mussel/3 days for 21 days, followed by 21 days of depuration in clear water. At 1, 3, 8, 14 and 21 days of exposure and depuration, the hemolymph and organs (digestive gland, the gills, the mantle, the gonad and muscles/foot) were sampled. Total BMAA as well as free BMAA, soluble bound and precipitated bound BMAA were quantified by tandem mass spectrometry. Free and soluble bound BMAA spread throughout all tissues from the first day of exposure to the last day of depuration, without a specific target organ. However, precipitated bound BMAA was detected only in muscles and foot from the last day of exposure to day 8 of depuration, at a lower concentration compared to free and soluble bound BMAA. In soft tissues (digestive gland, gonad, gills, mantle and muscles/foot), BMAA mostly accumulated as a free molecule and in the soluble bound fraction, with variations occurring between the two fractions among tissues and over time. The results suggest that the assessment of bivalve contamination by BMAA may require the quantification of total BMAA in whole individuals when possible.


Black and Garlic Mustard Plants Are Highly Suitable for the Development of Two Native Pierid Butterflies.
In multivoltine insects that oviposit and develop on short-lived plants, different herbivore generations across a growing season often exploit different plant species. Here, we compare the development time, pupal mass, and survival of two closely related oligophagous herbivore species on two species of brassicaceous plants that grow in different habitats and which exhibit little overlap in temporal growth phenology. In central Europe, the green-veined white butterfly, Pieris napi L., is bivoltine, whereas the small cabbage white butterfly, Pieris rapae L., has two to three generations a year. Moreover, P. napi is primarily found in moist, open (e.g., meadow), and forest habitats, whereas P. rapae prefers drier, open habitats. Both butterflies were reared on Garlic mustard (Alliaria petiolata), which is shade-tolerant and grows early in spring in forest undergrowth, and Black mustard (Brassica nigra), which prefers open disturbed habitats and is most common in summer. Both host plant species differ in other traits such as secondary chemistry. We hypothesized that, owing to habitat preference, P. napi would develop equally well on both plants but that P. rapae would perform better on B. nigra. The results provide partial support for this hypothesis, as both herbivores performed equally well on A. petiolata and B. nigra. However, there were differences in these parameters that were species-specific: on both plants P. rapae developed faster and had larger pupae than P. napi. Our results show that specialized herbivores can exploit different species of related plants that grow at different times of the season, enabling them to have multiple generations.


Hyperuricemia treatment in acute heart failure patients does not improve their long-term prognosis: A propensity score matched analysis from the AHEAD registry.
Hyperuricemia is associated with a poorer prognosis in heart failure (HF) patients. Benefits of hyperuricemia treatment with allopurinol have not yet been confirmed in clinical practice. The aim of our work was to assess the benefit of allopurinol treatment in a large cohort of HF patients. The prospective acute heart failure registry (AHEAD) was used to select 3160 hospitalized patients with a known level of uric acid (UA) who were discharged in a stable condition. Hyperuricemia was defined as UA ≥500 μmoL/L and/or allopurinol treatment at admission. The patients were classified into three groups: without hyperuricemia, with treated hyperuricemia, and with untreated hyperuricemia at discharge. Two- and five-year all-cause mortality were defined as endpoints. Patients without hyperuricemia, unlike those with hyperuricemia, had a higher left ventricular ejection fraction, a better renal function, and higher hemoglobin levels, had less frequently diabetes mellitus and atrial fibrillation, and showed better tolerance to treatment with angiotensin-converting enzyme inhibitors/angiotensin receptor blockers and/or beta-blockers. In a primary analysis, the patients without hyperuricemia had the highest survival rate. After using the propensity score to set up comparable groups, the patients without hyperuricemia had a similar 5-year survival rate as those with untreated hyperuricemia (42.0% vs 39.7%, P = 0.362) whereas those with treated hyperuricemia had a poorer prognosis (32.4% survival rate, P = 0.006 vs non-hyperuricemia group and P = 0.073 vs untreated group). Hyperuricemia was associated with an unfavorable cardiovascular risk profile in HF patients. Treatment with low doses of allopurinol did not improve the prognosis of HF patients.


Bayesian validation of grammar productions for the language of thought.
Probabilistic proposals of Language of Thoughts (LoTs) can explain learning across different domains as statistical inference over a compositionally structured hypothesis space. While frameworks may differ on how a LoT may be implemented computationally, they all share the property that they are built from a set of atomic symbols and rules by which these symbols can be combined. In this work we propose an extra validation step for the set of atomic productions defined by the experimenter. It starts by expanding the defined LoT grammar for the cognitive domain with a broader set of arbitrary productions and then uses Bayesian inference to prune the productions from the experimental data. The result allows the researcher to validate that the resulting grammar still matches the intuitive grammar chosen for the domain. We then test this method in the language of geometry, a specific LoT model for geometrical sequence learning. Finally, despite the fact of the geometrical LoT not being a universal (i.e. Turing-complete) language, we show an empirical relation between a sequence's probability and its complexity consistent with the theoretical relationship for universal languages described by Levin's Coding Theorem.


Highly efficient genome editing of human hematopoietic stem cells via a nano-silicon-blade delivery approach.
Recently, the clustered regularly interspaced short palindromic repeats (CRISPR)-Cas9 bacterial immunity system has opened a promising avenue to treat genetic diseases that affect the human hematopoietic stem cells (HSCs). Therefore, finding a highly efficient delivery method capable of modifying the genome in the hard-to-transfect HSCs, combined with the advanced CRISPR-Cas9 system, may meet the challenges for dissecting the hematologic disease mechanisms and facilitate future clinical applications. Here, we developed an effective HSC-specified delivery microfluidic chip to disrupt the cell membrane transiently by inducing rapid mechanical deformation that allowed the delivery of biomaterials into the cytoplasm from the surrounding matrix. Compared with the previous designs, the new nano-silicon-blade structure was specifically optimized for HSCs. Using the silicon substrate, the sharpness and rigidity of the nano-blade constriction was largely enhanced to improve the biomaterials delivery efficiency. We achieved highly efficient delivery results by transporting various macro-molecules into the HSCs. Moreover, the treated HSCs possess high viability and maintain inherent pluripotency after the delivery via the Nano-Blade Chip (NB-Chip). Subsequently, we disrupted the p42 isoform in C/EBPα on the NB-Chip and induced HSCs into a myeloid proliferation behavior. In conclusion, the NB-Chip provides a harmless, rapid and high-throughput gene editing approach for the HSC study and therapeutics.


Stability of normal personality traits after traumatic brain injury.
To test the hypothesis that changes in personality traits are evident after traumatic brain injury (TBI) using current models of normal adult personality variation. Comparison of inception cohort and control group at two measurement occasions. A large urban academic medical center. Retrospective personality assessments were obtained from significant others of 21 TBI patients within 30 days of injury and at 6-month follow-up and from a control group of significant others of 25 persons without neurological history twice over a 6-month interval. Five scales-Neuroticism, Extraversion, Openness to Experience, Agreeableness, and Conscientiousness-from the revised NEO Personality Inventory (NEO PI-R), Form R, and an observer rating scale for retrospective estimates of change (REC). Significant score changes were found for only one of the five trait domains in the patient sample; controls showed minimal changes overall. Patients' Extraversion scores declined to average levels at 6-month follow-up, diminishing premorbid differences between patients and controls on this dimension. Subjective change estimates made by raters after follow-up reflected perceptions of increased neuroticism in patients that were inconsistent with the serial NEO PI-R data the raters provided. The absence of systematic changes in personality trait scores among the patients cautions against presuming that such changes account for the behavior of TBI patients.


Exact analytic flux distributions for two-dimensional solar concentrators.
A new approach for representing and evaluating the flux density distribution on the absorbers of two-dimensional imaging solar concentrators is presented. The formalism accommodates any realistic solar radiance and concentrator optical error distribution. The solutions obviate the need for raytracing, and are physically transparent. Examples illustrating the method's versatility are presented for parabolic trough mirrors with both planar and tubular absorbers, Fresnel reflectors with tubular absorbers, and V-trough mirrors with planar absorbers.


Regulatory T cell enrichment by IFN-γ conditioning.
IFN-γ was originally characterized as a proinflammatory cytokine with T helper type 1 inducing activity, but it is now clear that it also has important immunoregulatory functions. Regulatory T cells play an important role in models of autoimmunity, GVHD, and transplantation, and offer potential as a cellular therapy. In rodent models, in vivo-generated CD25(+)CD4(+) T cells can prevent allograft rejection, but therapeutic exploitation of Treg will more likely depend on protocols that allow the generation or selection of Treg ex vivo. The experiments described in this chapter will show that alloantigen-reactive Treg can be generated/expanded ex vivo using IFN-γ, a cytokine more usually associated with allograft rejection. Although IFN-γ production has hitherto been generally regarded as nonpermissive for allograft survival, we believe this paradoxical "good-bad" role for IFN-γ may reflect an important physiological negative feedback loop.


Frequency and characteristic features of portal hypertensive gastropathy in patients with viral cirrhosis.
To determine the frequency and specific characteristic features of portal hypertensive gastropathy (PHG) in cirrhosis due to viral etiology. Cross-sectional descriptive study. The Aga Khan University Hospital, Karachi, from June 2006 till June 2008. Patients with hepatitis B and C cirrhosis were included who underwent screening esophago-gastroduodenoscopy (EGD) for varices. Baveno III consensus statement was used for diagnosing PHG on endoscopy and divided them into two subgroups i.e. mild and severe. Data related to platelet/spleen ratio, MELD score and Child Turcotte Pugh (CTP) score indicating severity of cirrhosis were recorded in all patients. Findings were compared by using independent sample t-test. Out of 360 patients who underwent screening EGD, 226 (62.8%) were males. Two hundred and eighty one (78%) had hepatitis C while 79 (22%) suffered from hepatitis B related cirrhosis. Three hundred patients (83.3%) had PHG while 71 (24%) had severe PHG. Higher proportion of esophageal varices (89.7%) was present among those who had PHG (p < 0.001). On univariate analysis lower platelet counts (117±55 vs. 167±90; p < 0.001), increased spleen size (14.1±2.9 cm vs. 12±2.4cm; p < 0.001) were found in PHG patients as compared to those without it. Similarly, lower platelet/spleen ratio was noted in patients with severe PHG (916±400 vs. 1477±899; p < 0.001). Furthermore, on multivariate analysis CTP score > 8 MELD score > 12 and platelets/spleen ratio < 900 were significantly associated factors with severe PHG. Frequency of PHG was 83% while severe PHG was seen in 24% cases of viral hepatic cirrhosis. MELD score > 12, CTP score ³ 8 and platelets/spleen ratio < 900 were significant factors of severe PHG.


Maturation of granule cell dendrites after mossy fiber arrival in hippocampal field CA3.
Most granule neurons in the rat dentate gyrus are born over the course of the first 2 postnatal weeks. The resulting heterogeneity has made it difficult to define the relationship between dendritic and axonal maturation and to delineate a time course for the morphological development of the oldest granule neurons. By depositing crystals of the fluorescent label Dil in hippocampal field CA3, we retrogradely labeled granule neurons in fixed tissue slices from rats aged 2-9 days. The results showed that all labeled granule cells, regardless of the age of the animal, exhibited apical dendrites. On day 2, every labeled neuron had rudimentary apical dendrites, and a few dendrites on each cell displayed immature features such as growth cones, varicosities, and filopodia. Some cells displayed basal dendrites. By day 4, the most mature granule neurons had longer and more numerous apical branches, as well as various immature features. Most had basal dendrites. On days 5 and 6, the immature features and the basal dendrites had begun to regress on the oldest cells, and varying numbers of spines were present. On day 7, the first few adult-like neurons were seen: immature features and basal dendrites had disappeared, all dendrites reached the top of the molecular layer, and the entire dendritic tree was covered with spines. These data show that dendritic outgrowth occurs before, or concurrent with, axon arrival in the CA3 target region, and that adult-like granule neurons are present by the end of the first week.


PARP1 inhibitor olaparib (Lynparza) exerts synthetic lethal effect against ligase 4-deficient melanomas.
Cancer including melanoma may be ''addicted" to double strand break (DSB) repair and targeting this process could sensitize them to the lethal effect of DNA damage. PARP1 exerts an important impact on DSB repair as it binds to both single- and double- strand breaks. PARP1 inhibitors might be highly effective drugs triggering synthetic lethality in patients whose tumors have germline or somatic defects in DNA repair genes. We hypothesized that PARP1-dependent synthetic lethality could be induced in melanoma cells displaying downregulation of DSB repair genes. We observed that PARP1 inhibitor olaparib sensitized melanomas with reduced expression of DNA ligase 4 (LIG4) to an alkylatimg agent dacarbazine (DTIC) treatment in vitro, while normal melanocytes remained intact. PARP1 inhibition caused accumulation of DSBs, which was associated with apoptosis in LIG4 deficient melanoma cells. Our hypothesis that olaparib is synthetic lethal with LIG4 deficiency in melanoma cells was supported by selective anti-tumor effects of olaparib used either alone or in combination with dacarbazine (DTIC) in LIG4 deficient, but not LIG4 proficient cells. In addition, olaparib combined with DTIC inhibited the growth of LIG4 deficient human melanoma xenografts. This work for the first time demonstrates the effectiveness of a combination of PARP1 inhibitor olaparib and alkylating agent DTIC for treating LIG4 deficient melanomas. In addition, analysis of the TCGA and transcriptome microarray databases revealed numerous individual melanoma samples potentially displaying specific defects in DSB repair pathways, which may predispose them to synthetic lethality triggered by PARP1 inhibitor combined with a cytotoxic drug.


The role of antennae in removing entomopathogenic fungi from cuticle of the termite, Coptotermes formosanus.
Our previous research has shown that the termite, Coptotermes formosanus Shiraki (Isoptera: Rhinotermitidae), protects itself from entomopathogenic fungi by mutual grooming behavior. The termite removes and discards foreign organisms, such as fungal conidia, from the body surface of its nestmates by mutual grooming behavior. The role of the antennae in detecting the condia was examind here. Three entomopathogenic fungi were used, Beauveria brongniartii 782 (Saccardo) (Hypocreales), Paecilomyces fumosoroseus K3 (Wize) (Hyphomycetes), and Metarhizium anisopliae 455 Sorokin (Hyphomycetes). Termites with antennae removed conidia more efficiently than termites without antennae. There were differences between termites with and without antennae in selection of sites to be groomed on nestmates, in the length of grooming and in occurrence of grooming. Electroantennogram (EAG) responses were recorded from termite antennae and the waveforms were rather specific to the kinds of fungi used as odor sources. Termites were able to distinguish between the tested fungi in feeding tests. These results show that the antennae play important roles in the mutual grooming behavior of the termite.


The influence of chloride deicers on mineral nutrition and the health status of roadside trees in the city of Kielce, Poland.
Application of chemical road deicers has a negative impact on roadside vegetation. Every year, the trees in cities suffer from direct and indirect effects of salt application for winter road maintenance. To elucidate this problem in the city of Kielce, the chemistry of snow, soil, tree bark, and leaf samples has been investigated together with an assessment of the health status of the trees. Twelve investigation sites were selected along the roads with different traffic intensity. Snow samples were collected twice during the winter and analyzed for pH, EC, Na(+), Ca(2+), Mg(2+), and Cl(-). In soil (collected from two depth intervals), tree bark, and leaf samples, the concentrations of B, Ca, Fe, K, Mg, Mn, N, Na, P, S, and Zn were determined. The contents of total organic carbon (TOC) in soils, as well as the pH of soil and tree bark samples were also measured. Negative symptoms revealed by roadside trees included the loss of assimilative apparatus and decreased vitality. The results of chemical analyses indicated that the snowmelt might be a substantial source of chloride ions and alkalizing substances that influenced higher pH of soils. The soil samples displayed elevated concentrations of S and Zn and lower than typical for soil contents of B, Mg, and TOC. The pH of alkaline soils favored greater bioavailability of B and reduced bioavailability of Na and Zn by the trees examined.


Sex-biased dispersal, kin selection and the evolution of sexual conflict.
There is growing interest in resolving the curious disconnect between the fields of kin selection and sexual selection. Rankin's (2011, J. Evol. Biol. 24, 71-81) theoretical study of the impact of kin selection on the evolution of sexual conflict in viscous populations has been particularly valuable in stimulating empirical research in this area. An important goal of that study was to understand the impact of sex-specific rates of dispersal upon the coevolution of male-harm and female-resistance behaviours. But the fitness functions derived in Rankin's study do not flow from his model's assumptions and, in particular, are not consistent with sex-biased dispersal. Here, we develop new fitness functions that do logically flow from the model's assumptions, to determine the impact of sex-specific patterns of dispersal on the evolution of sexual conflict. Although Rankin's study suggested that increasing male dispersal always promotes the evolution of male harm and that increasing female dispersal always inhibits the evolution of male harm, we find that the opposite can also be true, depending upon parameter values.


Malaria transmission and morbidity.
Stable malaria endemicity is maintained over a wide range of transmission intensities in sub-Saharan Africa. This paper considers variations in the clinical manifestations and their consequences with differences in transmission intensity. Epidemiological approaches to malarial disease have concentrated on two clinical syndromes, severe malarial anaemia and cerebral malaria. Within an area the mean age of children with severe malarial anaemia is always lower than that of those with cerebral malaria. In areas of higher malaria transmission children, on average, encounter malaria at a younger age and the mean age of clinical cases is lower. Malarial anaemia tends therefore to be relatively more important under high transmission settings and cerebral malaria tends to gain in importance under lower transmission settings. In a number of studies the total load of malaria morbidity, whether measured as none severe malaria in the community or as severe malaria admitted to hospital, is low under stable low transmission conditions but is at its highest under moderate intensities of transmission. Thereafter it reaches a plateau, or even falls, at the highest transmission intensities. It is not known whether the same is true for mortality in communities living under different transmission settings. Possible implications for changes in patterns of morbidity and mortality following interventions which lower malaria transmission are discussed. It is concluded that such interventions should play an important role in integrated malaria control programmes but that these should involve concomitant introduction of other interventions, in order to minimise the possible risks of a reduced effect as the immune response of the population re-equilibrates in the face of reduced challenge.


Fatal encephalitis caused by concomitant infection with tick-borne encephalitis virus and Borrelia burgdorferi.
We describe a 38-year-old farmer from the southwestern archipelago of Finland where both tick-borne encephalitis (TBE) virus and Borrelia burgdorferi are endemic. He presented with fever and headache, developed severe meningoencephalitis in 3 days, and, after 1 month, died without regaining consciousness. High titers of IgG and IgM antibodies to TBE virus were present in both serum and CSF. Serology for Borrelia was negative. Autopsy revealed necrotizing encephalitis and myelitis with involvement of the dorsal root ganglion. With use of polymerase chain reaction tests, segments of two separate genes of B. burgdorferi were amplified from the patient's CSF. This case demonstrates that the possibility of dual infection should be considered for patients residing in geographic areas where Ixodes ticks may carry both the TBE virus and B. burgdorferi. We believe that the most severe damage in this case was caused by TBE virus rather than by B. burgdorferi. Nevertheless, the coinfection might have contributed to the fatal outcome that has not been previously observed in Finnish patients with TBE.


Cerebral aneurysms causing visual symptoms: their features and surgical outcome.
Cerebral aneurysms causing visual symptoms before surgery are relatively rare. We have experience with 17 cases of such aneurysms and report their clinical features and surgical outcome. The locations of aneurysms presenting with visual dysfunction in our series are as follows: internal carotid (IC)-cavernous aneurysms in six of 29 total cases, 21%; IC-ophthalmic aneurysms in nine of 36 total cases, 25%; and anterior communicating artery (A com A) aneurysms in two of 217 total cases, 1%. The size of the aneurysms, the period between the onset of symptoms and surgical treatment, the pre- and post-operative visual function, and the surgical methods used to treat the aneurysm were analyzed. All the visually symptomatic cases featured large (15-24 mm) or giant (> 25 mm) aneurysms. Visual symptoms occurred before aneurysmal rupture in all cases but one. The type of visual field defect and the degree of reduced visual acuity were highly variable, without a typical clinical presentation. Five out of six IC-cavernous aneurysms were treated surgically with common carotid artery (CC) ligation or IC ligation with superficial temporal artery (STA) to middle cerebral artery (MCA) anastomosis. Three of these cases showed improvement of visual symptoms after surgery. Six out of nine IC-ophthalmic aneurysms were treated surgically (CC ligation or direct clipping), with four cases showing improvement of visual symptoms after surgery. One case of an A com A aneurysm featured a ruptured aneurysm that had physically penetrated the optic chiasm, while the other case was a giant unruptured aneurysm. The interval between the onset of symptoms and surgical treatment was the only factor identified which affected the clinical outcome of the aneurysms presenting with visual dysfunction. All cases that were determined to show improvement of visual function were treated surgically within 3 months of the onset of symptoms. Cerebral aneurysms presenting with visual dysfunction before surgery are most commonly large or giant, and unruptured. Recovery of visual function can most often be expected when surgical treatment is performed expeditiously, before the visual dysfunction becomes irreversible.


Parallel discovery of Alzheimer's therapeutics.
As the prevalence of Alzheimer's disease (AD) grows, so do the costs it imposes on society. Scientific, clinical, and financial interests have focused current drug discovery efforts largely on the single biological pathway that leads to amyloid deposition. This effort has resulted in slow progress and disappointing outcomes. Here, we describe a "portfolio approach" in which multiple distinct drug development projects are undertaken simultaneously. Although a greater upfront investment is required, the probability of at least one success should be higher with "multiple shots on goal," increasing the efficiency of this undertaking. However, our portfolio simulations show that the risk-adjusted return on investment of parallel discovery is insufficient to attract private-sector funding. Nevertheless, the future cost savings of an effective AD therapy to Medicare and Medicaid far exceed this investment, suggesting that government funding is both essential and financially beneficial.


A unified model for functional and statistical epistasis and its application in quantitative trait Loci analysis.
Interaction between genes, or epistasis, is found to be common and it is a key concept for understanding adaptation and evolution of natural populations, response to selection in breeding programs, and determination of complex disease. Currently, two independent classes of models are used to study epistasis. Statistical models focus on maintaining desired statistical properties for detection and estimation of genetic effects and for the decomposition of genetic variance using average effects of allele substitutions in populations as parameters. Functional models focus on the evolutionary consequences of the attributes of the genotype-phenotype map using natural effects of allele substitutions as parameters. Here we provide a new, general and unified model framework: the natural and orthogonal interactions (NOIA) model. NOIA implements tools for transforming genetic effects measured in one population to the ones of other populations (e.g., between two experimental designs for QTL) and parameters of statistical and functional epistasis into each other (thus enabling us to obtain functional estimates of QTL), as demonstrated numerically. We develop graphical interpretations of functional and statistical models as regressions of the genotypic values on the gene content, which illustrates the difference between the models--the constraint on the slope of the functional regression--and when the models are equivalent. Furthermore, we use our theoretical foundations to conceptually clarify functional and statistical epistasis, discuss the advantages of NOIA over previous theory, and stress the importance of linking functional and statistical models.


Systems Biology Reveals Cigarette Smoke-Induced Concentration-Dependent Direct and Indirect Mechanisms That Promote Monocyte-Endothelial Cell Adhesion.
Cigarette smoke (CS) affects the adhesion of monocytes to endothelial cells, a critical step in atherogenesis. Using an in vitro adhesion assay together with innovative computational systems biology approaches to analyze omics data, our study aimed at investigating CS-induced mechanisms by which monocyte-endothelial cell adhesion is promoted. Primary human coronary artery endothelial cells (HCAECs) were treated for 4 h with (1) conditioned media of human monocytic Mono Mac-6 (MM6) cells preincubated with low or high concentrations of aqueous CS extract (sbPBS) from reference cigarette 3R4F for 2 h (indirect treatment, I), (2) unconditioned media similarly prepared without MM6 cells (direct treatment, D), or (3) freshly generated sbPBS (fresh direct treatment, FD). sbPBS promoted MM6 cells-HCAECs adhesion following I and FD, but not D. In I, the effect was mediated at a low concentration through activation of vascular inflammation processes promoted in HCAECs by a paracrine effect of the soluble mediators secreted by sbPBS-treated MM6 cells. Tumor necrosis factor α (TNFα), a major inducer, was actually shed by unstable CS compound-activated TNFα-converting enzyme. In FD, the effect was triggered at a high concentration that also induced some toxicity. This effect was mediated through an yet unknown mechanism associated with a stress damage response promoted in HCAECs by unstable CS compounds present in freshly generated sbPBS, which had decayed in D unconditioned media. Aqueous CS extract directly and indirectly promotes monocytic cell-endothelial cell adhesion in vitro via distinct concentration-dependent mechanisms.


Medication-related fall incidents in an older, ambulant population: the B-PROOF study.
Medication use is a potentially modifiable risk factor for falling; psychotropic and cardiovascular drugs have been indicated as main drug groups that increase fall risk. However, evidence is mainly based on studies that recorded falls retrospectively and/or did not determine medication use at the time of the fall. Therefore, we investigated the associations indicated in the literature between medication use and falls, using prospectively recorded falls and medication use determined at the time of the fall. Data from the B-PROOF (B-vitamins for the prevention of osteoporotic fractures) study were used, concerning community-dwelling elderly aged ≥65 years. We included 2,407 participants with pharmacy dispensing records. During the 2- to 3-year follow-up, participants recorded falls using a fall calendar. Cox proportional hazard models were applied, adjusting for potential confounders including age, sex, health status variables and concomitant medication use. During follow-up, 1,147 participants experienced at least one fall. Users of anti-arrhythmic medication had an increased fall risk (hazard ratio [HR] 1.61; 95% confidence interval [CI] 1.12-2.32) compared with non-users. Similarly, non-selective beta-blocker use was associated with an increased fall risk (HR 1.41 [95% CI 1.12-1.78]), while statin use was associated with a lower risk (HR 0.81 [95% CI 0.71-0.94]). Benzodiazepine use (HR 1.32 [95% CI 1.02-1.71]), and antidepressant use (HR 1.40 [95% CI 1.07-1.82]) were associated with an increased fall risk. Use of other cardiovascular and psychotropic medication was not associated with fall risk. Our results strengthen the evidence for an increased fall risk in community-dwelling elderly during the use of anti-arrhythmics, non-selective beta-blockers, benzodiazepines, and antidepressant medication. Clinicians should prescribe these drugs cautiously and if possible choose safer alternatives for older patients.


Angiogenic role of LYVE-1-positive macrophages in adipose tissue.
Here we report the discovery of a characteristic dense vascular network (DVN) in the tip portion of epididymal adipose tissue in adult mice. The DVN is formed by angiogenesis rather than by vasculogenesis, and has functional blood circulation. This DVN and its subsequent branching may provide a new functional route for adipogenesis. The recruitment, infiltration, and accumulation of bone marrow-derived LYVE-1(+) macrophages in the tip region are crucial for the formation of the DVN. Matrix metalloproteinases (MMPs) and the VEGF-VEGFR2 system are responsible not only for the formation of the DVN, but also for the recruitment and infiltration of LYVE-1(+) macrophages into the epididymal adipose tissue tip region. SDF-1, but not the MCP-1-CCR2 system, is a critical factor in recruitment and ongoing retention of macrophages in this area. We also demonstrate that the tip region of epididymal adipose tissue is highly hypoxic, and thus provides a microenvironment conducive to the high expression and enhanced activities of VEGF, VEGFR2, MMPs, and SDF-1 in autocrine and paracrine manners, to create an ideal niche for the recruitment, retention, and angiogenic action of macrophages. These findings shed light on the complex interplay between macrophage infiltration, angiogenesis, and adipogenesis in the tip region of adult epididymal adipose tissue, and provide novel insight into the regulation of alternative outgrowth of adipose tissue.


Deciphering conjugative plasmid permissiveness in wastewater microbiomes.
Wastewater treatment plants (WWTPs) are designed to robustly treat polluted water. They are characterized by ceaseless flows of organic, chemical and microbial matter, followed by treatment steps before environmental release. WWTPs are hotspots of horizontal gene transfer between bacteria via conjugative plasmids, leading to dissemination of potentially hazardous genetic material such as antimicrobial resistance genes (AMRGs). While current focus is on the threat of AMRGs spreading and their environmental maintenance, conjugative plasmid transfer dynamics within and between bacterial communities still remains largely uncharted. Furthermore, current in vitro methods used to assess conjugation in complex microbiomes do not include in situ behaviours of recipient cells, resulting in partial understanding of transfers. We investigated the in vitro conjugation capacities of WWTP microbiomes from inlet sewage and outlet treated water using the broad-host range IncP-1 conjugative plasmid, pKJK5. A thorough molecular approach coupling metagenomes to 16S rRNA DNA/cDNA amplicon sequencing was established to characterize microbiomes using the ecological concept of functional response groups. A broad diversity of recipient bacterial phyla for the plasmid was observed, especially in WWTP outlets. We also identified permissive bacteria potentially able to cross WWTPs and engage in conjugation before and after water treatment. Bacterial activity and lifestyle seem to influence conjugation extent, as treated water copiotrophs were the most represented strategist amongst transconjugants. Correlation analysis highlighted possible plasmid transmission routes into communities between the sewage to the environment, with identification of keystone members (e.g., Arcobacter) potentially involved in cross-border exchanges between distant Gram-positive and Gram-negative phyla.


Cost effectiveness of a 92-gene assay for the diagnosis of metastatic cancer.
To estimate the clinical and economic trade-offs involved in using a molecular assay (92-gene assay, CancerTYPE ID) to aid in identifying the primary site of difficult-to-diagnose metastatic cancers and to explore whether the 92-gene assay can be used to standardize the diagnostic process and costs for clinicians, patients, and payers. Four decision-analytic models were developed to project the lifetime clinical and economic impact of incorporating the 92-gene assay compared with standard care alone. For each model, total and incremental costs, life-years, quality-adjusted life-years (QALYs), incremental cost-effectiveness ratios (ICERs), and the proportion of patients treated correctly versus incorrectly were projected from the payer perspective. Model inputs were based on published literature, analyses of SEER (Surveillance Epidemiology and End RESULTS) data, publicly available data, and interviews with clinical experts. In all four models, the 92-gene assay increased the proportion of patients treated correctly, decreased the proportion of patients treated with empiric therapy, and increased quality-adjusted survival. In the primary model, the ICER was $50,273/QALY; thus, the 92-gene assay is therefore cost effective when considering a societal willingness-to-pay threshold of $100,000/QALY. These findings were robust across sensitivity analyses. Use of the 92-gene assay for diagnosing metastatic tumors of uncertain origin is associated with reduced misdiagnoses, increased survival, and improved quality of life. Incorporating the assay into current practice is a cost-effective approach to standardizing diagnostic methods while improving patient care. Limitations of this analysis are the lack of data availability and resulting modeling simplifications, although sensitivity analyses showed these to not be key drivers of results.


Maintenance DFMO Increases Survival in High Risk Neuroblastoma.
High risk neuroblastoma (HRNB) accounts for 15% of all pediatric cancer deaths. Despite aggressive therapy approximately half of patients will relapse, typically with only transient responses to second-line therapy. This study evaluated the ornithine decarboxylase inhibitor difluoromethylornithine (DFMO) as maintenance therapy to prevent relapse following completion of standard therapy (Stratum 1) or after salvage therapy for relapsed/refractory disease (Stratum 2). This Phase II single agent, single arm multicenter study enrolled from June 2012 to February 2016. Subjects received 2 years of oral DFMO (750 ± 250 mg/m2 twice daily). Event free survival (EFS) and overall survival (OS) were determined on an intention-to-treat (ITT) basis. 101 subjects enrolled on Stratum 1 and 100 were eligible for ITT analysis; two-year EFS was 84% (±4%) and OS 97% (±2%). 39 subjects enrolled on Stratum 2, with a two-year EFS of 54% (±8%) and OS 84% (±6%). DFMO was well tolerated. The median survival time is not yet defined for either stratum. DFMO maintenance therapy for HRNB in remission is safe and associated with high EFS and OS. Targeting ODC represents a novel therapeutic mechanism that may provide a new strategy for preventing relapse in children with HRNB.


Haplogroup effects and recombination of mitochondrial DNA: novel clues from the analysis of Leber hereditary optic neuropathy pedigrees.
The mitochondrial DNA (mtDNA) of 87 index cases with Leber hereditary optic neuropathy (LHON) sequentially diagnosed in Italy, including an extremely large Brazilian family of Italian maternal ancestry, was evaluated in detail. Only seven pairs and three triplets of identical haplotypes were observed, attesting that the large majority of the LHON mutations were due to independent mutational events. Assignment of the mutational events into haplogroups confirmed that J1 and J2 play a role in LHON expression but narrowed the association to the subclades J1c and J2b, thus suggesting that two specific combinations of amino acid changes in the cytochrome b are the cause of the mtDNA background effect and that this may occur at the level of the supercomplex formed by respiratory-chain complexes I and III. The families with identical haplotypes were genealogically reinvestigated, which led to the reconnection into extended pedigrees of three pairs of families, including the Brazilian family with its Italian counterpart. The sequencing of entire mtDNA samples from the reconnected families confirmed the genealogical reconstruction but showed that the Brazilian family was heteroplasmic at two control-region positions. The survey of the two sites in 12 of the Brazilian subjects revealed triplasmy in most cases, but there was no evidence of the tetraplasmy that would be expected in the case of mtDNA recombination.


Continuous flash suppression and monocular pattern masking impact subjective awareness similarly.
Peters and Lau (eLife, 4, e09651, 2015) found that when criterion bias is controlled for, there is no evidence for unconscious visual perception in normal observers, in the sense that they cannot directly discriminate a target above chance without knowing it. One criticism of that study is that the visual suppression method used, forward and backward masking (FBM), may be too blunt in the way it interferes with visual processing to allow for unconscious forced-choice discrimination. To investigate this question, we compared FBM directly to continuous flash suppression (CFS) in a two-interval forced-choice task. Although CFS is popular, and may be thought of as a more powerful visual suppression technique, we found no difference in the degree of perceptual impairment between the two suppression types. To the extent that CFS impairs perception, both objective discrimination and subjective awareness are impaired to similar degrees under FBM. This pattern was consistently observed across three experiments in which various experimental parameters were varied. These findings provide evidence for an ongoing debate about unconscious perception: normal observers cannot perform forced-choice discrimination tasks unconsciously.


PDM-iRZ-QPSK vs. PS-QPSK at 100 Gbit/s over dispersion-managed links.
We compare by simulation the performance of 100 Gbit/s PDM-iRZ-QPSK and PS-QPSK transmission both in homogeneous and hybrid QPSK/OOK DM links. We detail the reasons of the overall performance investigating each nonlinear effect (SPM, XPM and XPolM) individually. Moreover, we compare the accuracy of the noise loading method with the more realistic use of noisy in-line amplifiers. Results shows that i) PDM-iRZ-QPSK and PS-QPSK have same reach in both homogeneous and hybrid setups, ii) correct simulations must include distributed ASE.


Kinematic and EMG activities during front and back squat variations in maximum loads.
The aim of this study was to compare the musculature activity and kinematics of knee and hip joints during front and back squat with maximal loading. Two-dimensional kinematical data were collected and electromyographic activities of vastus lateralis, vastus medialis, rectus femoris, semitendinosus, biceps femoris, gluteus maximus and erector spinae were measured while participants (n = 12, 21.2 ± 1.9 years old) were completing front and back squat exercises with maximum loading. Paired sample t-test was used for comparisons between two techniques. Results showed that the electromyographic activity of vastus medialis was found to be greater in the front squat compared to the back squat during the ascending phase (P < 0.05, d = 0.62; 95% CI, -15.0/-4.17) and the whole manoeuvre (P < 0.05, d = 0.41; 95% CI, -12.8/-0.43), while semitendinosus (P < 0.05, d = -0.79; 95% CI, 0.62/20.59) electromyographic activity was greater in the back squat during the ascending phase. Compared to the front squat version, back squat exhibited significantly greater trunk lean, with no differences occurring in the knee joint kinematics throughout the movement. Results may suggest that the front squat may be preferred to the back squat for knee extensor development and for preventing possible lumbar injuries during maximum loading.


Proteogenomics Uncovers a Vast Repertoire of Shared Tumor-Specific Antigens in Ovarian Cancer.
High-grade serous ovarian cancer (HGSC), the principal cause of death from gynecologic malignancies in the world, has not significantly benefited from advances in cancer immunotherapy. Although HGSC infiltration by lymphocytes correlates with superior survival, the nature of antigens that can elicit anti-HGSC immune responses is unknown. The goal of this study was to establish the global landscape of HGSC tumor-specific antigens (TSA) using a mass spectrometry pipeline that interrogated all reading frames of all genomic regions. In 23 HGSC tumors, we identified 103 TSAs. Classic TSA discovery approaches focusing only on mutated exonic sequences would have uncovered only three of these TSAs. Other mutated TSAs resulted from out-of-frame exonic translation (n = 2) or from noncoding sequences (n = 7). One group of TSAs (n = 91) derived from aberrantly expressed unmutated genomic sequences, which were not expressed in normal tissues. These aberrantly expressed TSAs (aeTSA) originated primarily from nonexonic sequences, in particular intronic (29%) and intergenic (22%) sequences. Their expression was regulated at the transcriptional level by variations in gene copy number and DNA methylation. Although mutated TSAs were unique to individual tumors, aeTSAs were shared by a large proportion of HGSCs. Taking into account the frequency of aeTSA expression and HLA allele frequencies, we calculated that, in Caucasians, the median number of aeTSAs per tumor would be five. We conclude that, in view of their number and the fact that they are shared by many tumors, aeTSAs may be the most attractive targets for HGSC immunotherapy.


Levels and sources of stress in medical students.
Levels of stress, as measured by the general health questionnaire, were assessed in 318 medical students in their fourth year at three British universities. Mean scores were higher than those in other groups within the general population, and the estimated prevalence of emotional disturbance was 31.2%, a proportion similar to that reported in medical students in the United States. There were no differences in prevalence or in mean scores of stress between the sexes. Twelve (4%) students reported high intake of alcohol, and almost half of the students had increased their intake in the past two years. The four categories most commonly cited in answers to an open ended question on recent stressful events were talking to psychiatric patients, effects on personal life, presenting cases, and dealing with death and suffering. Relationships with consultants raised the strongest negative feelings, with 102 (34%) students finding these particularly stressful. Stress among medical students should be acknowledged and attempts made to alleviate it.


Women's preference for masculine traits is disrupted by images of male-on-female aggression.
Women's preferences for men's masculinized faces and voices were assessed after women (n = 331) were primed with images of male-on-male aggression, male-on-female aggression, pathogens, and neutral scenes. Male-on-male aggression and pathogen primes were associated with increased preference for masculine traits, but the same effect emerged in the neutral condition. We show the increased preference for masculine traits was due to repeated exposure to these traits, not the priming images themselves. Images of male-on-female aggression were an exception; these elicited feelings of disgust and anger appeared to disrupt the preference for masculinized traits. The results suggest women process men's facial and vocal traits as signals of aggressive potential and lose any preference for these traits with cues indicating men might direct this aggression toward them.


Hypercrosslinked porous polymer materials: design, synthesis, and applications.
Hypercrosslinked polymers (HCPs) are a series of permanent microporous polymer materials initially reported by Davankov, and have received an increasing level of research interest. In recent years, HCPs have experienced rapid growth due to their remarkable advantages such as diverse synthetic methods, easy functionalization, high surface area, low cost reagents and mild operating conditions. Judicious selection of monomers, appropriate length crosslinkers and optimized reaction conditions yielded a well-developed polymer framework with an adjusted porous topology. Post fabrication of the as developed network facilitates the incorporation of various chemical functionalities that may lead to interesting properties and enhance the selection toward a specific application. To date, numerous HCPs have been prepared by post-crosslinking polystyrene-based precursors, one-step self-polycondensation or external crosslinking strategies. The advent of these methodologies has prompted researchers to construct well-defined porous polymer networks with customized micromorphology and functionalities. In this review, we describe not only the basic synthetic principles and strategies of HCPs, but also the advancements in the structural and morphological study as well as the frontiers of potential applications in energy and environmental fields such as gas storage, carbon capture, removal of pollutants, molecular separation, catalysis, drug delivery, sensing etc.


Dietary inflammatory index and risk of first myocardial infarction; a prospective population-based study.
Chronic, low-grade inflammation is an established risk factor for cardiovascular disease. The inflammatory impact of diet can be reflected by concentrations of inflammatory markers in the bloodstream and the inflammatory potential of diet can be estimated by the dietary inflammatory index (DIITM), which has been associated with cardiovascular disease risk in some previous studies. We aimed to examine the association between the DII and the risk of first myocardial infarction (MI) in a population-based study with long follow-up. We conducted a prospective case-control study of 1389 verified cases of first MI and 5555 matched controls nested within the population-based cohorts of the Northern Sweden Health and Disease Study (NSHDS), of which the largest is the ongoing Västerbotten Intervention Programme (VIP) with nearly 100 000 participants during the study period. Median follow-up from recruitment to MI diagnosis was 6.4 years (6.2 for men and 7.2 for women). DII scores were derived from a validated food frequency questionnaire (FFQ) administered in 1986-2006. Multivariable conditional logistic regression models were used to estimate odds ratios (OR) and 95% confidence intervals (CI), using quartile 1 (most anti-inflammatory diet) as the reference category. For validation, general linear models were used to estimate the association between the DII scores and two inflammatory markers, high-sensitivity C-reactive protein (hsCRP) and interleukin 6 (IL-6) in a subset (n = 605) of the study population. Male participants with the most pro-inflammatory DII scores had an increased risk of MI [ORQ4vsQ1 = 1.57 (95% CI 1.21-2.02) P trend = 0.02], which was essentially unchanged after adjustment for potential confounders, including cardiovascular risk factors [ORQ4vsQ1 = 1.50 (95% CI 1.14-1.99), P trend = 0.10]. No association was found between DII and MI in women. An increase of one DII score unit was associated with 9% higher hsCRP (95% CI 0.03-0.14) and 6% higher IL-6 (95% CI 0.02-0.11) in 605 controls with biomarker data available. A pro-inflammatory diet was associated with an elevated risk of first myocardial infarction in men; whereas for women the relationship was null. Consideration of the inflammatory impact of diet could improve prevention of cardiovascular disease.


Reduced anterior hippocampal formation volume in hyponatremic schizophrenic patients.
Diminished hippocampal volume occurs in the anterior segment of some schizophrenic patients, and in the posterior segment in others. The significance of hippocampal pathology in general and these segmental differences in specific is not known. Several lines of evidence suggest anterior hippocampal pathology underlies the life-threatening hyponatremia seen in a subgroup of patients with schizophrenia; therefore our goal was to determine if this region was preferentially diminished in hyponatremic patients. We studied seven polydipsic hyponatremic, ten polydipsic normonatremic, and nine nonpolydipsic normonatremic schizophrenic inpatients, as well as 12 healthy controls. All underwent structural scanning on a high resolution (3.0 T) magnetic resonance imaging (MRI) scanner. Hippocampal formation, amygdala, and third ventricle volumes were manually traced in each subject. The hippocampus was divided at the posterior extent of the uncus, and all structural volumes were corrected for whole brain volume and other significant recognized factors (i.e., age, gender, height, parental education). Despite being overhydrated, anterior hippocampal formation volume was diminished in those with polydipsia and hyponatremia relative to each of the other three groups. Third ventricle volume was larger in this group than in healthy controls but similar to the two patient groups. Posterior hippocampal and amygdala volumes did not differ between groups. Other potential confounds (e.g., water imbalance) either had no effect or accentuated these differences. We conclude the anterior hippocampal formation is smaller in hyponatremic schizophrenic patients, thereby linking an important and objective clinical feature of schizophrenia to a neural pathway that can be investigated in animal models. The findings strengthen the hypothesis that anterior hippocampal formation pathology disrupts functional connectivity with other limbic structures in schizophrenia.


Relationship of oscillatory potential amplitude to A-wave slope over a range of flash luminances in normal subjects.
The effect of flash luminance on the relationship of oscillatory potential (OP) amplitude, as a measurement of inner retinal function, to a-wave slope, as a measurement of photoreceptor function, was evaluated in full-field electroretinograms recorded from normal subjects. The ratio of OP amplitude to a-wave slope was found to be independent of flash luminance over a 3000-fold luminance range. This finding raises the possibility that a reduction in the ratio of OP amplitude to a-wave slope in eyes with media opacities may be used as a sign of inner retinal malfunction. Selected cases are presented to illustrate application of this approach.


Cannabinoid Inheritance Relies on Complex Genetic Architecture.
Introduction: Understanding the inheritance of cannabinoid compounds in Cannabis sativa will facilitate effective crop breeding and careful regulation of controlled substances. The production of two key cannabinoids, Δ-9-tetrahydrocannabinol (THC) and cannabidiol (CBD), is partially controlled by two additive loci. Here, we present the first study to search for evidence of alternate genetic models describing the inheritance and expression of cannabinoids. Materials and Methods: Using an information-theoretic approach, we estimated composite genetic effects (CGEs) of four cultivars with pure CBD or pure THC chemotypes, their F1 and F2 hybrid progeny, to identify genetic models that explain cannabinoid inheritance patterns. We also estimated the effective number of genetic factors that control differences in cannabinoid concentration (THC, CBD, and cannabichromene [CBC]). Results: Unlike previous research, we note nonadditive components of cannabinoid inheritance. Concentration of THC is a polygenic trait (three to four genetic factors). Both additive and dominance CGEs best explained THC expression patterns. In contrast, cytoplasmic genomes and additive genes may influence CBD concentration. Maternal additive effects and additive genetic effects apparently influence CBC expression. Conclusions: Cannabinoid inheritance is more complex than previously appreciated; among other genetic effects, cytogenetic and maternal contributions may be undervalued influences on cannabinoid ratios and concentrations. Further research on the environmental sensitivity of cannabinoid production is advised.


"You can't cure it so you have to endure it": the experience of adaptation to diabetic renal disease.
In this study, the experience of adaptation to diabetic renal disease was examined from a phenomenological perspective. Twenty patients attending a diabetic renal clinic were interviewed in depth. Through the use of a template analysis approach, a set of strong themes relating to changes in lifestyle was identified: changes in the nature of involvement with the medical system, coping strategies, and hopes, fears, and expectations. Almost all participants attempted to construct a "good adaptation" in the face of the uncertainties surrounding their condition by adopting a stoic and fatalistic stance. This is discussed in the context of the claim that contemporary society holds emotional self-expression rather than stoical endurance to be the appropriate response to suffering.


A purely confirmatory replication study of structural brain-behavior correlations.
A recent 'crisis of confidence' has emerged in the empirical sciences. Several studies have suggested that questionable research practices (QRPs) such as optional stopping and selective publication may be relatively widespread. These QRPs can result in a high proportion of false-positive findings, decreasing the reliability and replicability of research output. A potential solution is to register experiments prior to data acquisition and analysis. In this study we attempted to replicate studies that relate brain structure to behavior and cognition. These structural brain-behavior (SBB) correlations occasionally receive much attention in science and in the media. Given the impact of these studies, it is important to investigate their replicability. Here, we attempt to replicate five SBB correlation studies comprising a total of 17 effects. To prevent the impact of QRPs we employed a preregistered, purely confirmatory replication approach. For all but one of the 17 findings under scrutiny, confirmatory Bayesian hypothesis tests indicated evidence in favor of the null hypothesis ranging from anecdotal (Bayes factor < 3) to strong (Bayes factor > 10). In several studies, effect size estimates were substantially lower than in the original studies. To our knowledge, this is the first multi-study confirmatory replication of SBB correlations. With this study, we hope to encourage other researchers to undertake similar replication attempts.


Ciguatera fish poisoning - New York City, 2010-2011.
During August 2010-July 2011, the New York City Department of Health and Mental Hygiene (DOHMH) received reports of six outbreaks and one single case of ciguatera fish poisoning (CFP), involving a total of 28 persons. CFP results from consumption of certain large, predatory, tropical reef fish that have bioaccumulated ciguatoxins (CTX). CFP is characterized by various gastrointestinal, cardiovascular, and neurologic symptoms. A prolonged period of acute illness can result, and the neurologic symptoms can last months, with variable asymptomatic and symptomatic periods. The first two outbreaks and the single case, involving 13 persons, were reported during August 6-September 13, 2010. DOHMH distributed a health alert in November 2010 requesting health-care providers be alert for CFP signs and symptoms. The health alert resulted in identification of 11 more cases that month and an additional two outbreaks involving four persons in July 2011. In comparison, only four CFP outbreaks, involving 21 persons total, had been reported in New York City (NYC) during the preceding 10 years (2000-2009). DOHMH's investigation revealed that 13 persons became ill after eating barracuda, and 15 became ill after eating grouper. Although specific and highly sensitive laboratory analyses can detect and confirm CTX in fish, no practical field tests are available for fish monitoring programs. CFP prevention depends on educating the public, seafood suppliers, and distributors about known CFP endemic areas and high-risk fish species. Traceback investigations of fish associated with outbreaks provide valuable information regarding fishing areas associated with CFP. Not all fish from CFP endemic areas are ciguatoxic, but persons who eat fish from endemic regions are at higher risk for CFP. If an illness is suspected to be CFP, public health authorities should be notified and informed of the case history for possible investigation and intervention measures.


Apoptosis is induced in cells with cytolytic potential by L-leucyl-L-leucine methyl ester.
Previous studies have demonstrated that the selective toxicity of leucyl-leucine methyl ester (Leu-Leu-OMe) for cytotoxic lymphocytes and myeloid cells is dependent on intracellular conversion to membranolytic metabolites by the acyl transferase activity of the granule enzyme dipeptidyl peptidase I (DPPI) that is enriched in these cells. The mechanism of cell death remained unclear, however, and was the subject of the experiments reported here. When human U937, HL60, or THP-1 myeloid tumor cell lines or murine CTLL-2 cells were treated with Leu-Leu-OMe, early release of both cytosolic 51Cr and soluble [3H]TdR labeled DNA fragments was observed, whereas antibody + C treatment of these cells caused only 51Cr release. Killing of U937 or THP-1 cells by incubation with the lysosomotropic amino acid methyl ester, Phe-OMe also induced only 51Cr release without evidence of DNA fragmentation. Preincubation with Zn2+, a known inhibitor of endonuclease activity prevented Leu-Leu-OMe-induced 51Cr or [3H]TdR release from these cell lines, but had no effect on antibody + C or Phe-OMe-induced 51Cr release. Zn2+ also prevented Leu-Leu-OMe-mediated killing of normal human CD16+ NK cells. Zn2+ had no inhibitory effect on Leu-Leu-OMe uptake or intracellular conversion to (Leu-Leu)n-OMe metabolites by these cell lines. Moreover, Zn2+ did not inhibit 51Cr release from nonnucleated E or nucleated U937 targets induced by extracellular production of DPPI-generated metabolites of Leu-Leu-OMe. Thus, killing of cytotoxic lymphocytes and myeloid cells by Leu-Leu-OMe appears to be dependent on generation of metabolites with membranolytic properties, but cell death induced by this process does not involve simple lysis of the plasma membrane. Rather, intracellular production of DPPI generated (Leu-Leu)n-OMe metabolites appears to trigger, an additional Zn(2+)-sensitive process that is associated with induction of apoptosis in cells with cytolytic potential.


Relationships between quantitative spinal cord MRI and retinal layers in multiple sclerosis.
To assess relationships between spinal cord MRI (SC-MRI) and retinal measures, and to evaluate whether these measures independently relate to clinical disability in multiple sclerosis (MS). One hundred two patients with MS and 11 healthy controls underwent 3-tesla brain and cervical SC-MRI, which included standard T1- and T2-based sequences and diffusion-tensor and magnetization-transfer imaging, and optical coherence tomography with automated segmentation. Clinical assessments included visual acuity (VA), Expanded Disability Status Scale, MS functional composite, vibration sensation threshold, and hip-flexion strength. Regions of interest circumscribing SC cross-sections at C3-4 were used to obtain cross-sectional area (CSA), fractional anisotropy (FA), perpendicular diffusivity (λ⊥), and magnetization transfer ratio. Multivariable regression assessed group differences and SC, retinal, and clinical relationships. In MS, there were correlations between SC-CSA, SC-FA, SC-λ⊥, and peripapillary retinal nerve fiber layer (pRNFL) (p = 0.01, p = 0.002, p = 0.001, respectively) after adjusting for age, sex, prior optic neuritis, and brain atrophy. In multivariable clinical models, when SC-CSA, pRNFL, and brain atrophy were included simultaneously, SC-CSA and pRNFL retained independent relationships with low-contrast VA (p = 0.04, p = 0.002, respectively), high-contrast VA (p = 0.06, p = 0.008), and vibration sensation threshold (p = 0.01, p = 0.05). SC-CSA alone retained independent relationships with Expanded Disability Status Scale (p = 0.001), hip-flexion strength (p = 0.001), and MS functional composite (p = 0.004). In this cross-sectional study of patients with MS, correlations exist between SC-MRI and retinal layers, and both exhibit independent relationships with clinical dysfunction. These findings suggest that the SC and optic nerve reflect ongoing global pathologic processes that supplement measures of whole-brain atrophy, highlighting the importance of combining measures from unique compartments to facilitate a thorough examination of regional and global disease processes that contribute to clinical disability in MS.