In bacteria with circular chromosomes, homologous recombination events can lead to the formation of chromosome dimers. In Escherichia coli, chromosome dimers are resolved by the addition of a crossover by two tyrosine recombinases, XerC and XerD, at a specific site on the chromosome, dif. Recombination depends on a direct contact between XerD and a cell division protein, FtsK, which functions as a hexameric double stranded DNA translocase. Here, we have investigated how the structure and composition of DNA interferes with Xer recombination activation by FtsK. XerC and XerD each cleave a specific strand on dif, the top and bottom strand, respectively. We found that the integrity and nature of eight bottom-strand nucleotides and three top-strand nucleotides immediately adjacent to the XerD-binding site of dif are crucial for recombination. These nucleotides are probably not implicated in FtsK translocation since FtsK could translocate on single stranded DNA in both the 5'-3' and 3'-5' orientation along a few nucleotides. We propose that they are required to stabilize FtsK in the vicinity of dif for recombination to occur because the FtsK-XerD interaction is too transient or too weak in itself to allow for XerD catalysis.

OBJECTIVE

The present study validated the abbreviated version of the World Health Organization Quality of Life (WHOQOL-BREF) questionnaire for general use in New Zealand.

METHODS

A random postal sample from the national electoral roll was used, and 808 questionnaires were returned. Psychometric properties of the instrument were assessed, including tests of the four-domain factor structure using confirmatory factor analysis and Rasch analysis.

RESULTS

Goodness-of-fit from the confirmatory factor analysis were good, and the overall conclusion of the Rasch analysis supported the confirmatory factor analysis (CFA) findings after dealing with problems of threshold ordering, local dependency, and differential item functioning (DIF).

CONCLUSIONS

The WHOQOL-BREF is valid for general use in New Zealand. In the future work, the WHOQOL-BREF domain scores should either be analyzed using non-parametric statistics or data should be fitted to the Rasch model to derive interval person estimates.

Dictyostelium discoidium cells express a family of cell surface cAMP receptors, and these G-protein-coupled receptors are each expressed with unique spatial and temporal patterns. One of these receptors, cAR2, is present during the postaggregative stages of development and our previous work suggests that it is preferentially expressed in prestalk cells. We report here the isolation of the promoter for carB, the gene which encodes cAR2. Using this fragment to generate a carB::lacZ, gene fusion construct, we investigated carB expression in detail. Expression is first detected at the tight aggregate stage and subsequently in a pattern reminiscent of the prestalk-specific gene ecmA. There are subtle differences, however, with, ecmA being expressed significantly in the anterior-like cells of the migrating pseudoplasmodium and in the basal disc and lower cup supporting the sorus during terminal development. carB is not expressed in any of these places. The presence of these different prestalk cell subtypes was confirmed by double indirect immunofluorescence using anti-cAR2 and anti-beta-galactosidase antibodies. While virtually all cAR2-expressing cells also express ecmA::lacZ, a substantial fraction of ecmA::lacZ-positive cells do not express cAR2. We also found the regulation of carB gene expression to differ from that of ecmA. carB expression is induced in vitro by extracellular cAMP, but surprisingly, not by DIF-1, a soluble molecule thought to be essential for the initiation of prestalk differentiation. Thus, cAR2 appears to be a cAMP receptor present on a restricted subset of prestalk cells and whose expression does not respond typically to the prestalk inducer DIF-1. DIF-1 sensitivity may, therefore, not be characteristic of all early prestalk differentiation.

The NF-κB family member p65 is central to inflammation and immunity. The purpose of this study was to identify and characterize evolutionary conserved genes modulating p65 transcriptional activity. Using an RNAi screening approach, we identified chaperonin containing TCP1 subunit η (CCTη) as a regulator of Drosophila NF-κB proteins, Dorsal and Dorsal-related immunity factor (Dif). CCTη was also found to regulate NF-κB-driven transcription in mammalian cells, acting in a promoter-specific context, downstream of IκB kinase (IKK). CCTη knockdown repressed IκBα and CXCL2/MIP2 transcription during the early phase of NF-κB activation while impairing the termination of CCL5/RANTES and CXCL10/IP10 transcription. The latter effect was associated with increased DNA binding and reduced p65 acetylation, presumably by altering the activity of histone acetyltransferase CREB-binding protein (CBP). We identified p65 lysines (K) 122 and 123 as target residues mediating the CCTη-driven termination of NF-κB-dependent transcription. We propose that CCTη regulates NF-κB activity in a manner that resolves inflammation.

Serotype O157:H7 is most frequently encountered among verotoxin-producing Escherichia coli. Most laboratories use MacConkey-sorbitol agar as a screening medium. Presumptive identification of sorbitol-negative colonies is then accomplished by latex agglutination or biochemical tests with serologic confirmation, which requires 18-36 hours for completion. This study attempted to detect E coli O157:H7 directly from stool specimens by direct immunofluorescence (DIF) antibody staining to provide quicker turnaround (< 2 hours). A total of 336 abnormal fecal samples (bloody, watery, semi-liquid, or mucoid) were examined by this method. Results were compared with those of culture. Direct immunofluorescence antibody staining of the direct fecal smear detected all isolates of E coli O157 that were recovered by culture, including nonmotile strains, strains possessing the H7 flagellar antigen, and one strain with a flagellar antigen other than H7. Optimum results were achieved when specimens were pretreated with 5% bleach and centrifugation. No false-negative results were obtained with bleach-pretreated stool samples.

OBJECTIVE

To assess whether an oral health-related quality of life (OHRQoL)measure showed differential item functioning (DIF) by ethnicity.

METHODS

A simple random sample of 12- and 13-year-old schoolchildren enrolled in the Taranaki District Health Board's school dental service, New Zealand. Each child (n = 430) completed the Child Perception Questionnaire (CPQ(11-14)) in the dental clinic waiting room, prior to a dental examination. The dataset included age, gender, ethnicity, and deprivation status. The general principle of the analytic plan was that equal scores from each CPQ(11-14) item were expected from both non-Mäori and Mäori groups regardless of their ethnic group. Ordinal logistic regression was performed. The dependent variables were the CPQ(11-14) items. The ethnicity group and each CPQ(11-14) domain score were the independent variables. Non-uniform DIF was assessed through adding an interaction term for each CPQ(11-14) sub-scale.

RESULTS

Non-uniform DIF was found in two items, one in the Functional Limitations sub-scale and another in the Social Well-being sub-scale. Uniform DIF was found in one item of the Emotional Well-being sub-scale.

CONCLUSIONS

Both non-uniform and uniform DIF by ethnicity was found in three of 37 items of the CPQ(11-14) questionnaire, showing it is important to perform DIF analysis when applying OHRQoL measures.

In this study, an item response theory-based differential functioning of items and tests (DFIT) framework (N. S. Raju, W. J. van der Linden, & P. F. Fleer, 1995) was applied to a Likert-type scale. Several differential item functioning (DIF) analyses compared the item characteristics of a 10-item satisfaction scale for Black and White examinees and for female and male examinees. F. M. Lord's (1980) chi-square and the extended signed area (SA) measures were also used. The results showed that the DFIT indices consistently performed in the expected manner. The results from Lord's chi-square and the SA procedures were somewhat varied across comparisons. A discussion of these results along with an illustration of an item with significant DIF and suggestions for future DIF research are presented.

OBJECTIVE

Pain is identified as the third largest health problem in the world, impacting sleep, physical functioning, and psychologic functioning. Pain has been viewed as a multidimensional experience influenced by biology, psychology, cultural conditioning, expectancies, and social contingencies. The aims of this study were to examine the psychometric properties and the differential item functioning (DIF) of the short-form McGill Pain Questionnaire (SF-MPQ) to test the cultural differences in verbally based pain expression.

METHODS

Data were drawn from the medical records of 134 Hispanic and 134 non-Hispanic White patients with pain. Cases were matched on total SF-MPQ score, sex, and age. Estimates of reliability and validity were assessed. Using a traditional moderated regression approach, each of the 15 items on the SF-MPQ was analyzed for DIF using matched moderated regression methodology.

RESULTS

Internal consistency was high: Hispanics 0.90 and non-Hispanics 0.89. Factor analysis revealed 2 components for each ethnic group. DIF analyses revealed statistically significant group differences for the 5 items (ie, throbbing, gnawing, aching, tiring-exhausting, and sickening) potentially due to cultural-linguistic factors. The remaining 10 items revealed no significant differences. Group differences were not accounted for by sex, age, marital status, education, pain location, or pain duration.

CONCLUSIONS

Test bias was relatively small (0.74) given the entire scale of the SF-MPQ (0 to 45), indicating that this measure seems to be used equivalently across these 2 groups. This study shows 1 method in evaluating the cross-cultural validity of pain assessment instruments, and contributes to the understanding of cross-cultural variability in item reporting on the SF-MPQ.

BACKGROUND

Differential item functioning (DIF) analyses are commonly used to evaluate health-related quality of life (HRQoL) instruments. There is, however, a lack of consensus as to how to assess the practical impact of statistically significant DIF results.

METHODS

Using our previously published ordinal logistic regression DIF results for the Fatigue scale of a HRQoL instrument as an example, the practical impact on a particular Norwegian clinical trial was investigated. The results were used to determine the difference in mean Fatigue scores assuming that the same trial was conducted in the UK. The results were then compared with published information on what would be considered a clinically important change in scores.

RESULTS

The item with the largest DIF effect resulted in differences between the mean English and Norwegian Fatigue scores that, although small, could be considered clinically important. Sensitivity analyses showed that larger differences were found for shorter scales, and when the proportions in each response category were equal.

CONCLUSIONS

Our scenarios suggest that translation differences in an item can result in small, but clinically important, differences at the scale score level. This is more likely to be problematic for observational studies than for clinical trials, where randomised groups are stratified by centre.

The finding that people of Chinese heritage tend to emphasize somatic rather than psychological symptoms of depression has frequently been discussed in the culture and mental health literature since the 1970s. Recent studies have confirmed that Chinese samples report more somatic and fewer psychological depression symptoms compared to "Western" samples. The question remains, however, as to whether or not these effects are attributable to variation in all the constituent symptoms or to a subset. If the latter, there is the additional possibility that some symptoms might show a divergent pattern. Such findings would have implications for how cultural variations in symptom presentation are interpreted, and would also inform the cultural study of affective experiences more broadly. The current study addressed these issues in Chinese (n = 175) and Euro-Canadian (n = 107) psychiatric outpatients originally described by Ryder et al. (2008). Differential item functioning (DIF) was used to examine whether specific somatic and psychological symptoms diverged from the overall patterns of cultural variation. Chi-square analyses were used to examine atypical somatic symptoms (e.g., hypersomnia), previously neglected in this literature. No DIF was observed for the typical somatic symptoms, but Euro-Canadians reported greater levels of atypical somatic symptoms, and showed higher rates of atypical depression. DIF was observed for psychological symptoms-the Chinese reported high levels of "suppressed emotions" and "depressed mood," relative to their overall psychological symptom reporting. Chinese outpatients also spontaneously reported "depressed mood" at similar levels as the Euro-Canadians, contrary to prevailing ideas about Chinese unwillingness to discuss depression. Overall, the findings provide a more nuanced picture of how culture shapes symptom presentation and point toward future studies designed to unpack cultural variation in narrower subsets of depressive symptoms.

We evaluated the efficacy, safety, and biological mechanisms of digoxin immune Fab (DIF) treatment of severe preeclampsia. Fifty-one severe preeclamptic patients were randomized in double-blind fashion to DIF ( N = 24) or placebo ( N = 27) for 48 hours. Primary outcomes were change in creatinine clearance (CrCl) at 24 to 48 hours and antihypertensive drug use. Serum sodium pump inhibition, a sequela of endogenous digitalis-like factors (EDLF), was also assessed. CrCl in DIF subjects was essentially unchanged from baseline versus a decrease with placebo (-3 +/- 10 and -34 +/- 10 mL/min, respectively, P = 0.02). Antihypertensive use was similar between treatments (46 and 52%, respectively, P = 0.7). Serum sodium pump inhibition was decreased with DIF compared with placebo at 24 hours after treatment initiation (least squares mean difference, 19 percentage points, P = 0.03). DIF appeared to be well tolerated. These results suggest DIF prevents a decline in renal function in severe preeclampsia by neutralizing EDLF. Sodium pump inhibition was significantly improved. Further research is warranted.

OBJECTIVE

Fatigue is one of the most common symptoms associated with cancer and its treatment. To obtain a more precise and flexible measure of fatigue, the EORTC Quality of Life Group has developed a computerized adaptive test (CAT) measure of fatigue. This is part of an ongoing project developing a CAT version of the widely used EORTC QLQ-C30 questionnaire.

METHODS

Based on the literature search and evaluations by experts and patients, 41 new fatigue items were developed (in addition to the three QLQ-C30 fatigue items). Psychometric properties of the items, including evaluations of dimensionality, fit to item response theory (IRT) model, and differential item functioning (DIF), were assessed in an international sample of cancer patients.

RESULTS

Responses were obtained from 1,321 cancer patients coming from eight countries. Factor analysis showed that 37 of the items could be included in a unidimensional model (RMSEA = 0.098, TLI = 0.995, CFI = 0.920). Of the 37 items, two were deleted because of poor fit to the IRT model forming the basis for the CAT, and one because of DIF between cancer sites.

CONCLUSIONS

We have established a 34-item fatigue bank allowing for more precise and flexible measurement of fatigue, while still being backward compatible with the QLQ-C30 fatigue scale.

BACKGROUND

Limited data exist regarding management of patients with a single brain lesion with extracranial disease due to non-small cell lung cancer (NSCLC).

METHODS

Eighty-eight consecutive patients with a single brain lesion from NSCLC in the presence of extracranial disease were treated with stereotactic radiosurgery (SRS) alone. Local control (LC), distant intracranial failure (DIF), overall survival (OS), and toxicity were assessed. The logrank test was used to identify prognostic variables.

RESULTS

Median OS was 10.6 months. One-year DIF was 61%; LC 89%. Treatments were delivered in 1-5 fractions to median BED10 = 60 Gy. Five patients developed radionecrosis. Factors associated with shortened OS included poor performance status (PS) (p = 0.0002) and higher Recursive Partitioning Analysis class (p = 0.017). For patients with PS 0, median survival was 22 months. DIF was associated with systemic disease status (progressive vs. stable) (p = 0.0001), as was BED (p = 0.021) on univariate analysis, but only systemic disease (p = 0.0008) on multivariate analysis.

CONCLUSIONS

This study identifies a patient population that may have durable intracranial control after treatment with SRS alone. These data support the need for prospective studies to optimize patient selection for up-front SRS and to characterize the impact of DIF on patients' quality of life.

Xer site-specific recombination functions in maintaining circular replicons in the monomeric state in Escherichia coli. Two recombinases of the bacteriophage lambda integrase family, XerC and XerD, are required for recombination at the chromosomal site, dif, and at a range of plasmid-borne sites. Xer recombination core sites contain the 11-base pair binding sites for each recombinase separated by a 6 to 8-base pair central region. We report that both XerC and XerD act as site-specific type I topoisomerases by relaxing supercoiled plasmids containing a dif site. Relaxation by either XerC or XerD occurs in the absence of the partner recombinase and requires only a single recombination core site. XerC or XerD relaxation activities are completely inhibited by the addition of the partner recombinase, providing that the DNA recognition sequence for the inhibiting partner is present.

This study used data from 3 sites to examine the invariance and psychometric characteristics of the Brief Symptom Inventory-18 across Black, Hispanic, and White mothers of 5th graders (N = 4,711; M = 38.07 years of age, SD = 7.16). Internal consistencies were satisfactory for all subscale scores of the instrument regardless of ethnic group membership. Mean and covariance structures analysis indicated that the hypothesized 3-factor structure of the instrument was not robust across ethnic groups. It provided a reasonable approximation to the data for Black and White women but not for Hispanic women. Tests for differential item functioning (DIF) were therefore conducted for only Black and White women. Analyses revealed no more than trivial instances of nonuniform DIF but more substantial evidence of uniform DIF for 3 of the 18 items. After having established partial strong factorial invariance of the instrument, latent factor means were found to be significantly higher for Black than for White women on all 3 subscales (somatization, depression, anxiety). In conclusion, the instrument may be used for mean comparisons between Black and White women.

OBJECTIVE

The problems of mood disorders are critical in people with epilepsy. Therefore, there is a need to validate a useful tool for the population. The Hospital Anxiety and Depression Scale (HADS) has been used on the population, and showed that it is a satisfactory screening tool. However, more evidence on its construct validity is needed.

METHODS

A total of 1041 people with epilepsy were recruited in this study, and each completed the HADS. Confirmatory factor analysis (CFA) and Rasch analysis were used to understand the construct validity of the HADS. In addition, internal consistency was tested using Cronbachs' α, person separation reliability, and item separation reliability. Ordering of the response descriptors and the differential item functioning (DIF) were examined using the Rasch models.

RESULTS

The HADS showed that 55.3% of our participants had anxiety; 56.0% had depression based on its cutoffs. CFA and Rasch analyses both showed the satisfactory construct validity of the HADS; the internal consistency was also acceptable (α=0.82 in anxiety and 0.79 in depression; person separation reliability=0.82 in anxiety and 0.73 in depression; item separation reliability=0.98 in anxiety and 0.91 in depression). The difficulties of the four-point Likert scale used in the HADS were monotonically increased, which indicates no disordering response categories. No DIF items across male and female patients and across types of epilepsy were displayed in the HADS.

CONCLUSIONS

The HADS has promising psychometric properties on construct validity in people with epilepsy. Moreover, the additive item score is supported for calculating the cutoff.

Intracellular pH (pHi) has been measured in Dictyostelium discoideum cells by 31P nuclear magnetic resonance. Ax2 cells, newly harvested from growth medium, maintained a pHi of 7.33 +/- 0.04 (17) at an extracellular pH ranging from 3.5 to 6.5. Below pH 3.5 the cells tend to lyse, whereas at pH values above 6.5 their pHi rises though they remain viable. pHi regulation in acid medium is not dependent on external Na+ or any other inorganic ion and so most probably involves the electrogenic plasma membrane proton pump. No significant change in pHi was detected during development through to the slug stage. Mature stalk cells gave a very acidic phosphate signal (pH less than or equal to 5.5) which was probably vacuolar in origin. Indirect experiments had suggested that pHi might regulate the development of Dictyostelium cells, with low pHi favouring stalk cell and high pHi favouring spore cell differentiation. In particular, two inhibitors of the plasma membrane proton pump, diethylstilbestrol and zearalenone, had been shown to be stalk cell inducers. In the present studies measurements of pHi of cells exposed to these inducers failed to detect the expected drop in pHi. In addition, DIF-1 (a low Mr factor), the natural inducer of stalk cell formation, caused, if anything, a slight alkalinization of the cells. Thus the original theory linking pHi and cell differentiation is not supported by these results and therefore appears to require some modification. Finally, extract experiments revealed the existence of two unidentified abundant phospho-compounds with resonant frequencies close to inorganic phosphate. The existence of these compounds can complicate the interpretation of spectra gained from living Dictyostelium cells.

Differential item functioning (DIF) is an important issue of interest in psychometrics and educational measurement. Several methods have been proposed in recent decades for identifying items that function differently between two or more groups of examinees. Starting from a framework for classifying DIF detection methods and from a comparative overview of the most traditional methods, an R package for nine methods, called difR, is presented. The commands and options are briefly described, and the package is illustrated through the analysis of a data set on verbal aggression.

OBJECTIVE

To assess, within the context of Rasch analysis, (1) the internal validity and reliability of the Berg Balance Scale (BBS) in a sample of rehabilitation patients with varied balance abilities; and (2) the comparability of the BBS measures across different neurologic diseases.

METHODS

Observational prospective study.

METHODS

Rehabilitation ward of an Italian district hospital.

METHODS

Consecutively admitted inpatients and outpatients (N=217); for 85 participants, data were collected both on admission and discharge, giving a total sample of 302 observations.

METHODS

Not applicable.

METHODS

BBS.

RESULTS

Most of the BBS items had to be rescored, and 2 items (static sitting and standing balance) had to be deleted, to attain adequate internal construct validity (χ(2)(24)=35.68; P=.059). The reliability of the Rasch-modified BBS (BBS-12) (total score, 0-35) was high (.957), indicating precision of measurement at the individual level. The analysis of differential item functioning (DIF) showed invariance of the item calibrations across patients' sex, age, and etiology. After adjusting for the possible effect of repeated measurements on person estimates, the analysis of DIF by timing of assessment confirmed the stability of the item hierarchy across time. A practical ruler was provided to convert item raw scores into Rasch estimates of balance ability.

CONCLUSIONS

This study supports the internal validity and reliability of the BBS-12 as a measurement tool independent of the etiology of the neurologic disease causing the balance impairment. In view of some sample-related issues and that not all possible etiologies encountered in the neurorehabilitation settings were tested, a larger multicenter study is warranted to confirm these findings.

OBJECTIVE

To evaluate the sensitivity to change, in patients who undergo vision rehabilitation, of the Veteran Affairs (VA) Low Vision Visual Functioning Questionnaire (LV VFQ-48), which was designed to measure the difficulty visually impaired persons have in performing daily activities and to evaluate vision rehabilitation outcomes.

METHODS

Before and after rehabilitation, the VA LV VFQ-48 was administered by telephone interview to subjects from five sites in the VA and private sector. Visual acuity of these subjects ranged from near normal to total blindness.

RESULTS

The VA LV VFQ exhibited significant differential item functioning (DIF) for 7 of 48 items (two mobility tasks, four reading tasks, and one distance-vision task). However, the DIF was small relative to baseline changes in item difficulty for all items. Therefore, the data were reanalyzed with the constraint that item difficulties do not change with rehabilitation, which assigns all changes to the person measure. Subjects in the inpatient Blind Rehabilitation Center (BRC) program showed the largest changes in person measures after vision rehabilitation (effect size = 1.9; t-test P < 0.0001). The subjects in the outpatient programs exhibited smaller changes in person measures after rehabilitation (effect size = 0.29; t-test P < 0.01). There was no significant change in person measures for the control group (test-retest before rehabilitation).

CONCLUSIONS

In addition to being a valid and reliable measure of visual ability, the VA LV VFQ-48 is a sensitive measure of changes that occur in visual ability as a result of vision rehabilitation. Patients' self-reports of the difficulty they experience performing daily activities measured with this instrument can be used to compute a single number, the person measure that can serve as an outcome measure in clinical studies. The VA LV VFQ-48 can be used to compare programs that offer different levels of intervention and serve patients across the continuum of vision loss.

The differentiation-inducing factor-1 (DIF-1) isolated from Dictyostelium discoideum is a potent antiproliferative agent that induces growth arrest and differentiation in mammalian cells in vitro. However, the specific target molecule(s) of DIF-1 has not been identified. In this study, we have tried to identify the target molecule(s) of DIF-1 in mammalian cells, examining the effects of DIF-1 and its analogs on the activity of some candidate enzymes. DIF-1 at 10-40 micro M dose-dependently suppressed cell growth and increased the intracellular cyclic AMP concentration in K562 leukemia cells. It was then found that DIF-1 at 0.5-20 micro M inhibited the calmodulin (CaM)-dependent cyclic nucleotide phosphodiesterase (PDE1) in vitro in a dose-dependent manner. Kinetic analysis revealed that DIF-1 acted as a competitive inhibitor of PDE1 versus the substrate cyclic AMP. Because DIF-1 did not significantly affect the activity of other PDEs or CaM-dependent enzymes and, in addition, an isomer of DIF-1 was a less potent inhibitor, we have concluded that PDE1 is a pharmacological and specific target of DIF-1.

Despite widespread assumptions about women's empowerment and agency in the Arab Middle East, psychometric research of these constructs is limited. Using national data from 6214 married women ages 16-49 who took part in the 2006 Egypt Labor Market Panel Survey, we applied factor analysis to explore and then to test the factor structure of women's agency. We then used multiple indicator multiple cause structural equations models to test for differential item functioning (DIF) by women's age at first marriage, a potential resource for women's agency. Our results confirm that women's agency in Egypt is multi-dimensional and comprised of their (1) influence in family decisions, including those reserved for men, (2) freedom of movement in public spaces, and (3) attitudes about gender, specifically violence against wives. These dimensions confirm those explored previously in selected rural areas of Egypt and South Asia. Yet, three items showed significant uniform DIF by women's categorical age at first marriage, with and without a control for women's age in years. Models adjusting for DIF and women's age in years showed that women's older age at first marriage was positively associated with the factor means for family decision-making and gender-violence attitudes, but not freedom of movement. Our findings reveal the value of our analytical strategy for research on the dimensions and determinants of women's agency. Our approach offers a promising model to discern "hierarchies of evidence" for social policies and programs to enhance women's empowerment.

Innate immunity consists of humoral and cellular components that play a vital role in regulation of defense responses to various pathogens in vertebrates and invertebrates. Recent studies have shown that Rel/DIF (dorsal-related immunity factor), Relish, STAT (signal transducer and activator of transcription) and CREB (cAMP response element-binding protein) transcription factor associated pathways are evolutionarily conserved across the animal kingdom. Although the primary role and general structure of the pathways in immunity have been revealed in many invertebrates, particularly arthropods, almost nothing is known about these pathways in the freshwater snail Biomphalaria glabrata, the intermediate host of the human blood fluke Schistosoma mansoni, which is a causative agent of human schistosomiasis. Given the central role of transcription factors (TF) in controlling expression of effector genes, understanding the role of a given TF is essential to obtaining insight into the general function of the corresponding signaling pathway. To better understand the immunity of B. glabrata, we investigated five homologues of TFs that have been shown to be associated with multiple prominent immune signaling pathways based on the considerable data reported from a wide phylogenetic range of animals. In this study we identified and characterized cDNAs of five TFs from B. glabrata, designated BgRelish, BgRel, BgSTAT1, BgSTAT2 and BgCREB, for the first time. Among the five TFs, Relish is first reported in Lophotrochozoa, one of three superphyla in Metazoa. Our identification of class I (BgRelish) and II (BgRel) NF-κB in B. glabrata suggests the two pathways, Toll-like receptor (TLR) and immune deficiency (IMD)-like pathways, are present in the superphylum Lophotrochozoa. Preliminarily expression studies indicate these TF-associated pathways may be involved in the snail's anti-schistosome response. This study not only advances our understanding of the snail's defenses, but also provides new perspectives about the evolution of animal immunity.

In the natural environment, days are generally warmer than the night, resulting in a positive day/night temperature difference (+DIF). Plants have adapted to these conditions, and when exposed to antiphase light and temperature cycles (cold photoperiod/warm night [-DIF]), most species exhibit reduced elongation growth. To study the physiological mechanism of how light and temperature cycles affect plant growth, we used infrared imaging to dissect growth dynamics under +DIF and -DIF in the model plant Arabidopsis (Arabidopsis thaliana). We found that -DIF altered leaf growth patterns, decreasing the amplitude and delaying the phase of leaf movement. Ethylene application restored leaf growth in -DIF conditions, and constitutive ethylene signaling mutants maintain robust leaf movement amplitudes under -DIF, indicating that ethylene signaling becomes limiting under these conditions. In response to -DIF, the phase of ethylene emission advanced 2 h, but total ethylene emission was not reduced. However, expression analysis on members of the 1-aminocyclopropane-1-carboxylic acid (ACC) synthase ethylene biosynthesis gene family showed that ACS2 activity is specifically suppressed in the petiole region under -DIF conditions. Indeed, petioles of plants under -DIF had reduced ACC content, and application of ACC to the petiole restored leaf growth patterns. Moreover, acs2 mutants displayed reduced leaf movement under +DIF, similar to wild-type plants under -DIF. In addition, we demonstrate that the photoreceptor PHYTOCHROME B restricts ethylene biosynthesis and constrains the -DIF-induced phase shift in rhythmic growth. Our findings provide a mechanistic insight into how fluctuating temperature cycles regulate plant growth.