Comparing Breast Screening Protocols: Inserting Catch Trials Does Not Improve Sensitivity over Double Screening.
Breast screening is an important tool for the early detection of breast cancers. However, tumours are typically present in less than 1% of mammograms. This low prevalence could cause radiologists to detect fewer tumours than they otherwise would, an issue known as the prevalence effect. The aim of our study was to investigate a novel breast screening protocol, designed to decrease the number of tumours missed by radiologists, without increasing their workload. We ran two laboratory-based experiments to assess the degree to which the novel protocol, called the catch trial (CT) protocol, resulted in greater sensitivity (d') than the double screener protocol (DS), currently utilised in Australia. In our first experiment we found evidence that the CT protocol resulted in a criterion shift relative to the DS protocol but the evidence that sensitivity was greater in the CT protocol relative to the DS protocol was less clear. A second experiment, using more realistic stimuli that were more representative of actual tumours, also failed to find convincing evidence that sensitivity was greater in the CT protocol than in the DS protocol. This experiment instead found that both the hit rate and the false alarm rate increased in the CT protocol relative to the DS protocol. So while there was again evidence that the CT protocol induced a criterion shift, the sensitivity appeared to be approximately the same in both protocols. Our results suggest the CT protocol is unlikely to result in an improvement in sensitivity over the DS protocol, so we cannot recommend that it be trialled in a clinical setting.


Docosahexaenoic acid and oleic acid induce altered DNA methylation of individual CpG loci in Jurkat T cells.
Docosahexaenoic acid (DHA, 22:6n-3) and oleic acid (18:1n-9) can alter the DNA methylation of individual CpG loci in vivo and in vitro, although the targeting mechanism is unknown. We tested the hypothesis that the targeting of altered methylation is associated with putative transcription factor response elements (pTREs) proximal to modified loci. Jurkat cells were treated with 22:6n-3 or 18:1n-9 (both 15 μM) for eight days and DNA methylation measured using the MethylationEPIC 850K array. 1596 CpG loci were altered significantly (508 hypermethylated) by 22:6n-3 and 563 CpG loci (294 hypermethylated) by 18:1n-9. 78 loci were modified by both fatty acids. Induced differential methylation was not modified by the PPARα antagonist GW6471. DNA sequences proximal to differentially methylated CpG loci were enriched in zinc-finger pTREs. These findings suggest that zinc-finger-containing transcription factors may be involved in targeting altered DNA methylation modifying processes induced by fatty acids to individual CpG loci.


How would serum 25(OH)D level change in patients with inflammatory bowel disease depending on intestinal mucosa vitamin D receptor (VDR) and vitamin D1-α hydroxylase (CYP27B1)?
To investigate how the serum 25-hydroxyvitamin D (25(OH)D) level change in patients with inflammatory bowel disease (IBD) and investigate the intestinal mucosa vitamin D receptor (VDR) and vitamin D1-α hydroxylase (CYP27B1) expressions. A total of 105 patients with IBD were enrolled in the present study, including 49 cases with ulcerative colitis (UC) and 56 cases with Crohn's disease (CD), compared with 45 healthy controls (CON) during the same period by testing the permeability of the intestinal mucosa. The expressions of VDR and CYP27B1 in the intestinal mucosa were detected, so as the serum endotoxin, tumor necrosis factor (TNF)-α, and 25(OH)D levels. The lactulose and mannitol absorption ratio (LMR) and serum endotoxin and TNF-α levels were significantly higher in the IBD group than in the CON group (p<0.05). The levels of LMR, endotoxin, and TNF-α were higher in the UC group than in the CD group, but 25(OH)D was lower (p<0.05). VDR in the IBD and UC groups was down-regulated when compared with the CON group (p<0.05), but there was no significance between them (p>0.05). CYP27B1 in the IBD and CD groups was significantly up-regulated compared with the CON group (p<0.05), with no significant difference between them (p>0.05). Patients with IBD exhibit vitamin D metabolism imbalance, lower serum 25(OH)D, and lower VDR expression, but higher CYP27B1 expression in the colonic mucosa. However, VDR and CYP27B1 cannot be used to distinguish UC and CD.


Greedy de novo motif discovery to construct motif repositories for bacterial proteomes.
Bacterial surfaces are complex systems, constructed from membranes, peptidoglycan and, importantly, proteins. The proteins play crucial roles as critical regulators of how the bacterium interacts with and survive in its environment. A full catalog of the motifs in protein families and their relative conservation grade is a prerequisite to target the protein-protein interaction that bacterial surface protein makes to host proteins. In this paper, we propose a greedy approach to identify conserved motifs in large sequence families iteratively. Each iteration discovers a motif de novo and masks all occurrences of that motif. Remaining unmasked sequences are subjected to the next round of motif detection until no more significant motifs can be found. We demonstrate the utility of the method through the construction of a proteome-wide motif repository for Group A Streptococcus (GAS), a significant human pathogen. GAS produce numerous surface proteins that interact with over 100 human plasma proteins, helping the bacteria to evade the host immune response. We used the repository to find that proteins part of the bacterial surface has motif architectures that differ from intracellular proteins. We elucidate that the M protein, a coiled-coil homodimer that extends over 500 A from the cell wall, has a motif architecture that differs between various GAS strains. As the M protein is known to bind a variety of different plasma proteins, the results indicate that the different motif architectures are responsible for the quantitative differences of plasma proteins that various strains bind. The speed and applicability of the method enable its application to all major human pathogens.


Matrix coating assisted by an electric field (MCAEF) for enhanced tissue imaging by MALDI-MS.
A novel technique, termed matrix coating assisted by an electric field (MCAEF), for enhancing tissue imaging by matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) was developed in this study. In this technique a static and uniform electric field is applied to sliced tissue sections during matrix spray-coating, resulting in the enrichment of positively or negatively chargeable analytes in the MALDI matrix layer. Experimental results show that MCAEF not only increased the sensitivity of lipid and protein detection across the board in the subsequent MALDI-MS analyses, but also resulted in successful imaging of a larger number of analytes. MALDI imaging enhancement with MCAEF was observed for various tissues (rat liver, rat brain, and porcine adrenal gland) and with different MALDI matrices (e.g., quercetin, 2-mercaptobenzothiazole, dithranol, 9-aminoacridine, and sinapinic acid) and the sensitivity increases were independent of the solvent compositions and pH values of the matrix solutions. Taking rat brain as an example, MCAEF led to the on-tissue detection and imaging of 648 identified lipids by combining positive and negative ion detection by MALDI-Fourier transform ion cyclotron resonance MS and with quercetin as the matrix, as compared to only 344 lipids without MCAEF. For protein imaging, up to 232 protein signals were successfully detected in rat brain tissue sections by MALDI-time-of-flight MS within a mass range of 3500 to 37 000 Da, as compared to 119 without MCAEF. MCAEF also enabled the detection of higher molecular-weight proteins. These results demonstrate the advantages of MCAEF for overall performance improvements in MALDI imaging and we believe that this technique has the potential to become a standard practice for MALDI tissue imaging.


Anatomically safe and minimally invasive transcrestal technique for procurement of autogenous cancellous bone graft from the mid-iliac crest.
Open iliac bone harvesting techniques can result in significant complications and residual morbidity. In reconstructive procedures where a small volume of autogenous cancellous bone graft is required, a minimally invasive technique for bone harvesting applied at the mid-iliac crest has been deemed satisfactory. We sought to assess the application of a well-established surgical technique to procure adequate volume of autogenous cancellous iliac bone graft with minimal trauma to adjacent structures. We retrospectively reviewed the cases of patients who underwent a minimally invasive transcrestal mid-iliac bone graft procurement technique between May 2003 and December 2007. The technique was performed using a 3.5-mm Steinmann pin as a trocar and a 4.5-mm AO drill sleeve as a trephine. We administered a questionnaire, either in the clinic or by mail, to assess a number of parameters, including postoperative pain, dysthesia, parasthesia, status of the donor site wound and patient satisfaction. Of the 37 consecutive patients who underwent the procedure, data from 26 patients were available for assessment. Donor site pain resolved within a few days of the surgery, and none of the patients experienced symptoms of chronic pain. At the final review, none of the patients reported any unpleasant signs and symptoms related to the residual scar. We recommend that the described minimally invasive trephine method be used when a small cancellous bone graft is needed. We found that patient morbidity was significantly lower with the trephine harvest technique than with open bone harvesting methods at the anterior iliac crest.


BDNF variant Val66Met interacts with estrous cycle in the control of hippocampal function.
Natural fluctuations in circulating estradiol are associated with behavioral changes, including severe disturbances in mood and cognition in some women. Common genetic variation in some of the molecular mediators of estradiol effects on these behaviors, in brain regions such as the hippocampus, may explain individual variation in estradiol effects on behavior. We tested whether the common human variant BDNF Val66Met interacts with estradiol in the control of hippocampal function in cycling female mice homozygous for the wild-type Val or BDNF Met variant. BDNF Met increased anxiety behavior, impaired memory, and increased expression of BDNF and its receptor TrkB in the hippocampal formation. BDNF Met also dramatically altered the fluctuation of spatial memory, hippocampal Akt phosphorylation, and PSD-95 protein expression across the estrous cycle. The variant BDNF Val66Met should therefore be considered as a strong candidate for mediating genetic differences in ovarian steroid-related behavioral changes and disorders.


Intestinal microbiology in early life: specific prebiotics can have similar functionalities as human-milk oligosaccharides.
Human milk is generally accepted as the best nutrition for newborns and has been shown to support the optimal growth and development of infants. On the basis of scientific insights from human-milk research, a specific mixture of nondigestible oligosaccharides has been developed, with the aim to improve the intestinal microbiota in early life. The mixture has been extensively studied and has been shown to be safe and to have potential health benefits that are similar to those of human milk. The specific mixture of short-chain galacto-oligosaccharides and long-chain fructo-oligosaccharides has been found to affect the development of early microbiota and to increase the Bifidobacterium amounts as observed in human-milk-fed infants. The resulting gut ecophysiology is characterized by high concentrations of lactate, a slightly acidic pH, and specific short-chain fatty acid profiles, which are high in acetate and low in butyrate and propionate. Here, we have summarized the main findings of dietary interventions with these specific oligosaccharides on the gut microbiota in early life. The gut ecophysiology in early life may have consequences for the metabolic, immunologic, and even neurologic development of the child because reports increasingly substantiate the important function of gut microbes in human health. This review highlights major findings in the field of early gut colonization and the potential impact of early nutrition in healthy growth and development.


The impact of temporary deferrals on future blood donation behavior across the donor life cycle.
Donor retention is essential for blood banks because acquiring new donors is more expensive than retaining existing ones. Previous studies show that the temporary deferral of donors negatively impacts future donation likelihood. In this study, we analyze the impact of temporary deferrals on future donation behavior while correcting for potential endogeneity, depending on the level of donor experience and number of previous deferrals. We use data from more than 123,000 whole blood donors of the Austrian Red Cross over a period of 5.5 years. We estimate logit models to analyze how a deferral affects future donation behavior while controlling for potential selection biases because donors are not deferred randomly. We control for gender, blood type, years since first donation, and number of previous donations and deferrals. We analyze the direct deferral effect, its interaction with donor experience, and the number of previous deferrals. Our results confirm that temporary deferrals hurt future donation behavior. This effect varies with donor experience and the number of previous deferrals. The effect is weaker with a higher number of previous donations and is stronger with a higher number of previous deferrals. The results suggest that donors learn to cope with deferrals the more they donate. However, the negative effect of deferrals amplifies over time, and each additional deferral decreases donation likelihood. Blood banks that seek to overcome the negative effect of deferrals should be aware that this effect varies with donor experience and with the number of previous deferrals. Our results suggest that blood banks should focus on early-stage donors who are deferred because the negative deferral effect is stronger for more experienced donors. At the same time, blood banks should be careful with donor groups who have experienced deferrals in the past because every additional deferral demotivates future donation behavior. Overall, researchers should be careful to correct for endogeneity because our results suggest that ignoring these effects could lead to substantial underestimation of the negative deferral effect.


Bilateral submandibular salivary gland swelling--a report of chronic sialodochitis with eosinophilia.
A case of bilateral swelling of the submandibular salivary glands is presented. The histopathological features were diffuse periductal sclerosis with a lymphoplasmacytic infiltrate which was rich in eosinophils. Some acinar atrophy was seen. Salivary ducts showed mucous and squamous prosoplasia, with focal inspissated mucin. Numerous dilated and congested blood vessels were prominent throughout the stroma. These features overlapped with those of Kimura's disease and angiolymphoid hyperplasia with eosinophils. The features of these conditions and a differential diagnosis are discussed.


Pulmonary rehabilitation: summary of an evidence-based guideline.
Pulmonary rehabilitation has emerged as a standard of care for patients with chronic lung disease, based on a growing body of scientific evidence. Over recent decades, several organizations have championed pulmonary rehabilitation and developed comprehensive statements, practice guidelines, and evidence-based guidelines. Documenting the scientific evidence underlying clinical practice has been important in overcoming skepticism and convincing health professionals, health-care institutions, third-party payers, and regulatory agencies to support pulmonary rehabilitation programs. The literature on pulmonary rehabilitation has increased substantially and provided justification for including pulmonary rehabilitation in practice guidelines for chronic obstructive pulmonary disease and other chronic lung diseases. Therefore, the American College of Chest Physicians and the American Association of Cardiovascular and Pulmonary Rehabilitation decided to update their 1997 guidelines with a systematic, evidence-based review of the literature since the previous review. The panel updated prior topics and recommendations and reviewed new topics. Recommendations were given for outcomes of comprehensive pulmonary rehabilitation programs, including lower-extremity exercise training, dyspnea, health-related quality of life, health-care utilization, survival, psychosocial outcomes, and long-term benefits. Additional topics include the duration of pulmonary rehabilitation, post-rehabilitation maintenance strategies, intensity of aerobic exercise training, strength training, anabolic drugs, upper-extremity training, inspiratory-muscle training, education, psychological and behavioral components, oxygen supplementation, noninvasive ventilation, nutrition supplementation, rehabilitation for patients with disorders other than chronic obstructive pulmonary disease, and future pulmonary rehabilitation research. These guidelines provide an excellent summary of the recent literature and further strengthen the scientific basis of pulmonary rehabilitation.


What's hot, what's new at WTC--clinical science.
More than 3000 abstracts of innovative and exciting findings, covering the whole field of organ transplantation, were presented at the World Transplant Congress 2014. Key areas of presentations across all organs and tissues included HLA antibodies, antibody-mediated rejection, living donation, immunosuppression, organ perfusion and surgical procedures. In addition, cutting edge science and future perspectives were presented in state-of-the-art lectures. This review will present highlights of this meeting and demonstrate strength and success of clinical sciences in transplantation.


Recognition of sequence-directed DNA structure by the Klenow fragment of DNA polymerase I.
Time-resolved fluorescence spectroscopy was used to investigate the influence of sequence-directed DNA structure upon the interaction between the Klenow fragment of DNA polymerase I and a series of defined oligonucleotide primer/templates. 17/27-mer (primer/template) oligonucleotides containing a dansyl fluorophore conjugated to a modified deoxyuridine residue within the primer strand were used as substrates for binding to Klenow fragment. The time-resolved fluorescence anisotropy decay of the dansyl probe was analyzed in terms of two local environments, either solvent-exposed or buried, corresponding to primer/templates positioned with the primer 3' terminus in the polymerase site or the 3'-5' exonuclease site of the enzyme, respectively. Equilibrium constants for partitioning of DNA between the two sites were evaluated from the anisotropy decay data for primer/templates having different (A + T)-rich sequences flanking the primer 3' terminus. Primer/templates with AAAATG/TTTTAC and CGATAT/GCTATA terminal sequences (the nucleotides on the left refer to the last six bases at the 3' end of the primer, and the nucleotides on the right are the corresponding bases in the template) were bound mostly at the polymerase site. The introduction of single mismatches opposite the primer 3' terminus of these DNA substrates increased their partitioning into the 3'-5' exonuclease site, in accord with the results of an earlier study [Carver, T.E., Hochstrasser, R.A., and Millar, D.P. (1994) Proc. Natl. Acad. Sci. U.S.A. 91, 10670-10674]. In contrast, a primer/template with the terminal sequence CAATTT/GTTAAA, containing an A-tract element AATTT, exhibited a surprising preference for binding at the 3'-5' exonuclease site, despite the absence of mismatched bases in the DNA substrate. Interruption of the A-tract with a single AG step, to give the terminal sequence CAGTTT/GTCAAA, reversed the effect of the A-tract, causing the DNA to partition in favor of the polymerase site. Moreover, the presence of a single mismatch opposite the primer 3' terminus was also sufficient to reverse the effect of the A-tract, resulting in a distribution of DNA between polymerase and 3'-5' exonuclease sites that was similar to that observed for the other mismatched DNA substrates. Taken together, these results suggest that the A-tract adopts an unusual conformation that is disruptive to binding at the polymerase site. The effect of the A-tract on binding of DNA to the polymerase site is discussed in terms of the unusual helix structural parameters associated with these sequence elements and the difference between the local geometry of the A-tract and the conformation adopted by duplex DNA within the polymerase cleft. The results of this study show that in addition to base mismatches, Klenow fragment can also recognize irregularities in the helix geometry of perfectly base-paired DNA.


Molecular and biochemical characterization of a heat-labile cytotonic enterotoxin from Aeromonas hydrophila.
We report herein the DNA sequence analysis of the heat-labile cytotonic enterotoxin gene (alt) from Aeromonas hydrophila and the biological function of the purified hyperproduced toxin (Alt). One large open-reading frame (ORF), comprised of 1104 bp, was detected at positions 804 to 1907 bp on a 4.0-kb Sa/l DNA fragment from Aeromonas. This ORF encodes for a protein having 368 amino acids (aa) with a computed molecular weight of 38 kDa. The aa sequence of the first 15 NH2-terminal residues of the mature native Alt from A. hydrophila matched with the DNA-derived aa sequence of the alt gene expressed in E. coli starting at position 19, which was leucine. The first 18aa residues of the Alt represented a putative signal sequence with alanine at its carboxy terminus. A BLAST search of the entire database showed 45-51% identity of the Alt, starting at position 158 with the carboxy half of the phospholipase C (PLC) and lipase from A. hydrophila; however, the purified Alt had no lipase/PLC activity. The alt gene was hyperexpressed using gene fusion expression vector systems, and the recombinant Alt exhibited a size of 35-40 kDa. The pure recombinant Alt elongated Chinese hamster ovary cells and elicited fluid secretion in rats ligated intestinal loops, indicating its enterotoxicity. Immunization of mice with recombinant Alt resulted in a reduced fluid secretory response when challenged with Aeromonas. The biological activity of the recombinant Alt in E. coli was about 10-fold less, compared to native Alt from Aeromonas, indicating differential processing of the toxin. The antibodies to native Alt neutralized the biological activity of the recombinant toxin, and these antibodies reacted with the same specificity to the native and recombinant Alt in immunoblots. The role of cyclic adenosine monophosphate and prostaglandins in causing a fluid secretory response by Alt also was demonstrated.


A single-cell transcriptional roadmap for cardiopharyngeal fate diversification.
In vertebrates, multipotent progenitors located in the pharyngeal mesoderm form cardiomyocytes and branchiomeric head muscles, but the dynamic gene expression programmes and mechanisms underlying cardiopharyngeal multipotency and heart versus head muscle fate choices remain elusive. Here, we used single-cell genomics in the simple chordate model Ciona to reconstruct developmental trajectories forming first and second heart lineages and pharyngeal muscle precursors and characterize the molecular underpinnings of cardiopharyngeal fate choices. We show that FGF-MAPK signalling maintains multipotency and promotes the pharyngeal muscle fate, whereas signal termination permits the deployment of a pan-cardiac programme, shared by the first and second heart lineages, to define heart identity. In the second heart lineage, a Tbx1/10-Dach pathway actively suppresses the first heart lineage programme, conditioning later cell diversity in the beating heart. Finally, cross-species comparisons between Ciona and the mouse evoke the deep evolutionary origins of cardiopharyngeal networks in chordates.


Extrafoveal photostress recovery test in glaucoma and idiopathic central serous chorioretinopathy.
A photostress recovery test was designed to differentiate macular diseases from optic nerve disorders, but recently an abnormal recovery time was reported in glaucoma. The purpose of this study was to search for the difference in abnormality of the photostress recovery test between glaucoma and idiopathic central serous chorioretinopathy (ICSC). This study involved 21 normal subjects, 14 patients, with ICSC and 10 patients with primary open angle glaucoma (POAG). A scanning laser ophthalmoscope (SLO) was used with microperimetry for bleaching the test point and measuring the recovery of sensitivity. Photostress recovery time (SLO-PSRT) could be measured at extrafoveal points outside and inside the affected area. The initial sensitivity change and the time constant of recovery after bleaching were calculated by fitting an exponential equation to the data. In normal subjects, neither the initial sensitivity change nor the time constant were correlated with the location of the test point. In 14 patients with ICSC, the initial sensitivity change in the detached area was significantly smaller than that in the unaffected area which was not significantly different from that in the age matched normal subjects. The time constant in the detached area was significantly longer than that in the unaffected area, which was not significantly different from that in the normal subjects. In 10 patients with POAG, the initial sensitivity change inside and outside the scotoma was not significantly different from that of age matched normal subjects. The time constant inside the scotoma was significantly longer than that outside the scotoma, which was not significantly different from that of the age matched normal subjects. Both ICSC and POAG showed a prolonged time constant of recovery, but the initial sensitivity change was reduced only in ICSC. The difference in our results between ICSC and POAG may be caused by the difference of the retinal pathology. Further, the SLO-PSRT is very useful when the lesion is located outside the fovea.


Lung transplantation for high-risk patients with idiopathic pulmonary fibrosis.
Survival for patients with idiopathic pulmonary fibrosis (IPF) and high lung allocation score (LAS) values may be significantly reduced in comparison to those with lower LAS values. To evaluate outcomes for high-risk IPF patients as defined by LAS values ≥46 (N=42) versus recipients with LAS values <46 (N=89). We retrospectively reviewed records of 131 consecutive patients with IPF who received lung transplants at our institution between 1999 and 2013. The mean LAS was significantly higher (59.5, interquartile range 43.9-75.9 vs. 39.3, interquartile range 37.7-44.3; p<0.01) for the high-risk cohort. The higher LAS cohort had significantly lower percent predicted forced vital capacity (FVC) versus recipients with LAS <46 (41.3±14.1% vs. 53.2±16.2%; p<0.01) and required more supplemental oxygen (7±5 vs. 4±2 L/min, p<0.01) prior to transplant versus recipients with LAS <46. Although the incidence of early post-LTX pulmonary complications was increased for the higher LAS group versus recipients with LAS <46, 30-day mortality and actuarial survival did not differ between the two cohorts. Although lung transplantation in patients with IPF and high LAS values is associated with increased risk of early post-transplant complications, long-term post-transplant survival for our high-LAS cohort was equivalent to that for the lower LAS recipients.


MVnet: automated time-resolved tracking of the mitral valve plane in CMR long-axis cine images with residual neural networks: a multi-center, multi-vendor study.
Mitral annular plane systolic excursion (MAPSE) and left ventricular (LV) early diastolic velocity (e') are key metrics of systolic and diastolic function, but not often measured by cardiovascular magnetic resonance (CMR). Its derivation is possible with manual, precise annotation of the mitral valve (MV) insertion points along the cardiac cycle in both two and four-chamber long-axis cines, but this process is highly time-consuming, laborious, and prone to errors. A fully automated, consistent, fast, and accurate method for MV plane tracking is lacking. In this study, we propose MVnet, a deep learning approach for MV point localization and tracking capable of deriving such clinical metrics comparable to human expert-level performance, and validated it in a multi-vendor, multi-center clinical population. The proposed pipeline first performs a coarse MV point annotation in a given cine accurately enough to apply an automated linear transformation task, which standardizes the size, cropping, resolution, and heart orientation, and second, tracks the MV points with high accuracy. The model was trained and evaluated on 38,854 cine images from 703 patients with diverse cardiovascular conditions, scanned on equipment from 3 main vendors, 16 centers, and 7 countries, and manually annotated by 10 observers. Agreement was assessed by the intra-class correlation coefficient (ICC) for both clinical metrics and by the distance error in the MV plane displacement. For inter-observer variability analysis, an additional pair of observers performed manual annotations in a randomly chosen set of 50 patients. MVnet achieved a fast segmentation (<1 s/cine) with excellent ICCs of 0.94 (MAPSE) and 0.93 (LV e') and a MV plane tracking error of -0.10 ± 0.97 mm. In a similar manner, the inter-observer variability analysis yielded ICCs of 0.95 and 0.89 and a tracking error of -0.15 ± 1.18 mm, respectively. A dual-stage deep learning approach for automated annotation of MV points for systolic and diastolic evaluation in CMR long-axis cine images was developed. The method is able to carefully track these points with high accuracy and in a timely manner. This will improve the feasibility of CMR methods which rely on valve tracking and increase their utility in a clinical setting.


Identifying women with dense breasts at high risk for interval cancer: a cohort study.
Twenty-one states have laws requiring that women be notified if they have dense breasts and that they be advised to discuss supplemental imaging with their provider. To better direct discussions of supplemental imaging by determining which combinations of breast cancer risk and Breast Imaging Reporting and Data System (BI-RADS) breast density categories are associated with high interval cancer rates. Prospective cohort. Breast Cancer Surveillance Consortium (BCSC) breast imaging facilities. 365,426 women aged 40 to 74 years who had 831,455 digital screening mammography examinations. BI-RADS breast density, BCSC 5-year breast cancer risk, and interval cancer rate (invasive cancer ≤12 months after a normal mammography result) per 1000 mammography examinations. High interval cancer rate was defined as more than 1 case per 1000 examinations. High interval cancer rates were observed for women with 5-year risk of 1.67% or greater and extremely dense breasts or 5-year risk of 2.50% or greater and heterogeneously dense breasts (24% of all women with dense breasts). The interval rate of advanced-stage disease was highest (>0.4 case per 1000 examinations) among women with 5-year risk of 2.50% or greater and heterogeneously or extremely dense breasts (21% of all women with dense breasts). Five-year risk was low to average (0% to 1.66%) for 51.0% of women with heterogeneously dense breasts and 52.5% with extremely dense breasts, with interval cancer rates of 0.58 to 0.63 and 0.72 to 0.89 case per 1000 examinations, respectively. The benefit of supplemental imaging was not assessed. Breast density should not be the sole criterion for deciding whether supplemental imaging is justified because not all women with dense breasts have high interval cancer rates. BCSC 5-year risk combined with BI-RADS breast density can identify women at high risk for interval cancer to inform patient-provider discussions about alternative screening strategies. National Cancer Institute.


Tools for analyzing cell shape changes during chemotaxis.
Chemotaxis refers to the ability of cells to sense the direction of external chemical gradients and respond by migrating towards the source. A thorough understanding of the chemotactic response of amoebae and neutrophils requires careful quantification of the cell shape changes observed during cell movement. The stochastic nature of this response calls for a statistical characterization of cellular morphology and this requires the processing of large data sets. For this reason, automatic image analysis algorithms are highly desirable and are becoming increasingly available. These usually include a combination of techniques from image segmentation, morphological transformations, as well as the incorporation of numerical algorithms and physical models. Here we review recent developments in the tracking and understanding of motile chemotaxing cells, with a particular emphasis on the description of pseudopodial activity in chemotactic Dictyostelium cells.


Electroacupuncture inhibits CB1 upregulation induced by ethanol withdrawal in mice.
CB1R play a role in alcohol withdrawal and in some effects of acupuncture. Interestingly, acupuncture has been used to alleviate alcohol withdrawal. Here, we investigated electroacupuncture (EA) effects during ethanol withdrawal on CB1R immunoreactivity. Male Swiss mice were daily injected with ethanol (2g/kg, i.p) (EtOH group), for 21 days. EA was performed daily during 4 days of ethanol withdrawal. The stimuli of 2 or 100 Hz were provided in two acupoints combination: Ea1 [(ST-36/Zusanli) and (PC-6/Neiguan)] or Ea2 [(DU-14/Dazhui) and (DU-20/Baihui)]. The specificity of the acupoints were assessed by the inclusion of three additional groups, Ea3 [(ST 25/Tianshu - acupoints used to other non-related disorders)], Sham1 and Sham2 (transdermic stimulation nearly to the respective acupoints). EtOH group were only handled during withdrawal and Saline group was chronically treated with Saline and handled similarly to EtOH group. One day after withdrawal the animals were perfused and their brains processed for immunohistochemistry. There was an increase of CB1R in the prefrontal cortex, striatum, hippocampus, amygdala and ventral tegmental area. The procedures used in the 2HzEa1 and 100HzEa2 groups were the most effective and specific to inhibit this CB1R upregulation. Therefore, EA inhibits CB1R upregulation seen in ethanol withdrawn mice. The specificity of acupoints stimulation depends of the encephalic nuclei, acupoints association and frequency of stimulation.


Interpersonal needs in middle adolescents: companionship, leadership and intimacy.
Recent research suggests that, along with identity, intimacy is an important developmental construct during adolescence. Are there gender differences in current society regarding intimacy development? Two hundred and seven middle adolescents (70 males and 137 females) were measured using Schutz's (1958) Fundamental Interpersonal Relationship Inventory (FIRO). The FIRO is a self-report survey which assesses the subject's perceived expressions and perceived desires in three categories of interpersonal relationships: Inclusion (companionship), control (leadership), and affection (intimacy). Results indicated that there were differences in expression of inclusion, control, and affection, and desire for inclusion and affection. A second analysis addressed the perceived ranking in importance of the three interpersonal categories measured. Males ranked control expressed highest and affection desired lowest; females ranked affection desired as highest and control expressed lowest. Both groups ranked inclusion desired and expressed as moderate. The current research suggests that gender differences in the development of intimacy may occur as early as middle adolescence.


Is Free Running Asthma Screening Test still useful?
The aim of this study is to evaluate the usefulness of Free Running Asthma Screening Test (FRAST) as a method that may help the diagnosis of exercise-induced bronchospasm (EIB) in children from a rural area and to compare the results with their history of asthma. A FRAST challenge took place at 9 schools. It lasted for 6 minutes and pupils performed it properly with the help of an athlete. The best out of three Peak Expiratory Flow Rate (PEFR) efforts was recorded before and after challenge, using a mini-Wright peak-flow-meter. A more than 15% drop in PEFR was considered an indication of EIB. A questionnaire defining asthmatic symptoms had been distributed and already completed by their parents. The exercise was successfully performed by 268 children. Eleven children presented a fall of PEFR of at least 15%, five or ten minutes after the exercise; among these, only three boys had a history of asthma, while the remaining 8 children were undiagnosed. No difference regarding age, gender or locations' altitude was found. FRAST can be of great use in the diagnosis of EIB in areas that lack proper tools and specialized personnel. A poor correlation between self-reported symptoms and the results of exercise challenges was confirmed.


A reevaluation of CD22 expression in human lung cancer.
CD22 is a transmembrane glycoprotein expressed by mature B cells. It inhibits signal transduction by the B-cell receptor and its coreceptor CD19. Recent reports indicate that most human lung cancer cells and cell lines express CD22, making it an important new therapeutic target for lung cancer. The objective of our studies was to independently validate these results with the goal of testing the efficacy of our CD22 immunotoxins on lung cancer cell lines. As determined by quantitative real-time PCR analysis, we found that levels of CD22 mRNA in a panel of human lung cancer cell lines were 200 to 60,000-fold lower than those observed in the human CD22(+) Burkitt lymphoma cells, Daudi. Using flow cytometry with a panel of CD22 monoclonal antibodies and Western blot analyses, we could not detect surface or intracellular expression of CD22 protein in a panel of lung cancer cell lines. In addition, the in vitro proliferation of the lung tumor cell lines was not affected by either CD22 antibodies or our highly potent anti-CD22 immunotoxin. In contrast, CD22(+) Daudi cells expressed high levels of CD22 mRNA and protein, and were sensitive to our CD22 immunotoxin. Importantly, primary non-small cell lung cancers from more than 250 patient specimens did not express detectable levels of CD22 protein as assessed by immunohistochemistry. We conclude that CD22 is not expressed at measurable levels on the surface of lung cancer cells, and that these cells cannot be killed by anti-CD22 immunotoxins.


[Mortality for diarrheic disease in Mexico: problem of accessibility or quality of care?].
To evaluate the access to health services and the process of care provided to children under five years old who died from acute diarrhea (AD). In four states of Mexico, a cross-sectional study was carried out. Deaths from AD that occurred in the lapse of one year were included. Information about clinical characteristics of the illness, process of care and access to the health facilities were obtained through verbal autopsy. Descriptive analysis and comparison of data between rural and urban communities were performed. Five-hundred and fifty three deaths were analysed. Most of the children were not enrolled in a public care health system. The main children's characteristics were the following: residence in rural communities; age less than one year; died at home or had short illness duration. Geographic and economic access barriers were identified in rural areas. Oral rehydration salts were provided as a part of household treatment in about 75% of children. 20% of them received non-medical care, specially from traditional healers. Primary medical care was provided to 60% of cases; the drawbacks of the process were related to a high rate of medication prescription, lack or referral to hospital, and lack of recommendation for a new visit in presence of alarm signs. Public physicians and those from rural communities prescribed oral rehydration therapy in a greater proportion. The problem of access to health services in rural communities was confirmed. However, low quality of care provided by primary level physicians was found in both rural and urban areas. We propose the creation of clinical teaching centers to improve the quality of the process of medical care as a strategy to decrease mortality due to AD.


Translational Regenerative Therapies for Chronic Spinal Cord Injury.
Spinal cord injury is a chronic and debilitating neurological condition that is currently being managed symptomatically with no real therapeutic strategies available. Even though there is no consensus on the best time to start interventions, the chronic phase is definitely the most stable target in order to determine whether a therapy can effectively restore neurological function. The advancements of nanoscience and stem cell technology, combined with the powerful, novel neuroimaging modalities that have arisen can now accelerate the path of promising novel therapeutic strategies from bench to bedside. Several types of stem cells have reached up to clinical trials phase II, including adult neural stem cells, human spinal cord stem cells, olfactory ensheathing cells, autologous Schwann cells, umbilical cord blood-derived mononuclear cells, adult mesenchymal cells, and autologous bone-marrow-derived stem cells. There also have been combinations of different molecular therapies; these have been either alone or combined with supportive scaffolds with nanostructures to facilitate favorable cell⁻material interactions. The results already show promise but it will take some coordinated actions in order to develop a proper step-by-step approach to solve impactful problems with neural repair.


Common Cell Shape Evolution of Two Nasopharyngeal Pathogens.
Respiratory infectious diseases are the third cause of worldwide death. The nasopharynx is the portal of entry and the ecological niche of many microorganisms, of which some are pathogenic to humans, such as Neisseria meningitidis and Moraxella catarrhalis. These microbes possess several surface structures that interact with the actors of the innate immune system. In our attempt to understand the past evolution of these bacteria and their adaption to the nasopharynx, we first studied differences in cell wall structure, one of the strongest immune-modulators. We were able to show that a modification of peptidoglycan (PG) composition (increased proportion of pentapeptides) and a cell shape change from rod to cocci had been selected for along the past evolution of N. meningitidis. Using genomic comparison across species, we correlated the emergence of the new cell shape (cocci) with the deletion, from the genome of N. meningitidis ancestor, of only one gene: yacF. Moreover, the reconstruction of this genetic deletion in a bacterium harboring the ancestral version of the locus together with the analysis of the PG structure, suggest that this gene is coordinating the transition from cell elongation to cell division. Accompanying the loss of yacF, the elongation machinery was also lost by several of the descendants leading to the change in the PG structure observed in N. meningitidis. Finally, the same evolution was observed for the ancestor of M. catarrhalis. This suggests a strong selection of these genetic events during the colonization of the nasopharynx. This selection may have been forced by the requirement of evolving permissive interaction with the immune system, the need to reduce the cellular surface exposed to immune attacks without reducing the intracellular storage capacity, or the necessity to better compete for adhesion to target cells.


Active site-directed double mutants of dihydrofolate reductase.
Variants of dihydrofolate reductase (DHFR), which confer resistance to antifolates, are used as dominant selectable markers in vitro and in vivo and may be useful in the context of gene therapy. To identify improved mutant human DHFRs with increased catalytic efficiency and decreased binding to methotrexate, we constructed by site-directed mutagenesis four variants with substitutions at both Leu22 and Phe31 (i.e., Phe22-Ser31, Tyr22-Ser31, Phe22-Gly31, and Tyr22-Gly31). Antifolate resistance has been observed previously when individual changes are made at these active-site residues. Substrate and antifolate binding properties of these "double" mutants revealed that each have greatly diminished affinity for antifolates (> 10,000-fold) yet only slightly reduced substrate affinity. Comparison of in vitro measured properties with those of single-residue variants indicates that double mutants are indeed significantly superior. This was verified for one of the double mutants that provided high-level methotrexate resistance following retrovirus-mediated gene transfer in NIH3T3 cells.


Recombinant Tissue Plasminogen Activator Induces Neurological Side Effects Independent on Thrombolysis in Mechanical Animal Models of Focal Cerebral Infarction: A Systematic Review and Meta-Analysis.
Recombinant tissue plasminogen activator (rtPA) is the only effective drug approved by US FDA to treat ischemic stroke, and it contains pleiotropic effects besides thrombolysis. We performed a meta-analysis to clarify effect of tissue plasminogen activator (tPA) on cerebral infarction besides its thrombolysis property in mechanical animal stroke. Relevant studies were identified by two reviewers after searching online databases, including Pubmed, Embase, and ScienceDirect, from 1979 to 2016. We identified 6, 65, 17, 12, 16, 12 and 13 comparisons reporting effect of endogenous tPA on infarction volume and effects of rtPA on infarction volume, blood-brain barrier, brain edema, intracerebral hemorrhage, neurological function and mortality rate in all 47 included studies. Standardized mean differences for continuous measures and risk ratio for dichotomous measures were calculated to assess the effects of endogenous tPA and rtPA on cerebral infarction in animals. The quality of included studies was assessed using the Stroke Therapy Academic Industry Roundtable score. Subgroup analysis, meta-regression and sensitivity analysis were performed to explore sources of heterogeneity. Funnel plot, Trim and Fill method and Egger's test were obtained to detect publication bias. We found that both endogenous tPA and rtPA had not enlarged infarction volume, or deteriorated neurological function. However, rtPA would disrupt blood-brain barrier, aggravate brain edema, induce intracerebral hemorrhage and increase mortality rate. This meta-analysis reveals rtPA can lead to neurological side effects besides thrombolysis in mechanical animal stroke, which may account for clinical exacerbation for stroke patients that do not achieve vascular recanalization with rtPA.


Generic pain intensity scores are affected by painful comorbidity.
To determine the degree to which the generic pain intensity rating (i.e., overall and without reference to a particular body site) of facial pain patients being seen in a specialty setting for facial pain is influenced by painful comorbidity in body parts other than the face. In this prospective study, 40 consecutive female temporomandibular pain patients rated their generic pain on a 100-mm visual analog scale. After marking all painful body sites on pain drawings, patients were asked to rate the pain intensity for each of the indicated pain sites; the patients did not have access to the generic pain intensity score. Pearson's correlation coefficient was used to correlate the generic pain intensity score with site-specific pain intensity ratings, their mean and maximum, and the number of pain sites. The medians of the generic, maximum, and facial pain intensity scores were 49.5, 53, and 45.5, respectively. The generic pain intensity rating correlated more highly with the intensity scores reported for the most painful body site (r2 = 0.82; P < 0.001) than with the average rating across all painful sites (r2 = 0.62; P < 0.001), or the pain intensity score in the face (r2 = 0.61; P < 0.001). The number of pain sites did not correlate to any statistically significant degree with the generic pain intensity rating (r2 = 0.006; P = 0.65). The results of this study suggest that the maximum visual analog scale pain intensity score, observed in any body location, is a better reflection of the generic pain intensity rating than the corresponding score of the face. To avoid over-rating or underrating of facial pain intensity, patients should be instructed to provide site-specific pain intensity scores if painful comorbidity is present.


A human model of epithelial to mesenchymal transition to monitor drug efficacy in hepatocellular carcinoma progression.
The epithelial to mesenchymal transition (EMT) of malignant hepatocytes is a crucial event in hepatocellular carcinoma (HCC) progression and recurrence. We aimed to establish a human model of EMT to examine drug efficacy and specificity in HCC progression. Human HCC cell populations were characterized by immunofluorescence analysis, migration and invasion assays, array comparative genomic hybridization, whole-genome expression profiling, and promoter methylation. Therapeutic agents clinically used against HCC were examined for efficacy by determination of IC(50) values. We show that liver cancer cell lines exhibited either an epithelial or mesenchymal phenotype of which the latter showed strong migratory and invasive abilities in vitro. The common cellular origin of both cell types indicated that mesenchymal HCC cells have been derived from epithelial hepatocytes through EMT in the HCC patient. Drug exposure of mesenchymal HCC cells showed higher resistance to the targeted therapeutic agents sorafenib and erlotinib as compared to epithelial HCC cells, which were slightly more resistant to cytostatic drugs. Most remarkably, combined treatment with doxorubicin and sorafenib caused increased susceptibility of both HCC cell types resulting in enhanced drug efficacy. Taken together, this EMT model of human HCC allows the identification of molecular mechanisms and the assessment of therapeutic drug efficacy during liver cancer progression in preclinical studies.


Non-Verbal Cognitive Abilities in Children and Adolescents Affected by Migraine and Tension-Type Headache: An Observational Study Using the Leiter-3.
Headache is one of the most common neurological disorders in developmental age. Several studies investigated the relationship between headache and emotional/behavioral problems. We studied non-verbal cognitive abilities, including non-verbal memory and attention skills, in order to evaluate the impact of primary headache on these domains. The latest version of the cognitive battery Leiter International Performance Scale - Third Edition (Leiter-3), a non-verbal test, was administered to 35 children and adolescents affected by migraine or tension-type headache and to 23 healthy subjects. We found that frequency of attacks and headache disability (evaluated with the Pediatric Migraine Disability Assessment Score Questionnaire) significantly correlate with non-verbal memory and sustained attention skills. However, we found that headache disability has a significant impact on specific cognitive domains related to sustained attention and non-verbal memory skills. The relationship between headache and memory/attention deficits may have an explanation based on a possible common physiopathology ground, including noradrenergic and dopaminergic pathways.


Vitamin D Level is Associated with Increased Left Ventricular Mass and Arterial Stiffness in Older Patients with Impaired Renal Function.
BACKGROUND Impaired renal function is common among older patients. Deficiency of vitamin D is a frequent phenomenon among patients with impaired renal function, who are likely to develop cardiovascular diseases. This study aimed to explore the association of 25 (OH) D levels with left ventricular mass and arterial stiffness in older patients with impaired renal function. MATERIAL AND METHODS Based on their admission estimate glomerular filtration rate (eGFR), 273 inpatients (≥65 years) were allocated into a normal eGFR group (≥60 ml/min) and an impaired eGFR group (<60 ml/min). The 25 (OH) D levels were measured and the left ventricular mass index (LVMI) was estimated. Pulse wave velocity (PWV) was used to explore arterial stiffness. RESULTS The 25 (OH) D levels of patients in the impaired eGFR group were significantly lower than in the normal eGFR group [(11.92±6.01) μg/L vs. (18.14±8.07) μg/L, p<0.05). LVMI and PWV were both significantly higher in the impaired eGFR group than in the normal eGFR group [(104.89±33.50) g/m2 vs. (92.95±18.95) g/m2, P<0.05; (15.99±3.10) m/s vs. (13.62±2.90) m/s, P<0.05]. After adjusting for age, sex, eGFR, cardiovascular risk factors, serum calcium, and iPTH levels, the inverse association between LVMI and 25 (OH) D, PWV, and 25 (OH) D were statistically significant. CONCLUSIONS Vitamin D level is lower in older patients with impaired renal function. Lower vitamin D levels were correlated with higher left ventricular mass and increased arterial stiffness in older patients.


Neural circuitry underlying sustained attention in healthy adolescents and in ADHD symptomatology.
Moment-to-moment reaction time variability on tasks of attention, often quantified by intra-individual response variability (IRV), provides a good indication of the degree to which an individual is vulnerable to lapses in sustained attention. Increased IRV is a hallmark of several disorders of attention, including Attention-Deficit/Hyperactivity Disorder (ADHD). Here, task-based fMRI was used to provide the first examination of how average brain activation and functional connectivity patterns in adolescents are related to individual differences in sustained attention as measured by IRV. We computed IRV in a large sample of adolescents (n = 758) across 'Go' trials of a Stop Signal Task (SST). A data-driven, multi-step analysis approach was used to identify networks associated with low IRV (i.e., good sustained attention) and high IRV (i.e., poorer sustained attention). Low IRV was associated with greater functional segregation (i.e., stronger negative connectivity) amongst an array of brain networks, particularly between cerebellum and motor, cerebellum and prefrontal, and occipital and motor networks. In contrast, high IRV was associated with stronger positive connectivity within the motor network bilaterally and between motor and parietal, prefrontal, and limbic networks. Consistent with these observations, a separate sample of adolescents exhibiting elevated ADHD symptoms had increased fMRI activation and stronger positive connectivity within the same motor network denoting poorer sustained attention, compared to a matched asymptomatic control sample. With respect to the functional connectivity signature of low IRV, there were no statistically significant differences in networks denoting good sustained attention between the ADHD symptom group and asymptomatic control group. We propose that sustained attentional processes are facilitated by an array of neural networks working together, and provide an empirical account of how the functional role of the cerebellum extends to cognition in adolescents. This work highlights the involvement of motor cortex in the integrity of sustained attention, and suggests that atypically strong connectivity within motor networks characterizes poor attentional capacity in both typically developing and ADHD symptomatic adolescents.


Dextrose boluses versus burette dextrose infusions in prevention of hypoglycemia among preterms admitted at Mulago Hospital: an open label randomized clinical trial.
Hypoglycemia is a major cause of morbidity and mortality among preterm infants and its management remains a challenge in resource limited settings. Use of dextrose infusion by the recommended infusion pumps is not feasible in our environment due to their high costs and yet the current use of mini dextrose boluses with syringes as adapted at Mulago national referral and tertiary teaching hospital has unknown efficacy in prevention of hypoglycemia. We determined the efficacy of dextrose infusions by burettes versus two hourly dextrose boluses in prevention of hypoglycemia among preterms admitted in the first 72 hours at Special Care Unit, Mulago Hospital. One hundred and forty preterms aged 0 to 24 hours of life were randomized to receive 10% IV dextrose either as mini boluses or by infusion using burettes in an open label clinical trial. Blood glucose was measured at 0, two hourly for next 6 hours, 6 hourly for next 12 hours and thereafter 12 hourly until end of 72 hours following admission. Primary end point was incidence of hypoglycemia (random blood sugar (RBS) < 2.6 mmol/l) which was expressed as relative risk (RR). Efficacy of the dextrose infusion was computed using 1-RR. From February 2012 to April 2012, 68 preterms in the bolus arm and 72 in the infusion arm were studied. Hypoglycemia was detected in 34% (48/140). The incidence of hypoglycemia in the bolus arm was 59% (40/68) compared to 11% (8/72) in the infusion arm (RR; 0.19, 95% CI; 0.09-0.37). Efficacy (1-RR) of infusion by burettes versus boluses in prevention of hypoglycemia among preterms was 0.81 (95% CI; 0.63-0.90). Continuous 10% dextrose infusion by burettes reduced the incidence of hypoglycemia by 81% in the first 72 hours of admission compared to two hourly 10% mini dextrose boluses among preterms admitted at Special Care Unit, Mulago Hospital. (ClinicalTrials.gov Identifier: NCT01688674).


Genetic regulatory networks programming hematopoietic stem cells and erythroid lineage specification.
Erythroid cell production results from passage through cellular hierarchies dependent on differential gene expression under the control of transcription factors responsive to changing niches. We have constructed Genetic Regulatory Networks (GRNs) describing this process, based predominantly on mouse data. Regulatory network motifs identified in E. coli and yeast GRNs are found in combination in these GRNs. Feed-forward motifs with autoregulation generate forward momentum and also control its rate, which is at its lowest in hematopoietic stem cells (HSCs). The simultaneous requirement for multiple regulators in multi-input motifs (MIMs) provides tight control over expression of target genes. Combinations of MIMs, exemplified by the SCL/LMO2 complexes, which have variable content and binding sites, explain how individual regulators can have different targets in HSCs and erythroid cells and possibly also how HSCs maintain stem cell functions while expressing lineage-affiliated genes at low level, so-called multi-lineage priming. MIMs combined with cross-antagonism describe the relationship between PU.1 and GATA-1 and between two of their target genes, Fli-1 and EKLF, with victory for GATA-1 and EKLF leading to erythroid lineage specification. These GRNs are useful repositories for current regulatory information, are accessible in interactive form via the internet, enable the consequences of perturbation to be predicted, and can act as seed networks to organize the rapidly accumulating microarray data.


Measuring family functioning in families with parental cancer: Reliability and validity of the German adaptation of the Family Assessment Device (FAD).
The concept of family functioning is gaining importance in psycho-oncology research and health care services. The Family Assessment Device (FAD) is a well-established measure of family functioning. Psychometric properties inherent in the German 51-item adaptation of the FAD are examined in different samples of families with parental cancer. Acceptance, reliability, and validity of FAD scales are analysed in samples from different study settings (N=1701 cancer patients, N=261 partners, N=158 dependent adolescent children 11 to 18years old). Missing items in the FAD scales (acceptance) are rare for adults (<1.1%) and adolescent children (<4.4%). In samples of adults and older adolescents (15 to 18years), all FAD scales except for the Roles scale are significantly reliable (0.75≤Cronbach's α≤0.88). The scales correlate highly (0.46≤Pearson's r≤0.59) with the criterion satisfaction with family life (convergent validity), and have smaller correlations (0.16≤r≤0.49) with measures of emotional distress and subjective well-being (divergent validity). In most FAD scales, adults seeking family counselling report worse family functioning (0.24≤Cohen's d≤0.59) than adults in other samples with parental cancer (discriminative validity). Overall, the German 51-item adaptation of the FAD reveals good acceptance, reliability, and validity for cancer patients and their relatives. Particularly the scale General Functioning shows excellent psychometric properties. The FAD is suitable in the assessment of families with parental cancer for adults and adolescents older than 11years.


Episodic memory reconsolidation: updating or source confusion?
Reactivation of apparently stable, long-term memory can render it fragile, and dependent on a re-stabilisation process referred to as "reconsolidation". Recently we provided the first demonstration of reconsolidation effects in human episodic memory (Hupbach, Gomez, Hardt, & Nadel, 2007; Hupbach, Hardt, Gomez, & Nadel, 2008). Memory for a set of objects was modified by the presentation of a new set, if and only if participants were reminded of the first learning episode before learning the new set. The present study asks whether this effect can be interpreted as a source discrimination problem; i.e., participants have difficulties remembering which objects were presented during which session, and do not actually incorporate new objects into the reactivated memory. The present study used a recognition test and asked participants directly about the source of their memories. Participants in the no-reminder group showed very few source errors. Participants in the reminder group misattributed the source of objects from the second set as being from the first set but not vice versa, thus demonstrating updating of the original memory. This finding is informative with respect to the misinformation paradigm, and reconsolidation is discussed as a possible mechanism underlying our results and the misinformation effect.


Resulting Shifts in Percentile and Standard Placements after Comparison of the BOD POD and DXA.
The purpose of this study was to determine the validity of the BOD POD® when compared to the DXA and if placement on a percentile chart and standard table is affected by any differences between the two measures. A total of 244 (27.7 ± 10.8 yrs, 77.3 ± 16.1 kg, 171.4 ± 10.1 cm, 26.31 ± 5.42 BMI) males and females between the ages of 18 and 52 were recruited to participate in this study. The participant's body fat percentage (%BF) was tested in random order on the BOD POD® and DXA during a 30-minute session following manufacturer's guidelines and procedures. Dependent t-test indicated the %BF measured by the BOD POD® (23.4% ± 12.8) was significantly lower when compared to the DXA (29.5% ± 12.1), p = .001. The Pearson's Product moment correlation was 0.95 (p = .001), indicating a very strong relationship between the two instruments. Using estimates of %BF from the BOD POD® also resulted in more favorable shifts on a percentile chart and standard table. Since a high correlation was evident between the two, the BOD POD® can be used as an instrument to track %BF changes over time during a diet and/or exercise intervention. However, caution should be made when classifying %BF with percentile charts or standard tables using the BOD POD® %BF estimates.


Immune responses following neonatal DNA vaccination are long-lived, abundant, and qualitatively similar to those induced by conventional immunization.
Virus infections are devastating to neonates, and the induction of active antiviral immunity in this age group is an important goal. Here, we show that a single neonatal DNA vaccination induces cellular and humoral immune responses which are maintained for a significant part of the animal's life span. We employ a sensitive technique which permits the first demonstration and quantitation, directly ex vivo, of virus-specific CD8(+) T cells induced by DNA immunization. One year postvaccination, antigen-specific CD8(+) T cells were readily detectable and constituted 0.5 to 1% of all CD8(+) T cells. By several criteria-including cytokine production, perforin content, development of lytic ability, and protective capacity-DNA vaccine-induced CD8(+) memory T cells were indistinguishable from memory cells induced by immunization with a conventional (live-virus) vaccine. Analyses of long-term humoral immune responses revealed that, in contrast to the strong immunoglobulin G2a (IgG2a) skewing of the humoral response seen after conventional vaccination, IgG1 and IgG2a levels were similar in DNA-vaccinated neonatal and adult animals, indicating a balanced T helper response. Collectively, these results show that a single DNA vaccination within hours or days of birth can induce long-lasting CD8(+) T- and B-cell responses; there is no need for secondary immunization (boosting). Furthermore, the observed immune responses induced in neonates and in adults are indistinguishable by several criteria, including protection against virus challenge.


Influence of fatigue on hand muscle coordination and EMG-EMG coherence during three-digit grasping.
Fingertip force control requires fine coordination of multiple hand muscles within and across the digits. While the modulation of neural drive to hand muscles as a function of force has been extensively studied, much less is known about the effects of fatigue on the coordination of simultaneously active hand muscles. We asked eight subjects to perform a fatiguing contraction by gripping a manipulandum with thumb, index, and middle fingers while matching an isometric target force (40% maximal voluntary force) for as long as possible. The coordination of 12 hand muscles was quantified as electromyographic (EMG) muscle activation pattern (MAP) vector and EMG-EMG coherence. We hypothesized that muscle fatigue would cause uniform changes in EMG amplitude across all muscles and an increase in EMG-EMG coherence in the higher frequency bands but with an invariant heterogeneous distribution across muscles. Muscle fatigue caused a 12.5% drop in the maximum voluntary contraction force (P < 0.05) at task failure and an increase in the SD of force (P < 0.01). Although EMG amplitude of all muscles increased during the fatiguing contraction (P < 0.001), the MAP vector orientation did not change, indicating that a similar muscle coordination pattern was used throughout the fatiguing contraction. Last, EMG-EMG coherence (0-35 Hz) was significantly greater at the end than at the beginning of the fatiguing contraction (P < 0.01) but was heterogeneously distributed across hand muscles. These findings suggest that similar mechanisms are involved for modulating and sustaining digit forces in nonfatiguing and fatiguing contractions, respectively.


Fertility and infertility: Definition and epidemiology.
Infertility is a disease characterized by the failure to establish a clinical pregnancy after 12 months of regular and unprotected sexual intercourse. It is estimated to affect between 8 and 12% of reproductive-aged couples worldwide. Males are found to be solely responsible for 20-30% of infertility cases but contribute to 50% of cases overall. Secondary infertility is the most common form of female infertility around the globe, often due to reproductive tract infections. The three major factors influencing the spontaneous probability of conception are the time of unwanted non-conception, the age of the female partner and the disease-related infertility. The chance of becoming spontaneously pregnant declines with the duration before conception. The fertility decline in female already starts around 25-30 years of age and the median age at last birth is 40-41 years in most studied populations experiencing natural fertility. The disease-related infertility may affect both genders or be specific to one gender. The factors affecting both genders' fertility are hypogonadotrophic hypogonadism, hyperprolactinemia, disorders of ciliary function, cystic fibrosis, infections, systemic diseases and lifestyle related factors/diseases. Premature ovarian insufficiency, polycystic ovary syndrome, endometriosis, uterine fibroids and endometrial polyps may play a role in female infertility. Male infertility may be due to testicular and post-testicular deficiencies. Semen decline that has been observed over the years, endocrine disrupting chemicals and consanguinity are other factors that may be involved.


Comparative analysis of telmisartan and olmesartan on cardiac function in the transgenic (mRen2)27 rat.
Telmisartan, an angiotensin receptor blocker, may have unique benefits as it possesses partial peroxisome proliferator-activated receptor (PPAR)-γ agonist activity in addition to antihypertensive effects. In this study, we test whether treatment with telmisartan ameliorates cardiovascular abnormalities to a greater extent than olmesartan, which has little PPAR-γ activity. The hypertensive rodent model of tissue renin-angiotensin system activation, transgenic (mRen2)27 (Ren2) rats and their littermate Sprague-Dawley controls were used. Rats were treated with telmisartan (2 mg · kg(-1) · day(-1)), olmesartan (2.5 mg · kg(-1) · day(-1)), or vehicle via drinking water for 3 wk; these doses achieved similar blood pressure control, as measured by telemetry. Ren2 rats displayed impaired diastolic and systolic function using left ventricular (LV) pressure-volume (P-V) analysis. Load-independent diastolic indexes, including the time constant of isovolumic relaxation and the slope of the end-diastolic P-V relationship, as well as systolic indexes, including preload recruitable stroke work, the dP/dt(max)-end-diastolic volume (EDV) relationship, and the P-V area-EDV relationship, were elevated in Ren2 rats compared with Sprague-Dawley controls (P < 0.05). The Ren2 myocardium exhibited parallel increases in the oxidant markers NADPH oxidase and 3-nitrotyrosine. The increase in the prohypertrophic protein Jak2 in Ren2 rats was associated with cardiac structural abnormalities using light microscopic and ultrastructural analysis, which included interstitial fibrosis, cardiomyocyte and LV hypertrophy, and mitochondrial derangements. Both angiotensin receptor blockers attenuate these abnormalities to a similar extent. Our data suggest that the beneficial effect of telmisartan and olmesartan on cardiac structure and function may be predominantly pressor-related or angiotensin type 1 receptor dependent in this model of renin-angiotensin system activation.


Preparative Separation of Three Monoterpenes from Perilla frutescens var. crispa Using Centrifugal Partition Chromatography.
Three monoterpenes, namely, 9-hydroxy isoegomaketone (1), isoegomaketone (2), and perilla ketone (3), were successfully separated from the supercritical carbon dioxide (SC-CO2) extract of the leaves of Perilla frutescens var. crispa (cv. Antisperill; Lamiaceae) by centrifugal partition chromatography (CPC). To obtain large quantities of these materials required for studies on their mechanism of action and in vivo effectiveness in inflammation, we used CPC because of its high loading capacity and reproducibility to purify the three compounds. Compound 1 (2.60 mg, 96.7% purity at 254 nm) was purified from 500 mg of the SC-CO2 extract of P. frutescens var. crispa (cv. Antisperill), using a two-phase solvent system comprising n-hexane/ethyl acetate/ethanol/water (5:5:5:5 v/v) in a descending mode. As compounds 2 (56.1 mg, 97.6% purity at 254 nm) and 3 (78.6 mg, 96.1% purity at 254 nm) are highly volatile and difficult to recover from an aqueous mobile phase after purification during the drying process, they were obtained from the same amount of the processed extract in an ascending mode using the upper organic phase as the mobile phase (n-hexane/ethyl acetate/ethanol/water, 8:2:8:2 v/v). The structures of compounds 1-3 were confirmed by 1H- and 13C-NMR analysis. Thus, based on our findings, we recommend centrifugal partition chromatography as a powerful technique for purifying the active principal compounds 1 and 2 from the leaves of P. frutescens var. crispa.


Assessment of the accuracy of lacrimal scintigraphy based on a prospective analysis of patients' symptomatology.
Dacryoscintigraphy is a noninvasive method frequently used for assessment of the lacrimal drainage system. Sometimes conflicting results with patients' complaints are obtained, which have no clear explanation. In our hypothesis, follow-up of patients evaluated with dacryoscintigraphy could be helpful in determining possible explanations for these conflicting results. Thirty-eight consecutive patients (76 eyes) who had not previously undergone dacryocystorhinostomy or probing were entered in the study. Following instillation of 4 MBq (99m)Tc-pertechnetate, a routine procedure of dacryoscintigraphy was performed. After a mean follow-up time of 1.2 +/- 0.3 years (range 0.9-1.6 years), all patients were reassessed clinically. Using initial patients' symptoms as the gold standard, the sensitivity, specificity, NPV, and PPV of dacryoscintigraphy were calculated to be 82.1%, 75%, 60%, and 90.1%, respectively. The kappa value as a measure of agreement was 0.52. Considering the follow-up study as the gold standard, the sensitivity, specificity, NPV, and PPV were calculated as follows, respectively: 96.3%, 90.4%, 96.3%, and 90.4%. The kappa value was 0.86. There were three initially symptom-free eyes with obstructive pattern on the scans, which developed epiphora on the follow-up. In fact, these eyes were initially classified as false positive, but follow-up assessment showed that they were true-positive scans in a subclinical state. Despite certain drawbacks inherent to the technique, dacryoscintigraphy provides valuable independent information and allows more accurate management decisions. In some patients, data obtained from lacrimal scintigraphy could be predictive, and it makes possible determination of subclinical nasolacrimal duct obstructions.


Characterization of Ammonia, Methane, and Nitrous Oxide Emissions from Concentrated Animal Feeding Operations in Northeastern Colorado.
Atmospheric emissions from animal husbandry are important to both air quality and climate, but are hard to characterize and quantify as they differ significantly due to management practices and livestock type, and they can vary substantially throughout diurnal and seasonal cycles. Using a new mobile laboratory, ammonia (NH3), methane (CH4), nitrous oxide (N2O), and other trace gas emissions were measured from four concentrated animal feeding operations (CAFOs) in northeastern Colorado. Two dairies, a beef cattle feedlot, and a sheep feedlot were chosen for repeated diurnal and seasonal measurements. A consistent diurnal pattern in the NH3 to CH4 enhancement ratio is clearly observed, with midday enhancement ratios approximately four times greater than nighttime values. This diurnal pattern is similar, with slight variations in magnitude, at the four CAFOs and across seasons. The average NH3 to CH4 enhancement ratio from all seasons and CAFOs studied is 0.17 (+0.13/-0.08) mol/mol, in agreement with statewide inventory averages and previous literature. Enhancement ratios for NH3 to N2O and N2O to CH4 are also reported. The enhancement ratios can be used as a source signature to distinguish feedlot emissions from other NH3 and CH4 sources, such as fertilizer application and fossil fuel development, and the large diurnal variability is important for refining inventories, models, and emission estimates.


Mediterranean Diet, Its Components, and Amyloid Imaging Biomarkers.
There is accumulating evidence suggesting that diet may play a role in preventing or delaying cognitive decline and dementia, but the underlying biological mechanisms are not well understood. To examine the cross-sectional associations of the Mediterranean diet (MeDi) and its components with 11C-PiB-PET scan measures of amyloid-β (Aβ) deposition. The study consisted of 278 Mayo Clinic Study of Aging participants 70+ years old, who were cognitively unimpaired (CU) at the time of completion of the Food Frequency Questionnaire (FFQ) and when they underwent PET imaging. Adherence to the MeDi was assessed by computing the MeDi score for each participant. All scans were performed after the FFQ completion; median [IQR] time between FFQ and Aβ PET was 3.5 (1.4) years. Z-scores were created for component, macro- and micronutrients measured. Linear and logistic regression models were adjusted for age, sex, education, apolipoprotein E (APOE) ɛ4 allele carrier status, time interval between the FFQ completion and PET scan, and total energy intake. Participants' median age at FFQ was 77.7 years (55.8% men; 26.6% with an APOE ɛ4 allele). Higher MeDi score (linear regression slope (beta):-0.035, p = 0.012; per standard deviation increase), vegetable intake (beta:-0.043, p = 0.002), intake of vitamin A (beta:-0.041, p = 0.003) or β-carotene (beta: -0.039, p = 0.005) from food sources and moderate alcohol consumption (beta: -0.074, p = 0.03) were associated with lower 11C-PiB standardized uptake value ratio. Findings are consistent with previous studies suggesting that higher adherence to a MeDi pattern and higher vegetable consumption are associated with better neuroimaging biomarker profile. Prospective studies are needed to validate current findings.


A technical report on repair of amalgam-dentin complex.
This clinical report describes a repair protocol for cusp fracture of a failed amalgam-dentin complex. A maxillary right first premolar with an amalgam restoration presented a buccal cusp fracture. Chairside repair has been undertaken by conditioning the existing amalgam restoration with silica coating (30 μm CoJet®-Sand), phosphoric acid etching the beveled enamel surface, priming dentin, and application of a bonding agent on both enamel and dentin. Thereafter, the amalgam was silanized (ESPE®-Sil), and opaque resin was applied and polymerized to mask the amalgam. The fractured buccal cusp was modeled using resin composite (Clearfil Photo Posterior) and photo-polymerized. Finally, the amalgam was refinished and refurbished and the composite was finished and polished.


Small-scale indirect plant responses to insect herbivory could have major impacts on canopy photosynthesis and isoprene emission.
Insect herbivores cause substantial changes in the leaves they attack, but their effects on the ecophysiology of neighbouring, nondamaged leaves have never been quantified in natural canopies. We studied how winter moth (Operophtera brumata), a common herbivore in temperate forests, affects the photosynthetic and isoprene emission rates of its host plant, the pedunculate oak (Quercus robur). Through a manipulative experiment, we measured leaves on shoots damaged by caterpillars or mechanically by cutting, or left completely intact. To quantify the effects at the canopy scale, we surveyed the extent and patterns of leaf area loss in the canopy. Herbivory reduced photosynthesis both in damaged leaves and in their intact neighbours. Isoprene emission rates significantly increased after mechanical leaf damage. When scaled up to canopy-level, herbivory reduced photosynthesis by 48 ± 10%. The indirect effects of herbivory on photosynthesis in undamaged leaves (40%) were much more important than the direct effects of leaf area loss (6%). If widespread across other plant-herbivore systems, these findings suggest that insect herbivory has major and previously underappreciated influences in modifying ecosystem carbon cycling, with potential effects on atmospheric chemistry.


Pyruvate decarboxylase provides growing pollen tubes with a competitive advantage in petunia.
Rapid pollen tube growth places unique demands on energy production and biosynthetic capacity. The aim of this work is to understand how primary metabolism meets the demands of such rapid growth. Aerobically grown pollen produce ethanol in large quantities. The ethanolic fermentation pathway consists of two committed enzymes: pyruvate decarboxylase (PDC) and alcohol dehydrogenase (ADH). Because adh mutations do not affect male gametophyte function, the obvious question is why pollen synthesize an abundant enzyme if they could do just as well without. Using transposon tagging in Petunia hybrida, we isolated a null mutant in pollen-specific Pdc2. Growth of the mutant pollen tubes through the style is reduced, and the mutant allele shows reduced transmission through the male, when in competition with wild-type pollen. We propose that not ADH but rather PDC is the critical enzyme in a novel, pollen-specific pathway. This pathway serves to bypass pyruvate dehydrogenase enzymes and thereby maintain biosynthetic capacity and energy production under the unique conditions prevailing during pollen-pistil interaction.


Memory, anticipation, action - working with Troy D. Zars.
We present here our reflections on the scientific work of the late Troy D. Zars (1967 - 2018), on what it was like to work with him, and what it means to us. A common theme running through his work is that memory systems are not for replaying the past. Rather, they are forward-looking systems, providing whatever guidance past experience has to offer for anticipating the outcome of future actions. And in situations where no such guidance is available trying things out is the best option. Working with Troy was inspiring precisely because of the optimism inherent in this concept and that he himself embodied. Our reflections highlight what this means to us as his former mentors, colleagues, and mentees, respectively, and what it might mean for the future of neurogenetics.


Validation of the swimming competence questionnaire for children.
Two studies were employed to test the reliability and validity of the Swimming Competence Questionnaire (SCQ) among primary school children. Study 1 was a cross-sectional survey in 4959 primary school children. Study 2 was a pre-post-test quasi-experiment among 1609 primary school children who underwent a 20-lesson learn-to-swim programme. In Study 1, exploratory structural equation modelling revealed excellent goodness-of-fit and scale reliability for a two-factor model comprising distance and skill factors, which supported the construct and convergent validity. SCQ scores were significantly and positively correlated with swimming outcomes (i.e., self-efficacy, intention, swimming frequency), which supported SCQ's concurrent and criterion validity. Average variance extracted for the SCQ factors exceeded cut-off criteria supporting discriminant validity. In Study 2, pre-test SCQ scores correlated significantly and positively with the SCQ scores, self-efficacy, intention, and swimming frequency at post-test, which supported SCQ's test-retest reliability and predictive validity. Positive intraclass correlation between SCQ scores and coach ratings at post-test provided evidence for SCQ's inter-rater reliability. SCQ scores significantly improved at post-test, which supported SCQ's ecological validity. In conclusion, findings indicate that the SCQ is a valid and reliable measure to assess primary school children's swimming competence, in terms of swimming distance and basic water survival skills.


Is Streptococcus pyogenes resistant or susceptible to trimethoprim-sulfamethoxazole?
Streptococcus pyogenes is commonly believed to be resistant to trimethoprim-sulfamethoxazole (SXT), resulting in reservations about using SXT for skin and soft tissue infections (SSTI) where S. pyogenes is involved. S. pyogenes' in vitro susceptibility to SXT depends on the medium's thymidine content. Thymidine allows S. pyogenes to bypass the sulfur-mediated inhibition of folate metabolism and, historically, has resulted in apparently reduced susceptibility of S. pyogenes to sulfur antibacterials. The low thymidine concentration in Mueller-Hinton agar (MHA) is now regulated. We explored S. pyogenes susceptibility to SXT on various media. Using two sets of 100 clinical S. pyogenes isolates, we tested for susceptibility using SXT Etests on MHA containing defibrinated horse blood and 20 mg/liter β-NAD (MHF), MHA with sheep blood (MHS), MHA alone, MHA with horse blood (MHBA), and MHA with lysed horse blood (MHLHBA). European Committee on Antibacterial Susceptibility Testing (EUCAST) breakpoints defined susceptibility (MIC, ≤ 1 mg/liter) and resistance (MIC, >2 mg/liter). In study 1, 99% of S. pyogenes isolates were susceptible to SXT on MHA, MHBA, and MHLHBA, with geometric mean MICs of 0.04, 0.04, and 0.05 mg/liter, respectively. In study 2, all 100 S. pyogenes isolates were susceptible to SXT on MHF, MHS, MHA, and MHLHBA with geometric mean MICs of 0.07, 0.16, 0.07, and 0.09 mg/liter, respectively. This study confirms the in vitro susceptibility of S. pyogenes to SXT, providing support for the use of SXT for SSTIs. A clinical trial using SXT for impetigo is ongoing.


Sensitivity of locus ceruleus neurons to reward value for goal-directed actions.
The noradrenergic nucleus locus ceruleus (LC) is associated classically with arousal and attention. Recent data suggest that it might also play a role in motivation. To study how LC neuronal responses are related to motivational intensity, we recorded 121 single neurons from two monkeys while reward size (one, two, or four drops) and the manner of obtaining reward (passive vs active) were both manipulated. The monkeys received reward under three conditions: (1) releasing a bar when a visual target changed color; (2) passively holding a bar; or (3) touching and releasing a bar. In the first two conditions, a visual cue indicated the size of the upcoming reward, and, in the third, the reward was constant through each block of 25 trials. Performance levels and lipping intensity (an appetitive behavior) both showed that the monkeys' motivation in the task was related to the predicted reward size. In conditions 1 and 2, LC neurons were activated phasically in relation to cue onset, and this activation strengthened with increasing expected reward size. In conditions 1 and 3, LC neurons were activated before the bar-release action, and the activation weakened with increasing expected reward size but only in task 1. These effects evolved as monkeys progressed through behavioral sessions, because increasing fatigue and satiety presumably progressively decreased the value of the upcoming reward. These data indicate that LC neurons integrate motivationally relevant information: both external cues and internal drives. The LC might provide the impetus to act when the predicted outcome value is low.


The Effectiveness of Community-Based Nutrition Education on the Nutrition Status of Under-five Children in Developing Countries. A Systematic Review.
This systematic review aimed at examining the best available evidence on the effectiveness of community-based nutrition education in improving the nutrition status of under five children in developing countries. A systematic search of the literature was conducted utilising the following data bases: Cumulative Index to Nursing and Allied Health Literature (CINAHL), EMBASE, Medline, and Web of Knowledge. 9 studies were identified for the critical appraisal process. The Joanna Briggs Institute (JBI) critical appraisal check-list for experimental studies was utilised and two reviewers conducted the appraisal process independently. 7 studies were included for this review and data was extracted using the JBI data extraction form for experimental studies. The extracted data was heterogeneous as such narrative synthesis was conducted. The nutritional status of children in all studies improved and this was evidenced by increases in weight, height, mid upper arm circumference and reduced morbidity. Key messages about education were age at introduction of complementary foods, nutrition value on different types of feeds found locally and frequency of feeding the children. However, there were varied results regarding the effects of the intervention on the nutrition status of children. This was attributed by differences in implementers' characteristics, different intervention strategy and intensity, difference in age of the children at enrolment, pre-existing children's growth and nutritional status and follow-up periods. In addition to home visiting, conducting group meetings of care givers and community leaders, providing education twice a week and use of cooking demonstrations have shown that they produce highly significant findings. The evidence from the identified studies suggests that community- based nutrition education improves the nutrition status of under-five children in developing countries.


Interactions among white spruce tannins, Bacillus thuringiensis subsp. kurstaki, and spruce budworm (Lepidoptera: Tortricidae), on larval survival, growth, and development.
The interactions among white spruce, Picea glauca (Moench) Voss, purified acetone tannin extracts (hydrolyzable and condensed tannin), Bacillus thuringiensis subsp. kurstaki Cry1A(c) delta-endotoxin strain HD-73 (Btk), and spruce budworm, Choristoneura fumiferana (Clemens) (Lepidoptera: Tortricidae) on larval survival, growth, and development were investigated over the whole larval feeding period by using artificial diet supplemented with three concentrations of Btk toxins per milliliter of diet (0, 0.021, and 1.72 microg/ml) and three concentrations of foliar tannin extract (0, 8, and 15% dry mass basis). At high Btk concentration, tannin antagonized Btk potency against spruce budworm by lowering Btk-related larval mortality from 83 to 43%. At moderate Btk concentration tannin did not affect Btk potency. Host tree tannins antagonized not only the lethal effects of Btk toxin but also sublethal Btk-related impacts in terms of larval development, pupal weight, relative consumption rate, and growth rate. When alone in the diet, tannin negatively affected larval survival, growth, and development. Maximum potency of tannins against spruce budworm larvae (60% mortality) was reached at dietary concentrations corresponding to what is found in the plant (8% dry mass). The addition of Btk toxin in food containing tannin reduced percentage of larval mortality by one-third, indicating that Btk toxin can antagonize tannin potency against the insect. Development of Btk transgenic spruce trees should consider the antagonistic effect the toxin may have on the resistance conferred by tannins that have evolved naturally in spruce trees.


Previewing distracters reduces their effective contrast.
In a visual search task, when half the distracters are presented earlier than the remainder ('previewed'), observers find the target item more efficiently than when all the items are presented together -- the preview benefit. We measured psychometric functions for contrast increments on Gabors that were presented as a valid preview for subsequent search, and when they were a non-predictive (dummy) preview. Sensitivity to contrast increments was lower (rightwards shift of the psychometric function) on valid, compared to dummy previews. This is consistent with an account of the preview benefit in terms of active inhibition, equivalent to lowering the contrast of previewed items that are being actively ignored.


Red blood cell changes during the evolution of the sickle cell painful crisis.
A longitudinal study of the red blood cell (RBC) deformability, percent of dense erythrocytes, and hematologic parameters has been conducted during 117 painful crises affecting 36 patients with sickle cell anemia between January, 1985 and December, 1990. RBC deformability was determined by osmotic gradient ektacytometry and the percentage of dense cells was quantitated by centrifugation on a discontinuous Stractan density gradient. The data indicate that the painful crisis is a process that follows a bimodal form of evolution. The first phase of the painful crisis is characterized by increase in the severity of pain, increase in the number of dense cells, and a decrease in RBC deformability. In some patients the changes in dense cells and RBC deformability are evident 1 to 3 days before the onset of pain. In addition, the hemoglobin level decreases and the reticulocyte count increases during this initial phase. The second phase of the crisis is characterized by reduction in pain intensity, decrease in the number of dense cells, and increase in RBC deformability to values higher than those seen in the steady state. Moreover, the improvement in RBC deformability and the decrease in the number of dense cells at the end of a crisis seem to constitute new risk factors that may incite a recurrence of the crisis within 1 month in about 50% of painful episodes. The pathophysiologic events responsible for this bimodal behavior of RBCs during painful episodes may represent the appearance of factors that induce (1) preferential trapping of deformable cells in the microcirculation during the first phase of the crisis, followed by a decrease of dense cells and the appearance of new deformable RBCs released from the bone marrow during the second phase of the crisis; or (2) variable sickling of all circulating RBCs during the first phase followed by disappearance of dense RBCs and their replenishment by deformable cells during the second phase.


Too materialistic to get married and have children?
We developed new materials to induce a luxury mindset and activate materialistic values, and examined materialism's relationship to attitudes toward marriage and having children in Singapore. Path analyses indicated that materialistic values led to more negative attitudes toward marriage, which led to more negative attitudes toward children, which in turn led to a decreased number of children desired. Results across two studies highlight, at the individual level, the tradeoff between materialistic values and attitudes toward marriage and procreation and suggest that a consideration of psychological variables such as materialistic values may allow for a better understanding of larger-scale socioeconomic issues including low fertility rates among developed countries. We discuss implications and describe how psychological factors relating to low fertility fit within evolutionary mismatch and life history theory frameworks.


A peptide-based, ratiometric biosensor construct for direct fluorescence detection of a protein analyte.
We present the design, synthesis, and functional evaluation of peptide-based fluorescent constructs for wavelength-ratiometric biosensing of a protein analyte. The concept was shown using the high-affinity model interaction between the 18 amino acid peptide pTMVP and a recombinant antibody fragment, Fab57P. pTMVP was functionalized in two different positions with 6-bromomethyl-2-(2-furanyl)-3-hydroxychromone, an environmentally sensitive fluorophore with a two-band emission. The equilibrium dissociation constant of the interaction between pTMVP and Fab57P was largely preserved upon labeling. The biosensor ability of the labeled peptide constructs was evaluated in terms of the relative intensity change of the emission bands from the normal (N*) and tautomer (T*) excited-state species of the fluorophore ( I(N*)/I(T*)) upon binding of Fab57P. When the peptide was labeled in the C terminus, the I(N*)/I(T*) ratio changed by 40% upon analyte binding, while labeling close to the residues most important for binding resulted in a construct that completely lacked ratiometric biosensor ability. Integrated biosensor elements for reagentless detection, where peptides and ratiometric fluorophores are combined to ensure robustness in both recognition and signaling, are expected to become an important contribution to the design of future protein quantification assays in immobilized formats.


Time trends in ophthalmia neonatorum and dacryocystitis of the newborn in England, 2000-2011: database study.
To investigate English national trends in ophthalmia neonatorum and dacryocystitis (ON) of the newborn and the completeness of statutory notification of this serious infection. Analysis of hospital episode statistics (HES) from 2000 to 2011. Using linked HES, numbers of neonates hospitalised with ON were identified from 2000 to 2011. The numbers of hospitalised cases were compared with numbers of statutory notifications for ON published by the Notifications of Infectious Diseases (NOIDS). The national incidence rate of hospitalised cases showed a gradual decline from 464 (95% CI 447 to 482) per 100,000 live births in 2000 to 216 (204 to 228) per 100,000 live births in 2005. It then gradually increased to 471 (455 to 487) per 100,000 live births in 2010, but dropped to 257 (245 to 269) per 100,000 in 2011. From 2000 to 2009, when NOIDS data were available, the NOIDS data showed only 1006 cases compared with 20,505 cases in HES, and thus the notification system captured only about 1 case in 20. As shown by hospital statistics, there were marked cyclical fluctuations in ON over the study period. The annual figures for ON reported during the study period, under statutory health protection regulations, underestimated the actual occurrence of this disease by a very substantial amount. Linked hospital data should be used routinely to monitor the national incidence of ON.