Exploring the Effects of the Narrative Embodied in the Hearing Aid Fitting Process on Treatment Outcomes.
There is strong evidence from other fields of health, and growing evidence in audiology, that characteristics of the process of intervention as perceived by the client (embodied narratives) can have significant effects on treatment outcomes, independent of the technical properties of the intervention itself. This phenomenon deserves examination because studies of technical interventions that fail to take account of it may reach erroneous conclusions and because clinical practice can put such effects to therapeutic use. The aim of this study was to test the idea that embodied narratives might affect outcomes in hearing aid fitting. This was achieved by carrying out experiments in which technical (acoustic) differences between alternative hearing aid fittings were absent, while providing test subjects with a strong contrast between the processes apparently applied to derive the fittings being compared. Thus, any effects of contrasting narratives could be observed, free of acoustical confounds. The hypothesis was that narrative effects would be observed. A balanced crossover design was used, in which subjects received and evaluated two bilateral hearing aid fittings in succession. Subjects were deceived as to the true identical content of the hearing aid fittings being compared, but encouraged to believe that one fitting process was "interactive" and the other was "diagnostic" in character. Two almost identical experiments were undertaken: one with 24 experienced adult hearing aid users and another with 16 adult first-time users. Each hearing aid fitting was worn at home for 2 weeks, after which self-report outcome measures (Hearing Aid Performance Questionnaire, Hearing Handicap Inventory for the Elderly, and International Outcome Inventory for Hearing Aids) were administered. After the second test period, a short preference questionnaire was also completed. Twenty of the 24 experienced users showed a clear preference for one or the other fitting, and their self-report scores reflected these preferences. Effect sizes were comparable with those typically observed for true acoustical contrasts. No order effect was seen in this group. In contrast, 13 of the 16 first-time users preferred the second fitting. Trends in the self-report measures were similar for this group but weaker than for the experienced users. In both groups, the reasons given for subjects' preference were predominantly related to sound, despite there being no acoustical differences. This study suggests that the narrative embodied in a given fitting process can have a substantial effect on the perceived benefit of the treatment, independent of any acoustical differences, at least for experienced users. For first-time users, acclimatization seems to overshadow the purely narrative effect of any fitting process. In the future, research study designs should include steps to avoid narrative effects when technical parameters of hearing aids are the intended object of study. In clinical practice, the narrative is part of the therapeutic context, and one may design it for maximum beneficial effect.


Expression of the chloroplast-localized small heat shock protein by oxidative stress in rice.
A rice (Oryza sativa L. cv. Nakdong) cDNA clone, Oshsp26, encoding the chloroplast-localized small heat shock protein (smHSP) was isolated. Southern blot analysis of genomic DNA and the result of screening of a cDNA library indicated that the Oshsp26 gene is encoded by a single gene in the rice genome. The Oshsp26 gene was expressed following heat stress: the transcript level was highest when rice leaves were treated at high temperatures for 2h at 42 degrees C, and the transcripts became detectable after 20min and reached a maximum level after 2h. It was also found that the Oshsp26 gene was expressed following oxidative stress even in the absence of heat stress. Treatment of rice plants with methyl viologen (MV) in the light and treatment with hydrogen peroxide (H(2)O(2)), either in the light or in the dark, both caused a significant accumulation of the transcripts and the protein. Since MV treatment in the light leads to the generation of H(2)O(2) inside the chloroplast, it is likely that H(2)O(2) by itself acts to induce the expression of the Oshsp26 gene. These results suggest that the chloroplast smHSP plays an important role in protecting the chloroplast against damage caused by oxidative stress as well as by heat stress.


Stability of DNA origami nanoarrays in cell lysate.
Scaffolded DNA origami, a method to create self-assembled nanostructures with spatially addressable features, has recently been used to develop water-soluble molecular chips for label-free RNA detection, platforms for deterministic protein positioning, and single molecule reaction observatories. These applications highlight the possibility of exploiting the unique properties and biocompatibility of DNA nanostructures in live, cellular systems. Herein, we assembled several DNA origami nanostructures of differing shape, size and probes, and investigated their interaction with lysate obtained from various normal and cancerous cell lines. We separated and analyzed the origami-lysate mixtures using agarose gel electrophoresis and recovered the DNA structures for functional assay and subsequent microscopic examination. Our results demonstrate that DNA origami nanostructures are stable in cell lysate and can be easily separated from lysate mixtures, in contrast to natural, single- and double-stranded DNA. Atomic force microscope (AFM) and transmission electron microscope (TEM) images show that the DNA origami structures are fully intact after separation from cell lysates and hybridize to their targets, verifying the superior structural integrity and functionality of self-assembled DNA origami nanostructures relative to conventional oligonucleotides. The stability and functionality of DNA origami structures in cell lysate validate their use for biological applications, for example, as programmable molecular rafts or disease detection platforms.


Detection of Mycoplasma genus and Mycoplasma fermentans by PCR in patients with Chronic Fatigue Syndrome.
Mycoplasma fermentans and other Mycoplasma species are colonizers of human mucosal surfaces and may be associated with human immunodeficiency virus infection. While many infectious agents have been described in different percentages of patients with Chronic Fatigue Syndrome (CFS), little is known about the prevalence of mycoplasmas and especially M. fermentans in CFS patients. A polymerase chain reaction (PCR)-based assay was used to detect Mycoplasma genus and M. fermentans genomes in peripheral blood mononuclear cells (PBMC) of CFS patients. Blood was collected from 100 patients with CFS and 50 control subjects. The amplified products of 717 bp of Mycoplasma genus, and 206 bp of M. fermentans were detected in DNA purified from blood samples in 52% and 34% of CFS samples, respectively. In contrast, these genomes were found in only 14% and 8% of healthy control subjects respectively (P < 0.0001). All samples were confirmed by Southern blot with a specific probe based on internal sequences of the expected amplification product. Several samples, which were positive for Mycoplasma genus, were negative for M. fermentans indicating that other Mycoplasma species are involved. A quantitative PCR was developed to determine the number of M. fermentans genome copies present in 1 microg of DNA for controls and CFS patients. Mycoplasma copy numbers ranging from 130 to 880 and from 264 to 2400 were detected in controls and CFS positive subjects, respectively. An enzyme immunoassay was applied for the detection of antibodies against p29 surface lipoprotein of M. fermentans to determine the relationship between M. fermentans genome copy numbers and antibody levels. Individuals with high genome copy numbers exhibited higher IgG and IgM antibodies against M. fermentans specific peptides. Isolation of this organism by culture from clinical specimens is needed in order to demonstrate specificity of signal detected by PCR in this study.


[The surgical treatment of acute ischemia of the lower limb].
To analyze the surgical treatment strategy of lower limb acute ischemia and its results in the Department of Vascular Surgery of Kaunas University of Medicine Hospital in the period from 1999 to 2001. While collecting the data retrospectively there has been determined, that due to the acute ischemia of the limbs there have been operated 244 patients (12.2% of all treated arterial patients). Because of the lower limb acute ischemia there have been operated 142 (7.1%) patients. All embolism and thrombosis have been approved with the operation findings. The comparison of quantitative values was done according to the Student parameter. The acute ischemia in the leg has been caused by: embolism - for 76 (53.5%) patients, acute thrombosis - for 66 (46.5%). Heart diseases have been ascertained in embolism group for 22.4% of patients, in acute thrombosis group - for 6.1%. The localities of embolism - the popliteal artery for 44.7% and femoral artery for 32.9%, acute thrombosis - in the femoral artery for 37.9% and iliac artery for 22.7%. Men experience the acute thrombosis more often (3.5:1), p<0.0001, and women - embolism (2.5:1), p<0.0001. Embolism has been treated with embolectomy, and acute thrombosis with thrombectomy, thrombendarterectomy and the bypass operations. There have been made 4.9% of great amputations, rescued 96.1% of lower limbs with the features of acute ischemia. Total death rate - 4.2%. The acute ischemia in the leg because of embolism was for 76 patients, the acute thrombosis - for 66 patients. Lower limbs embolism of arteries is more common for women, p<0.0001, acute thrombosis for men, p<0.0001. The death rate is higher in women group with acute ischemia than men, (p=0.1198) and in men group with acute ischemia in the lower limb there are made more great amputations (p=0.04250).


Current management of metastatic renal cell carcinoma: evolving new therapies.
Targeted therapies have recently replaced cytokine treatments as the gold standard for management of metastatic renal cell carcinoma (mRCC). Currently approved treatments include the tyrosine kinase inhibitors sunitinib, pazopanib, axitinib, sorafenib, cabozantinib and lenvatinib; the vascular endothelial growth factor (VEGF) inhibitor bevacizumab; the mammalian target of rapamycin (mTOR) inhibitors everolimus and temsirolimus; and the immunologic nivolumab. The purpose of this review is to provide an updated analysis of the clinical data supporting the use of these agents in the first-line and second-line setting. In the first-line setting, pazopanib may be better tolerated than sunitinib, an individualized dosing sunitinib regimen based on toxicity might improve survival and cabozantinib appears to be an emerging option. In the second-line setting, three new therapies (cabozantinib, lenvatinib/everolimus and nivolumab) have shown superiority against everolimus, the previous standard therapy. The International Metastatic RCC Database Consortium prognostic model may be useful in guiding the selection of subsequent therapy and patients eligible for metastasectomy. Targeted therapies are the standard treatment for mRCC. Despite advancements in survival, progression-free survival and tolerability, these targeted therapies remain largely noncurative. Further characterization of the RCC oncogenic pathway, and the ongoing clinical trials should help optimize the management of mRCC.


Primary Ewing sarcoma of the orbit.
To report the clinicopathologic features of a 17-year-old patient with primary Ewing sarcoma of the orbit. The patient was evaluated clinically before surgery with computed tomography scans of the orbit. After surgery, the patient was staged with computed tomography scans and bone scan and was treated with systemic chemotherapy and radiation therapy to the orbit. The orbital biopsy was evaluated with conventional light microscopy and immunohistochemistry. Clinical evaluation revealed proptosis and limited upgaze. Computed tomography scans disclosed a mass involving the superior orbit, anterior cranial fossa, and temporal fossa. Microscopic examination revealed small, poorly differentiated cells with medium-sized nuclei containing finely granular chromatin and small nucleoli. The cytoplasmic borders of the cells were indistinct. A PAS stain revealed modest glycogen in many of the tumor cells. The tumor stained positive for O-13 and vimentin and was negative for neural, skeletal, and lymphoid cell markers. Computed tomography scan, bone scan, and blood chemistries revealed no other site of involvement. After treatment, the clinical symptoms and signs resolved, and there has been no evidence of residual orbital tumor or metastasis. Primary Ewing sarcoma of the orbit should be considered in the differential diagnosis of children or young adults with proptosis, diplopia, or periorbital swelling. Immunohistochemistry is essential to distinguish Ewing sarcoma from other small round cell tumors.


Mutations of the AAAS gene in an Indian family with Allgrove's syndrome.
The triple A or Allgrove's syndrome is an autosomal recessive disorder characterized by the triad of achalasia cardia, alacrima and ACTH resistant adrenocortical insufficiency. Mutations of the Achalasia-Addisonianism-Alacrima-Syndrome (AAAS) gene on chromosome 12q13 are associated with this syndrome. We report an Indian family where two siblings were homozygous for a known mutation of the AAAS gene and presented with the classical triad of symptoms. The mother and the brother were heterozygous and asymptomatic. The affected siblings had iron deficiency anemia and the younger sister had pes cavus and palmoplantar keratosis. Neurological symptoms were absent in both affected children. Recognition of this syndrome can lead to early treatment of adrenal insufficency and genetic counselling.


Congenital cerebral palsy, child sex and parent cardiovascular risk.
Genes associated with cardiovascular disease may also be risk factors for congenital cerebral palsy (CP) and these associations may be modified by sex, since there is an increased risk of CP in male children. We investigated the association between CP of the child with cardiovascular disease in parents, taking sex of the child into consideration. All parents of non-adopted singletons born in Denmark between 1973 and 2003 were included. Parents of a child with CP, confirmed by the Danish National CP registry, were considered exposed. Cox proportional hazards regressions were used to model risk of cardiovascular outcomes for exposed parents compared to all other parents beginning at the child's 10(th) birthday. We identified 733,730 mothers and 666,652 fathers among whom 1,592 and 1,484, respectively, had a child with CP. The mean age for mothers at end of follow up was 50 ± 8 years. After adjustment for maternal age, parental education, child's sex, child's residence, child being small for gestational age and maternal hypertensive disorder during pregnancy, mothers of CP male children had an excess risk of cardiovascular disease (HR: 1.52, 95% CI: 1.16-2.00), attributable mostly to an increased incidence of hypertension and cerebrovascular disease. After additional adjustment for preterm birth, the association was markedly attenuated for cardiovascular disease (1.34, 95%CI: 1.02 - 1.76), became nonsignificant for hypertension, but remained significant for cerebrovascular disease (HR: 2.73, 95% CI: 1.45- 5.12). There was no increased risk of cardiovascular events in mothers of female CP children, or fathers of CP children of any sex. Women that have a male child with CP are at increased risk for premature cardiovascular disease. Part of this association may be related to risk factors for preterm births.


Prejudice Reduction: Progress and Challenges.
The past decade has seen rapid growth in research that evaluates methods for reducing prejudice. This essay reviews 418 experiments reported in 309 manuscripts from 2007 to 2019 to assess which approaches work best and why. Our quantitative assessment uses meta-analysis to estimate average effects. Our qualitative assessment calls attention to landmark studies that are noteworthy for sustained interventions, imaginative measurement, and transparency. However, 76% of all studies evaluate light touch interventions, the long-term impact of which remains unclear. The modal intervention uses mentalizing as a salve for prejudice. Although these studies report optimistic conclusions, we identify troubling indications of publication bias that may exaggerate effects. Furthermore, landmark studies often find limited effects, which suggests the need for further theoretical innovation or synergies with other kinds of psychological or structural interventions. We conclude that much research effort is theoretically and empirically ill-suited to provide actionable, evidence-based recommendations for reducing prejudice.


The HIV1 protein Vpr acts to enhance constitutive DCAF1-dependent UNG2 turnover.
The HIV1 protein Vpr assembles with and acts through an ubiquitin ligase complex that includes DDB1 and cullin 4 (CRL4) to cause G2 cell cycle arrest and to promote degradation of both uracil DNA glycosylase 2 (UNG2) and single-strand selective mono-functional uracil DNA glycosylase 1 (SMUG1). DCAF1, an adaptor protein, is required for Vpr-mediated G2 arrest through the ubiquitin ligase complex. In work described here, we used UNG2 as a model substrate to study how Vpr acts through the ubiquitin ligase complex. We examined whether DCAF1 is essential for Vpr-mediated degradation of UNG2 and SMUG1. We further investigated whether Vpr is required for recruiting substrates to the ubiquitin ligase or acts to enhance its function and whether this parallels Vpr-mediated G2 arrest. We found that DCAF1 plays an important role in Vpr-independent UNG2 and SMUG1 depletion. UNG2 assembled with the ubiquitin ligase complex in the absence of Vpr, but Vpr enhanced this interaction. Further, Vpr-mediated enhancement of UNG2 degradation correlated with low Vpr expression levels. Vpr concentrations exceeding a threshold blocked UNG2 depletion and enhanced its accumulation in the cell nucleus. A similar dose-dependent trend was seen for Vpr-mediated cell cycle arrest. This work identifies UNG2 and SMUG1 as novel targets for CRL4(DCAF1)-mediated degradation. It further shows that Vpr enhances rather than enables the interaction between UNG2 and the ubiquitin ligase. Vpr augments CRL4(DCAF1)-mediated UNG2 degradation at low concentrations but antagonizes it at high concentrations, allowing nuclear accumulation of UNG2. Further, the protein that is targeted to cause G2 arrest behaves much like UNG2. Our findings provide the basis for determining whether the CRL4(DCAF1) complex is alone responsible for cell cycle-dependent UNG2 turnover and will also aid in establishing conditions necessary for the identification of additional targets of Vpr-enhanced degradation.


Leukocyte extravasation: an immunoregulatory role for alpha-L-fucosidase?
Fucosylated oligosaccharides and glycoconjugates have been implicated in several biological events, including the cell-cell adhesion processes that mediate inflammation. Alpha-L-fucosidase (ALF) is an exoglycosidase that is involved in the hydrolytic degradation of alpha-L-fucose from glycoconjugates. In this study, we investigated the potential role of ALF in regulation of leukocyte migration. Measurement of transendothelial migration in response to CCL5 demonstrated that pretreatment of monocytic cells with ALF reduced migration (p = 0.0004) to a greater extent than treatment of the endothelial monolayer (p = 0.0374). Treatment with ALF significantly reduced the adhesion of monocytic cells to immobilized P-selectin.Fc. A murine model of experimental autoimmune uveitis was then used to show that treatment of splenic cells with ALF produced an 8.6-fold decrease in rolling and a 3.2-fold decrease in cell migration across the retinal vasculature. Further in vitro studies demonstrated that treatment of monocytes with the chemokines CCL3 or CCL5 increased the level of mRNA encoding ALF; this was accompanied by the detection of significant increases in both the 51- and 56-kDa components of ALF by Western blotting. Treatment of monocytic cells with ALF for 2 h significantly reduced the cell surface expression of CD31, with a further decrease in expression observed after 5 h (p = 0.002). Thus, CD31 and fucosylated ligands of P-selectin seem to be the candidates through which ALF mediates its effect in vitro. These data identify a previously unrecognized immunoregulatory role for ALF in late stages of inflammation.


Neuromuscular block differentially affects immobility and cortical activation at near-minimum alveolar concentration anesthesia.
Anesthesia-induced immobility and cortical suppression are governed by anatomically separate, but interacting, areas of the central nervous system. Consequently, larger volatile anesthetic concentrations are required to suppress cortical activation than to abolish movement in response to noxious stimulation. We examined the effect of decreased afferent input, as produced by neuromuscular block (NMB), on immobility and cortical activation, as measured by bispectral index (BIS) of the electrocardiogram, in the presence of noxious stimulation during approximately minimum alveolar concentrations (MACs) of desflurane anesthesia. The effect of NMB on the median effective end-tidal concentration of desflurane (EtDes(50), or MAC(tetanus)) for immobility was estimated using the up-and-down method and isolated forearm technique in 24 healthy volunteers. Each volunteer sequentially received saline, mivacurium, and succinylcholine in a randomized order, while EtDes concentration during each of the treatments was determined based on the movement response of the previous volunteer on the same treatment. Nonlinear mixed-effects modeling was used to evaluate the effect of NMB on BIS versus EtDes concentration relationship at baseline and after noxious stimulation, while the frontal electromyogram (EMG(BIS)) effect on BIS was also modeled as a covariate. Cardiovascular responses to noxious stimulation were compared across treatments. Succinylcholine and mivacurium significantly reduced MAC(tetanus) (95% confidence interval) from 5.00% (4.85%-5.13%), during saline, to 4.05% (3.81%-4.29%) and 3.84% (3.60%-4.08%), respectively. Noxious stimulation significantly, although minimally, increased BIS response during all treatments. Succinylcholine increased BIS independently of an effect on EMG(BIS). Succinylcholine administration increased cardiovascular activity. Interestingly, although cardiovascular reaction to the noxious event was ablated by mivacurium, cortical response, as determined by BIS, was retained. Both succinylcholine and mivacurium enhanced immobility during near-MAC anesthesia. All treatments were associated with a small, although significant, BIS increase in response to noxious stimulation, whereas succinylcholine increased BIS independently of noxious stimulation or EMG(BIS). Mivacurium suppressed autonomic response to a noxious event.


Piperonal attenuates visceral adiposity in mice fed a high-fat diet: potential involvement of the adenylate cyclase-protein kinase A dependent pathway.
Piperonal is an aromatic compound found in vanilla and has a floral odor resembling vanillin. This study was aimed to test whether piperonal attenuates visceral adiposity induced by a high-fat diet (HFD) in mice and to explore the underlying molecular mechanisms. Male C57BL/6N mice were fed a normal diet, HFD, or 0.05% piperonal-supplemented HFD (PSD) for 10 weeks. PSD-fed mice showed attenuation of body weight gain, total visceral fat pad weights, and plasma lipid levels compared to HFD-fed mice. Piperonal supplementation of the HFD increased the mRNA expression of certain isotypes of adenylate cyclase (Adcy) and protein kinase A (PKA) in the white adipose tissue (WAT) of mice. The adipogenesis-related genes were downregulated, whereas fatty acid oxidation- and thermogenesis-related genes were upregulated in the WAT of PSD-fed mice compared to those in HFD-fed mice. Piperonal directly activated Adcy by decreasing the Km for its substrate (ATP) in plasma membranes prepared from the WAT of mice. Furthermore, piperonal-induced inhibition of adipocyte differentiation and elevation of Adcy and PKA activities in 3T3-L1 cells were abrogated by an Adcy inhibitor. The anti-adipogenic effect of piperonal in mice fed the high-fat diet appears to be associated with increased Adcy-PKA signaling in WAT.


Overexpression of uridine diphospho glucuronosyltransferase 2B17 in high-risk chronic lymphocytic leukemia.
Uridine diphospho glucuronosyltransferase 2B17 (UGT2B17) glucuronidates androgens and xenobiotics including certain drugs. The UGT2B17 gene shows a remarkable copy number variation (CNV), which predisposes for solid tumors and influences drug response. Here, we identify a yet undescribed UGT2B17 mRNA overexpression in poor-risk chronic lymphocytic leukemia (CLL). In total, 320 CLL patients and 449 healthy donors were analyzed. High (above median) UGT2B17 expression was associated with established CLL poor prognostic factors and resulted in shorter treatment-free and overall survival (hazard ratio ([death] 2.18; 95% CI 1.18-4.01; P = .013). The prognostic impact of mRNA expression was more significant than that of UGT2B17 CNV. UGT2B17 mRNA levels in primary CLL samples directly correlated with functional glucuronidation activity toward androgens and the anticancer drug vorinostat (R > 0.9, P < .001). After treatment with fludarabine containing regimens UGT2B17 was up-regulated particularly in poor responders (P = .030). We observed an exclusive involvement of the 2B17 isoform within the UGT protein family. Gene expression profiling of a stable UGT2B17 knockdown in the CLL cell line MEC-1 demonstrated a significant involvement in key cellular processes. These findings establish a relevant role of UGT2B17 in CLL with functional consequences and potential therapeutic implications.


Contrasting micro/nano architecture on termite wings: two divergent strategies for optimising success of colonisation flights.
Many termite species typically fly during or shortly after rain periods. Local precipitation will ensure water will be present when establishing a new colony after the initial flight. Here we show how different species of termite utilise two distinct and contrasting strategies for optimising the success of the colonisation flight. Nasutitermes sp. and Microcerotermes sp. fly during rain periods and adopt hydrophobic structuring/'technologies' on their wings to contend with a moving canvas of droplets in daylight hours. Schedorhinotermes sp. fly after rain periods (typically at night) and thus do not come into contact with mobile droplets. These termites, in contrast, display hydrophilic structuring on their wings with a small scale roughness which is not dimensionally sufficient to introduce an increase in hydrophobicity. The lack of hydrophobicity allows the termite to be hydrophilicly captured at locations where water may be present in large quantities; sufficient for the initial colonization period. The high wettability of the termite cuticle (Schedorhinotermes sp.) indicates that the membrane has a high surface energy and thus will also have strong attractions with solid particles. To investigate this the termite wings were also interacted with both artificial and natural contaminants in the form of hydrophilic silicon beads of various sizes, 4 µm C(18) beads and three differently structured pollens. These were compared to the superhydrophobic surface of the planthopper (Desudaba psittacus) and a native Si wafer surface. The termite cuticle demonstrated higher adhesive interactions with all particles in comparison to those measured on the plant hopper.


CORL Expression and Function in Insulin Producing Neurons Reversibly Influences Adult Longevity in Drosophila.
CORL proteins (known as SKOR in mice, Fussel in humans and fussel in Flybase) are a family of CNS specific proteins related to Sno/Ski oncogenes. Their developmental and adult roles are largely unknown. A Drosophila CORL (dCORL) reporter gene is expressed in all Drosophila insulin-like peptide 2 (dILP2) neurons of the pars intercerebralis (PI) of the larval and adult brain. The transcription factor Drifter is also expressed in the PI in a subset of dCORL and dILP2 expressing neurons and in several non-dILP2 neurons. dCORL mutant virgin adult brains are missing all dILP2 neurons that do not also express Drifter. This phenotype is also seen when expressing dCORL-RNAi in neurosecretory cells of the PI. dCORL mutant virgin adults of both sexes have a significantly shorter lifespan than their parental strain. This longevity defect is completely reversed by mating (lifespan increases over 50% for males and females). Analyses of dCORL mutant mated adult brains revealed a complete rescue of dILP2 neurons without Drifter. Taken together, the data suggest that dCORL participates in a neural network connecting the insulin signaling pathway, longevity and mating. The conserved sequence and CNS specificity of all CORL proteins imply that this network may be operating in mammals.


Cloning, expression, purification, and crystallisation of HIV-2 reverse transcriptase.
A purification procedure is described for the isolation of recombinant HIV-2 reverse transcriptase expressed in Escherichia coli. The p68 subunit is expressed, in the absence of induction, and use of a heparin-Sepharose column produces substantially pure protein. Concentration of the homodimeric p68 reverse transcriptase pool, followed by incubation at room temperature for several days, results in full conversion by E. coli proteases to the heterodimer (p68/p55). This extended incubation simplifies the purification process and improves the yield of heterodimeric reverse transcriptase, which shows a truncation of the smaller subunit to 427 residues. The protein is then purified further by hydroxyapatite and gel-filtration chromatography to homogeneity. The HIV-2 RT is active and has been used to produce crystals that diffract to beyond 3.0 A.


Versatile and Efficient Site-Specific Protein Functionalization by Tubulin Tyrosine Ligase.
A novel chemoenzymatic approach for simple and fast site-specific protein labeling is reported. Recombinant tubulin tyrosine ligase (TTL) was repurposed to attach various unnatural tyrosine derivatives as small bioorthogonal handles to proteins containing a short tubulin-derived recognition sequence (Tub-tag). This novel strategy enables a broad range of high-yielding and fast chemoselective C-terminal protein modifications on isolated proteins or in cell lysates for applications in biochemistry, cell biology, and beyond, as demonstrated by the site-specific labeling of nanobodies, GFP, and ubiquitin.


Efficacy of a novel online integrated treatment for problem gambling and tobacco smoking: Results of a randomized controlled trial.
Problem gambling and tobacco use are highly comorbid among adults. However, there are few treatment frameworks that target both gambling and tobacco use simultaneously (i.e., an integrated approach), while also being accessible and evidence-based. The aim of this two-arm open label RCT was to examine the efficacy of an integrated online treatment for problem gambling and tobacco use. A sample of 209 participants (Mage = 37.66, SD = 13.81; 62.2% female) from North America were randomized into one of two treatment conditions (integrated [n = 91] or gambling only [n = 118]) that lasted for eight weeks and consisted of seven online modules. Participants completed assessments at baseline, after treatment completion, and at 24-week follow-up. While a priori planned generalized linear mixed models showed no condition differences on primary (gambling days, money spent, time spent) and secondary outcomes, both conditions did appear to significantly reduce problem gambling and smoking behaviours over time. Post hoc analyses showed that reductions in smoking and gambling craving were correlated with reductions in days spent gambling, as well as with gambling disorder symptoms. Relatively high (versus low) nicotine replacement therapy use was associated with greater reductions in gambling behaviours in the integrated treatment condition. While our open label RCT does not support a clear benefit of integrated treatment, findings suggest that changes in smoking and gambling were correlated over time, regardless of treatment condition, suggesting that more research on mechanisms of smoking outcomes in the context of gambling treatment may be relevant.


Placental pathology in pregnancy-induced hypertension (PIH) with or without intrauterine growth retardation.
The foetus, placenta and mother form a composite triad of dynamic equilibrium, and dysfunction of any one of them can affect the others. The aim of the present study was to appreciate qualitatively and to assess quantitatively the pathological features of placentas associated with pregnancy induced hypertension (PIH) with and without intrauterine growth retardation (IUGR) & also IUGR placentas without PIH and to compare them with the normal placentas and to evolve plausible explanation for the outcomes of PIH and IUGR. The study evaluated 40, 51, 58 placentas of PIH with IUGR (category I), PIH without IUGR (category II), IUGR without PIH (category III) respectively and compared them with 118 normal placentas. The basic conclusions of the study were that PIH is common among primigravida and probably the main factor in the genesis of IUGR and reduced placental weight. There was a high prevalence of inflammation, infarction, ischemia, intervillous hemorrhage, and syncytial knots in PIH with IUGR placentas as also in PIH without IUGR (category II) and IUGR without PIH (category III) placentas. Acute atherosis was a characteristic finding of PIH placentas (both category I & II).


Epidemiology/risk factors of sexual dysfunction.
Accurate estimates of prevalence/incidence are important in understanding the true burden of male and female sexual dysfunction and in identifying risk factors for prevention efforts. To provide recommendations/guidelines concerning state-of-the-art knowledge for the epidemiology/risk factors of sexual dysfunctions in men and women. An International Consultation in collaboration with the major urology and sexual medicine associations assembled over 200 multidisciplinary experts from 60 countries into 17 committees. Committee members established specific objectives and scopes for various male and female sexual medicine topics. The recommendations concerning state-of-the-art knowledge in the respective sexual medicine topic represent the opinion of experts from five continents developed in a process over a 2-year period. Concerning the Epidemiology/Risk Factors Committee, there were seven experts from four countries. Expert opinion was based on grading of evidence-based medical literature, widespread internal committee discussion, public presentation and debate. Standard definitions of male and female sexual dysfunctions are needed. The incidence rate for erectile dysfunction is 25-30 cases per thousand person years and increases with age. There are no parallel data for women's sexual dysfunctions. The prevalence of sexual dysfunction increases as men and women age; about 40-45% of adult women and 20-30% of adult men have at least one manifest sexual dysfunction. Common risk factor categories associated with sexual dysfunction exist for men and women including: individual general health status, diabetes mellitus, cardiovascular disease, other genitourinary disease, psychiatric/psychological disorders, other chronic diseases, and socio-demographic conditions. Endothelial dysfunction is a condition present in many cases of erectile dysfunction and there are common etiological pathways for other vascular disease states. Increasing physical activity lowers incidence of ED in males who initiate follow-up in their middle ages. There is a need for more epidemiologic research in male and female sexual dysfunction.


Intranodal vaccination with naked antigen-encoding RNA elicits potent prophylactic and therapeutic antitumoral immunity.
Although naked antigen-encoding RNA has entered clinical testing, basic knowledge on how to apply this promising novel vaccine format is still pending. By comparing different administration routes, we observed surprisingly potent antigen-specific T-cell immunity upon intranodal injection of naked antigen-encoding RNA. RNA was selectively uptaken by resident dendritic cells, propagated a T-cell attracting and stimulatory intralymphatic milieu, and led to efficient expansion of antigen-specific CD8+ as well as CD4+ T cells. By intranodal treatment of mice with repeated cycles of RNA, we achieved de novo priming of naïve T cells, which became potent cytolytic effectors capable of homing to primary and secondary lymphatic tissues as well as memory T cells. In tumor-bearing mice intralymphatic RNA vaccination elicited protective and therapeutic antitumor immune responses, resulting in a remarkable survival benefit as compared with other treatment regimens. This is the first report of strong systemic antigen-specific Th1-type immunity and cancer cure achieved with naked antigen-encoding RNA in preclinical animal models.


Venous thromboembolic disease: an observational study in medical-surgical intensive care unit patients.
Acute and chronic illness, immobility, and procedural and pharmacologic interventions may predispose patients in the intensive care unit (ICU) to venous thromboembolic (VTE) disease. The purpose of this study was to observe potential risk factors and diagnostic tests for VTE, and prophylaxis against VTE in medical-surgical ICU patients. In a prospective observational study, 93 consecutive patients admitted to a mixed medical-surgical ICU were followed. We recorded demographics, admitting diagnoses, APACHE II score, VTE risk factors, antithrombotic, anticoagulant and thrombolytic agents, diagnostic tests for deep venous thrombosis (DVT) and pulmonary embolus (PE), and clinical outcomes. Patients were 65.5 (15.5) years old with an APACHE II score of 21.1 (9.0); 44 (47.3%) were female. Admission diagnoses were medical (58, 67.4%) and surgical (35, 37.6%). The duration of ICU stay was 3 days (interquartile range: 1, 8.5 days) and the ICU mortality rate was 20.4% (19 of 93). We observed 8 VTE events among 5 of 93 patients (incidence 5.4% [0.8 to 10.0]); 2 patients had DVT and PE before admission, 1 had DVT as an admitting diagnosis, 1 had DVT on day 2 and PE on day 3, and 1 had PE on day 2. Over 804 ICU patient-days, 2 of 5 ultrasound examinations diagnosed DVT and 2 of 3 ventilation-perfusion lung scans diagnosed PE. Of 64 patients in whom heparin was not contraindicated and who were not anticoagulated, subcutaneous heparin prophylaxis was prescribed for 40 (62.5%) patients. ICU-acquired VTE risk factors were mechanical ventilation (odds ratio [OR] 1.56), immobility (OR 2.14), femoral venous catheter (OR 2.24), sedatives (OR 1.52), and paralytic drugs (OR 4.81), whereas VTE heparin prophylaxis (OR 0.08), aspirin (OR 0.42), and thromboembolic disease stockings (OR 0.63) were associated with a lower risk. Only warfarin (OR 0.07, P =.01) and intravenous heparin (OR 0.04, P<.01) were associated with a significantly decreased risk of VTE. Several ICU-acquired risk factors for VTE were documented in this medical-surgical ICU. VTE prophylaxis was underprescribed, and VTE diagnostic tests were infrequent. Further research is required to determine the incidence, predisposing factors, attributable morbidity, mortality, and costs of VTE in medical-surgical ICU patients, the optimal diagnostic test strategies, and the most cost-effective approaches of prophylaxis.


Evaluating Darwin's naturalization hypothesis in experimental plant assemblages: phylogenetic relationships do not determine colonization success.
Darwin's naturalization hypothesis (DNH) proposes that colonization is less likely when the colonizing species is related to members of the invaded community, because evolutionary closeness intensifies competition among species that share similar resources. Studies that have evaluated DNH from correlational evidence have yielded controversial results with respect to its occurrence and generality. In the present study we carried out a set of manipulative experiments in which we controlled the phylogenetic relatedness of one colonizing species (Lactuca sativa) with five assemblages of plants (the recipient communities), and evaluated the colonizing success using five indicators (germination, growth, flowering, survival, and recruitment). The evolutionary relatedness was calculated as the mean phylogenetic distance between Lactuca and the members of each assemblage (MPD) and by the mean phylogenetic distance to the nearest neighbor (MNND). The results showed that the colonization success of Lactuca was not affected by MPD or MNND values, findings that do not support DNH. These results disagree with experimental studies made with communities of microorganisms, which show an inverse relation between colonization success and phylogenetic distances. We suggest that these discrepancies may be due to the high phylogenetic distance used, since in our experiments the colonizing species (Lactuca) was a distant relative of the assemblage members, while in the other studies the colonizing taxa have been related at the congeneric and conspecific levels. We suggest that under field conditions the phylogenetic distance is a weak predictor of competition, and it has a limited role in determining colonization success, contrary to prediction of the DNH. More experimental studies are needed to establish the importance of phylogenetic distance between colonizing species and invaded community on colonization success.


Cardiorespiratory responses during cycle ergometer exercise with different ramp slope increments in patients with chronic obstructive pulmonary disease.
The ramp exercise test has been widely used to evaluate cardiopulmonary responses to an incremental exercise load. This study was performed to clarify whether different slopes of the ramp exercise test influence exercise tolerance, exercise limiting factors, and respiratory pattern in patients with chronic obstructive pulmonary disease (COPD). We applied three different slopes (5 W/min, 10 W/min and 20 W/min) of the ramp exercise test in 9 patients with COPD and evaluated cardiopulmonary responses. There were no significant differences in peak oxygen uptake, anaerobic threshold (AT), minute ventilation, heart rate, arterial oxygen saturation, expired tidal volume, or respiratory rate at the maximal load among the three different ramp exercises tested. AT could be determined in six of nine patients (67%) at the slope of 5 W/min, in 8/9 (89%) at the slope of 10 W/min, and in 9/9 (100%) at the slope of 20 W/min. The findings suggest that the ramp slope does not affect exercise tolerance, exercise limiting factors, or respiratory patterns and each of these ramp slopes is useful for the evaluation of COPD. Ramp slopes of 10 W/ min or 20 W/min should be appropriate for the determination of AT.


Towards the proteome of Burkholderia cenocepacia H111: setting up a 2-DE reference map.
Polyphasic-taxonomic studies of the past decade have shown that the Burkholderia cepacia complex (Bcc) comprises at least nine species, which share a high degree of 16S rDNA (98-100%) sequence similarity but only moderate levels of DNA-DNA hybridization. Members of the Bcc are well known as opportunistic pathogens of plants, animals and humans but also as biocontrol and bioremediation agents. In this study intra-, surface-associated and extracellular proteins of B. cenocepacia H111, which was isolated from a cystic fibrosis patient, were examined by 2-DE coupled to MALDI-TOF MS. MS and MS/MS data were searched against a database comprising all currently available annotated proteins of genetically closely related strains. In total 642 proteins spots were successfully identified corresponding to 390 different protein species, which were classified into functional categories. The majority of these proteins could be linked to housekeeping functions in energy production, amino acid metabolism, protein folding, post-translational modification and turnover, and translation. Noteworthy is the fact that a significant number of truly secreted and membrane proteins were identified in the extracellular and surface-associated sub-proteomes. This indicates that the pre-fractionation protocol used in this study is a highly valuable strategy for unravelling the cellular location of the identified proteins.


A 12-month phase 3 study of pasireotide in Cushing's disease.
Cushing's disease is associated with high morbidity and mortality. Pasireotide, a potential therapy, has a unique, broad somatostatin-receptor-binding profile, with high binding affinity for somatostatin-receptor subtype 5. In this double-blind, phase 3 study, we randomly assigned 162 adults with Cushing's disease and a urinary free cortisol level of at least 1.5 times the upper limit of the normal range to receive subcutaneous pasireotide at a dose of 600 μg (82 patients) or 900 μg (80 patients) twice daily. Patients with urinary free cortisol not exceeding 2 times the upper limit of the normal range and not exceeding the baseline level at month 3 continued to receive their randomly assigned dose; all others received an additional 300 μg twice daily. The primary end point was a urinary free cortisol level at or below the upper limit of the normal range at month 6 without an increased dose. Open-label treatment continued through month 12. Twelve of the 82 patients in the 600-μg group and 21 of the 80 patients in the 900-μg group met the primary end point. The median urinary free cortisol level decreased by approximately 50% by month 2 and remained stable in both groups. A normal urinary free cortisol level was achieved more frequently in patients with baseline levels not exceeding 5 times the upper limit of the normal range than in patients with higher baseline levels. Serum and salivary cortisol and plasma corticotropin levels decreased, and clinical signs and symptoms of Cushing's disease diminished. Pasireotide was associated with hyperglycemia-related adverse events in 118 of 162 patients; other adverse events were similar to those associated with other somatostatin analogues. Despite declines in cortisol levels, blood glucose and glycated hemoglobin levels increased soon after treatment initiation and then stabilized; treatment with a glucose-lowering medication was initiated in 74 of 162 patients. The significant decrease in cortisol levels in patients with Cushing's disease who received pasireotide supports its potential use as a targeted treatment for corticotropin-secreting pituitary adenomas. (Funded by Novartis Pharma; ClinicalTrials.gov number, NCT00434148.).


Understanding the adaptation of Halobacterium species NRC-1 to its extreme environment through computational analysis of its genome sequence.
The genome of the halophilic archaeon Halobacterium sp. NRC-1 and predicted proteome have been analyzed by computational methods and reveal characteristics relevant to life in an extreme environment distinguished by hypersalinity and high solar radiation: (1) The proteome is highly acidic, with a median pI of 4.9 and mostly lacking basic proteins. This characteristic correlates with high surface negative charge, determined through homology modeling, as the major adaptive mechanism of halophilic proteins to function in nearly saturating salinity. (2) Codon usage displays the expected GC bias in the wobble position and is consistent with a highly acidic proteome. (3) Distinct genomic domains of NRC-1 with bacterial character are apparent by whole proteome BLAST analysis, including two gene clusters coding for a bacterial-type aerobic respiratory chain. This result indicates that the capacity of halophiles for aerobic respiration may have been acquired through lateral gene transfer. (4) Two regions of the large chromosome were found with relatively lower GC composition and overrepresentation of IS elements, similar to the minichromosomes. These IS-element-rich regions of the genome may serve to exchange DNA between the three replicons and promote genome evolution. (5) GC-skew analysis showed evidence for the existence of two replication origins in the large chromosome. This finding and the occurrence of multiple chromosomes indicate a dynamic genome organization with eukaryotic character.


Axillary tumour burden in women with one abnormal node on ultrasound compared to women with multiple abnormal nodes.
To determine if the number of abnormal nodes seen on preoperative axillary ultrasound (AUS) is a predictor of the number of positive nodes at histology for women with needle-biopsy-proven positive nodes. This prospective multicentre cohort study included consecutive patients with early breast cancer who had needle-biopsy-proven positive nodes on AUS and underwent axillary lymph node dissection (ALND) between October 2015 and July 2016. The number of abnormal nodes at preoperative AUS was recorded by breast radiologists or radiographers. One hundred and twenty-three patients were included in the study. The median age of the women was 62 (range 30-93) years. Fifty-four of the 123 (44%) women had one abnormal node, whereas 69 (56%) had multiple abnormal nodes on AUS. Forty of the 123 (33%) women had two or fewer nodes with metastases at histology after ALND. Tumours ≤20 mm (p<0.001) and one abnormal node on AUS (p<0.001) were associated with two or fewer nodes with metastases at ALND. Both remained significant in logistic regression analysis. The likelihood of at least three metastases based on the combination of these two factors had 95% sensitivity (79 of 83), 35% specificity (14 of 40), a negative predictive value of 78% (14 of 18), and a positive predictive value of 75% (79 of 105). Among women with needle-biopsy-proven positive nodes, around three in four women (78%) with an invasive tumour ≤2 cm and one abnormal node on AUS have two or fewer positive nodes at ALND. These women are overtreated by upfront ALND and can be offered sentinel node biopsy (SNB).


Chronic myeloid leukemia in adolescents and young adults: patient characteristics, outcomes and review of the literature.
Over the past two decades, many improvements have been made in the management of patients with leukemia. Research in this field most often focuses on the youngest and oldest patient age groups. However, the population of patients in between those age groups has received relatively little attention with few studies specifically focusing on them. This important 'age gap' has demonstrated a unique, difficult-to-treat group of patients known as adolescents and young adults, or AYAs. Variably defined in the literature as patients from late teenage years to the age of up to 40 years, the AYA group of patients represents a vulnerable subset of patients now identified to require its own focus, development of therapeutic strategies and parallel emphasis on special support systems involving multidisciplinary psychosocial care. Despite the great advancements that have been realized for patients with chronic myeloid leukemia (CML), the AYA group has seldom been the focus of specific reports and studies, and the outcome appears to lag behind the general population. This review focuses on this subset of AYA patients with CML and summarizes the available data and recent developments, challenges and treatment options for this group of patients.


Loneliness is closely associated with depression outcomes and suicidal ideation among military veterans in primary care.
Although the substantial influence of social relationships on health is well-known, studies that concurrently examine the influence of varying dimensions of social connectedness on major depression are more limited. This study's aim was to determine to what degree several facets of social connectedness (number of confidants, social support, interpersonal conflict, social norms, and loneliness) are correlated with depression-related outcomes. Participants were primary care patients (n = 301) with probable major depression at a Veterans Health Administration hospital and its satellite clinics. Social connectedness was primarily measured using multi-item instruments from the NIH Toolbox of Adult Social Relationship Scales. Primary outcomes were clinical symptoms (depression and suicidal ideation) and secondary outcomes were self-reported health-related behaviors (medication adherence, patient activation, and help-seeking intentions). In multivariate models adjusting for potential confounders and other facets of connectedness, loneliness was associated with higher levels of depression and suicidal ideation, as well as lower patient activation and help-seeking intentions. Social support and social norms about depression treatment were each associated with higher patient activation and help-seeking intentions. Social connectedness was not associated with medication adherence. The limitations of this study are primarily related to its cross-sectional survey design and study population. Multiple aspects of social connectedness are associated with depression outcomes among military veterans with depression. Loneliness may represent the most important component of connectedness, as it is associated with depression severity, suicidality, and health-related behaviors.


The Effects of Active IBD During Pregnancy in the Era of Novel IBD Therapies.
Previous data on inflammatory bowel disease (IBD) relapse during pregnancy mainly originate from retrospective studies. The aim of this study was therefore (i) to evaluate the effect of active disease at conception and IBD disease type on disease relapse during pregnancy and (ii) to study the effects of disease relapse during pregnancy on birth outcomes in a prospective cohort with adequate representation of current treatments. From 2008 to 2014, IBD women were recruited from an ongoing prospective clinical cohort. All patients with confirmed IBD diagnosis with a pregnancy wish or pregnancy were prospectively followed-up until pregnancy and delivery. Disease relapse was measured at each visit. Birth outcomes were recorded from the obstetrician. A total of 298 pregnancies were observed in 229 IBD patients (157 Crohn's disease (CD), 66 ulcerative colitis (UC), and 6 IBD unclassified), resulting in 226 live births. Active disease at conception was strongly associated with disease relapse during pregnancy (aOR=7.66, 95% confidence interval (CI): 3.77-15.54). UC patients experienced relapse during pregnancy more often than CD patients, independent of maternal age, smoking, periconceptional disease activity, previous IBD surgery, and the use of immunosuppressives or anti-tumor necrosis factor (TNF) (aOR=3.71, 95% CI:1.86-7.40). Disease relapse was not associated with adverse birth outcomes such as spontaneous abortion, low-birth weight, or preterm birth. This study confirms that active disease around conception increases the risk of disease relapse during pregnancy. In addition, UC patients relapse more often during pregnancy than CD patients. Birth outcomes were excellent, reflecting the stringent follow-up and treatment of this group of patients.


Dopamine response gene pathways in dorsal striatum MSNs from a gene expression viewpoint: cAMP-mediated gene networks.
Medium spiny neurons (MSNs) comprise the main body (95% in mouse) of the dorsal striatum neurons and represent dopaminoceptive GABAergic neurons. The cAMP (cyclic Adenosine MonoPhosphate)-mediated cascade of excitation and inhibition responses observed in MSN intracellular signal transduction is crucial for neuroscience research due to its involvement in the motor and behavioral functions. In particular, all types of addictions are related to MSNs. Shedding the light on the mechanics of the above-mentioned cascade is of primary importance for this research domain. A mouse model of chronic social conflicts in daily agonistic interactions was used to analyze dorsal striatum neurons genes implicated in cAMP-mediated phosphorylation activation pathways specific for MSNs. Based on expression correlation analysis, we succeeded in dissecting Drd1- and Drd2-dopaminoceptive neurons (D1 and D2, correspondingly) gene pathways. We also found that D1 neurons genes clustering are split into two oppositely correlated states, passive and active ones, the latter apparently corresponding to D1 firing stage upon protein kinase A (PKA) activation. We observed that under defeat stress in chronic social conflicts the loser mice manifest overall depression of dopamine-mediated MSNs activity resulting in previously reported reduced motor activity, while the aggressive mice with positive fighting experience (aggressive mice) feature an increase in both D1-active phase and D2 MSNs genes expression leading to hyperactive behavior pattern corresponded by us before. Based on the alternative transcript isoforms expression analysis, it was assumed that many genes (Drd1, Adora1, Pde10, Ppp1r1b, Gnal), specifically those in D1 neurons, apparently remain transcriptionally repressed via the reversible mechanism of promoter CpG island silencing, resulting in alternative promoter usage following profound reduction in their expression rate. Based on the animal stress model dorsal striatum pooled tissue RNA-Seq data restricted to cAMP related genes subset we elucidated MSNs steady states exhaustive projection for the first time. We correspond the existence of D1 active state not explicitly outlined before, and connected with dynamic dopamine neurotransmission cycles. Consequently, we were also able to indicate an oscillated postsynaptic dopamine vs glutamate action pattern in the course of the neurotransmission cycles.


Vernier learning with short- and long-staircase training and its transfer to a new location with double training.
We previously demonstrated that perceptual learning of Vernier discrimination, when paired with orientation learning at the same retinal location, can transfer completely to untrained locations (Wang, Zhang, Klein, Levi, & Yu, 2014; Zhang, Wang, Klein, Levi, & Yu, 2011). However, Hung and Seitz (2014) reported that the transfer is possible only when Vernier is trained with short staircases, but not with very long staircases. Here we ran two experiments to examine Hung and Seitz's conclusions. The first experiment confirmed the transfer effects with short-staircase Vernier training in both our study and Hung and Seitz's. The second experiment revealed that long-staircase training only produced very fast learning at the beginning of the pretraining session, but with no further learning afterward. Moreover, the learning and transfer effects differed insignificantly with a small effect size, making it difficult to support Hung and Seitz's claim that learning with long-staircase training cannot transfer to an untrained retinal location.


Microengineered cultures containing human hepatic stellate cells and hepatocytes for drug development.
In non-alcoholic steatohepatitis (NASH), hepatic stellate cells (HSC) differentiate into myofibroblast-like cells that cause fibrosis, which predisposes patients to cirrhosis and hepatocellular carcinoma. Thus, modeling interactions between activated HSCs and hepatocytes in vitro can aid in the development of anti-NASH/fibrosis therapeutics and lead to a better understanding of disease progression. Species-specific differences in drug metabolism and disease pathways now necessitate the supplementation of animal studies with data acquired using human liver models; however, current models do not adequately model the negative effects of primary human activated HSCs on the phenotype of otherwise well-differentiated primary human hepatocytes (PHHs) as in vivo. Therefore, here we first determined the long-term effects of primary human activated HSCs on PHH phenotype in a micropatterned co-culture (MPCC) platform while using 3T3-J2 murine embryonic fibroblasts as the control cell type since it has been shown previously to stabilize PHH functions for 4-6 weeks. We found that HSCs were not able to stabilize the PHH phenotype to the same magnitude and longevity as the fibroblasts, which subsequently inspired the development of a micropatterned tri-culture (MPTC) platform in which (a) micropatterned PHHs were functionally stabilized using fibroblasts, and (b) the PHH phenotype was modulated by culturing HSCs within the fibroblast monolayer at physiologically-relevant ratios with PHHs. Transwell inserts containing HSCs were placed atop MPCCs containing fibroblasts to confirm the effects of paracrine signaling between PHHs and HSCs. We found that while albumin and urea secretions were relatively similar in MPTCs and MPCCs (suggesting well-differentiated PHHs), increasing HSC numbers within MPTCs downregulated hepatic cytochrome-P450 (2A6, 3A4) and transporter activities, and caused steatosis over 2 weeks. Furthermore, MPTCs secreted higher levels of pro-inflammatory interleukin-6 (IL-6) cytokine and C-reactive protein (CRP) than MPCCs. Treatment of MPCCs with HSC-conditioned culture medium confirmed that HSC secretions mediate the altered phenotype of PHHs observed in MPTCs, partly via IL-6 signaling. Lastly, we found that NADPH oxidase (NOX) inhibition and farnesoid X receptor (FXR) activation using clinically relevant drugs alleviated hepatic dysfunctions in MPTCs. In conclusion, MPTCs recapitulate symptoms of NASH- and early fibrosis-like dysfunctions in PHHs and have utility for drug discovery in this space.


FBXO11 targets BCL6 for degradation and is inactivated in diffuse large B-cell lymphomas.
BCL6 is the product of a proto-oncogene implicated in the pathogenesis of human B-cell lymphomas. By binding specific DNA sequences, BCL6 controls the transcription of a variety of genes involved in B-cell development, differentiation and activation. BCL6 is overexpressed in the majority of patients with aggressive diffuse large B-cell lymphoma (DLBCL), the most common lymphoma in adulthood, and transgenic mice constitutively expressing BCL6 in B cells develop DLBCLs similar to the human disease. In many DLBCL patients, BCL6 overexpression is achieved through translocation (~40%) or hypermutation of its promoter (~15%). However, many other DLBCLs overexpress BCL6 through an unknown mechanism. Here we show that BCL6 is targeted for ubiquitylation and proteasomal degradation by a SKP1–CUL1–F-box protein (SCF) ubiquitin ligase complex that contains the orphan F-box protein FBXO11 (refs 5, 6). The gene encoding FBXO11 was found to be deleted or mutated in multiple DLBCL cell lines, and this inactivation of FBXO11 correlated with increased levels and stability of BCL6. Similarly, FBXO11 was either deleted or mutated in primary DLBCLs. Notably, tumour-derived FBXO11 mutants displayed an impaired ability to induce BCL6 degradation. Reconstitution of FBXO11 expression in FBXO11-deleted DLBCL cells promoted BCL6 ubiquitylation and degradation, inhibited cell proliferation, and induced cell death. FBXO11-deleted DLBCL cells generated tumours in immunodeficient mice, and the tumorigenicity was suppressed by FBXO11 reconstitution. We reveal a molecular mechanism controlling BCL6 stability and propose that mutations and deletions in FBXO11 contribute to lymphomagenesis through BCL6 stabilization. The deletions/mutations found in DLBCLs are largely monoallelic, indicating that FBXO11 is a haplo-insufficient tumour suppressor gene.


Characterization of human pseudogene-derived non-coding RNAs for functional potential.
Thousands of pseudogenes exist in the human genome and many are transcribed, but their functional potential remains elusive and understudied. To explore these issues systematically, we first developed a computational pipeline to identify transcribed pseudogenes from RNA-Seq data. Applying the pipeline to datasets from 16 distinct normal human tissues identified ∼ 3,000 pseudogenes that could produce non-coding RNAs in a manner of low abundance but high tissue specificity under normal physiological conditions. Cross-tissue comparison revealed that the transcriptional profiles of pseudogenes and their parent genes showed mostly positive correlations, suggesting that pseudogene transcription could have a positive effect on the expression of their parent genes, perhaps by functioning as competing endogenous RNAs (ceRNAs), as previously suggested and demonstrated with the PTEN pseudogene, PTENP1. Our analysis of the ENCODE project data also found many transcriptionally active pseudogenes in the GM12878 and K562 cell lines; moreover, it showed that many human pseudogenes produced small RNAs (sRNAs) and some pseudogene-derived sRNAs, especially those from antisense strands, exhibited evidence of interfering with gene expression. Further integrated analysis of transcriptomics and epigenomics data, however, demonstrated that trimethylation of histone 3 at lysine 9 (H3K9me3), a posttranslational modification typically associated with gene repression and heterochromatin, was enriched at many transcribed pseudogenes in a transcription-level dependent manner in the two cell lines. The H3K9me3 enrichment was more prominent in pseudogenes that produced sRNAs at pseudogene loci and their adjacent regions, an observation further supported by the co-enrichment of SETDB1 (a H3K9 methyltransferase), suggesting that pseudogene sRNAs may have a role in regional chromatin repression. Taken together, our comprehensive and systematic characterization of pseudogene transcription uncovers a complex picture of how pseudogene ncRNAs could influence gene and pseudogene expression, at both epigenetic and post-transcriptional levels.


Bistable bacterial growth rate in response to antibiotics with low membrane permeability.
We demonstrate that growth rate bistability for bacterial cells growing exponentially at a fixed external antibiotic concentration can emerge when the cell wall permeability for the drug is low and the growth rate sensitivity to the intracellular drug concentration is high. Under such conditions, an initially high growth rate can remain high, due to dilution of the intracellular drug concentration by rapid cell volume increase, while an initially low growth rate can remain low, due to slow cell volume increase and insignificant drug dilution. Our findings have implications for the testing of novel antibiotics on growing bacterial strains.


Novel breast cancer risk alleles and interaction with ionizing radiation among U.S. radiologic technologists.
As genome-wide association studies of breast cancer are replicating findings and refinement studies are narrowing the signal location, additional efforts are necessary to elucidate the underlying functional relationships. One approach is to evaluate variation in risk by genotype based on known breast carcinogens, such as ionizing radiation. Given the public health concerns associated with recent increases in medical radiation exposure, this approach may also identify potentially susceptible subpopulations. We examined interaction between 27 newly identified breast cancer risk alleles (identified within the NCI Cancer Genetic Markers of Susceptibility and the Breast Cancer Association Consortium genome-wide association studies) and occupational and medical diagnostic radiation exposure among 859 cases and 1083 controls nested within the United States Radiologic Technologists cohort. We did not find significant variation in the radiation-related breast cancer risk for the variant in RAD51L1 (rs10483813) on 14q24.1 as we had hypothesized. In exploratory analyses, we found that the radiation-associated breast cancer risk varied significantly by linked markers in 5p12 (rs930395, rs10941679, rs2067980 and rs4415084) in the mitochondrial ribosomal protein S30 (MRPS30) gene (P(interaction) = 0.04). Chance, however, may explain these findings, and as such, these results need to be confirmed in other populations with low to moderate levels of radiation exposure. Even though a complete understanding of the way(s) in which these variants may increase breast cancer risk remains elusive, this approach may yield clues for further investigation.


The Home Literacy Environment Is a Correlate, but Perhaps Not a Cause, of Variations in Children's Language and Literacy Development.
The home literacy environment is a well-established predictor of children's language and literacy development. We investigated whether formal, informal, and indirect measures of the home literacy environment predict children's reading and language skills once maternal language abilities are taken into account. Data come from a longitudinal study of children at high risk of dyslexia (N = 251) followed from preschool years. Latent factors describing maternal language were significant predictors of storybook exposure but not of direct literacy instruction. Maternal language and phonological skills respectively predicted children's language and reading/spelling skills. However, after accounting for variations in maternal language, storybook exposure was not a significant predictor of children's outcomes. In contrast, direct literacy instruction remained a predictor of children's reading/spelling skills. We argue that the relationship between early informal home literacy activities and children's language and reading skills is largely accounted for by maternal skills and may reflect genetic influences.


Timely Family Feedback to Guide Family Engagement in the Intensive Care Unit.
Organizations motivated to provide high-quality care in the intensive care unit are exploring strategies to engage families in patient care. Such initiatives are based on emerging evidence that family engagement improves quality and safety of care. To gather family feedback to guide future nurse-led quality improvement efforts to engage families in the intensive care unit setting. The Critical Care Family Satisfaction Survey, which consists of 20 items rated from 1 (very dissatisfied) to 5 (very satisfied), was paired with open-ended questions and administered to families during the intensive care unit stay from March through December 2017. Content analysis was used to identify themes regarding the family experience. Responses were collected from 178 family members. The mean (SD) score on the survey was 4.65 (0.33). Five themes emerged regarding the delivery of family care in the intensive care unit: family interactions with the interdisciplinary team, information sharing and effective communication, family navigation of the intensive care unit environment, family engagement in the intensive care unit, and quality of patient care. This quality improvement project provided foundational information to guide family engagement efforts in the intensive care unit. Real-time solicitation of feedback is essential to improving the family experience and guiding family-centered care delivery in this practice environment.


The rate of bactericidal action of penicillin in vitro as a function of its concentration, and its paradoxically reduced activity at high concentrations against certain organisms.
1. The concentrations of penicillin G which (a) reduced the net rate of multiplication, (b) exerted a net bactericidal effect, and (c) killed the organisms at a maximal rate, have been defined for a total of 41 strains of alpha- and beta-hemolytic streptococci, Staphylococcus aureus and Staphylococcus albus, Diplococcus pneumoniae, and the Reiter treponoma. 2. The concentration which killed the organisms at a maximal rate was 2 to 20 times the minimal effective level ("sensitivity" as ordinarily defined). With some organisms, even a 32,000-fold increase beyond this maximally effective level did not further increase the rate of its bactericidal effect. However, with approximately half the strains here studied (all 4 strains of group B beta-hemolytic streptococci, 4 of 5 group C strains, 5 of 7 strains of Streptococcus fecalis, 2 of 4 other alpha-hemolytic streptococci, and 4 of 9 strains of staphylococci), when the concentration of penicillin was increased beyond that optimal level, the rate at which the organisms died was paradoxically reduced rather than increased, so that the maximal effect was obtained only within a relatively narrow optimal zone. 3. There were marked differences between bacterial species, and occasionally between different strains of the same species, not only with respect to the effective concentrations of penicillin, but also with respect to the maximal rate at which they could be killed by the drug in any concentration. Although there was a rough correlation between these two factors, there were many exceptions; individual strains affected only by high concentrations of penicillin might nevertheless be killed rapidly, while strains sensitive to minute concentrations might be killed only slowly. 4. Within the same bacterial suspension, individual organisms varied only to a minor degree with respect to the effective concentrations of penicillin. They varied strikingly, however, in their resistance to penicillin as measured by the times required to kill varying proportions of the cells.


Antibiofilm activity of α-mangostin extracted from Garcinia mangostana L. against Staphylococcus aureus.
To isolate α-mangostin (AMG) from the peels of mangosteen (Garcinia mangostana L.), grown in Vietnam, and to investigate antibiofilm activity of this compound against three Staphylococcus aureus (S. aureus) strains, one of which was methicillin-resistant S. aureus (MRSA) and the other two strains were methicillin-sensitive S. aureus (MSSA). AMG in n-hexane fraction was isolated on a silica gel column and chemically analyzed by HPLC and NMR. The antibiofilm activity of this compound was investigated by using a 96-well plate model for the formation of biofilms. Biofilm biomass was quantified using crystal violet. The viability of cells was observed under confocal microscopy using LIVE/DEAD BacLight stains. Biofilm composition was determined using specific chemical and enzyme tests for polysaccharide, protein and DNA. Membrane-damaging activity was assayed by measuring the hemolysis of human red blood cells in presence of AMG. The results indicated that the isolated AMG, with a purity that exceeded 98%, had minimal inhibitory concentrations in the range of 4.6-9.2 μmol/L for the three strains tested. Interestingly, the MSSA strains were more sensitive to AMG than the MRSA strain. Minimal bactericidal concentrations were 2-fold higher than the minimal inhibitory concentration values for the three strains, indicating that AMG was a bactericidal compound. AMG also prevented biofilm formation effectively, albeit that again the MRSA strain was the most resistant. Interestingly, biofilms of the MRSA strain contained protein as a main component of the extracellular matrix, whereas this was polysaccharide in the MSSA strains. This might relate to the resistance of the MRSA 252 strain to AMG. Assays using human red blood cells indicated that AMG caused significant membrane damage with 50% of cell lysis occurred at concentration of about 36 μmol/L. Our results provide evidence that the isolated AMG has inhibitory activity against biofilm formation by S. aureus, including MRSA. Thus, isolated AMG proposes a high potential to develop a novel phytopharmaceutical for the treatment of MRSA.


Colon cancer survival with herbal medicine and vitamins combined with standard therapy in a whole-systems approach: ten-year follow-up data analyzed with marginal structural models and propensity score methods.
Although localized colon cancer is often successfully treated with surgery, advanced disease requires aggressive systemic therapy that has lower effectiveness. Approximately 30% to 75% of patients with colon cancer use complementary and alternative medicine (CAM), but there is limited formal evidence of survival efficacy. In a consecutive case series with 10-year follow-up of all colon cancer patients (n = 193) presenting at a San Francisco Bay-Area center for Chinese medicine (Pine Street Clinic, San Anselmo, CA), the authors compared survival in patients choosing short-term treatment lasting the duration of chemotherapy/radiotherapy with those continuing long-term. To put these data into the context of treatment responses seen in conventional medical practice, they also compared survival with Pan-Asian medicine + vitamins (PAM+V) with that of concurrent external controls from Kaiser Permanente Northern California and California Cancer Registries. Kaplan-Meier, traditional Cox regression, and more modern methods were used for causal inference-namely, propensity score and marginal structural models (MSMs), which have not been used before in studies of cancer survival and Chinese herbal medicine. PAM+V combined with conventional therapy, compared with conventional therapy alone, reduced the risk of death in stage I by 95%, stage II by 64%, stage III by 29%, and stage IV by 75%. There was no significant difference between short-term and long-term PAM+V. Combining PAM+V with conventional therapy improved survival, compared with conventional therapy alone, suggesting that prospective trials combining PAM+V with conventional therapy are justified.


Analysis of the diagnostic and prognostic value of miR-9-5p in carotid artery stenosis.
More and more evidence shows that microRNAs (miRNAs) play an important role in the diagnosis and prognosis of human diseases. In this study, we investigated the diagnostic value of miR-9-5p for asymptomatic carotid artery stenosis (CAS) and its predictive value for future cerebrovascular events within 5 years. A total of 88 asymptomatic CAS patients and 86 healthy individuals were recruited. The expression level of serum miR-9-5p was determined by quantitative real-time polymerase chain reaction (qRT-PCR). The diagnostic value of miR-9-5p in CAS was assessed by a receiving operator characteristic (ROC) curve. The predictive value of miR-9-5p for the occurrence of cerebrovascular events was evaluated by the Kaplan-Meier method. The serum level of miR-9-5p was significantly decreased in asymptomatic CAS patients. ROC curve had an AUC value of 0.910, with the sensitivity and specificity of 80.7% and 87.2% at the cut-off value of 0.72, respectively. A total of 25 patients had cerebrovascular events during the 5-year follow-up, including 3 strokes and 22 transient ischemic attacks (TIAs). Kaplan-Meier survival analysis revealed that the low expression level of miR-9-5p was an independent factor closely related to the occurrence of cerebrovascular events. Serum miR-9-5p could be used as a new biomarker for the diagnosis of CAS, and the low expression of miR-9-5p is associated with poor prognosis.


Out-of-hospital cardiac arrest due to coronary heart disease: a comparison of survival before and after the introduction of defribrillators in ambulances.
To assess the actual impact on coronary mortality of equipping ambulances with defibrillators. Retrospective analysis of routine medical and legal records of all those who had a cardiac arrest attributed to coronary heart disease occurring outside hospital in a defined population before and after the introduction of Heartstart. City of Glasgow, North of the River Clyde, 1984 and 1990. 296 and 267 men and women aged 25-64 inclusive in 1984 and 1990 respectively who had a cardiac arrest outside hospital which was attributed to coronary heart disease (International Classification of Diseases codes 410-414, ninth revision). The impact on coronary mortality in 1990 of equipping ambulances with defibrillators concurred with the earlier prediction of less than 1% of all coronary deaths. The circumstances of cardiac arrest were largely unchanged; most occurred outside hospital in the victim's home and the principal witnesses were members of the victim's family. A call for help before cardiac arrest was made in very few cases and cardiopulmonary resuscitation was attempted by laypersons in less than a third of the deaths they witnessed. There was a significant increase in the number of cardiopulmonary resuscitation attempts made by ambulance crews (16% v 32%, P < 0.01). Ambulance crews, however, still attended less than half of all cases (44% and 47%). The impact of equipping ambulances with defibrillators will remain small unless strategies are introduced that focus on improving the public's response to coronary emergencies by calling for help promptly and initiating cardiopulmonary resuscitation before the arrival of the emergency services.


Glycated hemoglobin as an indicator of social environmental stress among indigenous versus westernized populations.
This study assessed whether glycated hemoglobin concentration, an indicator of psychogenic stress, differs between indigenous populations and non-indigenous reference groups. Multivariate and stratified analyses were undertaken of cross-sectional data from multi-center community-based diabetes diagnostic and risk factor screening initiatives in Canada and Australia. Population groups were Australian Aborigines (n = 116), Torres Strait Islanders (n = 156), Native Canadians (n = 155), Greek migrants to Australia (n = 117), and Caucasian Australians (n = 67). Measurements included fasting glycated hemoglobin (HbA(1c)) concentration, fasting and 2-h post-load glucose concentrations, body mass index, waist-to-hip ratio, and demographic variables. Mean HbA(1c) concentrations were greater for indigenous groups than for Greek migrants and Caucasian Australians (P < 0. 0001). The covariate adjusted indigenous versus non-indigenous difference (95% CI) was 0.90 (0.58-1.22) percentage units, 18.2% higher for indigenous people. Stratified analyses indicated greater HbA(1c) for indigenous than for non-indigenous persons with normoglycemia (P = 0.009), impaired glucose tolerance (P = 0.097), and diabetes (P < 0.0001). HbA(1c) concentrations are greater for indigenous than for non-indigenous groups. Social changes, low control, and living conditions associated with westernization may be inherently stressful at the biological level for indigenous populations in westernized countries.


The behavior of lipid debris left on cell surfaces from microbubble based ultrasound molecular imaging.
Lipid monolayer coated microbubbles are currently being developed to identify vascular regions that express certain surface proteins as part of the new technique of ultrasound molecular imaging. The microbubbles are functionalized with targeting ligands which bind to the desired cells holding the microbubbles in place as the remaining unbound microbubbles are eliminated from circulation. Subsequent scanning with ultrasound can detect the highly reflectant microbubbles that are left behind. The ultrasound scanning and detection process results in the destruction of the microbubble, creating lipid fragments from the monolayer. Here we demonstrate that microbubbles targeted to 4T1 murine breast cancer cells and human umbilical cord endothelial cells leave behind adhered fragments of the lipid monolayer after exposure to ultrasound with peak negative pressures of 0.18 and 0.8MPa. Most of the observed fragments were large enough to be resistant to receptor mediated endocytosis. The fragments were not observed to incorporate into the lipid membrane of the cell over a period of 96min. They were not observed to break into smaller pieces or significantly change shape but they were observed to undergo translation and rotation across the cell surface as the cells migrated over the substrate. These large fragments will apparently remain on the surface of the targeted cells for significant periods of time and need to be considered for their potential effects on blood flow through the microcapillaries and potential for immune system recognition.


Genome-wide transcriptional response of primary alveolar macrophages following infection with porcine reproductive and respiratory syndrome virus.
Porcine reproductive and respiratory syndrome is a major cause of economic loss for the swine industry worldwide. Porcine reproductive and respiratory syndrome virus (PRRSV) triggers weak and atypical innate immune responses, but key genes and mechanisms by which the virus interferes with the host innate immunity have not yet been elucidated. In this study, genes that control the response of the main target of PRRSV, porcine alveolar macrophages (PAMs), were profiled in vitro with a time-course experiment spanning the first round of virus replication. PAMs were obtained from six piglets and challenged with the Lelystad PRRSV strain, and gene expression was investigated using Affymetrix microarrays and real-time PCR. Of the 1409 differentially expressed transcripts identified by analysis of variance, two, five, 25, 16 and 100 differed from controls by a minimum of 1.5-fold at 1, 3, 6, 9 and 12 h post-infection (p.i.), respectively. A PRRSV infection effect was detectable between 3 and 6 h p.i., and was characterized by a consistent downregulation of gene expression, followed by the start of the host innate immune response at 9 h p.i. The expression of beta interferon 1 (IFN-beta), but not of IFN-alpha, was strongly upregulated, whilst few genes commonly expressed in response to viral infections and/or induced by interferons were found to be differentially expressed. A predominance of anti-apoptotic transcripts (e.g. interleukin-10), a shift towards a T-helper cell type 2 response and a weak upregulation of tumour necrosis factor-alpha expression were observed within 12 h p.i., reinforcing the hypotheses that PRRSV has developed sophisticated mechanisms to escape the host defence.


Nonprescription antibiotic therapy: cultural models on both sides of the counter and both sides of the border.
Antibiotic resistance is a global public health threat exacerbated by medically unwarranted or improper antibiotic use. Pharmacy counters at the U.S.-Mexico border provide an example of where lay decisions to use antibiotics in ways considered "risky" may be initiated and negotiated. We test how cultural and public health knowledge of antibiotics is distributed among pharmacy staff, local Mexican clients, and U.S. medical tourists in the bordertown of Nogales using a cultural consensus tool. We find that shared cultural models across these groups include public health statements; however, other shared statements are likely to reinforce antibiotic sales at pharmacy counters by those on both sides of the purchase as economic, rather than therapeutic, encounters. From a public health perspective, border pharmacy counters are not a location where increased "safe" knowledge about antibiotic use is being transmitted. However, we do find a positive relationship between "safe" knowledge and reductions in risky behavior.


Type 2 diabetes and congenital hyperinsulinism cause DNA double-strand breaks and p53 activity in β cells.
β cell failure in type 2 diabetes (T2D) is associated with hyperglycemia, but the mechanisms are not fully understood. Congenital hyperinsulinism caused by glucokinase mutations (GCK-CHI) is associated with β cell replication and apoptosis. Here, we show that genetic activation of β cell glucokinase, initially triggering replication, causes apoptosis associated with DNA double-strand breaks and activation of the tumor suppressor p53. ATP-sensitive potassium channels (KATP channels) and calcineurin mediate this toxic effect. Toxicity of long-term glucokinase overactivity was confirmed by finding late-onset diabetes in older members of a GCK-CHI family. Glucagon-like peptide-1 (GLP-1) mimetic treatment or p53 deletion rescues β cells from glucokinase-induced death, but only GLP-1 analog rescues β cell function. DNA damage and p53 activity in T2D suggest shared mechanisms of β cell failure in hyperglycemia and CHI. Our results reveal membrane depolarization via KATP channels, calcineurin signaling, DNA breaks, and p53 as determinants of β cell glucotoxicity and suggest pharmacological approaches to enhance β cell survival in diabetes.


Attenuated Francisella novicida transposon mutants protect mice against wild-type challenge.
Francisella tularensis is the bacterial pathogen that causes tularemia in humans and a number of animals. To date, there is no approved vaccine for this widespread and life-threatening disease. The goal of this study was to identify F. tularensis mutants that can be used in the development of a live attenuated vaccine. We screened F. novicida transposon mutants to identify mutants that exhibited reduced growth in mouse macrophages, as these cells are the preferred host cells of Francisella and an essential component of the innate immune system. This approach yielded 16 F. novicida mutants that were 100-fold more attenuated for virulence in a mouse model than the wild-type parental strain. These mutants were then tested to determine their abilities to protect mice against challenge with high doses of wild-type bacteria. Five of the 16 attenuated mutants (with mutations corresponding to dsbB, FTT0742, pdpB, fumA, and carB in the F. tularensis SCHU S4 strain) provided mice with protection against challenge with high doses (>8 x 10(5) CFU) of wild-type F. novicida. We believe that these findings will be of use in the design of a vaccine against tularemia.


Excluding personality disorders from the insanity defense--a follow-up study.
Examining the effects of Oregon's statutory reform excluding personality disordered individuals from the insanity defense, we previously identified a study sample of insanity acquittees, each of whom was given a primary diagnosis of a personality disorder during subsequent evaluation at the state hospital. In the present study we explore the relationship between that diagnosis and the pretrial psychiatric diagnosis presented to the trial court. By reading the forensic mental health evaluations used at trial we found that 50 percent of our study sample of 34 personality disordered patients were diagnosed with psychotic disorders, affective disorders, retardation, and organic brain disorders. In addition to investigating the diagnosis offered as evidence at trial, we performed assessments of 38 mental health reports using published standards for forensic evaluation reports. We found compliance rates in the various categories ranged from 8 to 84 percent with a mean of 45 percent. We question the value of the mental health input to these trials, and believe that the data tend to validate past aspersions of forensic practice.


Single session real-time fMRI neurofeedback has a lasting impact on cognitive behavioral therapy strategies.
To benefit from cognitive behavioral therapy (CBT), individuals must not only learn new skills but also strategically implement them outside of session. Here, we tested a novel technique for personalizing CBT skills and facilitating their generalization to daily life. We hypothesized that showing participants the impact of specific CBT strategies on their own brain function using real-time functional magnetic imaging (rt-fMRI) neurofeedback would increase their metacognitive awareness, help them identify effective strategies, and motivate real-world use. In a within-subjects design, participants who had completed a clinical trial of a standardized course of CBT created a personal repertoire of negative autobiographical stimuli and mood regulation strategies. From each participant's repertoire, a set of experimental and control strategies were identified; only experimental strategies were practiced in the scanner. During the rt-fMRI neurofeedback session, participants used negative stimuli and strategies from their repertoire to manipulate activation in the anterior cingulate cortex, a region implicated in emotional distress. The primary outcome measures were changes in participant ratings of strategy difficulty, efficacy, and frequency of use. As predicted, ratings for unscanned control strategies were stable across observations, whereas ratings for experimental strategies changed after neurofeedback. At follow-up one month after the session, efficacy and frequency ratings for scanned strategies were predicted by neurofeedback during the rt-fMRI session. These results suggest that rt-fMRI neurofeedback created a salient and durable learning experience for patients, extending beyond the scan session to guide and motivate CBT skill use weeks later. This metacognitive approach to neurofeedback offers a promising model for increasing clinical benefits from cognitive behavioral therapy by personalizing skills and facilitating generalization.


Natural history of malignant bone disease in hepatocellular carcinoma: final results of a multicenter bone metastasis survey.
Bone is an uncommon site of metastasis in patients with advanced hepatocellular carcinoma (HCC). Therefore, there are few studies concerning the natural history of bone metastasis in patients with HCC. Data on clinicopathology, survival, skeletal-related events (SREs), and bone-directed therapies for 211 deceased HCC patients with evidence of bone metastasis were statistically analyzed. The median age was 70 years; 172 patients were male (81.5%). The median overall survival was 19 months. The median time to the onset of bone metastasis was 13 months (22.2% at HCC diagnosis); 64.9% patients had multiple bone metastases. Spine was the most common site of bone metastasis (59.7%). Most of these lesions were osteolytic (82.4%); 88.5% of them were treated with zoledronic acid. At multivariate analysis, only the Child Score was significantly correlated with a shorter time to diagnosis of bone metastases (p = 0.001, HR = 1.819). The median survival from bone metastasis was 7 months. At multivariate analysis, HCC etiology (p = 0.005), ECOG performance status (p = 0.002) and treatment with bisphosphonate (p = 0.024) were associated with shorter survival after bone disease occurrence. The site of bone metastasis but not the number of bone lesions was associated with the survival from first skeletal related event (SRE) (p = 0.021) and OS (p = 0.001). This study provides a significant improvement in the understanding the natural history of skeletal disease in HCC patients. An early and appropriate management of these patients is dramatically needed in order to avoid subsequent worsening of their quality of life.


An item response theory analysis of the Executive Interview and development of the EXIT8: A Project FRONTIER Study.
The Executive Interview (EXIT25) is an effective measure of executive dysfunction, but may be inefficient due to the time it takes to complete 25 interview-based items. The current study aimed to examine psychometric properties of the EXIT25, with a specific focus on determining whether a briefer version of the measure could comprehensively assess executive dysfunction. The current study applied a graded response model (a type of item response theory model for polytomous categorical data) to identify items that were most closely related to the underlying construct of executive functioning and best discriminated between varying levels of executive functioning. Participants were 660 adults ages 40 to 96 years living in West Texas, who were recruited through an ongoing epidemiological study of rural health and aging, called Project FRONTIER. The EXIT25 was the primary measure examined. Participants also completed the Trail Making Test and Controlled Oral Word Association Test, among other measures, to examine the convergent validity of a brief form of the EXIT25. Eight items were identified that provided the majority of the information about the underlying construct of executive functioning; total scores on these items were associated with total scores on other measures of executive functioning and were able to differentiate between cognitively healthy, mildly cognitively impaired, and demented participants. In addition, cutoff scores were recommended based on sensitivity and specificity of scores. A brief, eight-item version of the EXIT25 may be an effective and efficient screening for executive dysfunction among older adults.


The halo effect: suppression of pink bollworm on non-Bt cotton by Bt cotton in China.
In some previously reported cases, transgenic crops producing insecticidal proteins from Bacillus thuringiensis (Bt) have suppressed insect pests not only in fields planted with such crops, but also regionally on host plants that do not produce Bt toxins. Here we used 16 years of field data to determine if Bt cotton caused this "halo effect" against pink bollworm (Pectinophora gossypiella) in six provinces of the Yangtze River Valley of China. In this region, the percentage of cotton hectares planted with Bt cotton increased from 9% in 2000 to 94% in 2009 and 2010. We found that Bt cotton significantly decreased the population density of pink bollworm on non-Bt cotton, with net decreases of 91% for eggs and 95% for larvae on non-Bt cotton after 11 years of Bt cotton use. Insecticide sprays targeting pink bollworm and cotton bollworm (Helicoverpa armigera) decreased by 69%. Previously reported evidence of the early stages of evolution of pink bollworm resistance to Bt cotton in China has raised concerns that if unchecked, such resistance could eventually diminish or eliminate the benefits of Bt cotton. The results reported here suggest that it might be possible to find a percentage of Bt cotton lower than the current level that causes sufficient regional pest suppression and reduces the risk of resistance.