Expert Search Strategies: The Information Retrieval Practices of Healthcare Information Professionals.
Healthcare information professionals play a key role in closing the knowledge gap between medical research and clinical practice. Their work involves meticulous searching of literature databases using complex search strategies that can consist of hundreds of keywords, operators, and ontology terms. This process is prone to error and can lead to inefficiency and bias if performed incorrectly. The aim of this study was to investigate the search behavior of healthcare information professionals, uncovering their needs, goals, and requirements for information retrieval systems. A survey was distributed to healthcare information professionals via professional association email discussion lists. It investigated the search tasks they undertake, their techniques for search strategy formulation, their approaches to evaluating search results, and their preferred functionality for searching library-style databases. The popular literature search system PubMed was then evaluated to determine the extent to which their needs were met. The 107 respondents indicated that their information retrieval process relied on the use of complex, repeatable, and transparent search strategies. On average it took 60 minutes to formulate a search strategy, with a search task taking 4 hours and consisting of 15 strategy lines. Respondents reviewed a median of 175 results per search task, far more than they would ideally like (100). The most desired features of a search system were merging search queries and combining search results. Healthcare information professionals routinely address some of the most challenging information retrieval problems of any profession. However, their needs are not fully supported by current literature search systems and there is demand for improved functionality, in particular regarding the development and management of search strategies.


[Value of supine thoracic radiography in the diagnosis and quantification of pleural effusions: comparison with sonography].
The purpose of this prospective study was to verify the value of the supine chest radiograph with regard to the recognition and quantification of pleural effusions. The findings of supine chest radiographs were compared to those of chest sonography in a total of 320 pleural cavities. The correct identification and quantification of a pleural effusion was possible by supine chest radiograph alone in only 55% of cases (176/320). In 20% (64/320) supine chest x-ray was falsely negative and in 15% (48/320) falsely positive. In 10% an existing effusion was in fact recognised on supine chest x-ray; however the amount of effusion was wrongly estimated.


What do Key Stakeholders Think About HIV Self-Testing in Canada? Results from a Cross-Sectional Survey.
Human immunodeficiency virus (HIV) self-testing presents an empowering alternative to facility-based testing for reaching undiagnosed HIV infected individuals, but is not currently available in Canada. We surveyed stakeholders (clinical providers, public health professionals, researchers) engaged in HIV testing initiatives nationwide to identify the concerns, opportunities and challenges to implementing HIV self-testing in Canada. An online cross-sectional survey was disseminated by the Canadian Institutes of Health Research Centre for REACH 2.0 National HIV & sexually transmitted and blood borne infections working group to stakeholders nationwide, with a target sample size of 200. Quantitative and qualitative data were analyzed using a mixed-methods, respondent-informed approach, to inform subsequent HIV self-testing in a country where self-testing is not yet accessible. A total of 183 responses were received. A majority (70.7%) (128/181) felt that self-testing was a necessary investment to reach the undiagnosed. 64.6% (117/181) felt that self-tests should be made available to their clients and 71.5% (128/179) of respondents agreed that self-test instructions required improvements. However, 50% (90/180) felt that self-testing will pose an economic challenge to current HIV testing models. Regardless, 21% urged for timely action and availability of HIV self-tests. Thematic analyses reflected the following concerns: (a) need for affordable self-tests, (b) need for expedited, customized, and accessible linkages to counselling, (c) concern for patients to cope with positive self-test results, (d) accuracy of self-tests to detect acute HIV and (e) liability in the context of non-disclosure. Stakeholders agreed to the provision of an option of HIV self-testing to reach the undiagnosed individuals. Concerns regarding costs and accuracy of self-tests, expedited linkages to counselling, and integration of self-test within prevailing HIV testing models, will need to be addressed before their widespread implementation.


XMam1, the Xenopus homologue of mastermind, is essential to primary neurogenesis in Xenopus laevis embryos.
Notch signaling is involved in cell fate determination and is evolutionally highly conserved in vertebrates and invertebrates. Mastermind is a nuclear protein which participates in Notch signaling and is involved in direct transactivation of target genes. Here we analyzed the expression and the function of Xenopus mastermind1 (XMam1) in the process of primary neurogenesis. XMam1 is 3,425 bp and encodes 1,139 amino acids. Overall, Mastermind proteins consist of a basic domain, two acidic domains and a glutamine-rich domain, which are highly conserved among species. The ubiquitous expression of XMam1 was observed in both maternal and zygotic stages. Whole-mount in situ hybridization showed that XMam1 mRNA was present in the ectoderm by the gastrula stage and localized at the anterior neural region in the neurula stage. Thereafter, XMam1 expression was restricted to the eye and otic vesicle in the tailbud-stage embryo. XMaml overexpression caused the repression of primary neural formation. The truncated form of XMam1 (lacking the C-terminus of XMam1; XMam1deltaC) led to excess formation of primary neurons. Furthermore, XMam1deltaC strongly repressed XESR-1 transactivation. These results show that XMaml is involved in primary neurogenesis by way of Notch signaling and is an essential component for transactivation of XESR-1 in Xenopus laevis embryos.


Are ICD recipients able to foresee if they want to withdraw therapy or deactivate defibrillator shocks?
Expert consensus statements on management of implantable cardioverter defibrillators (ICDs) emphasize the importance of having discussions about deactivation before and after implantation. These statements were developed with limited patient input. The purpose of this study was to identify the factors associated with patients' experiences of end-of-life discussions, attitudes towards such discussions, and attitudes towards withdrawal of therapy (i.e., generator replacement and deactivation) at end-of-life, in a large national cohort of ICD-recipients. We enrolled 3067 ICD-patients, administrating the End-of-Life-ICD-Questionnaire. Most (86%) had not discussed ICD-deactivation with their physician. Most (69%) thought discussions were best at end-of-life, but 40% stated that they never wanted the physician to initiate a discussion. Those unwilling to discuss deactivation were younger, had experienced battery replacement, had a longer time since implantation, and had better quality-of-life. Those with psychological morbidity were more likely to desire a discussion about deactivation. Many patients (39%) were unable to foresee what to decide about deactivation in an anticipated terminal condition. Women, those without depression, and those with worse ICD-related experiences were more indecisive about withdrawal of therapy. Irrespective of shock experiences, those who could take a stand regarding deactivation chose to keep shock therapies active in many cases (39%). Despite consensus statements recommending discussions about ICD-deactivation at the end-of-life, such discussion usually do not occur. There is substantial ambivalence and indecisiveness on the part of most ICD-patients in this nationwide survey about having these discussions and about expressing desires about deactivation in an anticipated end-of-life situation.


[Experience in using hair as the object of study in forensic medical molecular genetic expertise].
It is possible to overcome the difficulties in expert evaluation of hairs by using molecular genetic methods of analysis at the level of DNA. We assessed the possibility of using hair for forensic genetic identification of relation and personality identification. The efficacy of using just individual hairs in analysis has been demonstrated. The stability of results was evaluated and the possible artefacts caused by low content of DNA and its possible degradation detected and corrected.


Relative influence of genetics and shared environment on child mental health symptoms depends on comorbidity.
Comorbidity among childhood mental health symptoms is common in clinical and community samples and should be accounted for when investigating etiology. We therefore aimed to uncover latent classes of mental health symptoms in middle childhood in a community sample, and to determine the latent genetic and environmental influences on those classes. The sample comprised representative cohorts of twins. A questionnaire-based assessment of mental health symptoms was used in latent class analyses. Data on 3223 twins (1578 boys and 1645 girls) with a mean age of 7.5 years were analyzed. The sample was predominantly non-Hispanic Caucasian (92.1%). Latent class models delineated groups of children according to symptom profiles--not necessarily clinical groups but groups representing the general population, most with scores in the normative range. The best-fitting models suggested 9 classes for both girls and boys. Eight of the classes were very similar across sexes; these classes ranged from a "Low Symptom" class to a "Moderately Internalizing & Severely Externalizing" class. In addition, a "Moderately Anxious" class was identified for girls but not boys, and a "Severely Impulsive & Inattentive" class was identified for boys but not girls. Sex-combined analyses implicated moderate genetic influences for all classes. Shared environmental influences were moderate for the "Low Symptom" and "Moderately Internalizing & Severely Externalizing" classes, and small to zero for other classes. We conclude that symptom classes are largely similar across sexes in middle childhood. Heritability was moderate for all classes, but shared environment played a greater role for classes in which no one type of symptom predominated.


In silico analysis of a novel causative mutation in Cadherin23 gene identified in an Omani family with hearing loss.
Hereditary hearing loss is a heterogeneous group of complex disorders with an overall incidence of one in every 500 newborns presented as syndromic and non-syndromic forms. Cadherin-related 23 (CDH23) is one of the listed deafness causative genes. It is found to be expressed in the stereocilia of hair cells and in the retina photoreceptor cells. Defective CDH23 have been associated mostly with prelingual severe-to-profound sensorineural hearing loss (SNHL) in either syndromic (USH1D) or non-syndromic SNHL (DFNB12) deafness. The purpose of this study was to identify causative mutations in an Omani family diagnosed with severe-profound sensorineural hearing loss by whole exome sequencing technique and analyzing the detected variant in silico for pathogenicity using several in silico mutation prediction software. A novel homozygous missense variant, c.A7436C (p. D2479A), in exon 53 of CDH23 was detected in the family while the control samples were all negative for the detected variant. In silico mutation prediction analysis showed the novel substituted D2479A to be deleterious and protein destabilizing mutation at a conserved site on CDH23 protein. In silico mutation prediction analysis might be used as a useful molecular diagnostic tool benefiting both genetic counseling and mutation verification. The aspartic acid 2479 alanine missense substitution might be the main disease-causing mutation that damages CDH23 function and could be used as a genetic hearing loss marker for this particular Omani family.


Nasoseptal flap reconstruction of pediatric sellar defects: a radiographic feasibility study and case series.
In this study, we used computed tomography measurements to investigate the feasibility of nasoseptal flap reconstruction of sellar defects in children, and we reviewed our institutional experience with the procedure. Cross-sectional and case series. Pediatric tertiary care facility. We obtained 10 normal maxillofacial scans for each year of age from birth to 18. Computer-assisted nasal and skull-base measurements were performed. Patients with incomplete pneumatization were excluded from analysis. Reconstruction was presumed feasible if the ratio of nasoseptal flap length to associated sellar defect length was greater than 1. Chart review identified surgical patients. Of 190 scans, 125 had complete pneumatization. Of these, 120 (96%) displayed a ratio of nasoseptal flap length to sellar defect length greater than 1, suggesting that reconstruction would be feasible. Mean ratio of flap length to defect length for all subjects was 1.47 (SD 0.33; 95% CI, 1.41-1.53). Only 5 of 125 patients (4%) had a ratio less than 1; the median age for these patients was 15 years, which is older than the median age of 12 years for subjects with a ratio greater than 1 (P = .02). An inverse relationship was identified between age and ratio of flap length to defect length (r = -0.49, P < .001). Case series identified 6 children, ages 5 to 17; flap length was never described as a limitation. Nasoseptal flap length is not a limiting factor for reconstruction of pediatric sellar defects. When compared with older patients, younger patients tend to have greater nasoseptal flap length relative to sellar defect length.


[The structure of occupational morbidity in electric welders].
Among 6 nosologic entities of occupational diseases diagnosed in 323 electric welders of Moscow suburb, the most frequent one was pneumoconiosis caused by welding aerosol, then neurosensory deafness, chronic toxic dust bronchitis and chronic manganese intoxication. Malignancies located especially in larynx and lungs appeared important among concurrent diseases that could be considered occupational.


Nematode pyruvate dehydrogenase kinases: role of the C-terminus in binding to the dihydrolipoyl transacetylase core of the pyruvate dehydrogenase complex.
Pyruvate dehydrogenase kinases (PDKs) from the anaerobic parasitic nematode Ascaris suum and the free-living nematode Caenorhabditis elegans were functionally expressed with hexahistidine tags at their N-termini and purified to apparent homogeneity. Both recombinant PDKs (rPDKs) were dimers, were not autophosphorylated and exhibited similar specific activities with the A. suum pyruvate dehydrogenase (E1) as substrate. In addition, the activities of both PDKs were activated by incubation with PDK-depleted A. suum muscle pyruvate dehydrogenase complex (PDC) and were stimulated by NADH and acetyl-CoA. However, the recombinant A. suum PDK (rAPDK) required higher NADH/NAD+ ratios for half-maximal stimulation than the recombinant C. elegans PDK (rCPDK) or values reported for mammalian PDKs, as might be predicted by the more reduced microaerobic mitochondrial environment of the APDK. Limited tryptic digestion of both rPDKs yielded stable fragments truncated at the C-termini (trPDKs). The trPDKs retained their dimeric structure and exhibited substantial PDK activity with the A. suum E1 as substrate, but PDK activity was not activated by incubation with PDK-depleted A. suum PDC or stimulated by elevated NADH/NAD+ or acetyl-CoA/CoA ratios. Direct-binding assays demonstrated that increasing amounts of rCPDK bound to the A. suum PDK-depleted PDC. No additional rCPDK binding was observed at ratios greater than 20 mol of rCPDK/mol of PDC. In contrast, the truncated rCPDK (trCPDK) did not exhibit significant binding to the PDC. Similarly, a truncated form of rCPDK, rCPDK1-334, generated by mutagenesis, exhibited properties similar to those observed for trCPDK. These results suggest that the C-terminus of the PDK is not required for subunit association of the homodimer or catalysis, but instead seems to be involved in the binding of the PDKs to the dihydrolipoyl transacetylase core of the complex.


Healthcare provider relational quality is associated with better self-management and less treatment burden in people with multiple chronic conditions.
Having multiple chronic conditions (MCCs) can lead to appreciable treatment and self-management burden. Healthcare provider relational quality (HPRQ) - the communicative and interpersonal skill of the provider - may mitigate treatment burden and promote self-management. The objectives of this study were to 1) identify the associations between HPRQ, treatment burden, and psychosocial outcomes in adults with MCCs, and 2) determine if certain indicators of HPRQ are more strongly associated than others with these outcomes. This is a cross-sectional survey study of 332 people with MCCs. Patients completed a 7-item measure of HPRQ and measures of treatment and self-management burden, chronic condition distress, self-efficacy, provider satisfaction, medication adherence, and physical and mental health. Associations between HPRQ, treatment burden, and psychosocial outcomes were determined using correlational analyses and independent samples t-tests, which were repeated in item-level analyses to explore which indicators of HPRQ were most strongly associated with the outcomes. Most respondents (69%) were diagnosed with ≥3 chronic conditions. Better HPRQ was found to be associated with less treatment and self-management burden and better psychosocial outcomes (P<0.001), even after controlling for physical and mental health. Those reporting 100% adherence to prescribed medications had higher HPRQ scores than those reporting less than perfect adherence (P<0.001). HPRQ items showing the strongest associations with outcomes were "my healthcare provider spends enough time with me", "my healthcare provider listens carefully to me", and "I have trust in my healthcare provider". Good communication and interpersonal skills of healthcare providers may lessen feelings of treatment burden and empower patients to feel confident in their self-management. Patient trust in the provider is an important element of HPRQ. Educating healthcare providers about the importance of interpersonal and relational skills could lead to more patient-centered care.


Epidemiological and Clinical Characteristics of Young Patients With Breast Cancer in Brazil: A Retrospective Study.
Understanding the epidemiological profile of young patients with breast cancer (BC) is crucial for devising effective control strategies. However, data on Brazilian patients with BC, particularly concerning the association with pathogenic germline mutations, are limited. This retrospective cross-sectional study seeks to shed light on the epidemiological and genetic profiles of young patients with BC in Brazil, with a specific focus on the correlation between germline mutations and clinical outcomes. We analyzed medical records from two institutions in Minas Gerais, Brazil, focusing on young patients with BC diagnosed from January 2012 to December 2020. The collected variables included sociodemographic profile, tumor characteristics, mutational status, follow-up duration, and time from diagnosis to death. The findings highlight high rates of genetic mutations, more aggressive tumor characteristics, and the necessity for more radical surgical interventions in patients with mutations. In general, young patients with BC in Brazil present an unfavorable prognosis, emphasizing the need for early diagnosis and personalized treatment strategies.


Current approaches to treatments for schizophrenia spectrum disorders, part II: psychosocial interventions and patient-focused perspectives in psychiatric care.
Schizophrenia is a disabling psychiatric illness associated with disruptions in cognition, emotion, and psychosocial and occupational functioning. Increasing evidence shows that psychosocial interventions for people with schizophrenia, as an adjunct to medications or usual psychiatric care, can reduce psychotic symptoms and relapse and improve patients' long-term outcomes such as recovery, remission, and illness progression. This critical review of the literature was conducted to identify the common approaches to psychosocial interventions for people with schizophrenia. Treatment planning and outcomes were also explored and discussed to better understand the effects of these interventions in terms of person-focused perspectives such as their perceived quality of life and satisfaction and their acceptability and adherence to treatments or services received. We searched major health care databases such as EMBASE, MEDLINE, and PsycLIT and identified relevant literature in English from these databases. Their reference lists were screened, and studies were selected if they met the criteria of using a randomized controlled trial or systematic review design, giving a clear description of the interventions used, and having a study sample of people primarily diagnosed with schizophrenia. Five main approaches to psychosocial intervention had been used for the treatment of schizophrenia: cognitive therapy (cognitive behavioral and cognitive remediation therapy), psychoeducation, family intervention, social skills training, and assertive community treatment. Most of these five approaches applied to people with schizophrenia have demonstrated satisfactory levels of short- to medium-term clinical efficacy in terms of symptom control or reduction, level of functioning, and/or relapse rate. However, the comparative effects between these five approaches have not been well studied; thus, we are not able to clearly understand the superiority of any of these interventions. With the exception of patient relapse, the longer-term (eg, >2 years) effects of these approaches on most psychosocial outcomes are not well-established among these patients. Despite the fact that patients' perspectives on treatment and care have been increasingly concerned, not many studies have evaluated the effect of interventions on this perspective, and where they did, the findings were inconclusive. To conclude, current approaches to psychosocial interventions for schizophrenia have their strengths and weaknesses, particularly indicating limited evidence on long-term effects. To improve the longer-term outcomes of people with schizophrenia, future treatment strategies should focus on risk identification, early intervention, person-focused therapy, partnership with family caregivers, and the integration of evidence-based psychosocial interventions into existing services.


Impact on GABA systems in monogenetic developmental CNS disorders: Clues to symptomatic treatment.
Animal studies of several single-gene disorders demonstrate that reversing the molecular signaling deficits can result in substantial symptomatic improvements in function. Focusing on the ratio of excitation to inhibition as a potential pathophysiological hallmark, seven single-gene developmental CNS disorders are reviewed which are characterized by a striking dysregulation of neuronal inhibition. Deficits in inhibition and excessive inhibition are found. The examples of developmental disorders encompass Neurofibromatosis type 1, Fragile X syndrome, Rett syndrome, Dravet syndrome including autism-like behavior, NONO-mutation-induced intellectual disability, Succinic semialdehyde dehydrogenase deficiency and Congenital nystagmus due to FRMD7 mutations. The phenotype/genotype correlations observed in animal models point to potential treatment options and will continue to inspire clinical research. Three drugs are presently in clinical trials: acamprosate and ganoxolon for Fragile X syndrome and SGS-742 for SSADH deficiency. This article is part of the "Special Issue Dedicated to Norman G. Bowery".


Biomaterials as vaccine adjuvants.
Biomaterials are promising candidate adjuvants to enhance vaccine efficacy. Through adjuvant design, we can broaden the use of vaccines to diseases such as AIDS, malaria, and cancer. This review addresses the fundamentals of vaccine and adjuvant function in order to determine guidelines for adjuvant design, including aspects of the vaccine such as disease target, antigen formulation, and delivery route. An ideal biomaterial adjuvant will perform three functions. (1) It will deliver the antigen selectively to dendritic cells. This has been accomplished through release of chemokines or cytokines, use of antidendritic cell antibodies, and even through particle size selection. (2) It will activate the dendritic cells, improving antigen presentation. Biomaterials themselves have been shown to activate innate immunity, but specific innate-activating ligands have also been included in adjuvant formulations. Finally, (3) it will release the antigen appropriately into the dendritic cell. Tuning release to be pH sensitive and engineering endosomal release are strategies that have been used. There is a real opportunity to rationally design better biomaterial adjuvants that will significantly expand and improve vaccine function.


Ectopic lignification in primary cellulose-deficient cell walls of maize cell suspension cultures.
Maize (Zea mays L.) suspension-cultured cells with up to 70% less cellulose were obtained by stepwise habituation to dichlobenil (DCB), a cellulose biosynthesis inhibitor. Cellulose deficiency was accompanied by marked changes in cell wall matrix polysaccharides and phenolics as revealed by Fourier transform infrared (FTIR) spectroscopy. Cell wall compositional analysis indicated that the cellulose-deficient cell walls showed an enhancement of highly branched and cross-linked arabinoxylans, as well as an increased content in ferulic acid, diferulates and p-coumaric acid, and the presence of a polymer that stained positive for phloroglucinol. In accordance with this, cellulose-deficient cell walls showed a fivefold increase in Klason-type lignin. Thioacidolysis/GC-MS analysis of cellulose-deficient cell walls indicated the presence of a lignin-like polymer with a Syringyl/Guaiacyl ratio of 1.45, which differed from the sensu stricto stress-related lignin that arose in response to short-term DCB-treatments. Gene expression analysis of these cells indicated an overexpression of genes specific for the biosynthesis of monolignol units of lignin. A study of stress signaling pathways revealed an overexpression of some of the jasmonate signaling pathway genes, which might trigger ectopic lignification in response to cell wall integrity disruptions. In summary, the structural plasticity of primary cell walls is proven, since a lignification process is possible in response to cellulose impoverishment.


S100A4 silencing suppresses proliferation, angiogenesis and invasion of thyroid cancer cells through downregulation of MMP-9 and VEGF.
It is well documented that S100A4 is upregulated in many cancers and plays a pivotal role in tumor proliferation, invasion, metastasis and angiogenesis. However, the precise role and mechanism S100A4 exerts in the thyroid cancer have not been fully elucidated to date. In the present study, we investigated the effect of S100A4 on proliferation, invasion, metastasis and angiogenesis in thyroid cancer cells. A plasmid construct was made that expressed full long S100A4 cDNA. The construct was stably transfected into BCPAP and ML-1 thyroid cancer cells (BCPAP/ S100A4 cDNA, ML-1/S100A4 cDNA). S100A4 siRNA was transiently transfected into the DRO cells (DRO/S100A4 siRNA). MMP-9 siRNA or VEGF siRNA was transiently transfected into the BCPAP/ S100A4 cDNA, ML-1/S100A4 cDNA cells (BCPAP/ S100A4 cDNA/VEGF siRNA, ML-1/S100A4 cDNA/ MMP-9 siRNA). We found that the down-regulation of S100A4 by small interfering RNA decreased cell invasion, metastasis, and angiogenesis by using chicken chorioallantoic membrane (CAM), whereas S100A4 overexpression by cDNA transfection led to increased tumor cell invasion, metastasis, and angiogenesis. Consistent with these results, we found that the down-regulation of S100A4 reduced VEGF and MMP-9 expression. Furthermore, Knockdown of MMP-9 by MMP-9 siRNA inhibited cell invasion and metastasis in the BCPAP/S100A4 cDNA, ML-1/S100A4 cDNA cells. Knockdown of VEGF by VEGF siRNA inhibited cell angiogenesis in the BCPAP/ S100A4 cDNA, ML-1/S100A4 cDNA cells.We also found that downregulation of S100A4 by small interfering RNA resulted in enhanced cell growth inhibition and apoptosis, and vice versa. Our data suggest S100A4 could be an effective approach for the regulation of proliferation, invasion and angiogenesis. Downregulation of S100A4 could inhibit angiogenesis, proliferation and invasion by regulating the expression of MMP-9 and VEGF. Our results provide evidence that the downregulation of S100A4 using RNAi technology may provide an effective tool for thyroid cancer therapy.


Molecular dynamics simulations reveal structural coordination of Ffh-FtsY heterodimer toward GTPase activation.
Two homologous GTPases (guanine-triphosphatases) in the signal recognition particle (SRP) and its receptor (SR) use their cumulative energy during GTP (guanine-triphosphate) hydrolysis to control the co-translational protein targeting process. Distinct from classical GTPases, which rely on external factors to hydrolyze GTP, SRP GTPases stimulate one another's activity in a self-sufficient manner upon SRP-SR complex association. Although both ground-state and putative transition-state GTP analogs have been used to recapitulate the state of GTPase activation, the underlying mechanism of the activated state still remains elusive. In particular, several residues that were placed in pending positions have been shown to be important to GTP hydrolysis in biochemical studies. Here, we examined the stability and dynamics of three interaction networks involving these residues and discovered that they contribute to the GTPase activation via well-tuned conformational changes. The crystallographically identified pending residues Ffh:R191/FtsY:R195 undergo extensive conformational rearrangements to form persisted interactions with FtsY:E284/Ffh:E274, explaining the biochemically observed defective effect of R191 mutant to the activation of both GTPases. In addition, the side chain of FtsY:R142, one of the most important catalytic residues, rotates to an extended conformation that could more efficiently maintain the electrostatic balance for GTP hydrolysis. Finally, the invariant residues Ffh:G190 and FtsY:G194, instead of the supposed auxiliary water molecules, are proposed to stabilize the nucleophilic waters during GTPase activation. In complementary to experimental observations, these findings suggest a more favorable interaction model for SRP GTPase activation and would thus benefit to our understanding of how SRP GTPases regulate the protein targeting pathway.


Impact of deoxynivalenol (DON) contaminated feed on intestinal integrity and immune response in swine.
This study was performed to characterize the influence of consuming DON naturally contaminated feeds on pig's intestinal immune defenses, antibody response and cellular immunity. Sixteen 4-week-old piglets were randomly allocated to two dietary treatments: control diet or diet contaminated with 3.5 mg DON/kg. At days 7 and 21, animals were immunized with ovalbumin (OVA). On day 42, intestinal samples were collected for measurement of gene expression involved in immune response, oxidative status and barrier function. Primary IgG antibody response to OVA was increased in pigs fed DON diet compared to control animals. In the ileum of pigs fed DON diet, claudin, occludin, and vimentin genes involved in integrity and barrier function were down-regulated compared to controls. Results also revealed that expression of two chemokines (IL-8, CXCL10), interferon-γ, and major antioxidant glutathione peroxidase 2 (GPX-2) were up-regulated whereas expression of genes encoding enzymatic antioxidants including GPX-3, GPX-4 and superoxide dismutase 3 (SOD-3) were down-regulated in pigs fed DON-contaminated diet. These results strongly suggest that ingestion of DON naturally contaminated feed impaired intestinal barrier and immunological functions by modulating expression of genes coding for proteins involved in tight junctions, tissue remodelling, inflammatory reaction, oxidative stress reaction and immune response.


Spidroin profiling of cribellate spiders provides insight into the evolution of spider prey capture strategies.
Orb-weaving spiders have two main methods of prey capture: cribellate spiders use dry, sticky capture threads, and ecribellate spiders use viscid glue droplets. Predation behaviour is a major evolutionary driving force, and it is important on spider phylogeny whether the cribellate and ecribellate spiders each evolved the orb architecture independently or both strategies were derived from an ancient orb web. These hypotheses have been discussed based on behavioural and morphological characteristics, with little discussion on this subject from the perspective of molecular materials of orb web, since there is little information about cribellate spider-associated spidroin genes. Here, we present in detail a spidroin catalogue of six uloborid species of cribellate orb-weaving spiders, including cribellate and pseudoflagelliform spidroins, with transcriptome assembly complemented with long read sequencing, where silk composition is confirmed by proteomics. Comparative analysis across families (Araneidae and Uloboridae) shows that the gene architecture, repetitive domains, and amino acid frequencies of the orb web constituting silk proteins are similar among orb-weaving spiders regardless of the prey capture strategy. Notably, the fact that there is a difference only in the prey capture thread proteins strongly supports the monophyletic origin of the orb web.


Charge-reversal lipids, peptide-based lipids, and nucleoside-based lipids for gene delivery.
Twenty years after gene therapy was introduced in the clinic, advances in the technique continue to garner headlines as successes pique the interest of clinicians, researchers, and the public. Gene therapy's appeal stems from its potential to revolutionize modern medical therapeutics by offering solutions to myriad diseases through treatments tailored to a specific individual's genetic code. Both viral and non-viral vectors have been used in the clinic, but the low transfection efficiencies when non-viral vectors are used have lead to an increased focus on engineering new gene delivery vectors. To address the challenges facing non-viral or synthetic vectors, specifically lipid-based carriers, we have focused on three main themes throughout our research: (1) The release of the nucleic acid from the carrier will increase gene transfection. (2) The use of biologically inspired designs, such as DNA binding proteins, to create lipids with peptide-based headgroups will improve delivery. (3) Mimicking the natural binding patterns observed within DNA, by using lipids having a nucleoside headgroup, will produce unique supramolecular assembles with high transfection efficiencies. The results presented in this Account demonstrate that engineering the chemical components of the lipid vectors to enhance nucleic acid binding and release kinetics can improve the cellular uptake and transfection efficacy of nucleic acids. Specifically, our research has shown that the incorporation of a charge-reversal moiety to initiate a shift of the lipid from positive to negative net charge improves transfection. In addition, by varying the composition of the spacer (rigid, flexible, short, long, or aromatic) between the cationic headgroup and the hydrophobic chains, we can tailor lipids to interact with different nucleic acids (DNA, RNA, siRNA) and accordingly affect delivery, uptake outcomes, and transfection efficiency. The introduction of a peptide headgroup into the lipid provides a mechanism to affect the binding of the lipid to the nucleic acid, to influence the supramolecular lipoplex structure, and to enhance gene transfection activity. Lastly, we discuss the in vitro successes that we have had when using lipids possessing a nucleoside headgroup to create unique self-assembled structures and to deliver DNA to cells. In this Account, we state our hypotheses and design elements as well as describe the techniques that we have used in our research to provide readers with the tools to characterize and engineer new vectors.


LAPTM4B overexpression is a novel independent prognostic marker for metastatic ovarian tumors.
Metastatic ovarian tumors are a series of lethal carcinomas that almost always have bad prognosis. Their prognoses, however, vary depending on the primary tumor malignancies of each. It has been reported that LAPTM4B, a novel tumor-associated gene, might indicate a worse prognosis when it was overexpressed in other carcinomas. Therefore, the authors expected to investigate whether LAPTM4B overexpression is an independent prognostic marker in metastatic ovarian tumors. Immunohistochemistry was used to assess LAPTM4B expression in metastatic ovarian tumors from 102 patients. Subsequently, univariate and multivariate survival analyses with Cox regression were performed to determine the association between LAPTM4B expression and prognosis. To identify any differences in prognosis between the 2 groups of patients with differing primary malignancies, the log-rank test was used. The median overall and progression-free survival rates of patients with tumors of gastrointestinal tract origin were 0.97 and 0.51 years, respectively, and both were statistically significantly lower than those of patients with tumors of breast origin (P < 0.0001), which were 2.68 and 1.96 years, accordingly. Of 102 patients, 77 were classified as having a high expression of LAPTM4B, and LAPTM4B expression had a significant association with the prognosis of metastatic ovarian tumors (P < 0.01); no statistically significant interaction between LAPTM4B expression and primary malignancies was detected (P > 0.1). On the other hand, medians of overall survival and progression-free survival of patients with tumors of gastrointestinal tract origin were significantly lower than those of patients with tumors of breast origin (P < 0.0001). Patients with metastatic ovarian tumors of breast origin had significantly better prognosis than those with the disease from gastrointestinal tract primary malignancies. LAPTM4B overexpression might be an independent prognostic marker of metastatic ovarian tumors.


Improving hybrid rice breeding programs via stochastic simulations: number of parents, number of hybrids, tester update, and genomic prediction of hybrid performance.
Schemes that use genomic prediction outperform others, updating testers increases hybrid genetic gain, and larger population sizes tend to have higher genetic gain and less depletion of genetic variance One of the most common methods to improve hybrid performance is reciprocal recurrent selection (RRS). Genomic prediction (GP) can be used to increase genetic gain in RRS by reducing cycle length, but it is also possible to use GP to predict single-cross hybrid performance. The impact of the latter method on genetic gain has yet to be previously reported. Therefore, we compared via stochastic simulations various phenotypic and genomics-assisted RRS breeding schemes which used GP to predict hybrid performance rather than reducing cycle length, which allows minimal changes to traditional breeding schemes. We also compared three breeding sizes scenarios that varied the number of genotypes crossed within heterotic pools, the number of genotypes crossed between heterotic pools, the number of hybrids evaluated, and the number of genomic predicted hybrids. Our results demonstrated that schemes that used genomic prediction of hybrid performance outperformed the others for the average interpopulation hybrid population and the best hybrid performance. Furthermore, updating the testers increased hybrid genetic gain with phenotypic RRS. As expected, the largest breeding size tested had the highest rates of genetic improvement and the lowest decrease in additive genetic variance due to the drift. Therefore, this study demonstrates the usefulness of single-cross prediction, which may be easier to implement than rapid-cycling RRS and cyclical updating of testers. We also reiterate that larger population sizes tend to have higher genetic gain and less depletion of genetic variance.


Consensus on the current use of sorafenib for the treatment of hepatocellular carcinoma.
An expert panel was convened to reach a consensus on the current use of sorafenib in the treatment of hepatocellular carcinoma (HCC). A multinational, multidisciplinary group of experts objectively reviewed clinical data of sorafenib and considered clinical experience to develop statements summarizing our consensus on the current use of sorafenib. Sorafenib is the standard of care for Child-Pugh (CP) A patients with advanced HCC (i.e. not amenable to surgery or locoregional treatment). There is currently insufficient evidence to support the routine clinical use of sorafenib in CP B patients, but sorafenib can be offered as an option for those with compensated liver cirrhosis. Patients with stable performance status who have progressive disease during sorafenib treatment can be maintained on sorafenib, although there are no clear data supporting the continuation of sorafenib in these patients. The effectiveness of the adjuvant use of sorafenib in preventing recurrence after resection or local ablation, or in combination with transarterial chemoembolization, are being evaluated in clinical studies. In conclusion, sorafenib has extended treatment options for patients with HCC, and is now established as the standard of care for CP A patients with HCC not amenable to surgery or locoregional treatment. Based on its multikinase-inhibitor activity and proven efficacy in prolonging survival in HCC, broader use - including a subgroup of patients with CP B (clinically compensated cirrhosis) and earlier disease stages - might be expected dependent on the results of ongoing studies of safety and clinically relevant benefit in these patients.


Immunological Phenotyping of Mice with a Point Mutation in Cdk4.
Cyclin-dependent kinases (CDKs) play a crucial role in regulation of the mammalian cell cycle. CDK4 and CDK6 control the G1/S restriction checkpoint through their ability to associate with cyclin D proteins in response to growth factor signals. CDK4 deficiency in mice gives rise to a range of endocrine-specific phenotypes including diabetes, infertility, dwarfism, and atrophy of the anterior pituitary. Although CDK6 deficiency can cause thymic atrophy due to a block in the double-negative (DN) to double-positive (DP) stage of T cell development, there are no overt defects in immune cell development reported for CDK4-deficient mice. Here, we examined the impact of a novel N-ethyl-N-nitrosourea-induced point mutation in the gene encoding CDK4 on immune cell development. Mutant mice (Cdk4wnch/wnch) showed normal development and differentiation of major immune cell subsets in the thymus and spleen. Moreover, T cells from Cdk4wnch/wnch mice exhibited normal cytokine production in response to in vitro stimulation. However, analysis of the mixed bone marrow chimeras revealed that Cdk4wnch/wnch-derived T cell subsets and NK cells are at a competitive disadvantage compared to Cdk4+/+-derived cells in the thymus and periphery of recipients. These results suggest a possible role for the CDK4wnch mutation in the development of some immune cells, which only becomes apparent when the Cdk4wnch/wnch mutant cells are in direct competition with wild-type immune cells in the mixed bone marrow chimera.


Sources of top-down control in visual search.
Endogenous control of visual search can influence search guidance at the level of a supradimensional topographic saliency map [Wolfe, J. M. Guided Search 2.0: A revised model of visual search. Psychonomic Bulletin & Review, 1, 202-238, 1994], and modulate nonspatial mechanisms coding saliency in dimension-specific input modules [Müller, H. J., Reimann, B., & Krummenacher, J. Visual search for singleton feature targets across dimensions: Stimulus- and expectancy-driven effects in dimensional weighting. Journal of Experimental Psychology: Human Perception and Performance, 29, 1021-1035, 2003]. The current experiment used fMRI to dissociate these mechanisms in a singleton feature search task in which the likely target dimension (color or orientation) was semantically precued and target saliency in each dimension was varied parametrically. BOLD signal increases associated with increased demands for top-down guidance were observed within the fronto-parietal attention network and in the right anterior middle frontal gyrus. Decreasing requirements for top-down control led to BOLD signal increases in medial anterior prefrontal cortex, consistent with a gating mechanism in favor of stimulus-related processing [Burgess, P. W., Dumontheil, I., & Gilbert, S. J. The gateway hypothesis of rostral prefrontal cortex (area 10) function. Trends in Cognitive Sciences, 11, 290-298, 2007]. Another network of brain areas consisting of left lateral fronto-polar cortex, the left supramarginal gyrus, and the cerebellum, as well as a bilateral network consisting of the posterior orbital gyrus, the inferior frontal gyrus, and the pre-SMA were associated with top-down dimensional (re-) orienting. These data argue in favor of distinct endogenous control systems for visuospatial and dimension-based attentional processing. Finally, cue validity modulated saliency processing in the left temporo-parietal junction (TPJ), pointing to a crucial role of the left TPJ in integrating an endogenous dimension-based attention set with bottom-up saliency signals.


Inhibition of virus replication by RNA interference.
RNA interference (RNAi) is a sequence-specific gene-silencing mechanism in eukaryotes, which is believed to function as a defence against viruses and transposons. Since its discovery, RNAi has been developed into a widely used technique for generating genetic knock-outs and for studying gene function by reverse genetics. Additionally, inhibition of virus replication by means of induced RNAi has now been reported for numerous viruses, including several important human pathogens such as human immunodeficiency virus type 1, hepatitis C virus, hepatitis B virus, dengue virus, poliovirus and influenza virus A. In this review, we will summarize the current data on RNAi-mediated inhibition of virus replication and discuss the possibilities for the development of RNAi-based antiviral therapeutics.


Team behaviors in emergency care: a qualitative study using behavior analysis of what makes team work.
Teamwork has been suggested as a promising approach to improving care processes in emergency departments (ED). However, for teamwork to yield expected results, implementation must involve behavior changes. The aim of this study is to use behavior analysis to qualitatively examine how teamwork plays out in practice and to understand eventual discrepancies between planned and actual behaviors. The study was set in a Swedish university hospital ED during the initial phase of implementation of teamwork. The intervention focused on changing the environment and redesigning the work process to enable teamwork. Each team was responsible for entire care episodes, i.e. from patient arrival to discharge from the ED. Data was collected through 3 days of observations structured around an observation scheme. Behavior analysis was used to pinpoint key teamwork behaviors for consistent implementation of teamwork and to analyze the contingencies that decreased or increased the likelihood of these behaviors. We found a great discrepancy between the planned and the observed teamwork processes. 60% of the 44 team patients observed were handled solely by the appointed team members. Only 36% of the observed patient care processes started according to the description in the planned teamwork process, that is, with taking patient history together. Beside this behavior, meeting in a defined team room and communicating with team members were shown to be essential for the consistent implementation of teamwork. Factors that decreased the likelihood of these key behaviors included waiting for other team members or having trouble locating each other. Getting work done without delay and having an overview of the patient care process increased team behaviors. Moreover, explicit instructions on when team members should interact and communicate increased adherence to the planned process. This study illustrates how behavior analysis can be used to understand discrepancies between planned and observed behaviors. By examining the contextual conditions that may influence behaviors, improvements in implementation strategies can be suggested. Thereby, the adherence to a planned intervention can be improved, and/or revisions of the intervention be suggested.


Interventions in chronic pain management. 2. Diagnosis of cervical and thoracic pain syndromes.
This self-directed learning module highlights approaches to the investigation of common cervical and thoracic conditions. It is part of the chapter on chronic pain management in the Self-Directed Physiatric Education Program for practitioners in physical medicine and rehabilitation. The differential diagnosis of cervical and thoracic syndromes as well as cervicogenic headache is reviewed. The need for a comprehensive history and physical examination is emphasized. Indications for diagnostic tests including magnetic resonance imaging, computed tomography scan, bone scan, diskography, radiographs, diagnostic injections, and electrodiagnostic studies are discussed with the idea that testing should be performed and interpreted with the specific clinical presentation in mind. To discuss the differential diagnoses for and investigation of common cervical and thoracic conditions and cervicogenic headache.


Spontaneous resolution of primary hyperparathyroidism post-biopsy-related neck haematoma.
Spontaneous or fine-needle aspiration (FNA)-induced remission of primary hyperparathyroidism (PHPT) is an extremely rare phenomenon with variable outcomes. We report a 75-year-old Male who initially presented with left ureteric calculi and was found to have PHPT. Imaging studies including ultrasound neck, parathyroid sestamibi scan and computed tomography of thorax, abdomen, and pelvis failed to identify the culprit lesion and exploratory parathyroidectomy was planned. Before surgery, he underwent FNA for cytology of a right cold thyroid nodule which was complicated with a large neck haematoma and dysphagia. The cytology of the aspirated fluid was consistent with a benign cyst. One month after the procedure, serum calcium and phosphate normalised along with resolution of haematoma. He remained in biochemical remission at 1-year follow-up with the latest ultrasound of neck showing resolution of a large colloid nodule that was previously seen occupying the right thyroid lobe.


[The duration of the meiosis stages in fertilized ovocytes of the mussel Mytilus galloprovincialis Lam. at different temperatures].
Duration of meiosis steps in fertilized oocytes of mussel Mytilus galloprovincialis at three levels of development temperature optimum: 8.5, 12 and 17 degrees C. Absolute duration of meiosis stages increases with decreasing of water temperature. Relative duration of analogous stages doesn't depend on temperature conditions. Duration of first meiotic division determined by the time between maximal occurrences of consequent stages, at different temperatures is in two times greater than duration of second division.


Assessment of automated disease detection in diabetic retinopathy screening using two-field photography.
To assess the performance of automated disease detection in diabetic retinopathy screening using two field mydriatic photography. Images from 8,271 sequential patient screening episodes from a South London diabetic retinopathy screening service were processed by the Medalytix iGrading™ automated grading system. For each screening episode macular-centred and disc-centred images of both eyes were acquired and independently graded according to the English national grading scheme. Where discrepancies were found between the automated result and original manual grade, internal and external arbitration was used to determine the final study grades. Two versions of the software were used: one that detected microaneurysms alone, and one that detected blot haemorrhages and exudates in addition to microaneurysms. Results for each version were calculated once using both fields and once using the macula-centred field alone. Of the 8,271 episodes, 346 (4.2%) were considered unassessable. Referable disease was detected in 587 episodes (7.1%). The sensitivity of the automated system for detecting unassessable images ranged from 97.4% to 99.1% depending on configuration. The sensitivity of the automated system for referable episodes ranged from 98.3% to 99.3%. All the episodes that included proliferative or pre-proliferative retinopathy were detected by the automated system regardless of configuration (192/192, 95% confidence interval 98.0% to 100%). If implemented as the first step in grading, the automated system would have reduced the manual grading effort by between 2,183 and 3,147 patient episodes (26.4% to 38.1%). Automated grading can safely reduce the workload of manual grading using two field, mydriatic photography in a routine screening service.


TMEM120A contains a specific coenzyme A-binding site and might not mediate poking- or stretch-induced channel activities in cells.
TMEM120A, a member of the transmembrane protein 120 (TMEM120) family, has a pivotal function in adipocyte differentiation and metabolism, and may also contribute to sensing mechanical pain by functioning as an ion channel named TACAN. Here we report that expression of TMEM120A is not sufficient in mediating poking- or stretch-induced currents in cells and have solved cryo-electron microscopy (cryo-EM) structures of human TMEM120A (HsTMEM120A) in complex with an endogenous metabolic cofactor (coenzyme A, CoASH) and in the apo form. HsTMEM120A forms a symmetrical homodimer with each monomer containing an amino-terminal coiled-coil motif followed by a transmembrane domain with six membrane-spanning helices. Within the transmembrane domain, a CoASH molecule is hosted in a deep cavity and forms specific interactions with nearby amino acid residues. Mutation of a central tryptophan residue involved in binding CoASH dramatically reduced the binding affinity of HsTMEM120A with CoASH. HsTMEM120A exhibits distinct conformations at the states with or without CoASH bound. Our results suggest that TMEM120A may have alternative functional roles potentially involved in CoASH transport, sensing, or metabolism.


Monovalent cations differentially affect membrane surface properties and membrane curvature, as revealed by fluorescent probes and dynamic light scattering.
The effects of monovalent cations on the interfacial electrostatic potential (psi d), hydrodynamic shear boundary distance (ds), and membrane curvature were studied in large unilamellar phospholipid and galacto/sulfolipid liposomes containing different fractions of negatively charged lipids. The differential effects of alkali metal ions on psi d could be accurately determined at physiological surface charge densities with a surface-anchored fluorescent probe. Li+ and Na+ more effectively decrease psi d and exhibit higher association constants (Kas) than K+ and Cs+. These two groups of cations display qualitatively different perturbations of the interfacial structure. Combining Kas values with the electrokinetic (zeta) potentials yielded the respective ds values. At low ionic strength ds more substantially increases with Li+ or Na+ than with K+ or Cs+. Increasing surface charge density causes increased membrane curvature in the presence of K+ or Cs+, but this is largely prevented by Li+ or Na+. Membrane binding of the amphiphilic cation acridine orange decreases surface charge and membrane curvature more extensively than H3O+, Li+, and Na+. The differential interface-perturbing behavior of monovalent cations is discussed with regard to their different hydration tendencies that will modulate the extent and stability of the hydrogen-bond network along the charged membrane surface.


Tumor Mutational Burden, Toxicity, and Response of Immune Checkpoint Inhibitors Targeting PD(L)1, CTLA-4, and Combination: A Meta-regression Analysis.
Tumor mutational burden (TMB) has emerged as a potential predictive biomarker for clinical response to ICI therapy, but whether TMB also predicts toxicity remains unknown. We investigated the relationship between TMB, objective response rate (ORR), overall survival (OS), and toxicity for ICI therapy across multiple cancer types. We searched MEDLINE, PubMed, and ASCO/ESMO/AACR meetings for clinical trials of anti-PD(L)1, CTLA-4, or combination in 29 cancer types. We assessed ICI administered, responses (complete or partial response), median OS, OS HR, and grade 3/4 toxicity. We conducted a systematic review, meta-analysis and meta-regression using tumor level TMB data from Foundation Medicine. One hundred seventeen clinical trials, which included 12,450 patients treated with ICI therapy were analyzed. Meta-regression analysis revealed that TMB was significantly associated with ORR for anti-PD(L)1, anti-CTLA-4, and combination (P < 0.0001 for all), but not associated with toxicity in all treatment groups. OS data were unavailable for most studies included in our meta-analysis, and the relationship between TMB and OS in this subset was not significant (P = 0.26). In high TMB tumor types (≥10 mut/megabase) the improvement of ORR and increase in grade 3/4 toxicity with combination ICI therapy as compared with PD(L)1 monotherapy were 21.13% and 25.41%, respectively, as compared with 3.73% and 18.78% in low TMB tumor types (<10 mut/megabase). There is a positive association between TMB and clinical response with anti-PD(L)1, anti-CTLA-4, and combination ICIs, but no association between TMB and toxicity. These results imply a favorable risk/benefit ratio for ICIs in tumors with a higher TMB.


Genome-wide homozygosity mapping in families with leber congenital amaurosis identifies mutations in AIPL1 and RDH12 genes.
Leber congenital amaurosis (LCA) causes severe visual impairment and blindness very early in life. Mutant alleles of several genes acting in different pathways, of which all have critical roles for normal retinal function, were involved in LCA development. The purpose of this study was to use genome-wide genotyping to identify LCA-causing loci in two Turkish families. Genome-wide genotyping and haplotype analysis were performed for prioritization of candidate genes for mutation screening in families with LCA. Identified informative critical choromosomal regions obtained by homozygosity mapping from the families were searched for overlapping of any LCA causative genes. Corresponding clinical phenotypes of the patients with identified mutations were evaluated. In this study, two families were shown to be linked to two different LCA loci covering retinol dehydrogenase 12 (RDH12) and aryl-hydrocarbon-interacting protein-like1 (AIPL1) genes. Mutation screening revealed a novel p.Gln141* mutation in the AIPL1 gene and a previously described p.Thr49Met mutation in the RDH12 gene in a homozygous state. Our patients with the RDH12 mutation had the distinct macular coloboma sign, and the patient with the AIPL1 mutation developed microphthalmia and severe widespread retinal pigment epithelial atrophy, in contrast to previously reported cases. It is currently evident that mutation screening needs to be done in at least 18 genes known to be associated with LCA. Thus, homozygosity mapping is an alternative technique to improve the molecular diagnosis in LCA, which is a group of genetically and clinically heterogeneous diseases causing retinal degeneration. The patients without mutation in known genes may further be analyzed by using next-generation sequencing.


Inhibition of survivin expression to induce the apoptosis of hepatocarcinoma cells by adenovirus-mediated siRNA.
In order to provide more efficient transduction of plasmid siRNA into target cells, develop more susceptible transduction into cancer cell types, and more easily explore application in animal experiments, we examined development of an adenoviral vector-mediated siRNA expression system and inhibition of survivin gene expression to induce the growth and apoptosis of hepatocarcinoma cells. A system of adenoviral vector-mediated siRNA expression was constructed for the survivin gene. Survivin gene expression in HepG2 cells infected with recombinant adenovirus was detected by Western blot and RT-PCR, and apoptotic cells were investigated by FAC. Western blot analysis showed that the infection of adenovirus-mediated siRNA against survivin efficiently inhibited the expression of survivin in hepatocarcinoma cells with an inhibitory rate of 66.32%. Semi-quantitative RT-PCR showed that survivin gene mRNA transcription was reduced by nearly 72.34% with a peak at 72 h. The number of apoptotic cells increased. In conclusion, results demonstrated that this adenovirus-mediated siRNA system could serve as a useful tool for both basic research on the analysis of gene function and cancer therapy applications.


Toxicological Comparison of Pesticide Active Substances Approved for Conventional vs. Organic Agriculture in Europe.
There is much debate about whether the (mostly synthetic) pesticide active substances (AS) in conventional agriculture have different non-target effects than the natural AS in organic agriculture. We evaluated the official EU pesticide database to compare 256 AS that may only be used on conventional farmland with 134 AS that are permitted on organic farmland. As a benchmark, we used (i) the hazard classifications of the Globally Harmonized System (GHS), and (ii) the dietary and occupational health-based guidance values, which were established in the authorization procedure. Our comparison showed that 55% of the AS used only in conventional agriculture contained health or environmental hazard statements, but only 3% did of the AS authorized for organic agriculture. Warnings about possible harm to the unborn child, suspected carcinogenicity, or acute lethal effects were found in 16% of the AS used in conventional agriculture, but none were found in organic agriculture. Furthermore, the establishment of health-based guidance values for dietary and non-dietary exposures were relevant by the European authorities for 93% of conventional AS, but only for 7% of organic AS. We, therefore, encourage policies and strategies to reduce the use and risk of pesticides, and to strengthen organic farming in order to protect biodiversity and maintain food security.


[A new transplant for iridoplasty].
A new transplant is proposed for replacing extensive defects of the iris: cadaveric aortic wall. Experiments demonstrated that taking in of this graft courses as incomplete regeneration with partial taking in and replacement of the graft by cell elements of the iris with the formation of a cicatrice. Aortic wall transplant was used in the treatment of 4 patients.


Pilot study of Internet-based early intervention for combat-related mental distress.
This article evaluates an Internet-based early intervention combining online cognitive-behavioral therapy (CBT) with electronic peer-to-peer support intended to promote mental health and well-being among combat veterans. We conducted a phase 1 clinical trial of 50 Iraq and Afghanistan veterans using a pre and post single-arm design. We evaluated feasibility and changes in mental health symptoms (depression and posttraumatic stress disorder [PTSD]), functional status, and attitudes toward treatment seeking at baseline and weeks 4, 8, and 12. A diverse group of veterans was enrolled (26% ethnic minority, 90% male, 66% with income <$30,000/year, 88% with no prior treatment for depression). Participants completed a mean of 4 of 6 lessons (standard deviation = 2.54). From baseline to week 12, there were significant declines in the Center for Epidemiologic Studies-Depression scale score (effect size [ES] = 0.41) and PTSD Checklist-Military version score (ES = 0.53). There were significant improvements in willingness to accept diagnosis (ES = 1.08) and perceived social norms and stigma regarding friends (ES = 1.51). Although lack of a control group is a limitation, the Internet-based program combining CBT-based coping skills training and peer-to-peer support demonstrated potential feasibility and evidenced benefit in symptom remediation for depression and PTSD.


Borehole measurement of the newtonian gravitational constant.
It has been reported that the geophysically determined Newtonian gravitational constant is consistently larger than the laboratory value by 1 to 2% on the basis of gravity measurements in Australian mines. This discrepancy may have strong implications for the physics of gravitation. To test whether similar results can be observed in a different geological environment, gravity measurements in a Michigan borehole have been examined. Although these results cannot be taken as conclusive, owing to the large uncertainties involved in mass determination on a geophysical scale, these measurements are generally consistent with those of the Australian experiment.


Individualized norms of optimal fetal growth: fetal growth potential.
To demonstrate that individualized optimal fetal growth norms, accounting for physiologic and pathologic determinants of fetal growth, better identify normal and abnormal outcomes of pregnancy than existing methods. In a prospective cohort of 38,033 singleton pregnancies, we identified 9,818 women with a completely normal outcome of pregnancy and characterized the physiologic factors affecting birth weight using multivariable regression. We used those physiologic factors to individually predict optimal growth trajectory and its variation, growth potential, for each fetus in the entire cohort. By comparing actual birth weight with growth potential, population, ultrasound, and customized norms, we calculated for each fetus achieved percentiles, by each norm. We then compared proportions of pregnancies classified as normally grown, between 10th and 90th percentile, or aberrantly grown, outside this interval, by growth potential and traditional norms, in 14,229 complicated pregnancies, 1,518 pregnancies with diabetes or hypertensive disorders, and 1,347 pregnancies with neonatal complications. Nineteen physiologic factors, associated with maternal characteristics and early placental function, were identified. Growth potential norms correctly classified significantly more pregnancies than population, ultrasound, or customized norms in complicated pregnancies (26.4% compared with 18.3%, 18.7%, 22.8%, respectively, all P<.05), pregnancies with diabetes or hypertensive disorders (37.3% compared with 23.0%, 28.0%, 34.0%, respectively, all P<.05) and neonatal complications (33.3% compared with 19.7%, 24.9%, 29.8%, respectively, all P<.05). Growth potential norms based on the physiologic determinants of birth weight are a better discriminator of aberrations of fetal growth than traditional norms. II.


Chronic immune colitis in rabbits.
A chronic colitis has been induced in rabbits having many of the histological features of human ulcerative colitis. Animals were first immunised with the common enterobacterial antigen of Kunin and haemagglutinating antibodies demonstrated in high titre. An immune complex colitis was then established by the injection of soluble immune complexes following mild irritation of the rectum with dilute formalin as previously described. The rabbits developed an acute colitis within the first week but, in contrast with unsensitised rabbits, the inflammation persisted and was still present at six months as assessed by proctoscopy and rectal biopsy. Kunin-sensitised rabbits receiving intravenous saline, antigen, or antibody alone did not develop a chronic colitis. It is suggested that hypersensitivity to colonic bacterial antigens may be one mechanism whereby an acute colitis becomes chronic.


Beneficial effects of probiotics in upper respiratory tract infections and their mechanical actions to antagonize pathogens.
Probiotics are live micro-organisms with beneficial effects on human health, which have the ability to counteract infections at different locations of the body. Clinical trials have shown that probiotics can be used as preventive and therapeutic agents in upper respiratory tract infections (URTIs) and otitis. Their mechanical properties allow them to aggregate and to compete with pathogens for nutrients, space and attachment to host cells. Consequently, they can directly antagonize pathogens and thus exert beneficial effects without directly affecting the metabolism of the host. An overview of the probiotics with such traits, tested up to date in clinical trials for the prevention or treatment of URTIs and otitis, is presented in this review. Their mechanical properties in the respiratory tract as well as at other locations are also cited. Species with interesting in vitro properties towards pharyngeal cells or against common respiratory pathogens have also been included. The potential safety risks of the cited species are then discussed. This review could be of help in the screening of probiotic strains with specific mechanical properties susceptible to have positive effects in clinical trials against URTIs.


CD80 Plays a Critical Role in Increased Inflammatory Responses in Herpes Simplex Virus 1-Infected Mouse Corneas.
We recently reported that herpes simplex virus 1 (HSV-1) infection suppresses CD80 but not CD86 expression in vitro and in vivo This suppression required the HSV-1 ICP22 gene. We also reported that overexpression of CD80 by HSV-1 exacerbated corneal scarring in BALB/c mice. We now show that this recombinant virus (HSV-CD80) expressed high levels of CD80 both in vitro in cultured rabbit skin cells and in vivo in infected mouse corneas. CD80 protein was detected on the surface of infected cells. The virulence of the recombinant HSV-CD80 virus was similar to that of the parental strain, and the replication of HSV-CD80 was similar to that of control virus in vitro and in vivo Transcriptome analysis detected 75 known HSV-1 genes in the corneas of mice infected with HSV-CD80 or parental virus on day 4 postinfection. Except for significantly higher CD80 expression in HSV-CD80-infected mice, levels of HSV-1 gene expression were similar in corneas from HSV-CD80-infected and parental virus-infected mice. The number of CD8+ T cells was higher, and the number of CD4+ T cells was lower, in the corneas of HSV-CD80-infected mice than in mice infected with parental virus. HSV-CD80-infected mice displayed a transient increase in dendritic cells. Transcriptome analysis revealed mild differences in dendritic cell maturation and interleukin-1 signaling pathways and increased expression of interferon-induced protein with tetratricopeptide repeats 2 (Ifit2). Together, these results suggest that increased CD80 levels promote increased CD8+ T cells, leading to exacerbated eye disease in HSV-1-infected mice.IMPORTANCE HSV-1 ocular infections are the leading cause of corneal blindness. Eye disease is the result of a prolonged immune response to the replicating virus. HSV-1, on the other hand, has evolved several mechanisms to evade clearance by the host immune system. We describe a novel mechanism of HSV-1 immune evasion via ICP22-dependent downregulation of the host T cell costimulatory molecule CD80. However, the exact role of CD80 in HSV-1 immune pathology is not clear. In this study, we show that eye disease is independent of the level of HSV-1 replication and that viral expression of CD80 has a detrimental role in corneal scarring, likely by increasing CD8+ T cell recruitment and activation.


Prematurely born children demonstrate white matter microstructural differences at 12 years of age, relative to term control subjects: an investigation of group and gender effects.
The goal was to use diffusion tensor imaging to test the hypothesis that prematurely born children demonstrate long-term, white matter, microstructural differences, relative to term control subjects. Twenty-nine preterm subjects (birth weight: 600-1250 g) without neonatal brain injury and 22 matched, term, control subjects were evaluated at 12 years of age with MRI studies, including diffusion tensor imaging and volumetric imaging; voxel-based morphometric strategies were used to corroborate regional diffusion tensor imaging results. Subjects also underwent neurodevelopmental assessments. Neurodevelopmental assessments showed significant differences in full-scale, verbal, and performance IQ and Developmental Test of Visual Motor Integration scores between the preterm and term control subjects. Diffusion tensor imaging studies demonstrated widespread decreases in fractional anisotropy (a measure of fiber tract organization) in the preterm children, compared with the control subjects. Regions included both intrahemispheric association fibers subserving language skills, namely, the right inferior frontooccipital fasciculus and anterior portions of the uncinate fasciculi bilaterally, and the deep white matter regions to which they project, as well as the splenium of the corpus callosum. These changes in fractional anisotropy occurred in subjects with significant differences in frontal, temporal, parietal, and deep white matter volumes. Fractional anisotropy values in the left anterior uncinate correlated with verbal IQ, full-scale IQ, and Peabody Picture Vocabulary Test-Revised scores for preterm male subjects. In addition, preterm male subjects were found to have the lowest values for fractional anisotropy in the right anterior uncinate fasciculus, and fractional anisotropy values in that region correlated with both verbal IQ and Peabody Picture Vocabulary Test-Revised scores for the preterm groups; these findings were supported by changes identified with voxel-based morphometric analyses. Compared with term control subjects, prematurely born children with no neonatal ultrasound evidence of white matter injury manifest changes in neural connectivity at 12 years of age.


Penile prosthesis implantation in a transsexual neophallus.
Reconstruction surgery for a female to male transsexual usually involves mastectomy, hysterectomy and creating an aesthetically appealing neophallus. We have successfully inserted an inflatable prosthesis using the AMS CX prosthesis in a 45 year old transsexual, who had a large bulky neophallus constructed from the anterior abdominal subcutaneous fat, about 9 years ago. The single cylinder CX prosthesis was well anchored to the symphysis pubis using a dacron windsock tubing, the activation pump was placed in the dependent pouch of the right labium and the reservior in the usual perivesical space. The patient subsequently had debulking procedure using liposuction to create a more aesthetic and functional phallus. To date, the inflatable neophallus prosthesis is functioning well.


The potential for further development of passive safety.
In Europe, emphasis is being transferred from injury prevention to accident prevention to reduce road casualties. This study attempted to identify the current potential for serious casualty reduction using passive safety by examining the crash performance of new cars with seriously injured occupants. The Co-operative Crash Injury Study conducts in-depth investigations of around 1200 vehicles per year from seven sample regions around England. Attention was focussed on passenger cars manufactured from 2004 to 2008 with at least one occupant injured to AIS level 3 or more. 28% of MAIS 3+ occupants were unbelted and 40% were belted but involved in crashes with limited potential for passive protection. A further 32% of occupants were belted and involved in crashes with potential for improved crashworthiness design. For these occupants, five major functional requirements were identified for crashworthiness improvement: a reduction of seatbelt loads on the chest and abdomen in frontal crashes, particularly for seniors; reduction in femur and tibia loads in frontal crashes; provision of head and chest protection in near-side crashes; and reduction of occupant lateral excursion in far-side impacts. Together these functions accounted for 70% of the identified requirements. Other smaller requirements were identified, each contributing up to 5% of total. Overall, the case supporting further developments in passive safety still appears significant.


Effects of molybdate and tungstate on expression levels and biochemical characteristics of formate dehydrogenases produced by Desulfovibrio alaskensis NCIMB 13491.
Formate dehydrogenases (FDHs) are enzymes that catalyze the formate oxidation to carbon dioxide and that contain either Mo or W in a mononuclear form in the active site. In the present work, the influence of Mo and W salts on the production of FDH by Desulfovibrio alaskensis NCIMB 13491 was studied. Two different FDHs, one containing W (W-FDH) and a second incorporating either Mo or W (Mo/W-FDH), were purified. Both enzymes were isolated from cells grown in a medium supplemented with 1 μM molybdate, whereas only the W-FDH was purified from cells cultured in medium supplemented with 10 μM tungstate. We demonstrated that the genes encoding the Mo/W-FDH are strongly downregulated by W and slightly upregulated by Mo. Metal effects on the expression level of the genes encoding the W-FDH were less significant. Furthermore, the expression levels of the genes encoding proteins involved in molybdate and tungstate transport are downregulated under the experimental conditions evaluated in this work. The molecular and biochemical properties of these enzymes and the selective incorporation of either Mo or W are discussed.


The peptide nucleic acids: a new way for chromosomal investigation on isolated cells?
The development of nucleic acid analogues has become an important feature due to the potential use of this new biomolecular tool in genetic diagnostics and investigations. Among all the synthetic oligonucleotides designed, the peptide nucleic acids (PNA) constitute a remarkable class of nucleic acid mimics, with important physico-chemical properties which have been exploited to develop a wide range of powerful biomolecular tools, including molecular probes, biosensors and anti-gene agents. New applications of PNA involve their use as hybridization probes, and consequently the PNA technology is now developing within the field of in situ hybridization techniques. Recent studies have reported the successful use of centromeric PNA probes on human lymphocytes, sperm as well as on isolated oocytes and blastomeres. Muticolour PNA protocols have been described for the specific identification of several human chromosomes. These data show that PNA could become a powerful complement to fluorescence in situ hybridization (FISH) for in situ chromosomal investigation, especially on isolated cells. The present paper gives an overview of the properties of PNA and the assays exploiting PNA technology in molecular genetics and cytogenetics.


InSpire to Promote Lung Assessment in Youth: Evolving the Self-Management Paradigms of Young People With Asthma.
Asthma is the most common chronic disease in childhood, disproportionately affecting urban, minority, and disadvantaged children. Individualized care plans supported by daily lung-function monitoring can reduce morbidity and mortality. However, despite 20 years of interventions to increase adherence, only 50% of US youth accurately follow their care plans, which leads to millions of preventable hospitalizations, emergency room visits, and sick days every year. We present a feasibility study of a novel, user-centered approach to increasing young people's lung-function monitoring and asthma self-care. Promoting Lung Assessment in Youth (PLAY) helps young people become active managers of their asthma through the Web 2.0 principles of participation, cocreation, and information sharing. Specifically, PLAY combines an inexpensive, portable spirometer with the motivational power and convenience of mobile phones and virtual-community gaming. The objective of this study was to develop and pilot test InSpire, a fully functional interface between a handheld spirometer and an interactive game and individualized asthma-care instant-messaging system housed on a mobile phone. InSpire is an application for mobile smartphones that creates a compelling world in which youth collaborate with their physicians on managing their asthma. Drawing from design-theory on global timer mechanics and role playing, we incentivized completing spirometry maneuvers by making them an engaging part of a game young people would want to play. The data can be sent wirelessly to health specialists and return care recommendations to patients in real-time. By making it portable and similar to applications normally desired by the target demographic, InSpire is able to seamlessly incorporate asthma management into their lifestyle. We describe the development process of building and testing the InSpire prototype. To our knowledge, the prototype is a first-of-its kind mobile one-stop shop for asthma management. Feasibility testing in children aged 7 to 14 with asthma assessed likability of the graphical user interface as well as young people's interest in our incentivizing system. Nearly 100% of children surveyed said they would play games like those in PLAY if they involved breathing into a spirometer. Two-thirds said they would prefer PLAY over the spirometer alone, whereas 1/3 would prefer having both. No children said they would prefer the spirometer over PLAY. Previous efforts at home-monitoring of asthma in children have experienced rapid decline in adherence. An inexpensive monitoring technology combined with the computation, interactive communication, and display ability of a mobile phone is a promising approach to sustainable adherence to lung-function monitoring and care plans. An exciting game that redefines the way youth conduct health management by inviting them to collaborate in their health better can be an incentive and a catalyst for more far-reaching goals.


A coupled two-dimensional main chain torsional potential for protein dynamics: generation and implementation.
Main chain torsions of alanine dipeptide are parameterized into coupled 2-dimensional Fourier expansions based on quantum mechanical (QM) calculations at M06 2X/aug-cc-pvtz//HF/6-31G** level. Solvation effect is considered by employing polarizable continuum model. Utilization of the M06 2X functional leads to precise potential energy surface that is comparable to or even better than MP2 level, but with much less computational demand. Parameterization of the 2D expansions is against the full main chain torsion space instead of just a few low energy conformations. This procedure is similar to that for the development of AMBER03 force field, except unique weighting factor was assigned to all the grid points. To avoid inconsistency between quantum mechanical calculations and molecular modeling, the model peptide is further optimized at molecular mechanics level with main chain dihedral angles fixed before the calculation of the conformational energy on molecular mechanical level at each grid point, during which generalized Born model is employed. Difference in solvation models at quantum mechanics and molecular mechanics levels makes this parameterization procedure less straightforward. All force field parameters other than main chain torsions are taken from existing AMBER force field. With this new main chain torsion terms, we have studied the main chain dihedral distributions of ALA dipeptide and pentapeptide in aqueous solution. The results demonstrate that 2D main chain torsion is effective in delineating the energy variation associated with rotations along main chain dihedrals. This work is an implication for the necessity of more accurate description of main chain torsions in the future development of ab initio force field and it also raises a challenge to the development of quantum mechanical methods, especially the quantum mechanical solvation models.


Transcutaneous electrical nerve stimulation versus baclofen in spasticity: clinical and electrophysiologic comparison.
Clinical and electrophysiologic comparison of the efficacy of transcutaneous electrical nerve stimulation (TENS) and oral baclofen in the treatment of spasticity. Patients with spinal cord injury and spasticity were included in the study. Ten patients were assigned to oral baclofen and 11 to TENS groups. For the comparison of H-reflex variables, 20 healthy individuals were allocated to a control group. TENS was applied to the tibial nerve for 15 days at a frequency of 100 Hz. Clinical (spasm frequency scale, painful spasm scale, lower limb Ashworth score, clonus score, deep tendon reflex score, plantar stimulation response score) and electrophysiologic evaluations (H-reflex response at the highest amplitude, latency of maximum H-reflex, and ratio of H-reflex response at the highest amplitude to M response at maximum amplitude) of the lower limb and functional evaluations (functional disability score and FIM) were carried out in baclofen and TENS groups before and after treatment. Posttreatment evaluation was made 24 hrs after the 15th session in the TENS group. In addition, clinical spasticity scores and electrophysiologic variables were measured 15 mins after the first application and 15 mins after the 15th session. Significant improvement was detected in lower limb Ashworth score, spasm frequency scale, deep tendon reflex score, functional disability score, and FIM in the baclofen (P = 0.011, P = 0.014, P = 0.025, P = 0.004, and P = 0.005, respectively) and TENS (P = 0.020, P = 0.014, P = 0.025, P = 0.003, and P = 0.003, respectively) group after treatment. Decrease in H-reflex maximum amplitude was significant in the TENS group (P = 0.026). Most marked improvement was observed in the third evaluation, 15 mins after the 15th session, particularly in lower limb Ashworth score (P = 0.006) and H-reflex maximum amplitude (P = 0.006) in the TENS group. The percentage change in clinical, electrophysiologic, and functional variables caused by baclofen was not different from that caused by repeated applications of TENS in the short- and long-term evaluations (P > 0.05). TENS may be recommended as a supplement to medical treatment in the management of spasticity.


A pilot study on the percutaneous absorption of microfine titanium dioxide from sunscreens.
Many Australians are being advised to apply microfine titanium dioxide sunscreen daily from the cradle to the grave. However, there is a surprising lack of data on the percutaneous absorption of microfine titanium dioxide. A prospective pilot study was conducted to analyse the percutaneous absorption of microfine titanium dioxide from sunscreens. Selected patients scheduled to have skin surgery, applied titanium dioxide sunscreen to the skin daily for 2-6 weeks prior to their operation. After excision, the stratum corneum of the sample was stripped and the titanium concentration of the remaining epidermis and dermis was measured by inductively coupled plasma-mass spectrometry. The results from this pilot study showed that levels of titanium in the epidermis and dermis of subjects who applied microfine titanium dioxide to their skin were higher than the levels of titanium found in controls. Studies with larger cohorts are necessary to establish if this absorption is statistically significant.


DAF-16-dependent suppression of immunity during reproduction in Caenorhabditis elegans.
To further understand how the nematode Caenorhabditis elegans defends itself against pathogen attack, we analyzed enhanced pathogen resistance (epr) mutants obtained from a forward genetic screen. We also examined several well-characterized sterile mutants that exhibit an Epr phenotype. We found that sterility and pathogen resistance are highly correlated and that resistance in both epr and sterile mutants is dependent on DAF-16 activity. Our data indicate that a DAF-16-dependent signaling pathway distinct from previously described pathways is involved in the activation of genes that confer resistance to bacterial pathogens. The timing of DAF-16-dependent gene activation in sterile mutants coincides with the onset of embryonic development in wild-type animals, suggesting that signals from developing embryos normally downregulate the immune response.


Bidirectional tachycardia in a child. A study using His bundle electrography.
This is a report of a case of bidirectional tachycardia in a 6-year-old girl with no evidence of any structural abnormality of the heart. The patient had never received digitalis. The arrhythmia appeared to be precipitated by effort and emotional stress, and could be induced by increasing the heart rate by atrial pacing or isoprenaline administration. His bundle electrography showed that the arrhythmia was ventricular in origin. This emphasizes the importance of recording an effort electrocardiogram in all children with unexplained syncopal episodes, even when the resting electrocardiogram is normal.


Molecular forms and subunit structure of the acetylcholine receptor in the central nervous system of insects.
The nicotinic acetylcholine receptor as probed by alpha-bungarotoxin binding has been isolated from detergent-solubilized ganglionic membrane preparations from the insect, Locusta migratoria. The isolation and characterization of the receptor protein was achieved by preparation of membrane fragments, extraction by sodium deoxycholate, centrifugation on sucrose density gradient, affinity chromatography, gel electrophoresis, and immunoblotting. The purified receptor protein migrated as a single band on polyacrylamide when native (Mr = 250,000 to 300,000) but also under denaturing conditions (Mr = 65,000) and cross-reacted with some monoclonal antibodies against the Torpedo receptor. In immunohistochemical approaches using polyclonal antibodies the acetylcholine receptor antigenic sites could topochemically be identified at very distinct zones in the neuropil of locust ganglia. The results suggest that the acetylcholine receptor in the central nervous system of insects represents an oligomeric complex composed of four identical or very similar subunits and thus may represent a prototype of the recently proposed homo-oligomeric ancestral acetylcholine receptor.


Role for opinion leaders in promoting evidence-based surgery.
"Opinion leaders" can be identified by surgeons from among their peers, and opinion leaders have a role in promoting best surgical practice. Postal survey. Four hundred eighteen (77% response fraction) randomly selected fellows of the Royal Australasian College of Surgeons. Number of opinion leaders identified locally, statewide, and nationally; ratings of 22 possible attributes in conferring status as an opinion leader; and views about and ratings of the role of opinion leaders. Most respondents were unable to identify a local colleague whom they considered to be an opinion leader in their own specialty (mode, 0; and median, 1) or in surgery in general (mode, 0; and median, 0). Estimated numbers of opinion leaders were significantly higher at the state and national levels for the respondents' own specialty and for surgery in general (P<.001 for all). Surgical expertise and teaching skills were rated most highly as conferring status as an opinion leader. Academic and professional contributions received the lowest ratings. Most surgeons (88%; 95% confidence interval, 84%-91%) agreed that opinion leaders could influence them to change their practice. Opinion leaders were rated as "very influential" by significantly more surgeons than clinical audit (38% vs 27%, chi21 = 13.6, P<.001) and clinical practice guidelines (38% vs 24%, chi21 = 21.4, P<.001) (McNemar test for both). Australian surgeons support the concept of opinion leaders. Although few local colleagues whom they consider as fulfilling such a role can be identified, opinion leaders are evident at a national level. Once opinion leaders are identified using attributes ranked in our survey, interventional studies will further delineate their influence in improving evidence-based surgical practice.


Standard work-up of the low-risk kidney transplant candidate: a European expert survey of the ERA-EDTA Developing Education Science and Care for Renal Transplantation in European States Working Group.
Existing guidelines on the evaluation and preparation of recipients for kidney transplantation target the entire spectrum of patients with end-stage renal disease. Within the ERA-EDTA Developing Education Science and Care for Renal Transplantation in European States (DESCARTES) Working Group, it was proposed that in a subset of relatively young patients (<40 years) without significant comorbidities (such as diabetes or cardiovascular disease), the work-up for transplantation could be restricted to a small set of tests. Aiming for agreement between transplant centres across Europe, we surveyed the opinion of 80 transplant professionals from 11 European states on the composition of a minimal work-up. We show that there is a wide agreement among European experts that the work-up for kidney transplantation of the low-risk candidate, as opposed to the standard risk candidate, could include a limited number of investigations. However, there is some disagreement regarding the small number of diagnostic procedures, which is related to geographical location within Europe and the professional background of respondents. Based on the results of the survey, published guidelines and expert meetings by the DESCARTES Working Group, we have formulated a proposal for the work-up of low-risk kidney transplant candidates.


Characterization of volatile organic gunshot residues in fired handgun cartridges by headspace sorptive extraction.
In forensic investigation of firearm-related cases, determination of the residual amount of volatile compounds remaining inside a cartridge could be useful in estimating the time since its discharge. Published approaches are based on following the decrease of selected target compounds as a function of time by using solid phase micro-extraction (SPME). Naphthalene as well as an unidentified decomposition product of nitrocellulose (referred to as "TEA2") are usually employed for this purpose. However, reliability can be brought into question given their high volatility and low reproducibility of the extracted procedure. In order to identify alternatives and therefore develop improved dating methods, an extensive study on the composition and variability of volatile residues in nine different types of cartridges was carried out. Analysis was performed using headspace sorptive extraction (HSSE), which is a more exhaustive technique compared to SPME. One hundred sixty six compounds were identified (several of which for the first time), and it was observed that the final compositional characteristics of each residue were strongly dependent on its source. Variability of single identified compounds within and between different types of cartridge as well as their evolution over time were also studied. Many explosion products containing up to four aromatic rings were found to be globally present in high proportions. Twenty-seven of them (excluding naphthalene) also presented detectable decreases during the first 24 h. Therefore, they could be used as complementary target analytes in future dating methods.


Dietary quality 1 year after diagnosis of coronary heart disease.
The purpose of this ancillary study is to determine the quality of diets in patients with documented coronary heart disease (CHD). Dietary data were originally collected using a 24-hour dietary recall in 555 patients with CHD, 1 year after a diagnostic coronary angiography. Data used for this investigation were collected between March 2001 and November 2003. Patients were participants in a clinical trial to improve adherence to lipid-lowering medications. The Alternate Healthy Eating Index, an instrument designed to evaluate the degree to which a diet has the potential to prevent cardiovascular disease, measured dietary quality. Linear regression models were used to assess the association of dietary quality with patients' sociodemographic and clinical characteristics. Mean age of participants was 61 years, with an average body mass index of 30 (calculated as kg/m(2)). Sixty percent were men. Average daily caloric intake was 1,775 kcal, with 50% of calories derived from carbohydrates, 18% from protein, and 32% from total fat. Average Alternate Healthy Eating Index score was 30.8 out of a possible maximum score of 80. Only 12.4% of subjects met the recommended consumption of vegetables, 7.8% for fruit, 8% for cereal fiber, and 5.2% for trans-fat intake. Lower dietary quality was associated with lower total caloric intake, as well as with smoking, obesity, and lower educational level. A high proportion of patients reported poor dietary quality 1 year after experiencing a coronary event. Our data support continued efforts to enhance healthful dietary changes over time for secondary prevention of CHD. Dietary change should be emphasized with CHD patients who are less educated, smokers, or obese.